Phosphorylcholine structure compound and preparation method thereof

Abstract

The invention relates to a phosphatidylcholine structure compound and a preparation method thereof, and the preparation method of the phosphatidylcholine structure compound comprises the following steps: taking hydroxyethyl methylacrylate and beta-mercaptoethylamine as raw materials, and reacting at room temperature to obtain an intermediate M1; carrying out addition reaction on the intermediate M1 and acryloyl ester to obtain an intermediate M2; the intermediate M2 and 2-chloro-2-oxo-1, 3, 2-dioxaphospholane are subjected to a reaction, and an intermediate M3 is obtained; and reacting the intermediate M3 with chain or cyclic tertiary amine to obtain the phosphatidylcholine compound. The phosphatidylcholine compound obtained by the method disclosed by the invention is simple in reaction and mild in condition, can be combined with numerous compound fragments on the market, and is rich and diversified in products. The phosphatidylcholine compound is used for delivery of wrapped nucleic acid drugs and can be selectively delivered to the spleen part.

Description

[0001] This invention is a divisional application of application number 202410979020.1, filed on July 22, 2024, of the invention type, entitled "A phosphatidylcholine structural compound and its preparation method". Technical Field

[0002] This invention belongs to the field of medicinal chemistry technology, specifically relating to a series of phosphatidylcholine structural compounds, preparation methods, and the application of the phosphatidylcholine compounds in targeted nucleic acid drug delivery. Background Technology

[0003] Phospholipids are an important component of biological membranes. Their inherent hydrophilic / lipophilic properties allow them to spontaneously form closed bilayers in water, becoming the biological membrane framework. Utilizing this property, in 1971, Lehman et al. in the UK began using them as liposomes as drug carriers. Since then, liposomes have attracted worldwide attention as a novel dosage form for targeted drug delivery systems. These liposomes have advantages such as low toxicity to the body, good similarity and tissue compatibility between their lipid bilayer and biological membranes, and easy absorption by tissues.

[0004] Phospholipids play a crucial role in liposomes, but the application of natural phospholipids is limited due to their susceptibility to oxidation and poor stability. Synthetic phospholipids possess strong antioxidant capabilities, resulting in more ideal liposome preparations and holding significant potential for drug delivery. Phosphatidylcholine, an amphoteric molecule composed of a hydrophilic head and a hydrophobic tail, is a type of phospholipid with a choline group inserted at the head. Lipid nanoparticles are one of the main carriers for effectively delivering nucleic acid drugs, protecting them from rapid degradation in vivo, prolonging their circulation time, and enabling more efficient delivery to target cells. Among these, helper phospholipids play a key role in nucleic acid drug delivery. Therefore, developing novel helper phospholipids is also of great importance in nucleic acid drug research. Currently, the two most common choline compounds are distearylphosphatidylcholine (DSPC) and dipalmitoylphosphatidylcholine (DPPC).

[0005] While synthetic phospholipids overcome the drawbacks of natural phospholipids, they also suffer from disadvantages such as rapid metabolism in vivo and short duration of action. The derivatization of synthetic phospholipids has promoted advancements in liposome technology for targeted and long-circulation applications. Given the widespread use of DSPC and DPPC in liposome research and the resulting increase in market demand, developing novel synthetic methods for phosphatidylcholine structures is of great significance.

[0006] Patent (CN111057099A) discloses a dipalmitoylphosphatidylcholine (DPPC) and its preparation method. This method uses the condensation of GPC and palmitic acid to prepare DPPC. This method only targets the preparation of DPPC and is unlikely to enrich the product library.

[0007] Patent (CN114213458A) discloses a cyclobutamine-terminated distearate phosphatidylcholine and its preparation method. This method is limited to the synthesis of cyclobutamine-terminated compounds, which has significant limitations and a narrow scope.

[0008] Therefore, there is an urgent need in this field to develop a series of novel methods for preparing phosphatidylcholine, so as to better meet the market demand for phosphatidylcholine compounds. Summary of the Invention

[0009] The purpose of this invention is to provide a series of phosphatidylcholine structural compounds, their preparation methods, and applications. These novel phosphatidylcholines exhibit mild reaction conditions, simple operation, and easy purification during synthesis. Furthermore, they have abundant synthetic intermediates and can bind to various chain-like or cyclic tertiary amines and ester groups, enabling the differentiated synthesis of various types of phosphatidylcholine compounds and enriching the phosphatidylcholine product library. To address the aforementioned technical problems, this invention provides the following technical solutions:

[0010] This invention provides a phosphatidylcholine compound with the structure of formula (I) or (II), a pharmaceutically acceptable salt of the compound, a tautomer or a stereoisomer.

[0011]

[0012] Where X is selected from -O- or -S-;

[0013] Ra is selected from C 3-8 The alkylene group, wherein the carbon atoms in the alkylene group are optionally replaced by 1-3 -O-, -S-, -NH- groups, provided that the -O-, -S-, -NH- groups are not directly bonded to the X group or the R1R2N- group, and that the -O-, -S-, -NH- groups are not directly bonded to each other; optionally, one or more hydrogen atoms in Ra are replaced by Rb, wherein Rb is selected from C 1-8 alkyl.

[0014] It is a chain-like or cyclic quaternary amine, and R3, R4, and R5 are each independently selected from C. 1-8 The branched or straight-chain alkyl group, or one of the N atom (R3, R4, R5) together with its adjacent counterpart forms a 5- or 6-membered heterocycle; optionally, the 5- or 6-membered heterocycle is replaced by one or more R6 atoms, wherein the R6 is selected from C 1-6 Branched or straight-chain alkyl groups.

[0015] R1 and R2 are each independently selected from -H and C. 2-8 Straight-chain or branched alkyl groups, wherein C 2-8 The methylene groups contained in the straight-chain or branched alkyl groups are optionally replaced by -S-, -O-, or -NH-, and one or more hydrogens in R1 and R2 are optionally replaced by -COOR7, -OCOR7, -COSR7, -SCOR7, -CONR7R8, or -NR7COR8, wherein R7 and R8 are each independently selected from -H, C 1-20 Branched or straight-chain alkyl groups, C 1-20 Branched or straight-chain alkenyl groups, C 1-20 It has a branched or straight-chain alkynyl group, and R1 and R2 are not both -H.

[0016] Preferably, R1 and R2 are selected from -H or groups with the structure described below, and R1 and R2 are not both -H.

[0017]

[0018]

[0019] Further preferred, It has the following structure:

[0020]

[0021] This invention provides a phosphatidylcholine compound with the structure of formula (Ia) and (IIa), a pharmaceutically acceptable salt of the compound, a tautomer or a stereoisomer, wherein the definitions of R1, R2, R3, R4, R5, X and Rb are as described above.

[0022]

[0023] More preferably, the compound of formula (I) of the present invention has the following structure:

[0024]

[0025]

[0026]

[0027]

[0028] Furthermore, the present invention also provides the use of a phosphatidylcholine compound of formula (I), a pharmaceutically acceptable salt of the compound, a tautomer or a stereoisomer, as a drug carrier, preferably, the drug being selected from nucleic acid drugs.

[0029] Furthermore, the present invention also provides a method for preparing phosphatidylcholine compounds with the structure of formula (Ia), pharmaceutically acceptable salts of said compounds, tautomers or stereoisomers, the method comprising the following steps:

[0030] S1. Using hydroxyethyl methacrylate and β-mercaptoethylamine as raw materials, intermediate M1 is obtained by reacting in a solvent at room temperature; the reaction formula is as follows:

[0031]

[0032] S2. Intermediate M1 and An addition or substitution reaction is carried out to give intermediate M2; the reaction formula is:

[0033] R9 is selected from C. 0-6 The R9 is a straight-chain or branched alkyl group, wherein the methylene group in R9 is optionally replaced by -S-, -O-, or -NH-, and one or more hydrogens in R9 are optionally replaced by -COOR7, -OCOR7, -CONR7R8, -NR7COR8, -COSR7, or -SCOR7, wherein R7 and R8 are each independently selected from -H and C. 1-20 Branched or straight-chain alkyl groups, C 1-20 Branched or straight-chain alkenyl groups, C 1-20 Branched or straight-chain alkynyl groups,

[0034] S3. Intermediate M2 reacts with 2-chloro-2-oxo-1,3,2-dioxophosphazenecyclopentane to give intermediate M3; the reaction formula is:

[0035]

[0036] S4. Intermediate M3 and The reaction yields the phosphatidylcholine compound represented by formula Ia; the reaction formula is:

[0037]

[0038] The definitions of R1, R2, R3, R4, and R5 are as described above.

[0039] Preferably, in step S1, the molar ratio of SM1 and SM2 should be between 1:1 and 1.2, with the optimal molar ratio being 1:1.

[0040] Preferably, in step S1, the reaction temperature should be between 10 and 30°C, and the reaction time should be between 3 and 6 hours.

[0041] Preferably, in step S1, the solvent is anhydrous ethanol, and the volume ratio of hydroxyethyl methacrylate to the solvent is 1:5-10 (g / ml).

[0042] Preferably, in step S1, the post-processing operation is as follows: the reaction solution is concentrated under reduced pressure to remove the solvent, resulting in a colorless oily crude product, which is directly used in the next reaction without further purification.

[0043] Preferably, in step S2, the molar ratio of M1 to SM3 should be between 1:2.2 and 3, with the optimal molar ratio being 1:2.5.

[0044] Preferably, in step S2, a solvent-free reaction is used, the reaction temperature should be between 70 and 80°C, and the time should be between 40 and 60 hours.

[0045] Preferably, in step S2, 0.05 equivalents of BHT should be added as a reaction stabilizer (the reaction stabilizer can prevent polymerization from occurring during the reaction process).

[0046] Preferably, in step S2, 0.05 equivalent of catalyst is added, and the catalyst is preferably acetic acid or boric acid.

[0047] Preferably, in step S3, the reaction occurs under the action of the acid-binding agent, and the molar ratio of M2 to SM4 and the acid-binding agent is 1:2:2.5. The acid-binding agent is selected from organic or inorganic bases. Preferably, the organic base is selected from, but is not limited to, one or more of triethylamine, diethylisopropylamine, pyridine, DMAP, and DBU, and the inorganic base is selected from one or more of sodium carbonate, potassium carbonate, sodium bicarbonate, and potassium bicarbonate.

[0048] Preferably, in step S3, the reaction is carried out in an organic inert aprotic solvent, preferably selected from one or more of dry dichloromethane, tetrahydrofuran, DMF, DMSO, acetone, and dioxane, with a substrate to solvent mass-volume ratio of 1:10 (g / ml).

[0049] Preferably, in step S3, the reaction temperature should be between 0 and 10°C, and the reaction time should be between 1 and 3 hours.

[0050] Preferably, in step S3, there are two post-processing operations. The first post-processing operation is as follows: Due to the poor stability of the intermediate, it cannot be purified. Therefore, the reaction solution is directly concentrated under reduced pressure to remove the solvent, yielding a milky white oily crude product. This crude product is then slurried with ethyl acetate to obtain a white suspension. The solid is removed by filtration, and the mother liquor is concentrated under reduced pressure to obtain the crude product. The crude product is directly used in the next reaction step without further purification. The product has poor stability and therefore cannot be characterized. If the acid-binding agent used in step S3 is the same as the tertiary amine used in step S4, the second post-processing operation is adopted: the reaction solution does not require further post-processing and is directly used for feeding in step S4.

[0051] Preferably, in step S4, M3 and The molar ratio is 1:1.5 to 5, preferably 1:3.

[0052] Preferably, in step S4, the reaction is carried out in a solvent selected from one or more of dry acetonitrile, tetrahydrofuran, DMF, DMSO, acetone, and dioxane. The mass-to-volume ratio of substrate to solvent is 1:5-10 (g / ml).

[0053] Preferably, in step S4, the reaction temperature should be between 50 and 55°C, and the reaction time should be between 20 and 50 hours.

[0054] Preferably, in step S4, the post-processing operation is as follows: the reaction solution is concentrated under reduced pressure to remove the solvent and obtain a pale yellow oily crude product, which is then directly purified by column chromatography to obtain the target product.

[0055] Furthermore, the present invention provides a composition comprising a phosphatidylcholine compound with the structure of formula (I) or (II) above, a pharmaceutically acceptable salt of the compound, a tautomer or a stereoisomer.

[0056] Furthermore, the present invention provides the above-described composition, wherein the composition further comprises other lipids.

[0057] Furthermore, the present invention provides the above-described composition, wherein the other lipids are selected from... One or more of cholesterol and DMG-PEG2000.

[0058] Furthermore, the present invention provides the above-described composition, wherein the other lipids are selected from... One or more of cholesterol and DMG-PEG2000.

[0059] Furthermore, the present invention provides the above-described composition, wherein the other lipids are selected from... One or more of cholesterol and DMG-PEG2000.

[0060] Furthermore, the present invention provides the above-described composition, wherein the other lipids are selected from... One or more of cholesterol and DMG-PEG2000.

[0061] Furthermore, the present invention provides the above-described composition, wherein the other lipids are selected from... One or more of cholesterol and DMG-PEG2000.

[0062] Furthermore, the present invention provides the above composition in which the other lipids, phosphatidylcholine compounds of formula (I) or formula (II), cholesterol, and DMG-PEG2000 have a molar percentage of (40% to 60%): (5% to 15%): (22% to 54.5%): (0.5% to 1.5%).

[0063] Furthermore, the present invention provides the above-described composition, wherein the composition further comprises cholesterol and DMG-PEG2000.

[0064] The C of this invention 1-6 Or C 1-8 The alkyl group is preferably selected from methyl, ethyl, n-propyl, isopropyl, n-butyl, 1-methylpropyl, 2-methylpropyl, tert-butyl, n-pentyl, n-hexyl, n-heptyl, and n-octyl.

[0065] The C of this invention 1-20 Branched or straight-chain alkyl groups, C 1-20 Branched or straight-chain alkenyl groups, C 1-20 The number of carbon atoms in the branched or straight-chain alkynyl group is preferably 6-20, 8-20, 10-20, 12-20, 14-20, 16-20 or 18-20.

[0066] In this invention, alkyl refers to an alkane with one hydrogen atom in a corresponding number of carbon atoms in which one hydrogen atom is replaced by another group, and alkylene refers to an alkane with two hydrogen atom in a corresponding number of carbon atoms in which two hydrogen atom is replaced by another group, such as the structure of methylene being -CH2- and the structure of ethylene being -CH2CH2-.

[0067] The term "optional substitution" in this invention refers to the fact that the substitution may or may not occur.

[0068] Compared with the prior art, the present invention has the following beneficial effects:

[0069] The synthesis process of this invention is mild, simple to operate, and easy to purify.

[0070] This invention enables the differentiated synthesis of various types of phosphatidylcholine compounds, which is beneficial for enriching the phosphatidylcholine product library and providing many options and directions for the application of liposomes.

[0071] The phosphatidylcholine compounds obtained by the method of the present invention have simple reaction conditions and mild conditions, and can bind to a wide variety of commercially available compound fragments, resulting in a rich variety of products; such phosphatidylcholine compounds can be selectively delivered to the spleen site for the delivery of nucleic acid-encapsulated drugs. Attached Figure Description

[0072] Figure 1 This is a schematic diagram of the reaction route for the synthesis of the phosphatidylcholine compound of the present invention;

[0073] Figure 2 Fluci is expressed in HEK-293T cells;

[0074] Figure 3 Fluci is expressed in Huh7 cells;

[0075] Figure 4 The formulation of lipids of compounds (1) and (4) was expressed by 1LNP and SM-102LNP formulation Fluci;

[0076] Figure 5 Fluci expression levels in various organs of the formulations of lipids of compounds (1) and (4) 1LNP and SM-102LNP;

[0077] Figure 6 Fluci expression in the liver and spleen of formulation 1LNP containing compounds (2), (13), (24), (5), (16) and (27);

[0078] Figure 7 Fluci expression levels in various organs of lipid formulations of compounds (2), (5), (13) and (16) of compound (16). Detailed Implementation

[0079] To make the technical means, creative features, objectives and effects of this invention easier to understand, the invention will be further described below in conjunction with specific embodiments.

[0080] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. The terminology used herein in the specification of this invention is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. The term "and / or" as used herein includes any and all combinations of one or more of the associated listed items.

[0081] The present invention will be further described below with reference to the accompanying drawings and specific embodiments, so that those skilled in the art can better understand and implement the present invention. However, the embodiments described are not intended to limit the present invention.

[0082] Unless otherwise specified, the experimental methods used in the following examples are conventional methods, and the materials and reagents used are commercially available.

[0083] Example 1

[0084] Synthesis of a novel phosphatidylcholine compound (1)

[0085] The structural formula of the novel phosphatidylcholine compound (1) is as follows:

[0086]

[0087] Step S1:

[0088]

[0089] Under nitrogen protection and at room temperature, hydroxyethyl methacrylate (5.0 g, 1.0 eq.) and anhydrous ethanol (50 mL, 10V) were added sequentially to the reaction flask. Mercaptoethylamine (2.96 g, 1.0 eq.) was then added with stirring, and the mixture was stirred at room temperature for 3 hours. TLC analysis (PE / DCM = 1 / 1) showed that the hydroxyethyl methacrylate starting material was completely consumed. The reaction solution was concentrated under reduced pressure to remove the solvent, yielding a colorless oily crude product (8.0 g, crude product), which was directly used in the next reaction step. The synthesized intermediate compound M1 was subjected to NMR analysis, and its characterization data are as follows: 1H NMR (400MHz, Chloroform-d) δ 4.31 (ddd, J=11.7,5.3,3.8Hz,1H), 4.22–4.15 (m,1H), 3.81 (ddd, J=5.3,3.8,1.2Hz,2H), 2.90–2.64 (m,8H), 1.27 (d,J=6.7Hz,3H).

[0090] Step S2:

[0091]

[0092] At room temperature, M1 (1.0 g, 1.0 eq.), SM3 (containing 0.05% BHT stabilizer) (3.87 g, 2.5 eq.), and acetic acid (14.49 mg, 0.05 eq.) were added sequentially to a reaction flask. The mixture was stirred and heated to 70°C for 40 h. TLC (DCM / MeOH = 20 / 1) analysis showed that the starting materials were almost completely consumed. The reaction solution was directly purified by silica gel column chromatography (DCM / MeOH = 1 / 0-100 / 1). 2.7 g of the target product, a pale yellow oil, was obtained, with a yield of 65.97%. The synthesized intermediate compound M2 was characterized by NMR analysis, and the data are as follows: ¹H NMR (400MHz, Chloroform-d) δ 5.43–5.27 (m, 8H), 4.36–4.28 (m, 1H), 4.23–4.17 (m, 1H), 4.06 (t, J = 6.8Hz, 4H), 3.83 (t, J = 4.6Hz, 2H), 2.86–2.57 (m, 16H), 2.44 (t, J = 7.2Hz, 4H), 2.05 (q, J = 6.8Hz, 8H), 1.62 (p, J = 6.8Hz, 4H), 1.40–1.22 (m, 37H), 0.93–0.84 (m, 6H). The synthesized intermediate compound M2 was analyzed by high-resolution mass spectrometry, and its characterization data are as follows: MS (m / z): [M+H] + =848.69978.

[0093] Step S3:

[0094]

[0095] M2 (500 mg, 1.0 eq.) and anhydrous DCM (5 mL, 10 V) were added sequentially to the reaction flask. The mixture was cooled to 4 °C in an ice bath, and 2-chloro-2-oxo-1,3,2-dioxophosphazenecyclopentane (167.95 mg, 2.0 eq.) was added. After the addition was complete, the mixture was stirred for 5 minutes. Triethylamine (149.10 mg, 2.5 eq.) was then slowly added dropwise. One hour after the addition was complete, TLC (PE / EA = 1 / 1) showed that the starting material had been completely consumed, and the reaction was considered complete. The reaction solution was concentrated under reduced pressure to remove DCM, and EA (5 mL, 10 V) was added. The mixture was filtered, and the mother liquor was collected and concentrated under reduced pressure to obtain 600 mg of a colorless oily product. The crude product had poor stability and was not characterized; it was used directly in the next reaction step.

[0096] Step S4:

[0097]

[0098] Under nitrogen protection and at room temperature, M3 (150 mg crude product, 1.0 eq.), anhydrous acetonitrile (0.75 mL, 5V), and trimethylamine (2 M in THF) (27.87 mg, 3.0 eq.) were added sequentially to the reaction flask. The mixture was slowly heated to 50 °C and stirred for 30 h. TLC (PE / EA = 1 / 1) analysis showed complete consumption of the starting materials. The reaction solution was directly purified by silica gel column chromatography (DCM / MeOH = 1 / 0-5 / 1). 33 mg of the target product, a white waxy substance, was obtained, with a yield of 20.72%. The synthesized product was characterized by NMR analysis, and the data are as follows: ¹H NMR (400MHz, Chloroform-d) δ 5.28 (dtt, J=17.8,11.4,5.1Hz,8H), 4.23 (dt, J=10.2,5.2Hz,2H), 3.99 (t, J=6.8Hz,6H), 3.92–3.83 (m,2H), 3.66 (t, J=4.2Hz,2H), 2.92–2.37 (m,19H), 1.98 (q, J=6.8Hz,8H), 1.54 (q, J=6.9Hz,4H), 1.34–1.11 (m,39H), 0.82 (t, J=6.7Hz,6H). The synthesized product was also characterized by high-resolution mass spectrometry analysis, and the data are as follows: MS (m / z): [M+H] + =1013.67236.

[0099] Example 2 Synthesis of a novel phosphatidylcholine compound (2)

[0100] The structural formula of the novel phosphatidylcholine compound (2) is as follows:

[0101]

[0102] Step S4:

[0103]

[0104] Under nitrogen protection and at room temperature, M3 (600 mg crude product, 1.0 eq.), anhydrous acetonitrile (3.0 mL, 5V), and triethylamine (127.24 mg, 2.0 eq.) were added sequentially to the reaction flask. The mixture was slowly heated to 50 °C and stirred for 30 h. TLC (PE / EA = 1 / 1) analysis showed complete consumption of the starting materials. The reaction solution was directly purified by silica gel column chromatography (DCM / MeOH = 1 / 0-5 / 1). 148 mg of the target product, a white waxy substance, was obtained, with a yield of 22.30%. The synthesized product was characterized by NMR analysis, and the data are as follows: ¹H NMR (400MHz, Chloroform-d) δ 5.44–5.26 (m, 8H), 4.29 (t, J = 4.8Hz, 2H), 4.05 (t, J = 6.8Hz, 6H), 3.95 (q, J = 7.1, 5.1Hz, 2H), 3.73 (t, J = 4.1Hz, 2H), 2.92–2.58 (m, 15H), 2.50 (t, J = 7.2Hz, 4H), 2.05 (q, J = 6.8Hz, 8H), 1.62 (p, J = 6.7Hz, 4H), 1.40–1.19 (m, 42H), 0.93–0.77 (m, 6H). The synthesized product was also characterized by high-resolution mass spectrometry analysis, and the data are as follows: MS (m / z): [M+H] + =1055.75450.

[0105] Example 3 Synthesis of a novel phosphatidylcholine compound (3)

[0106] The structural formula of the novel phosphatidylcholine compound (3) is as follows:

[0107]

[0108] Step S4:

[0109]

[0110] Under nitrogen protection and at room temperature, M3 (150 mg crude product, 1.0 eq.), anhydrous acetonitrile (0.75 mL, 5V), and DIPEA (60.94 mg, 3.0 eq.) were added sequentially to the reaction flask. The mixture was slowly heated to 50 °C and stirred for 30 h. TLC (PE / EA = 1 / 1) analysis showed complete consumption of the starting materials. The reaction solution was directly purified by silica gel column chromatography (DCM / MeOH = 1 / 0-5 / 1). 63 mg of the target product, a white waxy substance, was obtained, with a yield of 36.99%. The synthesized product was subjected to NMR analysis, and its characterization data are as follows: ¹H NMR (400MHz, Chloroform-d) δ 5.44–5.28 (m, 8H), 4.40–4.22 (m, 2H), 4.12–3.91 (m, 8H), 3.74 (t, J = 4.1Hz, 2H), 3.00–2.50 (m, 20H), 2.05 (q, J = 6.8Hz, 9H), 1.61 (q, J = 7.0Hz, 5H), 1.31 (ddt, J = 22.4, 20.0, 6.5Hz, 47H), 0.94–0.76 (m, 12H).

[0111] Example 4 Synthesis of a novel phosphatidylcholine compound (4)

[0112] The structural formula of the novel phosphatidylcholine compound (4) is as follows:

[0113]

[0114] Step S4:

[0115]

[0116] Under nitrogen protection and at room temperature, M3 (150 mg crude product, 1.0 eq.), anhydrous acetonitrile (0.75 mL, 5V), and N,N-diethylmethylamine (46.24 mg, 3.0 eq.) were added sequentially to the reaction flask. The mixture was slowly heated to 50 °C and stirred for 30 h. TLC (PE / EA = 1 / 1) analysis showed complete consumption of the starting materials. The reaction solution was directly purified by silica gel column chromatography (DCM / MeOH = 1 / 0-5 / 1). 60 mg of the target product, a white waxy substance, was obtained, with a yield of 32.58%. The synthesized product was characterized by NMR analysis, and the data are as follows: ¹H NMR (400MHz, Chloroform-d) δ 5.45–5.26 (m, 8H), 4.41–4.22 (m, 2H), 4.13–3.92 (m, 8H), 3.74 (t, J = 4.2Hz, 2H), 2.99–2.51 (m, 20H), 2.05 (q, J = 6.9Hz, 9H), 1.62 (t, J = 7.0Hz, 4H), 1.39–1.20 (m, 42H), 0.89 (t, J = 6.8Hz, 6H). The synthesized product was also characterized by high-resolution mass spectrometry analysis, and the data are as follows: MS (m / z): [M+H] + =1041.80881.

[0117] Example 5 Synthesis of a novel phosphatidylcholine compound (5)

[0118] The structural formula of the novel phosphatidylcholine compound (5) is as follows:

[0119]

[0120] Step S4:

[0121]

[0122] Under nitrogen protection and at room temperature, M3 (150 mg crude product, 1.0 eq.), anhydrous acetonitrile (0.75 mL, 5V), and N-methyldipropylamine (54.33 mg, 3.0 eq.) were added sequentially to the reaction flask. The mixture was slowly heated to 50 °C and stirred for 30 h. TLC (PE / EA = 1 / 1) analysis showed complete consumption of the starting materials. The reaction solution was directly purified by silica gel column chromatography (DCM / MeOH = 1 / 0-5 / 1). 58 mg of the target product, a white waxy substance, was obtained, with a yield of 34.50%. The synthesized product was subjected to NMR analysis, and its characterization data are as follows: ¹H NMR (400MHz, Chloroform-d) δ 5.28 (dtt, J=17.8,11.4,5.1Hz,8H), 4.23 (dt, J=10.2,5.2Hz,2H), 3.99 (t, J=6.8Hz,6H), 3.92–3.83 (m,2H), 3.66 (t, J=4.2Hz,2H), 2.92–2.37 (m,19H), 1.98 (q, J=6.8Hz,8H), 1.54 (q, J=6.9Hz,4H), 1.34–1.11 (m,39H), 0.82 (t, J=6.7Hz,6H).

[0123] The synthesized product was analyzed by high-resolution mass spectrometry, and the characterization data are as follows:

[0124] MS(m / z):[M+H] + =1069.77214.

[0125] Example 6 Synthesis of a novel phosphatidylcholine compound (6)

[0126] The structural formula of the novel phosphatidylcholine compound (6) is as follows:

[0127]

[0128] Step S4:

[0129]

[0130] Under nitrogen protection and at room temperature, M3 (150 mg crude product, 1.0 eq.), anhydrous acetonitrile (0.75 mL, 5V), and N,N-dimethylbutylamine (56.78 mg, 3.0 eq.) were added sequentially to the reaction flask. The mixture was slowly heated to 50 °C and stirred for 30 h. TLC (PE / EA = 1 / 1) analysis showed complete consumption of the starting materials. The reaction solution was directly purified by silica gel column chromatography (DCM / MeOH = 1 / 0-5 / 1). 63 mg of the target product, a white waxy substance, was obtained, with a yield of 37.94%.

[0131] The synthesized product underwent NMR analysis, and the characterization data are as follows:

[0132] 1H NMR(400MHz,Chloroform-d)δ5.35(tdt,J=10.7,8.4,5.1Hz,8H),4.30(d,J=6.0Hz,2H),4.07(q,J=6.6Hz,6H),3.95(p,J=4.6,4.2Hz,2 H),3.73(t,J=4.3Hz,2H),2.96–2.40(m,19H),2.05(q,J=6.8Hz,8H),1.62(t,J=7.0Hz,4H),1.40–1.18(m,38H),0.89(t,J=6.8Hz,6H).

[0133] The synthesized product was analyzed by high-resolution mass spectrometry, and the characterization data are as follows:

[0134] MS(m / z):[M+H] + =1055.76551.

[0135] Example 7 Synthesis of a novel phosphatidylcholine compound (7)

[0136] The structural formula of the novel phosphatidylcholine compound (7) is as follows:

[0137]

[0138] Step S4:

[0139]

[0140] Under nitrogen protection and at room temperature, M3 (150 mg crude product, 1.0 eq.), anhydrous acetonitrile (0.75 mL, 5V), and N,N-dimethyl-1-octylamine (78.00 mg, 3.0 eq.) were added sequentially to the reaction flask. The mixture was slowly heated to 50 °C and stirred for 30 h. TLC (PE / EA = 1 / 1) analysis showed complete consumption of the starting materials. The reaction solution was directly purified by silica gel column chromatography (DCM / MeOH = 1 / 0-5 / 1). 78 mg of the target product, a white waxy substance, was obtained, with a yield of 44.64%.

[0141] The synthesized product underwent NMR analysis, and the characterization data are as follows:

[0142] 1H NMR(400MHz,Chloroform-d)δ5.35(tdd,J=11.1,6.4,3.5Hz,8H),4.30(d,J=6.6Hz,2H),4.00(dt,J=43.6,7.3Hz,8H),3.73 (t,J=4.1Hz,2H),2.97–2.43(m,20H),2.05(q,J=6.8Hz,8H),1.62(t,J=6.9Hz,4H),1.40–1.15(m,46H),0.92–0.77(m,12H).

[0143] The synthesized product was analyzed by high-resolution mass spectrometry, and the characterization data are as follows:

[0144] MS(m / z):[M+H] + =1111.80362.

[0145] Example 8 Synthesis of a novel phosphatidylcholine compound (8)

[0146] The structural formula of the novel phosphatidylcholine compound (8) is as follows:

[0147]

[0148] Step S4:

[0149]

[0150] Under nitrogen protection and at room temperature, M3 (150 mg crude product, 1.0 eq.), anhydrous acetonitrile (0.75 mL, 5V), and 1-methylpyrrolidone (40.92 mg, 3.0 eq.) were added sequentially to the reaction flask. The mixture was slowly heated to 50 °C and stirred for 30 h. TLC (PE / EA = 1 / 1) analysis showed complete consumption of the starting materials. The reaction solution was directly purified by silica gel column chromatography (DCM / MeOH = 1 / 0-5 / 1). 54.8 mg of the target product, a white waxy substance, was obtained, with a yield of 32.91%.

[0151] The synthesized product underwent NMR analysis, and the characterization data are as follows:

[0152] 1H NMR(400MHz,Chloroform-d)δ5.35(tdd,J=11.3,6.5,3.5Hz,8H),4.40–4.23(m,2H),4.05(td,J=6.8,4.1Hz,6H),3.97–3.68(m,4H),3.24(s,1H), 2.94–2.41(m,21H),2.35–2.16(m,1H),2.05(q,J=6.8Hz,8H),1.62(p,J= 6.8Hz, 4H), 1.31 (ddt, J = 22.6, 19.5, 6.6Hz, 38H), 0.89 (t, J = 6.7Hz, 6H).

[0153] The synthesized product was analyzed by high-resolution mass spectrometry, and the characterization data are as follows:

[0154] MS(m / z):[M+H] + =1039.74359.

[0155] Example 9 Synthesis of a novel phosphatidylcholine compound (9)

[0156] The structural formula of the novel phosphatidylcholine compound (9) is as follows:

[0157]

[0158] Step S4:

[0159]

[0160] Under nitrogen protection and at room temperature, M3 (150 mg crude product, 1.0 eq.), anhydrous acetonitrile (0.75 mL, 5V), and N-methylpiperidine (46.76 mg, 3.0 eq.) were added sequentially to the reaction flask. The mixture was slowly heated to 50 °C and stirred for 30 h. TLC (PE / EA = 1 / 1) analysis showed complete consumption of the starting materials. The reaction solution was directly purified by silica gel column chromatography (DCM / MeOH = 1 / 0-5 / 1). 42 mg of the target product, a white waxy substance, was obtained, with a yield of 25.36%.

[0161] The synthesized product underwent NMR analysis, and the characterization data are as follows:

[0162] 1H NMR(400MHz,Chloroform-d)δ5.35(tdd,J=11.1,6.4,3.5Hz,7H),4.40–4.20(m,2H),4.13–3.92(m,7H),3.74(t,J=4. 2Hz, 2H), 3.02–2.53 (m, 19H), 2.05 (q, J = 6.9Hz, 8H), 1.62 (p, J = 6.8Hz, 4H), 1.40–1.21 (m, 38H), 0.89 (t, J = 6.7Hz, 6H).

[0163] The synthesized product was analyzed by high-resolution mass spectrometry, and the characterization data are as follows:

[0164] MS(m / z):[M+H] + =1053.75037.

[0165] Example 10 Synthesis of a novel phosphatidylcholine compound (10)

[0166] The structural formula of the novel phosphatidylcholine compound (10) is as follows:

[0167]

[0168] Step S4:

[0169]

[0170] Under nitrogen protection and at room temperature, M3 (150 mg crude product, 1.0 eq.), anhydrous acetonitrile (0.75 mL, 5V), and N-ethylpiperidine (54.17 mg, 3.0 eq.) were added sequentially to the reaction flask. The mixture was slowly heated to 50 °C and stirred for 30 h. TLC (PE / EA = 1 / 1) analysis showed complete consumption of the starting materials. The reaction solution was directly purified by silica gel column chromatography (DCM / MeOH = 1 / 0-5 / 1). 48 mg of the target product, a white waxy substance, was obtained, with a yield of 28.19%.

[0171] The synthesized product underwent NMR analysis, and the characterization data are as follows:

[0172] 1H NMR(400MHz,Chloroform-d)δ5.28(tdd,J=11.6,10.3,5.8,3.2Hz,8H),4.31–4.15(m,2H),3.99(t,J=6.8Hz,6H),3.93–3.84(m,2H),3.66( t,J=4.2Hz,2H),3.41(s,4H),2.89–2.40(m,21H),1.98(q,J=6.9Hz,9H),1.55(t,J=7.0Hz,4H),1.34–1.13(m,41H),0.82(t,J=6.7Hz,6H).

[0173] The synthesized product was analyzed by high-resolution mass spectrometry, and the characterization data are as follows:

[0174] MS(m / z):[M+H] + =1067.74502.

[0175] Example 11 Synthesis of a novel phosphatidylcholine compound (11)

[0176] The structural formula of the novel phosphatidylcholine compound (11) is as follows:

[0177]

[0178] Step S4:

[0179]

[0180] Under nitrogen protection and at room temperature, M3 (150 mg crude product, 1.0 eq.), anhydrous acetonitrile (0.75 mL, 5V), and N,N-dimethylcyclohexylamine (59.99 mg, 3.0 eq.) were added sequentially to the reaction flask. The mixture was slowly heated to 50 °C and stirred for 30 h. TLC (PE / EA = 1 / 1) analysis showed complete consumption of the starting materials. The reaction solution was directly purified by silica gel column chromatography (DCM / MeOH = 1 / 0-5 / 1). 42 mg of the target product, a white waxy substance, was obtained, with a yield of 24.71%.

[0181] The synthesized product underwent NMR analysis, and the characterization data are as follows:

[0182] 1H NMR(400MHz,Chloroform-d)δ5.45–5.24(m,8H),4.37(s,1H),4.25(d,J=12.5Hz,1H),4.15–3.92(m,8H),3.75(d,J= 4.3Hz, 2H), 3.04–2.54 (m, 19H), 2.05 (q, J = 6.9Hz, 9H), 1.62 (t, J = 6.9Hz, 4H), 1.39–1.20 (m, 47H), 0.91–0.81 (m, 9H).

[0183] The synthesized product was analyzed by high-resolution mass spectrometry, and the characterization data are as follows:

[0184] MS(m / z):[M+H] + =1081.76979.

[0185] Example 12 Synthesis of a novel phosphatidylcholine compound (12)

[0186] The structural formula of the novel phosphatidylcholine compound (12) is as follows:

[0187]

[0188] Step S2:

[0189]

[0190] M1 (1.5 g, 1.0 eq.), octadecyl acrylate (5.17 g, 2.2 eq.), BHT (79.73 mg, 0.05 eq.), and acetic acid (21.73 mg, 0.05 eq.) were added sequentially to a reaction flask at room temperature. The mixture was stirred and heated to 70 °C for 40 h. TLC (DCM / MeOH = 20 / 1) analysis showed that the starting materials were almost completely consumed. The reaction solution was directly purified by silica gel column chromatography (DCM / MeOH = 1 / 0-100 / 1). 1.8 g of the target product, a white waxy substance, was obtained, with a yield of 29.05%.

[0191] The NMR analysis of the synthesized intermediate compound M2 yielded the following characterization data:

[0192] 1H NMR(400MHz,Chloroform-d)δ4.36–4.27(m,1H),4.24–4.15(m,1H),4.06(t,J=6.8Hz,4H),3.83(t,J=4.6Hz,2H),2.8 9–2.70(m,6H),2.69–2.54(m,6H),2.44(t,J=7.1Hz,4H),1.61(q,J=5.5,3.7Hz,7H),1.26(s,66H),0.93–0.83(m,6H).

[0193] Step S3:

[0194]

[0195] M2 (500 mg, 1.0 eq.) and anhydrous DCM (5 mL, 10 V) were added sequentially to the reaction flask. The mixture was cooled to 4 °C in an ice bath, and 2-chloro-2-oxo-1,3,2-dioxophosphazenecyclopentane (166.37 mg, 2.0 eq.) was added. After the addition was complete, the mixture was stirred for 5 minutes. Triethylamine (149.10 mg, 2.5 eq.) was then slowly added dropwise. One hour after the addition was complete, TLC (PE / EA = 1 / 1) showed that the starting material had been completely consumed, and the reaction was considered complete. The reaction solution was concentrated under reduced pressure to remove DCM, and EA (5 mL, 10 V) was added. The mixture was filtered, and the mother liquor was collected and concentrated under reduced pressure to obtain 600 mg of a colorless oily product. The crude product had poor stability and was not characterized; it was used directly in the next reaction step.

[0196] Step S4:

[0197]

[0198] Under nitrogen protection and at room temperature, M3 (150 mg crude product, 1.0 eq.), anhydrous tetrahydrofuran (0.75 mL, 5V), and trimethylamine (2 M in THF) (0.24 mL, 3.0 eq.) were added sequentially to the reaction flask. The mixture was slowly heated to 50 °C and stirred for 30 h. TLC (PE / EA = 1 / 1) analysis showed complete consumption of the starting materials. The reaction solution was directly purified by silica gel column chromatography (DCM / MeOH = 1 / 0-5 / 1). 46 mg of the target product was obtained as a white solid, with a yield of 28.89%.

[0199] The synthesized product underwent NMR analysis, and the characterization data are as follows:

[0200] 1H NMR(400MHz,Chloroform-d)δ4.37–4.22(m,2H),4.12–3.92(m,7H),3.78–3.70(m,2 H), 3.00–2.46 (m, 16H), 1.62 (p, J = 6.9Hz, 4H), 1.26 (s, 63H), 0.88 (t, J = 6.6Hz, 6H).

[0201] Example 13 Synthesis of a novel phosphatidylcholine compound (13)

[0202] The structural formula of the novel phosphatidylcholine compound (13) is as follows:

[0203]

[0204] Step S4:

[0205]

[0206] Under nitrogen protection and at room temperature, M3 (150 mg crude product, 1.0 eq.), anhydrous tetrahydrofuran (0.75 mL, 5V), and triethylamine (47.32 mg, 3.0 eq.) were added sequentially to the reaction flask. The mixture was slowly heated to 50 °C and stirred for 30 h. TLC (PE / EA = 1 / 1) analysis showed complete consumption of the starting materials. The reaction solution was directly purified by silica gel column chromatography (DCM / MeOH = 1 / 0-5 / 1). 48 mg of the target product, a white waxy substance, was obtained, with a yield of 28.96%.

[0207] The synthesized product underwent NMR analysis, and the characterization data are as follows:

[0208] 1H NMR(400MHz,Chloroform-d)δ4.41–4.22(m,2H),4.13–3.91(m,7H),3.74(t,J=4.2Hz ,2H),3.00–2.49(m,15H),1.62(t,J=7.1Hz,4H),1.26(s,66H),0.88(t,J=6.7Hz,6H).

[0209] The synthesized product was analyzed by high-resolution mass spectrometry, and the characterization data are as follows:

[0210] MS(m / z):[M+H] + =1063.82195.

[0211] Example 14 Synthesis of a novel phosphatidylcholine compound (14)

[0212] The structural formula of the novel phosphatidylcholine compound (14) is as follows:

[0213]

[0214] Step S4:

[0215]

[0216] Under nitrogen protection and at room temperature, M3 (180 mg crude product, 1.0 eq.), anhydrous tetrahydrofuran (0.9 mL, 5V), and DIPEA (60.43 mg, 3.0 eq.) were added sequentially to the reaction flask. The mixture was slowly heated to 50 °C and stirred for 30 h. TLC (PE / EA = 1 / 1) analysis showed complete consumption of the starting materials. The reaction solution was directly purified by silica gel column chromatography (DCM / MeOH = 1 / 0-5 / 1). 40 mg of the target product, a white waxy substance, was obtained, with a yield of 23.51%.

[0217] The synthesized product underwent NMR analysis, and the characterization data are as follows:

[0218] 1H NMR(400MHz,Chloroform-d)δ4.31(d,J=34.3Hz,2H),4.15–3.91(m,7H),3.75(t,J=4.3 Hz, 2H), 3.02–2.51 (m, 13H), 1.62 (t, J = 7.0Hz, 4H), 1.26 (s, 60H), 0.88 (t, J = 6.7Hz, 6H).

[0219] Example 15 Synthesis of a novel phosphatidylcholine compound (15)

[0220] The structural formula of the novel phosphatidylcholine compound (15) is as follows:

[0221]

[0222] Step S4:

[0223]

[0224] Under nitrogen protection and at room temperature, M3 (180 mg crude product, 1.0 eq.), anhydrous tetrahydrofuran (0.9 mL, 5V), and N,N-diethylmethylamine (48.91 mg, 3.0 eq.) were added sequentially to the reaction flask. The mixture was slowly heated to 50 °C and stirred for 30 h. TLC (PE / EA = 1 / 1) analysis showed complete consumption of the starting materials. The reaction solution was directly purified by silica gel column chromatography (DCM / MeOH = 1 / 0-5 / 1). 39.5 mg of the target product, a white waxy substance, was obtained, with a yield of 20.12%.

[0225] The synthesized product underwent NMR analysis, and the characterization data are as follows:

[0226] 1H NMR(400MHz,Chloroform-d)δ4.37–4.24(m,2H),4.06(t,J=6.7Hz,6H),3.95(t,J=6.7Hz,2H),3.7 4(t,J=4.1Hz,2H),2.98–2.41(m,18H),1.62(t,J=7.0Hz,4H),1.26(s,71H),0.88(t,J=6.8Hz,6H).

[0227] The synthesized product was analyzed by high-resolution mass spectrometry, and the characterization data are as follows:

[0228] MS(m / z):[M+H] + =1049.79375.

[0229] Example 16 Synthesis of a novel phosphatidylcholine compound (16)

[0230] The structural formula of the novel phosphatidylcholine compound (16) is as follows:

[0231]

[0232] Step S4:

[0233]

[0234] Under nitrogen protection and at room temperature, M3 (180 mg crude product, 1.0 eq.), anhydrous tetrahydrofuran (0.9 mL, 5V), and N-methyldipropylamine (64.65 mg, 3.0 eq.) were added sequentially to the reaction flask. The mixture was slowly heated to 50 °C and stirred for 30 h. TLC (PE / EA = 1 / 1) analysis showed complete consumption of the starting materials. The reaction solution was directly purified by silica gel column chromatography (DCM / MeOH = 1 / 0-5 / 1). 33.2 mg of the target product, a white waxy substance, was obtained, with a yield of 16.47%.

[0235] The synthesized product underwent NMR analysis, and the characterization data are as follows:

[0236] 1H NMR(400MHz,Chloroform-d)δ4.39–4.22(m,2H),4.06(t,J=6.8Hz,6H),3.95(d,J=7.3Hz,2H),3.7 4(t,J=4.1Hz,2H),2.98–2.47(m,17H),1.61(q,J=7.0Hz,4H),1.26(s,70H),0.88(t,J=6.7Hz,6H).

[0237] The synthesized product was analyzed by high-resolution mass spectrometry, and the characterization data are as follows:

[0238] MS(m / z):[M+H] + =1077.81804.

[0239] Example 17 Synthesis of a novel phosphatidylcholine compound (17)

[0240] The structural formula of the novel phosphatidylcholine compound (17) is as follows:

[0241]

[0242] Step S4:

[0243]

[0244] Under nitrogen protection and at room temperature, M3 (180 mg crude product, 1.0 eq.), anhydrous tetrahydrofuran (0.9 mL, 5V), and N,N-dimethylbutylamine (56.78 mg, 3.0 eq.) were added sequentially to the reaction flask. The mixture was slowly heated to 50 °C and stirred for 30 h. TLC (PE / EA = 1 / 1) analysis showed complete consumption of the starting materials. The reaction solution was directly purified by silica gel column chromatography (DCM / MeOH = 1 / 0-5 / 1). 29.1 mg of the target product, a white waxy substance, was obtained, with a yield of 14.63%.

[0245] The synthesized product underwent NMR analysis, and the characterization data are as follows:

[0246] 1H NMR(400MHz,Chloroform-d)δ4.36–4.23(m,2H),4.06(t,J=6.8Hz,6H),3.95(d,J=7.9Hz,2H),3.7 4(t,J=4.2Hz,2H),2.95–2.41(m,17H),1.62(p,J=6.9Hz,4H),1.26(s,68H),0.88(t,J=6.7Hz,6H).

[0247] The synthesized product was analyzed by high-resolution mass spectrometry, and the characterization data are as follows:

[0248] MS(m / z):[M+H] + =1063.81467.

[0249] Example 18 Synthesis of a novel phosphatidylcholine compound (18)

[0250] The structural formula of the novel phosphatidylcholine compound (18) is as follows:

[0251]

[0252] Step S4:

[0253]

[0254] Under nitrogen protection and at room temperature, M3 (180 mg crude product, 1.0 eq.), anhydrous tetrahydrofuran (0.9 mL, 5V), and N,N-dimethyl-n-octylamine (88.26 mg, 3.0 eq.) were added sequentially to the reaction flask. The mixture was slowly heated to 50 °C and stirred for 30 h. TLC (PE / EA = 1 / 1) analysis showed complete consumption of the starting materials. The reaction solution was directly purified by silica gel column chromatography (DCM / MeOH = 1 / 0-5 / 1). 35.40 mg of the target product, a white waxy substance, was obtained, with a yield of 16.90%.

[0255] The synthesized product underwent NMR analysis, and the characterization data are as follows:

[0256] 1H NMR(400MHz,Chloroform-d)δ4.45(dd,J=11.0,5.2Hz,1H),4.17–3.93(m,8H),3.76–3.61(m,2H),3 .24–2.47(m,17H),2.40–1.91(m,4H),1.56(p,J=6.9Hz,4H),1.31–1.06(m,68H),0.84–0.77(m,6H).

[0257] The synthesized product was analyzed by high-resolution mass spectrometry, and the characterization data are as follows:

[0258] MS(m / z):[M+H] + =1119.88601.

[0259] Example 19 Synthesis of a novel phosphatidylcholine compound (19)

[0260] The structural formula of the novel phosphatidylcholine compound (19) is as follows:

[0261]

[0262] Step S4:

[0263]

[0264] Under nitrogen protection and at room temperature, M3 (180 mg crude product, 1.0 eq.), anhydrous tetrahydrofuran (0.9 mL, 5V), and 1-methylpyrrolidine (47.78 mg, 3.0 eq.) were added sequentially to the reaction flask. The mixture was slowly heated to 50 °C and stirred for 30 h. TLC (PE / EA = 1 / 1) analysis showed complete consumption of the starting materials. The reaction solution was directly purified by silica gel column chromatography (DCM / MeOH = 1 / 0-5 / 1). 45.90 mg of the target product, a white waxy substance, was obtained, with a yield of 23.43%.

[0265] The synthesized product underwent NMR analysis, and the characterization data are as follows:

[0266] 1H NMR (400MHz, Chloroform-d) δ4.41–3.89(m,9H),3.78–3.61(m,2H),3.15–1.86(m,19H),1.54(q,J=7.1Hz,4H),1.31–0.97(m,66H),0.81(t,J=6.7Hz,6H).

[0267] The synthesized product was analyzed by high-resolution mass spectrometry, and the characterization data are as follows:

[0268] MS(m / z):[M+H] + =1147.79604.

[0269] Example 20 Synthesis of a novel phosphatidylcholine compound (20)

[0270] The structural formula of the novel phosphatidylcholine compound (20) is as follows:

[0271]

[0272] Step S4:

[0273]

[0274] Under nitrogen protection and at room temperature, M3 (180 mg crude product, 1.0 eq.), anhydrous tetrahydrofuran (0.9 mL, 5V), and N-methylpiperidine (55.65 mg, 3.0 eq.) were added sequentially to the reaction flask. The mixture was slowly heated to 50 °C and stirred for 30 h. TLC (PE / EA = 1 / 1) analysis showed complete consumption of the starting materials. The reaction solution was directly purified by silica gel column chromatography (DCM / MeOH = 1 / 0-5 / 1). 36.90 mg of the target product, a white waxy substance, was obtained, with a yield of 18.58%.

[0275] The synthesized product underwent NMR analysis, and the characterization data are as follows:

[0276] 1H NMR (400MHz, Chloroform-d) δ 4.42–3.88 (m, 9H), 3.73–3.61 (m, 2H), 3.10–2.49 (m, 15H), 1.55 (t, J = 7.1Hz, 4H), 1.19 (s, 66H), 0.81 (t, J = 6.8Hz, 6H).

[0277] The synthesized product was analyzed by high-resolution mass spectrometry, and the characterization data are as follows:

[0278] MS(m / z):[M+H] + =1061.79010.

[0279] Example 21 Synthesis of a novel phosphatidylcholine compound (21)

[0280] The structural formula of the novel phosphatidylcholine compound (21) is as follows:

[0281]

[0282] Step S4:

[0283]

[0284] Under nitrogen protection and at room temperature, M3 (180 mg crude product, 1.0 eq.), anhydrous tetrahydrofuran (0.9 mL, 5V), and N-ethylpiperidine (63.52 mg, 3.0 eq.) were added sequentially to the reaction flask. The mixture was slowly heated to 50 °C and stirred for 30 h. TLC (PE / EA = 1 / 1) analysis showed complete consumption of the starting materials. The reaction solution was directly purified by silica gel column chromatography (DCM / MeOH = 1 / 0-5 / 1). 37.60 mg of the target product, a white waxy substance, was obtained, with a yield of 18.69%.

[0285] The synthesized product underwent NMR analysis, and the characterization data are as follows:

[0286] 1H NMR(400MHz,Chloroform-d)δ4.46–4.35(m,1H),4.15–3.91(m,8H),3.75–3.62(m,2H),3.20–2 .62(m,16H),1.94(d,J=6.4Hz,3H),1.56(t,J=7.1Hz,4H),1.19(s,65H),0.81(t,J=6.7Hz,6H).

[0287] Example 22 Synthesis of a novel phosphatidylcholine compound (22)

[0288] The structural formula of the novel phosphatidylcholine compound (22) is as follows:

[0289]

[0290] Step S4:

[0291]

[0292] Under nitrogen protection and at room temperature, M3 (180 mg crude product, 1.0 eq.), anhydrous tetrahydrofuran (0.9 mL, 5V), and N,N-dimethylcyclohexylamine (71.39 mg, 3.0 eq.) were added sequentially to the reaction flask. The mixture was slowly heated to 50 °C and stirred for 30 h. TLC (PE / EA = 1 / 1) analysis showed complete consumption of the starting materials. The reaction solution was directly purified by silica gel column chromatography (DCM / MeOH = 1 / 0-5 / 1). 40.80 mg of the target product, a white waxy substance, was obtained, with a yield of 20.02%.

[0293] The synthesized product underwent NMR analysis, and the characterization data are as follows:

[0294] 1H NMR(400MHz,Chloroform-d)δ4.36(d,J=12.2Hz,1H),4.20–3.87(m,8H),3.75–3.61(m,2H), 3.42(s,1H),3.09–2.35(m,17H),1.55(t,J=7.1Hz,4H),1.19(s,65H),0.81(t,J=6.7Hz,6H).

[0295] The synthesized product was analyzed by high-resolution mass spectrometry, and the characterization data are as follows:

[0296] MS(m / z):[M+H] + =1089.83038.

[0297] Example 23 Synthesis of a novel phosphatidylcholine compound (23)

[0298] The structural formula of the novel phosphatidylcholine compound (23) is as follows:

[0299]

[0300] Step S3:

[0301]

[0302] M2 (150 mg, 1.0 eq.) and anhydrous DCM (1.5 mL, 10V) were added sequentially to the reaction flask. The mixture was cooled to 4°C in an ice bath, and 2-chloro-2-oxo-1,3,2-dioxophosphazenecyclopentane (51.35 mg, 2.0 eq.) was added. After the addition was complete, the mixture was stirred for 5 minutes. Trimethylamine (2.0 min THF) (0.25 mL, 2.5 eq.) was then slowly added dropwise. One hour after the addition was complete, TLC (PE / EA = 1 / 1) showed that the starting material had been completely consumed, and the reaction was considered complete. The reaction solution was used directly in the next step without further purification. The estimated weight of the crude product was 160 mg.

[0303] Step S4:

[0304]

[0305] Under nitrogen protection and at room temperature, trimethylamine (2M in THF) (0.15 mL, 1.5 eq.) was added to crude product solution M3 (160 mg crude product, 1.0 eq.) in the reaction flask with stirring. The mixture was slowly heated to 50 °C and stirred for 30 h. TLC (PE / EA = 1 / 1) analysis showed complete consumption of the starting material. The reaction solution was directly purified by silica gel column chromatography (DCM / MeOH = 1 / 0-5 / 1). 44.7 mg of the target product was obtained as a white solid, with a yield of 26.28%.

[0306] The synthesized product underwent NMR analysis, and the characterization data are as follows:

[0307] 1H NMR(400MHz,Chloroform-d)δ4.30(td,J=11.1,10.4,5.7Hz,2H),4.05(q,J=5.4Hz,2H),3.94(s,2H),3.74( d,J=5.4Hz,2H),2.91–2.78(m,9H),2.74–2.41(m,10H),1.66–1.50(m,4H),1.26(s,60H),0.91–0.83(m,6H).

[0308] Example 24 Synthesis of a novel phosphatidylcholine compound (24)

[0309] The structural formula of the novel phosphatidylcholine compound (24) is as follows:

[0310]

[0311] Step S3:

[0312]

[0313] M2 (150 mg, 1.0 eq.) and anhydrous DCM (1.5 mL, 10V) were added sequentially to the reaction flask. The mixture was cooled to 4°C in an ice bath, and 2-chloro-2-oxo-1,3,2-dioxophosphazenecyclopentane (51.35 mg, 2.0 eq.) was added. After the addition was complete, the mixture was stirred for 5 minutes. Triethylamine (45.59 mg, 2.5 eq.) was then slowly added dropwise. One hour after the addition was complete, TLC (PE / EA = 1 / 1) showed that the starting material had been completely consumed, and the reaction was considered complete. The reaction solution was used directly in the next step without further purification. The estimated weight of the crude product was 160 mg.

[0314] Step S4:

[0315]

[0316] Under nitrogen protection and at room temperature, triethylamine (25.88 mg, 1.5 eq.) was added to crude product solution M3 (160 mg crude product, 1.0 eq.) in the reaction flask with stirring. The mixture was slowly heated to 50 °C and stirred for 30 h. TLC (PE / EA = 1 / 1) analysis showed complete consumption of the starting material. The reaction solution was directly purified by silica gel column chromatography (DCM / MeOH = 1 / 0-5 / 1). 25 mg of the target product, a white solid, was obtained, with a yield of 14.10%.

[0317] The synthesized product underwent NMR analysis, and the characterization data are as follows:

[0318] 1H NMR(400MHz,Chloroform-d)δ4.36–4.21(m,2H),4.05(q,J=5.6Hz,2H),3.95(s,2H),3.79–3.70(m,2H),2 .91–2.77(m,9H),2.74–2.46(m,9H),1.56(dq,J=15.0,7.6,6.7Hz,4H),1.26(s,60H),0.93–0.84(m,6H).

[0319] Example 25 Synthesis of a novel phosphatidylcholine compound (25)

[0320] The structural formula of the novel phosphatidylcholine compound (25) is as follows:

[0321]

[0322] Step S3:

[0323]

[0324] M2 (150 mg, 1.0 eq.) and anhydrous DCM (1.5 mL, 10V) were added sequentially to the reaction flask. The mixture was cooled to 4°C in an ice bath, and 2-chloro-2-oxo-1,3,2-dioxophosphazenecyclopentane (51.35 mg, 2.0 eq.) was added. After the addition was complete, the mixture was stirred for 5 minutes. DIPEA (58.23 mg, 2.5 eq.) was then slowly added dropwise. One hour after the addition was complete, TLC (PE / EA = 1 / 1) showed that the starting material had been completely consumed, and the reaction was considered complete. The reaction solution was used directly in the next step without further purification. The estimated weight of the crude product was 160 mg.

[0325] Step S4:

[0326]

[0327] Under nitrogen protection and at room temperature, crude product solution M3 (160 mg crude product, 1.0 eq.) was added to the reaction flask with stirring, followed by the addition of DIPEA (33.05 mg, 1.5 eq.). The mixture was slowly heated to 50 °C and stirred for 30 h. TLC (PE / EA = 1 / 1) analysis showed complete consumption of the reactants. The reaction solution was then directly purified by silica gel column chromatography (DCM / MeOH = 1 / 0-5 / 1). 30 mg of the target product, a white solid, was obtained, with a yield of 16.48%.

[0328] The synthesized product underwent NMR analysis, and the characterization data are as follows:

[0329] 1H NMR (400MHz, Chloroform-d) δ4.28 (d, J=4.7Hz, 2H), 4.14–3.89 (m, 4H), 3.72 (d, J=7. 1Hz, 2H), 2.93–2.46 (m, 22H), 1.62–1.49 (m, 4H), 1.26 (s, 52H), 0.88 (t, J = 6.7Hz, 6H).

[0330] Example 26 Synthesis of a novel phosphatidylcholine compound (26)

[0331] The structural formula of the novel phosphatidylcholine compound (26) is as follows:

[0332]

[0333] Step S4:

[0334]

[0335] Under nitrogen protection and at room temperature, M3 (160 mg crude product, 1.0 eq.), anhydrous tetrahydrofuran (0.8 mL, 5V), and N,N-diethylmethylamine (44.59 mg, 3.0 eq.) were added sequentially to the reaction flask. The mixture was slowly heated to 50 °C and stirred for 30 h. TLC (PE / EA = 1 / 1) analysis showed complete consumption of the starting materials. The reaction solution was directly purified by silica gel column chromatography (DCM / MeOH = 1 / 0-5 / 1). 64 mg of the target product, a white waxy substance, was obtained, with a yield of 36.60%.

[0336] The synthesized product underwent NMR analysis, and the characterization data are as follows:

[0337] 1H NMR(400MHz,Chloroform-d)δ4.30(d,J=5.9Hz,1H),4.01(dd,J=46.5,6.9Hz,4H),3.79– 3.65(m,3H),2.96–2.47(m,20H),1.65–1.50(m,4H),1.26(s,62H),0.88(t,J=6.7Hz,6H).

[0338] The synthesized product was analyzed by high-resolution mass spectrometry, and the characterization data are as follows:

[0339] MS(m / z):[M+H] + =1025.68314.

[0340] Example 27 Synthesis of a novel phosphatidylcholine compound (27)

[0341] The structural formula of the novel phosphatidylcholine compound (27) is as follows:

[0342]

[0343] Step S4:

[0344]

[0345] Under nitrogen protection and at room temperature, M3 (160 mg crude product, 1.0 eq.), anhydrous tetrahydrofuran (0.8 mL, 5V), and N-methyldipropylamine (58.94 mg, 3.0 eq.) were added sequentially to the reaction flask. The mixture was slowly heated to 50 °C and stirred for 30 h. TLC (PE / EA = 1 / 1) analysis showed complete consumption of the starting materials. The reaction solution was directly purified by silica gel column chromatography (DCM / MeOH = 1 / 0-5 / 1). 30.60 mg of the target product, a white waxy substance, was obtained, with a yield of 17.03%.

[0346] The synthesized product underwent NMR analysis, and the characterization data are as follows:

[0347] 1H NMR(400MHz,Chloroform-d)δ4.34–4.26(m,2H),4.13–4.02(m,2H),3.96(d,J=7.6Hz,2H),3.73(dd,J=6.0,3 .0Hz,2H),3.00–2.55(m,19H),2.01(d,J=6.4Hz,2H),1.64–1.50(m,4H),1.26(s,63H),0.88(t,J=6.7Hz,6H).

[0348] Example 28 Synthesis of a novel phosphatidylcholine compound (28)

[0349] The structural formula of the novel phosphatidylcholine compound (28) is as follows:

[0350]

[0351] Step S4:

[0352]

[0353] Under nitrogen protection and at room temperature, M3 (160 mg crude product, 1.0 eq.), anhydrous tetrahydrofuran (0.8 mL, 5V), and N,N-dimethylbutylamine (51.76 mg, 3.0 eq.) were added sequentially to the reaction flask. The mixture was slowly heated to 50 °C and stirred for 30 h. TLC (PE / EA = 1 / 1) analysis showed complete consumption of the starting materials. The reaction solution was directly purified by silica gel column chromatography (DCM / MeOH = 1 / 0-5 / 1). 31.80 mg of the target product, a white waxy substance, was obtained, with a yield of 17.94%.

[0354] The synthesized product underwent NMR analysis, and the characterization data are as follows:

[0355] 1H NMR(400MHz,Chloroform-d)δ4.36–4.26(m,2H),4.06(q,J=5.7,5.2Hz,2H),3.99–3.90(m,2H),3.73(dd,J=5.8,2.9 Hz, 2H), 2.97–2.55 (m, 20H), 2.01 (d, J = 6.2Hz, 3H), 1.56 (p, J = 7.6, 6.9Hz, 4H), 1.26 (s, 62H), 0.88 (t, J = 6.8Hz, 6H).

[0356] Example 29 Synthesis of a novel phosphatidylcholine compound (29)

[0357] The structural formula of the novel phosphatidylcholine compound (29) is as follows:

[0358]

[0359] Step S3:

[0360]

[0361] M2 (150 mg, 1.0 eq.) and anhydrous tetrahydrofuran (1.5 mL, 10V) were added sequentially to the reaction flask. The mixture was cooled to 4°C in an ice bath, and 2-chloro-2-oxo-1,3,2-dioxophosphazenecyclopentane (51.35 mg, 2.0 eq.) was added. After the addition was complete, the mixture was stirred for 5 minutes. Then, N,N-dimethyl-n-octylamine (70.86 mg, 2.5 eq.) was slowly added dropwise. One hour after the addition was complete, TLC (PE / EA = 1 / 1) showed that the starting material had been completely consumed, and the reaction was considered complete. The reaction solution was used directly in the next step without further purification. The estimated weight of the crude product was 160 mg.

[0362] Step S4:

[0363]

[0364] Under nitrogen protection and at room temperature, N,N-dimethyl-n-octylamine (40.23 mg, 1.5 eq.) was added to crude product solution M3 (160 mg crude product, 1.0 eq.) in the reaction flask with stirring. The mixture was slowly heated to 50 °C and stirred for 30 h. TLC (PE / EA = 1 / 1) analysis showed complete consumption of the starting material. The reaction solution was directly purified by silica gel column chromatography (DCM / MeOH = 1 / 0-5 / 1). 39.20 mg of the target product (white solid) was obtained, with a yield of 20.98%.

[0365] The synthesized product underwent NMR analysis, and the characterization data are as follows:

[0366] 1H NMR(400MHz,Chloroform-d)δ4.31(t,J=4.9Hz,2H),4.13–4.04(m,2H),3.97(d,J=7.4Hz,2H),3.78–3.69(m,2H) ,2.88(dp,J=14.9,7.5,6.9Hz,10H),2.79–2.56(m,11H),1.60–1.51(m,4H),1.26(s,63H),0.88(t,J=6.8Hz,6H).

[0367] Example 30 Synthesis of a novel phosphatidylcholine compound (30)

[0368] The structural formula of the novel phosphatidylcholine compound (30) is as follows:

[0369]

[0370] Step S3:

[0371]

[0372] M2 (150 mg, 1.0 eq.) and anhydrous tetrahydrofuran (1.5 mL, 10V) were added sequentially to the reaction flask. The mixture was cooled to 4°C in an ice bath, and 2-chloro-2-oxo-1,3,2-dioxophosphazenecyclopentane (51.35 mg, 2.0 eq.) was added. After the addition was complete, the mixture was stirred for 5 minutes. Then, 1-methylpyrrolidine (38.36 mg, 2.5 eq.) was slowly added dropwise. One hour after the addition was complete, TLC (PE / EA = 1 / 1) showed that the starting material had been completely consumed, and the reaction was considered complete. The reaction solution was used directly in the next step without further purification. The estimated weight of the crude product was 160 mg.

[0373] Step S4:

[0374]

[0375] Under nitrogen protection and at room temperature, crude product solution M3 (160 mg crude product, 1.0 eq.) was added to the reaction flask with stirring, followed by the addition of 1-methylpyrrolidine (21.78 mg, 1.5 eq.). The mixture was slowly heated to 50 °C and stirred for 30 h. TLC (PE / EA = 1 / 1) analysis showed complete consumption of the starting material. The reaction solution was then directly purified by silica gel column chromatography (DCM / MeOH = 1 / 0-5 / 1). 42 mg of the target product, a white solid, was obtained, with a yield of 24.07%.

[0376] The synthesized product underwent NMR analysis, and the characterization data are as follows:

[0377] 1H NMR(400MHz,Chloroform-d)δ4.31(t,J=5.1Hz,2H),4.07(q,J=5.8,5.2Hz,2H),3.97(d,J=7.7Hz,2H),3.83–3.64(m ,3H),2.88(dp,J=14.9,7.4,6.9Hz,9H),2.79–2.57(m,9H),1.65–1.49(m,4H),1.26(s,60H),0.88(t,J=6.7Hz,6H).

[0378] The synthesized product was analyzed by high-resolution mass spectrometry, and the characterization data are as follows:

[0379] MS(m / z):[M+H] + =1023.61454.

[0380] Example 31 Synthesis of a novel phosphatidylcholine compound (31)

[0381] The structural formula of the novel phosphatidylcholine compound (31) is as follows:

[0382]

[0383] Step S3:

[0384]

[0385] M2 (150 mg, 1.0 eq.) and anhydrous tetrahydrofuran (1.5 mL, 10V) were added sequentially to the reaction flask. The mixture was cooled to 4°C in an ice bath, and 2-chloro-2-oxo-1,3,2-dioxophosphazenecyclopentane (51.35 mg, 2.0 eq.) was added. After the addition was complete, the mixture was stirred for 5 minutes. N-methylpiperidine (44.68 mg, 2.5 eq.) was then slowly added dropwise. One hour after the addition was complete, TLC (PE / EA = 1 / 1) showed that the starting material had been completely consumed, and the reaction was considered complete. The reaction solution was used directly in the next step without further purification. The estimated weight of the crude product was 160 mg.

[0386] Step S4:

[0387]

[0388] Under nitrogen protection and at room temperature, N-methylpiperidine (25.36 mg, 1.5 eq.) was added to crude product solution M3 (160 mg crude product, 1.0 eq.) in the reaction flask with stirring. The mixture was slowly heated to 50 °C and stirred for 30 h. TLC (PE / EA = 1 / 1) analysis showed complete consumption of the starting material. The reaction solution was directly purified by silica gel column chromatography (DCM / MeOH = 1 / 0-5 / 1). 50.30 mg of the target product (white solid) was obtained, with a yield of 28.43%.

[0389] The synthesized product underwent NMR analysis, and the characterization data are as follows:

[0390] 1H NMR(400MHz,Chloroform-d)δ4.35–4.26(m,2H),4.06(q,J=5.6Hz,2H),3.95(q,J=6.9,5.0Hz,2H),3.73(t,J=4. 3Hz, 2H), 2.87 (q, J = 7.4, 7.0Hz, 9H), 2.77–2.57 (m, 10H), 1.63–1.49 (m, 4H), 1.26 (s, 61H), 0.88 (t, J = 6.8Hz, 6H).

[0391] The synthesized product was analyzed by high-resolution mass spectrometry, and the characterization data are as follows:

[0392] MS(m / z):[M+H] + =1037.68125.

[0393] Example 32 Synthesis of a novel phosphatidylcholine compound (32)

[0394] The structural formula of the novel phosphatidylcholine compound (32) is as follows:

[0395]

[0396] Step S3:

[0397]

[0398] M2 (150 mg, 1.0 eq.) and anhydrous tetrahydrofuran (1.5 mL, 10V) were added sequentially to the reaction flask. The mixture was cooled to 4°C in an ice bath, and 2-chloro-2-oxo-1,3,2-dioxophosphazenecyclopentane (51.35 mg, 2.0 eq.) was added. After the addition was complete, the mixture was stirred for 5 minutes. N-ethylpiperidine (51.00 mg, 2.5 eq.) was then slowly added dropwise. One hour after the addition was complete, TLC (PE / EA = 1 / 1) showed that the starting material had been completely consumed, and the reaction was considered complete. The reaction solution was used directly in the next step without further purification. The estimated weight of the crude product was 160 mg.

[0399] Step S4:

[0400]

[0401] Under nitrogen protection and at room temperature, N-ethylpiperidine (28.95 mg, 1.5 eq.) was added to crude product solution M3 (160 mg crude product, 1.0 eq.) in the reaction flask with stirring. The mixture was slowly heated to 50 °C and stirred for 30 h. TLC (PE / EA = 1 / 1) analysis showed complete consumption of the starting material. The reaction solution was directly purified by silica gel column chromatography (DCM / MeOH = 1 / 0-5 / 1). 64 mg of the target product, a white solid, was obtained, with a yield of 35.69%.

[0402] The synthesized product underwent NMR analysis, and the characterization data are as follows:

[0403] 1H NMR(400MHz,Chloroform-d)δ4.37–4.19(m,2H),4.13–4.02(m,2H),3.96(d,J=7.3Hz,2H),3.74(d,J=4. 9Hz, 2H), 2.93–2.81 (m, 8H), 2.76–2.57 (m, 9H), 1.65–1.49 (m, 4H), 1.26 (s, 61H), 0.88 (t, J = 6.8Hz, 6H).

[0404] Example 33 Synthesis of a novel phosphatidylcholine compound (33)

[0405] The structural formula of the novel phosphatidylcholine compound (33) is as follows:

[0406]

[0407] Step S3:

[0408]

[0409] M2 (150 mg, 1.0 eq.) and anhydrous tetrahydrofuran (1.5 mL, 10V) were added sequentially to the reaction flask. The mixture was cooled to 4°C in an ice bath, and 2-chloro-2-oxo-1,3,2-dioxophosphazenecyclopentane (51.35 mg, 2.0 eq.) was added. After the addition was complete, the mixture was stirred for 5 minutes. Then, N,N-dimethylcyclohexylamine (57.32 mg, 2.5 eq.) was slowly added dropwise. One hour after the addition was complete, TLC (PE / EA = 1 / 1) showed that the starting material had been completely consumed, and the reaction was considered complete. The reaction solution was used directly in the next step without further purification. The estimated weight of the crude product was 160 mg.

[0410] Step S4:

[0411]

[0412] Under nitrogen protection and at room temperature, N,N-dimethylcyclohexylamine (32.54 mg, 1.5 eq.) was added to crude product solution M3 (160 mg crude product, 1.0 eq.) in the reaction flask with stirring. The mixture was slowly heated to 50 °C and stirred for 30 h. TLC (PE / EA = 1 / 1) analysis showed complete consumption of the starting material. The reaction solution was directly purified by silica gel column chromatography (DCM / MeOH = 1 / 0-5 / 1). 71 mg of the target product, a white solid, was obtained, with a yield of 39.08%.

[0413] The synthesized product underwent NMR analysis, and the characterization data are as follows:

[0414] 1H NMR(400MHz,Chloroform-d)δ4.30(d,J=6.3Hz,2H),4.06(dd,J=7.0,4.6Hz,2H),3.94(q,J=7.0,5.1Hz,2H),3.73 (t,J=4.1Hz,2H),2.86(t,J=7.4Hz,9H),2.74–2.55(m,10H),1.63–1.49(m,4H),1.26(s,65H),0.90–0.85(m,6H).

[0415] The synthesized product was analyzed by high-resolution mass spectrometry, and the characterization data are as follows:

[0416] MS(m / z):[M+H] + =1065.72139.

[0417] Example 34 Synthesis of a novel phosphatidylcholine compound (34)

[0418] The structural formula of the novel phosphatidylcholine compound (34) is as follows:

[0419]

[0420] Step S2:

[0421]

[0422] At room temperature, M1 (1.5 g, 1.0 eq.), SM3 (containing 0.05% BHT stabilizer) (3.83 g, 2.2 eq.), and acetic acid (21.73 mg, 0.05 eq.) were added sequentially to a reaction flask. The mixture was stirred and heated to 70°C for 40 h. TLC (DCM / MeOH = 20 / 1) analysis showed that the starting materials were almost completely consumed. The reaction solution was directly purified by silica gel column chromatography (DCM / MeOH = 1 / 0-100 / 1). 2.4 g of the target product, a pale yellow oil, was obtained, with a yield of 48.20%.

[0423] The NMR analysis of the synthesized intermediate compound M2 yielded the following characterization data:

[0424] 1H NMR(400MHz,Chloroform-d)δ4.36–4.27(m,1H),4.24–4.08(m,2H),3.97(d,J=5.8Hz,4H),3.83(t,J=4.6Hz,2H),2.87–2.69(m ,6H),2.69–2.56(m,6H),2.45(t,J=7.2Hz,4H),1.78–1.54(m,3H),1.27(tt,J=7.3,4.4Hz,39H),0.89(td,J=6.6,4.2Hz,12H).

[0425] Step S3:

[0426]

[0427] M2 (150 mg, 1.0 eq.) and anhydrous tetrahydrofuran (1.5 mL, 10V) were added sequentially to the reaction flask. The mixture was cooled to 4°C in an ice bath, and 2-chloro-2-oxo-1,3,2-dioxophosphazenecyclopentane (62.12 mg, 2.0 eq.) was added. After the addition was complete, the mixture was stirred for 5 minutes. N,N-diethylmethylamine (47.51 mg, 2.5 eq.) was then slowly added dropwise. One hour after the addition was complete, TLC (PE / EA = 1 / 1) showed that the starting material had been completely consumed, and the reaction was considered complete. The reaction solution was used directly in the next step without further purification. The estimated weight of the crude product was 160 mg.

[0428] Step S4:

[0429]

[0430] Under nitrogen protection and at room temperature, N,N-diethylmethylamine (26.34 mg, 1.5 eq.) was added to crude product solution M3 (160 mg crude product, 1.0 eq.) in the reaction flask with stirring. The mixture was slowly heated to 50 °C and stirred for 30 h. TLC (PE / EA = 1 / 1) analysis showed complete consumption of the starting material. The reaction solution was directly purified by silica gel column chromatography (DCM / MeOH = 1 / 0-5 / 1). 65.70 mg of the target product, a colorless oil, was obtained, with a yield of 25.74%.

[0431] The synthesized product underwent NMR analysis, and the characterization data are as follows:

[0432] 1H NMR(400MHz,Chloroform-d)δ4.32(ddd,J=38.0,12.4,7.0Hz,2H),4.14–4.05(m,4H),3.98(d,J=5.8Hz,4H),3.70(t ,J=5.8Hz,2H),3.02–2.44(m,16H),1.61(q,J=5.7Hz,2H),1.28(hept,J=7.1Hz,43H),0.89(td,J=6.6,4.2Hz,12H).

[0433] Example 35 Synthesis of a novel phosphatidylcholine compound (35)

[0434] The structural formula of the novel phosphatidylcholine compound (35) is as follows:

[0435]

[0436] Step S3:

[0437]

[0438] M2 (150 mg, 1.0 eq.) and anhydrous tetrahydrofuran (1.5 mL, 10V) were added sequentially to the reaction flask. The mixture was cooled to 4°C in an ice bath, and 2-chloro-2-oxo-1,3,2-dioxophosphazenecyclopentane (62.12 mg, 2.0 eq.) was added. After the addition was complete, the mixture was stirred for 5 minutes. N-methyldipropylamine (62.80 mg, 2.5 eq.) was then slowly added dropwise. One hour after the addition was complete, TLC (PE / EA = 1 / 1) showed that the starting material had been completely consumed, and the reaction was considered complete. The reaction solution was used directly in the next step without further purification. The estimated weight of the crude product was 160 mg.

[0439] Step S4:

[0440]

[0441] Under nitrogen protection and at room temperature, N-methyldipropylamine (34.82 mg, 1.5 eq.) was added to crude product solution M3 (160 mg crude product, 1.0 eq.) in the reaction flask with stirring. The mixture was slowly heated to 50 °C and stirred for 30 h. TLC (PE / EA = 1 / 1) analysis showed complete consumption of the starting material. The reaction solution was directly purified by silica gel column chromatography (DCM / MeOH = 1 / 0-5 / 1). 137 mg of the target product, a colorless oil, was obtained, with a yield of 52.02%.

[0442] The synthesized product underwent NMR analysis, and the characterization data are as follows:

[0443] 1H NMR(400MHz,Chloroform-d)δ4.42–4.23(m,2H),4.13–4.02(m,4H),3.98(d,J=5.8Hz,4H),3.70(t,J=5 .7Hz,3H),2.99–2.41(m,16H),1.61(q,J=5.6Hz,2H),1.39–1.00(m,39H),0.89(td,J=6.6,4.2Hz,12H).

[0444] Example 36 Synthesis of a novel phosphatidylcholine compound (36)

[0445] The structural formula of the novel phosphatidylcholine compound (36) is as follows:

[0446]

[0447] Step S3:

[0448]

[0449] M2 (150 mg, 1.0 eq.) and anhydrous tetrahydrofuran (1.5 mL, 10V) were added sequentially to the reaction flask. The mixture was cooled to 4°C in an ice bath, and 2-chloro-2-oxo-1,3,2-dioxophosphazenecyclopentane (62.12 mg, 2.0 eq.) was added. After the addition was complete, the mixture was stirred for 5 minutes. N,N-dimethylbutylamine (55.15 mg, 2.5 eq.) was then slowly added dropwise. One hour after the addition was complete, TLC (PE / EA = 1 / 1) showed that the starting material had been completely consumed, and the reaction was considered complete. The reaction solution was used directly in the next step without further purification. The estimated weight of the crude product was 160 mg.

[0450] Step S4:

[0451]

[0452] Under nitrogen protection and at room temperature, N,N-dimethylbutylamine (30.08 mg, 1.5 eq.) was added to crude product solution M3 (160 mg crude product, 1.0 eq.) in the reaction flask with stirring. The mixture was slowly heated to 50 °C and stirred for 30 h. TLC (PE / EA = 1 / 1) analysis showed complete consumption of the starting material. The reaction solution was directly purified by silica gel column chromatography (DCM / MeOH = 1 / 0-5 / 1). 100 mg of the target product, a colorless oil, was obtained, with a yield of 55.44%.

[0453] The synthesized product underwent NMR analysis, and the characterization data are as follows:

[0454] 1H NMR(400MHz,Chloroform-d)δ4.40–4.24(m,2H),4.14–4.04(m,4H),3.98(d,J=5.8Hz,4H),3.70(t,J=5.7 Hz,2H),3.00–2.46(m,15H),1.62(t,J=6.0Hz,2H),1.37–1.12(m,37H),0.89(td,J=7.0,6.6,4.2Hz,12H).

[0455] LNP preparation and screening

[0456] mRNA: All mRNAs used in these studies were synthesized and purified by Starray Pharma (Suzhou) Co., Ltd. in vitro. Fluci mRNA encodes luciferase. After transfection into cells or administration to animals, it can express luciferase protein. When luciferin is added, it can emit light. The intensity of its light emission is positively correlated with the expression of luciferase. It is used to evaluate the delivery capability of the delivery vector.

[0457] Formula 1LNP: Formula 1LNP contains four lipids, namely SM-102, the lipid of this invention, cholesterol, and DMG-PEG2000; the molar percentages of the four lipids are 46% SM-102, 8% lipid of this invention, 43.95% cholesterol, and 2.05% DMG-PEG2000; the mass ratio of total lipids to Fluci mRNA is 18:1.

[0458] SM-102LNP: SM-102LNP contains four lipids, with the following molar percentages: 50% SM-102, 10% DSPC, 38.5% cholesterol, and 1.5% DMG-PEG2000; the total lipid to Fluci mRNA mass ratio is 19:1.

[0459] LNP preparation: Four lipids were dissolved in ethanol according to their corresponding formulation ratios, and Fluci mRNA was dissolved in 10 mM sodium citrate solution (pH = 4.0). The crude LNP product was prepared by mixing the ethanol phase (four lipids) and the aqueous phase (mRNA) at a volume ratio of 1:3. The crude LNP product was diluted 3 times with PBS, purified using a 100 kD ultrafiltration tube, and the ethanol was replaced with PBS. The product was then filtered through a 0.22 μm sterile filter to obtain LNPs encapsulated with Fluci mRNA.

[0460] Physicochemical parameters of LNP were determined: particle size, polydispersity index and zeta potential were determined using Zetasizer Pro (Malvern Panalytical); encapsulation efficiency of LNP was determined using Quant-it RiboGreen assay (Thermo Fisher Scientific).

[0461] The lipid structure of SM-102 is as follows:

[0462]

[0463] Example 37: In vivo delivery and expression activity of the novel lipid formulation 1LNP

[0464] Formula 1 LNP was prepared using Fluci mRNA as the nucleic acid component; Fluci is luciferase mRNA, a common reporter gene expressed in cells, which uses luciferin as a substrate to catalyze the emission of biofluorescence, and the abundance of luciferase expression is assessed by collecting the fluorescence intensity; the components and physicochemical properties of the LNP formulation are shown in Table 1:

[0465] Table 1 represents the physicochemical properties of LNP formulations.

[0466]

[0467]

[0468] Example 38: In vitro delivery and expression activity of the novel lipid formulation 1LNP

[0469] Following Example 37, LNPs prepared using Fluci mRNA as a nucleic acid component were transfected into in vitro cultured HEK293T and Huh7 cell lines. Cell viability and Fluci expression levels after transfection were measured using the commercially available ONE GLO Kit, and Fluci expression levels were normalized using cell viability. Both cell lines were plated in 96-well plates at approximately 2E4 cells / well, and 25 ng of LNPs were transfected into each well.

[0470] Figure 2 Figure 3 The formulations of compounds (1) to (22) of the 1LNP formulation showed high in vitro delivery expression activity in HEK293T and Huh7 cells.

[0471] Example 39 Comparison of targeted delivery expression activities of compound (1) and compound (4) formulations 1LNP and SM-102LNP

[0472] In accordance with Example 37, the lipid formulations of compounds (1) and (4) 1LNP and SM-102LNP were prepared using FlucimRNA as the nucleic acid component; the abundance of luciferase expression was assessed by collecting fluorescence intensity.

[0473] C57 mice (SPF grade, female, 6-8 weeks old) were administered the drug once via tail vein on day 0, at a dose of 0.25 mpk and a volume of 200 μL. Six hours after administration, the mice were injected intraperitoneally with 30 mg of luciferase substrate. Organs were dissected within 10-15 minutes, and imaging was performed using the PerkinElmer IVIS small animal imaging system. Fluci expression levels were assessed by fluorescence intensity.

[0474] Figure 4 The lipid formulations of compounds (1) and (4) showed that Fluci in the 1LNP formulation had good spleen-targeting specificity, while Fluci in the SM-102LNP formulation had good liver-targeting specificity and high expression efficiency. Among them, the expression level of Fluci in the 1LNP formulation of compounds (1) and (4) was the highest, reaching over 90%, while the expression level of Fluci in the liver of SM-102LNP was the highest, reaching over 80%. Figure 5 As shown.

[0475] Example 40: Formulation of novel lipids (2), (13), (24), (5), (16) and (27) and their delivery and expression activities.

[0476] The formulation of novel lipids of compounds (2), (13), (24), (5), (16) and (27) was carried out using Fluci mRNA as a nucleic acid component; the abundance of luciferase expression was assessed by collecting fluorescence intensity.

[0477] The components and physicochemical properties of the LNP formulation are shown in Table 2.

[0478] Table 2 Physicochemical properties of LNP formulations

[0479]

[0480]

[0481] C57 mice (SPF grade, female, 6-8 weeks old) were administered the drug once via tail vein on day 0, at a dose of 0.25 mpk and a volume of 200 μL. Six hours after administration, the mice were injected intraperitoneally with 30 mg of luciferase substrate. Organs were dissected within 10-15 minutes, and imaging was performed using the PerkinElmer IVIS small animal imaging system. Fluci expression levels were assessed by fluorescence intensity.

[0482] Figure 6 The novel lipid formulations of compounds (2), (13), (24), (5), (16), and (27) showed significant Fluci expression in both the liver and spleen, with higher expression efficiency in the spleen. Compound (2) showed the highest Fluci expression level in the spleen, exceeding 80%. Figure 7 As shown.

[0483] The foregoing has shown and described the basic principles, main features, and advantages of the present invention. Those skilled in the art should understand that the present invention is not limited to the above embodiments. The embodiments and descriptions in the specification are merely illustrative of the principles of the invention. Various changes and modifications can be made to the invention without departing from its spirit and scope, and all such changes and modifications fall within the scope of the present invention as claimed. The scope of protection of this invention is defined by the appended claims and their equivalents.

Claims

1. A phosphatidylcholine compound represented by formula (I) or formula (II), a pharmaceutically acceptable salt, tautomer or stereoisomer of the compound, ###0001### (I) (II) wherein X is selected from -O- or -S-; R1 and R2 are selected from -H or a group of the following structure, and R1 and R2 are not simultaneously -H, ###0002### and the compound of formula (I) or formula (II) has a structure of formula (la) or (lla): ###0003### (la) (lla) and the compound of formula (I) has a structure as shown below: ###0004### ； 6. Use of a phosphatidylcholine compound represented by the structure of formula (la) according to claim 4, a pharmaceutically acceptable salt, tautomer or stereoisomer of the compound as a drug carrier. wherein, The method comprises the following steps: Ra is selected from C 3-8 alkylene, 1-3 carbon atoms in said alkylene are optionally replaced by -O-, -S-, -NH-, with the proviso that said -O-, -S-, -NH- are not directly attached to a X group or a R1R2N- group, said -O-, -S-, -NH- groups are not directly attached to each other; optionally one or more hydrogens in said Ra are replaced by Rb, said Rb is selected from C 1-8 alkyl; R3, R4, R5 are each independently selected from the group consisting of C 1-8 branched or straight chain alkyl; or one of (R3, R4, R5) together with its adjacent other one forms a 5- or 6-membered heterocyclic ring; optionally said 5- or 6-membered heterocyclic ring is substituted by one or more R6 selected from the group consisting of C 1-6 branched or straight chain alkyl; R1 and R2 are each independently selected from -H and C. 2-8 Straight-chain or branched alkyl groups, wherein C 2-8 The methylene groups contained in the straight-chain or branched alkyl groups are optionally replaced by one or more -S-, -O-, or -NH- groups, and one or more hydrogens in R1 and R2 are optionally replaced by -COOR7, -OCOR7, COSR7, -S COR7, -CONR7R8, or -NR7COR8, wherein R7 and R8 are each independently selected from -H, C 1-20 Branched or straight-chain alkyl groups, C 1-20 Branched or straight-chain alkenyl groups, C 1-20 It has a branched or straight-chain alkynyl group, and R1 and R2 are not both -H.

2. The phosphatidylcholine compound of the structure of Formula (I) or Formula (II), a pharmaceutically acceptable salt, tautomer, or stereoisomer of the compound according to claim 1, wherein S1. reacting hydroxyethyl methacrylate and β-mercaptoethylamine as raw materials at room temperature in a solvent to obtain an intermediate M1; the reaction formula is as follows: ； ； ； ； ； ； ； ； ； 。 3. The phosphatidylcholine compound of the structure of Formula (I) or Formula (II), a pharmaceutically acceptable salt, tautomer, or stereoisomer of the compound of claim 1, wherein has a structure as described below: ； ； ； ； ； ； ； ； ； 。 4. The phosphatidylcholine compound of the structure of Formula (I) or Formula (II), the pharmaceutically acceptable salt, tautomer or stereoisomer of the compound according to claim 1, characterized in that S3. reacting the intermediate M2 and 2-chloro-2-oxo-1,3,2-dioxaphospholane to obtain an intermediate M3; the reaction formula is as follows: 。 The definitions of R1, R2, R3, R4 and R5 are as described in claim 4.

5. The phosphatidylcholine compound of formula (I) according to claim 1, a pharmaceutically acceptable salt of the compound, a tautomer or a stereoisomer, characterized in that... In step S1, the molar ratio of SM1 to SM2 is 1:1-1.

2. 、 、 、 、 、 、 、 、 、 、 、 、 、 、 。 In step S1, the reaction temperature is between 10-30°C, and the reaction time is 3-6 hours.

7. A process for the preparation of a phosphatidylcholine compound of the structure of Formula (la), a pharmaceutically acceptable salt, tautomer or stereoisomer of said compound according to claim 4, characterized in that In step S1, the solvent is ethanol, and the volume ratio of hydroxyethyl methacrylate to the solvent is 1:5-10 (g / ml). In step S1, the post-treatment operation is as follows: the reaction solution is concentrated under reduced pressure to remove the solvent to obtain a colorless oily crude product, which is directly used in the next step without further purification. ； S2. Intermediate M1 and carrying out an addition reaction or a substitution reaction to obtain intermediate M2; the reaction formula is as follows: ， said R9is selected from the group consisting of C 0-6 straight or branched chain alkyl, one or more hydrogens of said R9being optionally replaced by -COOR7, -OCOR7, -CONR7R8, -NR7COR8, said R7, R8being each independently selected from the group consisting of -H, C 1-20 branched or straight chain alkyl, C 1-20 branched or straight chain alkenyl, C 1-20 branched or straight chain alkynyl, In step S2, the molar ratio of M1 to SM3 is between 1:2.2-3. ； S4. intermediate M3 and to obtain the phosphatidylcholine compound shown in formula I; the reaction formula is as follows: ； In step S2, a solvent-free reaction is adopted, the reaction temperature is between 70-80°C, and the time is between 40-60 hours.

8. A process for the preparation of a phosphatidylcholine compound of the structure of Formula (la), a pharmaceutically acceptable salt, tautomer or stereoisomer of said compound according to claim 7, characterized in that In step S2, 0.05 equivalent of BHT is added as a reaction stabilizer.

9. A method of preparing a phosphatidylcholine compound of the structure of Formula (la), a pharmaceutically acceptable salt, tautomer, or stereoisomer of the compound according to claim 7, wherein In step S2, 0.05 equivalent of a catalyst is added, and the catalyst is acetic acid or boric acid.

10. A method of preparing a phosphatidylcholine compound of the structure of Formula (la), a pharmaceutically acceptable salt, tautomer, or stereoisomer of the compound of claim 7, wherein In step S3, the reaction occurs under the action of an acid-binding agent, the molar ratio of M2 to SM4 to the acid-binding agent is 1:2:2.5, and the acid-binding agent is selected from organic basic or inorganic alkali.

11. A method of preparing a phosphatidylcholine compound of the structure of Formula (la), a pharmaceutically acceptable salt, tautomer, or stereoisomer of said compound according to claim 7, wherein In step S3, the reaction is carried out in an organic inert aprotic solvent.

12. The method for preparing the phosphatidylcholine compound of formula (Ia) according to claim 7, a pharmaceutically acceptable salt of the compound, a tautomer or a stereoisomer, characterized in that... In step S3, the reaction temperature is between 0-10°C, and the time is 1-3 hours.

13. The method for preparing the phosphatidylcholine compound of formula (Ia) according to claim 7, a pharmaceutically acceptable salt of the compound, a tautomer or a stereoisomer, characterized in that... In step S4, the reaction is carried out in a solvent selected from one or more of dry acetonitrile, tetrahydrofuran, DMF, DMSO, acetone and dioxane, and the mass volume ratio of the substrate to the solvent is 1:5-10 (g / ml).

14. The method for preparing the phosphatidylcholine compound of formula (Ia) according to claim 7, a pharmaceutically acceptable salt of the compound, a tautomer or a stereoisomer, characterized in that... In step S4, the reaction temperature is between 50-55°C, and the time is 20-50 hours.

15. A method for preparing a phosphatidylcholine compound of formula (Ia) according to claim 7, a pharmaceutically acceptable salt of the compound, a tautomer or a stereoisomer, characterized in that... In step S4, the post-treatment operation is as follows: the reaction solution is concentrated under reduced pressure to remove the solvent to obtain a light yellow oily crude product, which is directly purified by column chromatography to obtain the target product.

16. The method for preparing a phosphatidylcholine compound of formula (Ia) according to claim 7, a pharmaceutically acceptable salt of the compound, a tautomer or a stereoisomer, characterized in that, ​ 17. A method for preparing a phosphatidylcholine compound of formula (Ia) according to claim 7, a pharmaceutically acceptable salt of the compound, a tautomer or a stereoisomer, characterized in that... ​ 18. A method for preparing a phosphatidylcholine compound of formula (Ia) according to claim 7, a pharmaceutically acceptable salt of the compound, a tautomer or a stereoisomer, characterized in that... ​ 19. A method for preparing a phosphatidylcholine compound of formula (Ia) according to claim 7, a pharmaceutically acceptable salt of the compound, a tautomer or a stereoisomer, characterized in that... In the step S4, M3 and are in a molar ratio of 1 : 1.5-5.

20. A method for preparing a phosphatidylcholine compound of formula (Ia) according to claim 7, a pharmaceutically acceptable salt of the compound, a tautomer or a stereoisomer, characterized in that... ​ 21. A method of preparing a phosphatidylcholine compound of the structure of Formula (la), a pharmaceutically acceptable salt, tautomer, or stereoisomer of said compound according to claim 7, wherein ​ 22. The method for preparing the phosphatidylcholine compound of formula (Ia) according to claim 7, a pharmaceutically acceptable salt of the compound, a tautomer or a stereoisomer, characterized in that... ​ 23. A composition characterized in that The composition comprises the phosphatidylcholine compound of the structure of formula (I) or formula (II), the pharmaceutically acceptable salt, tautomer or stereoisomer of the compound according to any one of claims 1-5.

24. The composition of claim 23, wherein The composition further comprises other lipids.

25. The composition of claim 23, wherein The other lipid is selected from one or more of cholesterols, DMG-PEG 2000.

26. The composition of any one of claims 24-25, wherein The other lipid is selected from .

27. The composition of any one of claims 24-25, wherein The other lipids are selected from cholesterol.

28. The composition of any one of claims 24-25, wherein The other lipids are selected from DMG-PEG2000.

29. The composition of any one of claims 24-25, wherein The other lipid is selected from the group consisting of a combination of cholesterol and DMG-PEG 2000.

30. The composition of claim 29, wherein The molar percentage of the other lipids, the phosphatidylcholine compound of the structure of formula (I) or formula (II), cholesterol and DMG-PEG2000 is (40%-60%):(5%-15%):(22%-54.5%):(0.5%-1.5%).

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