Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Inhibition Of Viral Gene Expression

A virus and hepatitis virus technology, applied in gene therapy, antiviral agents, genetic engineering, etc., can solve the problems of enlarged synthesis reaction, low yield and difficulty of multi-step chemical synthesis

Inactive Publication Date: 2014-08-27
UNIVERSITY OF THE WITWATERSRAND +1
View PDF2 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

While encouraging, one problem with utilizing these siRNA reagents is that the yields of the multi-step chemical synthesis are rather low
In addition, it is difficult to scale up the synthetic reaction

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Inhibition Of Viral Gene Expression
  • Inhibition Of Viral Gene Expression
  • Inhibition Of Viral Gene Expression

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0063] Example 1 Synthesis of four kinds of 2'-O-guanidinopropyl-nucleoside-phosphoramidites

[0064] Each of the four 2'-O-guanidinopropyl-nucleoside-phosphoramidites was synthesized using essentially similar methods. figure 1 The synthesis of adenosine (A), cytidine (C) and uridine (U) derivatives is described. Separately in the diagram ( figure 2 )show. Each synthesis starts with the use of 1,1,3,3-tetraisopropyldisiloxane-1,3-diyl (TIPS) (for A, C and U) or di-tert-butylsilanediyl ( DTBS) (for guanosine) protects both the 5'- and 3'-hydroxyl groups. DTBS was chosen to protect G because this group has been reported to increase the selectivity of subsequent 2,4,6-triisopropylbenzenesulfonyl (TPS) protection at the O6-position of guanosine [21]. The exocyclic amino functions of A and C were protected with a dimethylaminomethylene group using standard conditions, and a benzoyl group was attached at the N3-position of U using a two-phase system reported by Sekine [22]. Th...

Embodiment 2

[0069] Example 2 Synthesis of 2'-O-guanidinopropyl adenosine phosphoramidite

[0070] 3′,5′-O-(tetraisopropyldisiloxane-1,3-diyl)-N6-dimethylaminomethyleneadenosine (1a) was synthesized as previously described [16].

[0071] N6-Dimethylaminomethylene-2′-O-cyanoethyl-3′,5′-O-(tetraisopropyldisiloxane-1,3-diyl)-adenosine (1e)

[0072] Freshly distilled acrylonitrile (6.7 mL, 102 mmol) and cesium carbonate (1.6 g, 4.9 mmol) were added to a solution of compound la (3.0 g, 5.31 mmol) in tert-butanol (25 mL). The mixture was stirred vigorously at room temperature for 3 h. The reaction mixture was filtered and the residue was washed with dichloromethane. The filtrate was evaporated and the residue was purified by column chromatography with ethyl acetate / methanol (99:1 - 95:5, v / v) yielding 3.28 g (87%) of product. 1H NMR (400MHz, DMSO-d6) δ [ppm] 8.90 (s, 1H, amidine-H), 8.34 (s, 1H, H2 or H8), 8.32 (s, 1H, H2 or H8), 6.02-6.01 ( m,1H,H1′),5.05-5.01(m,1H,H3′),4.64-4.62(m,1H,H2′),...

Embodiment 3

[0087] Example 3 Synthesis of 2'-O-guanidinopropylcytidine phosphoramidite

[0088] N4-Dimethylaminomethylene-3′,5′-O-(tetraisopropyldisiloxane-1,3-diyl)-cytidine (2a) according to the previously described reaction procedure [29 ]synthesis.

[0089] N4-Dimethylaminomethylene-2′-O-cyanoethyl-3′,5′-O-(tetraisopropyldisiloxane-1,3-diyl)-cytidine (2e)

[0090] Compound 2a (4 g, 7.39 mmol) was dissolved in acrylonitrile (8 mL, 122 mmol) and tert-butanol (35 mL). Cesium carbonate (1.8 g, 5.52 mmol) was added and the reaction mixture was stirred at room temperature for 2.5 h. The mixture was filtered through celite, and the residue was purified by column chromatography after evaporation of the solvent. Ethyl acetate was used initially as solvent, which was changed to ethyl acetate / methanol (9:1, v / v) after the non-polar mixture was passed through the column. The product was obtained in a yield of 3.78 g (86%). 1H NMR (400MHz, DMSO-d6) δ [ppm] 8.62 (s, 1H, N4 = CH-NMe2), 7.88 (d,...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

This invention relates to modified short interfering RNA (siRNA) nucleic acid molecules, particularly siRNA's which have been modified by the addition of a 2 -0- guanidinopropyl (GP) modified nucleoside. In particular the invention relates to modified siRNAs which are capable of silencing target sequences, methods of treating and preventing infection by using the siRNAs, medicaments containing the siRNAs and use of the siRNAs.

Description

[0001] introduction [0002] The present invention relates to modified small interfering RNA molecules (siRNA) that regulate gene expression through the RNA interference pathway. Nucleic acid molecules encoding siRNAs of the invention include one or more modifications that cause their physical properties to differ from wild-type unmodified siRNAs. In a preferred embodiment of the present invention, the nucleic acid sequence of the siRNA includes at least one nucleoside with a 2-O-guanidinopropyl (GP) structure. In a further embodiment of the present invention, the modification of the siRNA improves the stability of the modified siRNA, promotes gene silencing caused by the modified siRNA and reduces immune stimulation. Background technique [0003] Synthetic activators of RNA interference (RNAi) show considerable potential for therapeutic applications in pathological gene silencing. Some typical exogenous RNA interference activators include approximately 21 bp double-stranded...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/113A61K31/712A61P31/20
CPCA61K31/713C12N15/1131C12N2310/3527C12N2310/321C12N2320/30C12N15/113A61P31/20
Inventor 帕特里克·阿巴思诺特贾斯廷·希恩阿布杜拉·伊利穆萨·马里亚尼约兰塔·布瑞恩斯卡詹尼弗·多诺费奥马克西米利安C.R.·布夫约阿希姆W.·恩格斯斯蒂芬·贝恩哈特
Owner UNIVERSITY OF THE WITWATERSRAND
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com