A class of photosensitizer-containing lipid-like molecules and uses thereof

Abstract

The application relates to a class of lipid molecules containing photosensitizers and an application thereof, and belongs to the fine chemical field. The series of molecules are connected with photosensitizers with a photodynamic treatment effect and lipid molecules, belong to a class of novel bifunctional material molecules. The molecules contain lipid side chains, have the performance of combining RNA / DNA and other base sequences and self-assembling into nanoparticles, become high-quality RNA / DNA delivery carrier materials; and the photosensitizer part in the molecules can generate singlet oxygen to induce tumor cell death under light source irradiation. The series of molecules have the RNA / DNA delivery performance and the ability of photodynamic damage to tumor cells, can efficiently connect gene therapy and photodynamic therapy.

Description

Technical Field

[0001] This invention relates to a class of small molecule lipid nanomaterials for delivering nucleic acid drugs such as DNA or RNA to living cells and living organisms, and particularly to a class of lipid molecules containing photosensitizers and their applications, which belong to the field of fine chemicals. Background Technology

[0002] Gene therapy is a method of treating diseases by delivering genes (mRNA, DNA, or siRNA, etc.) into the patient's body as drugs. Gene therapy is a treatment strategy that addresses gene defects at their source. The genes delivered into cells treat diseases by directly expressing functional protein molecules, interfering with normal gene expression, or correcting genomic defects. However, DNA or RNA-based gene drugs are phosphate polymers, possessing strong electronegativity and high hydrophilicity, making them easily degraded by biological enzymes and unable to directly enter cells to exert their effects like conventional drugs. Therefore, the safe and effective delivery of gene drugs into cells or tissues in vitro or in vivo is crucial for the success of gene therapy.

[0003] Lipid molecules typically consist of a polyamine core and hydrophobic fatty acid chains. Rational modification of the polyamine core and hydrophobic side chains can regulate the properties of lipid molecules. The easily ionized polyamine core binds to strongly electronegative DNA or RNA via electrostatic forces, self-assembling into nanoparticles in aqueous solution due to hydrophilic-hydrophobic interactions. Adding auxiliary materials such as cholesterol, phospholipids, and polyethylene glycol (PEG) further stabilizes these nanoparticles. Lipophilic nanoparticles effectively protect gene-editing drugs from enzymatic degradation and facilitate endocytosis into cells. Furthermore, the introduction of PEG provides a degree of stability to the nanoparticle matrix, reducing particle aggregation and the probability of clearance by mononuclear macrophages in the circulatory system. In addition, the amino core in the material can be protonated in the acidic environment of endosomes and lysosomes, promoting endosome escape and releasing gene-editing drugs, and can be degraded after release.

[0004] Lipid-like molecular materials have seen rapid development in recent years due to their well-defined structures and high designability, and significant applications have been reported in the field of cancer treatment. These materials offer advantages such as high cellular uptake, low cytotoxicity, and ease of structural modification, allowing for the cascading of gene therapy and photodynamic therapy through chemical structure design. Therefore, the development of multifunctional lipid molecules is of significant value in the fields of fine chemicals and biomedicine. Summary of the Invention

[0005] This application is a divisional application of Chinese patent application No. 2023108329976, filed on July 7, 2023.

[0006] To achieve the integration of gene therapy and photodynamic therapy, this invention provides a series of highly biocompatible functionalized lipid molecules.

[0007] The technical solution adopted in this invention is: to provide a compound of formula I,

[0008]

[0009] Among them, R0 is independent of each other. , , ;

[0010] R1 is independent of each other. , , , , , , or ;

[0011] n are independent integers from 1 to 5.

[0012] R2 is independent of each other. , , C6-C 20 saturated alkane group or C6-C 20 The unsaturated alkane groups, wherein each R3 is independently C4-C6. 16 saturated alkane group, C4-C 16 unsaturated alkane groups or R4 is independent of each other. , or Each of p is an independent integer from 1 to 18;

[0013] m is 1 or 2;

[0014] The photosensitizer is selected from phthalocyanine, cyanine dyes, fluorescein, porphyrin, or doxorubicin.

[0015] The application of the compound in the preparation of nanomaterials for delivering nucleic acid drugs.

[0016] The application of the compound in the preparation of photosensitizer materials.

[0017] The application of the compound in the preparation of a dual-mode synergistic therapeutic agent combining gene therapy and photodynamic therapy.

[0018] The beneficial effects of this invention are as follows: This series of molecules combines photosensitizers with photodynamic therapeutic effects with lipid molecules via ester or amide bonds, belonging to a novel class of bifunctional material molecules. This series of materials can deliver gene-based drugs such as RNA / DNA. The photosensitizers linked by ester or amide bonds in the molecules can generate biotoxic singlet oxygen and other reactive oxygen species under excitation by a light source of a specific wavelength, thereby damaging tumor cells. The number of carbon atoms between the two tertiary amines in the amino core can regulate the overall basicity of the molecule, and the number of carbon atoms between the drug molecule and the amino core can regulate the overall hydrophilicity / hydrophobicity of the molecule; both affect the binding ability of lipid molecules to the base sequence. The advantage of this design series is that the molecules possess both RNA / DNA delivery performance and the ability to damage tumor cells via photodynamic therapy, enabling efficient tandem gene therapy and photodynamic therapy. The molecule contains lipid-like side chains, which enable it to bind to base sequences such as RNA / DNA and self-assemble into nanoparticles, making it a high-quality RNA / DNA delivery carrier material. The photosensitizer portion can generate singlet oxygen under light irradiation to induce tumor cell death. The series of materials can serve as a universal delivery platform for integrating phototherapy and immunotherapy. Attached Figure Description

[0019] Figure 1 This is a graph showing the cytotoxicity data for G1.

[0020] Figure 2 This is a graph showing the cytotoxicity data for G15.

[0021] Figure 3 This is an imaging diagram of the G1-induced intracellular singlet oxygen production capacity experiment.

[0022] Figure 4 This is an imaging image of the G15-induced intracellular singlet oxygen production capacity experiment.

[0023] Figure 5 This is a cell experiment imaging image of G1.

[0024] Figure 6 This is a cell experiment imaging image of G15.

[0025] Figure 7 This is a confocal image of G1 delivering GFP mRNA to living cells. Detailed Implementation

[0026] The present invention is illustrated by the following embodiments, but is not limited thereto, wherein, unless otherwise stated, all fractions and percentages are by weight.

[0027] This invention relates to a class of biodegradable small-molecule lipid molecules containing photosensitizers. The photosensitizer molecules are bound to lipid molecules via ester or amide groups. The series of molecules retains the structural characteristics of the tertiary amine core and lipophilic fatty chains of lipid molecules, enabling them to form nanoparticles with base sequences and efficiently deliver the base sequences to cells and living organisms. While achieving transmembrane delivery of gene drugs, the photosensitizers linked by ester or amide bonds in the molecules can generate biotoxic singlet oxygen and other reactive oxygen species, thereby damaging tumor cells and achieving synergistic therapy. Specifically, in the structure of the embodiment, I... - It is an iodide ion.

[0028] Compounds of Formula I or their salts:

[0029]

[0030] Among them, R0 is independent of each other. , , ;

[0031] R1 is independent of each other. , , , , , , or ;

[0032] n are independent integers from 1 to 5.

[0033] R2 is independent of each other. , , C6-C 20 saturated alkane group or C6-C 20 The unsaturated alkane groups, wherein each R3 is independently C4-C6. 16 saturated alkane group, C4-C 16 unsaturated alkane groups or R4 is independent of each other. , or Each of p is an independent integer from 1 to 18;

[0034] m is 1 or 2;

[0035] The photosensitizer is selected from phthalocyanine, cyanine dyes, fluorescein, porphyrin, or doxorubicin.

[0036] Specifically, X is independent of each other. , , , , , , , , or ;

[0037] Among them, Y - Each anion is independent and selected from BF4. - Cl - ,Br - I - NO 3- SO4 2- ClO4 - CH3COO - CH3SO3 - CF3SO3 - The number of negative charges carried by the anions is equal to the number of positive charges carried by the cations.

[0038] A class of photosensitizer-containing degradable small-molecule lipid molecules, having formula I1-I 10 Structure:

[0039]

[0040]

[0041]

[0042]

[0043]

[0044] Among them, R0, R1, R 2、 Y - The definition is the same as that in formula I.

[0045] In some specific compounds, R1 is independently... , , , , , , , , or .

[0046] In some specific compounds, R2 is independently... , , C8-C 18 Saturated alkane group or C8-C 18The unsaturated alkane groups, wherein each R3 is independently C6-C6. 14 Saturated alkane group, C6-C 14 unsaturated alkane groups or R4 is independent of each other. , or Each p is an independent integer between 0 and 12.

[0047] In some specific compounds, R0 is independently... , , or .

[0048] In some specific compounds, R2 is independently... , , , , , , , , , , , , , , , , , , , , , , , , , , , or ;

[0049] Among them, R3 is independent of each other. , , , , , , , , , , , , , , , , , , , , , , or ;

[0050] R4 is independent of each , or .

[0051] In some specific compounds, R2 is independently... , , , , , , , , , , , , , , , , , , , , or ;

[0052] Among them, R3 is independent of each other. , , , , , , , , , , , , , , , or .

[0053] R4 is independent of each , or .

[0054] Specific preparation methods for this type of compound:

[0055] Lipid-like molecules containing ester groups: Their synthesis begins with alkylamines containing two or more primary amines, followed by unilateral amino protection and nucleophilic substitution of bromoalkane to obtain an intermediate containing a single hydroxyl group; after deesterification protection with trifluoroacetic acid, lipid-like intermediates containing three long ester chains or long carbon chains and one free hydroxyl group are obtained through Mac addition or reductive amination; finally, the target compound is obtained through a one-step esterification condensation reaction.

[0056] Lipid molecules containing amide groups: Their synthesis begins with alkylamines containing two or more primary amines, followed by unilateral amino protection and nucleophilic substitution of bromoalkane to obtain intermediates containing a single hydroxyl group; after deesterification protection with trifluoroacetic acid, lipid intermediates containing three long ester chains or long carbon chains are obtained through Mac addition or reductive ammoniation; then, lipid intermediates containing a single amino group in the side chain are obtained through azidation and reduction reactions; finally, the target compound is obtained through a one-step amidation condensation reaction.

[0057] The specific embodiments of the present invention are described in detail below with reference to the technical solutions:

[0058] Example 1

[0059]

[0060] 2 g of the starting compound N-(2-hydroxyethyl)-1,3-propanediamine (16.9 mmol) was dissolved in 50 mL of tetrahydrofuran. 15 g of compound A1 and NaBH(AcO)3 (84.6 mmol) were added with stirring. The reaction was carried out at room temperature for 24 h. The solvent was removed by vacuum distillation. The crude product was purified by silica gel column chromatography to obtain compound B1.

[0061] 1.5 g of compound B1 was dissolved in 10 mL of dry CH2Cl2, followed by the addition of 0.7 mL of triethylamine and a catalytic amount of DMAP. The mixture was then added dropwise with stirring at 0 °C in 10 mL of dichloromethane containing 700 mg of p-toluenesulfonyl chloride. After the reaction was complete, the solvent was removed by vacuum distillation, and the mixture was redissolved in 10 mL of DMF, followed by the addition of 300 mg of NaN3. The reaction was continued at 80 °C for 5 h. After the reaction was complete, the organic phase, after the large amount of dichloromethane dissolved, was washed repeatedly with 20 mL of saturated brine, dried over anhydrous sodium sulfate, and the solvent was removed by rotary evaporation under reduced pressure. The crude product was purified by silica gel column chromatography to obtain compound C1.

[0062] Under argon protection, 200 mg of compound C1 was dissolved in 5 mL of ultra-dry tetrahydrofuran, and 35 mg of lithium aluminum hydride was added at 0 °C. The reaction was stirred at room temperature for 30 min, and quenched by the slow addition of 2 mL of water and 2 mL of 15% sodium hydroxide aqueous solution. The organic phase was washed repeatedly with 20 mL of saturated brine, dried over anhydrous sodium sulfate, and the solvent was removed under reduced pressure. The crude product was purified by silica gel column chromatography to obtain compound D1.

[0063] 20 mg of compound E1 was dissolved in a mixed solution of 3 mL CH2Cl2 and 1 mL MeCN. 107 mg of 2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate and 39 μL of triethylamine were added sequentially to the mixed solution. The mixture was reacted at 0 °C in an ice bath for 30 min, and then 52 mg of compound D1 was added. The mixture was stirred overnight at room temperature. The solvent was removed by rotary evaporation under reduced pressure. The product was purified by silica gel column chromatography to obtain 45.45 mg of the target compound G1, with a yield of 93%. The product structure was identified by HRMS, HRMS [M+H]+: 1916.7589.

[0064] Example 2

[0065]

[0066] 2 g of the starting compound hydroxyethyl ethylenediamine (19.2 mmol) was dissolved in 50 mL of tetrahydrofuran. 27 g of compound A2 and NaBH(AcO)3 (96 mmol) were added with stirring. The reaction was carried out at room temperature for 24 h. The solvent was removed by vacuum distillation. The crude product was purified by silica gel column chromatography to obtain compound B2.

[0067] 1.5 g of compound B2 was dissolved in 10 mL of dry CH2Cl2, followed by the addition of 0.5 mL of triethylamine and a catalytic amount of DMAP. The mixture was then added dropwise with stirring at 0 °C in 10 mL of dichloromethane containing 471 mg of p-toluenesulfonyl chloride. After the reaction was complete, the solvent was removed by vacuum distillation, and the mixture was redissolved in 10 mL of DMF. Then, 215 mg of NaN3 was added, and the reaction was carried out at 80 °C for 5 h. After the reaction was complete, the organic phase was washed repeatedly with small amounts of 20 mL of saturated brine, dried over anhydrous sodium sulfate, and the solvent was removed by rotary evaporation under reduced pressure. The crude product was purified by silica gel column chromatography to obtain compound C2.

[0068] Under argon protection, 1 g of compound C2 was dissolved in 20 mL of ultra-dry tetrahydrofuran, and 122 mg of lithium aluminum hydride was added at 0 °C. The reaction was stirred at room temperature for 30 min, and quenched by the slow addition of 5 mL of water and 5 mL of 15% sodium hydroxide aqueous solution. The organic phase was washed repeatedly with 20 mL of saturated brine, dried over anhydrous sodium sulfate, and the solvent was removed under reduced pressure. The crude product was purified by silica gel column chromatography to obtain compound D2.

[0069] 100 mg of compound E2 was dissolved in a mixed solution of 6 mL CH2Cl2 and 2 mL MeCN. Then, 572 mg of 2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate and 0.2 mL of triethylamine were added sequentially to the mixed solution. The mixture was reacted at 0 °C in an ice bath for 30 min, followed by the addition of 685 mg of compound D2. The reaction was stirred overnight at room temperature. The solvent was removed by rotary evaporation under reduced pressure. The product was purified by silica gel column chromatography to obtain 232.19 mg of the target compound G2, with a yield of 70%. The product structure was identified by HRMS [M+H]. + :2564.0378.

[0070] Example 3

[0071]

[0072] 5.37 g of the starting compound N-methyl-2,2-diaminodiethylamine was dissolved in 55 mL of dichloromethane. Under stirring, 30 mL of a dichloromethane solution containing 5 g of Boc₂O was added dropwise over 3 hours. After the reaction was complete, the organic phase was washed three times successively with 20 mL of 1 mol / L sodium bicarbonate aqueous solution and 20 mL of saturated brine, dried over anhydrous magnesium sulfate, and the solvent was removed by vacuum evaporation to obtain the target compound A3.

[0073] 3.75 g of compound A3 was dissolved in 15 mL of DMF solution. 2.4 mL of triethylamine and 5 mL of DMF solution containing 1.2 g of 3-bromo-1-propanol were added with stirring. The mixture was reacted at room temperature for 24 h. The solvent was removed by vacuum distillation. The crude product was purified by silica gel column chromatography to obtain compound B3.

[0074] 3.48 mL of trifluoroacetic acid was added to a 500 mg solution of compound B3 in dichloromethane, and the mixture was stirred at room temperature for 1 h. The solvent was removed by vacuum evaporation, and the crude product containing C3 was directly used for the next reaction without purification.

[0075] 220 mg of the crude product of compound C3 was dissolved in 10 mL of tetrahydrofuran. The pH of the solution was adjusted to neutral by adding an appropriate amount of triethylamine. The reaction was carried out at room temperature for 30 min. Then, 1.86 g of compound D3 and NaBH(AcO)3 (6.28 mmol) were added, and the reaction was carried out at room temperature for 24 h. The solvent was removed by vacuum distillation, and the crude product was purified by silica gel column chromatography to obtain compound E3.

[0076] 235 mg of compound E3 was dissolved in a mixture of 2 mL dichloromethane and 2 mL DMF. Then, 182 mg of F3, 72 mg of EDC, and 3 mg of DMAP were added sequentially to the mixture. The mixture was stirred overnight at room temperature. The solvent was removed by rotary evaporation under reduced pressure. The product was purified by silica gel column chromatography to obtain 277 mg of the target compound G3, with a yield of 46%. The product structure was identified by HRMS [M+H]. + :2640.9111.

[0077] Example 4

[0078]

[0079] 11.98 g of the starting compound N,N-bis(2-aminoethyl)-N1,N2-dimethyl-1,2-ethylenediamine was dissolved in 55 mL of dichloromethane. Under stirring, 30 mL of a dichloromethane solution containing 7.5 g of Boc₂O was added dropwise over 3 hours. After the reaction was complete, the organic phase was washed three times successively with 20 mL of 1 mol / L sodium bicarbonate aqueous solution and 20 mL of saturated brine, dried over anhydrous sodium sulfate, and the solvent was removed by vacuum evaporation to obtain the target compound A4.

[0080] 7.90 g of compound A4 was dissolved in 25 mL of DMF solution. 4 mL of triethylamine and 5 mL of DMF solution containing 2 g of 3-bromo-1-propanol were added with stirring. The mixture was reacted at room temperature for 24 h. The solvent was removed by vacuum distillation. The crude product was purified by silica gel column chromatography to obtain compound B4.

[0081] Add 5.75 mL of trifluoroacetic acid to a 1 g solution of compound B4 in dichloromethane, stir at room temperature for 1 h, remove the solvent under reduced pressure, and proceed directly to the next step of the reaction without purification.

[0082] 475 mg of the crude product of compound C4 was dissolved in 10 mL of tetrahydrofuran. The pH of the solution was adjusted to neutral by adding an appropriate amount of triethylamine. The reaction was carried out at room temperature for 30 min. Then, 2.23 g of compound D4 and NaBH(AcO)3 (10.22 mmol) were added, and the reaction was carried out at room temperature for 24 h. The solvent was removed by vacuum distillation, and the crude product was purified by silica gel column chromatography to obtain compound E4.

[0083] 687 mg of compound E4 was dissolved in a mixture of 3 mL dichloromethane and 3 mL DMF. 753 mg F4, 241 mg EDC and 9.5 mg DMAP were added to the mixture sequentially. The mixture was stirred overnight at room temperature. The solvent was removed by rotary evaporation under reduced pressure. The product was purified by silica gel column chromatography to obtain 1.034 g of the target compound G4, with a yield of 43%. The structure of the product was identified by HRMS, HRMS [M+H]+: 2464.7458.

[0084] Example 5

[0085]

[0086] 5.51 g of the starting compound 1,4-bis(3-aminopropyl)piperazine was dissolved in 30 mL of dichloromethane. Under stirring, 15 mL of a dichloromethane solution containing 3 g of Boc₂O was added dropwise over 3 hours. After the reaction was complete, the organic phase was washed three times successively with 15 mL of 1 mol / L sodium bicarbonate aqueous solution and 15 mL of saturated brine, dried over anhydrous sodium sulfate, and the solvent was removed by vacuum evaporation to obtain the target compound A5.

[0087] 2.55 g of compound A5 was dissolved in 20 mL of DMF solution, and 1.18 mL of triethylamine and 5 mL of DMF solution containing 650 mg of bromoacetic acid were added with stirring. The mixture was reacted at room temperature for 24 h, and the solvent was removed by vacuum distillation. The crude product was purified by silica gel column chromatography to obtain compound B5.

[0088] 3 mL of trifluoroacetic acid was added to a dichloromethane solution containing 584 mg of compound B5, and the mixture was stirred at room temperature for 1 h. The solvent was removed by vacuum evaporation, and the crude product containing C5 was directly used for the next reaction without purification.

[0089] 400 mg of the crude product of compound C5 was dissolved in 10 mL of tetrahydrofuran. The pH of the solution was adjusted to neutral by adding an appropriate amount of triethylamine. The reaction was carried out at room temperature for 30 min. Then, 2.51 g of compound D5 and NaBH(AcO)3 (7.34 mmol) were added, and the reaction was carried out at room temperature for 24 h. The solvent was removed by vacuum distillation, and the crude product was purified by silica gel column chromatography to obtain compound E5.

[0090] 185.4 mg of compound E5 was dissolved in a mixture of 1 mL dichloromethane and 1 mL DMF. 248 mg of F5, 115 mg of EDC and 2 mg of DMAP were added to the mixture sequentially. The mixture was stirred overnight at room temperature. The solvent was removed by rotary evaporation under reduced pressure. The product was purified by silica gel column chromatography to obtain 172 mg of the target compound G5, with a yield of 35%. The structure of the product was identified by HRMS, HRMS [M+H]+: 3302.5540.

[0091] Example 6

[0092]

[0093] 6.52 g of the starting compound 1,4-cyclohexylamine dimethylamine was dissolved in 50 mL of dichloromethane. Under stirring, 30 mL of a dichloromethane solution containing 5 g of Boc₂O was added dropwise over 3 hours. After the reaction was complete, the organic phase was washed three times successively with 20 mL of 1 mol / L sodium bicarbonate aqueous solution and 20 mL of saturated brine, dried over anhydrous sodium sulfate, and the solvent was removed by vacuum evaporation to obtain the target compound A6.

[0094] 4.53 g of compound A6 was dissolved in 20 mL of DMF solution, and 2.60 mL of triethylamine and 10 mL of DMF solution containing 1.3 g of 3-bromo-1-propanol were added with stirring. The reaction was carried out at room temperature for 24 h, and the solvent was removed by vacuum distillation. The crude product was purified by silica gel column chromatography to obtain compound B6.

[0095] 11.53 mL of trifluoroacetic acid was added to a dichloromethane solution of 1.81 g of compound B6, and the mixture was stirred at room temperature for 1 h. The solvent was removed by vacuum evaporation, and the crude product containing C6 was directly used for the next reaction without purification.

[0096] 1.21 g of the crude product of compound C6 was dissolved in 30 mL of tetrahydrofuran. The pH of the solution was adjusted to neutral by adding an appropriate amount of triethylamine. The reaction was carried out at room temperature for 30 min. Then, 12.04 g of compound D6 and NaBH(AcO)3 (30.2 mmol) were added. The reaction was carried out at room temperature for 24 h. The solvent was removed by vacuum distillation. The crude product was purified by silica gel column chromatography to obtain compound E6.

[0097] 995 mg of compound E6 was dissolved in a mixture of 3 mL dichloromethane and 3 mL DMF. 440 mg of F6, 230 mg of EDC and 9 mg of DMAP were added to the mixture sequentially. The mixture was stirred overnight at room temperature. The solvent was removed by rotary evaporation under reduced pressure. The product was purified by silica gel column chromatography to obtain 593 mg of the target compound G6, with a yield of 42%. The structure of the product was identified by HRMS, HRMS [M+H]+: 1926.3701.

[0098] Example 7

[0099]

[0100] 2.95 g of the starting compound 1,8-diamino-3,6-dioxaoctane was dissolved in 30 mL of dichloromethane. Under stirring, 15 mL of a dichloromethane solution containing 2.18 g of Boc₂O was added dropwise over 3 hours. After the reaction was complete, the organic phase was washed three times successively with 20 mL of 1 mol / L sodium bicarbonate aqueous solution and 20 mL of saturated brine, dried over anhydrous magnesium sulfate, and the solvent was removed by vacuum evaporation to obtain the target compound A7.

[0101] 2.40 g of compound A6 was dissolved in 15 mL of DMF solution, and 1.27 mL of triethylamine and 2 mL of DMF solution containing 700 mg of bromoacetic acid were added with stirring. The reaction was carried out at room temperature for 24 h, and the solvent was removed by vacuum distillation. The crude product was purified by silica gel column chromatography to obtain compound B7.

[0102] 3 mL of trifluoroacetic acid was added to a dichloromethane solution containing 523.2 mg of compound B7, and the mixture was stirred at room temperature for 1 h. The solvent was removed by vacuum evaporation, and the crude product containing C7 was directly used for the next reaction without purification.

[0103] 344.6 mg of the crude product of compound C7 was dissolved in 10 mL of tetrahydrofuran solution, 317 mg of triethylamine was added, and the reaction was carried out at room temperature for 30 min. Then, 1.66 g of compound D7 and NaBH(OAc)3 (7.82 mmol) were added, and the reaction was carried out at room temperature for 24 h. The solvent was removed by vacuum distillation, and the crude product was purified by silica gel column chromatography to obtain compound E7.

[0104] 186.8 mg of compound E7 was dissolved in a mixture of 3 mL dichloromethane and 3 mL DMF. 148.32 mg of F7, 67.65 mg of EDC and 3 mg of DMAP were added to the mixture sequentially. The mixture was stirred overnight at room temperature. The solvent was removed by rotary evaporation under reduced pressure. The product was purified by silica gel column chromatography to obtain 130 mg of the target compound G7, with a yield of 39%. The structure of the product was identified by HRMS, HRMS [M+H]+: 1520.0195.

[0105] Example 8

[0106]

[0107] 1.5 g of the starting compound N-(2-hydroxyethyl)-1,3-propanediamine was dissolved in 50 mL of tetrahydrofuran. 16.8 g of compound A8 and NaBH(AcO)3 (63.46 mmol) were added with stirring. The reaction was carried out at room temperature for 24 h. The solvent was removed by vacuum distillation. The crude product was purified by silica gel column chromatography to obtain compound B8.

[0108] 2.35 g of compound B8 was dissolved in 20 mL of dry CH2Cl2, followed by the addition of 550 mg of triethylamine and a catalytic amount of DMAP. Under stirring at 0 °C, 10 mL of dichloromethane solution containing 778 mg of p-toluenesulfonyl chloride was added dropwise. After the reaction was complete, the solvent was removed by vacuum distillation, and the mixture was redissolved in 20 mL of DMF, followed by the addition of 353.83 mg of NaN3. The reaction was carried out at 80 °C for 5 h. After the reaction was complete, the organic phase, after the large amount of dichloromethane dissolved, was washed repeatedly with 20 mL of saturated brine, dried over anhydrous sodium sulfate, and the solvent was removed by rotary evaporation under reduced pressure. The crude product was purified by silica gel column chromatography to obtain compound C8.

[0109] Under argon protection, 283.19 mg of compound C8 was dissolved in 5 mL of ultra-dry tetrahydrofuran, and 36.29 mg of lithium aluminum hydride was added at 0 °C. The reaction was stirred at room temperature for 30 min, and quenched by the slow addition of 2 mL of water and 2 mL of 15% sodium hydroxide aqueous solution. The organic phase was washed repeatedly with 20 mL of saturated brine, dried over anhydrous sodium sulfate, and the solvent was removed under reduced pressure. The crude product was purified by silica gel column chromatography to obtain compound D8.

[0110] 81.3 mg of compound E8 was dissolved in a mixed solution of 3 mL CH2Cl2 and 1 mL MeCN. Then, 516.34 mg of 2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate and 137.41 mg of triethylamine were added sequentially to the mixed solution. The mixture was reacted at 0 °C in an ice bath for 30 min, followed by the addition of 585.65 mg of compound D8. The reaction was stirred overnight at room temperature. The solvent was removed by rotary evaporation under reduced pressure. The product was purified by silica gel column chromatography to obtain 150 mg of the target compound G8, with a yield of 48%. The product structure was identified by HRMS [M+H]. + :2287.0170.

[0111] Example 9

[0112]

[0113] 9.67 g of the starting compound N-methyl-2,2-diaminodiethylamine was dissolved in 55 mL of dichloromethane. Under stirring, 30 mL of a dichloromethane solution containing 9 g of Boc₂O was added dropwise over 3 hours. After the reaction was complete, the organic phase was washed three times successively with 20 mL of 1 mol / L sodium bicarbonate aqueous solution and 20 mL of saturated brine, dried over anhydrous magnesium sulfate, and the solvent was removed by vacuum evaporation to obtain the target compound A9.

[0114] 5 g of compound A9 was dissolved in 15 mL of DMF solution, and 2.33 g of triethylamine and 5 mL of DMF solution containing 1.60 g of 3-bromo-1-propanol were added with stirring. The reaction was carried out at room temperature for 24 h, and the solvent was removed by vacuum distillation. The crude product was purified by silica gel column chromatography to obtain compound B9.

[0115] 2.66 mL of trifluoroacetic acid was added to a dichloromethane solution containing 385.49 mg of compound B9, and the mixture was stirred at room temperature for 1 h. The solvent was removed by vacuum evaporation, and the crude product containing C9 was directly used for the next reaction without purification.

[0116] 70.5 mg of the crude product of compound C9 was dissolved in 5 mL of tetrahydrofuran solution, 81.4 mg of triethylamine was added, and the reaction was carried out at room temperature for 30 min. Then, 515.62 mg of compound D9 and NaBH(OAc)3 (2.01 mmol) were added, and the reaction was carried out at room temperature for 24 h. The solvent was removed by vacuum distillation, and the crude product was purified by silica gel column chromatography to obtain compound E9.

[0117] 190 mg of compound E9 was dissolved in a mixture of 2 mL dichloromethane and 2 mL DMF. 605.41 mg F9, 104.84 mg EDC and 4 mg DMAP were added to the mixture sequentially. The mixture was stirred overnight at room temperature. The solvent was removed by rotary evaporation under reduced pressure. The product was purified by silica gel column chromatography to obtain 327.9 mg of the target compound G9, with a yield of 42%. The structure of the product was identified by HRMS, HRMS [M+H]+: 2318.8307.

[0118] Example 10

[0119]

[0120] 11.11 g of the starting compound was dissolved in 65 mL of dichloromethane. Under stirring, 30 mL of a dichloromethane solution containing 15.93 g of Boc₂O was added dropwise over 3 hours. After the reaction was complete, the organic phase was washed three times successively with 20 mL of 1 mol / L sodium bicarbonate aqueous solution and 20 mL of saturated brine, dried over anhydrous magnesium sulfate, and the solvent was removed by vacuum evaporation to obtain the target compound A10.

[0121] 5.25 g of compound A10 was dissolved in 15 mL of DMF solution, and 3.01 g of triethylamine and 5 mL of DMF solution containing 2.07 g of 3-bromo-1-propanol were added with stirring. The reaction was carried out at room temperature for 24 h, and the solvent was removed by vacuum distillation. The crude product was purified by silica gel column chromatography to obtain compound B10.

[0122] 2.65 mL of trifluoroacetic acid was added to a dichloromethane solution containing 325.05 mg of compound B10, and the mixture was stirred at room temperature for 1 h. The solvent was removed by vacuum evaporation, and the crude product containing C10 was directly used for the next reaction without purification.

[0123] 52.85 mg of the crude product of compound C10 was dissolved in 5 mL of tetrahydrofuran solution, 200 mg of triethylamine was added, and the reaction was carried out at room temperature for 30 min. Then, 390.54 mg of compound D10 and NaBH(OAc)3 (1.97 mmol) were added, and the reaction was carried out at room temperature for 24 h. The solvent was removed by vacuum distillation, and the crude product was purified by silica gel column chromatography to obtain compound E10.

[0124] 281.29 mg of compound E10 was dissolved in a mixture of 2 mL dichloromethane and 2 mL DMF. 205.50 mg F10, 119.79 mg EDC and 4.71 mg DMAP were added to the mixture sequentially. The mixture was stirred overnight at room temperature. The solvent was removed by rotary evaporation under reduced pressure. The product was purified by silica gel column chromatography to obtain 228.77 mg of the target compound G10, with a yield of 47%. The structure of the product was identified by HRMS, HRMS [M+H]+: 1242.8283.

[0125] Example 11

[0126]

[0127] 2.85 g of the starting compound ethylenediamine was dissolved in 30 mL of dichloromethane. Under stirring, 15 mL of a dichloromethane solution containing 5.18 g of Boc₂O was added dropwise over 3 hours. After the reaction was complete, the organic phase was washed three times successively with 20 mL of 1 mol / L sodium bicarbonate aqueous solution and 20 mL of saturated brine, dried over anhydrous magnesium sulfate, and the solvent was removed by vacuum evaporation to obtain the target compound A11.

[0128] 2.24 g of compound A11 was dissolved in 10 mL of DMF solution, and 1.42 g of triethylamine and 5 mL of DMF solution containing 1.07 g of bromoacetic acid were added with stirring. The mixture was reacted at room temperature for 24 h, and the solvent was removed by vacuum distillation. The crude product was purified by silica gel column chromatography to obtain compound B11.

[0129] 2.69 mL of trifluoroacetic acid was added to a dichloromethane solution containing 328.44 mg of compound B11, and the mixture was stirred at room temperature for 1 h. The solvent was removed by vacuum evaporation, and the crude product containing C11 was directly used for the next reaction without purification.

[0130] 100 mg of the crude product of compound C11 was dissolved in 5 mL of tetrahydrofuran solution, 382.8 mg of triethylamine was added, and the reaction was carried out at room temperature for 30 min. Then, 863.8 mg of compound D11 and NaBH(OAc)3 (3.78 mmol) were added, and the reaction was carried out at room temperature for 24 h. The solvent was removed by vacuum distillation, and the crude product was purified by silica gel column chromatography to obtain compound E11.

[0131] 216.86 mg of compound E11 was dissolved in a mixture of 6 mL dichloromethane and 2 mL acetonitrile. 1.01 g of 2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate and 268.55 mg of triethylamine were added sequentially to the mixture. The reaction was carried out at 0 °C in an ice bath for 30 min, followed by the addition of 172.77 mg of compound F11. The reaction was stirred overnight at room temperature. The solvent was removed by rotary evaporation under reduced pressure. The product was purified by silica gel column chromatography to obtain 152.35 mg of the target compound G11, with a yield of 39%. The product structure was identified by HRMS [M+H]. + :1447.7286.

[0132] Example 12

[0133]

[0134] 30 g of the starting compound 1,3-propanediamine was dissolved in 150 mL of dichloromethane. Under stirring, 150 mL of a dichloromethane solution containing 15 g of Boc₂O was added dropwise over 3 hours. After the reaction was complete, the organic phase was washed three times successively with 30 mL of 1 mol / L sodium bicarbonate aqueous solution and 30 mL of saturated brine, dried over anhydrous magnesium sulfate, and the solvent was removed by vacuum evaporation to obtain the target compound A12.

[0135] 3.12 g of compound A12 was dissolved in 10 mL of DMF solution, and 1.81 g of triethylamine and 10 mL of DMF solution containing 2 g of 6-bromohexanoic acid were added with stirring. The reaction was carried out at room temperature for 24 h, and the solvent was removed by vacuum distillation. The crude product was purified by silica gel column chromatography to obtain compound B12.

[0136] 3.63 mL of trifluoroacetic acid was added to a 600 mg solution of compound B12 in dichloromethane, and the mixture was stirred at room temperature for 1 h. The solvent was removed by vacuum evaporation, and the crude product containing C12 was directly used for the next reaction without purification.

[0137] 410.17 mg of the crude product of compound C12 was dissolved in 10 mL of tetrahydrofuran. The pH of the solution was adjusted to neutral by adding an appropriate amount of triethylamine. The reaction was carried out at room temperature for 30 min. Then, 1.61 g of undecaldehyde and 9.46 mmol of NaBH(AcO)3 were added, and the reaction was carried out at room temperature for 24 h. The solvent was removed by vacuum distillation, and the crude product was purified by silica gel column chromatography to obtain compound D12.

[0138] Compound D12 was dissolved in 15 mL of tetrahydrofuran and 15 mL of methanol, and then 15 mL of 1.5 mol / L NaOH aqueous solution was added. The mixture was stirred at 80 °C for 4 h. After the reaction was completed, methanol, tetrahydrofuran, and some water were removed by rotary evaporation. The product was dissolved in 150 mL of dichloromethane, washed repeatedly with 20 mL of saturated brine, dried over anhydrous magnesium sulfate, and the solvent was removed by rotary evaporation under reduced pressure. The crude product containing E12 was directly used in the next reaction without purification.

[0139] 167.35 mg of compound E12 was dissolved in a mixture of 3 mL dichloromethane and 1 mL acetonitrile. 977.21 mg of 2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate and 260.07 mg of triethylamine were added sequentially to the mixture. The reaction was carried out at 0 °C in an ice bath for 30 min, followed by the addition of 139.69 mg of DOX F12. The reaction was stirred overnight at room temperature. The solvent was removed by rotary evaporation under reduced pressure. The product was purified by silica gel column chromatography to obtain 197.3 mg of the target compound G12, with a yield of 65%. The product structure was identified by HRMS [M+H]. + :1175.8324.

[0140] Example 13

[0141]

[0142] 6.44 g of the starting compound N-methyl-2,2-diaminodiethylamine was dissolved in 50 mL of dichloromethane. Under stirring, 30 mL of a dichloromethane solution containing 6 g of Boc₂O was added dropwise over 3 hours. After the reaction was complete, the organic phase was washed three times successively with 20 mL of 1 mol / L sodium bicarbonate aqueous solution and 20 mL of saturated brine, dried over anhydrous magnesium sulfate, and the solvent was removed by vacuum evaporation to obtain the target compound A13.

[0143] 4.22 g of compound A13 was dissolved in 15 mL of DMF solution, and 1.97 g of triethylamine and 5 mL of DMF solution containing 1.35 g of 3-bromo-1-propanol were added with stirring. The reaction was carried out at room temperature for 24 h, and the solvent was removed by vacuum distillation. The crude product was purified by silica gel column chromatography to obtain compound B13.

[0144] 8.82 mL of trifluoroacetic acid was added to a 1.27 g solution of compound B13 in dichloromethane, and the mixture was stirred at room temperature for 1 h. The solvent was removed by vacuum evaporation, and the crude product containing C13 was directly used for the next reaction without purification.

[0145] 175.6 mg of the crude product of compound C13 was dissolved in 10 mL of tetrahydrofuran solution, 202.76 mg of triethylamine was added, and the reaction was carried out at room temperature for 30 min. Then, 792.45 mg of compound D13 and 5.01 mmol of NaBH(OAc)3 were added, and the reaction was carried out at room temperature for 24 h. The solvent was removed by vacuum distillation, and the crude product was purified by silica gel column chromatography to obtain compound E13.

[0146] 350.97 mg of compound E13 was dissolved in a mixture of 2 mL dichloromethane and 2 mL DMF. Then, 207 mg of F13, 90.53 mg of EDC, and 3.56 mg of DMAP were added sequentially to the mixture. The mixture was stirred overnight at room temperature. The solvent was removed by rotary evaporation under reduced pressure. The product was purified by silica gel column chromatography to obtain 258.3 mg of the target compound G13, with a yield of 50%. The product structure was identified by HRMS: [M+H]+: 1961.4093.

[0147] Example 14

[0148]

[0149] 1.21 g of the starting compound ethylenediamine was dissolved in 30 mL of dichloromethane. Under stirring, 10 mL of a dichloromethane solution containing 2.18 g of Boc₂O was added dropwise over 3 hours. After the reaction was complete, the organic phase was washed three times successively with 20 mL of 1 mol / L sodium bicarbonate aqueous solution and 20 mL of saturated brine, dried over anhydrous magnesium sulfate, and the solvent was removed by vacuum evaporation to obtain the target compound A14.

[0150] 4.05 g of compound A11 was dissolved in 20 mL of DMF solution, and 2.56 g of triethylamine and 10 mL of DMF solution containing 2.29 g of 6-bromo-1-hexanol were added with stirring. The reaction was carried out at room temperature for 24 h, and the solvent was removed by vacuum distillation. The crude product was purified by silica gel column chromatography to obtain compound B14.

[0151] 14.25 mL of trifluoroacetic acid was added to a dichloromethane solution of 1.94 g of compound B14, and the mixture was stirred at room temperature for 1 h. The solvent was removed by vacuum evaporation, and the crude product containing C14 was directly used for the next reaction without purification.

[0152] 974 mg of the crude product of compound C14 was dissolved in 10 mL of tetrahydrofuran solution, 1.23 g of triethylamine was added, and the reaction was carried out at room temperature for 30 min. Then, 6.09 g of compound D14 and NaBH(OAc)3 (30.39 mmol) were added, and the reaction was carried out at room temperature for 24 h. The solvent was removed by vacuum distillation, and the crude product was purified by silica gel column chromatography to obtain compound E14.

[0153] 1.79 g of compound E14 was dissolved in a mixture of 5 mL dichloromethane and 5 mL DMF. 857.3 mg F14, 388.96 mg EDC, and 15.3 mg DMAP were added to the mixture sequentially. The mixture was stirred overnight at room temperature. The solvent was removed by rotary evaporation under reduced pressure. The product was purified by silica gel column chromatography to obtain 950.9 mg of the target compound G14, with a yield of 38%. The structure of the product was identified by HRMS, HRMS [M+H]+: 2241.5363.

[0154] Example 15

[0155]

[0156] 3.86 g of the starting compound 1,3-propanediamine was dissolved in 20 mL of dichloromethane. Under stirring, 20 mL of a dichloromethane solution containing 5.68 g of Boc₂O was added dropwise over 3 hours. After the reaction was complete, the organic phase was washed three times successively with 20 mL of 1 mol / L sodium bicarbonate aqueous solution and 20 mL of saturated brine, dried over anhydrous magnesium sulfate, and the solvent was removed by vacuum evaporation to obtain the target compound A15.

[0157] 3.06 g of compound A15 was dissolved in 10 mL of DMF solution, and 1.78 g of triethylamine and 10 mL of DMF solution containing 1.22 g of 3-bromo-1-propanol were added with stirring. The reaction was carried out at room temperature for 24 h, and the solvent was removed by vacuum distillation. The crude product was purified by silica gel column chromatography to obtain compound B15.

[0158] 4.12 mL of trifluoroacetic acid was added to a 500 mg solution of compound B15 in dichloromethane, and the mixture was stirred at room temperature for 1 h. The solvent was removed by vacuum evaporation, and the crude product containing C15 was directly used for the next reaction without purification.

[0159] 284.53 mg of the crude product of compound C15 was dissolved in 10 mL of tetrahydrofuran. The pH of the solution was adjusted to neutral by adding an appropriate amount of triethylamine. The reaction was carried out at room temperature for 30 min. Then, 1.98 g of compound D15 and NaBH(AcO)3 (10.76 mmol) were added. The reaction was carried out at room temperature for 24 h. The solvent was removed by vacuum distillation. The crude product was purified by silica gel column chromatography to obtain compound E15.

[0160] 433.08 mg of compound E15 was dissolved in a mixture of 5 mL dichloromethane and 5 mL DMF. 660.97 mg F15, 211.03 mg EDC and 8.3 mg DMAP were added to the mixture sequentially. The mixture was stirred overnight at room temperature. The solvent was removed by rotary evaporation under reduced pressure. The product was purified by silica gel column chromatography to obtain 1.056 g of the target compound G15, with a yield of 62%. The structure of the product was identified by HRMS, HRMS [M+H]+: 1964.8171.

[0161] Example 16

[0162]

[0163] The synthesis method of compound G16 is the same as in Example 1, and the structure of the product was identified by HRMS.

[0164] Example 17

[0165]

[0166] 4.51 g of the starting compound N-methyl-2,2-diaminodiethylamine was dissolved in 50 mL of dichloromethane. Under stirring, 20 mL of a dichloromethane solution containing 4.2 g of Boc₂O was added dropwise over 3 hours. After the reaction was complete, the organic phase was washed three times successively with 20 mL of 1 mol / L sodium bicarbonate aqueous solution and 20 mL of saturated brine, dried over anhydrous magnesium sulfate, and the solvent was removed by vacuum evaporation to obtain the target compound A17.

[0167] 2.29 g of compound A17 was dissolved in 10 mL of DMF solution, and 1.22 g of triethylamine and 5 mL of DMF solution containing 924 mg of bromoacetic acid were added with stirring. The reaction was carried out at room temperature for 24 h, and the solvent was removed by vacuum distillation. The crude product was purified by silica gel column chromatography to obtain compound B17.

[0168] 7.92 mL of trifluoroacetic acid was added to a dichloromethane solution of 1.04 g of compound B17, and the mixture was stirred at room temperature for 1 h. The solvent was removed by vacuum evaporation, and the crude product containing C17 was directly used for the next reaction without purification.

[0169] 641.56 mg of the crude product of compound C17 was dissolved in 10 mL of tetrahydrofuran solution, 805.45 mg of triethylamine was added, and the reaction was carried out at room temperature for 30 min. Then, 3.95 g of compound D17 and NaBH(OAc)3 (19.9 mmol) were added, and the reaction was carried out at room temperature for 24 h. The solvent was removed by vacuum distillation, and the crude product was purified by silica gel column chromatography to obtain compound E17.

[0170] 353.25 mg of compound E17 was dissolved in a mixture of 2 mL dichloromethane and 2 mL DMF. 198.95 mg F13, 73.9 mg EDC, and 2.91 mg DMAP were added to the mixture sequentially. The mixture was stirred overnight at room temperature. The solvent was removed by rotary evaporation under reduced pressure. The product was purified by silica gel column chromatography to obtain 238.01 mg of the target compound G17, with a yield of 43%. The structure of the product was identified by HRMS, HRMS [M+H]+: 2451.8107.

[0171] Example 18

[0172]

[0173] 1.54 g of the starting compound 1,3-propanediamine was dissolved in 20 mL of dichloromethane. Under stirring, 20 mL of a dichloromethane solution containing 2.27 g of Boc₂O was added dropwise over 3 hours. After the reaction was complete, the organic phase was washed three times successively with 20 mL of 1 mol / L sodium bicarbonate aqueous solution and 20 mL of saturated brine, dried over anhydrous magnesium sulfate, and the solvent was removed by vacuum evaporation to obtain the target compound A18.

[0174] 1.26 g of compound A18 was dissolved in 10 mL of DMF solution, and 733.87 mg of triethylamine and 5 mL of DMF solution containing 504 mg of 3-bromo-1-propanol were added with stirring. The reaction was carried out at room temperature for 24 h, and the solvent was removed by vacuum distillation. The crude product was purified by silica gel column chromatography to obtain compound B18.

[0175] 3.64 mL of trifluoroacetic acid was added to a 442 mg solution of compound B18 in dichloromethane, and the mixture was stirred at room temperature for 1 h. The solvent was removed by vacuum evaporation, and the crude product containing C18 was directly used for the next reaction without purification.

[0176] 500 mg of the crude product of compound C18 was dissolved in 10 mL of tetrahydrofuran. The pH of the solution was adjusted to neutral by adding an appropriate amount of triethylamine. The reaction was carried out at room temperature for 30 min. Then, 3.22 g of compound D18 and NaBH(AcO)3 (18.91 mmol) were added, and the reaction was carried out at room temperature for 24 h. The solvent was removed by vacuum distillation, and the crude product was purified by silica gel column chromatography to obtain compound E18.

[0177] 378.25 mg of compound E18 was dissolved in a mixture of 5 mL dichloromethane and 5 mL DMF. 379.26 mg of F18, 197.35 mg of EDC, and 7.77 mg of DMAP were added to the mixture sequentially. The mixture was stirred overnight at room temperature. The solvent was removed by rotary evaporation under reduced pressure. The product was purified by silica gel column chromatography to obtain 259.1 mg of the target compound G18, with a yield of 34%. The structure of the product was identified by HRMS, HRMS [M+H]+: 1172.8956.

[0178] Example 19

[0179]

[0180] 1.31 g of the starting compound was dissolved in 10 mL of dichloromethane. With stirring, 10 mL of a dichloromethane solution containing 1.88 g of Boc₂O was added dropwise over 3 hours. After the reaction was complete, the organic phase was washed three times successively with 20 mL of 1 mol / L sodium bicarbonate aqueous solution and 20 mL of saturated brine, dried over anhydrous magnesium sulfate, and the solvent was removed by vacuum evaporation to obtain the target compound A19.

[0181] 1.15 g of compound A19 was dissolved in 10 mL of DMF solution, and 661.5 mg of triethylamine and 5 mL of DMF solution containing 500 mg of bromoacetic acid were added with stirring. The reaction was carried out at room temperature for 24 h, and the solvent was removed by vacuum distillation. The crude product was purified by silica gel column chromatography to obtain compound B19.

[0182] 6.25 mL of trifluoroacetic acid was added to a dichloromethane solution of 811.5 mg of compound B19, and the mixture was stirred at room temperature for 1 h. The solvent was removed by vacuum evaporation, and the crude product containing C19 was directly used for the next reaction without purification.

[0183] 484.27 mg of the crude product of compound C19 was dissolved in 5 mL of tetrahydrofuran solution, 661.5 mg of triethylamine was added, and the reaction was carried out at room temperature for 30 min. Then, 3.93 g of compound D19 and NaBH(OAc)3 (16.34 mmol) were added, and the reaction was carried out at room temperature for 24 h. The solvent was removed by vacuum distillation, and the crude product was purified by silica gel column chromatography to obtain compound E19.

[0184] 327 mg of compound E19 was dissolved in a mixture of 2 mL dichloromethane and 2 mL DMF. 256.07 mg F19, 116.79 mg EDC and 4.60 mg DMAP were added to the mixture sequentially. The mixture was stirred overnight at room temperature. The solvent was removed by rotary evaporation under reduced pressure. The product was purified by silica gel column chromatography to obtain 176.15 mg of the target compound G19, with a yield of 30%. The structure of the product was identified by HRMS, HRMS [M+H]+: 1531.9467.

[0185] Example 20

[0186]

[0187] 2.23 g of the starting compound 1,3-propanediamine was dissolved in 20 mL of dichloromethane. Under stirring, 20 mL of a dichloromethane solution containing 3.29 g of Boc₂O was added dropwise over 3 hours. After the reaction was complete, the organic phase was washed three times successively with 20 mL of 1 mol / L sodium bicarbonate aqueous solution and 20 mL of saturated brine, dried over anhydrous magnesium sulfate, and the solvent was removed by vacuum evaporation to obtain the target compound A₂O.

[0188] 2.51 g of compound A20 was dissolved in 10 mL of DMF solution, and 1.46 g of triethylamine and 10 mL of DMF solution containing 1 g of 3-bromo-1-propanol were added with stirring. The mixture was reacted at room temperature for 24 h, and the solvent was removed by vacuum distillation. The crude product was purified by silica gel column chromatography to obtain compound B20.

[0189] 13.75 mL of trifluoroacetic acid was added to a 1.67 g solution of compound B18 in dichloromethane, and the mixture was stirred at room temperature for 1 h. The solvent was removed by vacuum evaporation, and the crude product containing C20 was directly used for the next reaction without purification.

[0190] 211.5 mg of the crude product of compound C20 was dissolved in 10 mL of tetrahydrofuran. The pH of the solution was adjusted to neutral by adding an appropriate amount of triethylamine. The reaction was carried out at room temperature for 30 min. Then, 2.12 g of compound D20 and NaBH(AcO)3 (8 mmol) were added, and the reaction was carried out at room temperature for 24 h. The solvent was removed by vacuum distillation, and the crude product was purified by silica gel column chromatography to obtain compound E20.

[0191] 323.77 mg of compound E20 was dissolved in a mixture of 5 mL dichloromethane and 5 mL DMF. 196.51 mg F18, 114.55 mg EDC and 4.51 mg DMAP were added to the mixture sequentially. The mixture was stirred overnight at room temperature. The solvent was removed by rotary evaporation under reduced pressure. The product was purified by silica gel column chromatography to obtain 178.94 mg of the target compound G20, with a yield of 34%. The structure of the product was identified by HRMS, HRMS [M+H]+: 1392.1177.

[0192] Example 21

[0193]

[0194] 2 g of the starting compound hydroxyethyl ethylenediamine (19.2 mmol) was dissolved in 50 mL of tetrahydrofuran. 27.31 g of compound A21 and NaBH(AcO)3 (96.01 mmol) were added with stirring. The mixture was reacted at room temperature for 24 h. The solvent was removed by vacuum distillation. The crude product was purified by silica gel column chromatography to obtain compound B21.

[0195] 885.31 mg of compound B21 was dissolved in 10 mL of dry CH2Cl2, followed by the addition of 197.01 mg of triethylamine and a catalytic amount of DMAP. The mixture was then added dropwise with stirring at 0 °C in 10 mL of dichloromethane containing 278.36 mg of p-toluenesulfonyl chloride. After the reaction was complete, the solvent was removed by vacuum distillation, and the mixture was redissolved in 10 mL of DMF, followed by the addition of 126.57 mg of NaN3. The reaction was carried out at 80 °C for 5 h. After the reaction was complete, the organic phase was washed repeatedly with small amounts of 20 mL of saturated brine, dried over anhydrous sodium sulfate, and the solvent was removed by rotary evaporation under reduced pressure. The crude product was purified by silica gel column chromatography to obtain compound C21.

[0196] Under argon protection, 509.66 mg of compound C21 was dissolved in 20 mL of ultra-dry tetrahydrofuran, and 62.1 mg of lithium aluminum hydride was added at 0 °C. The reaction was stirred at room temperature for 30 min, and quenched by the slow addition of 5 mL of water and 5 mL of 15% sodium hydroxide aqueous solution. The organic phase was washed repeatedly with 20 mL of saturated brine, dried over anhydrous sodium sulfate, and the solvent was removed under reduced pressure. The crude product was purified by silica gel column chromatography to obtain compound D21.

[0197] 76.39 mg of compound E21 was dissolved in a mixed solution of 3 mL CH2Cl2 and 1 mL MeCN. Then, 159.86 mg of 2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate and 42.54 mg of triethylamine were added sequentially to the mixed solution. The mixture was reacted at 0 °C in an ice bath for 30 min, followed by the addition of 25.17 mg of compound D21. The reaction was stirred overnight at room temperature. The solvent was removed by rotary evaporation under reduced pressure. The product was purified by silica gel column chromatography to obtain 42.87 mg of the target compound G21, with a yield of 42%. The product structure was identified by HRMS [M+H]. + :2378.9246.

[0198] Example 22

[0199]

[0200] 1.3 g of the starting compound 1,4-cyclohexylamine dimethylamine was dissolved in 20 mL of dichloromethane. Under stirring, 20 mL of a dichloromethane solution containing 1 g of Boc₂O was added dropwise over 3 hours. After the reaction was complete, the organic phase was washed three times successively with 20 mL of 1 mol / L sodium bicarbonate aqueous solution and 20 mL of saturated brine, dried over anhydrous sodium sulfate, and the solvent was removed by vacuum evaporation to obtain the target compound A22.

[0201] 899.76 mg of compound A22 was dissolved in 10 mL of DMF solution, and 375.67 mg of triethylamine and 5 mL of DMF solution containing 258 mg of 3-bromo-1-propanol were added with stirring. The reaction was carried out at room temperature for 24 h, and the solvent was removed by vacuum distillation. The crude product was purified by silica gel column chromatography to obtain compound B22.

[0202] 3.55 mL of trifluoroacetic acid was added to a dichloromethane solution containing 557.69 mg of compound B22, and the mixture was stirred at room temperature for 1 h. The solvent was removed by vacuum evaporation, and the crude product containing C22 was directly used for the next reaction without purification.

[0203] 217.85 mg of the crude product of compound C22 was dissolved in 10 mL of tetrahydrofuran, and an appropriate amount of triethylamine was added to adjust the pH of the solution to neutral. The reaction was carried out at room temperature for 30 min, and then 773.42 mg of compound D22 and NaBH(AcO)3 (5.44 mmol) were added. The reaction was carried out at room temperature for 24 h, and the solvent was removed by vacuum evaporation. The crude product was purified by silica gel column chromatography to obtain compound E22.

[0204] 47.4 mg of compound E22 was dissolved in a mixture of 2 mL dichloromethane and 2 mL DMF. 23.03 mg F22, 12.71 mg EDC and 0.5 mg DMAP were added to the mixture sequentially. The mixture was stirred overnight at room temperature. The solvent was removed by rotary evaporation under reduced pressure. The product was purified by silica gel column chromatography to obtain 42.37 mg of the target compound G22, with a yield of 61%. The structure of the product was identified by HRMS, HRMS [M+H]+: 1684.4631.

[0205] Example 23

[0206]

[0207] 3.1 g of the starting compound ethylenediamine was dissolved in 20 mL of dichloromethane. Under stirring, 20 mL of a dichloromethane solution containing 5.62 g of Boc₂O was added dropwise over 3 hours. After the reaction was complete, the organic phase was washed three times successively with 20 mL of 1 mol / L sodium bicarbonate aqueous solution and 20 mL of saturated brine, dried over anhydrous magnesium sulfate, and the solvent was removed by vacuum evaporation to obtain the target compound A23.

[0208] 337.66 mg of compound A23 was dissolved in 10 mL of DMF solution, and 213.27 mg of triethylamine and 10 mL of DMF solution containing 161.2 mg of bromoacetic acid were added with stirring. The reaction was carried out at room temperature for 24 h, and the solvent was removed by vacuum distillation. The crude product was purified by silica gel column chromatography to obtain compound B23.

[0209] 2.02 mL of trifluoroacetic acid was added to a dichloromethane solution containing 244.77 mg of compound B23, and the mixture was stirred at room temperature for 1 h. The solvent was removed by vacuum evaporation, and the crude product containing C23 was directly used for the next reaction without purification.

[0210] 139.27 mg of the crude product of compound C23 was dissolved in 10 mL of tetrahydrofuran solution, 213.27 mg of triethylamine was added, and the reaction was carried out at room temperature for 30 min. Then, 1.05 g of compound D23 and NaBH(OAc)3 (5.27 mmol) were added, and the reaction was carried out at room temperature for 24 h. The solvent was removed by vacuum distillation, and the crude product was purified by silica gel column chromatography to obtain compound E23.

[0211] 21.97 mg of compound E23 was dissolved in a mixed solution of 3 mL CH2Cl2 and 1 mL MeCN. 121.93 mg of 2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate and 32.45 mg of triethylamine were added sequentially to the mixed solution. The mixture was reacted at 0 °C in an ice bath for 30 min, followed by the addition of 20.88 mg of compound D23. The reaction was stirred overnight at room temperature. The solvent was removed by rotary evaporation under reduced pressure. The product was purified by silica gel column chromatography to obtain 18.27 mg of the target compound G23, with a yield of 43%. The product structure was identified by HRMS [M+H]. + :1315.7592.

[0212] Example 24

[0213]

[0214] 6.13 g of the starting compound 1,3-propanediamine was dissolved in 50 mL of dichloromethane. Under stirring, 50 mL of a dichloromethane solution containing 9.02 g of Boc₂O was added dropwise over 3 hours. After the reaction was complete, the organic phase was washed three times successively with 30 mL of 1 mol / L sodium bicarbonate aqueous solution and 30 mL of saturated brine, dried over anhydrous magnesium sulfate, and the solvent was removed by vacuum evaporation to obtain the target compound A24.

[0215] 6.52 g of compound A24 was dissolved in 20 mL of DMF solution, and 3.78 g of triethylamine and 20 mL of DMF solution containing 2.86 g of bromoacetic acid were added with stirring. The reaction was carried out at room temperature for 24 h, and the solvent was removed by vacuum distillation. The crude product was purified by silica gel column chromatography to obtain compound B24.

[0216] 3.57 mL of trifluoroacetic acid was added to a 460 mg solution of compound B24 in dichloromethane, and the mixture was stirred at room temperature for 1 h. The solvent was removed by vacuum evaporation, and the crude product containing C24 was directly used for the next reaction without purification.

[0217] 273.02 mg of the crude product of compound C24 was dissolved in 10 mL of tetrahydrofuran. The pH of the solution was adjusted to neutral by adding an appropriate amount of triethylamine. The reaction was carried out at room temperature for 30 min. Then, 1.48 g of compound D24 and NaBH(AcO)3 (9.34 mmol) were added. The reaction was carried out at room temperature for 24 h. The solvent was removed by vacuum distillation. The crude product was purified by silica gel column chromatography to obtain compound E24.

[0218] 100 mg of compound E24 was dissolved in a mixture of 2 mL dichloromethane and 2 mL DMF. 94.89 mg F24, 54.21 mg EDC and 2.13 mg DMAP were added to the mixture sequentially. The mixture was stirred overnight at room temperature. The solvent was removed by rotary evaporation under reduced pressure. The product was purified by silica gel column chromatography to obtain 104.59 mg of the target compound G24, with a yield of 54%. The structure of the product was identified by HRMS, HRMS [M+H]+: 1139.5672.

[0219] Example 24

[0220] The preparation methods of compounds G25-G30 are the same as those described in the examples above.

[0221]

[0222]

[0223]

[0224] Example 26

[0225] Cytotoxicity assays of high concentrations of compound G1:

[0226] An ethanol phase was prepared by mixing 2 mg / mL G1 lipid molecules, 2 mg / mL phospholipids, 2 mg / mL cholesterol, and 0.4 mg / mL polyethylene glycol in a molar ratio of 25:50:50:3, and thoroughly mixing. Calf thymus DNA (Ct-DNA) was dissolved in 10 mmol / L pH 3 sodium citrate buffer to prepare a DNA premix with a DNA concentration of 0.004 μg / μL. The DNA premix (volume ratio 1:1) was added to the ethanol phase and thoroughly mixed. The mixture was then diluted with an equal volume of phosphate buffer to obtain the G1 lipid nanoparticle stock solution.

[0227] 24 hours before treatment, HeLa cells were injected at a rate of 4 × 10⁻⁶. 3 Cells were cultured at a density of 100 μL / well in 96-well plates, with each well containing 10% (v / v) FBS and 1% penicillin / streptomycin. The G1 lipid nanoparticle stock solution was diluted with MEM medium to obtain a lipid nanoparticle solution with an equivalent photosensitizer concentration of 5 μmol / L. The original medium was removed, and 100 μL of the lipid nanoparticle solution was added to each well, with four auxiliary wells for each experimental group.

[0228] After incubating the cells for 18 hours, at 808 nm, W = 700 mW / cm 2 Cells were exposed to light for 15 minutes, the culture medium was removed, and 120 μL of a mixture of culture medium and MTT (culture medium to MTT volume ratio of 100:20, initial MTT stock solution concentration of 5 mg / mL) was added to each well. Cells were incubated at 37 °C for 4 hours. The culture medium was then removed, and 150 μL of dimethyl sulfoxide was added. After shaking for 10 minutes, the absorbance was measured at 490 nm using a SpectraMax M2e microplate reader. Cell viability of G1-treated cells was normalized to that of untreated cells.

[0229] Cytotoxicity results such as Figure 1 As shown, cells were incubated with a high concentration of G1 for 18 hours at 808 nm and W = 300 mW / cm². 2 The cell survival rate under light conditions showed a statistically significant difference compared to that under dark conditions, indicating that after G1 lipid nanoparticles are taken up by cells, they can release photosensitizers through hydrolysis, and can generate ROS to induce cell death under light.

[0230] Example 27

[0231] Cytotoxicity assays of high concentrations of compound G15:

[0232] An ethanol phase was prepared by mixing 2 mg / mL G15 lipid molecules, 2 mg / mL phospholipids, 2 mg / mL cholesterol, and 0.4 mg / mL polyethylene glycol in an ethanol solution at a molar ratio of 20:30:40:0.75 and thoroughly mixing. Calf thymus DNA (Ct-DNA) was dissolved in 10 mmol / L pH 3 sodium citrate buffer to prepare a DNA premix with a DNA concentration of 0.004 μg / μL. The DNA premix (volume ratio 1:1) was added to the ethanol phase and thoroughly mixed. The mixture was then diluted with an equal volume of phosphate buffer to obtain the G15 lipid nanoparticle stock solution.

[0233] 24 hours before treatment, 4T1 cells were loaded at 4 × 10⁻⁶. 3 Cells were cultured at a density of 100 μL per well in 96-well plates, with each well containing 10% (v / v) FBS and 1% penicillin / streptomycin. The G15 lipid nanoparticle stock solution was diluted with DMEM to obtain a lipid nanoparticle solution with an equivalent photosensitizer concentration of 5 μmol / L. The original culture medium was removed, and 100 μL of the lipid nanoparticle solution was added to each well, with four auxiliary wells for each experimental group.

[0234] After incubating the cells for 18 hours, at 640 nm, W=50 mW / cm2 Cells were exposed to light for 15 minutes, the culture medium was removed, and 120 μL of a mixture of culture medium and MTT (culture medium to MTT volume ratio of 100:20, initial MTT stock solution concentration of 5 mg / mL) was added to each well. Cells were incubated at 37 °C for 4 hours. The culture medium was then removed, and 150 μL of dimethyl sulfoxide was added. After shaking for 10 minutes, the absorbance was measured at 490 nm using a SpectraMax M2e microplate reader. Cell viability of G15-treated cells was normalized to that of untreated cells.

[0235] Cytotoxicity results such as Figure 2 As shown, cells were incubated with a high concentration of G15 for 18 hours at 640 nm and W = 50 mW / cm². 2 The cell survival rate under light conditions showed a statistically significant difference compared to that under dark conditions, indicating that after G15 lipid nanoparticles are taken up by cells, they can release photosensitizers through hydrolysis, and can generate ROS to induce cell death under light.

[0236] Example 28

[0237] Tests on the ability of lipid molecule G1 to release photosensitizers and induce intracellular ROS production:

[0238] The preparation process of the mother liquor for G1 type lipid nanoparticles is as described in Example 26.

[0239] 24 hours before treatment, HeLa cells were injected at 8 × 10⁻⁶. 4 Cells were cultured at a density of 1 mL in confocal culture dishes containing 10% (v / v) FBS and 1% penicillin / streptomycin. After incubating cells with G1 lipid nanoparticles at a photosensitizer equivalent concentration of 2.5 μmol / L for 18 hours, the culture medium was removed, and the cells were washed three times with phosphate buffer. Cells were then incubated with the ROS detection probe DCFH-DA at a concentration of 10 μmol / L for 20 minutes, the culture medium was removed, and the cells were washed three times with serum-free MEM medium. Cells were cultured at 808 nm with a W = 300 mW / cm². 2 Irradiated for 15 minutes under the specified conditions. Imaging was performed using a single-photon laser confocal microscope (Olympus FV1000), with the microscope excitation wavelength set to 488 nm and the detector detection wavelength range to 500–550 nm.

[0240] Experimental results are as follows Figure 3As shown, intracellular singlet oxygen can oxidize non-fluorescent DCFH to fluorescent DCF. Results showed no significant fluorescence signal in the control group and the dark treatment group with added G1 lipid nanoparticles; however, a significant fluorescence signal was observed in the light treatment group with added G1 lipid nanoparticles. In summary, G1 can induce ROS production in cells under light conditions through the release of photosensitizers, producing the same effect as photosensitizers.

[0241] Example 29

[0242] Tests on the ability of lipid molecule G15 to release photosensitizers and induce intracellular ROS production:

[0243] The preparation process of the G15 lipid nanoparticle mother liquor is as described in Example 27.

[0244] 24 hours before treatment, 4T1 cells were loaded at 8 × 10⁻⁶. 4 Cells were cultured at a density of 1 mL in confocal culture dishes containing 10% (v / v) FBS and 1% penicillin / streptomycin. After incubating the cells with G1 lipid nanoparticles at a photosensitizer equivalent concentration of 5 μmol / L for 18 hours, the culture medium was removed, and the cells were washed three times with phosphate buffer. The ROS detection probe DCFH-DA at a concentration of 10 μmol / L was added to the confocal dish, and the cells were incubated for 20 minutes. After removing the culture medium, the cells were washed three times with serum-free DMEM and cultured at 640 nm, W = 50 mW / cm². 2 Irradiated for 15 minutes under the specified conditions. Imaging was performed using a single-photon laser confocal microscope (Olympus FV1000), with the microscope excitation wavelength set to 488 nm and the detector detection wavelength range to 500–550 nm.

[0245] Experimental results are as follows Figure 4 As shown, intracellularly generated singlet oxygen can oxidize non-fluorescent DCFH to fluorescent DCF. Results showed no significant fluorescence signal in the control group and the dark treatment group with added G15 lipid nanoparticles; however, a significant fluorescence signal was observed in the light treatment group with added G15 lipid nanoparticles. In summary, G15 can induce ROS production in cells under light conditions through photosensitizer release, producing the same effect as photosensitizers.

[0246] Example 30

[0247] G1 cell staining experiment with lipid molecules:

[0248] The preparation process of the mother liquor for G1 type lipid nanoparticles is as described in Example 26.

[0249] 24 hours before treatment, 4T1 cells were loaded at 8 × 10⁻⁶. 4 Cells were cultured at a density of 1 mL in confocal culture dishes containing 10% (v / v) FBS and 1% penicillin / streptomycin. After incubation for 18 hours with G1 lipid nanoparticles at a photosensitizer equivalent concentration of 10 μmol / L, the culture medium was removed, and the cells were washed three times with phosphate buffer solution. Cells were then cultured at 808 nm (W = 100 mW / cm²). 2 Cells were irradiated for 15 minutes under the specified conditions, and then incubated in a confocal dish with 8 μmol / L Calcein AM and 2 μmol / L PI working solution for 20 minutes. Imaging was performed using a single-photon laser confocal microscope (Olympus FV1000). The excitation wavelength of Channel 1 was set to 488 nm, and the detector detection wavelength range was 500–550 nm. The excitation wavelength of Channel 2 was set to 559 nm, and the detector detection wavelength range was 600–700 nm.

[0250] Experimental results are as follows Figure 5 As shown, live cells exhibit fluorescence signals in Channel 1, while dead cells exhibit fluorescence signals in Channel 2. The results showed that no fluorescence signal was observed in Channel 2 in the control group and the G1 dark treatment group; however, Channel 2 showed a certain fluorescence intensity in the G1 light-treated group. In summary, G1 can induce ROS production in cells under light conditions through the release of photosensitizers, which can induce damage and necrosis of tumor cells.

[0251] Example 31

[0252] G15 cell staining experiment for lipid molecules:

[0253] The preparation process of the G15 lipid nanoparticle mother liquor is as described in Example 27.

[0254] 24 hours before treatment, 4T1 cells were loaded at 8 × 10⁻⁶. 4 Cells were cultured at a density of 1 mL in confocal culture dishes containing 10% (v / v) FBS and 1% penicillin / streptomycin. After incubation for 18 hours with G1 lipid nanoparticles at a photosensitizer equivalent concentration of 5 μmol / L, the culture medium was removed, and the cells were washed three times with phosphate buffer solution. Cells were then cultured at 649 nm (W = 50 mW / cm²). 2Cells were irradiated for 15 minutes under the specified conditions, and then incubated in a confocal dish with 8 μmol / L Calcein AM and 2 μmol / L PI working solution for 20 minutes. Imaging was performed using a single-photon laser confocal microscope (Olympus FV1000). The excitation wavelength of Channel 1 was set to 488 nm, and the detector detection wavelength range was 500–550 nm. The excitation wavelength of Channel 2 was set to 559 nm, and the detector detection wavelength range was 600–700 nm.

[0255] Experimental results are as follows Figure 6 As shown, live cells exhibit fluorescence signals in Channel 1, while dead cells exhibit fluorescence signals in Channel 2. The results showed that no fluorescence signal was observed in Channel 2 in the control group and the G15 dark treatment group; however, Channel 2 showed a certain fluorescence intensity in the G15 light-treated group. In summary, G15 can induce ROS production in cells under light conditions through the release of photosensitizers, which can induce damage and necrosis of tumor cells.

[0256] Example 32

[0257] Tests of G1's ability to deliver eGFP mRNA to live cells using lipid nanoparticles:

[0258] An ethanol phase was prepared by mixing 2 mg / mL G1 lipid molecules, 2 mg / mL phospholipids, 2 mg / mL cholesterol, and 0.4 mg / mL polyethylene glycol in a molar ratio of 20:30:40:7.5 and thoroughly mixing. Green fluorescent protein messenger RNA (eGFP mRNA) was diluted with 10 mmol / L pH=3 sodium citrate buffer to prepare an mRNA premix with a concentration of 0.004 μg / μL. The mRNA premix was added to the ethanol phase (V / V = 1:1) and thoroughly mixed. The above solution was then diluted with an equal volume of phosphate-buffered saline (PBS) to obtain the G1 lipid nanoparticle stock solution.

[0259] 24 hours before treatment, HeLa cells were injected at 8 × 10⁻⁶. 4 The samples were cultured at a density in confocal culture dishes, each containing 1 mL of MEM medium. This medium contained 10% (v / v) FBS and 1% penicillin / streptomycin.

[0260] 250 μL of G1 lipid nanoparticle stock solution was added to 2 ml of culture medium and thoroughly mixed, then the cells were incubated for 18 h. After 18 h, the culture medium was removed, and the cells were washed three times with phosphate buffer. The expression of green fluorescent protein was detected using a single-photon laser confocal microscope (Olympus FV1000), with the microscope excitation wavelength set at 488 nm and the detector detection wavelength at 500–550 nm.

[0261] Experimental results are as follows Figure 7 As shown, the experimental results indicate that lipid nanoparticles based on compound G1 can deliver eGFP mRNA to living cells, and some cells show obvious green fluorescence, indicating successful expression of EGFP protein.

[0262] Example 33

[0263] The cytotoxicity experiments and tests of the ability of compounds G3, G4, G7, G8, G9, G10, G11, G12, G13, G14, G17, G18, and G19 to induce intracellular ROS production were conducted according to the methods in Examples 26 and 28. The experiments showed that compounds G3, G4, G7, G8, G9, G10, G11, G12, G13, G14, G17, G18, and G19 can all be taken up by cells and release photosensitizers through hydrolysis. The release of photosensitizers induces the production of ROS in cells under light conditions, producing the same effect as photosensitizers.

Claims

1. Compounds of formula I or their salts: in, R0 is independent of each , , ; R1 is independent of each other. , , , , , , or ; n are independent integers from 1 to 5; R2 is independent of each other. , , C6-C 20 Saturated alkane group or C6-C 20 The unsaturated alkane groups, wherein each R3 is independently C4-C6. 16 saturated alkane group, C4-C 16 unsaturated alkane groups or R4 is independent of each other. , or Each of p is an independent integer from 1 to 18; m is 1 or 2; X is a carbonyl group linked to a photosensitizer; The photosensitizer is selected from phthalocyanine, cyanine dyes, fluorescein, or doxorubicin.

2. The compound or its salt according to claim 1, characterized in that: X is independent of each other. , , , , or ; Among them, Y - Each anion is independent and selected from BF4. - Cl - ,Br - I - NO 3- SO4 2- ClO4 - CH3COO - CH3SO3 - CF3SO3 - .

3. The compound or its salt according to claim 2, characterized in that: R1 is independent of each other. , , , , , , , , or .

4. The compound or its salt according to claim 3, characterized in that: R2 is independent of each other. , , C8-C 18 Saturated alkane group or C8-C 18 The unsaturated alkane groups, wherein each R3 is independently C6-C6. 14 saturated alkane group, C6-C 14 unsaturated alkane groups or R4 is independent of each other. , or p are independent integers from 0 to 12.

5. The compound or its salt according to claim 4, characterized in that: R0 is independent of each , , or .

6. The compound or its salt according to claim 4, characterized in that: R2 is independent of each other. , , , , , , , , , , , , , , , , , , , , , , , , , , , or ; Among them, R3 is independent of each other. , , , , , , , , , , , , , , , , , , , , , , or ; R4 is independent of each , or .

7. The compound or a salt thereof according to claim 6, characterized in that: R2 is independent of each other. , , , , , , , , , , , , , , , , , , , , or ; Among them, R3 is independent of each other. , , , , , , , , , , , , , , , or ; R4 is independent of each , or .

8. A pharmaceutical composition, characterized in that, Includes the compound or salt thereof as described in any one of claims 1-7.

9. A nanomaterial, characterized in that, Includes the compound or salt thereof as described in any one of claims 1-7.

10. The nanomaterial according to claim 9, characterized in that, It also includes RNA or DNA.

11. Use of the compound according to any one of claims 1-7 in the preparation of photosensitizing drugs or materials for delivering RNA / DNA.

12. The use of the compound according to any one of claims 1-7 in the preparation of a dual-mode synergistic therapeutic agent for gene therapy and photodynamic therapy.

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