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2'-C-methyl guanosine phosphoramidite monomer and synthesis method thereof

A technique for the synthesis of methylguanosine phosphoramidites and methods, which is applied in the fields of chemical instruments and methods, biochemical equipment and methods, and the preparation of sugar derivatives. Modify RNA technology development and other issues to achieve the effect of simple operation, low cost and easy cost

Inactive Publication Date: 2018-10-16
南京百芙利科技有限责任公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the high difficulty and cost of chemical synthesis limit the development of chemically modified RNA technology, and more and more complex phosphoramidite monomers need to be developed

Method used

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  • 2'-C-methyl guanosine phosphoramidite monomer and synthesis method thereof
  • 2'-C-methyl guanosine phosphoramidite monomer and synthesis method thereof
  • 2'-C-methyl guanosine phosphoramidite monomer and synthesis method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] N 2 - Synthesis of isobutyryl-2'-C-methylguanosine:

[0035]

[0036]In a 250ml two-necked bottle, add 75ml pyridine and 3.0g (10mmol) 2'-C-methylguanosine (self-made and commercialized) respectively, and slowly drop in 8.1g (75mmol) trimethyl chloride under the condition of nitrogen protection Silane, then stirred at 25°C for 2 hours. Under nitrogen protection and 0 DEG C condition, slowly dropwise add 2.1g (20mmol) isobutyryl chloride, then 25 DEG C of stirring reaction 3 hours. Cool to 0°C, add dropwise 10ml of water, then stir the reaction at 25°C for 20 minutes, add dropwise 25ml of saturated NH 4 OH aqueous solution, then stirred the reaction at 25°C for 30 minutes. Add 150ml of water and 100ml of dichloromethane, stir for 10 minutes, let stand for liquid separation, spin dry the solvent, and then recrystallize with 5ml of water to obtain 3.0g N 2 -Isobutyryl-2'-C-methylguanosine, yield 82%.

Embodiment 2

[0038] 5'-O-(4,4-Dimethoxytrityl)-N 2 - Synthesis of isobutyryl-2'-C-methylguanosine:

[0039]

[0040] In a 250ml single-necked bottle, add 1.8g (5mmol) N 2 -Isobutyryl-2'-C-methylguanosine, after azeotropic removal of water with pyridine, add 50ml of pyridine, then slowly add 1.7g (5mmol) 4,4'-bismethoxytrityl chloride dropwise 10ml of pyridine solution, then stirred at 25°C for 5 hours. Then slowly add 50ml of 5% sodium bicarbonate aqueous solution dropwise, extract three times with ethyl acetate (50ml x 3), dry with anhydrous sodium sulfate, spin the solvent, and then use 20ml (ethyl acetate / petroleum ether=3:1) Recrystallization gave 2.5 g of 5'-O-(4,4-dimethoxytrityl)-N 2 -Isobutyryl-2'-C-methylguanosine, yield 75%.

Embodiment 3

[0042] 5'-O-(4,4-dimethoxytrityl)-2'-O-[(tert-butyl)dimethylsilyl]-N 2 - Synthesis of isobutyryl-2'-C-methylguanosine:

[0043]

[0044] In a 250ml two-necked bottle, add 50mlTHF, 50ml pyridine, 0.9g (5mmol) silver nitrate solid and 3.3g (5mmol) 5'-O-(4,4-dimethoxytrityl)-N 2 -Isobutyryl-2'-C-methylguanosine, slowly dropwise add 0.8g (5mmol) of tert-butyldimethylsilyl chloride in pyridine (concentration 1mol / L) under nitrogen protection and ice bath conditions. After continuing stirring for 10 minutes under ice-bath conditions, the mixture was returned to room temperature and stirred for 5 hours. After the TLC plate detected that the reaction was complete, a saturated sodium bicarbonate solution was added in an ice bath until no bubbles emerged, and stirring was continued for 5 minutes. Extracted 3 times with dichloromethane (100mlX3), combined the organic phases, dried over anhydrous sodium sulfate, spin-dried the solvent, loaded the sample by dry method, and separated b...

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Abstract

The invention discloses a 2'-C-methyl guanosine phosphoramidite monomer and a synthesis method thereof. The compound has a structural formula as shown in the specification. The synthesis method of the2'-C-methyl guanosine phosphoramidite monomer comprises reaction steps as shown in the specification. The synthesis method is gentle in reaction condition, simple in aftertreatment and simple and convenient to operate, and conditions are provided for large-scale synthesis of the compounds. The compound disclosed by the invention is a raw material for solid-phase synthesis of RNA (Ribose Nucleic Acid) nucleotide, one or more 2'-C-methyl guanosine phosphoramidite monomers are introduced into an oligonucleotide segment conveniently, and a novel oligonucleotide sequence and a probe thereof can besynthesized. The compound has wide application prospects in research and development of siRNA (small interfering RNA) medicines, study on gene functions, screening of all gene pools, and the like.

Description

technical field [0001] The invention belongs to the technical field of chemical synthesis of nucleotides, in particular to a 2'-C-methylguanosine phosphoramidite monomer and a synthesis method thereof. Background technique [0002] RNA interference technology (RNA interference, RNAi) won the Nobel Prize in Physiology and Medicine in 2006. RNA interference is a gene silencing phenomenon triggered by double-stranded RNA in molecular biology, and its mechanism is to inhibit gene expression by hindering the translation or transcription of specific genes. Small interfering RNA (siRNA) is not only an excellent tool for molecular biology research, but more importantly, it is a potential biomolecule with pharmaceutical application prospects. In recent years, the drug application of siRNA has become a research hotspot in scientific research institutions and enterprises. Among them, the method of chemical modification is a very important method. Although short double-stranded RNA mo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07H19/207C07H1/02C12Q1/6886
CPCC07H19/207C07H1/02C12Q1/6886C12Q2600/178
Inventor 高元和高元盛
Owner 南京百芙利科技有限责任公司
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