Novel lipid and lipid nanoparticle compositions for nucleic acid delivery

Abstract

The present application provides novel cationic lipid compounds applicable to lipid nanoparticles, which can target different tissues and organs for drug delivery. Furthermore, the lipid nanoparticles may further comprise at least one helper lipid. The novel cationic lipid compounds or lipid nanoparticle compositions containing the same can specifically deliver prophylactic / therapeutic agents, particularly nucleic acid components, to target organs.

Description

Technical Field

[0001] Cross-reference This application claims the priority of Chinese Patent Application No. 202211480456.3, filed on November 24, 2022, with the title "Novel Lipids and Lipid Nanoparticle Compositions for Nucleic Acid Delivery", which is hereby incorporated herein by reference in its entirety. Technical Field

[0002] The present disclosure generally relates to the field of molecular biology. More specifically, the present invention relates to the use of novel lipids and lipid nanoparticle compositions comprising the same in the organ-specific delivery of substances such as nucleic acids.

Background Art

[0003] Therapeutic or prophylactic nucleic acids have the potential to revolutionize vaccination, gene therapy, protein replacement therapy, and the treatment of other genetic diseases. Since the first clinical study on therapeutic nucleic acids was conducted in 2000, the research on the design and delivery methods of nucleic acid molecules has made great progress. However, nucleic acid pharmaceuticals (including therapeutic and prophylactic agents) still face several challenges, such as most liposome formulations accumulating through biological processes in the liver, thereby reducing the efficacy of the composition delivered to the target organ. Similarly, other therapeutic agents such as proteins and small molecule drugs can also benefit from organ-specific delivery. Various types of compounds such as chemical drugs exhibit significant cytotoxicity. When these compounds can be accurately delivered to the desired organ, fewer off-target effects and side effects are observed.

[0004] Currently, most lipid nanoparticle delivery systems passively target the liver. A common strategy for changing the target organ of lipid nanoparticles is to adjust the composition of the lipid nanoparticles. Several reports have shown that lipid nanoparticles composed of one or two types of lipids can achieve spleen targeting by adjusting the ratio of nucleic acid to nanoparticles (Stephan Grabbe et al., Translating Nanoparticulate-Personalized Cancer Vaccines into Clinical Applications: Case Study with RNA-Lipoplexes for the Treatment of Melanoma, 2016). However, this strategy requires improvements in terms of formulation stability, encapsulation efficiency, and other aspects. Furthermore, several reports (Cheng Qiang et al., Selective organ targeting (SORT) nanoparticles for tissue-specific mRNA delivery and CRISPR-Cas gene editing, 2020) have shown that adding a permanently anionic lipid to LNPs composed of four components—cationized lipids, steroids, phospholipids, and PEG-lipids—can also specifically target the spleen, but at the expense of increased composition and increased complexity in the formulation process. Therefore, lipid nanoparticle delivery systems with high delivery capacity, high stability, and low complexity continue to be developed. Summary of the Invention

[0005] The present application aims to use cationic lipid compounds in a nucleic acid drug delivery system, which can deliver nucleic acid drug molecules by targeting them to various tissues and organs, achieve efficient expression of nucleic acids, and reduce toxicity by keeping the expression level of nucleic acids low in the liver.

[0006] The present application provides novel cationic lipid compounds applicable to lipid nanoparticles, which can be targeted to different tissues and organs for drug delivery, wherein the cationic lipid compound is one or more of the compounds represented by formula (I) or pharmaceutically acceptable salts, prodrugs, stereoisomers or deuterated derivatives thereof:

[0007] [ka] In the formula, X1 and X2 are each independently -OC(=O)-, -C(=O)O-, -NHC(=O)-, or -C(=O)NH-; Y1 and Y2 are each independently -OC(=O)-, -C(=O)O-, -NR5C(=O)-, -C(=O)NR5- bond, optionally substituted C1-C8 alkylene, or optionally substituted C2-C8 alkenylene, where R5 is selected from hydrogen or straight or branched chain C1-C8 hydrocarbyl; L1 and L2 are each independently a bond or an optionally substituted C1-C10 alkylene; R3 and R4 each independently represent methyl, ethyl, propyl, hydroxymethyl, hydroxyethyl, hydroxypropyl, hydroxybutyl, [ka] or R3 and R4 together with the nitrogen attached thereto form a cyclic moiety, wherein R3 or R4 is selected from the group including hydrogen or deuterium, and the cyclic moiety is a 4- to 8-membered heterocycloalkyl selected from azetidin-1-yl, pyrrolidinyl, piperidin-1-yl, 4-hydroxypiperidin-1-yl, azepan-1-yl, morpholinyl, and 4-acetylpiperazin-1-yl; R1 and R2 are each independently H or deuterium or a straight or branched chain C1-C8 hydrocarbyl; m is an integer from 1 to 13, for example, m is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, or 13; n is an integer from 1 to 13, for example, n is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, or 13.

[0008] According to some specific and preferred embodiments, the cationic lipid compound is one or more compounds represented by the following structural formula:

[0009] [Table 1] TIFF2025538636000005.tif201149 TIFF2025538636000006.tif201149 TIFF2025538636000007.tif205149 TIFF2025538636000008.tif200149 TIFF2025538636000009.tif216149 TIFF2025538636000010.tif216149 TIFF2025538636000011.tif216149 TIFF2025538636000012.tif217149 TIFF2025538636000013.tif196149 TIFF2025538636000014.tif222149 TIFF2025538636000015.tif196149 TIFF2025538636000016.tif210149 TIFF2025538636000017.tif216149 TIFF2025538636000018.tif204149 TIFF2025538636000019.tif215149 TIFF2025538636000020.tif205149

[0010] In particular, "pharmaceutically acceptable salts" refers to salts of the compounds of the present disclosure that are pharmaceutically acceptable as defined above and have the desired pharmacological activity. Such salts include acid addition salts formed with the following acids: inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and the like; 1,2-ethanedisulfonic acid, 2-hydroxyethanesulfonic acid, 2-naphthalenesulfonic acid, 3-phenylpropionic acid, 4,4'-methylenebis(3-hydroxy-2-ene-1-carboxylic acid), 4-methylbicyclo[2.2.2]oct-2-ene-1-carboxylic acid, acetic acid, aliphatic monocarboxylic and dicarboxylic acids, aliphatic sulfuric acids, aromatic sulfuric acids, benzenesulfonic acid, benzoic acid, camphorsulfonic acid, carbonic acid, cinnamic acid, chlorosulfonic ... Organic acids such as enoic acid, cyclopentanepropionic acid, ethanesulfonic acid, fumaric acid, glucoheptonic acid, gluconic acid, glutamic acid, glycolic acid, heptanoic acid, hexanoic acid, hydroxynaphthoic acid, lactic acid, lauryl sulfuric acid, maleic acid, malic acid, malonic acid, mandelic acid, methanesulfonic acid, muconic acid, o-(4-hydroxybenzoyl)benzoic acid, oxalic acid, p-chlorobenzenesulfonic acid, phenyl-substituted alkanoic acids, propionic acid, p-toluenesulfonic acid, pyruvic acid, salicylic acid, stearic acid, succinic acid, tartaric acid, tert-butylacetic acid, and trimethylacetic acid. Pharmaceutically acceptable salts also include base addition salts, which may be formed when acidic protons present can react with inorganic or organic bases. Acceptable inorganic bases include, but are not limited to, sodium hydroxide, sodium carbonate, potassium hydroxide, aluminum hydroxide, and calcium hydroxide. Acceptable organic bases include, but are not limited to, ethanolamine, diethanolamine, triethanolamine, tromethamine, N-methylglucamine, etc. It should be recognized that the particular anion or cation comprising any salt of the present application is not critical, so long as the salt as a whole is pharmacologically acceptable.

[0011] In particular, a "prodrug" refers to a compound, such as a therapeutic agent, that can be converted into a biologically active compound under physiological conditions or upon dissolution. Prodrugs are generally rapidly transformed in vivo to produce the parent compound, for example, by hydrolysis in blood. Prodrug compounds generally have advantages of solubility, tissue compatibility, or delayed release in mammalian organisms. The term "prodrug" is also meant to include any covalently bonded carrier that releases the active compound in vivo upon administration to a mammalian subject. Prodrugs include compounds with a hydroxyl, amino, or sulfhydryl bonded to any group that is cleaved to form a free hydroxyl, free amino, or free sulfhydryl when the prodrug is administered to a mammalian subject. Examples of prodrugs include, but are not limited to, acetate, formate, or benzoate derivatives, or amide derivatives of amine functional groups.

[0012] In particular, "stereoisomers" or "optical isomers" are isomers of a particular compound that have the same atoms bonded to the same other atoms but differ in the three-dimensional arrangement of those atoms. "Enantiomers" are stereoisomers of a particular compound that are mirror images of each other, like left and right hands. "Diastereomers" are stereoisomers of a particular compound that are not enantiomers. Chiral molecules contain chiral centers (also called stereogenic centers or stereogenic centers), which can be any point (not necessarily an atom) within the molecule and contain multiple groups, and the interchange of any two groups produces a stereoisomer. In organic compounds, chiral centers are usually carbon, phosphorus, or sulfur atoms, although other atoms can also be stereogenic in organic and inorganic compounds. A molecule can have multiple stereocenters, giving rise to many of its stereoisomers. In compounds whose stereoisomerism is due to tetrahedral stereocenters (e.g., tetrahedral carbons), the total number of possible stereoisomers is not expected to exceed 2n, where n is the number of tetrahedral stereocenters. In molecules with symmetry, the number of stereoisomers is often less than the maximum possible number of stereoisomers. A 50:50 mixture of enantiomers is called a racemic mixture. Alternatively, a mixture of enantiomers may be enriched so that one enantiomer is present in greater than 50%. In general, enantiomers and / or diastereomers can be resolved or separated using techniques known in the art. For any stereocenter or chiral axis with undefined stereochemistry, it is understood that the stereocenter or chiral axis may exist as the R-, S-, or a mixture of the R- and S-forms (including racemic and non-racemic mixtures). As used herein, the phrase "substantially free of other stereoisomers" means that the composition contains 15% or less of the other stereoisomer or stereoisomers, more preferably 10% or less, even more preferably 5% or less, or most preferably 1% or less.

[0013] deuterium( 2 H or D) is a stable, non-radioactive isotope of hydrogen, approximately twice the mass of hydrogen (H), and is the most common hydrogen isotope.

[0014] The novel cationic lipids provided by the present application are guided targeted lipid delivery (GOLD) lipids that also have targeted delivery functionality.

[0015] The present application provides lipid nanoparticles comprising the above-described cationic lipid. The cationic lipid compound is one or more of the compounds represented by general formula (I) or pharmaceutically acceptable salts, prodrugs, or stereoisomers thereof. Furthermore, the lipid nanoparticles may further comprise at least one helper lipid, which may be mixed with a drug or pharmacologically active molecule / drug to achieve encapsulation and specific delivery to specific tissues and organs. Tissue and organ specificity is achieved by the cationic lipid within the lipid nanoparticle, i.e., guided targeted lipid delivery (GOLD) lipid. The helper lipid in the lipid nanoparticle is optionally one or more of a phospholipid, a steroid, a polymer-conjugated lipid, and a modifiable lipid. Preferably, the phospholipid is selected from any one of DOPE, DSPC, DPPC, DMPC, DOPC, POPC, and SM, or a combination thereof. Preferably, the steroid is selected from one or more of cholesterol, sitosterol, stigmasterol, and ergosterol, and more preferably, the steroid is cholesterol and sitosterol. Preferably, the polymer in the polymer-conjugated lipid is a polymeric compound formed by the covalent bonding of one or more small molecule repeating units, and the polymer is selected from polyethylene glycol, polylactic acid, polyamide, cationic polymer, polysarcosine (pSar), polylactic-co-glycolic acid (PLGA), polyamino acid, polypeptide, polypeptoid, etc. Preferably, the polymer-conjugated lipid is selected from polyethylene glycol-conjugated lipid, and further, the polyethylene glycol-conjugated lipid is selected from one or more of ALC-0159, PEG1000-DMG, PEG5000-DMG, PEG2000-DMG, and PEG2000-DSPE. Preferably, the modifiable lipid includes lipids modified with small molecule compounds, vitamins, carbohydrates, peptides, proteins, nucleic acids, lipopolysaccharides, inorganic molecules or particles, metal ions or particles, and combinations of the above substances. In the lipid nanoparticles, the molar ratio of cationic lipid (lipid for delivery to the target) to helper lipid is 1:(0.5 to 2), preferably 1:(0.6 to 1.5), and more preferably 1:(0.8 to 1.2).

[0016] The lipid nanoparticles can deliver therapeutic / prophylactic agents to target organs such as the lung, heart, brain, spleen, lymph nodes, bone, skeletal muscle, stomach, small intestine, large intestine / colorectum, kidney, bladder, breast, testes, ovaries, uterus, spleen, thymus, brainstem, cerebellum, spinal cord, eye, ear, tongue, or skin. Preferably, the target organ is the spleen.

[0017] In another aspect, the present application provides a composition comprising a therapeutic or prophylactic agent and the lipid nanoparticles described above. Preferably, the mass ratio of the lipid nanoparticles to the prophylactic or therapeutic agent in the composition is 10:1 to 100:1, preferably 20:1 to 50:1, and more preferably 20:1 to 30:1.

[0018] The composition has an average particle size of 90 nm to 600 nm, preferably 200 nm to 400 nm, and more preferably 200 nm to 300 nm.

[0019] The composition has a polydispersity index (PDI) of 0.001 to 0.5, preferably 0.001 to 0.45, and more preferably 0.001 to 0.4.

[0020] In a preferred embodiment, the therapeutic / prophylactic agent is a nucleic acid. Nucleic acids include any form of nucleic acid molecule, including, but not limited to, single-stranded DNA, double-stranded DNA, single-stranded RNA, double-stranded RNA, short isomers, plasmid DNA, complementary DNA / cDNA, antisense oligonucleotides / ASO, small interfering nucleic acids / siRNA, small activating nucleic acids / saRNA, asymmetric interfering nucleic acids / aiRNA, micronucleic acids / miRNA, miRNA agonists / antagonists (agomils / antagomils), dicer substrate nucleic acids, small hairpin nucleic acids / shRNA, transfer RNA (tRNA), messenger RNA / mRNA, circular RNA / circRNA, self-amplifying mRNA / samRNA, aptamers, and other forms of nucleic acid molecules known in the art or that may be discovered / prepared in the future.

[0021] The nucleic acid molecules may include naturally occurring nucleotides, or may include nucleotide mimetics or functional analogs, or may include chemically modified forms of nucleotides.

[0022] Functional nucleotide analogs include, but are not limited to, locked nucleic acids (LNA), peptide nucleic acids (PNA), and morpholine ring oligonucleotide nucleic acid mimetics or functional analogs, either alone or in combination.

[0023] The chemical modification of nucleotides can be located on the backbone bond of nucleic acid molecules. The backbone bond can be modified by replacing one or more oxygen atoms. The modification of backbone bond can include replacing at least one phosphodiester bond with a phosphorothioate bond.

[0024] The chemical modification of the nucleotide may be located on the nucleoside. The modification on the nucleoside may be located on the sugar or base of the nucleoside. The sugar modification on the nucleoside may be selected from one or more of 2'-fluororibose, ribose, 2'-deoxyribose, arabinose, and hexose.

[0025] The chemical modifications of nucleotides include 5-methylcytosine, pseudouridine, 1-methylpseudouridine, pyridin-4-one ribonucleoside, 5-azauridine, 2-thio-5-azauridine, 2-thiouridine, 4-thio-pseudouridine, 2-thio-pseudouridine, 5-hydroxyuridine, 3-methyluridine, 5-carboxymethyluridine, 1-carboxymethyl-pseudouridine, 5-propynyluridine, 1-propynyl-pseudouridine, 5-taurinomethyluridine, 1-taurinomethyl-pseudouridine, 5 -Taurinomethyl-2-thiouridine, 1-taurinomethyl-4-thiouridine, 5-methyluridine, 1-methyl-pseudouridine, 4-thio-1-methyl-pseudouridine, 2-thio-1-methyl-pseudouridine, 1-methyl-1-deaza-pseudouridine, 2-thio-1-methyl-1-deaza-pseudouridine, dihydrouridine, dihydropseudouridine, 2-thio-dihydrouridine, 2-thio-dihydropseudouridine, 2-methoxyuridine, 2-methoxy-4-thiouridine, 4-methoxypseudouridine, 4-methyl Pseudoisocytidine, 2-thio-cytidine, 2-thio-5-methyl-cytidine, 4-thio-pseudoisocytidine, 4-thio-1-methyl-pseudoisocytidine, 4-thio-1-methyl-1-deaza-pseudoisocytidine, ... Socytidine, Zebularine, 5-Aza-Zebularine, 5-Methyl-Zebularine, 5-Aza-2-Thio-Zebularine, 2-Thio-Zebularine, 2-Methoxy-Cytidine, 2-Methoxy-5-Methyl-Cytidine, 4-Methoxy-Pseudoisocytidine, 4-Methoxy-1-Methyl-Pseudoisocytidine, 2-Aminopurine, 2,6-Diaminopurine, 7-Deaza-Adenine, 7-Deaza-8-Aza-Adenine, 7-Deaza-2-Aminopurine, 7-Deaza-8-Aza-2-Aminopurine, 7-Deaza-2,6-Diaminopurine, 7-Deaza-8-Aza-2,6-Diaminopurine, 1-methyladenosine, N6-methyladenosine, N6-isopentenyladenosine, N6-(cis-hydroxyisopentenyl)adenosine, 2-methylthio-N6-(cis-hydroxyisopentenyl)adenosine, N6-glycinylcarbamoyladenosine, N6-threonylcarbamoyladenosine, 2-methylthio-N6-threonylcarbamoyladenosine, N6,N6-dimethyladenosine, 7-methyladenine, 2-methylthioadenine, 2-methoxyadenine, inosine, 1-methylinosine, wyosine, wyobutosine, 7-deaza The amino acid sequence may be selected from one or more of guanosine, 7-deaza-8-azaguanosine, 6-thio-guanosine, 6-thio-7-deazaguanosine, 6-thio-7-deaza-8-azaguanosine, 7-methylguanosine, 6-thio-7-methylguanosine, 7-methylinosine, 6-methoxyguanosine, 1-methylguanosine, N2-methylguanosine, N2,N2-dimethylguanosine, 8-oxoguanosine, 7-methyl-8-oxoguanosine, 1-methyl-6-thio-guanosine, N2-methyl-6-thio-guanosine, and N2,N2-dimethyl-6-thio-guanosine. These modifications may be random or site-specific.

[0026] In a preferred example, the nucleic acid is an mRNA capable of encoding at least one antigen or a fragment thereof or an epitope thereof, or capable of encoding a specific therapeutic protein, wherein the antigen is selected from a pathogenic antigen such as a tumor-associated antigen or a pathogenic microbial antigen. The mRNA may be monocistronic or polycistronic.

[0027] The present application further provides the use of the composition in the manufacture of a medicament.

[0028] The present application further provides a pharmaceutical product comprising the above-described composition and a pharmaceutically acceptable auxiliary substance. A pharmaceutical auxiliary substance is an excipient or additive used in the manufacture of a pharmaceutical product or formulation. It is a substance other than an active ingredient, whose safety has been reasonably evaluated and which is included in a pharmaceutical product. Pharmaceutical auxiliary substances form the formulation, function as a carrier, and enhance stability, as well as perform other important functions such as solubilization, dissolution assistance, and sustained or controlled release. They are important components that may affect the quality, safety, and efficacy of a pharmaceutical product. Pharmaceutical auxiliary substances are classified according to their effects and purposes into solvents, propellants, solubilizers, co-solvents, emulsifiers, colorants, binders, disintegrants, fillers, lubricants, wetting agents, osmotic pressure adjusters, stabilizers, glidants, flavoring agents, preservatives, suspending agents, coating agents, fragrances, anti-adherents, integrating agents, penetration enhancers, pH adjusters, buffers, plasticizers, surfactants, foaming agents, anti-foaming agents, thickeners, encapsulating agents, humectants, absorbents, diluents, flocculating agents, deflocculants, filter aids, release retardants, etc.

[0029] Beneficial effects of this application: The present invention provides novel cationic lipids and lipid nanoparticles containing the same that can preferentially deliver therapeutic / prophylactic agents, particularly nucleic acid components, to specific organs, particularly organs other than the liver. This provides additional options for the delivery of nucleic acid drugs, gene vaccines, etc., and is particularly significant for the development and application of nucleic acid prophylactic / therapeutic agents.

[0030] Conventional lipid nanoparticles are generally delivered to the liver. However, the use of novel cationic lipid molecules of the present application can alter the targeting properties of lipid nanoparticles containing novel cationic lipid molecules. For lipid nanoparticles such as TMF2-TMF53 containing the novel cationic lipids of the present application, the ratio of nucleic acid drug distribution (fluorescence intensity) in the spleen to the liver is 1:70 (Table 5). However, conventional lipid nanoparticles without the addition of the present cationic lipids generally have higher liver delivery efficiency (Ansell, S.M., Du, X. Novel Lipids and Lipid Nanoparticle Formulations for Delivery of Nucleic Acids. WO2017075531 A1). [Brief explanation of the drawings]

[0031] [Figure 1] 1H NMR of compound 1. [Figure 2] 1H NMR of compound 2. [Figure 3] 1H NMR of compound 3. [Figure 4] 1H NMR of compound 4. [Figure 5] 1H NMR of compound 5. [Figure 6] 1H NMR of compound 6. [Figure 7] 1H NMR of compound 7. [Figure 8] 1H NMR of compound 8. [Figure 9] 1H NMR of compound 17. [Figure 10] 1H NMR of compound 18. [Figure 11] 1H NMR of compound 19. [Figure 12] 1H NMR of compound 20. [Figure 13] 1H NMR of compound 21. [Figure 14] 1H NMR of compound 22. [Figure 15] 1H NMR of compound 23. [Figure 16] 1H NMR of compound 24. [Figure 17] 1H NMR of compound 25. [Figure 18] 1H NMR of compound 27. [Figure 19] 1H NMR of compound 30. [Figure 20] 1H NMR of compound 33. [Figure 21] 1H NMR of compound 34. [Figure 22] 1H NMR of compound 35. [Figure 23]1H NMR of compound 36. [Figure 24] 1H NMR of compound 37. [Figure 25] 1H NMR of compound 38. [Figure 26] 1H NMR of compound 43. [Figure 27] 1H NMR of compound 44. [Figure 28] 1H NMR of compound 45. [Figure 29] 1H NMR of compound 46. [Figure 30] 1H NMR of compound 47. [Figure 31] 1H NMR of compound 48. [Figure 32] 1H NMR of compound 49. [Figure 33] 1H NMR of compound 50. [Figure 34] 1H NMR of compound 51. [Figure 35] 1H NMR of compound 52. [Figure 36] 1H NMR of compound 53. [Figure 37] 1H NMR of compound 54. [Figure 38] 1H NMR of compound 55. [Figure 39] 1H NMR of compound 56. [Figure 40] 1H NMR of compound 57. [Figure 41] 1H NMR of compound 58. [Figure 42] 1H NMR of compound 59. [Figure 43] 1H NMR of compound 60. [Figure 44] 1H NMR of compound 63. [Figure 45] 1H NMR of compound 64. [Figure 46] 1H NMR of compound 65. [Figure 47] 1H NMR of compound 66. [Figure 48] 1H NMR of compound 67. [Figure 49] 1H NMR of compound 68. [Figure 50] 1H NMR of compound 70. [Figure 51] 1H NMR of compound 76. [Figure 52] 1H NMR of compound 78. [Figure 53] 1H NMR of compound 79. [Figure 54] 1H NMR of compound 80. [Figure 55] 1H NMR of compound 81. [Figure 56] This graph shows the fluorescent signals in various organs of a mouse 4 hours after intravenous administration of a sample of lipid nanoparticle TMF1 loaded with luciferase mRNA. As shown in the graph, the fluorescent signal is strongest in the mouse liver, suggesting that the luciferase mRNA loaded on TMF1 is primarily expressed in the liver. [Figure 57] This graph shows the fluorescent signals in various organs of mice 4 hours after intravenous administration of a sample of lipid nanoparticles TMF10 loaded with luciferase mRNA. As shown in the graph, the fluorescent signal is strongest in the mouse spleen and very weak in the liver. The expression of luciferase mRNA loaded in TMF10 is shown to be significantly higher in the spleen than in the liver. [Figure 58] 1 shows summary data of the fluorescence intensity ratios of various lipid nanoparticles delivering luciferase mRNA to the spleen and liver. DETAILED DESCRIPTION OF THE INVENTION

[0032] A. Chemical and Formulation Definitions When used in the context of chemical groups, "hydrogen" refers to -H and "deuterium" refers to 2H or D, "hydroxyl" refers to -OH, "oxo" refers to =O, "carbonyl" refers to -C(=O)-, "carboxyl" refers to -C(=O)OH (also written as -COOH or -COH), "halo" independently refers to -F, -Cl, -Br, or -I, "amino" refers to -NH, "hydroxyamino" refers to -NHOH, "nitro" refers to -NO, imino refers to =NH, "cyano" refers to -CN, and "isocyanato" refers to -N=C=O; "Azide" refers to -N3; ​​in the context of monovalent groups, "phosphate" refers to -OP(O)(OH)2 or its deprotonated form; in the context of divalent groups, "phosphate" refers to -OP(O)(OH)O- or its deprotonated form; "sulfhydryl" refers to -SH; "thio" refers to =S; "sulfonyl" refers to -S(O)2-; "hydroxysulfonyl" refers to -S(O)2OH; "sulfonamide" refers to -S(O)2NH2; and "sulfinyl" refers to -S(O)-. In the context of chemical formulas, the symbol "-" refers to a single bond, "=" refers to a double bond, and "≡" refers to a triple bond. The symbol "----" represents an optional bond, which, if present, is a single or double bond. When drawn perpendicular to a bond, the symbol [ka] indicates the attachment point of the group. To ensure that the reader can clearly identify the attachment point, it should be noted that attachment points are generally identified in this way only for large groups. [ka] refers to a single bond, where the group attached to the tip of the wedge "comes out of the page." [ka] indicates a single bond, and the group attached to the tip of the wedge is "into the plane of the paper." [ka] refers to a single bond; the geometry around a double bond (e.g., E or Z) is undefined. Thus, both options and combinations thereof are contemplated. Any undefined valence on an atom of a structure shown in this application implicitly represents a hydrogen atom bonded to that atom. A bold dot on a carbon atom indicates that the hydrogen bonded to the carbon is pointing out of the plane of the paper.

[0033] The term "alkyl," used without the "substituted" modifier, refers to a monovalent saturated aliphatic group having a carbon atom as the point of attachment, having a straight or branched acyclic structure, and containing no atoms other than carbon and hydrogen. -CH(Me), -CHCH(Et), -CHCHCH(n-Pr or propyl), -CH(CH)(i-Pr, iPr or isopropyl), -CHCHCHCH(n-Bu), -CH(CH)CHCH(sec-butyl), -CHCH(CH)(isobutyl), -C(CH)(tert-butyl, t-butyl, t-Bu, or tBu), and -CHC(CH)(neopentyl) groups are non-limiting examples of alkyl. The term "dialkyl," used without the "substituted" modifier, refers to a divalent saturated aliphatic group having one or two saturated carbon atoms as points of attachment, a straight or branched acyclic structure, no carbon-carbon double or triple bonds, and no atoms other than carbon and hydrogen. The groups -CH-(methylene), -CHCH-, -CHC(CH)CH-, and -CHCHCH- are non-limiting examples of dialkyl. "Alkane" refers to the class of compounds having the formula HR, where R is alkyl as defined above. When any of these terms are used with the modifier "substituted," one or more hydrogen atoms are independently replaced with -OH, -F, -Cl, -Br, -I, -NH, -NO, -COH, -COCH, -CN, -SH, -OCH, -OCHCH, -C(O)CH, -NHCH, -NHCHCH, -N(CH), -C(O)NH, -C(O)NHCH, -C(O)N(CH), -OC(O)CH, -NHC(O)CH, -S(O)OH, or -S(O)NH. The following groups are non-limiting examples of substituted alkyls: -CHOH, -CHCl, -CF, -CHCN, -CHC(O)OH, -CHC(O)OCH, -CHC(O)NH, -CHC(O)CH, -CHOCH, -CHOC(O)CH, -CHNH, -CHN(CH), and -CHCHCl.The term "haloalkyl" is a subset of substituted alkyl where hydrogen atom replacement is limited to halo (i.e., -F, -Cl, -Br, or -I) and no atoms other than carbon, hydrogen, and halogen are present. The group -CH2Cl is a non-limiting example of haloalkyl. The term "fluoroalkyl" is a subset of substituted alkyl where hydrogen atom replacement is limited to fluoro and no atoms other than carbon, hydrogen, and fluorine are present. The groups -CH2F, -CF3, and -CH2CF3 are non-limiting examples of fluoroalkyl.

[0034] The term "alkenyl," used without the "substituted" modifier, refers to a monovalent unsaturated aliphatic group having a carbon atom as a point of attachment, a straight or branched acyclic structure, at least one non-aromatic carbon-carbon double bond, no carbon-carbon triple bond, and no atoms other than carbon and hydrogen. Non-limiting examples include -CH=CH2 (vinyl), -CH=CHCH3, -CH=CHCH2CH3, -CH2CH=CH2 (allyl), -CH2CH=CHCH3, and -CH=CHCH=CH2. The term "alkenediyl," used without the "substituted" modifier, refers to a divalent unsaturated aliphatic group having two carbon atoms as points of attachment, a straight or branched acyclic structure, a straight or branched acyclic structure, at least one non-aromatic carbon-carbon double bond, no carbon-carbon triple bond, and no atoms other than carbon and hydrogen. The groups -CH=CH-, -CH=C(CH3)CH2-, -CH=CHCH2-, and -CH2CH=CHCH2- are non-limiting examples of alkenediyl groups. It should be noted that although alkenediyl groups are aliphatic, the possibility of the group forming part of an aromatic structure when attached to both ends is not excluded. The terms "olefin" and "chain olefin" are synonymous and refer to the class of compounds having the formula HR, where R is alkenyl as defined above. Similarly, the terms "terminal olefin" and "α-olefin" are synonymous and refer to an olefin with only one carbon-carbon double bond that is part of the vinyl at the end of the molecule. When any of these terms are used with the modifier "substituted," one or more hydrogen atoms are independently replaced with -OH, -F, -Cl, -Br, -I, -NH, -NO, -COH, -COCH, -CN, -SH, -OCH, -OCHCH, -C(O)CH, -NHCH, -NHCHCH, -N(CH), -C(O)NH, -C(O)NHCH, -C(O)N(CH), -OC(O)CH, -NHC(O)CH, -S(O)OH, or -S(O)NH. The groups -CH=CHF, -CH=CHCl, and -CH=CHBr are non-limiting examples of substituted alkenyl.

[0035] The term "alkynyl," used without the "substituted" modifier, refers to a monovalent, unsaturated aliphatic group having a carbon atom as the point of attachment, a straight or branched acyclic structure, at least one carbon-carbon triple bond, and no atoms other than carbon and hydrogen. As used herein, the term alkynyl does not exclude the presence of one or more non-aromatic carbon-carbon double bonds. The groups -C≡CH, -C≡CCH, and -CHC≡CCH are non-limiting examples of alkynyl. "Alkyne" refers to the class of compounds having the formula HR where R is alkynyl. When any of these terms are used with the modifier "substituted," one or more hydrogen atoms are independently replaced with -OH, -F, -Cl, -Br, -I, -NH, -NO, -COH, -COCH, -CN, -SH, -OCH, -OCHCH, -C(O)CH, -NHCH, -NHCHCH, -N(CH), -C(O)NH, -C(O)NHCH, -C(O)N(CH), -OC(O)CH, -NHC(O)CH, -S(O)OH, or -S(O)NH.

[0036] "Prevention" or "preventing" includes (1) inhibiting the onset of a disease in a subject or patient who is at risk and / or may be susceptible to the disease, but who has not yet experienced or displayed some or all of the disease state or symptoms, and / or (2) delaying the onset of a disease state or symptom in a subject or patient who is at risk and / or may be susceptible to the disease, but who has not yet experienced or displayed some or all of the disease state or symptom.

[0037] "Treatment" or "treating" includes (1) inhibiting a disease (e.g., preventing further progression of the condition and / or symptoms) in a subject or patient experiencing or exhibiting a disease state or symptom, (2) ameliorating a disease (e.g., ameliorating the condition and / or symptoms) in a subject or patient experiencing or exhibiting a disease state or symptom, and / or (3) achieving measurable relief of a disease in a subject or patient experiencing or exhibiting a disease state or symptom.

[0038] "Protein," "polypeptide," or "peptide" refers to a polymer of amino acid residues, which includes a wide range of protein molecules, such as cytokines, chemokines, interleukins, interferons, growth factors, clotting factors, anticoagulants, blood factors, bone morphogenetic proteins, immunoglobulins, enzymes, etc. Non-limiting examples of therapeutic proteins include the following therapeutic proteins or fragments, variants, or derivatives thereof: acid sphingomyelinase, adipotide, agalsidase-β, alglucosidase, α-galactosidase A, α-glucosidase, α-L-iduronidase, α-N-acetylglucosaminidase, amphiregulin, angiopoietins (Ang1, Ang2, Ang3, Ang4, ANGPTL2, ANGPTL3, ANGPTL4, ANGPTL5, ANGPTL6, ANGPTL7, ANGPTL8, ANGPTL9, ANGPTL10, ANGPTL11, ANGPTL12, ANGPTL13, ANGPTL14, ANGPTL15, ANGPTL16, ANGPTL17, ANGPTL18, ANGPTL19, ANGPTL20, ANGPTL21, ANGPTL22, ANGPTL23, ANGPTL24, ANGPTL25, ANGPTL26, ANGPTL27, ANGPTL28, ANGPTL29 ...9, ANGPTL29, ANGPTL29, ANGPTL21, ANGPTL21, ANGP NGPTL6, ANGPTL7), β-cellulin, β-glucuronidase, bone morphogenetic proteins (BMPs (BMP1, BMP2, BMP3, BMP4, BMP5, BMP6, BMP7, BMP8a, BMP8b, BMP10, BMP15)), CLN6 protein, epidermal growth factor (EGF), epigen, epiregulin, fibroblast growth factor (FGF (FGF-1, FGF-2, FGF-3, FGF-4, FGF-5, FGF-6, FGF-7, FGF-8, FGF-9, FGF-10, FGF-15)), β-glucuronidase, ... FGF-11, FGF-12, FGF-13, FGF-14, FGF-16, FGF-17, FGF-18, FGF-19, FGF-20, FGF-21, FGF-22, FGF-23), galsulfase, ghrelin, glucocerebrosidase, GM-CSF, heparin-binding EGF-like growth factor (HB-EGF), hepatocyte growth factor (HGF), hepcidin, human albumin, increased albumin loss, idursulfase (iduronate-2-sulfatase), integrin αVβ3, αVβ5 and α5β1, iduronate sulfatase, laronidase, N-acetylgalactosamine-4-sulfatase (rhASB, galsulfase, arylsulfatase A (ARSA), arylsulfatase B (ARSB)), N-acetylglucosamine-6-sulfatase, nerve growth factor (NGF) (brain-derived neurotrophic factor (BDNF)), neurotrophin-3 (NT-3) and neurotrophin-4 / 5 (NT-4 / 5), neuregulin (NRG1,NRG2, NRG3, NRG4), neuropilin (NRP-1, NRP-2), obestatin, platelet-derived growth factor (PDGF (PDFF-A, PDGF-B, PDGF-C, PDGF-D)), TGFβ receptor (endoglin, TGF-β1 receptor, TGF-β2 receptor, TGF-β3 receptor), thrombopoietin (THPO) (megakaryocyte growth and development factor (MGDF)), transforming growth factor (TGF (TGF-a, TGF-β (TGFβ1, TGFβ2, and TGFβ3))), VEGF (VEGF-A, VEGF-B, VEGF-C, VEGF-D, VEGF-E, VEGF-F, PIGF), nesiritide, trypsin, adrenocorticotropic hormone (ACTH), atrial natriuretic peptide (ANP), cholecystokinin, gastrin, leptin, oxytocin, somatostatin, vasopressin (antidiuretic hormone), calcitonin, exenatide, growth hormone (GH), somatotropin, insulin, insulin-like growth factor 1 (IGF-1), mecasermin linfabate, IGF-1 analog, mecasermin, IGF-1 analog, pegvisomant, pramlintide, teriparatide (human parathyroid hormone Therapeutic proteins for the treatment of metabolic or endocrine disorders, including monocyte residues 1-34), becapremin, diboternin-α (bone morphogenetic protein 2), histrelin acetate (gonadotropin-releasing hormone, GnRH), octreotide, and palifermin (keratinocyte growth factor, KGF); alteplase (tissue plasminogen activator, tPA), anistreplase, antithrombin III (AT-III), bivalirudin, darbepoetin-α, drotrecodin-α (activated protein C), erythropoietin, epoetin-α, erythropoietin tin, hematopoietin, factor IX, factor VIIa, factor VIII, lepirudin, protein C concentrate, reteplase (deletion mutant protein of tPA), streptokinase, tenecteplase, urokinase, angiostatin, anti-CD22 immunotoxin, denileukin diftitox, immunocyanin, MPS (metallopastimulin), aflibercept, endostatin, collagenase, human deoxyribonuclease I, dornase, hyaluronidase, papain, L-asparaginase, PEG-asparaginase, rasburicase,Hematological disorders, cardiovascular disorders, respiratory disorders, cancer or neoplastic diseases, infectious diseases, or immune disorders, including human chorionic gonadotropin (HCG), human follicle-stimulating hormone (FSH), lutropin-α, prolactin, α-1-protease inhibitor, lactase, pancreatic enzymes (lipase, amylase, protease), adenosine deaminase (bovine pegademase, PEG-ADA), abatacept, alefacept, anakinra, etanercept, interleukin-1 (IL-1) receptor antagonists, anakinra, thymosin, TNF-α antagonists, enfuvirtide, and thymosin α-1 a therapeutic protein for treating allergies; a therapeutic protein selected from human adjuvant proteins, in particular adjuvant or immunostimulatory proteins including the pattern recognition receptors TLR1, TLR2, TLR3, TLR4, TLR5, TLR6, TLR7, TLR8, TLR9, TLR10, TLR11; NOD1, NOD2, NOD3, NOD4, NOD5, NALP1, NALP2, NALP3, NALP4, NALP5, NALP6, NALP6, NALP7, NALP7, NALP8, NALP9, NALP10, NALP11, NALP12, NALP P13, NALP14, IIPAF, NAIP, CIITA, RIG-I, MDA5, and LGP2, signal transduction factors of TLR signaling (including adaptor proteins such as Trif and Cardif); components of small GTPase signaling (RhoA, Ras, Rac1, Cdc42, Rab, etc.); components of PIP signaling (PI3K, Src kinase, etc.); components of MyD88-dependent signaling (MyD88, IRAK1, IRAK2, IRAK4, TIRAP, TRAF6, etc.); components of MyD88-independent signaling components (TICAM1, TICAM2, TRAF6, TBK1, IRF3, TAK1, IRAK1, etc.); activating kinases (e.g., Akt, MEKK1, MKK1, MKK3, MKK4, MKK6, MKK7, ERK1, ERK2, GSK3, PKC kinase, PKD kinase, GSK3 kinase, JNK, p38MAPK, TAK1, IKK, and TAK1); activating transcription factors (e.g., NF-κB, c-Fos, c-Jun, c-Myc, CREB, AP-1, Elk-1, ATF2, IRF-3, IRF-7), heat shock proteins (HSP10,HSP60, HSP65, HSP70, HSP75, HSP90), gp96, fibrinogen, fibronectin type III repeat additional domain A); or components of the complement system (C1q, MBL, C1r, C1s, C2b, Bb, D, MASP-1, MASP-2, C4b, C3b, C5a, C3a, C4a, C5b, C6, C7, C8, C9, CR1, CR2, CR3, CR4, C1qR, C1INH, C4bp, MCP, DAF, H, I, P, and CD59), or inducible target genes (e.g., β-defensins, cell surface proteins), or human adjuvant proteins including trif, flt-3 ligand, Gp96, or fibronectin, IL-1α, IL1β, IL-2, IL-6, IL-7, IL-8, IL-9, IL-12, IL-13, IL-15, IL-16, Cytokines that induce or enhance innate immune responses, including IL-17, IL-18, IL-21, IL-23, TNFα, IFNα, IFNβ, IFNγ, GM-CSF, G-CSF, and M-CSF; chemokines, including IL-8, IP-10, MCP-1, MIP-1α, RANTES, eotaxin, and CCL21; cytokines released by macrophages, such as IL-1, IL-6, IL-8, IL-12, and TNF-α; as well as IL-1R1 and IL-1α; and bacterial (adjuvant) proteins, especially bacterial heat shock proteins or chaperones (Hsp60, Hsp70, Hsp90, Hsp1). sp100); OmpA (outer membrane protein) from Gram-negative bacteria; bacterial porin bacterial toxins including OmpF, pertussis toxin (PT) from Bordetella pertussis, pertussis adenylate cyclase toxins CyaA and CyaC from Bordetella pertussis, PT-9K / 129G mutant from pertussis toxin, pertussis adenylate cyclase toxins CyaA and CyaC from Bordetella pertussis, tetanus toxin, cholera toxin (CT), cholera toxin B subunit, CTK63 mutant from cholera toxin, CTE112K mutant from CT, Escherichia coli heat-labile enterotoxin (LT), B subunit from heat-labile enterotoxin (LTB), LTK63,Virulence-reduced Escherichia coli heat-labile enterotoxin mutants containing LTR72; phenol-soluble modulin; neutrophil-activating protein (HP-NAP) from Helicobacter pylori; surfactant protein D; outer surface protein A lipoprotein from Borrelia burgdorferi; Ag38 (38 kDa antigen) from Mycobacterium tuberculosis; proteins derived from bacterial fimbriae; enterotoxin CT from Vibrio cholerae, pilin derived from gram-negative bacterial fimbriae, surfactant protein A and bacterial flagellin, protozoan (adjuvant) Adjuvant proteins, especially Tc52 from Trypanosoma cruzi, PFTG from Trypanosoma gondii, protozoan heat shock proteins, LeIF from Leishmania spp., profilin-like protein from Toxoplasma gondii, viral (adjuvant) proteins, especially respiratory syncytial virus fusion glycoprotein (F protein), envelope protein from MMT virus, murine leukemia virus proteins, wild-type measles virus hemagglutinin protein, fungal (adjuvant) proteins , particularly fungal immunomodulatory proteins (FIP, LZ-8); and keyhole limpet hemocyanin (KLH), OspA; therapeutic proteins for hormone replacement therapy, particularly estrogen, progesterone or progestin, and testosterone; therapeutic proteins for reprogramming somatic cells into pluripotent or totipotent stem cells, particularly Oct-3 / 4, the Sox gene family (Sox1, Sox2, Sox3, and Sox15), the Klf family (Klf1, Klf2, Klf4, and Klf5), the Myc family, therapeutic antibodies selected from antibodies for treating cancer or tumor diseases, in particular 131T-tositumomab, 3F8, 8H9, abagovomab, adecatumumab, afutuzumab, alacizumab pegol, alemtuzumab, amatuximab, AME-133v, AMG102, anatumomab mafenatox, apolizumab, bavituximab, bectumab, berinnumab, bevacizumab, bivatuzumab-DM1, blinatumomab, brentuximab vedotin, cantuzumab, cantuzumab mertansine, cantuzumab ravtansine, capromab pendetide, carlumab, catumaxomab, cetuximab,sitatuzumab bogatox, cixutumumab, clivatuzumab tetraxetan, CNTO328, CNTO 95, conatumumab, dacetuzumab, dalotuzumab, denosumab, detumomab, drozitumab, ecromeximab, edrecolomab, elotuzumab, ercilimab, enavatuzumab, ensituximab, epratuzumab, ertumaxomab, ertumaxomab, etaracizumab, farletuzumab, FBTA05, ficlatuzumab, figitumumab, framvotumab, galiximab, , Galiximab, Ganitumab, GC1008, Gemtuzumab, Gemtuzumab ozogamicin, Girentuximab, Glenbatumumab vedotin, GS6624, HuC242-DM4, HuHMFG1, HuN901-DM1, Ibritumomab, Icrucumab, ID09C3, Indatuximab ravtansine, Inotuzumab ozogamicin, Intetumumab, Ipilimumab, Iratumumab, Labetuzumab, Lexatumumab, Lintuzumab, Lorvotuzumab Mertansine, lucatumumab, rumiliximab, mapatumumab, matuzumab, MDX-060, MEDI522, mitumomab, mogamulizumab, MORab-003, MORab-009, moxetumomab pasudotox, MT103, nacolomab tafenatox, naptumomab estafenatox, narunatumumab, necitumumab, nimotuzumab, nimotuzumab, olaratumab, onartuzumab, oportuzumab Monatox, oregovomab, oregovomab, PAM4, panitumumab, patritumumab, pemtumomab, pertuzumab, priliximab, racotumomab, radletumumab, ramucirumab, rilotumumab, rituximab, lobatumumab, samalizumab, SGN-30, SGN-40, sibrotuzumab, siltuximab, tabalumab, tacatuzumab tetraxetan, taplitumumab paptox, tenatumomab, teprotumumab, TGN1412, ticilimumab (=tremelimumab), tigatuzumab, TNX-650, tositumomab, trastuzumab, TRBS07, tremelimumab, TRU-016, TRU-016, tucotuzumab Celmoleukin, ublituximab, urelumab, veltuzumab, veltuzumab (IMMU-106), volociximab, votumumab, WX-G250, zalutumumab, and natalizumab;Antibodies for treating immune disorders, in particular efalizumab, epratuzumab, etrolizumab, fontolizumab, ixekizumab, mepolizumab, milatuzumab, pooled immunoglobulins, priliximab, rituximab, rontalizumab, ruplizumab, sarilumab, vedolizumab, visilizumab, reslizumab, adalimumab, acelizumab, atinumab, atlizumab, belimumab, besilesomab, BMS-945429, briakinumab, brodalumab, canakinumab, and certolizumab Pegol, erlizumab, fezakinumab, golimumab, gomiliximab, infliximab, mavrilimumab, natalizumab, ocrelizumab, odulimab, ofatumumab, ozoralizumab, pexelizumab, rovelizumab, SBI-087, SBI-087, secukinumab, sirukumab, talizumab, tocilizumab, toralizumab, TRU-015, TRU-016, ustekinumab, ustekinumab, beparizumab molumab, zolimomab alitox, sifalimumab, lumiliximab, and Rho(D) immunoglobulin; antibodies for the treatment of infectious diseases, in particular afelimab, CR6261, edovacomab, efungumab, exvivirumab, felvizumab, foravirumab, ibalizumab, ribivirumab, motavizumab, nebacumab, tuvilumab, urtoxazumab, bavituximab, pagibaximab, palivizumab, panobacumab, P RO140, rafivirumab, raxibacumab, regavirumab, cevirumab, suvizumab, and tefibazumab; antibodies for treating hematological disorders, in particular abciximab, atorlimumab, eculizumab, mepolizumab, and milatuzumab; antibodies for immunomodulation, in particular antithymocyte globulin, basilixinab, cedelizumab, daclizumab, gavilimomab, inolinomab, muromonab-CD3, muromonab-CD3, ozumab; rimomab and siplizumab; antibodies for the treatment of diabetes, particularly gevokizumab, otelixizumab and teplizumab; antibodies for the treatment of Alzheimer's disease, particularly bapineuzumab, crenezumab, gantenerumab, ponezumab, R1450 and solanezumab; antibodies for the treatment of asthma, particularly benralizumab, enokizumab, keliximab, lebrikizumab, omalizumab, oxelumab, pascolizumab and tralokinumab;Antibodies for the treatment of various disorders, in particular brosozumab, CaroRx, fresolimumab, fulranumab, romosozumab, stamulusab, tanezumab, and ranibizumab; erythropoietin (EPO), granulocyte colony-stimulating factor (G-CSF), α-galactosidase A, α-L-iduronidase, thyrotropin α, N-acetylgalactosamine-4-sulfatase (rhASB), dornase alfa, tissue plasminogen activator (TPA) activity; These include cytopenia, glucocerebrosidase, interferon (IF) beta-1α, interferon beta-1b, interferon gamma, interferon α, TNF-α, IL-1 to IL-36, human growth hormone (rHGH), human insulin (BHI), human chorionic gonadotropin α, darbepoetin α, follicle-stimulating hormone (FSHa), and factor VIII, as well as antibodies and antibody derivatives (bispecific antibodies, multispecific antibodies, ADCs, etc.) for the treatment of various diseases. Peptides are compounds formed by the linkage of α-amino acids via peptide bonds and are intermediate products of protein hydrolysis. Peptides generally contain two to nine amino acids, and are variously called dipeptides, tripeptides, tetrapeptides, and pentapeptides depending on the number of amino acids in the peptide. Peptides consisting of three or more amino acid molecules are called polypeptides, have a molecular weight of less than 10,000 Da, and can pass through a semipermeable membrane without precipitation by trichloroacetic acid and ammonium sulfate. In some literature, peptides consisting of 2 to 10 amino acids are called oligopeptides (small peptides), peptides consisting of 10 to 50 amino acids are called polypeptides, and peptides consisting of 50 or more amino acids are called proteins. In other words, proteins are sometimes called polypeptides.

[0039] "Small molecule compounds" include 7-methoxypteridine, 7-methylpteridine, abacavir, abafungin, abarelix, acebutolol, acenaphthene, acetaminophen, acetanilide, acetazolamide, acetohexamide, etretinate, acrivastine, adenine, adenosine, alatrafloxacin, albendazole, albuterol, alclofenac, aldesleukin, alemtuzumab, alfuzosin, alitretinoin, allobarbital, allopurinol, all-trans retinoic acid (ATRA), aloxiprin, and alfuzosin. Prazolam, alprenolol, altretamine, amifostine, amiloride, aminoglutethimide, aminophenazone, amiodarone hydrochloride, amitriptyline, amlodipine, amobarbital, amodiaquine, amoxapine, amphetamine, amphotericin, amphotericin B, ampicillin, amprenavir, amsacrine, amyl nitrate, amobarbital, anastrozole, amrinone, anthracene, anthracycline antibiotics, aprobarbital, arsenic trioxide, asparaginase, aspirin, astemizole, atenolol, Atorvastatin, atovaquone, atrazine, atropine, azathioprine, auranofin, azacitidine, azapropazone, azathioprine, azintamide, azithromycin, aztreonam, baclofen, barbital, live BCG vaccine, beclamide, beclomethasone, bendroflumethiazide, benezepril, benidipine, benorylate, benperidol, bentazepam, benzamide, benzanthracene, benzathine penicillin, benzhexyl hydrochloride, benznidazole, benzodiazepine, benzoic acid, hydroxynaphtho Bephenium acetate, betamethasone, bevacizumab (atorvastatin), bexarotene, bezafibrate, bicalutamide, bifonazole, biperiden, bisacodyl, bisantrene, bleomycin, bortezomib, brinzolamide, bromazepam, bromocriptine mesylate, bromperidol, brotizolam, budesonide, bumetanide, bupropion, busulfan, butalbital, butamben, butenafine hydrochloride, butobarbital, butobarbital (butetal), butoconazole, butoconazole nitrate, butyl p-hydroxybenzoate,Caffeine, calcidiol, calciprotriene, calcitriol, calsterone, campbendazole, camphor, camptothecin, camptothecin analogues, candesartan, capecitabine, capsaicin, captopril, carbamazepine, carbimazole, carbofuran, carboplatin, carbromal, kalimazole, carmustine, cefamandole, cefazolin, cefixime, ceftazidime, cefuroxime axetil, celecoxib, cephradine, cerivastatin, cetirizine, cetuximab, chlorambucil, chloramphenicol chlordiazepoxide, chlormethiazole, chloroquine, chlorothiazide, chlorpheniramine, chlorproguanil hydrochloride, chlorpromazine, chlorpropamide, chlorprothixene, chlorpyrifos, chlortetracycline, chlorthalidone, chlorzoxazone, cholecalciferol, cilostazol, cimetidine, cinnarizine, cinoxacin, ciprofibrate, ciprofloxacin hydrochloride, cisapride, cisplatin, citalopram, cladribine, clarithromycin, clemastine fumarate, clioquinol, clobazam, Lofarabine, clofazimine, clofibrate, clomiphene citrate, clomipramine, clonazepam, clopidogrel, clotiazepam, clotrimazole, cloxacillin, clozapine, cocaine, codeine, colchicine, colistin, conjugated estrogens, corticosterone, cortisone, cortisone acetate, cyclizine, cyclobarbital, cyclobenzaprine, cyclobutane spirobarbituric acid, cycloethane-spirobarbituric acid, cycloheptane-spirobarbituric acid, cyclohexane-spirobarbituric acid, cyclopentaerythroyltransferase cyclopropane-spirobarbituric acid, cyclophosphamide, cyclopropane-spirobarbituric acid, cycloserine, cyclosporine, cyproheptadine, cyproheptadine hydrochloride, cytarabine, cytosine, dacarbazine, dactinomycin, danazol, danthron, dantrolene sodium, dapsone, darbepoetin alfa, dalodipine, daunorubicin, decoquinate, dehydroepiandrosterone, delavirdine, demeclocycline, denileukin, deoxycorticosterone, desoximetasone, dexamethasone, dextroamphetamine,Dextrochlorpheniramine, dexfenfluramine, dexrazoxane, dextropropoxyphene, heroin, amidotrizoic acid, diazepam, diazoxide, dichlorophen, dichlorprop, diclofenac, dicoumarin, didanosine, diflunisal, digitoxin, digoxin, dihydrocodeine, dihydroequiliin, dihydroergotamine mesylate, diiodohydroxyquinoline, diltiazem hydrochloride, diloxanide furoate, dimenhydrinate, dimorpholamine, dinitrumid, diosgenin, diphenoxylate hydrochloride, biphen Nil, dipyridamole, dirithromycin, disopyramide, disulfiram, diuron, docetaxel, domperidone, donepezil, doxazosin, doxazosin hydrochloride, doxorubicin (neutral), doxorubicin hydrochloride, doxycycline, dromostanolone propionate, droperidol, dyphylline, echinocandin, econazole, econazole nitrate, efavirenz, ellipticine, enalapril, enlimomab, enoximone, epinephrine, epipodophyllotoxin derivatives, epirubicin, epoetin alfa, eposartan, equinoxazone Renin, equilin, ergocalciferol, ergotamine tartrate, erlotinib, erythromycin, estradiol, estramustine, estriol, estrone, ethacrynic acid, ethambutol, etinamate, ethionamide, ethopropazine hydrochloride, ethyl 4-aminobenzoate (benzocaine), ethyl p-hydroxybenzoate, ethinylestradiol, etodolac, etomidate, etoposide, etretinate, exemestane, felbamate, felodipine, fenbendazole, fenbuconazole, fenbufen, Fenchlorphos, fenclofenac, fenfluramine, fenofibrate, fenoldepam, fenoprofen calcium, fenoxycarb, fenpiclonil, fentanyl, fenticonazole, fexofenadine, filgrastim, finasteride, flecainide acetate, floxuridine, fludarabine, fluconazole, flucytosine, fludioxonil, fludrocortisone, fludrocortisone acetate, flufenamic acid, flunanisone, flunarizine hydrochloride, flunisolide, flunitrazepam, fluocortolone,Fluometuron, fluorene, fluorouracil, fluoxetine hydrochloride, fluoxymesterone, flupentixol decanoate, flufentixol decanoate, flurazepam, flurbiprofen, fluticasone propionate, fluvastatin, folic acid, fosinopril, fosphenytoin sodium, frovatriptan, furosemide, fulvestrant, furazolidone, gabapentin, G-BHC (lindane), gefitinib, gemcitabine, gemfibrozil, gemtuzumab, glafenine, glibenclamide, gliclazide, glimepiride, glipidi , glutethimide, glibenclamide, glyceryl nitrate (nitroglycerin), goserelin acetate, grepafloxacin, griseofulvin, guaifenesin, guanabenz acetate, guanine, halofantrine hydrochloride, haloperidol, hydrochlorothiazide, heptabarbital, heroin, hesperetin, hexachlorobenzene, hexetal, histrelin acetate, hydrocortisone, hydroflumethiazide, hydroxyurea, scopolamine, hypoxanthine, ibritumomab, ibuprofen, idarubicin, arylbutylbarbituric acid, ifos Famid, ihydroequilenin, imatinib mesylate, imipenem, indapamide, indinavir, indomethacin, indoprofen, interferon alpha-2a, interferon alpha-2b, iodoamide, iopanoic acid, iprodione, irbesartan, irinotecan, isavuconazole, isocarboxazid, isoconazole, isoguanine, isoniazid, isopropylbarbituric acid, isoproturon, isosorbide dinitrate, isosorbide dinitrate, isradipine, itraconazole, itraconazole (Itra), ivermectin, ketoconazole ol, ketoprofen, ketorolac, khellin, labetalol, lamivudine, lamotrigine, lanatoside C, lanoprazole, L-DOPA, leflunomide, lenalidomide, letrozole, folic acid, leuprolide acetate, levamisole, levofloxacin, lidocaine, linuron, lisinopril, lomefloxacin, lomustine, loperamide, loratadine, lorazepam, lorefloxacin, lormetazepam, losartan mesylate, lovastatin, lisuride maleate, maprotiline hydrochloride, mazindol, mebendazole, meclozine hydrochloride, meclofenamic acid,Medazepam, methyldigoxin, medroxyprogesterone acetate, mefenamic acid, mefloquine hydrochloride, megestrol acetate, melphalan, mepenzolate bromide, meprobamate, meptazinol, purinetol, mesalazine, mesna, mesoridazine, mestranol, methadone, methaqualone, methocarbamol, mephenytoin, methotrexate, methoxsalen, methsuximide, methyclothiazide, methylphenidate, mephobarbital, methylparaben, methylprednisolone, methyltestosterone, methyprylon, methysergide maleate , metoclopramide, metolazone, metoprolol, metronidazole, mianserin hydrochloride, miconazole, midazolam, mifepristone, miglutol, minocycline, minoxidil, mitomycin C, mitotane, mitoxantrone, mycophenolate mofetil, molindone, montelukast, morphine, moxifloxacin hydrochloride, nabumetone, nadolol, nalbuphine, nalidixic acid, nandrolone, tetracene, naphthalene, naproxen, naratriptan hydrochloride, natamycin, nelarabine, nelfinavir, nevirapine, nicardipine hydrochloride niacinamide, niacin, acenocoumarol, nifedipine, nilutamide, nimodipine, nimorazole, nisodipine, nitrazepam, furantoin, furacilin, nizatidine, nofetumomab, norethindrone, norfloxacin, norgestrel, nortriptyline hydrochloride, nystatin, estradiol, ofloxacin, olanzapine, omeprazole, omoconazole, ondansetron hydrochloride, oprelvekin, ornidazole, oxaliplatin, oxamniquine, oxantel pamoate, oxaprozin, oxatomide, oxazepam Pam, oxcarbazepine, oxfendazole, oxiconazole, oxprenolol, oxyphenbutazone, oxyphencyclimine hydrochloride, paclitaxel, palifermin, pamidronic acid, p-aminosalicylic acid, pantoprazole, paramethadione, paroxetine hydrochloride, pegademase, pegaspargase, pegfilgrastim, pemetrexed disodium, penicillamine, pentaerythritol tetranitrate, pentazocine, pentobarbital, pentobarbitone, pentostatin, pentoxifylline, perphenazine,Perphenazine, pimozide, perylene, phenacemide, phenacetin, phenanthrene, phenindione, phenobarbitone, phenobarbital, phenolphthalein, phenoxybenzamine, phenoxybenzamine hydrochloride, phenoxymethylpenicillin, phensuximide, phenylbutazone, Enytoin, pindolol, pioglitazone, pipobroman, piroxicam, pizotifen maleate, platinum compounds, mithramycin, polyenoids, polymyxin B, porfimer sodium, posaconazole (Posa), pramipexole, prasterone, pravastatin, praziquantel, prazosin, prazosin hydrochloride, prednisolone, prednisone, primidone, probarbital, probenecid, probucol, procarbazine, prochlorperazine, progesterone, guanitol hydrochloride, promethazine, propofol, propoxur, Propranolol, propylparaben, propylthiouracil, prostaglandins, pseudoephedrine, pteridine-2-methylthiol, pteridine-2-thiol, pteridine-4-methylthiol, pteridine-4-thiol, pteridine-7-methylthiol, pteridine-7-thiol, pyrantel pamoate, pyrazinamide, pyrene, pyridostigmine, pyrimethamine, quetiapine, mepacrine, quinapril, quinidine, quinidine sulfate, quinine, quinine sulfate, rabeprazole sodium, ranitidine hydrochloride, rasburicase, ravuconazo steroid, repaglinide, reposal, reserpine, retinoids, rifabutin, rifampicin, rifapentine, rimexolone, risperidone, ritonavir, rituximab, rizatriptan benzoate, rofecoxib, ropinirole hydrochloride, rosiglitazone, saccharin, salbutamol, salicylamide, salicylic acid, saquinavir, sargramostim, butabarbital, secobarbital, sertaconazole, sertindole, sertraline hydrochloride, simvastatin, sirolimus, sorafenib, sparfloxacin, spiramycin, spironolactone, diazepam, Hydrotestosterone, stanozolol, stavudine, diethylstilbestrol, streptozocin, strychnine, sulconazole, sulconazole nitrate, sulfacetamide, sulfadiazine, sulfamerazine, sulfamethazine, sulfamethoxazole, sulfanilamide, sulfathiazole, sulindac, sulfabenzamide, sulfacetamide, sulfadiazine, sulfadoxine, sulfisoxazole, sulfamerazine, sulfamethoxazole, sulfapyridine, sulfasalazine, sulfinpyrazone, sulpiride,Sulthiame, sumatriptan succinate, sunitinib maleate, tacrine, tacrolimus, talbutal, tamoxifen citrate, tamulosin, targretin, taxane, tazarotene, telmisartan, temazepam, temozolomide, teniposide, tenoxicam, terazosin, terazosin hydrochloride, terbinafine hydrochloride, terbutaline sulfate, terconazole, terfenadine, testolactone, test Sterolone, tetracycline, tetrahydrocannabinol, tetroxoprim, thalidomide, thebaine, theobromine, theophylline, thiabendazole, thiamphenicol, thioguanine, thioridazine, thiotepa, totoin, thymine, tiagabine hydrochloride, tibolone, ticlopidine, tinidazole, tioconazole, tirofiban, tizanidine hydrochloride, tolazamide, tolbutamide, tolcapone, topi Lamate, topotecan, toremifene, tositumomab, tramadol, trastuzumab, trazodone hydrochloride, tretinoin, triamcinolone, triamterene, triazolam, triazole, triflupromazine, trimethoprim, trimipramine maleate, triphenylene, troglitazone, tromethamine, tropicamide, trovafloxacin, chibamate, ubidecarenone (coenzyme Q1 0), undecylenic acid, uracil, uramustine, uric acid, valproic acid, valrubicin, valsartan, vancomycin, venlafaxine hydrochloride, vigabatrin, vinbarbital, vinblastine, vincristine, vinorelbine, voriconazole, xanthine, zafirlukast, zidovudine, zileuton, zoledronic acid, zolmitriptan, zolpidem, and zopiclone.

[0040] An antigen (abbreviated Ag) is any substance capable of eliciting an immune response, triggering the production of antibodies. Foreign molecules are recognized by immunoglobulins on B cells or processed by antigen-presenting cells, where they combine with major histocompatibility complexes to generate complexes that reactivate T cells and produce a sustained immune response.

[0041] Antigenic epitopes, also known as antigenic determinants, consist of continuous sequences (primary structure of a protein) or discontinuous three-dimensional structures of proteins and determine antigenicity. Most antigenic epitopes are present on the surface of the antigenic substance, but some antigenic epitopes are present inside the antigenic substance and must be treated with enzymes or other means before they are exposed. A single natural antigenic substance can have multiple determinants, each with a wide variety of different types. The larger the antigen molecule, the greater the number of epitopes.

[0042] Tumor-associated antigens (TAA) refer to antigenic molecules present on tumor cells or normal cells, including embryonic proteins, glycoprotein antigens, squamous cell antigens, etc., and are often used in the clinical diagnosis of tumors. Tumor-associated antigens are not specific to tumor cells, but are synthesized in trace amounts on normal cells and expressed in large amounts as tumor cells proliferate, hence the term "associated antigen." Tumors derived from the same tissue type will have the same tumor-associated antigens in different individuals. Pathogenic microorganisms, also known as pathogens, are microorganisms that invade the human body and cause infections or even epidemics. Bacteria and viruses are the most harmful pathogens. Pathogenic microorganisms include prions, fungi, bacteria, spirochetes, mycoplasmas, rickettsia, chlamydia, and viruses. Pathogenic microbial antigens are substances derived from pathogens that have the function of eliciting an immune response.

[0043] "Lipid encapsulation" refers to lipid nanoparticles that provide an active substance or therapeutic agent (e.g., a nucleic acid (e.g., mRNA)) and include complete encapsulation, partial encapsulation, or both. In some embodiments, the nucleic acid (e.g., mRNA) is completely encapsulated within the lipid nanoparticle.

[0044] In various embodiments, the lipid nanoparticles have an average diameter of about 90 nm to about 600 nm, about 100 nm to about 550 nm, about 150 nm to about 500 nm, about 200 nm to about 400 nm, about 250 nm to about 300 nm, or about 200 nm to about 300 nm, and are substantially non-toxic.

[0045] B. Helper lipids In some aspects of the present disclosure, a composition comprising one or more helper lipids is mixed with the cationic lipids disclosed herein to produce lipid nanoparticles. In some embodiments, the cationic lipids are mixed with 1, 2, 3, 4, or 5 different types of helper lipids. It is contemplated that the cationic lipids can be mixed with one type of a wide variety of helper lipids. In some embodiments, the helper lipids include, but are not limited to, one or more of phospholipids, steroids or steroid derivatives, polymer-bound lipids, and modified lipids.

[0046] A "phospholipid" is a lipid containing a phosphate ester group. In some embodiments, a phospholipid is a structure comprising one or two long-chain C6-C24 alkyl or alkenyl groups, glycerol or sphingosine, one or two phosphate ester groups, and, optionally, a small organic molecule. In some embodiments, the small organic molecule is an amino acid, a sugar, or an amino-substituted alkoxy group (e.g., choline or ethanolamine). In some embodiments, the phospholipid is a phosphatidylcholine. In some embodiments, the phospholipid is DOPE, DSPC, DPPC, DMPC, DOPC, POPC, or SM. In some embodiments, the phospholipid is distearoylphosphatidylcholine or dioleoylphosphatidylethanolamine.

[0047] "Steroids and steroid derivatives" include any steroid or steroid derivative. As used herein, in some embodiments, the term "steroid" refers to a class of compounds having a tetracyclic 17-carbon ring structure that may further include one or more substituents, where the substitutions include alkyl, alkoxy, hydroxyl, oxo, and acyl, or a double bond between two or more carbon atoms. In one aspect, the steroid ring structure includes three fused cyclohexyl rings and a fused cyclopentyl ring. In some embodiments, the steroid derivative includes the above ring structure with one or more non-alkyl substituents. In some embodiments, the steroid or steroid derivative is a sterol. In some embodiments of the present disclosure, the steroid or steroid derivative is cholestane or a cholestane derivative. As described above, the cholestane derivative includes one or more non-alkyl substituents in the above ring system. In some embodiments, the cholestane or cholestane derivative is cholestane or a cholestane derivative, or a sterol or a sterol derivative. In other embodiments, the cholestane or cholestane derivative is cholestane and a sterol or a derivative thereof.

[0048] "Polymer-conjugated lipids" refer to lipids that inhibit lipid nanoparticle aggregation, improve lipid nanoparticle stability, alter immune responses, or change circulation time in vivo. Polymer-conjugated lipids include, but are not limited to, polyethylene glycol-conjugated lipids, polylactic acid-conjugated lipids, polyamide-conjugated lipids, cationic polymer-conjugated lipids, polysarcosine (pSar)-conjugated lipids, polylactic-glycolic acid (PLGA)-conjugated lipids, polyamino acid-conjugated lipids, polypeptide-conjugated lipids, and polypeptoid-conjugated lipids, or mixtures thereof. In some embodiments, "polyethylene glycol-conjugated lipids" refers to any lipid to which a PEG group is attached. In some embodiments, the PEG lipid is a diglyceride and also contains a PEG chain attached to a glycerol group. In other embodiments, the PEG lipid is a compound containing one or more C6-C24 long-chain alkyl or alkenyl groups or C6-C24 fatty acid groups attached to a linker group via a PEG chain. Non-limiting examples of PEG-lipids include PEG-modified phosphatidylethanolamines and phosphatidic acids, PEG-conjugated ceramides, PEG-modified dialkylamines, PEG-modified 1,2-diacyloxypropan-3-amines, and PEG-modified diacylglycerols and dialkylglycerols. In some embodiments, the lipid is a PEG-modified distearoylphosphatidylethanolamine or a PEG-modified dimyristoyl-sn-glycerol. In some embodiments, the PEG modification is measured by the molecular weight of the PEG component of the lipid. In some embodiments, the PEG used for modification has a molecular weight of about 100 to about 15,000. In some embodiments, the molecular weight is about 200 to about 500, about 400 to about 5000, about 500 to about 3000, or about 1200 to about 3000. The molecular weight of the PEG used for modification is from about 100, 200, 400, 500, 600, 800, 1000, 1250, 1500, 1750, 2000, 2250, 2500, 2750, 3000, 3500, 4000, 4500, 5000, 6000, 7000, 8000, 9000, 10000, or 12500 to about 15000."Modified lipids" include lipids modified with small molecule compounds, vitamins, carbohydrates, peptides, proteins, nucleic acids, lipopolysaccharides, inorganic molecules or particles, metal ions or particles, and combinations of the above.

[0049]

number

[0050] In order to more clearly explain the objects, features, and advantages of the present invention, the present invention will be described in detail below with reference to the drawings and specific embodiments. However, the embodiments of the present invention described in detail below are intended to be only illustrative of the contents of the present invention and are not intended to limit the present invention.

[0051] In the following examples, TMF1 to TMF54 refer to lipid nanoparticles formed using the compounds prepared in Example 1 as cationic lipids and other components, and are not limited to the mRNA encapsulated therein.

[0052] ALC-0315 was used as a control, and mRNA was encapsulated in a standard formulation of ALC-0315 to form lipid nanoparticles. DODAP was used as a control, and lipid nanoparticles were prepared using the same method as TMF2, except that the lipid composition was replaced with the GOLD lipids and helper lipids listed in Table 3. [Example]

[0053] Example 1: Synthesis of compounds The synthesis method of Compound 1 is as follows. [ka]

[0054] Step 1: Synthesis of Compound 1-1 DMAP (2.5 g, 20.1 mmol), DCC (4.1 g, 20.1 mmol), and DCM (50 mL) were added to a 250 mL flask and stirred until the mixture became clear. 2-Hexyldecanoic acid (5.13 g, 20.1 mmol) was added, and the mixture was stirred at room temperature for 30 minutes. Then, butyl tert-6-hydroxyhexanoate (3.6 g, 19.1 mmol) was added in one batch, and the mixture was stirred at room temperature overnight. The extent of the reaction was monitored by TLC (30% n-heptane / DCM). After completion of the reaction, the reaction solution was filtered to remove solids, and the filtrate was washed with 15 mL of water and 15 mL of saturated brine, dried over anhydrous Na2SO4, and filtered. The organic phase was concentrated to give the crude product (approximately 9.3 g). The crude product was purified by flash column chromatography (80 g silica gel, n-heptane; mobile phases were 100 mL n-heptane, 300 mL n-heptane, 300 mL 10% DCM + 90% n-heptane, and 300 mL 20% DCM + 60% n-heptane, respectively) to give the pure product (1-1, 7 g, 86% yield). 1 H NMR (400 MHz, CDCl3): δ 4.05 (m, 2H), 2.31-2.26 (m, 3H), 1.67-1.18 (m, 39H), 0.88-0.85 (m, 6H).

[0055] Step 2: Synthesis of Compound 1-2 1-1 (600 mg, 9.7 mmol) was dissolved in DCM (12 mL) and placed in a 50 mL flask. TFA (4 mL) was added. The mixture was stirred at room temperature for 2 h, and the extent of the reaction was monitored by TLC (0.5% MeOH / DCM). After completion of the reaction, the reaction solution was concentrated and dissolved in MTBE (20 mL). The pH was adjusted to 8-9 with 10% NaHCO3 solution, and then adjusted to 1-2 with 1 M dilute hydrochloric acid. The mixture was stirred for 20 min, and the layers were separated. The aqueous phase was extracted with 20 mL of MTBE, and the combined organic phases were dried over anhydrous Na2SO4, filtered, and concentrated to give the crude product (620 mg). The crude product was purified and separated by flash column (10 g silica gel packed with 50 mL of DCM; mobile phases were 100 mL of DCM, 100 mL of 1% MeOH + 99% DCM, 100 mL of 2% MeOH + 98% DCM, 100 mL of 3% MeOH + 97% DCM, and 100 mL of 4% MeOH + 96% DCM, respectively) to give the pure product (1-2, 410 mg, 78% yield). 1 H NMR (400 MHz, CDCl3): δ 4.09-4.05 (m, 2H), 2.38-2.28 (m, 3H), 1.71-1.20 (m, 30H), 0.89-0.85 (m, 6H).

[0056] Step 3: Synthesis of Compounds 1-3 1-2 (240 mg, 0.65 mmol) was dissolved in DCM (3 mL) and placed in a 25 mL flask. DMF (10 μL) was added. The reaction solution was cooled to 0-5 °C in an ice-salt bath. While maintaining the temperature of the reaction solution at 0-5 °C, oxalyl chloride (98.6 mg, 0.78 mmol) was slowly added dropwise to the solution, and the mixture was stirred at this temperature for 3 h. The extent of the reaction was monitored by TLC (20% EtOAc / n-heptane). After completion of the reaction, the reaction solution was concentrated to dryness to give the crude product (1-3, approximately 220 mg, 87% yield), which was used directly in the next step.

[0057] Step 4: Synthesis of Compound 1 3-Dimethylamino-1,2-propanediol (30.6 mg, 0.26 mmol), N,N-diisopropylethylamine (99.7 mg, 0.77 mmol), and THF (2 mL) were added to a 25 mL flask, and the reaction solution was cooled to 0-5 °C in an ice bath. A solution of 1-3 (220 mg, 0.57 mmol) in THF (2 mL) was added dropwise to the reaction solution, and the mixture was stirred at room temperature overnight. The degree of reaction was monitored by TLC (0.5% MeOH / DCM). After completion of the reaction, the reaction was quenched by adding 2 mL of water to the reaction solution, and the mixture was extracted twice with EtOAc (10 mL). The organic phases were combined, washed with 5 mL of saturated brine, dried over anhydrous Na2SO4, filtered, and concentrated to give the crude product (approximately 170 mg). The crude product was purified and separated by flash column (8 g of silica gel packed with 50 mL of n-heptane; the mobile phases were 100 mL of n-heptane, 50 mL of 1% EtOAc + 99% n-heptane; 50 mL of 2% EtOAc + 98% n-heptane; 50 mL of 3% EtOAc + 96% n-heptane; and 50 mL of 4% EtOAc + 96% n-heptane), to give the pure product (Compound 1, 40 mg, 86% yield). 1 H NMR (400 MHz, CDCl3): δ 5.22~5.18 (m, 1H), 4.38~4.34 (m, 1H), 4.10~4.04 (m, 5H), 2.49~2.43 (m, 2H), 2.35~2.27 (m, 12H), 1.69~1.25 (m, 54H), 0.89~0.85 (m, 12H).

[0058] The synthesis method of compound 2 is as follows. [ka]

[0059] Step 1: Synthesis of Compound 2 DMAP (278.4 mg, 1.35 mmol), DCC (37.5 mg, 0.31 mmol), and DCM (50 mL) were added to a 250 mL flask and stirred until the mixture became clear. 1-2 (500 mg, 1.35 mmol) was added, and the mixture was stirred at room temperature for 30 min. 3-Diethylamino-1,2-propanediol (90.3 mg, 0.61 mmol) was then added, and the mixture was stirred at room temperature for 5 h. The extent of the reaction was monitored by TLC (10% EtOAc / n-heptane). After completion of the reaction, the reaction solution was filtered to remove solids, and the filtrate was washed with 15 mL of water and 15 mL of saturated brine, respectively, and then separated. The organic phase was dried over anhydrous Na2SO4, filtered, and concentrated to give the crude product (approximately 530 mg). The crude product was purified and separated by flash column (8 g silica gel packed with 100 mL of n-heptane; mobile phases were 100 mL of 1% EtOAc + 99% n-heptane; 100 mL of 2% EtOAc + 98% n-heptane; 100 mL of 3% EtOAc + 97% n-heptane; and 500 mL of 10% EtOAc + 90% n-heptane), to give the pure product (350 mg, 71% yield). 1 H NMR (400 MHz, CDCl3): δ 5.13-5.09 (m, 1H), 4.38-4.34 (m, 1H), 4.14-4.04 (m, 5H), 2.59-2.47 (m, 6H), 2.33-2.26 (m, 6H), 1.69-1.53 (m, 13H), 1.46-1.36 (m, 9H), 1.33-1.24 (m, 43H), 0.99 (t, J=8Hz, 6H), 0.87 (t, J=4Hz, 12H).

[0060] The synthesis method of compound 3 is as follows. [ka]

[0061] Step 1: Synthesis of Compound 3-1 Pentadecan-7-ol (1.9 g, 8.32 mmol), DMAP (1.02 g, 8.32 mmol), and DCM (19 mL) were added to a 100 mL flask and stirred until the mixture became clear. Adipic anhydride (1.6 g, 12.5 mmol) was added, and the mixture was stirred at room temperature for 2 hours. The extent of the reaction was monitored by TLC (30% EtOAc / n-heptane). After completion of the reaction, the reaction solution was washed with 5 mL of water and 5 mL of saturated brine, respectively, and the layers were separated. The organic phase was dried over anhydrous Na2SO4, filtered, and concentrated to give the crude product (approximately 3.1 g). The crude product was purified and separated by flash column (60 g silica gel packed with 200 mL of n-heptane; mobile phases were 300 mL of 1% EtOAc + 99% n-heptane; 300 mL of 2% EtOAc + 98% n-heptane; 300 mL of 5% EtOAc + 95% n-heptane; and 1000 mL of 10% EtOAc + 90% n-heptane), to give the pure product (1.2 g, 40% yield). 1 H NMR (400 MHz, CDCl3): δ 4.89-4.85 (m, 1H), 2.41-2.29 (m, 4H), 1.72-1.63 (m, 4H), 1.52-1.48 (m, 4H), 1.32-1.25 (m, 20H), 0.87 (t, J=4Hz, 6H).

[0062] Step 2: Synthesis of Compound 3 DMAP (39 mg, 0.32 mmol), DCC (289 mg, 1.4 mmol), and DCM (5 mL) were added to a 250 mL flask and stirred until the mixture became clear. 3-1 (500 mg, 1.4 mmol) was added, and the mixture was stirred at room temperature for 30 min. 3-Dimethylamino-1,2-propanediol (76 mg, 0.64 mmol) was then added, and the mixture was stirred at room temperature overnight. The extent of the reaction was monitored by TLC (30% EtOAc / n-heptane). After completion of the reaction, the reaction solution was filtered to remove solids, and the filtrate was washed with 5 mL of water and 5 mL of saturated brine, respectively, and then separated. The organic phase was dried over anhydrous Na2SO4, filtered, and concentrated to give the crude product (approximately 530 mg). The crude product was purified and separated by flash column (10 g silica gel packed with 100 mL of n-heptane; mobile phases were 100 mL of 1% EtOAc+99% n-heptane; 100 mL of 2% EtOAc+98% n-heptane; 100 mL of 3% EtOAc+97% n-heptane; 100 mL of 10% EtOAc+90% n-heptane; and 500 mL of 20% EtOAc+80% n-heptane), to give the pure product (320 mg, 63% yield). 1 H NMR (400 MHz, CDCl3): δ 5.23~5.18 (m, 1H), 4.89~4.82 (m, 2H), 4.38~4.35 (m, 1H), 4.11~4.06 (m, 1H), 2.49 (s, 1H), 2.37~2.28 (m, 14H), 1.70~1.92 (m, 9H), 1.52~1.48 (m, 9H), 0.87 (t, J=4Hz, 12H).

[0063] The synthesis method of compound 4 is as follows. [ka]

[0064] Step 1: Synthesis of compound 4-1 DMAP (500.2 mg, 4.1 mmol), DCC (1.69 g, 8.2 mmol), and DCM (20 mL) were added to a 50 mL flask and stirred until the mixture became clear. 8-(tert-butoxy)-8-oxooctanoic acid (1.9 g, 8.2 mmol) was added, and the mixture was stirred at room temperature for 30 min. 9-Heptadecanol (2.1 g, 8.2 mmol) was then added in one batch, and the mixture was stirred at room temperature overnight. The extent of the reaction was monitored by TLC (10% EtOAc / n-heptane). After completion of the reaction, the reaction solution was filtered to remove solids, and the filtrate was washed with 5 mL of water and 5 mL of saturated brine, followed by separation. The organic phase was dried over anhydrous Na2SO4, filtered, and concentrated to give the crude product (approximately 3.2 g). The crude product was purified and separated by flash column (30 g of silica gel packed with 200 mL of n-heptane; the mobile phases were 200 mL of 0.5% EtOAc + 99.5% n-heptane; 200 mL of 1% EtOAc + 99% n-heptane; and 1000 mL of 2% EtOAc + 98% n-heptane), to give the pure product (2.1 g, 55% yield). 1 H NMR (400 MHz, CDCl3): δ 4.89-4.82 (m, 1H), 2.27 (t, J=8Hz, 2H), 2.19(t, J=8Hz, 2H), 1.64-1.47(m, 8H), 1.43(s, 9H), 1.36-1.25(m, 28H), 0.87(t, J=4Hz, 6H).

[0065] Step 2: Synthesis of compound 4-2 4-1 (1.2 g, 2.56 mmol) was dissolved in DCM (8 mL) and placed in a 100 mL flask. TFA (4 mL) was added. The mixture was stirred at room temperature for 2 h, and the extent of the reaction was monitored by TLC (5% MeOH / DCM). After completion of the reaction, the reaction solution was concentrated and dissolved in MTBE (20 mL). The pH was adjusted to 8-9 with 10% NaHCO3 solution, and then adjusted to 1-2 with 1 M dilute hydrochloric acid. The mixture was stirred for 20 min, and the layers were separated. The aqueous phase was extracted with 50 mL of MTBE, and the combined organic phases were dried over anhydrous Na2SO4, filtered, and concentrated to give the crude product (1.5 g). The crude product was purified and separated by flash column (100 g of silica gel packed with 50 mL of n-heptane; the mobile phases were 300 mL of 1% EtOAc + 99% n-heptane; 200 mL of 1.5% EtOAc + 98.5% n-heptane; and 1000 mL of 2% EtOAc + 98% n-heptane), to give the pure product (800 mg, 80% yield). 1 H NMR (400 MHz, CDCl3): δ 4.89-4.83 (m, 1H), 2.34 (t, J=8Hz, 2H), 2.28(t, J=8Hz, 2H), 1.67-1.59(m, 4H), 1.52-1.47(m, 4H), 1.40-1.32(m, 4H), 1.31-1.19(m, 24H), 0.87(t, J=8Hz, 6H).

[0066] Step 3: Synthesis of Compound 4 DMAP (250 mg, 1.21 mmol), DCC (33.6 mg, 0.28 mmol), and DCM (5 mL) were added to a 25 mL flask and stirred until the mixture became clear. 4-2 (500 mg, 1.21 mmol) was added, and the mixture was stirred at room temperature for 30 min. 3-Dimethylamino-1,2-propanediol (65.6 mg, 0.55 mmol) was then added in one batch, and the mixture was stirred at room temperature for 5 h. The degree of reaction was monitored by TLC (10% MeOH / DCM). After completion of the reaction, the reaction solution was filtered to remove solids, and the filtrate was washed with 10 mL of water and 10 mL of saturated brine, followed by separation. The organic phase was dried over anhydrous Na2SO4, filtered, and concentrated to give the crude product (approximately 540 mg). The crude product was purified and separated by flash column (10 g silica gel packed with 100 mL of n-heptane; mobile phases were 100 mL of 1% EtOAc + 99% n-heptane; 100 mL of 2% EtOAc + 98% n-heptane; 100 mL of 3% EtOAc + 97% n-heptane; 100 mL of 5% EtOAc + 95% n-heptane; 100 mL of 10% EtOAc + 90% n-heptane; and 300 mL of 20% EtOAc + 80% n-heptane) to give the pure product (200 mg, 40% yield). 1 H NMR (400 MHz, CDCl3): δ 5.21-5.19 (m, 1H), 4.88-4.82 (m, 2H), 4.37-4.33 (m, 1H), 4.10-4.06 (m, 1H), 2.54-2.43 (m, 2H), 2.34-2.25 (m, 14H), 1.65-1.58 (m, 9H), 1.52-1.44 (m, 9H), 1.29-1.25 (m, 52H), 0.87 (t, J=8Hz, 12H).

[0067] The synthesis method of compound 5 is as follows. [ka]

[0068] Step 1: Synthesis of compound 5-1 DMAP (1.6 g, 13.3 mmol), DCC (5.5 g, 26.6 mmol), and DCM (50 mL) were added to a 100 mL flask and stirred until the mixture became clear. 2-Hexyldecanoic acid (6.8 g, 26.6 mmol) was added, and the mixture was stirred at room temperature for 30 minutes. Then, butyl tert-6-hydroxyhexanoate (5.0 g, 26.6 mmol) was added in one batch, and the mixture was stirred at room temperature overnight. The extent of the reaction was monitored by TLC (30% n-heptane / DCM). After completion of the reaction, the reaction solution was filtered to remove solids, and the filtrate was washed with 15 mL of water and 15 mL of saturated brine. The organic phase was dried over anhydrous Na2SO4, filtered, and concentrated to give the crude product (approximately 10.2 g). The crude product was purified and separated by flash column (120 g silica gel packed with 200 mL of n-heptane; the mobile phases were 200 mL of 0.5% EtOAc + 99.5% n-heptane; and 1500 mL of 1% EtOAc + 99% n-heptane, respectively) to give the pure product (8.5 g, 75% yield).

[0069] Step 2: Synthesis of compound 5-2 1 mL of tetrahydrofuran was added to a 50 mL flask and cooled to -60 °C with liquid nitrogen. LDA (1.4 mL, 2.6 mmol) was slowly added dropwise, and the mixture was stirred for 20 minutes. While controlling the temperature below -60 °C, a mixture of 5-1 (800 mg, 1.7 mmol) and tetrahydrofuran (6 mL) was added dropwise, and the mixture was stirred for 2 hours. While controlling the temperature below -60 °C, a mixture of iodomethane (2.4 g, 17.1 mmol) and tetrahydrofuran (2 mL) was added dropwise, and the mixture was stirred for 2 hours. The degree of reaction was monitored by TLC (10% EtOAc / n-heptane). After completion of the reaction, 5 mL of saturated aqueous ammonium chloride solution was added to the reaction solution to quench the reaction, and the layers were separated. The mixture was extracted with EtOAc (20 mL × 2), and the combined organic phases were then washed with 5 mL of saturated brine, dried over anhydrous Na2SO4, and concentrated to dryness to give the crude product (approximately 900 mg). The crude product was purified and separated by flash column chromatography (25 g of silica gel, packed with 200 mL of n-heptane; the mobile phases were 200 mL of n-heptane, 200 mL of 1% EtOAc + 99% n-heptane, and 500 mL of 3% EtOAc + 97% n-heptane), giving the pure product (510 mg, 62% yield). 1 H NMR (400 MHz, CDCl3):δ 4.07-4.01(m, 2H), 2.33-2.26(m, 1H), 1.69-1.49(m, 6H), 1.44(s, 9H), 1.42-1.34(m, 5H), 1.29-1.25(m, 20H), 1.11-1.09(m, 6H), 0.89-0.85 (m, 6H).

[0070] Step 3: Synthesis of compound 5-3 5-2 (500 mg, 1.01 mmol) was dissolved in DCM (4 mL) and added to a 25 mL flask, followed by the addition of TFA (2 mL). The mixture was stirred at room temperature for 2 h, and the extent of the reaction was monitored by TLC (5% MeOH / DCM). After completion of the reaction, the reaction solution was concentrated and dissolved in MTBE (20 mL). The pH was adjusted to 8-9 with 10% NaHCO3 solution, and then adjusted to 1-2 with 1 M dilute hydrochloric acid. The mixture was stirred for 20 min, and the layers were separated. The aqueous phase was extracted with 50 mL of MTBE, and the combined organic phases were dried over anhydrous Na2SO4, filtered, and concentrated to give the crude product (400 mg). The crude product was purified and separated by flash column (8 g silica gel packed with 50 mL of n-heptane; mobile phases were 100 mL of 1% EtOAc + 99% n-heptane; 100 mL of 2% EtOAc + 98% n-heptane; 100 mL of 5% EtOAc + 95% n-heptane; and 300 mL of 10% EtOAc + 90% n-heptane), respectively, to give the pure product (280 mg, 63% yield). 1 H NMR (400 MHz, CDCl3): δ 4.13-4.03 (m, 5H), 2.50-2.42 (m, 2H), 2.04 (s, 1H), 1.77-1.55 (m, 12H), 1.51-1.33 (m, 12H), 1.30-1.24 (m, 40H), 1.19-1.18 (m, 6H), 1.09 (s, 6H), 0.87 (t, J=8Hz, 12H)

[0071] Step 4: Synthesis of compound 5-4 5-3 (280 mg, 0.7 mmol) was dissolved in DCM (3 mL) and placed in a 25 mL flask. DMF (10 μL) was added. The reaction solution was cooled to 0-5 °C in an ice-salt bath, and oxalyl chloride (177 mg, 1.4 mmol) was slowly added dropwise to the solution. The mixture was stirred at this temperature for 2 h. The degree of reaction was monitored by TLC (10% EtOAc / n-heptane). After completion of the reaction, the reaction solution was concentrated to dryness to give the crude product (approximately 265 mg, 90% yield), which was used directly in the next step.

[0072] Step 5: Synthesis of Compound 5 3-Dimethylamino-1,2-propanediol (34.4 mg, 0.29 mmol), N,N-diisopropylethylamine (112 mg, 0.87 mmol), and THF (3 mL) were added to a 25 mL flask, and the reaction solution was cooled to 0-5 °C in an ice bath. A solution of 5-4 (265 mg, 0.64 mmol) in THF (2 mL) was added dropwise to the reaction solution, and the mixture was stirred at room temperature overnight. The degree of reaction was monitored by TLC (10% MeOH / DCM). After completion of the reaction, the reaction was quenched by adding 2 mL of water to the reaction solution, and the mixture was extracted with EtOAc (10 mL × 2). The organic phases were combined, washed with 5 mL of saturated brine, dried over anhydrous Na2SO4, filtered, and concentrated to give the crude product (approximately 130 mg). The crude product was purified and separated by flash column (8 g silica gel packed with 50 mL of n-heptane; mobile phases were 100 mL of n-heptane; 100 mL of 1% EtOAc + 99% n-heptane; 100 mL of 2% EtOAc + 98% n-heptane; 100 mL of 3% EtOAc + 97% n-heptane; 100 mL of 5% EtOAc + 95% n-heptane; 100 mL of 10% EtOAc + 90% n-heptane; and 100 mL of 20% EtOAc + 80% n-heptane) to give the pure product (5, 60 mg, 24% yield). 1 H NMR (400 MHz, CDCl3): δ 5.21 (s, 1H), 4.43-4.34 (m, 1H), 4.14-3.98 (m, 5H), 2.49-2.40 (m, 4H), 2.29 (s, 6H), 1.63-1.55 (m, 12H), 1.44-1.33 (m, 12H), 1.31-1.25 (m, 40H), 1.15-1.13 (m, 6H), 0.87 (t, J=8Hz, 12H).

[0073] The synthesis method of compound 6 is as follows. [ka]

[0074] Step 1: Synthesis of compound 6-1 10 mL of tetrahydrofuran was added to a 500 mL flask and cooled to -60 °C with liquid nitrogen. LDA (34 mL, 68 mmol) was slowly added dropwise, and the mixture was stirred for 20 minutes. While controlling the temperature below -60 °C, a mixed solution of methyl oleate (10 g, 33.8 mmol) and tetrahydrofuran (90 mL) was added dropwise, and the mixture was stirred for 2 hours. While controlling the temperature below -60 °C, a mixed solution of iodomethane (48.0 g, 338 mmol) and tetrahydrofuran (10 mL) was added dropwise, and the mixture was stirred for 2 hours. The degree of reaction was monitored by TLC (n-heptane). After completion of the reaction, 50 mL of saturated aqueous ammonium chloride solution was added to the reaction solution to quench the reaction, and the liquids were separated. The mixture was extracted twice with 200 mL of EtOAc, and the combined organic phases were washed with 30 mL of saturated brine, dried over anhydrous Na2SO4, and concentrated to dryness to give the crude product (approximately 12 g). The crude product was purified and separated by flash column chromatography (100 g of silica gel, packed with 200 mL of n-heptane; the mobile phase was 2000 mL of 0.1% EtOAc + 99.9% n-heptane) to give the pure product (compound 2-1, 8.1 g, 76.1% yield). 1 H NMR (400 MHz, CDCl3): δ 5.36-5.33 (m, 2H), 3.65 (d, J=4Hz, 3H), 2.46-2.40(m, 1H), 2.04-1.98(m, 4H), 1.68-1.17(m, 22H), 1.15(d, J=4Hz, 3H), 0.88(t, J=12Hz, 3H).

[0075] Step 2: Synthesis of compound 6-2 Lithium hydroxide monohydrate (10.8 g, 257.6 mmol) and water (80 mL) were added to a 500 mL flask, followed by the addition of a mixture of 6-1 (8 g, 25.7 mmol) and THF (240 mL). The mixture was heated to 60 °C and stirred at this temperature for 6 h, then stirred at room temperature overnight. The reaction progress was monitored by TLC (30% EtOAc / n-heptane). After completion of the reaction, the THF in the reaction solution was concentrated and the pH was adjusted to 1-2 with dilute hydrochloric acid. The solution was extracted twice with EtOAc (200 mL). The combined organic phases were washed with 30 mL of saturated brine, dried over anhydrous Na2SO4, filtered, and concentrated to give the crude product (approximately 7.2 g). The crude product was purified and separated by flash column (80 g of silica gel packed with 300 mL of n-heptane; the mobile phases were 500 mL of 1% EtOAc + 99% n-heptane; 300 mL of 1.5% EtOAc + 98.5% n-heptane; and 1000 mL of 2% EtOAc + 98% n-heptane), to give the pure product (compound 2-2, 4.7 g, 62% yield). 1 H NMR (400 MHz, CDCl3): δ 5.39-5.30 (m, 2H), 4.15-4.09 (m, 1H), 2.48-2.43 (m, 1H), 2.04-1.98 (m, 4H), 1.69-1.22 (m, 22H), 1.18-1.17 (m, 3H), 0.88(t, J=12Hz, 3H).

[0076] Step 3: Synthesis of compound 6-3 The reaction product 6-2 (500 mg, 1.69 mmol) was dissolved in DCM (3 mL) and placed in a 50 mL flask. DMF (0.7 mg, 0.01 mmol) was added. The mixture was cooled to 0 °C in an ice bath, and oxalyl chloride (256.8 mg, 2.02 mmol) was added dropwise while maintaining the temperature at 0–10 °C. The extent of the reaction was monitored by TLC (30% EtOAc / n-heptane). After completion of the reaction, the reaction solution was concentrated to dryness to give the crude product (2-3, approximately 530 mg), which was used directly in the next step.

[0077] Step 4: Synthesis of Compound 6 3-Dimethylamino-1,2-propanediol (91.2 mg, 0.76 mmol), DIPEA (296.6 mg, 2.3 mmol), and THF (2 mL) were added to a 50 mL flask, and the reaction solution was cooled to -5 °C in an ice-salt bath. A solution of 6-3 (530 mg, 1.69 mmol) in THF (3 mL) was added dropwise. After the addition, the mixture was stirred at room temperature for 2 h, and the reaction progress was monitored by TLC (30% EtOAc / n-heptane). After completion of the reaction, methanol (1 mL) was added to quench the reaction. The mixture was then diluted with 10 mL of water and separated. The aqueous phase was extracted twice with 50 mL of EtOAc. The combined organic phases were washed with 5 mL of saturated brine, dried over anhydrous Na2SO4, and concentrated to dryness to give the crude product (approximately 0.7 g). The crude product was purified and separated by flash column (10 g silica gel packed with 100 mL of n-heptane; mobile phase was 500 mL of 0.1% EtOAc + 99.9% n-heptane) to give the pure product (compound 6, 300 mg, 26% yield for two steps). 1 H NMR (400 MHz, CDCl3): δ 5.35-5.32 (m, 4H), 5.20-5.18 (m, 1H), 4.39-4.36 (m, 1H), 4.11-4.08 (m, 1H), 2.46-2.39 (m, 4H), 2.26 (s, 6H), 2.03-1.98 (m, 8H), 1.67-1.62 (m, 2H), 1.43-1.25 (m, 30H), 1.13 (d, J=8Hz, 6H), 0.88 (t, J=12Hz, 6H).

[0078] The synthesis method of compound 7 is as follows. [ka]

[0079] Step 1: Synthesis of compound 7-1 5 mL of tetrahydrofuran was added to a 50 mL flask and cooled to -60 °C with liquid nitrogen. LDA (5.1 mL, 10.1 mmol) was slowly added dropwise, and the mixture was stirred for 20 minutes. While controlling the temperature below -60 °C, a mixed solution of methyl oleate (1.5 g, 5.06 mmol) and tetrahydrofuran (10 mL) was added dropwise, and the mixture was stirred for 2 hours. While controlling the temperature below -60 °C, a mixed solution of iodobutane (9.3 g, 50.6 mmol) and tetrahydrofuran (5 mL) was added dropwise, and the mixture was stirred for 2 hours. The degree of reaction was monitored by TLC (5% EtOAc / n-heptane). After completion of the reaction, 5 mL of saturated aqueous ammonium chloride solution was added to the reaction solution to quench the reaction, and the layers were separated. The mixture was extracted with EtOAc (20 mL × 2), and the combined organic phase was washed with 5 mL of saturated brine, dried over anhydrous Na2SO4, and concentrated to dryness to give the crude product (approximately 1.2 g). The crude product was purified and separated by flash column chromatography (25 g of silica gel, packed with 200 mL of n-heptane; the mobile phase was 500 mL of n-heptane) to give the pure product (compound 7-1, 850 mg, yield 47.6%). 1 H NMR (400 MHz, CDCl3): δ 5.38~5.32(m, 2H), 3.66~3.64(m, 3H), 2.36~2.27(m, 1H), 2.03~1.98(m, 4H), 1.65~1.08(m, 38H), 0.91~0.86(m, 9H).

[0080] Step 2: Synthesis of compound 7-2 A 50 mL flask was charged with 3 mL of methanol, 1.5 mL of THF, and 1 mL of sodium hydroxide (8 M / L), and 7-1 (600 mg, 1.7 mmol) was added. The mixture was heated to 70 °C and stirred at this temperature for 16 h. The reaction progress was monitored by TLC (20% EtOAc / n-heptane). After completion of the reaction, the solvent in the reaction solution was removed by concentration. The solution was diluted with water and the pH was adjusted to 1-2 with dilute hydrochloric acid. The solution was extracted with DCM (20 mL × 2). The combined organic phase was washed with 3 mL of saturated brine, dried over anhydrous Na2SO4, filtered, and concentrated to give the crude product (approximately 500 mg). The crude product was purified and separated by flash column (20 g silica gel packed with 300 mL of n-heptane; mobile phases were 100 mL of 1% EtOAc + 99% n-heptane; 100 mL of 1.5% EtOAc + 98.5% n-heptane; 100 mL of 2% EtOAc + 98% n-heptane; and 300 mL of 5% EtOAc + 95% n-heptane), to give the pure product (340 mg, 59% yield). 1 H NMR (400 MHz, CDCl3): δ 5.37~5.33 (m, 2H), 2.37~2.30 (m, 1H), 2.03~1.99 (m, 4H), 1.65~1.58 (m, 2H), 1.52~1.42 (m, 3H), 1.37~1.22 (m, 26H), 0.91~0.86 (m, 6H).

[0081] Step 3: Synthesis of compound 7-3 7-2 (340 mg, 1.0 mmol) was dissolved in DCM (5 mL) and placed in a 25 mL flask. DMF (10 μL) was added. The reaction solution was cooled to 0-5 °C in an ice-salt bath, and oxalyl chloride (152.9 mg, 1.2 mmol) was slowly added dropwise to the solution. The mixture was stirred at this temperature for 3 h. The degree of reaction was monitored by TLC (20% EtOAc / n-heptane). After completion of the reaction, the reaction solution was concentrated to dryness to give the crude product (approximately 357 mg, 100% yield), which was used directly in the next step.

[0082] Step 4: Synthesis of Compound 7 3-Dimethylamino-1,2-propanediol (54 mg, 0.45 mmol), N,N-diisopropylethylamine (175.7 mg, 1.36 mmol), and THF (2 mL) were added to a 25 mL flask, and the reaction solution was cooled to 0-5 °C in an ice bath. A solution of 7-3 (357 mg, 1.0 mmol) in THF (2 mL) was added dropwise to the reaction solution, and the mixture was stirred at room temperature overnight. The degree of reaction was monitored by TLC (0.5% MeOH / DCM). After completion of the reaction, the reaction was quenched by adding 2 mL of water to the reaction solution, and the mixture was extracted with EtOAc (10 mL × 2). The organic phases were combined, washed with 5 mL of saturated brine, dried over anhydrous Na2SO4, filtered, and concentrated to give the crude product (approximately 180 mg). The crude product was purified and separated on a flash column (8 g of silica gel, packed with 50 mL of n-heptane; the mobile phases were 100 mL of n-heptane; 50 mL of 1% EtOAc + 99% n-heptane; and 200 mL of 2% EtOAc + 98% n-heptane), to give the pure product (7, 100 mg, 29% yield). 1 H NMR (400 MHz, CDCl3): δ 5.35-5.32 (m, 4H), 5.21-5.19 (m, 1H), 4.42-4.37 (m, 1H), 4.10-4.05 (m, 1H), 2.46 (s, 2H), 2.34-2.28 (m, 7H), 2.03-1.98 (m, 8H), 1.62-1.54 (m, 4H), 1.47-1.41 (m, 6H), 1.37-1.22 (m, 58H), 0.90-0.86 (t, J=8 Hz, 12H).

[0083] Synthesis of compound 8 [ka]

[0084] Step 1: Synthesis of Compound 8 DMAP (250 mg, 1.21 mmol), DCC (33.6 mg, 0.28 mmol), and DCM (5 mL) were added to a 25 mL flask and stirred until the mixture became clear. 4-2 (500 mg, 1.21 mmol) was added, and the mixture was stirred at room temperature for 30 min. 3-Diethylamino-1,2-propanediol (80.1 mg, 0.55 mmol) was then added in one batch and stirred at room temperature for 5 h. The extent of the reaction was monitored by TLC (10% MeOH / DCM). After completion of the reaction, the reaction solution was filtered to remove solids, and the filtrate was washed with 10 mL of water and 10 mL of saturated brine, followed by separation. The organic phase was dried over anhydrous Na2SO4, filtered, and concentrated to give the crude product (approximately 600 mg). The crude product was purified and separated by flash column (10 g silica gel packed with 100 mL of n-heptane; mobile phases were 100 mL of 1% EtOAc + 99% n-heptane; 100 mL of 2% EtOAc + 98% n-heptane; 100 mL of 3% EtOAc + 97% n-heptane; 100 mL of 5% EtOAc + 95% n-heptane; 100 mL of 10% EtOAc + 90% n-heptane; and 300 mL of 20% EtOAc + 80% n-heptane) to give the pure product (230 mg, 45% yield). 1 H NMR (400 MHz, CDCl3): δ 5.12 (m, 1H), 4.89-4.83 (m, 2H), 4.37-4.33 (m, 1H), 4.14-4.10 (m, 1H), 2.53 (m, 2H), 2.32-2.25 (m, 8H), 1.65-1.58 (m, 11H), 1.52-1.48 (m, 9H), 1.39-1.25 (m, 69H), 1.00 (m, 5H), 0.87 (t, J=8Hz, 14H).

[0085] Synthesis of compound 17 [ka]

[0086] Step 1: Synthesis of compound 17-2 DMAP (74 mg, 0.61 mmol), DCC (250 mg, 1.21 mmol), and DCM (5 mL) were added to a 25 mL flask and stirred until the mixture became clear. 17-1 (500 mg, 1.21 mmol) was added, and the mixture was stirred at room temperature for 30 min. 3-Dimethylamino-1,2-propanediol (144.4 mg, 1.21 mmol) was then added in one batch and stirred at room temperature for 16 h. The extent of the reaction was monitored by TLC (5% MeOH / DCM). After completion of the reaction, the reaction solution was filtered to remove solids, and the filtrate was washed with 5 mL of water and 5 mL of saturated brine, dried over anhydrous Na2SO4, and filtered. The organic phase was concentrated to give the crude product (approximately 580 mg). The crude product was purified and separated by flash column (50 g of silica gel packed with 100 mL of DCM, mobile phases were 100 mL of 1% MeOH + 99% DCM, 100 mL of 2% MeOH + 98% DCM, 100 mL of 4% MeOH + 96% DCM, 300 mL of 10% MeOH + 90% DCM) to give the pure product (300 mg, 48% yield). 1 H NMR (400 MHz, CDCl3): δ 4.88-4.82 (m, 1H), 4.17-4.14 (m, 1H), 4.03-3.98 (m, 1H), 3.95-3.90 (m, 1H), 2.48-2.42 (m, 1H), 2.37-2.25 (m, 12H), 1.67-1.58 (m, 4H), 1.52-1.47 (m, 4H), 1.37-1.25 (m, 33H), 0.87(t, J=8Hz, 6H).

[0087] Step 2: Synthesis of compound 17 DMAP (35.7 mg, 0.29 mmol), DCC (120.5 mg, 0.58 mmol), and DCM (5 mL) were added to a 25 mL flask and stirred until the mixture became clear. 17-3 (216.4 mg, 0.58 mmol) was added, and the mixture was stirred at room temperature for 30 min. 17-2 (300 mg, 0.58 mmol) was then added in one batch, and the mixture was stirred at room temperature overnight. The extent of the reaction was monitored by TLC (30% EtOAc / n-heptane). After completion of the reaction, the reaction solution was filtered to remove solids, and the filtrate was washed with 5 mL of water and 5 mL of saturated brine, dried over anhydrous Na2SO4, and filtered. The organic phase was concentrated to give the crude product (approximately 530 mg). The crude product was purified and separated by flash column (100 g of silica gel packed with 100 mL of n-heptane; the mobile phases were 100 mL of 1% EtOAc+99% n-heptane; 100 mL of 2% EtOAc+98% n-heptane; 100 mL of 5% EtOAc+95% n-heptane; 100 mL of 10% EtOAc+90% n-heptane; and 500 mL of 30% EtOAc+70% n-heptane), to give the pure product (280 mg, 55% yield). 1 H NMR (400 MHz, CDCl3): δ 5.21-5.19 (m, 1H), 4.87-4.84 (m, 1H), 4.38-4.34 (m, 1H), 4.10-4.04 (m, 3H), 2.46 (s, 2H), 2.35-2.25 (m, 13H), 1.69-1.25 (m, 73H), 0.87(t, J=8Hz, 12H).

[0088] Synthesis of compound 18 [ka]

[0089] Step 1: Synthesis of compound 18-2 DMAP (82.4 mg, 0.67 mmol), DCC (278.4 mg, 1.35 mmol), and DCM (5 mL) were added to a 25 mL flask and stirred until the mixture became clear. 18-1 (500 mg, 1.35 mmol) was added, and the mixture was stirred at room temperature for 30 min. 3-Dimethylamino-1,2-propanediol (160.8 mg, 1.35 mmol) was then added in one batch and stirred at room temperature for 16 h. The extent of the reaction was monitored by TLC (5% MeOH / DCM). After completion of the reaction, the reaction solution was filtered to remove solids, and the filtrate was washed with 5 mL of water and 5 mL of saturated brine, dried over anhydrous Na2SO4, and filtered. The organic phase was concentrated to give the crude product (approximately 540 mg). The crude product was purified and separated by flash column (50 g silica gel packed with 100 mL of DCM, mobile phases were 100 mL of 1% MeOH + 99% DCM; 100 mL of 2% MeOH + 98% DCM; 100 mL of 4% MeOH + 96% DCM; 300 mL of 10% MeOH + 90% DCM) to give the pure product (250 mg, 39% yield). 1 H NMR (400 MHz, CDCl3): δ 4.89-4.83 (m, 1H), 2.37-2.26 (m, 4H), 1.68-1.59 (m, 4H), 1.52-1.44 (m, 4H), 1.42-1.33 (m, 4H), 1.31-1.18 (m, 25 H), 0.87(t, J=8Hz, 6H).

[0090] Step 2: Synthesis of compound 18 DMAP (34.9 mg, 0.29 mmol), DCC (118.1 mg, 0.57 mmol), and DCM (5 mL) were added to a 25 mL flask and stirred until the mixture became clear. 18-3 (236.2 mg, 0.57 mmol) was added, and the mixture was stirred at room temperature for 30 min. 18-2 (270 mg, 0.57 mmol) was then added in one batch, and the mixture was stirred at room temperature overnight. The extent of the reaction was monitored by TLC (30% EtOAc / n-heptane). After completion of the reaction, the reaction solution was filtered to remove solids, and the filtrate was washed with 5 mL of water and 5 mL of saturated brine, dried over anhydrous Na2SO4, and filtered. The organic phase was concentrated to give the crude product (approximately 550 mg). The crude product was purified and separated by flash column (100 g of silica gel packed with 100 mL of n-heptane; the mobile phases were 100 mL of 1% EtOAc+99% n-heptane; 100 mL of 2% EtOAc+98% n-heptane; 100 mL of 5% EtOAc+95% n-heptane; 100 mL of 10% EtOAc+90% n-heptane; and 500 mL of 30% EtOAc+70% n-heptane), to give the pure product (200 mg, 40% yield). 1 H NMR (400 MHz, CDCl3): δ 5.22-5.18 (m, 1H), 4.88-4.82 (m, 1H), 4.38-4.34 (m, 1H), 4.10-4.04 (m, 3H), 2.53-2.42 (m, 2H), 2.35-2.25 (m, 13H), 1.69-1.55 (m, 10H), 1.50-1.44 (m, 4H), 1.42-1.36 (m, 4H), 1.31-1.25 (m, 50H), 0.87(t, J=8Hz, 12H).

[0091] Synthesis of compound 19 [ka]

[0092] Step 1: Synthesis of compound 19 DMAP (51.4 mg, 0.42 mmol), DCC (173.6 mg, 0.84 mmol), and DCM (8 mL) were added to a 25 mL flask and stirred until the mixture became clear. 19-1 (300 mg, 0.84 mmol) was added, and the mixture was stirred at room temperature for 30 min. 17-2 (432.3 mg, 0.84 mmol) was then added in one batch, and the mixture was stirred at room temperature overnight. The extent of the reaction was monitored by TLC (30% EtOAc / n-heptane). After completion of the reaction, the reaction solution was filtered to remove solids, and the filtrate was washed with 5 mL of water and 5 mL of saturated brine, dried over anhydrous Na2SO4, and filtered. The organic phase was concentrated to give the crude product (approximately 530 mg). The crude product was purified and separated by flash column (100 g of silica gel packed with 100 mL of n-heptane; the mobile phases were 100 mL of 1% EtOAc+99% n-heptane; 100 mL of 2% EtOAc+98% n-heptane; 100 mL of 5% EtOAc+95% n-heptane; 100 mL of 10% EtOAc+90% n-heptane; and 500 mL of 30% EtOAc+70% n-heptane), to give the pure product (400 mg, 56% yield). 1 H NMR (400 MHz, CDCl3): δ 5.21-5.17 (m, 1H), 4.88-4.82 (m, 1H), 4.38-4.34 (m, 1H), 4.10-4.05 (m, 1H), 2.54-2.41 (m, 2H), 2.36-2.23 (m, 14H), 1.69-1.19 (m, 71H), 0.87(t, J=8Hz, 12H).

[0093] Synthesis of compound 20 [ka]

[0094] Step 1: Synthesis of compound 20-2 DMAP (137.1 mg, 1.12 mmol), DCC (463 mg, 2.24 mmol), and DCM (8 mL) were added to a 25 mL flask and stirred until the mixture became clear. 20-1 (800 mg, 2.24 mmol) was added, and the mixture was stirred at room temperature for 30 min. 3-Dimethylamino-1,2-propanediol (267.4 mg, 2.24 mmol) was then added in one batch, and the mixture was stirred at room temperature overnight. The extent of the reaction was monitored by TLC (30% EtOAc / n-heptane). After completion of the reaction, the reaction solution was filtered to remove solids, and the filtrate was washed with 3 mL of water and 3 mL of saturated brine, dried over anhydrous Na2SO4, and filtered. The organic phase was concentrated to give the crude product (approximately 950 mg). The crude product was purified and separated on a flash column (100 g of silica gel packed with 100 mL of n-heptane; the mobile phases were 100 mL of 99% DCM + 1% MeOH; 100 mL of 98% DCM + 2% MeOH; 100 mL of 96% DCM + 4% MeOH; and 500 mL of 92% DCM + 8% MeOH, respectively) to give the pure product (400 mg, 40% yield). 1 H NMR (400 MHz, CDCl3): δ 4.88-4.82 (m, 1H), 4.18-3.75 (m, 3H), 2.42-2.22 (m, 12H), 1.68-1.63 (m, 4H), 1.51-1.47 (m, 4H), 1.31-1.24 (m, 20H), 0.86(t, J=8Hz, 6H).

[0095] Step 2: Synthesis of compound 20 DMAP (33.4 mg, 0.27 mmol), DCC (112.7 mg, 0.55 mmol), and DCM (5 mL) were added to a 25 mL flask and stirred until the mixture became clear. 20-3 (202.4 mg, 0.55 mmol) was added, and the mixture was stirred at room temperature for 30 min. 20-2 (250 mg, 0.55 mmol) was then added in one batch, and the mixture was stirred at room temperature overnight. The extent of the reaction was monitored by TLC (10% MeOH / DCM). After completion of the reaction, the reaction solution was filtered to remove solids, and the filtrate was washed with 3 mL of water and 3 mL of saturated brine, dried over anhydrous Na2SO4, and filtered. The organic phase was concentrated to give the crude product (approximately 550 mg). The crude product was purified and separated by flash column (100 g of silica gel packed with 100 mL of n-heptane; the mobile phases were 100 mL of 1% EtOAc + 99% n-heptane; 100 mL of 2% EtOAc + 98% n-heptane; 100 mL of 5% EtOAc + 95% n-heptane; 100 mL of 10% EtOAc + 90% n-heptane; 100 mL of 30% EtOAc + 70% n-heptane; and 300 mL of 50% EtOAc + 50% n-heptane) to give the pure product (230 mg, 53% yield). 1 H NMR (400 MHz, CDCl3): δ 5.21-5.16 (m, 1H), 4.88-4.82 (m, 1H), 4.38-4.34 (m, 1H), 4.10-4.04 (m, 3H), 2.50-2.39 (m, 2H), 2.36-2.26 (m, 13H), 1.69-1.24 (m, 62H), 0.87(t, J=8Hz, 12H).

[0096] Synthesis of compound 21 [ka]

[0097] Step 1: Synthesis of compound 21-2 DMAP (137.1 mg, 1.12 mmol), DCC (463 mg, 2.24 mmol), and DCM (8 mL) were added to a 25 mL flask and stirred until the mixture became clear. 21-1 (800 mg, 2.24 mmol) was added, and the mixture was stirred at room temperature for 30 min. 3-Dimethylamino-1,2-propanediol (267.4 mg, 2.24 mmol) was then added in one batch, and the mixture was stirred at room temperature overnight. The extent of the reaction was monitored by TLC (30% EtOAc / n-heptane). After completion of the reaction, the reaction solution was filtered to remove solids, and the filtrate was washed with 3 mL of water and 3 mL of saturated brine, dried over anhydrous Na2SO4, and filtered. The organic phase was concentrated to give the crude product (approximately 950 mg). The crude product was purified and separated on a flash column (100 g of silica gel packed with 100 mL of n-heptane; the mobile phases were 100 mL of 99% DCM + 1% MeOH; 100 mL of 98% DCM + 2% MeOH; 100 mL of 96% DCM + 4% MeOH; and 500 mL of 92% DCM + 8% MeOH, respectively) to give the pure product (400 mg, 40% yield). 1 H NMR (400 MHz, CDCl3): δ 4.88-4.82 (m, 1H), 4.18-3.75 (m, 3H), 2.42-2.22 (m, 12H), 1.68-1.63 (m, 4H), 1.51-1.47 (m, 4H), 1.31-1.24 (m, 20H), 0.86(t, J=8Hz, 6H).

[0098] Step 2: Synthesis of compound 21 DMAP (33.4 mg, 0.27 mmol), DCC (112.7 mg, 0.55 mmol), and DCM (5 mL) were added to a 25 mL flask and stirred until the mixture became clear. 21-3 (225.9 mg, 0.55 mmol) was added, and the mixture was stirred at room temperature for 30 min. 21-2 (250 mg, 0.55 mmol) was then added in one batch, and the mixture was stirred at room temperature overnight. The extent of the reaction was monitored by TLC (10% MeOH / DCM). After completion of the reaction, the reaction solution was filtered to remove solids, and the filtrate was washed with 3 mL of water and 3 mL of saturated brine, dried over anhydrous Na2SO4, and filtered. The organic phase was concentrated to give the crude product (approximately 550 mg). The crude product was purified and separated by flash column (100 g of silica gel packed with 100 mL of n-heptane; the mobile phases were 100 mL of 1% EtOAc + 99% n-heptane; 100 mL of 2% EtOAc + 98% n-heptane; 100 mL of 5% EtOAc + 95% n-heptane; 100 mL of 10% EtOAc + 90% n-heptane; 100 mL of 30% EtOAc + 70% n-heptane; and 400 mL of 50% EtOAc + 50% n-heptane) to give the pure product (340 mg, 73% yield). 1 H NMR (400 MHz, CDCl3): δ 5.21-5.16 (m, 1H), 4.88-4.82 (m, 1H), 4.38-4.33 (m, 1H), 4.10-4.05 (m, 1H), 2.50-2.40 (m, 2H), 2.36-2.21 (m, 14H), 1.68-1.58 (m, 8H), 1.52-1.47 (m, 8H), 1.38-1.25 (m, 52H), 0.87 (t, J=8Hz, 12H).

[0099] Synthesis of compound 22 [ka]

[0100] Step 1: Synthesis of compound 22 DMAP (51.4 mg, 0.42 mmol), DCC (173.6 mg, 0.84 mmol), and DCM (8 mL) were added to a 25 mL flask and stirred until the mixture became clear. 22-2 (300 mg, 0.84 mmol) was added, and the mixture was stirred at room temperature for 30 min. 22-1 (396.9 mg, 0.84 mmol) was then added in one batch, and the mixture was stirred at room temperature overnight. The extent of the reaction was monitored by TLC (30% EtOAc / n-heptane). After completion of the reaction, the reaction solution was filtered to remove solids, and the filtrate was washed with 5 mL of water and 5 mL of saturated brine, dried over anhydrous Na2SO4, and filtered. The organic phase was concentrated to give the crude product (approximately 530 mg). The crude product was purified and separated by flash column (100 g of silica gel packed with 100 mL of n-heptane; the mobile phases were 100 mL of 1% EtOAc+99% n-heptane; 100 mL of 2% EtOAc+98% n-heptane; 100 mL of 5% EtOAc+95% n-heptane; 100 mL of 10% EtOAc+90% n-heptane; and 500 mL of 30% EtOAc+70% n-heptane), to give the pure product (430 mg, 63% yield). 1 H NMR (400 MHz, CDCl3): δ 5.21 (s, 1H), 4.89-4.83 (m, 1H), 4.39-4.35 (m, 1H), 4.10-4.04 (m, 3H), 2.46 (s, 2H), 2.36-2.27 (m, 12H), 1.70-1.25 (m, 65H), 0.87(t, J=8Hz, 12H).

[0101] Synthesis of compound 23 [ka]

[0102] Step 1: Synthesis of compound 23-2 5 mL of tetrahydrofuran was added to a 50 mL flask and cooled to -60 °C with liquid nitrogen. LDA (16.86 mL, 33.7 mmol) was slowly added dropwise, and the mixture was stirred for 20 minutes. While maintaining the temperature below -60 °C, a mixed solution of methyl oleate (23-1, 5.0 g, 16.86 mmol) and tetrahydrofuran (20 mL) was added dropwise, and the mixture was stirred for 2 hours. While maintaining the temperature below -60 °C, a mixed solution of 2-ethyl iodoethane (8.1 g, 33.7 mmol) and tetrahydrofuran (5 mL) was added dropwise, and the mixture was slowly warmed to room temperature and stirred for 16 hours. The degree of reaction was monitored by TLC (n-heptane). After completion of the reaction, 5 mL of saturated aqueous ammonium chloride solution was added to the reaction solution to quench the reaction, and the liquids were separated. The mixture was extracted with EtOAc (20 mL × 2), and the combined organic phases were washed with 5 mL of saturated brine, dried over anhydrous Na2SO4, and concentrated to dryness to give the crude product (approximately 5.0 g). The crude product was purified and separated by flash column chromatography (50 g of silica gel packed with 200 mL of n-heptane; the mobile phases were 300 mL of n-heptane, 300 mL of 0.1% EtOAc + 99.9% n-heptane, and 500 mL of 0.2% EtOAc + 99.8% n-heptane), affording the pure product (3.0 g, 43.5% yield).

[0103] Step 2: Synthesis of compound 23-3 Methanol (15 mL), THF (12 mL), and 4.6 mL of sodium hydroxide (8 mM) were added to a 100 mL flask, and 23-2 (3.0 g, 7.34 mmol) was added. The mixture was heated to 65 °C and stirred for 16 h. The reaction progress was monitored by TLC (20% EtOAc / n-heptane). After completion of the reaction, the solvent in the reaction solution was removed by concentration. The solution was diluted with water and the pH was adjusted to 1-2 with dilute hydrochloric acid. The solution was extracted with DCM (20 mL × 2), and the combined organic phase was washed with 3 mL of saturated brine, dried over anhydrous Na2SO4, filtered, and concentrated to give the crude product (approximately 1.5 g). The crude product was purified and separated on a flash column (60 g of silica gel packed with 300 mL of n-heptane; the mobile phases were 100 mL of 1% EtOAc + 99% n-heptane; 100 mL of 1.5% EtOAc + 98.5% n-heptane; 100 mL of 2% EtOAc + 98% n-heptane; 300 mL of 5% EtOAc + 95% n-heptane; and 300 mL of 10% EtOAc + 90% n-heptane), to give the pure product (650 mg, 22% yield).

[0104] Step 3: Synthesis of compound 23-4 23-3 (100 mg, 0.25 mmol) was dissolved in DCM (1 mL) and placed in a 25 mL flask. DMF (10 μL) was added. The reaction solution was cooled to 0-5 °C in an ice-salt bath, and oxalyl chloride (51.5 mg, 0.41 mmol) was slowly added dropwise to the solution. The mixture was stirred at this temperature for 3 h. The degree of reaction was monitored by TLC (20% EtOAc / n-heptane). After completion of the reaction, the reaction solution was concentrated to dryness to give the crude product (approximately 104 mg, 100% yield), which was used directly in the next step.

[0105] Step 4: Synthesis of compound 23 3-Dimethylamino-1,2-propanediol (13.6 mg, 0.11 mmol), N,N-diisopropylethylamine (44.4 mg, 0.34 mmol), and THF (1 mL) were added to a 25 mL flask, and the reaction solution was cooled to 0-5 °C in an ice bath. A solution of 23-4 (104 mg, 0.25 mmol) in THF (2 mL) was added dropwise to the reaction solution, and the mixture was stirred at room temperature overnight. The degree of reaction was monitored by TLC (0.5% MeOH / DCM). After completion of the reaction, the reaction was quenched by adding 2 mL of water to the reaction solution, and the mixture was extracted with EtOAc (10 mL × 2). The organic phases were combined, washed with 5 mL of saturated brine, dried over anhydrous Na2SO4, filtered, and concentrated to give the crude product (approximately 180 mg). The crude product was purified and separated by flash column (8 g of silica gel, packed with 50 mL of n-heptane; mobile phases were 100 mL of n-heptane; 100 mL of 1% EtOAc + 99% n-heptane; 100 mL of 2% EtOAc + 98% n-heptane; 100 mL of 3% EtOAc + 97% n-heptane; 100 mL of 5% EtOAc + 95% n-heptane; and 200 mL of 10% EtOAc + 90% n-heptane) to give the pure product (C021, 40 mg, 40% yield). 1 H NMR (400 MHz, CDCl3): δ5.36~5.33(m, 4H), 2.46~2.39(m, 2H), 2.03~1.98(m, 8H), 1.64~1.58(m, 4H), 1.49~1.19(m, 66H), 0.91~0.81(m, 18H).

[0106] Synthesis of compound 24 [ka]

[0107] Step 1: Synthesis of compound 24-2 2 mL of tetrahydrofuran was added to a 50 mL flask and cooled to -60 °C with liquid nitrogen. LDA (7.5 mL, 14.9 mmol) was slowly added dropwise, and the mixture was stirred for 20 minutes. While maintaining the temperature below -60 °C, a mixed solution of methyl palmitoleate (24-1, 2.0 g, 7.5 mmol) and tetrahydrofuran (5 mL) was added dropwise, and the mixture was stirred for 2 hours. While maintaining the temperature below -60 °C, a mixed solution of 2-ethyl iodoethane (4.8 g, 20.1 mmol) and tetrahydrofuran (1 mL) was added dropwise, and the mixture was slowly warmed to room temperature and stirred for 16 hours. The degree of reaction was monitored by TLC (n-heptane). After completion of the reaction, 5 mL of saturated aqueous ammonium chloride solution was added to the reaction solution to quench the reaction, and the liquids were separated. The mixture was extracted with EtOAc (20 mL × 2), and the combined organic phases were washed with 5 mL of saturated brine, dried over anhydrous Na2SO4, and concentrated to dryness to give the crude product (approximately 1.6 g). The crude product was purified and separated by flash column chromatography (50 g of silica gel packed with 200 mL of n-heptane; mobile phases were 200 mL of n-heptane, 200 mL of 0.2% EtOAc + 99.8% n-heptane, 200 mL of 0.3% EtOAc + 99.7% n-heptane, and 300 mL of 0.4% EtOAc + 99.6% n-heptane) to give the pure product (600 mg, 21.2% yield).

[0108] Step 2: Synthesis of compound 24-3 Methanol (4 mL), THF (6 mL), and sodium hydroxide (0.98 mL, 8 mM) were added to a 100 mL flask, and 24-2 (600 mg, 1.58 mmol) was added. The mixture was heated to 65 °C and stirred for 16 h. The reaction progress was monitored by TLC (20% EtOAc / n-heptane). After completion of the reaction, the solvent in the reaction solution was removed by concentration. The solution was diluted with water and the pH was adjusted to 1-2 with dilute hydrochloric acid. The solution was extracted with DCM (20 mL × 2), and the combined organic phase was washed with 3 mL of saturated brine, dried over anhydrous Na2SO4, filtered, and concentrated to give the crude product (approximately 800 mg). The crude product was purified and separated by flash column (30 g silica gel packed with 100 mL of n-heptane; mobile phases were 100 mL of 1% EtOAc + 99% n-heptane; 100 mL of 1.5% EtOAc + 98.5% n-heptane; 100 mL of 2% EtOAc + 98% n-heptane; 100 mL of 5% EtOAc + 95% n-heptane; and 300 mL of 10% EtOAc + 90% n-heptane), to give the pure product (300 mg, 52% yield).

[0109] Step 3: Synthesis of compound 24-4 24-3 (300 mg, 0.82 mmol) was dissolved in DCM (3 mL) and placed in a 25 mL flask. DMF (10 μL) was added. The reaction solution was cooled to 0-5 °C in an ice-salt bath, and oxalyl chloride (166.2 mg, 1.31 mmol) was slowly added dropwise to the solution. The mixture was stirred at this temperature for 3 h. The degree of reaction was monitored by TLC (20% EtOAc / n-heptane). After completion of the reaction, the reaction solution was concentrated to dryness to give the crude product (approximately 315 mg, 100% yield), which was used directly in the next step.

[0110] Step 4: Synthesis of compound 24 3-Dimethylamino-1,2-propanediol (44.3 mg, 0.37 mmol), N,N-diisopropylethylamine (114.2 mg, 1.2 mmol), and THF (1 mL) were added to a 25 mL flask, and the reaction solution was cooled to 0-5 °C in an ice bath. A solution of 24-4 (315 mg, 0.82 mmol) in THF (2 mL) was added dropwise to the reaction solution, and the mixture was stirred at room temperature overnight. The degree of reaction was monitored by TLC (0.5% MeOH / DCM). After completion of the reaction, the reaction was quenched by adding 2 mL of water to the reaction solution, and the mixture was extracted with EtOAc (10 mL × 2). The organic phases were combined, washed with 5 mL of saturated brine, dried over anhydrous Na2SO4, filtered, and concentrated to give the crude product (approximately 180 mg). The crude product was purified and separated by flash column (8 g of silica gel, packed with 50 mL of n-heptane; mobile phases were 100 mL of n-heptane; 100 mL of 1% EtOAc + 99% n-heptane; 100 mL of 2% EtOAc + 98% n-heptane; 100 mL of 4% EtOAc + 96% n-heptane; and 300 mL of 6% EtOAc + 94% n-heptane) to give the pure product (C022, 150 mg, 50% yield). 1 H NMR (400 MHz, CDCl3): δ 5.38~5.29 (m, 4H), 5.18 (s, 1H), 4.39~4.35 (m, 1H), 4.10~4.05 (m, 1H), 2.45~2.38 (m, 4H), 2.27 (s, 5H), 2.03~1.98 (m, 8H), 1.62~1.54 (m, 4H), 1.44~1.14 (m, 56H), 0.90~0.79 (m, 18H).

[0111] Synthesis of compound 25 [ka]

[0112] Step 1: Synthesis of compound 25-3 N-Methylethanolamine (25-1, 3.4 g, 45.3 mmol) and DCM (50 mL) were added to a 500 mL flask and cooled to 0-5 °C in an ice-salt bath. To the cooled solution, a solution of TBDPSCl (25-2, 12.44 g, 45.3 mmol) in DCM (50 mL) was slowly added dropwise. After the addition, the mixture was stirred at room temperature for 2 h. The reaction progress was monitored by TLC (10% MeOH / DCM). After completion of the reaction, the reaction was quenched by adding 30 mL of water and the layers were separated. The aqueous phase was extracted with DCM (50 mL × 3). The combined organic phase was washed with 50 mL of saturated brine, dried over anhydrous Na2SO4, filtered, and concentrated to give the crude product (approximately 12 g). The crude product was purified and separated by flash column chromatography (200 g of silica gel packed with 300 mL of n-heptane; the mobile phases were 300 mL of n-heptane; 300 mL of 0.1% EtOAc + 99.9% n-heptane; 300 mL of 0.3% EtOAc + 99.7% n-heptane; 300 mL of 1% EtOAc + 99% n-heptane; 300 mL of 1% EtOAc + 99% n-heptane; 300 mL of 0.2% MeOH + 99.8% DCM; 500 mL of 0.3% MeOH + 99.7% DCM) to give the pure product (9 g, 63% yield).

[0113] Step 2: Synthesis of compound 25-5 25-3 (5.0 g, 16 mmol), potassium carbonate (5.5 g, 39.9 mmol), and 3-chloro-1,2-propanediol (25-4, 2.12 g, 19.1 mmol) were dissolved in isopropanol (15 mL) and added to a 100 mL flask. The mixture was heated to 80 °C and stirred overnight. The extent of the reaction was monitored by TLC (10% MeOH / DCM). After completion of the reaction, the mixture was filtered, and the organic phase was concentrated to give the crude product (approximately 10.1 g). The crude product was purified and separated by flash column (100 g silica gel, 200 mL of 0.1% MeOH+99.9% DCM; 200 mL of 0.2% MeOH+99.8% DCM; 200 mL of 0.3% MeOH+99.7% DCM; 200 mL of 0.5% MeOH+99.5% DCM; and 300 mL of 1% MeOH+99% DCM) to give the pure product (3.6 g, 58% yield).

[0114] Step 3: Synthesis of compound 25-7 25-6 (1.0 g, 2.7 mmol), DCC (556.8 mg, 2.7 mmol), DMAP (82.4 mg, 0.67 mmol), and DCM (3 mL) were added to a 50 mL flask. After stirring the mixture at room temperature for 30 min, 25-5 (523 mg, 1.35 mmol) was added to the reaction solution. The reaction extent was monitored by TLC (30% EtOAc / n-heptane). After completion of the reaction, the reaction solution was filtered to remove solids, and the filtrate was washed with 5 mL of water and 5 mL of saturated brine, dried over anhydrous Na2SO4, and filtered. The organic phase was concentrated to give the crude product (approximately 1.5 g). The crude product was purified and separated by flash column (50 g silica gel; mobile phases were 200 mL n-heptane; 150 mL 1% EtOAc + 99% n-heptane; 150 mL 2% EtOAc + 98% n-heptane; 200 mL 5% EtOAc + 95% n-heptane; and 500 mL 8% EtOAc + 92% n-heptane) to give the pure product (1.2 g, 81% yield).

[0115] Step 4: Synthesis of compound 25 25-7 (1.0 g, 0.92 mmol) was dissolved in THF (5 mL) and added to a 50 mL flask. The mixture was cooled to -5 °C in an ice-salt bath. TBAF (717.9 mg, 2.75 mmol) was then added, and the mixture was stirred at room temperature overnight. The reaction progress was monitored by TLC (10% MeOH / DCM). After completion of the reaction, 5 mL of aqueous ammonium chloride solution was added to quench the reaction, and the layers were separated. The aqueous phase was extracted with EtOAc (10 mL × 3). The combined organic phase was washed with 5 mL of saturated brine, dried over anhydrous Na2SO4, filtered, and concentrated to give the crude product (approximately 700 mg). The crude product was purified and separated by flash column (12 g silica gel; mobile phases were 300 mL of n-heptane; 200 mL of 1% EtOAc+99% n-heptane; 200 mL of 5% EtOAc+95% n-heptane; 200 mL of 8% EtOAc+92% n-heptane; 200 mL of 15% EtOAc+85% n-heptane; and 400 mL of 30% EtOAc+70% n-heptane) to give the pure product (350 mg, 45% yield). 1H NMR (400 MHz, CDCl3): δ 5.23-5.19 (m, 1H), 4.36-4.32 (m, 1H), 4.12-4.04 (m, 5H), 3.58 (t, J=4Hz, 2H), 2.66-2.56 (m, 4H), 2.36-2.26 (m, 9H), 1.69~1.53 (m, 13H), 1.46~1.35 (m, 8H), 1.32~1.19 (m, 43H), 0.88~0.85 (m, 12H).

[0116] Synthesis of compound 27 [ka]

[0117] Step 1: Synthesis of compound 27-2 Benzaldehyde (5.91 g, 55.66 mmol) and ethanol (17 mL) were added to a 100 mL flask and cooled to 18 °C. Ammonium hydroxide (11.97 g, 85.39 mmol) was slowly added dropwise to the cooled solution. After the addition, the mixture was stirred at 18 °C for 2 hours. Next, epichlorohydrin (5 g, 54.04 mmol) was weighed and added, and the mixture was stirred at 40-45 °C for 16 hours. After the reaction was completed, the reaction solution was concentrated to dryness, and toluene (20 mL) and water (20 mL) were added to dissolve the crude product. Concentrated hydrochloric acid (8.32 g, 82.13 mmol) was weighed and added dropwise to the reaction system. The mixture was stirred for 3 hours while maintaining the temperature at 35-40 °C, and then the layers were separated. The toluene phase was washed with water (10 mL). The aqueous phases were combined and concentrated to obtain the product (6 g).

[0118] Step 2: Synthesis of compound 27-3 27-2 (3 g, 20.55 mmol), di-tert-butyl dicarbonate (4.89 g, 22.40 mmol), methanol (12 mL), and water (7 mL) were added to a 100 mL flask. Potassium bicarbonate (2.26 g, 11.60 mmol) was then weighed and added to the reaction solution in three batches, and the mixture was stirred for 3 hours. The degree of reaction was monitored by TLC (10% MeOH / DCM). After completion of the reaction, dichloromethane (15 mL) was added, and the mixture was extracted with water (6 mL) and then separated. The organic phase was concentrated to obtain the crude product (approximately 5 g). The crude product was purified and separated by flash column (20 g silica gel packed with 100 mL of n-heptane; 100 mL of 1% EtOAc + 99% n-heptane; 100 mL of 5% EtOAc + 95% n-heptane; and 600 mL of 10% EtOAc + 90% n-heptane) to give the pure product (800 mg, 19% yield).

[0119] Step 3: Synthesis of compound 27-5 27-3 (700 mg, 3.34 mmol), 27-4 (1.05 g, 3.34 mmol), potassium carbonate (1.15 g, 8.35 mmol), and isopropanol (8 mL) were added to a 25 mL flask, and the mixture was heated to 80 °C and stirred for 16 h. The reaction extent was monitored by TLC (10% MeOH / DCM). After completion of the reaction, the mixture was filtered, and the organic phase was concentrated to give the crude product (approximately 2 g). The crude product was purified and separated by flash column (12 g silica gel; mobile phases were 100 mL each of n-heptane; 100 mL of 1% EtOAc+99% n-heptane; 100 mL of 2% EtOAc+98% n-heptane; 100 mL of 8% EtOAc+92% n-heptane; 100 mL of 15% EtOAc+85% n-heptane; and 600 mL of 30% EtOAc+70% n-heptane) to give the pure product (1 g, 62% yield). 1H NMR (400 MHz, CDCl3): δ 7.68~7.66 (m, 4H), 7.43~7.37 (m, 6H), 4.99 (m, 1H), 3.74~3.70 (m, 3H), 3.35~3.31 (m, 1H), 3.03~2.97 (m, 1H), 2.73~2.67 (m, 1H), 2.55~2.52 (m, 1H), 2.41~2.38 (m, 2H), 2.30 (m, 3H), 1.44 (s, 9H), 1.06~1.04 (m, 9H).

[0120] Step 4: Synthesis of compound 27-6 27-5 (300 mg, 0.62 mmol) and EtOAc (2 mL) were added to a 25 mL flask, and the mixture was cooled to 0–5 °C in an ice bath. HCl / EtOAc (0.6 mL, 4 M) was weighed and added, and the mixture was warmed to room temperature and stirred for 3 h. The extent of the reaction was monitored by TLC (10% MeOH / DCM). After completion of the reaction, the organic phase was concentrated to give the product (approximately 300 mg).

[0121] Step 5: Synthesis of compound 27-8 27-7 (496.14 mg, 1.34 mmol), DCC (276.25 mg, 1.34 mmol), DMAP (122.68 mg, 1.0 mmol), and DCM (5 mL) were added to a 25 mL flask, and the mixture was stirred at room temperature for 30 min. 27-6 (330 mg, 0.66 mmol) was then added dropwise, and the mixture was stirred at room temperature for 16 h. The extent of the reaction was monitored by TLC (30% EtOAc / n-heptane). After completion of the reaction, the mixture was filtered, and the filtrate was washed with 5 mL of water and 5 mL of saturated brine, dried over anhydrous Na2SO4, and filtered. The organic phase was concentrated to give the crude product (approximately 1 g). The crude product was purified and separated by flash column (12 g silica gel; mobile phases were 100 mL of n-heptane; 100 mL of 1% EtOAc + 99% n-heptane; 100 mL of 2% EtOAc + 98% n-heptane; 100 mL of 8% EtOAc + 92% n-heptane; and 200 mL of 20% EtOAc + 80% n-heptane) to give the pure product (330 mg, 45% yield). 1H NMR (400 MHz, CDCl3): δ 7.67~7.65 (m, 4H), 7.45~7.36 (m, 6H), 6.26 (m, 1H), 4.98 (m, 1H), 4.07~4.02 (m, 4H), 3.75 (m, 2H), 3.49~3.45 (m, 2H), 2.59 (m, 4H), 2.34~2.22 (m, 7H), 2.11~2.08 (m, 2H), 1.67~1.53 (m, 14H), 1.46~1.25 (m, 60H), 1.05 (s, 9H), 0.89~0.85 (m, 12H).

[0122] Step 6: Synthesis of compound 27 27-8 (330 mg, 0.31 mmol) and THF (3 mL) were added to a 25 mL flask and cooled to 0–5 °C in an ice bath. TBAF (240.19 mg, 0.92 mmol) was added dropwise while maintaining the temperature at 0–5 °C, and the mixture was stirred at this temperature for 3 h. The extent of the reaction was monitored by TLC (5% MeOH / DCM). After completion of the reaction, the reaction was quenched with saturated aqueous ammonium chloride solution, and the mixture was extracted with EtOAc (5 mL × 2). The organic phases were combined, dried over anhydrous Na2SO4, filtered, and concentrated to give the crude product (approximately 500 mg). The crude product was purified and separated by flash column (4 g of silica gel packed with 100 mL of n-heptane; the mobile phases were 100 mL of 1% EtOAc+99% n-heptane; 100 mL of 2% EtOAc+98% n-heptane; 100 mL of 10% EtOAc+90% n-heptane; 100 mL of 30% EtOAc+70% n-heptane; and 300 mL of 50% EtOAc+50% n-heptane), to give the pure product (100 mg, 39% yield). 1 H NMR (400 MHz, CDCl3): δ 6.14~6.11 (m, 1H), 5.06~5.04 (m, 1H), 4.08~4.04 (m, 4H), 3.63~3.46 (m, 4H), 2.64~2.60 (m, 4H), 2.35~2.26 (m, 7H), 2.20~2.17 (m, 2H), 1.70~1.53 (m, 12H), 1.46~1.35 (m, 8H), 1.33~1.25 (m, 45H), 0.89~0.85 (m, 12H).

[0123] Synthesis of compound 29 [ka]

[0124] Step 1: Synthesis of compound 29-2 Pentadecan-7-ol (29-1, 1.0 g, 4.4 mmol), succinic anhydride (525.7 mg, 5.3 mmol), DMAP (267.4 mg, 2.2 mmol), and DCM (5 mL) were added to a 50 mL flask, and the mixture was stirred at room temperature overnight. The extent of the reaction was monitored by TLC (30% EtOAc / n-heptane). After completion of the reaction, the mixture was washed with 5 mL of water and 5 mL of saturated brine, dried over anhydrous Na2SO4, and filtered. The organic phase was concentrated to give the crude product (approximately 800 mg). The crude product was purified and separated by flash column (50 g silica gel; mobile phases were 200 mL n-heptane; 150 mL 1% EtOAc+99% n-heptane; 150 mL 2% EtOAc+98% n-heptane; 200 mL 8% EtOAc+92% n-heptane; 200 mL 20% EtOAc+80% n-heptane; and 500 mL 35% EtOAc+70% n-heptane) to give the pure product (540 mg, 38% yield).

[0125] Step 2: Synthesis of compound 29 DMAP (46.5 mg, 0.38 mmol), DCC (314.1 mg, 1.52 mmol), and DCM (5 mL) were added to a 25 mL flask and stirred until the mixture became clear. 29-2 (500 mg, 1.52 mmol) was added, and the mixture was stirred at room temperature for 30 min. 3-Dimethylamino-1,2-propanediol (90.7 mg, 0.76 mmol) was then added in one batch, and the mixture was stirred at room temperature for 5 h. The extent of the reaction was monitored by TLC (10% MeOH / DCM). After completion of the reaction, the reaction solution was filtered to remove solids, and the filtrate was washed with 10 mL of water and 10 mL of saturated brine, dried over anhydrous Na2SO4, and filtered. The organic phase was concentrated to give the crude product (approximately 510 mg). The crude product was purified and separated by flash column (20 g silica gel, packed with 100 mL of n-heptane; mobile phases were 100 mL of 1% EtOAc+99% n-heptane; 100 mL of 2% EtOAc+98% n-heptane; 100 mL of 3% EtOAc+97% n-heptane; 100 mL of 5% EtOAc+95% n-heptane; 100 mL of 10% EtOAc+90% n-heptane; 100 mL of 20% EtOAc+80% n-heptane; 300 mL of 30% EtOAc+70% n-heptane) to give the pure product (260 mg, 46% yield).

[0126] Synthesis of compound 30 [ka]

[0127] Step 1: Synthesis of compound 30-2 2-Hexyldecanoic acid (30-1, 1.0 g, 3.9 mmol), DMAP (238.2 mg, 1.95 mmol), DCC (804.6 mg, 3.9 mmol), and DCM (5 mL) were added to a 50 mL flask, and the mixture was stirred at room temperature for 30 min. Butyl tert-4-hydroxybutyrate (624.8 mg, 3.9 mmol) was added, and the mixture was stirred at room temperature overnight. The extent of the reaction was monitored by TLC (20% EtOAc / n-heptane). After completion of the reaction, the mixture was filtered, and the filtrate was washed with 5 mL of water and 5 mL of saturated brine, dried over anhydrous Na2SO4, and filtered. The organic phase was concentrated to give the crude product (approximately 2.0 g). The crude product was purified and separated by flash column (50 g silica gel; mobile phases were 200 mL n-heptane; 150 mL 1% EtOAc + 99% n-heptane; 150 mL 1.5% EtOAc + 98.5% n-heptane; 150 mL 2% EtOAc + 98% n-heptane; and 500 mL 3% EtOAc + 97% n-heptane) to give the pure product (1.5 g, 96% yield).

[0128] Step 2: Synthesis of compound 30-3 30-2 (1.5 g, 3.76 mmol) was dissolved in DCM (6 mL) and placed in a 100 mL flask. TFA (3 mL) was added. The mixture was stirred at room temperature for 2 h, and the extent of the reaction was monitored by TLC (5% MeOH / DCM). After completion of the reaction, the reaction solution was concentrated and dissolved in MTBE (20 mL). The pH was adjusted to 8-9 with 10% NaHCO3 solution, and then adjusted to 1-2 with 1 M dilute hydrochloric acid. The mixture was stirred for 20 min, and the layers were separated. The aqueous phase was extracted with MTBE (50 mL). The combined organic phases were dried over anhydrous Na2SO4, filtered, and concentrated to give the crude product (1.5 g). The crude product was purified and separated by flash column (100 g of silica gel packed with 50 mL of n-heptane; the mobile phases were 200 mL of 1% EtOAc + 99% n-heptane; 200 mL of 2% EtOAc + 98% n-heptane; 200 mL of 5% EtOAc + 95% n-heptane; and 500 mL of 10% EtOAc + 90% n-heptane), respectively, to give the pure product (1 g, 77% yield). 1H NMR (400 MHz, CDCl3): δ 4.12 (t, J=4Hz, 2H), 2.46 (t, J=4Hz, 2H), 2.35~2.28 (m, 1H), 2.01~1.94 (m, 2H), 1.62~1.53 (m, 2H), 1.47~1.39 (m, 2H), 1.30~1.20 (m, 20H), 0.88~0.85 (m, 6H).

[0129] Step 3: Synthesis of Compound 30 DMAP (44.6 mg, 0.36 mmol), DCC (301.2 mg, 1.46 mmol), and DCM (5 mL) were added to a 25 mL flask and stirred until the mixture became clear. 30-3 (500 mg, 1.46 mmol) was added, and the mixture was stirred at room temperature for 30 min. 3-Dimethylamino-1,2-propanediol (86.9 mg, 0.73 mmol) was then added in one batch, and the mixture was stirred at room temperature for 5 h. The extent of the reaction was monitored by TLC (30% EtOAc / n-heptane). After completion of the reaction, the reaction solution was filtered to remove solids, and the filtrate was washed with 5 mL of water and 5 mL of saturated brine, dried over anhydrous Na2SO4, and filtered. The organic phase was concentrated to give the crude product (approximately 510 mg). The crude product was purified and separated by flash column (20 g silica gel packed with 100 mL of n-heptane; mobile phases were 100 mL of 1% EtOAc + 99% n-heptane; 100 mL of 2% EtOAc + 98% n-heptane; 100 mL of 3% EtOAc + 97% n-heptane; 100 mL of 5% EtOAc + 95% n-heptane; 100 mL of 10% EtOAc + 90% n-heptane; 100 mL of 20% EtOAc + 80% n-heptane; and 300 mL of 30% EtOAc + 70% n-heptane) to give the pure product (330 mg, 59% yield). 1H NMR (400 MHz, CDCl3): δ 5.22-5.19 (m, 1H), 4.41-4.38 (m, 1H), 4.11-4.07 (m, 5H), 2.47-2.38 (m, 6H), 2.34-2.26 (m, 8H), 1.99-1.92 (m, 4H), 1.62-1.53 ​​(m, 4H), 1.46-1.36 (m, 4H), 1.32-1.20 (m, 42H), 0.88-0.85 (m, 12H).

[0130] Synthesis of compound 31 [ka]

[0131] Step 1: Synthesis of compound 31-2 2-Hexyldecanoic acid (31-1, 3.6 g, 14 mmol), DCC (3.2 g, 15.4 mmol), DMAP (2.1 g, 16.9 mmol), and DCM (15 mL) were added to a 50 mL flask, and the mixture was stirred at room temperature for 30 min. Then, 1,6-hexanediol (4.98 g, 42.1 mmol) was added dropwise, and the mixture was stirred at room temperature overnight. The extent of the reaction was monitored by TLC (50% EtOAc / n-heptane). After completion of the reaction, the mixture was filtered, and the filtrate was washed with 10 mL of water and 10 mL of saturated brine, dried over anhydrous Na2SO4, and filtered. The organic phase was concentrated to give the crude product (approximately 6 g). The crude product was purified and separated by flash column (100 g silica gel; mobile phases were 100 mL each of n-heptane; 200 mL of 1% EtOAc + 99% n-heptane; 200 mL of 2% EtOAc + 98% n-heptane; 200 mL of 8% EtOAc + 92% n-heptane; and 1000 mL of 20% EtOAc + 80% n-heptane) to give the pure product (3 g, 60% yield).

[0132] Step 2: Synthesis of compound 31-3 31-2 (1.0 g, 2.8 mmol), triethylamine (851.3 mg, 8.4 mmol), and DCM (10 mL) were added to a 50 mL flask, and the mixture was cooled to 0–5 °C in an ice bath. Methanesulfonic anhydride (732.7 mg, 4.2 mmol) was added dropwise while maintaining the temperature at 0–5 °C, and the mixture was stirred at room temperature overnight. The extent of the reaction was monitored by TLC (30% EtOAc / n-heptane). After completion of the reaction, the reaction was quenched with 2 mL of water, and the mixture was extracted with DCM (5 mL × 3). The organic phases were combined, dried over anhydrous Na2SO4, filtered, and concentrated to give the crude product (approximately 2 g). The crude product was purified and separated by flash column (60 g of silica gel packed with 100 mL of n-heptane; the mobile phases were 100 mL of 1% EtOAc + 99% n-heptane; 100 mL of 2% EtOAc + 98% n-heptane; 100 mL of 5% EtOAc + 95% n-heptane; and 500 mL of 20% EtOAc + 80% n-heptane), to give the pure product (1.2 g, 98% yield).

[0133] Step 3: Synthesis of compound 31 DMF (1 mL) was added to a 10 mL flask, and sodium hydride (46 mg, 1.2 mmol) was added in batches. The mixture was cooled to 5-10 °C in an ice bath, and a mixture of 3-dimethylamino-1,2-propanediol (45.7 mg, 0.38 mmol) and DMF (1 mL) was added dropwise. After the addition, the mixture was heated to 50 °C and reacted for 10 min, then cooled again to 5-10 °C. Then, a mixture of 31-3 (500 mg, 1.2 mmol) and DMF (1 mL) was added dropwise, and the mixture was stirred at room temperature overnight. The degree of reaction was monitored by TLC (30% EtOAc / n-heptane). After completion of the reaction, the reaction was quenched with 1 mL of water, and the mixture was extracted with EtOAc (5 mL × 3). The organic phases were combined, dried over anhydrous Na2SO4, filtered, and concentrated to give the crude product (approximately 200 mg). The crude product was purified and separated by flash column (50 g silica gel; mobile phases were 100 mL of n-heptane; 100 mL of 1% EtOAc+99% n-heptane; 100 mL of 2% EtOAc+98% n-heptane; 100 mL of 5% EtOAc+95% n-heptane; 100 mL of 10% EtOAc+90% n-heptane; and 300 mL of 20% EtOAc+80% n-heptane) to give the pure product (40 mg, 13% yield).

[0134] Synthesis of compound 32 [ka]

[0135] Step 1: Synthesis of compound 32-2 Suberic acid (5.72 g, 32.8 mmol), DCC (6.77 g, 32.8 mmol), DMAP (802.3 mg, 6.6 mmol), and DCM (80 mL) were added to a 250 mL flask, and the mixture was stirred at room temperature for 30 min. Then, a solution of pentadecan-7-ol (32-1, 1.5 g, 6.6 mmol) in DCM (40 mL) was added dropwise, and the mixture was stirred at room temperature for 5 h. The extent of the reaction was monitored by TLC (30% EtOAc / n-heptane). After completion of the reaction, the mixture was filtered, and the filtrate was washed with 15 mL of water and 15 mL of saturated brine, dried over anhydrous Na2SO4, and filtered. The organic phase was concentrated to give the crude product (approximately 3.7 g). The crude product was purified and separated by flash column (50 g silica gel; mobile phases were 200 mL n-heptane; 150 mL 1% EtOAc + 99% n-heptane; 150 mL 2% EtOAc + 98% n-heptane; 200 mL 8% EtOAc + 92% n-heptane; and 1000 mL 20% EtOAc + 80% n-heptane) to give the pure product (2.1 g, 83% yield).

[0136] Step 2: Synthesis of compound 32 DMAP (39.7 mg, 0.33 mmol), DCC (268.2 mg, 1.3 mmol), and DCM (1.5 mL) were added to a 25 mL flask and stirred until the mixture became clear. 32-2 (500 mg, 1.3 mmol) was added, and the mixture was stirred at room temperature for 30 min. 3-Dimethylamino-1,2-propanediol (77.5 mg, 0.65 mmol) was then added in one batch, and the mixture was stirred at room temperature for 5 h. The extent of the reaction was monitored by TLC (5% MeOH / DCM). After completion of the reaction, the reaction solution was filtered to remove solids. The filtrate was washed with 5 mL of water and 5 mL of saturated brine, dried over anhydrous Na2SO4, and filtered. The organic phase was concentrated to give the crude product (approximately 510 mg). The crude product was purified and separated by flash column (20 g silica gel packed with 100 mL of n-heptane; mobile phases were 100 mL of 1% EtOAc + 99% n-heptane; 100 mL of 2% EtOAc + 98% n-heptane; 100 mL of 3% EtOAc + 97% n-heptane; 100 mL of 5% EtOAc + 95% n-heptane; 100 mL of 10% EtOAc + 90% n-heptane; and 100 mL of 20% EtOAc + 80% n-heptane) to give the pure product (260 mg, 47% yield).

[0137] Synthesis of compound 33 [ka]

[0138] Step 1: Synthesis of compound 33-2 Sebacic acid (6.64 g, 32.8 mmol), DCC (6.77 g, 32.8 mmol), DMAP (802.3 mg, 6.6 mmol), and DCM (80 mL) were added to a 250 mL flask, and the mixture was stirred at room temperature for 30 min. Then, a solution of pentadecan-7-ol (33-1, 1.5 g, 6.6 mmol) in DCM (40 mL) was added dropwise, and the mixture was stirred at room temperature for 5 h. The extent of the reaction was monitored by TLC (30% EtOAc / n-heptane). After completion of the reaction, the mixture was filtered, and the filtrate was washed with 15 mL of water and 15 mL of saturated brine, dried over anhydrous Na2SO4, and filtered. The organic phase was concentrated to give the crude product (approximately 3.7 g). The crude product was purified and separated by flash column (50 g silica gel; mobile phases were 200 mL n-heptane; 150 mL 1% EtOAc + 99% n-heptane; 150 mL 2% EtOAc + 98% n-heptane; 200 mL 8% EtOAc + 92% n-heptane; and 1000 mL 20% EtOAc + 80% n-heptane) to give the pure product (2.1 g, 77% yield). 1 H NMR (400 MHz, CDCl3): δ 4.89-4.85 (m, 1H), 2.36-2.25 (m, 4H), 1.66-1.47 (m, 8H), 1.37-1.25 (m, 30H), 0.89-0.85 (m, 6H).

[0139] Step 2: Synthesis of compound 33 DMAP (74 mg, 0.61 mmol), DCC (250 mg, 1.21 mmol), and DCM (1.5 mL) were added to a 25 mL flask and stirred until the mixture became clear. 33-2 (500 mg, 1.21 mmol) was added, and the mixture was stirred at room temperature for 30 min. 3-Dimethylamino-1,2-propanediol (72.2 mg, 0.61 mmol) was then added in one batch, and the mixture was stirred at room temperature for 5 h. The extent of the reaction was monitored by TLC (10% MeOH / DCM). After completion of the reaction, the reaction solution was filtered to remove solids. The filtrate was washed with 5 mL of water and 5 mL of saturated brine, dried over anhydrous Na2SO4, and filtered. The organic phase was concentrated to give the crude product (approximately 500 mg). The crude product was purified and separated by flash column (20 g silica gel packed with 100 mL of n-heptane; the mobile phases were 100 mL of 1% EtOAc + 99% n-heptane; 100 mL of 2% EtOAc + 98% n-heptane; 100 mL of 3% EtOAc + 97% n-heptane; 100 mL of 5% EtOAc + 95% n-heptane; 100 mL of 10% EtOAc + 90% n-heptane; 100 mL of 20% EtOAc + 80% n-heptane; and 300 mL of 30% EtOAc + 70% n-heptane), to give the pure product (230 mg, 42% yield). 1 H NMR (400 MHz, CDCl3): δ 5.23-5.17 (m, 1H), 4.89-4.83 (m, 2H), 4.37-4.33 (m, 1H), 4.11-4.06 (m, 1H), 2.51-2.45 (m, 2H), 2.37-2.25 (m, 14H), 1.64-1.47 (m, 20H), 1.37-1.25 (m, 65H), 0.89-0.85 (m, 12H).

[0140] Synthesis of compound 34 [ka]

[0141] Step 1: Synthesis of compound 34 DMAP (34.3 mg, 0.28 mmol), DCC (115.7 mg, 0.56 mmol), and DCM (1.5 mL) were added to a 50 mL flask and stirred until the mixture became clear. 34-1 (200 mg, 0.56 mmol) was added, and the mixture was stirred at room temperature for 30 min. 3-Dimethylamino-1,2-propanediol (72.2 mg, 0.61 mmol) was then added in one batch, and the mixture was stirred at room temperature for 5 h. The extent of the reaction was monitored by TLC (10% MeOH / DCM). After completion of the reaction, the reaction solution was filtered to remove solids. The filtrate was washed with 5 mL of water and 5 mL of saturated brine, dried over anhydrous Na2SO4, and filtered. The organic phase was concentrated to give the crude product (approximately 300 mg). The crude product was purified and separated by flash column (20 g silica gel packed with 100 mL of n-heptane; the mobile phases were 100 mL of 1% EtOAc + 99% n-heptane; 100 mL of 2% EtOAc + 98% n-heptane; 100 mL of 3% EtOAc + 97% n-heptane; 100 mL of 10% EtOAc + 90% n-heptane; 100 mL of 30% EtOAc + 70% n-heptane; 100 mL of 50% EtOAc + 50% n-heptane; and 300 mL of 80% EtOAc + 20% n-heptane) to give the pure product (60 mg, 27% yield). 1 H NMR (400 MHz, CDCl3): δ 6.53-6.50 (m, 1H), 5.05-5.00 (m, 1H), 4.89-4.81 (m, 2H), 3.56-3.40 (m, 2H), 2.45-2.37 (m, 3H), 2.36-2.28 (m, 6H), 2.25 (s, 6H), 2.21-2.17 (m, 2H), 1.69-1.61 (m, 8H), 1.52-1.47 (m, 8H), 1.33-1.25 (m, 43H), 0.89-0.85 (m, 12H).

[0142] Synthesis of compound 35 [ka]

[0143] Step 1: Synthesis of compound 35-3 35-2 (300 mg, 0.84 mmol), DCC (173.6 mg, 0.84 mmol), DMAP (51.4 mg, 0.42 mmol), and DCM (1.5 mL) were added to a 25 mL flask, and the mixture was stirred at room temperature for 30 min. 35-1 (163.1 mg, 0.42 mmol) was then added dropwise, and the mixture was stirred at room temperature overnight. The extent of the reaction was monitored by TLC (5% MeOH / DCM). After completion of the reaction, the mixture was filtered, and the filtrate was washed with 5 mL of water and 5 mL of saturated brine, dried over anhydrous Na2SO4, and filtered. The organic phase was concentrated to give the crude product (approximately 420 mg). The crude product was purified and separated by flash column (silica gel 30 g; mobile phases were 100 mL each of n-heptane; 100 mL of 1% EtOAc + 99% n-heptane; 100 mL of 2% EtOAc + 98% n-heptane; 100 mL of 8% EtOAc + 92% n-heptane; and 300 mL of 20% EtOAc + 80% n-heptane) to give the pure product (150 mg, 33% yield). 1 H NMR (400 MHz, CDCl3): δ 7.68-7.66 (m, 4H), 7.43-7.37 (m, 6H), 4.89-4.83 (m, 2H), 4.16-4.12 (m, 1H), 4.01-3.97 (m, 1H), 3.90-3.84 (m, 1H), 3.78-3.68 (m, 2H), 2.75-2.69 (m, 1H), 2.63-2.45 (m, 3H), 2.39-2.26 (m, 7H), 1.67-1.64 (m, 5H), 1.51-1.49 (m, 4H), 1.29-1.25 (m, 22H), 1.05-1.03 (m, 8H), 0.89-0.86 (m, 6H).

[0144] Step 2: Synthesis of compound 35 35-3 (140 mg, 0.13 mmol) and THF (2 mL) were added to a 25 mL flask, and the mixture was cooled to 0–5 °C in an ice bath. TBAF (103.2 mg, 0.39 mmol) was added dropwise while maintaining the temperature at 0–5 °C, and the mixture was stirred at this temperature for 3 h. The extent of the reaction was monitored by TLC (5% MeOH / DCM). After completion of the reaction, the reaction was quenched with saturated aqueous ammonium chloride solution, and the mixture was extracted with EtOAc (5 mL × 2). The organic phases were combined, dried over anhydrous Na2SO4, filtered, and concentrated to give the crude product (approximately 510 mg). The crude product was purified and separated by flash column (20 g silica gel packed with 100 mL of n-heptane; mobile phases were 100 mL of 1% EtOAc+99% n-heptane; 100 mL of 2% EtOAc+98% n-heptane; 100 mL of 3% EtOAc+97% n-heptane; 100 mL of 5% EtOAc+95% n-heptane; 100 mL of 10% EtOAc+90% n-heptane; and 300 mL of 30% EtOAc+70% n-heptane), to give the pure product (80 mg, 74% yield). 1 H NMR (400 MHz, CDCl3): δ 5.22-5.20 (m, 1H), 4.88-4.82 (m, 2H), 4.36-4.32 (m, 1H), 4.13-4.08 (m, 1H), 3.64-3.47 (m, 2H), 2.65-2.57 (m, 4H), 2.37-2.28 (m, 11H), 1.69-1.64 (m, 8H), 1.52-1.25 (m, 53H), 0.89-0.85 (m, 12H).

[0145] Synthesis of compound 36 [ka]

[0146] Step 1: Synthesis of compound 36-2 36-1 (480 mg, 1.46 mmol), DCC (301.5 mg, 1.46 mmol), DMAP (89.3 mg, 0.73 mmol), and DCM (3 mL) were added to a 25 mL flask, and the mixture was stirred at room temperature for 30 min. 35-1 (283.2 mg, 0.73 mmol) was then added dropwise, and the mixture was stirred at room temperature overnight. The extent of the reaction was monitored by TLC (30% EtOAc / n-heptane). After completion of the reaction, the mixture was filtered, and the filtrate was washed with 5 mL of water and 5 mL of saturated brine, dried over anhydrous Na2SO4, and filtered. The organic phase was concentrated to give the crude product (approximately 420 mg). The crude product was purified and separated by flash column (30 g silica gel; mobile phases were 100 mL of n-heptane; 100 mL of 1% EtOAc + 99% n-heptane; 100 mL of 2% EtOAc + 98% n-heptane; 100 mL of 8% EtOAc + 92% n-heptane; and 500 mL of 12% EtOAc + 88% n-heptane) to give the pure product (450 mg, 31% yield). 1 H NMR (400 MHz, CDCl3): δ 7.68-7.62 (m, 4H), 7.44-7.36 (m, 6H), 5.16-5.10 (m, 1H), 4.90-4.83 (m, 2H), 4.37-4.33 (m, 1H), 4.14-4.10 (m, 1H), 3.58 (t, J=4Hz, 2H), 2.63-2.55 (m, 11H), 2.25 (s, 3H), 1.53-1.48 (m, 8H), 1.37-1.19 (m, 43H), 1.04 (s, 9H), 0.89-0.86 (m, 12H).

[0147] Step 2: Synthesis of compound 36 36-2 (480 mg, 0.46 mmol) and THF (4 mL) were added to a 25 mL flask, and the mixture was cooled to 0–5 °C in an ice bath. TBAF (363.2 mg, 1.39 mmol) was added dropwise while maintaining the temperature at 0–5 °C, and the mixture was stirred at this temperature for 3 h. The extent of the reaction was monitored by TLC (5% MeOH / DCM). After completion of the reaction, the reaction was quenched with saturated aqueous ammonium chloride solution, and the mixture was extracted with EtOAc (5 mL × 2). The organic phases were combined, dried over anhydrous Na2SO4, filtered, and concentrated to give the crude product (approximately 500 mg). The crude product was purified and separated by flash column (20 g silica gel packed with 100 mL of n-heptane; mobile phases were 100 mL of 1% EtOAc + 99% n-heptane; 100 mL of 2% EtOAc + 98% n-heptane; 100 mL of 5% EtOAc + 95% n-heptane; 100 mL of 10% EtOAc + 90% n-heptane; 100 mL of 20% EtOAc + 80% n-heptane; and 300 mL of 50% EtOAc + 50% n-heptane) to give the pure product (210 mg, 57% yield). 1 H NMR (400 MHz, CDCl3): δ 5.26-5.20 (m, 1H), 4.89-4.83 (m, 2H), 4.34-4.31 (m, 1H), 4.19-4.15 (m, 1H), 3.58 (t, J=4Hz, 2H), 2.69-2.56 (m, 12H), 2.33 (s, 3H), 1.53-1.48 (m, 8H), 1.37-1.25 (m, 41H), 0.89-0.86 (m, 12H).

[0148] Synthesis of compound 37 [ka]

[0149] Step 1: Synthesis of compound 37-2 37-1 (500 mg, 1.46 mmol), DCC (301.5 mg, 1.46 mmol), DMAP (89.3 mg, 0.73 mmol), and DCM (3 mL) were added to a 25 mL flask, and the mixture was stirred at room temperature for 30 min. 35-1 (283 mg, 0.73 mmol) was then added dropwise, and the mixture was stirred at room temperature overnight. The extent of the reaction was monitored by TLC (30% EtOAc / n-heptane). After completion of the reaction, the mixture was filtered, and the filtrate was washed with 5 mL of water and 5 mL of saturated brine, dried over anhydrous Na2SO4, and filtered. The organic phase was concentrated to give the crude product (approximately 820 mg). The crude product was purified and separated by flash column (silica gel 30 g; mobile phases were 100 mL of n-heptane; 100 mL of 1% EtOAc + 99% n-heptane; 100 mL of 2% EtOAc + 98% n-heptane; 100 mL of 8% EtOAc + 92% n-heptane; and 300 mL of 10% EtOAc + 90% n-heptane) to give the pure product (480 mg, 63% yield). 1 H NMR (400 MHz, CDCl3): δ 7.68-7.64 (m, 4H), 7.44-7.36 (m, 6H), 5.17-5.12 (m, 1H), 4.39-4.39 (m, 1H), 4.11-4.06 (m, 5H), 3.71 (t, J=4Hz, 2H), 2.65-2.55 (m, 4H), 2.38-2.26 (m, 9H), 1.97-1.90 (m, 4H), 1.62-1.53 ​​(m, 5H), 1.47-1.40 (m, 4H), 1.31-1.30 (m, 43H), 1.04 (s, 9H), 0.89-0.86 (m, 12H).

[0150] Step 2: Synthesis of compound 37 37-2 (530 mg, 0.53 mmol) and THF (4 mL) were added to a 25 mL flask, and the mixture was cooled to 0–5 °C in an ice bath. TBAF (412.2 mg, 1.58 mmol) was added dropwise while maintaining the temperature at 0–5 °C, and the mixture was stirred at this temperature for 3 h. The extent of the reaction was monitored by TLC (5% MeOH / DCM). After completion of the reaction, the reaction was quenched with saturated aqueous ammonium chloride solution, and the mixture was extracted with EtOAc (5 mL × 2). The organic phases were combined, dried over anhydrous Na2SO4, filtered, and concentrated to give the crude product (approximately 500 mg). The crude product was purified and separated by flash column (20 g silica gel packed with 100 mL of n-heptane; mobile phases were 100 mL of 1% EtOAc+99% n-heptane; 100 mL of 2% EtOAc+98% n-heptane; 100 mL of 5% EtOAc+95% n-heptane; 100 mL of 10% EtOAc+90% n-heptane; and 300 mL of 20% EtOAc+80% n-heptane), to give the pure product (190 mg, 47% yield). 1 H NMR (400 MHz, CDCl3): δ 5.24-5.22 (m, 1H), 4.39-4.35 (m, 1H), 4.15-4.08 (m, 5H), 3.58 (t, J=4Hz, 2H), 2.64-2.58 (m, 5H), 2.43-2.27 (m, 10H), 2.00-1.92 (m, 4H), 1.61-1.53 ​​(m, 5H), 1.47-1.37 (m, 5H), 1.33-1.19 (m, 45H), 0.89-0.85 (m, 12H).

[0151] Synthesis of compound 38 [ka]

[0152] Step 1: Synthesis of compound 38-2 38-1 (500 mg, 1.3 mmol), DCC (268.2 mg, 1.3 mmol), DMAP (79.4 mg, 0.65 mmol), and DCM (2 mL) were added to a 25 mL flask, and the mixture was stirred at room temperature for 30 min. Then, 35-1 (252 mg, 0.65 mmol) was added dropwise, and the mixture was stirred at room temperature overnight. The extent of the reaction was monitored by TLC (5% MeOH / DCM). After completion of the reaction, the mixture was filtered, and the filtrate was washed with 5 mL of water and 5 mL of saturated brine, dried over anhydrous Na2SO4, and filtered. The organic phase was concentrated to give the crude product (approximately 1.7 g). The crude product was purified and separated by flash column (50 g silica gel; mobile phases were 200 mL n-heptane; 150 mL 1% EtOAc+99% n-heptane; 150 mL 2% EtOAc+98% n-heptane; 150 mL 8% EtOAc+92% n-heptane; 150 mL 20% EtOAc+80% n-heptane; and 500 mL 30% EtOAc+70% n-heptane) to give the pure product (700 mg, 96% yield). 1 H NMR (400 MHz, CDCl3): δ 7.68-7.65 (m, 4H), 7.42-7.36 (m, 6H), 5.12 (s, 1H), 4.87-4.84 (m, 2H), 4.35-4.31 (m, 1H), 4.09-4.04 (m, 1H), 3.72-3.69 (m, 2H), 2.61-2.54 (m, 4H), 2.30-2.24 (m, 11H), 1.65-1.48 (m, 18H), 1.37-119 (m, 53H), 1.04 (s, 9H), 0.89-0.86 (m, 12H).

[0153] Step 2: Synthesis of compound 38 38-2 (700 mg, 0.63 mmol) and THF (5 mL) were added to a 50 mL flask, and the mixture was cooled to 0–5 °C in an ice bath. TBAF (496.1 mg, 1.9 mmol) was added dropwise while maintaining the temperature at 0–5 °C, and the mixture was stirred at this temperature for 3 h. The extent of the reaction was monitored by TLC (5% MeOH / DCM). After completion of the reaction, the reaction was quenched with saturated aqueous ammonium chloride solution, and the mixture was extracted with EtOAc (5 mL × 2). The organic phases were combined, dried over anhydrous Na2SO4, filtered, and concentrated to give the crude product (approximately 510 mg). The crude product was purified and separated by flash column (20 g silica gel packed with 100 mL of n-heptane; mobile phases were 100 mL of 1% EtOAc + 99% n-heptane; 100 mL of 2% EtOAc + 98% n-heptane; 100 mL of 3% EtOAc + 97% n-heptane; 100 mL of 5% EtOAc + 95% n-heptane; 100 mL of 10% EtOAc + 90% n-heptane; and 500 mL of 30% EtOAc + 70% n-heptane) to give the pure product (300 mg, 54% yield). 1 H NMR (400 MHz, CDCl3): δ 5.21-5.19 (m, 1H), 4.86-4.83 (m, 2H), 4.34-4.30 (m, 1H), 4.11-4.07 (m, 1H), 3.57 (t, J=4Hz, 2H), 2.70-2.56 (m, 5H), 2.32-2.24 (m, 12H), 1.64-1.57 (m, 9H), 1.51-1.46 (m, 9H), 1.37-1.24 (m, 53H), 0.88-0.84 (m, 12H).

[0154] Synthesis of compound 43 [ka]

[0155] Step 1: Synthesis of compound 43-3 Sebacic acid (43-2, 6.79 g, 39 mmol), DCC (8.04 g, 39 mmol), DMAP (0.95 g, 7.8 mmol), and DCM (150 mL) were added to a 250 mL flask, and the mixture was stirred at room temperature for 30 min. Then, 9-heptadecanol (43-1, 2.0 g, 7.8 mmol) was added dropwise, and the mixture was stirred at room temperature overnight. The extent of the reaction was monitored by TLC (50% EtOAc / n-heptane). After completion of the reaction, the mixture was filtered, and the filtrate was washed with 30 mL of water and 30 mL of saturated brine, dried over anhydrous Na2SO4, and filtered. The organic phase was concentrated to give the crude product (approximately 6 g). The crude product was purified and separated by flash column (100 g silica gel; mobile phases were 100 mL each of n-heptane; 200 mL of 1% EtOAc + 99% n-heptane; 200 mL of 2% EtOAc + 98% n-heptane; 200 mL of 8% EtOAc + 92% n-heptane; 200 mL of 20% EtOAc + 80% n-heptane; and 1000 mL of 50% EtOAc + 50% n-heptane) to give the pure product (2.6 g, 81% yield).

[0156] Step 2: Synthesis of compound 43-4 43-3 (500 mg, 1.2 mmol), DCC (250 mg, 1.2 mmol), DMAP (74 mg, 0.6 mmol), and DCM (3 mL) were added to a 25 mL flask, and the mixture was stirred at room temperature for 30 min. Then, B048n-4 (234.8 mg, 0.6 mmol) was added dropwise, and the mixture was stirred at room temperature overnight. The extent of the reaction was monitored by TLC (5% MeOH / DCM). After completion of the reaction, the mixture was filtered, and the filtrate was washed with 5 mL of water and 5 mL of saturated brine, dried over anhydrous Na2SO4, and filtered. The organic phase was concentrated to give the crude product (approximately 900 mg). The crude product was purified and separated by flash column (50 g silica gel; mobile phases were 100 mL of n-heptane; 100 mL of 1% EtOAc + 99% n-heptane; 100 mL of 2% EtOAc + 98% n-heptane; and 300 mL of 5% EtOAc + 95% n-heptane) to give the pure product (430 mg, 60% yield). 1H NMR (400 MHz, CDCl3): δ 7.68-7.65 (m, 4H), 7.42-7.36 (m, 6H), 5.13-5.12 (m, 1H), 4.88-4.85 (m, 2H), 4.34-4.30 (m, 1H), 4.10-4.05 (m, 1H), 3.71 (t, J=4Hz, 2H), 2.63-2.55 (m, 4H), 2.30-2.24 (m, 11H), 1.64-1.48 (m, 18H), 1.31-1.19 (m, 60H), 1.04 (s, 9H), 0.89-0.86 (m, 12H).

[0157] Step 3: Synthesis of compound 43 43-4 (440 mg, 0.37 mmol) and THF (3 mL) were added to a 25 mL flask, and the mixture was cooled to 0–5 °C in an ice bath. TBAF (293.3 mg, 1.12 mmol) was added dropwise while maintaining the temperature at 0–5 °C, and the mixture was stirred at this temperature for 3 h. The extent of the reaction was monitored by TLC (5% MeOH / DCM). After completion of the reaction, the reaction was quenched with saturated aqueous ammonium chloride solution, and the mixture was extracted with EtOAc (5 mL × 2). The organic phases were combined, dried over anhydrous Na2SO4, filtered, and concentrated to give the crude product (approximately 500 mg). The crude product was purified and separated by flash column (50 g of silica gel packed with 100 mL of n-heptane; the mobile phases were 100 mL of 1% EtOAc+99% n-heptane; 100 mL of 2% EtOAc+98% n-heptane; 100 mL of 5% EtOAc+95% n-heptane; 100 mL of 20% EtOAc+80% n-heptane; and 400 mL of 50% EtOAc+50% n-heptane), to give the pure product (280 mg, 80% yield). 1H NMR (400 MHz, CDCl3): δ 5.22-5.19 (m, 1H), 4.89-4.83 (m, 2H), 4.34-4.31 (m, 1H), 4.12-4.08 (m, 1H), 3.58 (t, J=4Hz, 2H), 2.65-2.58 (m, 4H), 2.33-2.25 (m, 11H), 1.64-1.45 (m, 16H), 1.36-1.25 (m, 57H), 0.89-0.85 (m, 12H).

[0158] Synthesis of compound 44 [ka]

[0159] Step 1: Synthesis of compound 44 DMAP (65.9 mg, 0.54 mmol), DCC (222.7 mg, 1.1 mmol), and DCM (1.5 mL) were added to a 25 mL flask and stirred until the mixture became clear. 44-2 (400 mg, 1.1 mmol) was added, and the mixture was stirred at room temperature for 30 min. Then, 1-amino-3-(dimethylamino)propanol (44-1, 63.8 mg, 0.55 mmol) was added in one batch, and the mixture was stirred at room temperature overnight. The extent of the reaction was monitored by TLC (10% MeOH / DCM). After completion of the reaction, the reaction solution was filtered to remove solids. The filtrate was washed with 5 mL of water and 5 mL of saturated brine, dried over anhydrous Na2SO4, and filtered. The organic phase was concentrated to give the crude product (approximately 510 mg). The crude product was purified and separated by flash column (20 g silica gel packed with 100 mL of n-heptane; mobile phases were 100 mL of 1% EtOAc + 99% n-heptane; 100 mL of 2% EtOAc + 98% n-heptane; 100 mL of 3% EtOAc + 97% n-heptane; 100 mL of 5% EtOAc + 95% n-heptane; 100 mL of 10% EtOAc + 90% n-heptane; 100 mL of 20% EtOAc + 80% n-heptane; and 300 mL of 50% EtOAc + 50% n-heptane) to give the pure product (300 mg, 67% yield). 1H NMR (400 MHz, CDCl3): δ 6.32-6.29 (m, 1H), 5.03-4.97 (m, 2H), 4.07-4.04 (m, 4H), 3.02 (m, 2H), 2.49-2.39 (m, 2H), 2.33-2.26 (m, 11H), 2.18-2.14 (m, 2H), 1.69-1.53 ​​(m, 12H), 1.46-1.24 (m, 51H), 0.88-0.85 (m, 12H).

[0160] Synthesis of compound 45 [ka]

[0161] Step 1: Synthesis of compound 45-3 DMAP (1.54 g, 12.6 mmol), DCC (13.1 g, 63.2 mmol), and DCM (100 mL) were added to a 250 mL flask and stirred until the mixture became clear. Suberic acid (45-2, 11 g, 63.2 mmol) was added, and the mixture was stirred at room temperature for 30 min. 2-Decanol (45-1, 2 g, 12.6 mmol) was then added in one batch, and the mixture was stirred at room temperature overnight. The extent of the reaction was monitored by TLC (10% MeOH / DCM). After completion of the reaction, the reaction solution was filtered to remove solids. The filtrate was washed with 50 mL of water and 50 mL of saturated brine, dried over anhydrous Na2SO4, and filtered. The organic phase was concentrated to give the crude product (approximately 5.3 g). The crude product was purified and separated by flash column (silica gel 100, packed with 200 mL of n-heptane; mobile phases were 200 mL of 1% EtOAc+99% n-heptane; 200 mL of 2% EtOAc+98% n-heptane; 200 mL of 3% EtOAc+97% n-heptane; 200 mL of 5% EtOAc+95% n-heptane; 200 mL of 10% EtOAc+90% n-heptane; and 1000 mL of 20% EtOAc+80% n-heptane), to give the pure product (3.4 g, 85% yield).

[0162] Step 2: Synthesis of compound 45-4 45-3 (500 mg, 1.6 mmol), DCC (328.1 mg, 1.6 mmol), DMAP (97.1 mg, 0.8 mmol), and DCM (5 mL) were added to a 25 mL flask, and the mixture was stirred at room temperature for 30 min. Then, 3-dimethylamino-1,2-propanediol (189.5 mg, 1.6 mmol) was added, and the mixture was stirred at room temperature overnight. The extent of the reaction was monitored by TLC (10% MeOH / DCM). After completion of the reaction, the mixture was filtered, and the filtrate was washed with 5 mL of water and 5 mL of saturated brine, dried over anhydrous Na2SO4, and filtered. The organic phase was concentrated to give the crude product (approximately 700 mg). The crude product was purified and separated by flash column (50 g silica gel; mobile phases were 100 mL each of n-heptane; 100 mL of 1% EtOAc + 99% n-heptane; 100 mL of 2% EtOAc + 98% n-heptane; 100 mL of 8% EtOAc + 92% n-heptane; 100 mL of 20% EtOAc + 80% n-heptane; and 300 mL of 10% MeOH + 90% DCM) to give the pure product (480 mg, 73% yield).

[0163] Step 3: Synthesis of compound 45 DMAP (29.4 mg, 0.24 mmol), DCC (99.3 mg, 0.48 mmol), and DCM (1 mL) were added to a 25 mL flask and stirred until the mixture became clear. 45-5 (198.6 mg, 0.48 mmol) was added, and the mixture was stirred at room temperature for 30 min. 45-4 (200 mg, 0.48 mmol) was then added in one batch, and the mixture was stirred at room temperature overnight. The extent of the reaction was monitored by TLC (10% MeOH / DCM). After completion of the reaction, the reaction solution was filtered to remove solids, and the filtrate was washed with 5 mL of water and 5 mL of saturated brine, dried over anhydrous Na2SO4, and filtered. The organic phase was concentrated to give the crude product (approximately 500 mg). The crude product was purified and separated by flash column (20 g silica gel packed with 100 mL of n-heptane; mobile phases were 100 mL of 1% EtOAc + 99% n-heptane; 100 mL of 2% EtOAc + 98% n-heptane; 100 mL of 3% EtOAc + 97% n-heptane; 100 mL of 5% EtOAc + 95% n-heptane; 100 mL of 10% EtOAc + 90% n-heptane; 100 mL of 20% EtOAc + 80% n-heptane; and 300 mL of 50% EtOAc + 50% n-heptane) to give the pure product (110 mg, 28% yield). 1 H NMR (400 MHz, CDCl3): δ 5.21-5.18 (m, 1H), 4.91-4.84 (m, 2H), 4.37-4.34 (m, 1H), 4.10-4.06 (m, 1H), 2.47 (m, 2H), 2.33-2.24 (m, 14H), 2.33-2.26 (m, 11H), 1.65-1.41 (m, 15H), 1.37-1.18 (m, 51H), 0.89-0.85 (m, 12H).

[0164] Synthesis of compound 46 [ka]

[0165] Step 1: Synthesis of compound 46-2 46-1 (500 mg, 1.2 mmol), DCC (250 mg, 1.2 mmol), DMAP (74 mg, 0.6 mmol), and DCM (5 mL) were added to a 25 mL flask, and the mixture was stirred at room temperature for 30 min. Then, 3-dimethylamino-1,2-propanediol (144.4 mg, 1.2 mmol) was added, and the mixture was stirred at room temperature overnight. The extent of the reaction was monitored by TLC (10% MeOH / DCM). After completion of the reaction, the mixture was filtered, and the filtrate was washed with 5 mL of water and 5 mL of saturated brine, dried over anhydrous Na2SO4, and filtered. The organic phase was concentrated to give the crude product (approximately 700 mg). The crude product was purified and separated by flash column (50 g silica gel; mobile phases were 100 mL each of n-heptane; 100 mL of 1% EtOAc + 99% n-heptane; 100 mL of 2% EtOAc + 98% n-heptane; 100 mL of 8% EtOAc + 92% n-heptane; 100 mL of 20% EtOAc + 80% n-heptane; and 300 mL of 10% MeOH + 90% DCM) to give the pure product (380 mg, 61% yield).

[0166] Step 2: Synthesis of compound 46 DMAP (23.4 mg, 0.19 mmol), DCC (80.3 mg, 0.39 mmol), and DCM (1 mL) were added to a 25 mL flask and stirred until the mixture became clear. 46-3 (122.4 mg, 0.39 mmol) was added, and the mixture was stirred at room temperature for 30 min. 46-2 (200 mg, 0.39 mmol) was then added in one batch, and the mixture was stirred at room temperature overnight. The extent of the reaction was monitored by TLC (10% MeOH / DCM). After completion of the reaction, the reaction solution was filtered to remove solids, and the filtrate was washed with 5 mL of water and 5 mL of saturated brine, dried over anhydrous Na2SO4, and filtered. The organic phase was concentrated to give the crude product (approximately 500 mg). The crude product was purified and separated by flash column (20 g silica gel packed with 100 mL of n-heptane; the mobile phases were 100 mL of 1% EtOAc + 99% n-heptane; 100 mL of 2% EtOAc + 98% n-heptane; 100 mL of 3% EtOAc + 97% n-heptane; 100 mL of 5% EtOAc + 95% n-heptane; 100 mL of 10% EtOAc + 90% n-heptane; 100 mL of 20% EtOAc + 80% n-heptane; and 300 mL of 50% EtOAc + 50% n-heptane), to give the pure product (150 mg, 48% yield). 1 H NMR (400 MHz, CDCl3): δ 5.21-5.19 (m, 1H), 4.91-4.84 (m, 2H), 4.38-4.34 (m, 1H), 4.10-4.06 (m, 1H), 2.47 (m, 2H), 2.33-2.24 (m, 14H), 1.65-1.41 (m, 16H), 1.38-1.18 (m, 53H), 0.89-0.86 (m, 9H).

[0167] Synthesis of compound 47 [ka]

[0168] Step 1: Synthesis of compound 47-3 47-1 (405.7 mg, 1.3 mmol), DCC (266.2 mg, 1.3 mmol), DMAP (78.9 mg, 0.65 mmol), and DCM (5 mL) were added to a 25 mL flask, and the mixture was stirred at room temperature for 30 min. 47-2 (250 mg, 0.65 mmol) was then added dropwise, and the mixture was stirred at room temperature overnight. The extent of the reaction was monitored by TLC (5% MeOH / DCM). After completion of the reaction, the mixture was filtered, and the filtrate was washed with 5 mL of water and 5 mL of saturated brine, dried over anhydrous Na2SO4, and filtered. The organic phase was concentrated to give the crude product (approximately 900 mg). The crude product was purified and separated by flash column (50 g silica gel; mobile phases were 100 mL of n-heptane; 100 mL of 1% EtOAc+99% n-heptane; 100 mL of 2% EtOAc+98% n-heptane; 100 mL of 8% EtOAc+92% n-heptane; and 500 mL of 10% EtOAc+90% n-heptane) to give the pure product (480 mg, 76% yield). 1 H NMR (400 MHz, CDCl3): δ 7.68-5.65 (m, 4H), 7.42-7.38 (m, 6H), 5.12 (m, 1H), 4.91-4.86 (m, 2H), 4.35-4.31 (m, 1H), 4.09-4.05 (m, 1H), 3.72-3.69 (m, 2H), 2.61-2.54 (m, 5H), 2.28-2.23 (m, 11H), 1.62-1.45 (m, 14H), 1.33-1.18 (m, 42H), 1.04 (s, 9H), 0.89-0.86 (m, 6H).

[0169] Step 2: Synthesis of compound 47 47-3 (480 mg, 0.49 mmol) and THF (2 mL) were added to a 25 mL flask, and the mixture was cooled to 0–5 °C in an ice bath. TBAF (384 mg, 1.47 mmol) was added dropwise while maintaining the temperature at 0–5 °C, and the mixture was stirred at this temperature for 3 h. The extent of the reaction was monitored by TLC (5% MeOH / DCM). After completion of the reaction, the reaction was quenched with saturated aqueous ammonium chloride solution, and the mixture was extracted with EtOAc (5 mL × 2). The organic phases were combined, dried over anhydrous Na2SO4, filtered, and concentrated to give the crude product (approximately 500 mg). The crude product was purified and separated by flash column (60 g silica gel packed with 100 mL of n-heptane; mobile phases were 100 mL of 1% EtOAc + 99% n-heptane; 100 mL of 2% EtOAc + 98% n-heptane; 100 mL of 5% EtOAc + 95% n-heptane; 100 mL of 10% EtOAc + 90% n-heptane; 100 mL of 40% EtOAc + 60% n-heptane; and 300 mL of 60% EtOAc + 40% n-heptane) to give the pure product (240 mg, 66% yield). 1 H NMR (400 MHz, CDCl3): δ 5.23-5.17 (m, 1H), 4.92-4.84 (m, 2H), 4.34-4.30 (m, 1H), 4.11-4.02 (m, 1H), 3.58 (t, J=4Hz, 2H), 2.65-2.55 (m, 5H), 2.32-2.23 (m, 11H), 1.64-1.51 (m, 10H), 1.49-1.40 (m, 2H), 1.37-1.17 (m, 38H), 0.88-0.86 (m, 6H).

[0170] Synthesis of compound 48 [ka]

[0171] Step 1: Synthesis of compound 48-3 48-1 (425.9 mg, 1.03 mmol), DCC (212.9 mg, 1.03 mmol), DMAP (63 mg, 0.52 mmol), and DCM (2 mL) were added to a 25 mL flask, and the mixture was stirred at room temperature for 30 min. 48-2 (200 mg, 0.52 mmol) was then added dropwise, and the mixture was stirred at room temperature overnight. The extent of the reaction was monitored by TLC (5% MeOH / DCM). After completion of the reaction, the mixture was filtered, and the filtrate was washed with 5 mL of water and 5 mL of saturated brine, dried over anhydrous Na2SO4, and filtered. The organic phase was concentrated to give the crude product (approximately 880 mg). The crude product was purified and separated by flash column (30 g silica gel; mobile phases were 100 mL of n-heptane; 100 mL of 1% EtOAc + 99% n-heptane; 100 mL of 2% EtOAc + 98% n-heptane; 100 mL of 8% EtOAc + 92% n-heptane; and 500 mL of 10% EtOAc + 90% n-heptane) to give the pure product (500 mg, 82% yield). 1 H NMR (400 MHz, CDCl3): δ 7.68-5.65 (m, 4H), 7.44-7.36 (m, 6H), 5.13-5.11 (m, 1H), 4.89-4.83 (m, 2H), 4.35-4.31 (m, 1H), 4.09-4.05 (m, 1H), 3.72-3.69 (m, 2H), 2.65-2.54 (m, 4H), 2.28-2.22 (m, 12H), 1.65-1.19 (m, 84H), 1.04 (s, 9H), 0.89-0.86 (m, 12H).

[0172] Step 2: Synthesis of compound 48 D020n-1 (500 mg, 0.42 mmol) and THF (2 mL) were added to a 25 mL flask, and the mixture was cooled to 0–5 °C in an ice bath. TBAF (333.3 mg, 1.27 mmol) was added dropwise while maintaining the temperature at 0–5 °C, and the mixture was stirred at this temperature for 3 h. The extent of the reaction was monitored by TLC (5% MeOH / DCM). After completion of the reaction, the reaction was quenched with saturated aqueous ammonium chloride solution, and the mixture was extracted with EtOAc (5 mL × 2). The organic phases were combined, dried over anhydrous Na2SO4, filtered, and concentrated to give the crude product (approximately 600 mg). The crude product was purified and separated by flash column (20 g silica gel packed with 100 mL of n-heptane; mobile phases were 100 mL of 1% EtOAc+99% n-heptane; 100 mL of 2% EtOAc+98% n-heptane; 100 mL of 5% EtOAc+95% n-heptane; 100 mL of 10% EtOAc+90% n-heptane; and 300 mL of 40% EtOAc+60% n-heptane), to give the pure product (260 mg, 65% yield). 1 H NMR (400 MHz, CDCl3): δ 5.24-5.19 (m, 1H), 4.88-4.82 (m, 2H), 4.35-4.31 (m, 1H), 4.12-4.08 (m, 1H), 3.58 (t, J=4Hz, 2H), 2.67-2.57 (m, 5H), 2.33-2.25 (m, 11H), 1.65-1.58 (m, 8H), 1.52-1.47 (m, 8H), 1.36-1.25 (m, 56H), 0.89-0.85 (m, 12H).

[0173] Synthesis of compound 49 [ka]

[0174] Step 1: Synthesis of compound 49-3 49-1 (500 mg, 1.6 mmol), DCC (328 mg, 1.6 mmol), DMAP (97 mg, 0.8 mmol), and DCM (5 mL) were added to a 25 mL flask, and the mixture was stirred at room temperature for 30 min. 49-2 (677.9 mg, 1.75 mmol) was then added dropwise, and the mixture was stirred at room temperature overnight. The extent of the reaction was monitored by TLC (5% MeOH / DCM). After completion of the reaction, the mixture was filtered, and the filtrate was washed with 5 mL of water and 5 mL of saturated brine, dried over anhydrous Na2SO4, and filtered. The organic phase was concentrated to give the crude product (approximately 900 mg). The crude product was purified and separated by flash column (50 g silica gel; mobile phases were 100 mL of n-heptane; 100 mL of 1% EtOAc + 99% n-heptane; 100 mL of 2% EtOAc + 98% n-heptane; 100 mL of 8% EtOAc + 92% n-heptane; and 500 mL of 10% EtOAc + 90% n-heptane) to give the pure product (540 mg, 49% yield). 1 H NMR (400 MHz, CDCl3): δ 7.68-7.65 (m, 4H), 7.45-7.37 (m, 6H), 4.93-4.85 (m, 1H), 4.16-4.12 (m, 1H), 4.01-3.97 (m, 1H), 3.90-3.84 (m, 1H), 2.79-2.69 (m, 2H), 2.75-2.69 (m, 1H), 2.63-2.55 (m, 1H), 2.48-2.46 (m, 1H), 2.36-2.32 (m, 5H), 2.28-2.24 (m, 2H), 1.65-1.58 (m, 5H), 1.48-1.41 (m, 1H), 1.35-1.26 (m, 20H), 1.20-1.18 (m, 3H), 1.05 (s, 9H), 0.90-0.86 (m, 6H).

[0175] Step 2: Synthesis of compound 49-5 49-4 (325.8 mg, 0.79 mmol), DCC (162.9 mg, 0.79 mmol), DMAP (96.4 mg, 0.79 mmol), and DCM (2 mL) were added to a 25 mL flask, and the mixture was stirred at room temperature for 30 min. 49-3 (540 mg, 0.79 mmol) was then added dropwise, and the mixture was stirred at room temperature overnight. The extent of the reaction was monitored by TLC (5% MeOH / DCM). After completion of the reaction, the mixture was filtered, and the filtrate was washed with 5 mL of water and 5 mL of saturated brine, dried over anhydrous Na2SO4, and filtered. The organic phase was concentrated to give the crude product (approximately 900 mg). The crude product was purified and separated by flash column (50 g silica gel; mobile phases were 100 mL each of n-heptane; 100 mL of 1% EtOAc+99% n-heptane; 100 mL of 2% EtOAc+98% n-heptane; 100 mL of 5% EtOAc+95% n-heptane; 100 mL of 10% EtOAc+90% n-heptane; and 500 mL of 20% EtOAc+80% n-heptane) to give the pure product (650 mg, 76% yield). 1 H NMR (400 MHz, CDCl3): δ 7.68-7.65 (m, 4H), 7.42-7.36 (m, 6H), 5.14-5.11 (m, 1H), 4.91-4.84 (m, 2H), 4.35-4.31 (m, 1H), 4.09-4.05 (m, 1H), 3.72-3.69 (m, 2H), 2.63-2.55 (m, 4H), 2.30-2.24 (m, 12H), 1.64-1.42 (m, 16H), 1.37-1.18 (m, 56H), 1.04 (s, 9H), 0.90-0.86 (m, 12H).

[0176] Step 3: Synthesis of compound 49 49-5 (650 mg, 0.6 mmol) and THF (4 mL) were added to a 25 mL flask, and the mixture was cooled to 0–5 °C in an ice bath. TBAF (472.7 mg, 1.8 mmol) was added dropwise while maintaining the temperature at 0–5 °C, and the mixture was stirred at this temperature for 3 h. The extent of the reaction was monitored by TLC (5% MeOH / DCM). After completion of the reaction, the reaction was quenched with saturated aqueous ammonium chloride solution, and the mixture was extracted with EtOAc (5 mL × 2). The organic phases were combined, dried over anhydrous Na2SO4, filtered, and concentrated to give the crude product (approximately 500 mg). The crude product was purified and separated by flash column (60 g silica gel packed with 100 mL of n-heptane; mobile phases were 100 mL of 1% EtOAc+99% n-heptane; 100 mL of 2% EtOAc+98% n-heptane; 100 mL of 5% EtOAc+95% n-heptane; 100 mL of 10% EtOAc+90% n-heptane; and 500 mL of 40% EtOAc+60% n-heptane), to give the pure product (380 mg, 75% yield). 1 H NMR (400 MHz, CDCl3): δ 5.24-5.18 (m, 1H), 4.92-4.82 (m, 2H), 4.35-4.31 (m, 1H), 4.12-4.03 (m, 1H), 3.59 (t, J=4Hz, 2H), 2.68-2.58 (m, 4H), 2.37-2.24 (m, 11H), 1.65-1.14 (m, 16H), 1.35-1.18 (m, 50H), 0.89-0.85 (m, 9H).

[0177] Synthesis of compound 50 [ka]

[0178] Step 1: Synthesis of compound 50-3 50-2 (532.3 mg, 1.3 mmol), DCC (266.2 mg, 1.3 mmol), DMAP (157.6 mg, 1.3 mmol), and DCM (5 mL) were added to a 25 mL flask, and the mixture was stirred at room temperature for 30 min. Then, 50-1 (500 mg, 1.3 mmol) was added dropwise, and the mixture was stirred at room temperature overnight. The extent of the reaction was monitored by TLC (50% EtOAc / n-heptane). After completion of the reaction, the mixture was filtered, and the filtrate was washed with 5 mL of water and 5 mL of saturated brine, dried over anhydrous Na2SO4, and filtered. The organic phase was concentrated to give the crude product (approximately 900 mg). The crude product was purified and separated by flash column (50 g silica gel; mobile phases were 100 mL of n-heptane; 100 mL of 1% EtOAc+99% n-heptane; 100 mL of 2% EtOAc+98% n-heptane; 100 mL of 8% EtOAc+92% n-heptane; 100 mL of 20% EtOAc+80% n-heptane; and 400 mL of 50% EtOAc+50% n-heptane) to give the pure product (550 mg, 54% yield).

[0179] Step 2: Synthesis of compound 50-5 50-4 (221.1 mg, 0.7 mmol), DCC (145.1 mg, 0.7 mmol), DMAP (43 mg, 0.35 mmol), and DCM (5 mL) were added to a 25 mL flask, and the mixture was stirred at room temperature for 30 min. 50-3 (550 mg, 0.7 mmol) was then added dropwise, and the mixture was stirred at room temperature overnight. The extent of the reaction was monitored by TLC (5% MeOH / DCM). After completion of the reaction, the mixture was filtered, and the filtrate was washed with 5 mL of water and 5 mL of saturated brine, dried over anhydrous Na2SO4, and filtered. The organic phase was concentrated to give the crude product (approximately 900 mg). The crude product was purified and separated by flash column (50 g silica gel; mobile phases were 100 mL each of n-heptane; 100 mL of 1% EtOAc+99% n-heptane; 100 mL of 2% EtOAc+98% n-heptane; 100 mL of 5% EtOAc+95% n-heptane; 100 mL of 10% EtOAc+90% n-heptane; and 500 mL of 20% EtOAc+80% n-heptane) to give the pure product (530 mg, 70% yield). 1H NMR (400 MHz, CDCl3): δ 7.68-7.65 (m, 4H), 7.44-7.36 (m, 6H), 5.13-5.11 (m, 1H), 4.91-4.84 (m, 2H), 4.35-4.31 (m, 1H), 4.09-4.05 (m, 1H), 3.72-3.69 (m, 2H), 2.67-2.55 (m, 4H), 2.29-2.23 (m, 12H), 1.65-1.42 (m, 16H), 1.37-1.18 (m, 50H), 1.04 (s, 9H), 0.89-0.86 (m, 9H).

[0180] Step 3: Synthesis of Compound 50 50-5 (530 mg, 0.49 mmol) and THF (5 mL) were added to a 25 mL flask, and the mixture was cooled to 0–5 °C in an ice bath. TBAF (385.4 mg, 1.5 mmol) was added dropwise while maintaining the temperature at 0–5 °C, and the mixture was stirred at this temperature for 3 h. The extent of the reaction was monitored by TLC (5% MeOH / DCM). After completion of the reaction, the reaction was quenched with saturated aqueous ammonium chloride solution, and the mixture was extracted with EtOAc (5 mL × 2). The organic phases were combined, dried over anhydrous Na2SO4, filtered, and concentrated to give the crude product (approximately 530 mg). The crude product was purified and separated by flash column (60 g silica gel packed with 100 mL of n-heptane; mobile phases were 100 mL of 1% EtOAc+99% n-heptane; 100 mL of 2% EtOAc+98% n-heptane; 100 mL of 5% EtOAc+95% n-heptane; 100 mL of 10% EtOAc+90% n-heptane; and 500 mL of 30% EtOAc+70% n-heptane), to give the pure product (330 mg, 80% yield). 1H NMR (400 MHz, CDCl3): δ 5.24-5.19 (m, 1H), 4.91-4.82 (m, 2H), 4.35-4.31 (m, 1H), 4.12-4.08 (m, 1H), 3.59 (t, J=4Hz, 2H), 2.68-2.59 (m, 4H), 2.34-2.24 (m, 11H), 1.64-1.47 (m, 15H), 1.35-1.18 (m, 50H), 0.89-0.85 (m, 9H).

[0181] Synthesis of compound 51 [ka]

[0182] Step 1: Synthesis of compound 51-3 Suberic acid (51-2, 7.21 g, 41.4 mmol), DCC (8.55 g, 41.4 mmol), DMAP (1.01 g, 8.28 mmol), and DCM (100 mL) were added to a 250 mL flask, and the mixture was stirred at room temperature for 30 min. Di-n-octylamine (51-1, 2 g, 8.28 mmol) was then added dropwise, and the mixture was stirred at room temperature overnight. The extent of the reaction was monitored by TLC (5% MeOH / DCM). After completion of the reaction, the mixture was filtered, and the filtrate was washed with 15 mL of water and 15 mL of saturated brine, dried over anhydrous Na2SO4, and filtered. The organic phase was concentrated to give the crude product (approximately 4 g). The crude product was purified and separated by flash column (silica gel 120 g; mobile phases were 150 mL of n-heptane; 150 mL of 1% EtOAc + 99% n-heptane; 150 mL of 2% EtOAc + 98% n-heptane; 200 mL of 8% EtOAc + 92% n-heptane; 200 mL of 20% EtOAc + 80% n-heptane; and 1000 mL of 50% EtOAc + 50% n-heptane) to give the pure product (2.7 g, 82% yield). Step 2: Synthesis of compound 51-5 51-3 (1.7 g, 4.3 mmol), DCC (882.1 mg, 4.3 mmol), DMAP (522.3 mg, 4.3 mmol), and DCM (10 mL) were added to a 25 mL flask, and the mixture was stirred at room temperature for 30 min. 51-4 (1.66 g, 4.3 mmol) was then added dropwise, and the mixture was stirred at room temperature overnight. The extent of the reaction was monitored by TLC (5% MeOH / DCM). After completion of the reaction, the mixture was filtered, and the filtrate was washed with 5 mL of water and 5 mL of saturated brine, dried over anhydrous Na2SO4, and filtered. The organic phase was concentrated to give the crude product (approximately 2.8 g). The crude product was purified and separated by flash column (100 g silica gel; mobile phases were 100 mL each of n-heptane; 100 mL of 1% EtOAc + 99% n-heptane; 100 mL of 2% EtOAc + 98% n-heptane; 100 mL of 8% EtOAc + 92% n-heptane; 100 mL of 20% EtOAc + 80% n-heptane; and 500 mL of 50% EtOAc + 50% n-heptane) to give the pure product (1.6 g, 48% yield). Step 3: Synthesis of compound 51-7

[0183] 51-5 (259.1 mg, 0.65 mmol), DCC (134.5 mg, 0.65 mmol), DMAP (79.6 mg, 0.65 mmol), and DCM (3 mL) were added to a 25 mL flask, and the mixture was stirred at room temperature for 30 min. Then, 51-6 (500 mg, 0.65 mmol) was added dropwise, and the mixture was stirred at room temperature overnight. The extent of the reaction was monitored by TLC (5% MeOH / DCM). After completion of the reaction, the mixture was filtered, and the filtrate was washed with 5 mL of water and 5 mL of saturated brine, dried over anhydrous Na2SO4, and filtered. The organic phase was concentrated to give the crude product (approximately 800 mg). The crude product was purified and separated by flash column (50 g silica gel; mobile phases were 100 mL of n-heptane; 100 mL of 1% EtOAc+99% n-heptane; 100 mL of 2% EtOAc+98% n-heptane; 100 mL of 5% EtOAc+95% n-heptane; and 500 mL of 10% EtOAc+90% n-heptane) to give the pure product (430 mg, 58% yield).

[0184] Step 3: Synthesis of compound 51 51-7 (430 mg, 0.37 mmol) and THF (3 mL) were added to a 25 mL flask, and the mixture was cooled to 0–5 °C in an ice bath. TBAF (294.1 mg, 1.12 mmol) was added dropwise while maintaining the temperature at 0–5 °C, and the mixture was stirred at this temperature for 3 h. The extent of the reaction was monitored by TLC (5% MeOH / DCM). After completion of the reaction, the reaction was quenched with saturated aqueous ammonium chloride solution, and the mixture was extracted with EtOAc (5 mL × 2). The organic phases were combined, dried over anhydrous Na2SO4, filtered, and concentrated to give the crude product (approximately 530 mg). The crude product was purified and separated by flash column (50 g of silica gel packed with 100 mL of n-heptane; mobile phases were 100 mL of 1% EtOAc+99% n-heptane; 100 mL of 2% EtOAc+98% n-heptane; 100 mL of 5% EtOAc+95% n-heptane; 100 mL of 10% EtOAc+90% n-heptane; and 400 mL of 30% EtOAc+70% n-heptane), to give the pure product (200 mg, 80% yield). 1 H NMR (400 MHz, CDCl3): δ 5.25-5.19 (m, 1H), 4.34-4.30 (m, 1H), 4.13-4.08 (m, 1H), 3.62-3.60 (m, 2H), 3.29-3.16 (m, 8H), 2.65 (m, 4H), 2.36-2.25 (m, 11H), 1.68-1.59 (m, 8H), 1.56-1.44 (m, 8H), 1.39-1.23 (m, 52H), 0.90-0.85 (m, 12H).

[0185] Synthesis of compound 52 [ka]

[0186] Step 1: Synthesis of compound 52-3 2-Hexyldecanoic acid (52-2, 5.8 g, 22.7 mmol), DCC (4.68 g, 22.7 mmol), DMAP (2.77 g, 22.7 mmol), and DCM (100 mL) were added to a 250 mL flask, and the mixture was stirred at room temperature for 30 min. Then, 1,7-heptanediol (52-1, 3.0 g, 22.7 mmol) was added dropwise, and the mixture was stirred at room temperature overnight. The extent of the reaction was monitored by TLC (20% EtOAc / n-heptane). After completion of the reaction, the mixture was filtered, and the filtrate was washed with 100 mL of water and 100 mL of saturated brine, dried over anhydrous Na2SO4, and filtered. The organic phase was concentrated to give the crude product (approximately 9 g). The crude product was purified and separated by flash column (100 g silica gel; mobile phases were 100 mL each of n-heptane; 200 mL of 1% EtOAc + 99% n-heptane; 200 mL of 2% EtOAc + 98% n-heptane; 200 mL of 8% EtOAc + 92% n-heptane; and 1500 mL of 15% EtOAc + 85% n-heptane) to give the pure product (7 g, 83% yield).

[0187] Step 2: Synthesis of compound 52-4 52-3 (3.0 g, 8.1 mmol), DCM (60 mL), acetic acid (18 mL), and tetrabutylammonium bromide (1.3 g, 4.05 mmol) were added to a 250 mL flask, and the mixture was cooled to 0–5 °C in an ice bath. While maintaining the temperature at 0–5 °C, a mixture of potassium permanganate (3.84 g, 24.3 mmol) and water (49 mL) was added dropwise. After the addition, the mixture was stirred for 16 h. The degree of reaction was monitored by TLC (5% MeOH / DCM). After completion of the reaction, the reaction was quenched with saturated aqueous sodium sulfite solution, and the mixture was extracted with DCM (50 mL × 2). The organic phases were combined, dried over anhydrous Na2SO4, filtered, and concentrated to give the crude product (approximately 3.2 g). The crude product was purified and separated by flash column (60 g of silica gel packed with 100 mL of n-heptane; the mobile phases were 100 mL of 1% EtOAc + 99% n-heptane; 100 mL of 2% EtOAc + 98% n-heptane; and 500 mL of 5% EtOAc + 95% n-heptane), to give the pure product (2 g, 65% yield).

[0188] Step 3: Synthesis of compound 52-6 52-4 (1.0 g, 2.6 mmol), DCC (536.5 mg, 2.6 mmol), DMAP (158.8 mg, 2.6 mmol), and DCM (5 mL) were added to a 25 mL flask, and the mixture was stirred at room temperature for 30 min. 52-5 (503.9 mg, 1.3 mmol) was then added dropwise, and the mixture was stirred at room temperature overnight. The extent of the reaction was monitored by TLC (20% EtOAc / n-heptane). After completion of the reaction, the mixture was filtered, and the filtrate was washed with 5 mL of water and 5 mL of saturated brine, dried over anhydrous Na2SO4, and filtered. The organic phase was concentrated to give the crude product (approximately 2 g). The crude product was purified and separated by flash column (50 g silica gel; mobile phases were 100 mL each of n-heptane; 100 mL of 1% EtOAc+99% n-heptane; 100 mL of 2% EtOAc+98% n-heptane; 100 mL of 8% EtOAc+92% n-heptane; 100 mL of 15% EtOAc+85% n-heptane; and 400 mL of 20% EtOAc+80% n-heptane) to give the pure product (1 g, 68% yield). 1 H NMR (400 MHz, CDCl3): δ 7.68-7.65 (m, 4H), 7.45-7.37 (m, 6H), 5.04 (m, 1H), 4.14-4.10 (m, 1H), 4.07-3.98 (m, 4H), 3.96-3.90 (m, 1H), 3.79-3.72 (m, 2H), 2.81-2.51 (m, 4H), 2.38-2.27 (m, 7H), 1.67-1.53 ​​(m, 9H), 1.46-1.20 (m, 44H), 1.05 (s, 9H), 0.90-0.85 (m, 12H).

[0189] Step 3: Synthesis of Compound 52 52-6 (350 mg, 0.31 mmol) and THF (3 mL) were added to a 25 mL flask, and the mixture was cooled to 0–5 °C in an ice bath. TBAF (245 mg, 0.94 mmol) was added dropwise while maintaining the temperature at 0–5 °C, and the mixture was stirred at this temperature for 3 h. The extent of the reaction was monitored by TLC (10% MeOH / DCM). After completion of the reaction, the reaction was quenched with saturated aqueous ammonium chloride solution, and the mixture was extracted with EtOAc (5 mL × 2). The organic phases were combined, dried over anhydrous Na2SO4, filtered, and concentrated to give the crude product (approximately 530 mg). The crude product was purified and separated by flash column (50 g of silica gel packed with 100 mL of n-heptane; the mobile phases were 100 mL of 1% EtOAc+99% n-heptane; 100 mL of 2% EtOAc+98% n-heptane; 100 mL of 10% EtOAc+90% n-heptane; 100 mL of 30% EtOAc+70% n-heptane; and 400 mL of 50% EtOAc+50% n-heptane), to give the pure product (110 mg, 40% yield). 1 H NMR (400 MHz, CDCl3): δ 4.18-4.10 (m, 2H), 4.06-4.03 (m, 6H), 3.78-3.70 (m, 8H), 3.32-3.29 (m, 1H), 2.81-2.54 (m, 6H), 2.44 (s, 4H), 2.37-2.26 (m, 6H), 1.70-1.52 (m, 13H), 1.47-1.24 (m, 54H), 1.02-0.99 (m, 2H), 0.88-0.84 (m, 12H).

[0190] Synthesis of compound 53 [ka]

[0191] Step 1: Synthesis of compound 53-3 2-Hexyldecanoic acid (53-2, 2.93 g, 11.4 mmol), DCC (2.36 g, 11.4 mmol), DMAP (1.4 g, 11.4 mmol), and DCM (10 mL) were added to a 50 mL flask, and the mixture was stirred at room temperature for 30 min. 7-Aminoheptanol (53-1, 1.5 g, 11.4 mmol) was then added dropwise, and the mixture was stirred at room temperature overnight. The extent of the reaction was monitored by TLC (30% EtOAc / n-heptane). After completion of the reaction, the mixture was filtered, and the filtrate was washed with 5 mL of water and 5 mL of saturated brine, dried over anhydrous Na2SO4, and filtered. The organic phase was concentrated to give the crude product (approximately 3 g). The crude product was purified and separated by flash column (100 g silica gel; mobile phases were 100 mL each) of n-heptane; 100 mL of 1% EtOAc+99% n-heptane; 100 mL of 2% EtOAc+98% n-heptane; 100 mL of 10% EtOAc+90% n-heptane; 100 mL of 20% EtOAc+80% n-heptane; and 700 mL of 50% EtOAc+50% n-heptane) to give the pure product (1.8 g, 43% yield).

[0192] Step 2: Synthesis of compound 53-4 53-3 (2.3 g, 6 mmol), DCM (46 mL), acetic acid (14 mL), and tetrabutylammonium bromide (0.96 g, 3 mmol) were added to a 250 mL flask, and the mixture was cooled to 0–5 °C in an ice bath. While maintaining the temperature at 0–5 °C, a mixture of potassium permanganate (2.83 g, 17.9 mmol) and water (37 mL) was added dropwise. After the addition, the mixture was stirred for 16 h. The degree of reaction was monitored by TLC (5% MeOH / DCM). After completion of the reaction, the reaction was quenched with saturated aqueous sodium sulfite, and the mixture was extracted with DCM (50 mL × 2). The organic phases were combined, dried over anhydrous Na2SO4, filtered, and concentrated to give the crude product (approximately 3 g). The crude product was purified and separated by flash column (60 g of silica gel packed with 100 mL of n-heptane; the mobile phases were 100 mL of 1% EtOAc + 99% n-heptane; 100 mL of 2% EtOAc + 98% n-heptane; 100 mL of 10% EtOAc + 90% n-heptane; and 500 mL of 30% EtOAc + 70% n-heptane), to give the pure product (1.4 g, 59% yield).

[0193] Step 3: Synthesis of compound 53-6 53-4 (500 mg, 1.3 mmol), DCC (259.4 mg, 1.3 mmol), DMAP (153.6 mg, 1.3 mmol), and DCM (3 mL) were added to a 25 mL flask, and the mixture was stirred at room temperature for 30 min. 53-5 (913.1 mg, 1.3 mmol) was then added dropwise, and the mixture was stirred at room temperature overnight. The extent of the reaction was monitored by TLC (5% MeOH / DCM). After completion of the reaction, the mixture was filtered, and the filtrate was washed with 5 mL of water and 5 mL of saturated brine, dried over anhydrous Na2SO4, and filtered. The organic phase was concentrated to give the crude product (approximately 1 g). The crude product was purified and separated by flash column (30 g silica gel; mobile phases were 100 mL each of n-heptane; 100 mL of 1% EtOAc + 99% n-heptane; 100 mL of 2% EtOAc + 98% n-heptane; 100 mL of 8% EtOAc + 92% n-heptane; 100 mL of 15% EtOAc + 85% n-heptane; 100 mL of 20% EtOAc + 80% n-heptane; and 500 mL of 50% EtOAc + 50% n-heptane) to give the pure product (500 mg, 36% yield). 1 H NMR (400 MHz, CDCl3): δ 7.67-7.63 (m, 4H), 7.44-7.36 (m, 6H), 5.61-5.59 (m, 1H), 4.13-4.04 (m, 2H), 3.73-3.70 (m, 2H), 3.53-3.34 (m, 4H), 2.99-2.92 (m, 1H), 2.77-2.54 (m, 3H), 2.33-2.19 (m, 6H), 1.97-1.91 (m, 2H), 1.71-1.13 (m, 80H), 1.05 (s, 9H), 0.89-0.84 (m, 12H).

[0194] Step 3: Synthesis of compound 53 53-6 (370 mg, 0.32 mmol) and THF (3 mL) were added to a 25 mL flask, and the mixture was cooled to 0–5 °C in an ice bath. TBAF (253 mg, 0.97 mmol) was added dropwise while maintaining the temperature at 0–5 °C, and the mixture was stirred at this temperature for 3 h. The extent of the reaction was monitored by TLC (10% MeOH / DCM). After completion of the reaction, the reaction was quenched with saturated aqueous ammonium chloride solution, and the mixture was extracted with EtOAc (5 mL × 2). The organic phases were combined, dried over anhydrous Na2SO4, filtered, and concentrated to give the crude product (approximately 230 mg). The crude product was purified and separated by flash column (20 g silica gel packed with 100 mL of n-heptane; mobile phases were 100 mL of 1% EtOAc + 99% n-heptane; 100 mL of 2% EtOAc + 98% n-heptane; 100 mL of 10% EtOAc + 90% n-heptane; 100 mL of 30% EtOAc + 70% n-heptane; and 200 mL of 50% EtOAc + 50% n-heptane), to give the pure product (90 mg, 31% yield). 1 H NMR (400 MHz, CDCl3): δ 5.66-45.50 (m, 2H), 5.27-5.25 (m, 1H), 4.38-4.34 (m, 1H), 4.12-4.07 (m, 1H), 3.64-3.62 (m, 2H), 3.26-3.21 (m, 4H), 2.69 (m, 3H), 2.40-2.29 (m, 6H), 1.99-1.92 (m, 2H), 1.66-1.46 (m, 12H), 1.41-1.20(m, 56H), 0.89-0.85 (m, 12H).

[0195] Synthesis of compound 54 [ka]

[0196] Step 1: Synthesis of compound 54-3 2-Hexyldecanoic acid (54-1, 10.5 g, 41 mmol), DCC (8.47 g, 41 mmol), DMAP (5 g, 41 mmol), and DCM (120 mL) were added to a 250 mL flask, and the mixture was stirred at room temperature for 30 min. Then, 1,8-octanediol (54-2, 6.0 g, 41 mmol) was added dropwise, and the mixture was stirred at room temperature overnight. The extent of the reaction was monitored by TLC (20% EtOAc / n-heptane). After completion of the reaction, the mixture was filtered, and the filtrate was washed with 100 mL of water and 100 mL of saturated brine, dried over anhydrous Na2SO4, and filtered. The organic phase was concentrated to give the crude product (approximately 9 g). The crude product was purified and separated by flash column (100 g silica gel; mobile phases were 100 mL each of n-heptane; 200 mL of 1% EtOAc + 99% n-heptane; 200 mL of 2% EtOAc + 98% n-heptane; 200 mL of 8% EtOAc + 92% n-heptane; and 1500 mL of 15% EtOAc + 85% n-heptane) to give the pure product (7 g, 44% yield).

[0197] Step 2: Synthesis of compound 54-4 54-3 (1.0 g, 2.6 mmol), DCM (20 mL), acetic acid (6 mL), and tetrabutylammonium bromide (0.42 g, 1.3 mmol) were added to a 50 mL flask, and the mixture was cooled to 0–5 °C in an ice bath. While maintaining the temperature at 0–5 °C, a mixture of potassium permanganate (1.23 g, 7.8 mmol) and water (16 mL) was added dropwise. After the addition, the mixture was stirred for 16 h. The degree of reaction was monitored by TLC (5% MeOH / DCM). After completion of the reaction, the reaction was quenched with saturated aqueous sodium sulfite, and the mixture was extracted with DCM (50 mL × 2). The organic phases were combined, dried over anhydrous Na2SO4, filtered, and concentrated to give the crude product (approximately 1.2 g). The crude product was purified and separated by flash column (30 g silica gel packed with 100 mL of n-heptane; mobile phases were 100 mL of 1% EtOAc + 99% n-heptane; 100 mL of 2% EtOAc + 98% n-heptane; and 500 mL of 5% EtOAc + 95% n-heptane), to give the pure product (700 mg, 70% yield).

[0198] Step 3: Synthesis of compound 54-6 54-4 (1.0 g, 2.5 mmol), DCC (517.6 mg, 2.5 mmol), DMAP (153.2 mg, 1.25 mmol), and DCM (5 mL) were added to a 25 mL flask, and the mixture was stirred at room temperature for 30 min. 54-5 (486.2 mg, 1.25 mmol) was then added dropwise, and the mixture was stirred at room temperature overnight. The extent of the reaction was monitored by TLC (20% EtOAc / n-heptane). After completion of the reaction, the mixture was filtered, and the filtrate was washed with 5 mL of water and 5 mL of saturated brine, dried over anhydrous Na2SO4, and filtered. The organic phase was concentrated to give the crude product (approximately 1.8 g). The crude product was purified and separated by flash column (50 g silica gel; mobile phases were 100 mL of n-heptane; 100 mL of 1% EtOAc+99% n-heptane; 100 mL of 2% EtOAc+98% n-heptane; 100 mL of 8% EtOAc+92% n-heptane; and 300 mL of 15% EtOAc+85% n-heptane) to give the pure product (760 mg, 53% yield). 1 H NMR (400 MHz, CDCl3): δ 7.68-7.65 (m, 4H), 7.44-7.36 (m, 6H), 5.13 (m, 1H), 4.35-4.31 (m, 1H), 4.10-4.03 (m, 5H), 3.71 (m, 2H), 2.60-2.56 (m, 3H), 2.34-2.24 (m, 8H), 1.64-1.54 (m, 13H), 1.46-1.19 (m, 62H), 1.04 (s, 9H), 0.89-0.85 (m, 12H).

[0199] Step 4: Synthesis of compound 54 54-6 (750 mg, 0.65 mmol) and THF (8 mL) were added to a 50 mL flask, and the mixture was cooled to 0–5 °C in an ice bath. TBAF (512.1 mg, 1.96 mmol) was added dropwise while maintaining the temperature at 0–5 °C, and the mixture was stirred at this temperature for 3 h. The extent of the reaction was monitored by TLC (5% MeOH / DCM). After completion of the reaction, the reaction was quenched with saturated aqueous ammonium chloride solution, and the mixture was extracted with EtOAc (5 mL × 2). The organic phases were combined, dried over anhydrous Na2SO4, filtered, and concentrated to give the crude product (approximately 830 mg). The crude product was purified and separated by flash column (50 g of silica gel packed with 100 mL of n-heptane; the mobile phases were 100 mL of 1% EtOAc + 99% n-heptane; 100 mL of 2% EtOAc + 98% n-heptane; 100 mL of 10% EtOAc + 90% n-heptane; and 400 mL of 20% EtOAc + 80% n-heptane), to give the pure product (350 mg, 59% yield). 1 H NMR (400 MHz, CDCl3): δ 5.25-5.19 (m, 1H), 4.35-4.31 (m, 1H), 4.12-4.03 (m, 5H), 3.59 (t, J=4Hz, 2H), 2.69-2.58 (m, 4H), 2.35-2.26 (m, 10H), 1.63-1.53 ​​(m, 12H), 1.46-1.19 (m, 56H), 0.88-0.82 (m, 12H).

[0200] Synthesis of compound 55 [ka]

[0201] Step 1: Synthesis of compound 55-3 55-1 (232.4 mg, 0.65 mmol), DCC (134.5 mg, 0.65 mmol), DMAP (79.62 mg, 0.65 mmol), and DCM (3 mL) were added to a 25 mL flask, and the mixture was stirred at room temperature for 30 min. 55-2 (500 mg, 0.65 mmol) was then added dropwise, and the mixture was stirred at room temperature overnight. The extent of the reaction was monitored by TLC (5% MeOH / DCM). After completion of the reaction, the mixture was filtered, and the filtrate was washed with 5 mL of water and 5 mL of saturated brine, dried over anhydrous Na2SO4, and filtered. The organic phase was concentrated to give the crude product (approximately 1.2 g). The crude product was purified and separated by flash column (silica gel 60 g; mobile phases were 100 mL of n-heptane; 100 mL of 1% EtOAc + 99% n-heptane; 100 mL of 2% EtOAc + 98% n-heptane; and 500 mL of 8% EtOAc + 92% n-heptane) to give the pure product (550 mg, 76% yield). 1 H NMR (400 MHz, CDCl3): δ 7.67-7.65 (m, 4H), 7.42-7.36 (m, 6H), 5.12 (m, 1H), 4.89-4.83 (m, 1H), 4.34-4.31 (m, 1H), 4.09-4.04 (m, 1H), 3.70-3.68 (m, 2H), 3.29-3.16 (m, 5H), 2.61-2.54 (m, 3H), 2.28-2.24 (m, 10H), 1.64-1.47 (m, 20H), 1.34-1.25 (m, 52H), 1.04 (s, 9H), 0.90-0.85 (m, 12H).

[0202] Step 2: Synthesis of compound 55 55-3 (550 mg, 0.5 mmol) and THF (3 mL) were added to a 25 mL flask, and the mixture was cooled to 0–5 °C in an ice bath. TBAF (390.2 mg, 1.5 mmol) was added dropwise while maintaining the temperature at 0–5 °C, and the mixture was stirred at this temperature for 3 h. The extent of the reaction was monitored by TLC (5% MeOH / DCM). After completion of the reaction, the reaction was quenched with saturated aqueous ammonium chloride solution, and the mixture was extracted with EtOAc (5 mL × 2). The organic phases were combined, dried over anhydrous Na2SO4, filtered, and concentrated to give the crude product (approximately 530 mg). The crude product was purified and separated by flash column (50 g of silica gel packed with 100 mL of n-heptane; the mobile phases were 100 mL of 1% EtOAc+99% n-heptane; 100 mL of 2% EtOAc+98% n-heptane; 100 mL of 10% EtOAc+90% n-heptane; 100 mL of 30% EtOAc+70% n-heptane; and 400 mL of 50% EtOAc+50% n-heptane), to give the pure product (250 mg, 58% yield). 1 H NMR (400 MHz, CDCl3): δ 5.22-5.19 (m, 1H), 4.88-4.81 (m, 1H), 4.33-4.29 (m, 1H), 4.12-4.07 (m, 1H), 3.57 (t, J=4Hz, 2H), 3.28-3.16 (m, 4H), 2.66-2.56 (m, 4H), 2.35-2.24 (m, 10H), 1.65-1.44 (m, 15H), 1.36-1.24 (m, 48H), 0.89-0.84 (m, 12H).

[0203] Synthesis of compound 56 [ka]

[0204] Step 1: Synthesis of compound 56-3 56-1 (200 mg, 0.52 mmol), DCC (107.3 mg, 0.52 mmol), DMAP (31.8 mg, 0.26 mmol), and DCM (2 mL) were added to a 25 mL flask, and the mixture was stirred at room temperature for 30 min. 56-2 (384.9 mg, 0.52 mmol) was then added dropwise, and the mixture was stirred at room temperature overnight. The extent of the reaction was monitored by TLC (5% MeOH / DCM). After completion of the reaction, the mixture was filtered, and the filtrate was washed with 5 mL of water and 5 mL of saturated brine, dried over anhydrous Na2SO4, and filtered. The organic phase was concentrated to give the crude product (approximately 1 g). The crude product was purified and separated by flash column (silica gel 60 g; mobile phases were 100 mL of n-heptane; 100 mL of 2% EtOAc + 98% n-heptane; 100 mL of 8% EtOAc + 92% n-heptane; and 500 mL of 20% EtOAc + 80% n-heptane) to give the pure product (450 mg, 78% yield). 1 H NMR (400 MHz, CDCl3): δ 7.67-7.65 (m, 4H), 7.44-7.26 (m, 6H), 5.13-5.12 (m, 1H), 4.89-4.83 (m, 1H), 4.35-4.31 (m, 1H), 4.09-4.03 (m, 3H), 3.72-3.69 (m, 2H), 2.65-2.55 (m, 3H), 2.34-2.22 (m, 10H), 1.65-1.19 (m, 72H), 1.04 (s, 9H), 0.89-0.85 (m, 12H).

[0205] Step 2: Synthesis of compound 56 56-3 (450 mg, 0.41 mmol) and THF (3 mL) were added to a 25 mL flask, and the mixture was cooled to 0–5 °C in an ice bath. TBAF (318.9 mg, 1.22 mmol) was added dropwise while maintaining the temperature at 0–5 °C, and the mixture was stirred at this temperature for 3 h. The extent of the reaction was monitored by TLC (5% MeOH / DCM). After completion of the reaction, the reaction was quenched with saturated aqueous ammonium chloride solution, and the mixture was extracted with EtOAc (5 mL × 2). The organic phases were combined, dried over anhydrous Na2SO4, filtered, and concentrated to give the crude product (approximately 530 mg). The crude product was purified and separated by flash column (50 g of silica gel packed with 100 mL of n-heptane; the mobile phases were 100 mL of 1% EtOAc+99% n-heptane; 100 mL of 2% EtOAc+98% n-heptane; 100 mL of 10% EtOAc+90% n-heptane; and 300 mL of 25% EtOAc+75% n-heptane), to give the pure product (240 mg, 68% yield). 1 H NMR (400 MHz, CDCl3): δ 5.23-5.18 (m, 1H), 4.88-4.82 (m, 1H), 4.35-4.31 (m, 1H), 4.12-4.03 (m, 1H), 3.58 (t, J=4Hz, 2H), 2.66-2.56 (m, 5H), 2.34-2.25 (m, 10H), 1.66-1.25 (m, 68H), 0.88-0.85 (m, 12H).

[0206] Synthesis of compound 57 [ka]

[0207] Step 1: Synthesis of compound 57-3 57-1 (500 mg, 1.26 mmol), DCC (259.4 mg, 1.26 mmol), DMAP (153.6 mg, 1.26 mmol), and DCM (5 mL) were added to a 25 mL flask, and the mixture was stirred at room temperature for 30 min. 57-2 (913.1 mg, 1.26 mmol) was then added dropwise, and the mixture was stirred at room temperature overnight. The extent of the reaction was monitored by TLC (5% MeOH / DCM). After completion of the reaction, the mixture was filtered, and the filtrate was washed with 5 mL of water and 5 mL of saturated brine, dried over anhydrous Na2SO4, and filtered. The organic phase was concentrated to give the crude product (approximately 1 g). The crude product was purified and separated by flash column (silica gel 60 g; mobile phases were 100 mL each of n-heptane; 100 mL of 2% EtOAc + 98% n-heptane; 100 mL of 8% EtOAc + 92% n-heptane; 100 mL of 20% EtOAc + 80% n-heptane; and 400 mL of 50% EtOAc + 50% n-heptane) to give the pure product (600 mg, 43% yield). 1 H NMR (400 MHz, CDCl3): δ 7.68-7.65 (m, 4H), 7.42-7.35 (m, 6H), 5.13 (m, 1H), 4.87-4.84 (m, 1H), 4.35-4.31 (m, 1H), 4.09-4.05 (m, 1H), 3.72-3.69 (m, 2H), 3.26-3.21 (m, 2H), 2.61-2.55 (m, 3H), 2.29-2.24 (m, 10H), 1.67-1.46 (m, 15H), 1.39-1.19 (m, 56H), 1.03 (s, 9H), 0.89-0.85 (m, 12H).

[0208] Step 2: Synthesis of compound 57 57-3 (550 mg, 0.5 mmol) and THF (3 mL) were added to a 25 mL flask, and the mixture was cooled to 0–5 °C in an ice bath. TBAF (390.2 mg, 1.5 mmol) was added dropwise while maintaining the temperature at 0–5 °C, and the mixture was stirred at this temperature for 3 h. The extent of the reaction was monitored by TLC (5% MeOH / DCM). After completion of the reaction, the reaction was quenched with saturated aqueous ammonium chloride solution, and the mixture was extracted with EtOAc (5 mL × 2). The organic phases were combined, dried over anhydrous Na2SO4, filtered, and concentrated to give the crude product (approximately 500 mg). The crude product was purified and separated by flash column (50 g of silica gel packed with 100 mL of n-heptane; the mobile phases were 100 mL of 1% EtOAc+99% n-heptane; 100 mL of 2% EtOAc+98% n-heptane; 100 mL of 10% EtOAc+90% n-heptane; 100 mL of 20% EtOAc+80% n-heptane; and 300 mL of 50% EtOAc+50% n-heptane), to give the pure product (200 mg, 46% yield). 1 H NMR (400 MHz, CDCl3): δ 5.60-5.56 (m, 1H), 5.22-5.18 (m, 1H), 4.88-4.81 (m, 1H), 4.35-4.30 (m, 1H), 3.57 (t, J=4Hz, 2H), 3.26-3.21 (m, 2H), 2.64-2.56 (m, 5H), 2.32-2.25 (m, 10H), 1.64-1.19 (m, 72H), 0.88-0.84 (m, 12H).

[0209] Synthesis of compound 58 [ka]

[0210] Step 1: Synthesis of compound 58-3 58-1 (300 mg, 0.75 mmol), DCC (155.3 mg, 0.75 mmol), DMAP (91.9 mg, 0.75 mmol), and DCM (3 mL) were added to a 25 mL flask, and the mixture was stirred at room temperature for 30 min. 58-2 (557 mg, 0.75 mmol) was then added dropwise, and the mixture was stirred at room temperature overnight. The extent of the reaction was monitored by TLC (5% MeOH / DCM). After completion of the reaction, the mixture was filtered, and the filtrate was washed with 5 mL of water and 5 mL of saturated brine, dried over anhydrous Na2SO4, and filtered. The organic phase was concentrated to give the crude product (approximately 700 mg). The crude product was purified and separated by flash column (50 g silica gel; mobile phases were 100 mL of n-heptane; 100 mL of 2% EtOAc + 98% n-heptane; 100 mL of 8% EtOAc + 92% n-heptane; and 500 mL of 20% EtOAc + 80% n-heptane) to give the pure product (500 mg, 59% yield). 1 H NMR (400 MHz, CDCl3): δ 7.69-7.66 (m, 4H), 7.45-7.37 (m, 6H), 4.16-4.12 (m, 1H), 4.07-3.97 (m, 3H), 3.88-3.71 (m, 4H), 2.75-2.57 (m, 3H), 2.47-2.46 (m, 2H), 2.36-2.28 (m, 7H), 1.65-1.55 (m, 7H), 1.46-1.20 (m, 34H), 1.05 (s, 9H), 0.90-0.85 (m, 12H).

[0211] Step 2: Synthesis of compound 58 58-3 (520 mg, 0.46 mmol) and THF (3 mL) were added to a 25 mL flask, and the mixture was cooled to 0–5 °C in an ice bath. TBAF (363.9 mg, 1.39 mmol) was added dropwise while maintaining the temperature at 0–5 °C, and the mixture was stirred at this temperature for 3 h. The extent of the reaction was monitored by TLC (5% MeOH / DCM). After completion of the reaction, the reaction was quenched with saturated aqueous ammonium chloride solution, and the mixture was extracted with EtOAc (5 mL × 2). The organic phases were combined, dried over anhydrous Na2SO4, filtered, and concentrated to give the crude product (approximately 500 mg). The crude product was purified and separated by flash column (50 g of silica gel packed with 100 mL of n-heptane; the mobile phases were 100 mL of 1% EtOAc + 99% n-heptane; 100 mL of 2% EtOAc + 98% n-heptane; 100 mL of 10% EtOAc + 90% n-heptane; and 300 mL of 20% EtOAc + 80% n-heptane), to give the pure product (230 mg, 56% yield). 1 H NMR (400 MHz, CDCl3): δ 5.23-5.18 (m, 1H), 4.88-4.82 (m, 1H), 4.34-4.31 (m, 1H), 4.12-4.03 (m, 3H), 3.57 (t, J=4Hz, 2H), 2.68-2.56 (m, 5H), 2.33-2.25 (m, 10H), 1.65-1.25 (m, 72H), 0.88-0.84 (m, 12H).

[0212] Synthesis of compound 59 [ka]

[0213] Step 1: Synthesis of compound 59-3 59-1 (204.9 mg, 0.65 mmol), DCC (134.5 mg, 0.65 mmol), DMAP (79.62 mg, 0.65 mmol), and DCM (3 mL) were added to a 25 mL flask, and the mixture was stirred at room temperature for 30 min. 59-2 (500 mg, 0.65 mmol) was then added dropwise, and the mixture was stirred at room temperature overnight. The extent of the reaction was monitored by TLC (5% MeOH / DCM). After completion of the reaction, the mixture was filtered, and the filtrate was washed with 5 mL of water and 5 mL of saturated brine, dried over anhydrous Na2SO4, and filtered. The organic phase was concentrated to give the crude product (approximately 1 g). The crude product was purified and separated by flash column (silica gel 60 g; mobile phases were 100 mL each of n-heptane; 100 mL of 1% EtOAc + 99% n-heptane; 100 mL of 2% EtOAc + 98% n-heptane; 100 mL of 8% EtOAc + 92% n-heptane; and 500 mL of 20% EtOAc + 80% n-heptane) to give the pure product (440 mg, 63% yield). 1 H NMR (400 MHz, CDCl3): δ 7.67-7.65 (m, 4H), 7.44-7.36 (m, 6H), 4.16-4.12 (m, 1H), 5.12 (m, 1H), 4.91-4.86 (m, 1H), 4.34-4.30 (m, 1H), 4.09-4.04 (m, 1H), 3.71 (m, 2H), 3.29-3.16 (m, 4H), 2.60-2.55 (m, 4H), 2.29-2.23 (m, 10H), 1.66-1.42 (m, 17H), 1.37-1.18 (m, 50H), 1.04 (s, 9H), 0.90-0.85 (m, 9H).

[0214] Step 2: Synthesis of compound 59 59-3 (440 mg, 0.41 mmol) and THF (3 mL) were added to a 25 mL flask, and the mixture was cooled to 0–5 °C in an ice bath. TBAF (324.5 mg, 1.24 mmol) was added dropwise while maintaining the temperature at 0–5 °C, and the mixture was stirred at this temperature for 3 h. The extent of the reaction was monitored by TLC (5% MeOH / DCM). After completion of the reaction, the reaction was quenched with saturated aqueous ammonium chloride solution, and the mixture was extracted with EtOAc (5 mL × 2). The organic phases were combined, dried over anhydrous Na2SO4, filtered, and concentrated to give the crude product (approximately 530 mg). The crude product was purified and separated by flash column (50 g of silica gel packed with 100 mL of n-heptane; mobile phases were 100 mL of 1% EtOAc+99% n-heptane; 100 mL of 2% EtOAc+98% n-heptane; 100 mL of 10% EtOAc+90% n-heptane; 100 mL of 30% EtOAc+70% n-heptane; and 400 mL of 50% EtOAc+50% n-heptane), to give the pure product (190 mg, 56% yield). 1 H NMR (400 MHz, CDCl3): δ 5.23-5.18 (m, 1H), 4.90-4.85 (m, 1H), 4.34-4.30 (m, 1H), 4.12-4.07 (m, 3H), 3.59 (t, J=4Hz, 2H), 3.28-3.16 (m, 4H), 2.65-2.59 (m, 5H), 2.34-2.23 (m, 10H), 1.65-1.17 (m, 62H), 0.88-0.84 (m, 9H).

[0215] Synthesis of compound 60 [ka]

[0216] Step 1: Synthesis of compound 60-2 4-(Methylamino)butan-1-ol (60-1, 2.0 g, 19.4 mmol) and DCM (10 mL) were added to a 100 mL flask, and the solution was cooled to 0-5 °C in an ice-salt bath. To the cooled solution, a solution of TBDPSCl (5.33 g, 19.4 mmol) in DCM (5 mL) was slowly added dropwise. After the addition, the mixture was stirred at room temperature for 2 h. The extent of the reaction was monitored by TLC (10% MeOH / DCM). After completion of the reaction, the reaction solution was concentrated to dryness to obtain the crude product (approximately 8 g). The crude product was purified and separated by flash column (70 g silica gel packed with 100 mL of n-heptane; mobile phases were 200 mL of n-heptane; 200 mL of 0.1% EtOAc + 99.9% n-heptane; 200 mL of 0.3% EtOAc + 99.7% n-heptane; 200 mL of 1% EtOAc + 99% n-heptane; 200 mL of 1% EtOAc + 99% n-heptane; 200 mL of 2% MeOH + 98% DCM; and 300 mL of 10% MeOH + 90% DCM) to give the pure product (5.5 g, 83% yield).

[0217] Step 2: Synthesis of compound 60-3 60-2 (5.0 g, 14.6 mmol), potassium carbonate (5.1 g, 36.6 mmol), and 3-chloro-1,2-propanediol (1.94 g, 17.6 mmol) were dissolved in isopropanol (15 mL) and added to a 100 mL flask. The mixture was heated to 80 °C and stirred overnight. The extent of the reaction was monitored by TLC (10% MeOH / DCM). After completion of the reaction, the mixture was filtered, and the organic phase was concentrated to give the crude product (approximately 5.3 g). The crude product was purified and separated by flash column (60 g silica gel; 100 mL of 0.1% MeOH+99.9% DCM; 100 mL of 0.2% MeOH+99.8% DCM; 100 mL of 0.3% MeOH+99.7% DCM; and 300 mL of 0.5% MeOH+99.5% DCM) to give the pure product (3.2 g, 53% yield).

[0218] Step 3: Synthesis of compound 60-5 60-4 (350 mg, 0.88 mmol), DCC (181.2 mg, 0.44 mmol), DMAP (182.5 mg, 0.44 mmol), and DCM (5 mL) were added to a 25 mL flask, and the mixture was stirred at room temperature for 30 min. 60-3 (486.2 mg, 1.25 mmol) was then added dropwise, and the mixture was stirred at room temperature overnight. The extent of the reaction was monitored by TLC (20% EtOAc / n-heptane). After completion of the reaction, the mixture was filtered, and the filtrate was washed with 5 mL of water and 5 mL of saturated brine, dried over anhydrous Na2SO4, and filtered. The organic phase was concentrated to give the crude product (approximately 1 g). The crude product was purified and separated by flash column (silica gel 30 g; mobile phases were 100 mL each of n-heptane; 100 mL of 1% EtOAc + 99% n-heptane; 100 mL of 2% EtOAc + 98% n-heptane; 100 mL of 8% EtOAc + 92% n-heptane; and 300 mL of 30% EtOAc + 70% n-heptane) to give the pure product (290 mg, 56% yield).

[0219] Step 4: Synthesis of compound 60-7 60-6 (145 mg, 0.36 mmol), DCC (75.2 mg, 0.36 mmol), DMAP (44.6 mg, 0.36 mmol), and DCM (5 mL) were added to a 25 mL flask, and the mixture was stirred at room temperature for 30 min. 60-5 (290 mg, 0.36 mmol) was then added dropwise, and the mixture was stirred at room temperature overnight. The extent of the reaction was monitored by TLC (20% EtOAc / n-heptane). After completion of the reaction, the mixture was filtered, and the filtrate was washed with 25 mL of water and 25 mL of saturated brine, dried over anhydrous Na2SO4, and filtered. The organic phase was concentrated to give the crude product (approximately 600 mg). The crude product was purified and separated by flash column (30 g silica gel; mobile phases were 100 mL each of n-heptane; 100 mL of 1% EtOAc + 99% n-heptane; 100 mL of 2% EtOAc + 98% n-heptane; 100 mL of 8% EtOAc + 92% n-heptane; and 100 mL of 30% EtOAc + 70% n-heptane) to give the pure product (60 mg, 14% yield).

[0220] Step 5: Synthesis of Compound 60 60-7 (550 mg, 0.47 mmol) and THF (3 mL) were added to a 25 mL flask, and the mixture was cooled to 0–5 °C in an ice bath. TBAF (366.6 mg, 1.4 mmol) was added dropwise while maintaining the temperature at 0–5 °C, and the mixture was stirred at this temperature for 3 h. The extent of the reaction was monitored by TLC (5% MeOH / DCM). After completion of the reaction, the reaction was quenched with saturated aqueous ammonium chloride solution, and the mixture was extracted with EtOAc (5 mL × 2). The organic phases were combined, dried over anhydrous Na2SO4, filtered, and concentrated to give the crude product (approximately 830 mg). The crude product was purified and separated by flash column (50 g of silica gel packed with 100 mL of n-heptane; the mobile phases were 100 mL of 1% EtOAc + 99% n-heptane; 100 mL of 2% EtOAc + 98% n-heptane; 100 mL of 10% EtOAc + 90% n-heptane; and 400 mL of 30% EtOAc + 70% n-heptane), to give the pure product (200 mg, 46% yield). 1 H NMR (400 MHz, CDCl3): δ 5.29-5.25 (m, 1H), 4.35-4.31 (m, 1H), 4.08-4.03 (m, 5H), 3.58-3.55 (m, 2H), 2.70-2.65 (m, 1H), 2.52-2.26 (m, 12H), 1.69-1.53 ​​(m, 16H), 1.44-1.25 (m, 52H), 0.89-0.85 (m, 12H).

[0221] Synthesis of compound 63 [ka]

[0222] Step 1: Synthesis of compound 63-3 63-1 (200 mg, 0.52 mmol), DCC (107.3 mg, 0.52 mmol), DMAP (31.8 mg, 0.26 mmol), and DCM (5 mL) were added to a 25 mL flask, and the mixture was stirred at room temperature for 30 min. 63-2 (107.8 mg, 0.26 mmol) was then added, and the mixture was stirred at room temperature overnight. The extent of the reaction was monitored by TLC (5% MeOH / DCM). After completion of the reaction, the mixture was filtered, and the filtrate was washed with 25 mL of water and 25 mL of saturated brine, dried over anhydrous Na2SO4, and filtered. The organic phase was concentrated to give the crude product (approximately 300 mg). The crude product was purified and separated by flash column (50 g silica gel; mobile phases were 100 mL of n-heptane; 100 mL of 2% EtOAc + 98% n-heptane; 100 mL of 8% EtOAc + 92% n-heptane; and 200 mL of 20% EtOAc + 80% n-heptane) to give the pure product (130 mg, 43% yield).

[0223] Step 2: Synthesis of compound 63 63-3 (130 mg, 0.11 mmol) and THF (3 mL) were added to a 25 mL flask, and the mixture was cooled to 0–5 °C in an ice bath. TBAF (88.8 mg, 0.33 mmol) was added dropwise while maintaining the temperature at 0–5 °C, and the mixture was stirred at this temperature for 3 h. The extent of the reaction was monitored by TLC (5% MeOH / DCM). After completion of the reaction, the reaction was quenched with saturated aqueous ammonium chloride solution, and the mixture was extracted with EtOAc (5 mL × 2). The organic phases were combined, dried over anhydrous Na2SO4, filtered, and concentrated to give the crude product (approximately 500 mg). The crude product was purified and separated by flash column (50 g of silica gel packed with 100 mL of n-heptane; the mobile phases were 100 mL of 1% EtOAc + 99% n-heptane; 100 mL of 2% EtOAc + 98% n-heptane; 100 mL of 10% EtOAc + 90% n-heptane; and 200 mL of 20% EtOAc + 80% n-heptane), respectively, to give the pure product (60 mg, 60% yield). 1H NMR (400 MHz, CDCl3): δ 5.29-5.25 (m, 1H), 4.88-4.82 (m, 2H), 4.34-4.30 (m, 1H), 4.08-4.03 (m, 1H), 3.58-3.55 (m, 2H), 2.71-2.66 (m, 1H), 2.54-2.21 (m, 14H), 1.65-1.47 (m, 20H), 1.36-1.25 (m, 52H), 0.89-0.85 (m, 12H).

[0224] Synthesis of compound 64 [ka]

[0225] Step 1: Synthesis of compound 64-3 64-1 (400 mg, 1.04 mmol), DCC (214.6 mg, 0.52 mmol), DMAP (63.5 mg, 0.52 mmol), and DCM (5 mL) were added to a 25 mL flask, and the mixture was stirred at room temperature for 30 min. 64-2 (215.6 mg, 0.52 mmol) was then added, and the mixture was stirred at room temperature overnight. The extent of the reaction was monitored by TLC (5% MeOH / DCM). After completion of the reaction, the mixture was filtered, and the filtrate was washed with 25 mL of water and 25 mL of saturated brine, dried over anhydrous Na2SO4, and filtered. The organic phase was concentrated to give the crude product (approximately 500 mg). The crude product was purified and separated by flash column (50 g silica gel; mobile phases were 100 mL of n-heptane; 100 mL of 2% EtOAc + 98% n-heptane; 100 mL of 8% EtOAc + 92% n-heptane; and 200 mL of 20% EtOAc + 80% n-heptane), to give the pure product (220 mg, 37% yield).

[0226] Step 2: Synthesis of compound 64 64-3 (220 mg, 0.19 mmol) and THF (3 mL) were added to a 25 mL flask, and the mixture was cooled to 0–5 °C in an ice bath. TBAF (150.2 mg, 0.57 mmol) was added dropwise while maintaining the temperature at 0–5 °C, and the mixture was stirred at this temperature for 3 h. The extent of the reaction was monitored by TLC (5% MeOH / DCM). After completion of the reaction, the reaction was quenched with saturated aqueous ammonium chloride solution, and the mixture was extracted with EtOAc (5 mL × 2). The organic phases were combined, dried over anhydrous Na2SO4, filtered, and concentrated to give the crude product (approximately 200 mg). The crude product was purified and separated by flash column (50 g of silica gel packed with 100 mL of n-heptane; the mobile phases were 100 mL of 1% EtOAc + 99% n-heptane; 100 mL of 2% EtOAc + 98% n-heptane; 100 mL of 10% EtOAc + 90% n-heptane; and 200 mL of 20% EtOAc + 80% n-heptane), to give the pure product (80 mg, 46% yield). 1 H NMR (400 MHz, CDCl3): δ 5.29-5.25 (m, 1H), 4.35-4.31 (m, 1H), 4.08-4.03 (m, 5H), 3.58-3.55 (m, 2H), 2.72-2.66 (m, 1H), 2.54-2.26 (m, 12H), 1.65-1.25 (m, 72H), 0.88-0.85 (m, 12H).

[0227] Synthesis of compound 65 [ka]

[0228] Step 1: Synthesis of compound 65-2 Pyrrolidine (65-1, 5.0 g, 70.3 mmol), potassium carbonate (24.3 g, 175.8 mmol), and 3-chloro-1,2-propanediol (7.77 g, 70.3 mmol) were dissolved in isopropanol (50 mL) and added to a 250 mL flask. The mixture was heated to 80 °C and stirred overnight. The extent of the reaction was monitored by TLC (10% MeOH / DCM). After completion of the reaction, the mixture was filtered, and the organic phase was concentrated to give the crude product (approximately 1.5 g, 15% yield).

[0229] Step 2: Synthesis of compound 65 65-3 (1.0 g, 2.51 mmol), DCC (517.6 mg, 2.51 mmol), DMAP (153.2 mg, 1.25 mmol), and DCM (3 mL) were added to a 25 mL flask, and the mixture was stirred at room temperature for 30 min. 65-2 (182.1 mg, 1.25 mmol) was then added dropwise, and the mixture was stirred at room temperature overnight. The extent of the reaction was monitored by TLC (20% EtOAc / n-heptane). After completion of the reaction, the mixture was filtered, and the filtrate was washed with 5 mL of water and 5 mL of saturated brine, dried over anhydrous Na2SO4, and filtered. The organic phase was concentrated to give the crude product (approximately 600 mg). The crude product was purified and separated by flash column (silica gel 30 g; mobile phases were 100 mL of n-heptane; 100 mL of 1% EtOAc + 99% n-heptane; 100 mL of 2% EtOAc + 98% n-heptane; 100 mL of 8% EtOAc + 92% n-heptane; and 400 mL of 30% EtOAc + 70% n-heptane) to give the pure product (380 mg, 33% yield). 1 H NMR (400 MHz, CDCl3): δ 5.24-5.19 (m, 1H), 4.40-4.37 (m, 1H), 4.12-4.04 (m, 5H), 2.65-2.55 (m, 6H), 2.34-2.28 (m, 6H), 1.77-1.53 (m, 17H), 1.46-1.19 (m, 51H), 0.89-0.85 (m, 12H).

[0230] Synthesis of compound 66 [ka]

[0231] Step 1: Synthesis of compound 66-2 Morpholine (66-1, 5.0 g, 57.4 mmol), potassium carbonate (19.8 g, 143.5 mmol), and 3-chloro-1,2-propanediol (6.34 g, 57.4 mmol) were dissolved in isopropanol (50 mL) and added to a 250 mL flask. The mixture was heated to 80 °C and stirred overnight. The extent of the reaction was monitored by TLC (10% MeOH / DCM). After completion of the reaction, the mixture was filtered, and the organic phase was concentrated to give the crude product (approximately 2.2 g, 24% yield).

[0232] Step 2: Synthesis of compound 66 66-3 (500 mg, 1.35 mmol), DCC (278.4 mg, 1.35 mmol), DMAP (82.4 mg, 0.67 mmol), and DCM (2 mL) were added to a 25 mL flask, and the mixture was stirred at room temperature for 30 min. 66-2 (108.75 mg, 0.67 mmol) was then added dropwise, and the mixture was stirred at room temperature overnight. The extent of the reaction was monitored by TLC (20% EtOAc / n-heptane). After completion of the reaction, the mixture was filtered, and the filtrate was washed with 5 mL of water and 5 mL of saturated brine, dried over anhydrous Na2SO4, and filtered. The organic phase was concentrated to give the crude product (approximately 600 mg). The crude product was purified and separated by flash column (30 g silica gel; mobile phases were 100 mL of n-heptane; 100 mL of 1% EtOAc + 99% n-heptane; 100 mL of 2% EtOAc + 98% n-heptane; 100 mL of 8% EtOAc + 92% n-heptane; and 200 mL of 30% EtOAc + 70% n-heptane) to give the pure product (300 mg, 51% yield). 1H NMR (400 MHz, CDCl3): δ 5.23 (m, 1H), 4.39-4.35 (m, 1H), 4.12-4.04 (m, 5H), 3.66 (m, 4H), 2.49 (m, 6H), 2.34-2.26 (m, 6H), 1.70-1.53 ​​(m, 14H), 1.46-1.19 (m, 54H), 0.89-0.85 (m, 12H).

[0233] Synthesis of compound 67 [ka]

[0234] Step 1: Synthesis of compound 67-2 Piperidine (3.0 g, 35.2 mmol), potassium carbonate (12.2 g, 88.1 mmol), and 3-chloro-1,2-propanediol (67-1, 3.89 g, 35.2 mmol) were dissolved in isopropanol (20 mL) and added to a 100 mL flask. The mixture was heated to 80 °C and stirred overnight. The extent of the reaction was monitored by TLC (10% MeOH / DCM). After completion of the reaction, the mixture was filtered, and the organic phase was concentrated to give the crude product (approximately 3.2 g, 57% yield).

[0235] Step 2: Synthesis of compound 67 67-3 (512 mg, 1.38 mmol), DCC (285.1 mg, 1.38 mmol), DMAP (84.4 mg, 0.69 mmol), and DCM (3 mL) were added to a 25 mL flask, and the mixture was stirred at room temperature for 30 min. 67-2 (110 mg, 0.69 mmol) was then added dropwise, and the mixture was stirred at room temperature overnight. The extent of the reaction was monitored by TLC (20% EtOAc / n-heptane). After completion of the reaction, the mixture was filtered, and the filtrate was washed with 5 mL of water and 5 mL of saturated brine, dried over anhydrous Na2SO4, and filtered. The organic phase was concentrated to give the crude product (approximately 700 mg). The crude product was purified and separated by flash column (silica gel 20 g; mobile phases were 100 mL of n-heptane; 100 mL of 1% EtOAc + 99% n-heptane; 100 mL of 2% EtOAc + 98% n-heptane; 100 mL of 8% EtOAc + 92% n-heptane; and 200 mL of 20% EtOAc + 80% n-heptane) to give the pure product (250 mg, 42% yield). 1 H NMR (400 MHz, CDCl3): δ 5.22 (m, 1H), 4.37-4.33 (m, 1H), 4.12-4.04 (m, 5H), 2.44-2.26 (m, 12H), 1.70-1.19 (m, 76H), 0.89-0.85 (m, 12H).

[0236] Synthesis of compound 68 [ka]

[0237] Step 1: Synthesis of compound 68-2 Diethanolamine (68-1, 2.0 g, 19 mmol) and DCM (20 mL) were added to a 100 mL flask, and the solution was cooled to 0-5 °C in an ice-salt bath. To the cooled solution, a solution of TBDPSCl (10.5 g, 38 mmol) in DCM (5 mL) was slowly added dropwise. After the addition, the mixture was stirred at room temperature for 2 h. The extent of the reaction was monitored by TLC (20% MeOH / DCM). After completion of the reaction, the reaction solution was concentrated to dryness to obtain the crude product (approximately 20 g). The crude product was purified and separated by flash column (200 g silica gel packed with 100 mL of n-heptane; mobile phases were 200 mL of n-heptane; 200 mL of 0.1% EtOAc + 99.9% n-heptane; 200 mL of 0.3% EtOAc + 99.7% n-heptane; 200 mL of 1% EtOAc + 99% n-heptane; 200 mL of 1% EtOAc + 99% n-heptane; 200 mL of 2% MeOH + 98% DCM; and 300 mL of 10% MeOH + 90% DCM) to give the pure product (6 g, 54% yield).

[0238] Step 2: Synthesis of compound 68-3 68-2 (3.0 g, 5.2 mmol), potassium carbonate (1.8 g, 12.9 mmol), and 3-chloro-1,2-propanediol (0.57 g, 5.2 mmol) were dissolved in isopropanol (10 mL) and added to a 50 mL flask. The mixture was heated to 80 °C and stirred overnight. The reaction progress was monitored by TLC (10% MeOH / DCM). After completion of the reaction, the mixture was filtered, and the organic phase was concentrated to give the crude product (approximately 4 g). The crude product was purified and separated by flash column (50 g silica gel; 100 mL of 0.1% MeOH+99.9% DCM; 100 mL of 0.2% MeOH+99.8% DCM; 100 mL of 0.3% MeOH+99.7% DCM; and 300 mL of 0.5% MeOH+99.5% DCM) to give the pure product (2.5 g, 74% yield).

[0239] Step 3: Synthesis of compound 68-5 68-4 (564.9 mg, 1.5 mmol), DCC (314.5 mg, 1.5 mmol), DMAP (93.1 mg, 0.76 mmol), and DCM (5 mL) were added to a 25 mL flask, and the mixture was stirred at room temperature for 30 min. 68-3 (500 mg, 0.76 mmol) was then added dropwise, and the mixture was stirred at room temperature overnight. The extent of the reaction was monitored by TLC (20% EtOAc / n-heptane). After completion of the reaction, the mixture was filtered, and the filtrate was washed with 5 mL of water and 5 mL of saturated brine, dried over anhydrous Na2SO4, and filtered. The organic phase was concentrated to give the crude product (approximately 1.2 g). The crude product was purified and separated by flash column (12 g silica gel; mobile phases were 100 mL of n-heptane; 100 mL of 1% EtOAc + 99% n-heptane; 100 mL of 2% EtOAc + 98% n-heptane; and 100 mL of 8% EtOAc + 92% n-heptane), to give the pure product (680 mg, 67% yield).

[0240] Step 4: Synthesis of compound 68 68-5 (680 mg, 0.52 mmol) and THF (3 mL) were added to a 25 mL flask, and the mixture was cooled to 0–5 °C in an ice bath. TBAF (404.4 mg, 1.6 mmol) was added dropwise while maintaining the temperature at 0–5 °C, and the mixture was stirred at this temperature for 3 h. The extent of the reaction was monitored by TLC (5% MeOH / DCM). After completion of the reaction, the reaction was quenched with saturated aqueous ammonium chloride solution, and the mixture was extracted with EtOAc (5 mL × 2). The organic phases were combined, dried over anhydrous Na2SO4, filtered, and concentrated to give the crude product (approximately 500 mg). The crude product was purified and separated by flash column (12 g silica gel packed with 100 mL of n-heptane; mobile phases were 100 mL of 1% EtOAc + 99% n-heptane; 100 mL of 2% EtOAc + 98% n-heptane; 100 mL of 10% EtOAc + 90% n-heptane; 100 mL of 30% EtOAc + 70% n-heptane; and 1000 mL of 50% EtOAc + 50% n-heptane) to give the pure product (200 mg, 48% yield). 1H NMR (400 MHz, CDCl3): δ 5.29-5.25 (m, 1H), 4.35-4.31 (m, 1H), 4.08-4.03 (m, 5H), 3.58-3.55 (m, 2H), 2.70-2.56 (m, 1H), 2.52-2.26 (m, 12H), 1.69-1.53 ​​(m, 16H), 1.44-1.22 (m, 50H), 0.89-0.85 (m, 12H).

[0241] Synthesis of compound 70 [ka]

[0242] Step 1: Synthesis of compound 70-2 2-[2-(methylamino)ethoxy]ethan-1-ol (70-1, 500 mg, 4.2 mmol) and DCM (3 mL) were added to a 25 mL flask, and the solution was cooled to 0-5 °C in an ice-salt bath. To the cooled solution, a solution of TBDPSCl (1.15 g, 4.2 mmol) in DCM (2 mL) was slowly added dropwise. After the addition, the mixture was stirred at room temperature for 2 h. The extent of the reaction was monitored by TLC (20% MeOH / DCM). After completion of the reaction, the reaction solution was concentrated to dryness to obtain the crude product (approximately 2 g). The crude product was purified and separated by flash column (14 g silica gel packed with 100 mL of n-heptane; mobile phases were 100 mL of n-heptane; 100 mL of 0.1% EtOAc + 99.9% n-heptane; 100 mL of 0.3% EtOAc + 99.7% n-heptane; 100 mL of 1% EtOAc + 99% n-heptane; 100 mL of 1% EtOAc + 99% n-heptane; 100 mL of 2% MeOH + 98% DCM; and 300 mL of 10% MeOH + 90% DCM) to give the pure product (800 mg, 53% yield).

[0243] Step 2: Synthesis of compound 70-3 70-2 (690 mg, 1.9 mmol), potassium carbonate (666.7 mg, 4.8 mmol), and 3-chloro-1,2-propanediol (213.3 mg, 1.9 mmol) were dissolved in isopropanol (5 mL) and added to a 25 mL flask. The mixture was heated to 80 °C and stirred overnight. The reaction progress was monitored by TLC (10% MeOH / DCM). After completion of the reaction, the mixture was filtered, and the organic phase was concentrated to give the crude product (approximately 1 g). The crude product was purified and separated by flash column (12 g silica gel; 100 mL of 0.1% MeOH+99.9% DCM; 100 mL of 0.2% MeOH+99.8% DCM; 100 mL of 0.3% MeOH+99.7% DCM; and 300 mL of 0.5% MeOH+99.5% DCM) to give the pure product (500 mg, 60% yield).

[0244] Step 3: Synthesis of compound 70-5 70-4 (858.5 mg, 2.4 mmol), DCC (478 mg, 2.4 mmol), DMAP (141.5 mg, 1.2 mmol), and DCM (5 mL) were added to a 25 mL flask, and the mixture was stirred at room temperature for 30 min. Then, 70-3 (500 mg, 1.2 mmol) was added dropwise, and the mixture was stirred at room temperature overnight. The extent of the reaction was monitored by TLC (20% EtOAc / n-heptane). After completion of the reaction, the mixture was filtered, and the filtrate was washed with 5 mL of water and 5 mL of saturated brine, dried over anhydrous Na2SO4, and filtered. The organic phase was concentrated to give the crude product (approximately 1.5 g). The crude product was purified and separated by flash column (12 g silica gel; mobile phases were 100 mL of n-heptane; 100 mL of 1% EtOAc + 99% n-heptane; 100 mL of 2% EtOAc + 98% n-heptane; and 100 mL of 8% EtOAc + 92% n-heptane) to give the pure product (650 mg, 49% yield).

[0245] Step 4: Synthesis of compound 70 70-5 (650 mg, 0.57 mmol) and THF (3 mL) were added to a 25 mL flask, and the mixture was cooled to 0–5 °C in an ice bath. TBAF (448.5 mg, 1.7 mmol) was added dropwise while maintaining the temperature at 0–5 °C, and the mixture was stirred at this temperature for 3 h. The extent of the reaction was monitored by TLC (5% MeOH / DCM). After completion of the reaction, the reaction was quenched with saturated aqueous ammonium chloride solution, and the mixture was extracted with EtOAc (5 mL × 2). The organic phases were combined, dried over anhydrous Na2SO4, filtered, and concentrated to give the crude product (approximately 700 mg). The crude product was purified and separated by flash column (12 g silica gel packed with 100 mL of n-heptane; mobile phases were 100 mL of 1% EtOAc + 99% n-heptane; 100 mL of 2% EtOAc + 98% n-heptane; 100 mL of 10% EtOAc + 90% n-heptane; and 100 mL of 30% EtOAc + 70% n-heptane), to give the pure product (250 mg, 49% yield). 1 H NMR (400 MHz, CDCl3): δ 5.25-5.20 (m, 1H), 4.40-4.36 (m, 1H), 4.12-4.04 (m, 5H), 3.70-3.68 (m, 2H), 3.60-3.55 (m, 4H), 2.68-2.60 (m, 4H), 2.35-2.26 (m, 9H), 1.69-1.53 ​​(m, 13H), 1.46-1.34 (m, 9H), 1.30-1.19 (m,44H), 0.88-0.85 (m, 12H).

[0246] Synthesis of compound 76 [ka] Step 1: Synthesis of compound 76-2 Suberic acid (3.6 g, 20.6 mmol), DCC (4.3 g, 20.6 mmol), DMAP (503.9 mg, 4.1 mmol), and DCM (10 mL) were added to a 25 mL flask, and the mixture was stirred at room temperature for 30 min. Then, 2-hexadecanol (76-1, 1 g, 4.1 mmol) was added dropwise, and the mixture was stirred at room temperature overnight. The extent of the reaction was monitored by TLC (30% EtOAc / n-heptane). After completion of the reaction, the mixture was filtered, and the filtrate was washed with 5 mL of water and 5 mL of saturated brine, dried over anhydrous Na2SO4, and filtered. The organic phase was concentrated to give the crude product (approximately 2.5 g). The crude product was purified and separated by flash column (25 g silica gel; mobile phases were 100 mL of n-heptane; 100 mL of 1% EtOAc+99% n-heptane; 100 mL of 2% EtOAc+98% n-heptane; 100 mL of 8% EtOAc+92% n-heptane; and 100 mL of 20% EtOAc+80% n-heptane), to give the pure product (800 mg, 49% yield).

[0247] Step 2: Synthesis of compound 76-4 76-2 (300 mg, 0.56 mmol), DCC (116.5 mg, 0.56 mmol), DMAP (69 mg, 0.56 mmol), and DCM (2 mL) were added to a 25 mL flask, and the mixture was stirred at room temperature for 30 min. 76-3 (218.8 mg, 0.56 mmol) was then added dropwise, and the mixture was stirred at room temperature overnight. The extent of the reaction was monitored by TLC (30% EtOAc / n-heptane). After completion of the reaction, the mixture was filtered, and the filtrate was washed with 5 mL of water and 5 mL of saturated brine, dried over anhydrous Na2SO4, and filtered. The organic phase was concentrated to give the crude product (approximately 500 mg). The crude product was purified and separated by flash column (silica gel 12 g; mobile phases were 100 mL of n-heptane; 100 mL of 1% EtOAc + 99% n-heptane; 100 mL of 2% EtOAc + 98% n-heptane; and 100 mL of 8% EtOAc + 92% n-heptane) to give the pure product (200 mg, 46% yield).

[0248] Step 3: Synthesis of compound 76-6 76-5 (138.4 mg, 0.26 mmol), DCC (53.7 mg, 0.26 mmol), DMAP (31.8 mg, 0.26 mmol), and DCM (3 mL) were added to a 25 mL flask, and the mixture was stirred at room temperature for 30 min. 76-4 (500 mg, 1.2 mmol) was then added dropwise, and the mixture was stirred at room temperature overnight. The extent of the reaction was monitored by TLC (20% EtOAc / n-heptane). After completion of the reaction, the mixture was filtered, and the filtrate was washed with 5 mL of water and 5 mL of saturated brine, dried over anhydrous Na2SO4, and filtered. The organic phase was concentrated to give the crude product (approximately 350 mg). The crude product was purified and separated by flash column (12 g silica gel; mobile phases were 100 mL of n-heptane; 100 mL of 1% EtOAc + 99% n-heptane; 100 mL of 2% EtOAc + 98% n-heptane; and 100 mL of 8% EtOAc + 92% n-heptane) to give the pure product (180 mg, 60% yield). 1 H NMR (400 MHz, CDCl3): δ 7.68-7.64 (m, 4H), 7.44-7.36 (m, 6H), 5.14 (m, 1H), 4.93-4.85 (m, 2H), 4.35-4.31 (m, 1H), 4.09-4.05 (m, 1H), 3.72 (m, 2H), 2.62-2.58 (m, 4H), 2.31-2.23 (m, 11H), 1.63-1.52(m, 11H), 1.49-1.41 (m, 4H), 1,35-1.25 (m, 62H), 1.21-1.18 (m, 6H), 1.04 (s, 9H), 0.89-0.86 (m, 6H).

[0249] Step 4: Synthesis of Compound 76 76-6 (180 mg, 0.16 mmol) and THF (3 mL) were added to a 25 mL flask, and the mixture was cooled to 0–5 °C in an ice bath. TBAF (122.9 mg, 0.47 mmol) was added dropwise while maintaining the temperature at 0–5 °C, and the mixture was stirred at this temperature for 3 h. The extent of the reaction was monitored by TLC (5% MeOH / DCM). After completion of the reaction, the reaction was quenched with saturated aqueous ammonium chloride solution, and the mixture was extracted with EtOAc (5 mL × 2). The organic phases were combined, dried over anhydrous Na2SO4, filtered, and concentrated to give the crude product (approximately 200 mg). The crude product was purified and separated by flash column (4 g of silica gel packed with 100 mL of n-heptane; the mobile phases were 100 mL of 1% EtOAc + 99% n-heptane; 100 mL of 2% EtOAc + 98% n-heptane; 100 mL of 10% EtOAc + 90% n-heptane; and 400 mL of 30% EtOAc + 70% n-heptane), to give the pure product (80 mg, 56% yield). 1 H NMR (400 MHz, CDCl3): δ 5.24-5.19 (m, 1H), 4.92-4.84 (m, 2H), 4.35-4.31 (m, 1H), 4.12-4.08 (m, 1H), 3.60-3.57 (m, 2H), 2.65-2.58 (m, 4H), 2.34-2.23 (m, 11H), 1.65-1.52(m, 11H), 1.48-1.18 (m, 71H), 0.89-0.86 (m, 6H).

[0250] Synthesis of compound 78 [ka]

[0251] Step 1: Synthesis of compound 78 78-2 (529.8 mg, 1.38 mmol), DCC (284.2 mg, 1.38 mmol), DMAP (84.1 mg, 0.69 mmol), and DCM (3 mL) were added to a 25 mL flask, and the mixture was stirred at room temperature for 30 min. 78-1 (100 mg, 0.69 mmol) was then added dropwise, and the mixture was stirred at room temperature overnight. The extent of the reaction was monitored by TLC (20% EtOAc / n-heptane). After completion of the reaction, the mixture was filtered, and the filtrate was washed with 5 mL of water and 5 mL of saturated brine, dried over anhydrous Na2SO4, and filtered. The organic phase was concentrated to give the crude product (approximately 800 mg). The crude product was purified and separated by flash column (10 g silica gel; mobile phases were 100 mL of n-heptane; 100 mL of 1% EtOAc + 99% n-heptane; 100 mL of 2% EtOAc + 98% n-heptane; 100 mL of 8% EtOAc + 92% n-heptane; and 200 mL of 20% EtOAc + 80% n-heptane) to give the pure product (200 mg, 33% yield). 1 H NMR (400 MHz, CDCl3): δ 5.23-5.18 (m, 1H), 4.89-4.83 (m, 2H), 4.39-4.36 (m, 1H), 4.12-4.07 (m, 1H), 2.66-2.55 (m, 6H), 2.33-2.25 (m, 8H), 1.77-1.74 (m, 2H), 1.65-1.47 (m, 17H), 1.37-1.25 (m, 55H), 0.89-0.85 (m, 12H).

[0252] Synthesis of compound 79 [ka]

[0253] Step 1: Synthesis of compound 79-3 79-2 (793.8 mg, 2.1 mmol), DCC (425.9 mg, 2.1 mmol), DMAP (126.1 mg, 1.05 mmol), and DCM (5 mL) were added to a 25 mL flask, and the mixture was stirred at room temperature for 30 min. 79-1 (400 mg, 1.05 mmol) was then added, and the mixture was stirred at room temperature overnight. The extent of the reaction was monitored by TLC (30% MeOH / DCM). After completion of the reaction, the mixture was filtered, and the filtrate was washed with 5 mL of water and 5 mL of saturated brine, dried over anhydrous Na2SO4, and filtered. The organic phase was concentrated to give the crude product (approximately 1.2 g). The crude product was purified and separated by flash column (silica gel 12 g; mobile phases were 100 mL of n-heptane; 100 mL of 2% EtOAc + 98% n-heptane; 100 mL of 8% EtOAc + 92% n-heptane; and 200 mL of 20% EtOAc + 80% n-heptane), to give the pure product (880 mg, 76% yield). 1 H NMR (400 MHz, CDCl3): δ 7.67-7.65 (m, 4H), 7.42-7.36 (m, 6H), 5.13 (m, 1H), 4.89-4.83 (m, 2H), 4.35-4.31 (m, 1H), 4.09-4.05 (m, 1H), 3.71 (m, 2H), 2.61-2.56 (m, 4H), 2.28-2.24 (m, 12H), 1.63-1.48 (m, 20H), 1.34-1.19 (m, 56H), 1.04 (s, 9H), 0.89-0.86 (m, 12H).

[0254] Step 2: Synthesis of compound 79 79-3 (880 mg, 0.79 mmol) and THF (3 mL) were added to a 25 mL flask, and the mixture was cooled to 0–5 °C in an ice bath. TBAF (623.7 mg, 2.4 mmol) was added dropwise while maintaining the temperature at 0–5 °C, and the mixture was stirred at this temperature for 4 h. The extent of the reaction was monitored by TLC (5% MeOH / DCM). After completion of the reaction, the reaction was quenched with saturated aqueous ammonium chloride solution, and the mixture was extracted with EtOAc (5 mL × 2). The organic phases were combined, dried over anhydrous Na2SO4, filtered, and concentrated to give the crude product (approximately 800 mg). The crude product was purified and separated by flash column (12 g silica gel packed with 100 mL of n-heptane; mobile phases were 100 mL of 1% EtOAc + 99% n-heptane; 100 mL of 2% EtOAc + 98% n-heptane; 100 mL of 10% EtOAc + 90% n-heptane; and 300 mL of 30% EtOAc + 70% n-heptane), to give the pure product (500 mg, 71% yield). 1 H NMR (400 MHz, CDCl3): δ 5.24-5.18 (m, 1H), 4.88-4.82 (m, 2H), 4.35-4.31 (m, 1H), 4.12-4.08 (m, 1H), 3.59-3.57 (m, 2H), 2.65-2.57 (m, 4H), 2.34-2.25 (m, 11H), 1.65-1.58 (m, 8H), 1.52.1.47 (m, 8H), 1.37-1.25 (m, 51H), 0.89-0.85 (m, 12H).

[0255] Synthesis of Compound 80 [ka]

[0256] Step 1: Synthesis of compound 80-3 80-2 (793.8 mg, 2.1 mmol), DCC (425.9 mg, 2.1 mmol), DMAP (126.1 mg, 1.05 mmol), and DCM (5 mL) were added to a 25 mL flask, and the mixture was stirred at room temperature for 30 min. 80-1 (400 mg, 1.05 mmol) was then added, and the mixture was stirred at room temperature overnight. The extent of the reaction was monitored by TLC (30% MeOH / DCM). After completion of the reaction, the mixture was filtered, and the filtrate was washed with 5 mL of water and 5 mL of saturated brine, dried over anhydrous Na2SO4, and filtered. The organic phase was concentrated to give the crude product (approximately 1.3 g). The crude product was purified and separated by flash column (25 g silica gel; mobile phases were 100 mL of n-heptane; 100 mL of 2% EtOAc + 98% n-heptane; 100 mL of 8% EtOAc + 92% n-heptane; and 200 mL of 20% EtOAc + 80% n-heptane), to give the pure product (770 mg, 61% yield).

[0257] Step 2: Synthesis of Compound 80 80-3 (770 mg, 0.7 mmol) and THF (3 mL) were added to a 25 mL flask, and the mixture was cooled to 0–5 °C in an ice bath. TBAF (545.7 mg, 2.1 mmol) was added dropwise while maintaining the temperature at 0–5 °C, and the mixture was stirred at this temperature for 4 h. The extent of the reaction was monitored by TLC (5% MeOH / DCM). After completion of the reaction, the reaction was quenched with saturated aqueous ammonium chloride solution, and the mixture was extracted with EtOAc (5 mL × 2). The organic phases were combined, dried over anhydrous Na2SO4, filtered, and concentrated to give the crude product (approximately 700 mg). The crude product was purified and separated by flash column (12 g silica gel packed with 100 mL of n-heptane; mobile phases were 100 mL of 1% EtOAc + 99% n-heptane; 100 mL of 2% EtOAc + 98% n-heptane; 100 mL of 10% EtOAc + 90% n-heptane; 100 mL of 30% EtOAc + 70% n-heptane; and 300 mL of 50% EtOAc + 50% n-heptane) to give the pure product (300 mg, 49% yield). 1H NMR (400 MHz, CDCl3): δ 5.63-5.59 (m, 1H), 5.38-5.29 (m, 4H), 4.59-4.49 (m, 2H), 4.16-4.06 (m, 1H), 3.82-3.75 (m, 1H), 3.53 (s, 9H), 2.49-2.41 (m, 2H), 2.05-1.86 (m, 11H), 1.68-1.60 (m, 2H), 1.43-1.12 (m, 52H), 0.89-0.85 (m, 6H).

[0258] Synthesis of compound 81 [ka]

[0259] Step 1: Synthesis of compound 81-2 4-Hydroxypiperidine (81-1, 2.0 g, 19.8 mmol) and DCM (20 mL) were added to a 100 mL flask, and the solution was cooled to 0-5 °C in an ice-salt bath. To the cooled solution, a solution of TBDPSCl (5.4 g, 19.8 mmol) in DCM (10 mL) was slowly added dropwise. After the addition, the mixture was stirred at room temperature for 2 h. The extent of the reaction was monitored by TLC (20% MeOH / DCM). After completion of the reaction, the reaction solution was concentrated to dryness to obtain the crude product (approximately 7 g). The crude product was purified and separated by flash column (150 g of silica gel packed with 100 mL of n-heptane; the mobile phases were 100 mL of n-heptane; 200 mL of 1% EtOAc + 99% n-heptane; 200 mL of 2% EtOAc + 98% n-heptane; and 400 mL of 8% EtOAc + 92% n-heptane) to give the pure product (4.2 g, 63% yield).

[0260] Step 2: Synthesis of compound 81-3 81-2 (1.5 g, 4.4 mmol), potassium carbonate (1.2 g, 8.8 mmol), and 3-chloro-1,2-propanediol (0.49 g, 4.4 mmol) were dissolved in isopropanol (5 mL) and added to a 25 mL flask. The mixture was heated to 80 °C and stirred overnight. The reaction progress was monitored by TLC (10% MeOH / DCM). After completion of the reaction, the mixture was filtered, and the organic phase was concentrated to give the crude product (approximately 2 g). The crude product was purified and separated by flash column (20 g silica gel; 100 mL of 0.1% MeOH+99.9% DCM; 100 mL of 0.2% MeOH+99.8% DCM; 100 mL of 0.3% MeOH+99.7% DCM; 100 mL of 0.5% MeOH+99.5% DCM; and 300 mL of 10% MeOH+90% DCM) to give the pure product (900 mg, 49% yield).

[0261] Step 3: Synthesis of compound 81-5 81-4 (716.7 mg, 1.9 mmol), DCC (399.1 mg, 1.9 mmol), DMAP (118.1 mg, 0.97 mmol), and DCM (3 mL) were added to a 25 mL flask, and the mixture was stirred at room temperature for 30 min. 81-3 (500 mg, 0.76 mmol) was then added dropwise, and the mixture was stirred at room temperature overnight. The extent of the reaction was monitored by TLC (20% EtOAc / n-heptane). After completion of the reaction, the mixture was filtered, and the filtrate was washed with 5 mL of water and 5 mL of saturated brine, dried over anhydrous Na2SO4, and filtered. The organic phase was concentrated to give the crude product (approximately 1.2 g). The crude product was purified and separated by flash column (12 g silica gel; mobile phases were 100 mL of n-heptane; 100 mL of 1% EtOAc + 99% n-heptane; 100 mL of 2% EtOAc + 98% n-heptane; and 100 mL of 8% EtOAc + 92% n-heptane) to give the pure product (640 mg, 59% yield). 1H NMR (400 MHz, CDCl3): δ 7.65-7.63 (m, 4H), 7.43-7.34 (m, 6H), 5.17 (m, 1H), 4.07-4.04 (m, 6H), 3.73 (m, 1H), 2.69 (m, 2H), 2.44-2.41 (m, 2H), 2.32-2.28 (m, 7H), 2.22-2.16 (m, 2H), 1.68-1.55 (m, 20H), 1.45-1.36 (m, 11H), 1.31-1.21 (m, 50H), 1.05 (s, 9H), 0.89-0.85 (m, 12H).

[0262] Step 4: Synthesis of Compound 81 81-5 (520 mg, 0.46 mmol) and THF (3 mL) were added to a 25 mL flask, and the mixture was cooled to 0–5 °C in an ice bath. TBAF (364.6 mg, 1.4 mmol) was added dropwise while maintaining the temperature at 0–5 °C, and the mixture was stirred at this temperature for 3 h. The extent of the reaction was monitored by TLC (5% MeOH / DCM). After completion of the reaction, the reaction was quenched with saturated aqueous ammonium chloride solution, and the mixture was extracted with EtOAc (5 mL × 2). The organic phases were combined, dried over anhydrous Na2SO4, filtered, and concentrated to give the crude product (approximately 500 mg). The crude product was purified and separated by flash column (14 g silica gel packed with 100 mL of n-heptane; mobile phases were 100 mL of 1% EtOAc + 99% n-heptane; 100 mL of 2% EtOAc + 98% n-heptane; 100 mL of 10% EtOAc + 90% n-heptane; 100 mL of 30% EtOAc + 70% n-heptane; and 100 mL of 50% EtOAc + 50% n-heptane) to give the pure product (40 mg, 10% yield). 1 H NMR (400 MHz, CDCl3): δ 5.21 (m, 1H), 4.37-4.33 (m, 1H), 4.12-4.04 (m, 5H), 3.67 (m, 1H), 2.78 (m, 2H), 2.48 (m, 2H), 2.34-2.18 (m, 9H), 1.85 (m, 2H), 1.68-1.20 (m, 78H), 0.89-0.85 (m, 12H).

[0263] Example 2: Preparation of lipid nanoparticles TMF Preparation of lipid nanoparticles TMF1 (containing 46.3% of compound ALC-0315): 1. Compounds ALC-0315 (cationic lipid), DSPC (helper lipid), cholesterol (helper lipid), and ALC-0159 (helper lipid) were dissolved in ethanol to obtain an oil phase (ethanol phase) stock solution in which the molar ratios of the four lipids were 46.3%, 9.4%, 42.7%, and 1.6%, respectively. 2. The luciferase-expressing mRNA stock solution was diluted to 0.3 mg / mL with citrate buffer (pH 4) to obtain an aqueous phase. 3. The oil phase (ethanol phase) containing the four lipid mixture and the aqueous phase containing mRNA were quickly mixed at a volume ratio of 1:3 to prepare TMF1, with a total lipid to mRNA weight ratio of 25.7:1. The sample was then concentrated using an ultrafiltration tube with a molecular weight cutoff of 100 kDa, washed with PBS buffer (pH 7.4), filtered, and finally buffer-exchanged with 120 mg / mL sucrose in PBS buffer to obtain the final sample.

[0264] Preparation of lipid nanoparticles TMF2 (containing 46.3% compound 1): 1. Compound 1 (cationic lipid / GOLD lipid), DSPC (helper lipid), cholesterol (helper lipid), and ALC-0159 (helper lipid) were dissolved in ethanol to obtain an oil phase (ethanol phase) stock solution in which the molar ratios of the four lipids were 46.3%, 9.4%, 42.7%, and 1.6%, respectively. 2. The luciferase-expressing mRNA stock solution was diluted to 0.3 mg / mL with citrate buffer (pH 4) to obtain an aqueous phase. 3. The oil phase (ethanol phase) containing the four lipid mixture and the aqueous phase containing mRNA were quickly mixed at a volume ratio of 1:3 to prepare TMF2, with a total lipid to mRNA weight ratio of 26.8:1. The sample was then concentrated using an ultrafiltration tube with a molecular weight cutoff of 100 kDa, washed with PBS buffer (pH 7.4), filtered, and finally buffer-exchanged with 120 mg / mL sucrose in PBS buffer to obtain the final sample.

[0265] Lipid nanoparticles TMF3 to TMF13 were prepared with reference to the manufacturing method of TMF2, and the lipid composition was replaced according to the GOLD lipids and helper lipids listed in Table 2.

[0266] [Table 2]

[0267] Preparation of lipid nanoparticles TMF14 (three components; containing 46.3% Compound 1 without DSPC): 1. Compound 1 (cationic lipid / GOLD lipid), cholesterol (helper lipid), and ALC-0159 (helper lipid) were dissolved in ethanol to obtain an oil phase (ethanol phase) stock solution with the molar ratios of the three lipids being 51.1%, 47.1%, and 1.8%, respectively. 2. The luciferase-expressing mRNA stock solution was diluted to 0.3 mg / mL with citrate buffer (pH 4) to obtain an aqueous phase. 3. The oil phase (ethanol phase) containing the three lipid mixtures and the aqueous phase containing mRNA were quickly mixed at a volume ratio of 1:3 to prepare TMF14, with a total lipid to mRNA weight ratio of 23.8:1. The sample was then concentrated using an ultrafiltration tube with a molecular weight cutoff of 100 kDa, washed with PBS buffer (pH 7.4), filtered, and finally buffer-exchanged with 120 mg / mL sucrose in PBS buffer to obtain the final sample.

[0268] Preparation of lipid nanoparticles TMF15 (two components; containing 66% Compound 1 and 34% DOPE): 1. Compound 1 (cationic lipid / GOLD lipid) and DOPE (helper lipid) were dissolved in ethanol to obtain an oil phase (ethanol phase) stock solution in which the molar ratios of the two lipids were 66% and 34%, respectively. 2. The luciferase-expressing mRNA stock solution was diluted to 0.09 mg / mL with citrate buffer (pH 4) to obtain an aqueous phase. 3. The oil phase (ethanol phase) containing the mixture of two lipids and the aqueous phase containing mRNA were quickly mixed in a volume ratio of 1:3 to prepare TMF15, with a total lipid to mRNA molar ratio of 1.3:2. The sample was then concentrated using an ultrafiltration tube with a molecular weight cutoff of 100 kDa, washed with PBS buffer (pH 7.4), filtered, and finally buffer-exchanged with 120 mg of sucrose in PBS buffer to obtain the final sample.

[0269] Preparation of lipid nanoparticles TMF16 (single component; containing 100% compound 1): 1. Compound 1 (cationic lipid / GOLD lipid) was dissolved in ethanol to obtain an oil phase (ethanol phase) stock solution with a lipid molar ratio of 100%. 2. The luciferase-expressing mRNA stock solution was diluted to 0.09 mg / mL with citrate buffer (pH 4) to obtain an aqueous phase. 3. The oil phase (ethanol phase) containing one lipid mixture and the aqueous phase containing mRNA were quickly mixed in a volume ratio of 1:3 to prepare TMF16, which has a molar ratio of compound 1 to mRNA of 1.3:2. The sample was then concentrated using an ultrafiltration tube with a molecular weight cutoff of 100 kDa, washed with PBS buffer (pH 7.4), filtered, and finally the filtrate was buffer-exchanged with 120 mg of sucrose in PBS buffer to obtain the final sample.

[0270] Lipid nanoparticles TMF17 to TMF53 were prepared with reference to the manufacturing method of TMF2, and the lipid composition was replaced with the cationic lipid and helper lipid listed in Table 3.

[0271] [Table 3] TIFF2025538636000091.tif224149 TIFF2025538636000092.tif123149

[0272] Example 3: Detection of lipid nanoparticles The size and polydispersity index (PDI) of the lipid nanoparticles were measured by dynamic light scattering (DLS) technique using a NanoBrook 90plus PLAS (Brookhaven Instruments, USA) at a side scattering angle of 90°. The measurement results are shown in Table 4.

[0273] [Table 4] TIFF2025538636000094.tif223149 TIFF2025538636000095.tif222149 TIFF2025538636000096.tif153149

[0274] Example 4: Detection of organ distribution of delivery systems The liposomes prepared in Example 2 were injected into 6-8 week-old female ICR mice at a dose of 0.5 mg / kg through the medial canthal venous plexus of the fundus. Four hours after administration, 15 mg / mL of D-luciferin potassium salt was intraperitoneally injected at a dose of 150 mg / kg. Ten minutes after injection of the luciferase substrate, the mice were placed in an in vivo imaging system (IVIS Lumina XRMS Series III, PerkinElmer) and observed for in vivo fluorescence intensity and distribution. The mice were then sacrificed, and organs (heart, liver, spleen, lungs, and kidneys) were isolated for ex vivo imaging to observe the fluorescence intensity and distribution in different organs. The organ distribution of Luc-mRNA delivered by representative lipid compounds is shown in Table 5. As shown in Table 5, four hours after intravenous administration of lipid nanoparticles TMF10, TMF21, TMF19, TMF26, and TMF29 containing Compound 1, Compound 25, Compound 19, Compound 35, and Compound 38, respectively, to mice, the ratio of fluorescence intensity in the spleen to the liver (spleen / liver) exceeded 60, with TMF29 having the highest ratio at 69.5. The fluorescence intensity in different organs indicates the delivery efficiency of the corresponding delivery system in different organs. The lipid nanoparticles of the present application can successfully deliver nucleic acid molecules to the spleen, enabling their expression, with the delivery efficiency to the spleen being significantly higher than that to the liver and other tissues / organs.

[0275] [Table 5] TIFF2025538636000098.tif189149

[0276] The present application has been described in detail above in order to enable those skilled in the art to understand and implement the structure and content of the present application, but this description does not limit the protection scope of the present application, and any equivalent changes or modifications made in accordance with the spirit of the present application shall fall within the protection scope of the present application.

[0277] Industrial Applicability The cationic lipid compounds provided by the present application can be applied to lipid nanoparticles, which can target different tissues and organs for drug delivery, and the lipid nanoparticles may further comprise at least one helper lipid. The cationic lipid compounds or lipid nanoparticle compositions containing the same can specifically deliver prophylactic / therapeutic agents (especially nucleic acid components) to target organs.

Claims

1. A compound of formula (I), 【Chemistry 1】 or a pharmaceutically acceptable salt, prodrug or stereoisomer thereof, wherein X １ and X ２ are each independently —OC(═O)—, —C(═O)O—, —NHC(═O)—, or —C(═O)NH—, Y １ and Y ２ are each independently —OC(═O)—, —C(═O)O—, or —NR ５ C(=O)-, -C(=O)NR ５ -bond, optionally substituted C1-C8 alkylene, or optionally substituted C2-C8 alkenylene, where R ５ is selected from hydrogen or a straight or branched chain C1-C8 hydrocarbyl; L １ and L ２ each independently represents a bond or an optionally substituted C1-C10 alkylene; R ３ and R ４ are each independently methyl, ethyl, propyl, hydroxymethyl, hydroxyethyl, hydroxypropyl, hydroxybutyl, 【Chemistry 2】 or R ３ and R ４ together with the nitrogen attached thereto form a cyclic moiety, where R ３ or R ４ is selected from the group containing hydrogen or deuterium, and the cyclic moiety is a 4- to 8-membered heterocycloalkyl selected from azetidin-1-yl, pyrrolidinyl, piperidin-1-yl, 4-hydroxypiperidin-1-yl, azepan-1-yl, morpholinyl, and 4-acetylpiperazin-1-yl; R １ and R ２ are each independently H or deuterium or a straight or branched chain C1-C8 hydrocarbyl; m is an integer from 1 to 13; n is an integer from 1 to 13; A compound or a pharmaceutically acceptable salt, prodrug or stereoisomer thereof.

2. 2. The compound of claim 1, or a pharmaceutically acceptable salt, prodrug, or stereoisomer thereof, wherein the compound is selected from the compounds shown in Table 1.

3. 10. A lipid nanoparticle comprising the compound of claim 1 or a pharmaceutically acceptable salt, prodrug or stereoisomer thereof.

4. The lipid nanoparticle of claim 3, further comprising a helper lipid, wherein the helper lipid is selected from one or more of a phospholipid, a steroid, a polymer-conjugated lipid, and a modifiable lipid.

5. The lipid nanoparticle of claim 4, wherein the phospholipid is selected from any one of DOPE, DSPC, DPPC, DMPC, DOPC, POPC, and SM, or a combination thereof.

6. The lipid nanoparticle of claim 4, wherein the steroid is selected from one or more of cholesterol, sitosterol, stigmasterol, and ergosterol, and more preferably, the steroid is cholesterol and sitosterol.

7. The lipid nanoparticle of claim 4, wherein the polymer-conjugated lipid is selected from lipids conjugated to polyethylene glycol, polylactic acid, polyamide, cationic polymer, polysarcosine (pSar), polylactic-co-glycolic acid (PLGA), polyamino acid, polypeptide or polypeptoid.

8. The lipid nanoparticle of claim 7, wherein the polymer-conjugated lipid is a polyethylene glycol-conjugated lipid selected from one or more of ALC-0159, PEG1000-DMG, PEG5000-DMG, PEG2000-DMG, and PEG2000-DSPE.

9. The lipid nanoparticle according to claim 4, wherein the molar ratio of the compound according to claim 1 or a pharmaceutically acceptable salt, prodrug or stereoisomer thereof to the helper lipid is 1: (0.5 to 2), preferably 1: (0.6 to 1.5), more preferably 1: (0.8 to 1.2).

10. 10. Use of lipid nanoparticles according to any one of claims 3 to 9 in targeted delivery, preferably wherein the target organ of said targeted delivery is lung, heart, brain, spleen, lymph node, bone, skeletal muscle, stomach, small intestine, large intestine / colorectum, kidney, bladder, breast, testis, ovary, uterus, spleen, thymus, brainstem, cerebellum, spinal cord, eye, ear, tongue or skin, preferably wherein the target organ is the spleen.

11. A composition comprising the lipid nanoparticles according to any one of claims 3 to 9 and a therapeutic / prophylactic agent, wherein preferably the therapeutic / prophylactic agent is a nucleic acid, and more preferably the nucleic acid is selected from single-stranded DNA, double-stranded DNA, single-stranded RNA, double-stranded RNA, short isomers, plasmid DNA, complementary DNA / cDNA, antisense oligonucleotides / ASO, small interfering nucleic acids / siRNA, small activating nucleic acids / saRNA, asymmetric interfering nucleic acids / aiRNA, micronucleic acids / miRNA, miRNA inhibitors (agomirs or antagomirs), dicer substrate nucleic acids, short hairpin nucleic acids (shRNA), transfer RNA (tRNA), messenger RNA / mRNA, circular RNA / circRNA, self-amplifying mRNA / saRNA, or an aptamer.

12. The composition of claim 11 , wherein the nucleic acid comprises natural nucleotides, nucleotide mimetics, functional analogs, or chemically modified nucleotides.

13. 13. The composition according to claim 11, wherein the average particle size is from 90 nm to 600 nm, preferably from 200 nm to 400 nm, more preferably from 200 nm to 300 nm.

14. 13. The composition according to any one of claims 11 and 12, having a polydispersity index (PDI) of 0.001 to 0.5, preferably 0.001 to 0.45, more preferably 0.001 to 0.

4.

15. 13. The composition according to claim 11, wherein the mass ratio of the lipid nanoparticles to the therapeutic / prophylactic agent in the composition is from 10:1 to 100:1, preferably from 20:1 to 50:1, and more preferably from 20:1 to 30:

1.

16. A medicament comprising the composition according to any one of claims 11 to 15 and pharmaceutically acceptable auxiliary substances.

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