5'-amino-2', 5'-dideoxynucleotide phospholipid molecule and its preparation method and application

Abstract

The invention discloses 5'-amino-2',5'-dideoxynucleotide phospholipid molecules, and a preparation method and an application thereof. The 5'-amino-2',5'-dideoxynucleotide phospholipid molecules provided by the invention have an amphiphilic structure represented by the formula (I). 5'-amino-2',5'-dideoxynucleoside is adopted as a hydrophilic head, and glycerol fatty ether connected by using a phosphate bond is adopted as a hydrophobic tail, such that an amphiphilic molecule is formed. The Base group is common purine and pyrimidine bases, and R group is saturated or unsaturated aliphatic carbon chain. In a water-phase solvent, the dideoxynucleotide phospholipid molecule can form a plurality of supramolecular structures including multi-layer film structure, lipid structure, fibrous structure, and hydrogel. Therefore, 5'-amino-2',5'-dideoxynucleotide phospholipid molecules have certain potential to be used as aggregation, trapping, loading, or carrying type nucleic acid medicines, and can be used as a high-efficiency biological material for mediating transmembrane of nucleic acid medicines.

Description

technical field

[0001] The present invention relates to a series of 5'-amino-2', 5'-dideoxynucleotide phospholipid molecules and their chemical synthesis method, and also relates to this series of 5'-amino-2', 5'-dideoxynucleotide A supramolecular structure formed by the self-assembly of phospholipid molecules in an aqueous solvent. It belongs to the field of new biomaterials. Background technique

[0002] Introducing double-stranded RNA (siRNA, small interfering RNA) composed of sense RNA and antisense RNA corresponding to mRNA into cells can induce specific degradation of mRNA, resulting in corresponding gene silencing. This sequence-specific post-transcriptional gene silencing mechanism is called RNA interference (RNAi). As a drug with great potential, siRNA will open up a new way for the treatment of human diseases. However, there are many problems in the delivery of siRNA in vivo. For example, its serum stability is not good, and it is easily degraded by RNase in th...

Examples

Embodiment 1

[0066] [Example 1] Synthesis of 5'-toluenesulfonyl ester thymidine

[0067] Thymidine (24.2 g, 0.1 mol) was dissolved in anhydrous pyridine (200 mL), and cooled to 0° C. in an ice bath. p-Toluenesulfonyl chloride (23 g, 0.12 mol) was dissolved in anhydrous pyridine (100 mL), and slowly added dropwise to the aforementioned reaction solution through a microsampler, and the dropping process lasted for 6 hours. The reaction process was always carried out at 0°C under the protection of argon. After the dropwise addition was completed, the reaction was returned to room temperature, and stirring was continued for 6 h. Pyridine was distilled off under reduced pressure. To the residue was added ethyl acetate 500 mL and NaHCO 3 Aqueous solution (10%) 300mL, a large amount of white solid precipitated. After filtration, the obtained white solid is the target product. The filtrate was collected, the organic phase was separated, and the aqueous phase was washed with 300 mL of ethyl ace...

Embodiment 2

[0068] [Example 2] Synthesis of 5'-azido-2',5'-dideoxythymidine

[0069] Dissolve 5'-p-methylsulfonylcarbothymidine (5g, 12.6mmol) in anhydrous DMF (25mL), add NaN 3 (1.2g, 19mmol), heated to 70°C, and reacted for 12h under the protection of argon. The solvent was evaporated under reduced pressure. Dichloromethane (50 mL) was added to the residue, followed by washing with water (30 mL). Anhydrous Na for organic phase 2 SO 4 Dry and separate by silica gel column chromatography, eluent DCM:MeOH=20:1, to obtain 2.9 g of the target product (yield 85%). white solid. 1 HNMR (400MHz, DMSO-d 6):δ=11.31(s,1H),7.49(s,1H),6.20(t,J=7.2Hz,1H),5.39(d,J=4.0Hz,1H),4.20(s,1H),3.79 -3.85(m,1H),3.56(d,J=5.2Hz,2H),2.20-2.30(m,1H),2.05-2.15(m,1H),1.79(s,3H); 13 CNMR (100MHz, DMSO-d 6 ):δ=163.7, 150.5, 136.1, 109.8, 84.6, 83.9, 70.7, 51.6, 38.1, 12.1; 1298.8, 1272.8, 1067.3, 963.4, 856.2, 636.4, 553.4, 493.8cm -1 ;MS(ESI-TOF+)forC 10 h 13 N 5 o 4 Na[M+Na] + found290.1042,calcd290.08...

Embodiment 3

[0070] [Example 3] Synthesis of 5'-amino-2',5'-dideoxythymidine

[0071] Take 5'-azido-2',5'-dideoxythymidine (5.5g, 20mmol) and dissolve it in methanol (150mL), add 10% palladium carbon catalyst (0.55g), and place it in a hydrogenation apparatus hydrogenation. The pressure is 60Psi. After hydrogenation at room temperature for 5 h, the reaction was stopped. TLC detection found that the raw material had reacted completely. Suction filtration through celite under reduced pressure to remove the solid matter, and the obtained filtrate was evaporated to dryness to obtain 4.8 g of light yellow solid. The solid is insoluble in organic solvents such as methanol and ethyl acetate, and is very polar, and cannot be separated by ordinary silica gel column chromatography. The resulting solid was dissolved in water and purified by strongly acidic ion exchange resin Dowex50 to obtain 4.7 g of the target product with a yield of 95%. white solid. 1 HNMR (400MHz, DMSO-d 6 ):δ=7.65(s,1H),...

technical field

[0001] The present invention relates to a series of 5'-amino-2', 5'-dideoxynucleotide phospholipid molecules and their chemical synthesis method, and also relates to this series of 5'-amino-2', 5'-dideoxynucleotide A supramolecular structure formed by the self-assembly of phospholipid molecules in an aqueous solvent. It belongs to the field of new biomaterials. Background technique

[0002] Introducing double-stranded RNA (siRNA, small interfering RNA) composed of sense RNA and antisense RNA corresponding to mRNA into cells can induce specific degradation of mRNA, resulting in corresponding gene silencing. This sequence-specific post-transcriptional gene silencing mechanism is called RNA interference (RNAi). As a drug with great potential, siRNA will open up a new way for the treatment of human diseases. However, there are many problems in the delivery of siRNA in vivo. For example, its serum stability is not good, and it is easily degraded by RNase in th...

Examples

Embodiment 1

[0066] [Example 1] Synthesis of 5'-toluenesulfonyl ester thymidine

[0067] Thymidine (24.2 g, 0.1 mol) was dissolved in anhydrous pyridine (200 mL), and cooled to 0° C. in an ice bath. p-Toluenesulfonyl chloride (23 g, 0.12 mol) was dissolved in anhydrous pyridine (100 mL), and slowly added dropwise to the aforementioned reaction solution through a microsampler, and the dropping process lasted for 6 hours. The reaction process was always carried out at 0°C under the protection of argon. After the dropwise addition was completed, the reaction was returned to room temperature, and stirring was continued for 6 h. Pyridine was distilled off under reduced pressure. To the residue was added ethyl acetate 500 mL and NaHCO 3 Aqueous solution (10%) 300mL, a large amount of white solid precipitated. After filtration, the obtained white solid is the target product. The filtrate was collected, the organic phase was separated, and the aqueous phase was washed with 300 mL of ethyl ace...

Embodiment 2

[0068] [Example 2] Synthesis of 5'-azido-2',5'-dideoxythymidine

[0069] Dissolve 5'-p-methylsulfonylcarbothymidine (5g, 12.6mmol) in anhydrous DMF (25mL), add NaN 3 (1.2g, 19mmol), heated to 70°C, and reacted for 12h under the protection of argon. The solvent was evaporated under reduced pressure. Dichloromethane (50 mL) was added to the residue, followed by washing with water (30 mL). Anhydrous Na for organic phase 2 SO 4 Dry and separate by silica gel column chromatography, eluent DCM:MeOH=20:1, to obtain 2.9 g of the target product (yield 85%). white solid. 1 HNMR (400MHz, DMSO-d 6):δ=11.31(s,1H),7.49(s,1H),6.20(t,J=7.2Hz,1H),5.39(d,J=4.0Hz,1H),4.20(s,1H),3.79 -3.85(m,1H),3.56(d,J=5.2Hz,2H),2.20-2.30(m,1H),2.05-2.15(m,1H),1.79(s,3H); 13 CNMR (100MHz, DMSO-d 6 ):δ=163.7, 150.5, 136.1, 109.8, 84.6, 83.9, 70.7, 51.6, 38.1, 12.1; 1298.8, 1272.8, 1067.3, 963.4, 856.2, 636.4, 553.4, 493.8cm -1 ;MS(ESI-TOF+)forC 10 h 13 N 5 o 4 Na[M+Na] + found290.1042,calcd290.08...

Embodiment 3

[0070] [Example 3] Synthesis of 5'-amino-2',5'-dideoxythymidine

[0071] Take 5'-azido-2',5'-dideoxythymidine (5.5g, 20mmol) and dissolve it in methanol (150mL), add 10% palladium carbon catalyst (0.55g), and place it in a hydrogenation apparatus hydrogenation. The pressure is 60Psi. After hydrogenation at room temperature for 5 h, the reaction was stopped. TLC detection found that the raw material had reacted completely. Suction filtration through celite under reduced pressure to remove the solid matter, and the obtained filtrate was evaporated to dryness to obtain 4.8 g of light yellow solid. The solid is insoluble in organic solvents such as methanol and ethyl acetate, and is very polar, and cannot be separated by ordinary silica gel column chromatography. The resulting solid was dissolved in water and purified by strongly acidic ion exchange resin Dowex50 to obtain 4.7 g of the target product with a yield of 95%. white solid. 1 HNMR (400MHz, DMSO-d 6 ):δ=7.65(s,1H),...

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