siRNA for inhibiting expression of CTGF gene, pharmaceutical composition containing siRNA and use of pharmaceutical composition

A gene expression and base technology, applied in the field of nucleic acid and pharmaceutical composition, can solve the problems of poor stability of siRNA activity, slow progress, slow progress of drugs, etc.

Pending Publication Date: 2020-07-07
SUZHOU RIBO LIFE SCIENCE CO LTD
View PDF4 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, so far, the clinical application of siRNA drugs for the treatment of diseases related to CTGF gene expression has been slow. Amo

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
  • siRNA for inhibiting expression of CTGF gene, pharmaceutical composition containing siRNA and use of pharmaceutical composition
  • siRNA for inhibiting expression of CTGF gene, pharmaceutical composition containing siRNA and use of pharmaceutical composition
  • siRNA for inhibiting expression of CTGF gene, pharmaceutical composition containing siRNA and use of pharmaceutical composition

Examples

Experimental program
Comparison scheme
Effect test

Example Embodiment

[0235] Example

[0236] Unless otherwise specified, the reagents and culture media used in the following examples are all commercially available products. The nucleic acid electrophoresis, real-time PCR and other operations used are all referred to the methods described in Molecular Cloning (Cold Spring Harbor Laboratory Press (1989)) get on.

[0237] Hela cells were provided by the Nucleic Acid Technology Laboratory of the Institute of Molecular Medicine, Peking University, with 20% fetal bovine serum (FBS, Hyclone) and 0.2% by volume penicillin double antibody (Penicillin-Streptomycin, Gibco, Invitrogen) Culture cells in complete DMEM medium (Hyclone) at 37℃ in 5% CO 2 / Cultivate in an incubator with 95% air.

[0238] If there is no other instructions, the reagent ratios provided below are calculated by volume ratio (v / v).

Example Embodiment

[0239] Preparation Example 1 Synthesis of siRNA sequence

[0240] In siRNA synthesis, unless otherwise specified, the nucleoside monomer refers to the modified nucleoside phosphoramidite used in the solid phase synthesis of phosphoramidites according to the type and sequence of nucleotides in the siRNA to be prepared Monomers (modified RNA phosphoramidites). Phosphoramidite solid-phase synthesis is a method used in RNA synthesis well known to those skilled in the art. Unless otherwise specified, all nucleoside monomers used are commercially available.

[0241] (1-a) Solid phase phosphoramidite method

[0242] The siRNA sequences listed in Table 2 were obtained by solid phase phosphoramidite method.

[0243] For the justice chain, a universal solid phase carrier (UnyLinker TM loaded HL SolidSupports, Kinovate Life Sciences, Inc.) starts the cycle, and connects the nucleoside monomers one by one from the 3'-5' direction according to the sequence of nucleotide arrangement. Each conn...

Example Embodiment

[0283] Experimental Example 1 Detection of the inhibitory efficiency of siRNA on CTGF mRNA expression in Hela cells.

[0284] Use Lipofectamine TM In 2000, the siRNAs obtained in Preparation Example 1 were respectively transfected into Hela cells, and the final concentration of siRNA was 50 nM. Each siRNA was transfected into 3 replicate wells. Cells without any siRNA treatment served as blank control.

[0285] The expression of CTGF mRNA in Hela cells transfected with each siRNA was detected by Quantitative Real-Time PCR. The specific steps are: After culturing the transfected cells for 24 hours, use Trizol (Thermo Fisher) to extract the total RNA from the cells according to the standard operating procedures for total RNA extraction; take 1 μg of total RNA, and use the reverse transcription kit (Promega) , Item No. A3500) Reverse transcription to obtain cDNA according to the operating method of its manual. Use 2×Ultra SYBR Mixture (with ROX) (Beijing Kangwei Century Biotechnol...

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

The invention relates to siRNA for inhibiting expression of a CTGF gene. The siRNA contains a positive-sense strand and an antisense strand, wherein the positive-sense strand contains a nucleotide sequence I, the antisense strand contains a nucleotide sequence II, the nucleotide sequence I and the nucleotide sequence II are both 19 nucleotides in length, and the nucleotide sequence II at least partially and mutually complements a sequence which is 19 nucleotides in length in a region from site No. 700 to site No. 1050 of CTGF mRNA. From a terminal 5' to a terminal 3', nucleotides at sites No.7, No. 8 and No. 9 of the nucleotide sequence I are fluoro-modified nucleotides, and nucleotides at sites No. 2, No. 6, No. 14 and No. 16 of the nucleotide sequence II are fluoro-modified nucleotides.The invention further provides a pharmaceutical composition containing the siRNA. The siRNA and the pharmaceutical composition containing the siRNA can be used for treating or improving diseases relevant with the expression of the CTGF gene.

Description

technical field [0001] The disclosure relates to a nucleic acid capable of inhibiting CTGF gene expression and a pharmaceutical composition containing the nucleic acid, belonging to the field of nucleic acid pharmacy. The present disclosure also relates to the use of these nucleic acids and pharmaceutical compositions. Background technique [0002] Liver fibrosis refers to the excessive deposition of diffuse extracellular matrix (ECM) in the liver. It is a compensatory response in the tissue repair process secondary to various forms of chronic liver injury, and it is also the cause of chronic liver disease that develops into cirrhosis, liver cancer, etc. Severe fatal diseases must go through the pathological process, so anti-hepatic fibrosis has become the top priority in the treatment of chronic liver diseases. [0003] At present, the means of treating liver fibrosis are very limited, mainly including two aspects: one is to remove the pathogenic factors for the primary di...

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
IPC IPC(8): C12N15/113A61K31/713A61P1/16A61P13/12A61P11/00A61P1/00A61P9/00A61P9/10A61P17/00A61P27/02
CPCC12N15/113A61K31/713A61P1/16A61P13/12A61P11/00A61P1/00A61P9/00A61P9/10A61P17/00A61P27/02C12N2310/14
Inventor 张鸿雁高山康代武郑书全
Owner SUZHOU RIBO LIFE SCIENCE CO LTD
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

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

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap