Acetyl-coa carboxylase2 antisense oligonucleotides

A peptide nucleic acid and compound technology, applied in the field of peptide nucleic acid derivatives, can solve the problems of poor therapeutic activity and poor cell permeability, and achieve the effect of good cell permeability

Pending Publication Date: 2021-03-26
OLIPASS CORP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Like other oligonucleotide classes with DNA or RNA backbones, siRNAs are poorly cell permeable and thus tend to exhibit poor in vitro or in vivo therapeutics unless properly formulated or chemically modified to have good membrane permeability active

Method used

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  • Acetyl-coa carboxylase2 antisense oligonucleotides
  • Acetyl-coa carboxylase2 antisense oligonucleotides
  • Acetyl-coa carboxylase2 antisense oligonucleotides

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0186] Example 1: "ASO 1"-induced exon skipping in C2C12

[0187] The ability of "ASO 1" to induce ACC2 "exon 12" skipping in C2C12 cells was assessed as described below.

[0188] [Cell culture and ASO treatment] In DMEM medium supplemented with 10% FBS (fetal bovine serum) (Cat. No. 10099-41, GIBCO) and 1% streptomycin / penicillin (Cat. No. 15140-122, GIBCO) (Dulbecco's Modified Eagle's Medium: DMEM) (Cat. No. 12-604F, Lonza) in 60 mm Petri dishes at 5% CO 2 Grow C2C12 cells (2x10 5 ) (Cat. No. CRL-1772, ATCC). Cells were either left untreated (negative control) or treated with an aliquot of "ASO 1" aqueous stock stock at 100 zM to 1 fM for 5 hours.

[0189] [RNA extraction and nested PCR] Total RNA was extracted from "ASO 1"-treated cells using the RNeasy Mini Kit (Qiagen, Cat. No. 714106) according to the manufacturer's TM III One-step RT-PCR system (Catalog No. 12574-018, Invitrogen) prepared cDNA from 200 ng RNA. Add 200ngRNA, 25μl 2X Reaction Mix buffer, 2μl SuperSc...

Embodiment 2

[0192] Example 2: Inhibition of ACC2 mRNA Formation by "ASO 1" in C2C12

[0193] The ability of "ASO 1" to downregulate ACC2 mRNA formation in C2C12 was assessed by real-time qPCR as described below.

[0194] [Cell Culture & ASO Treatment] C2C12 cells (Cat. No. CRL-1772, ATCC) were maintained in a culture medium supplemented with 10% fetal bovine serum (Cat. No. 10099-41, GIBCO) and 1% streptomycin / penicillin (Cat. No. 15140-122 , GIBCO) in Dulbecco's Modified Eagle Medium (DMEM, Cat. No. 12-604F, Lonza), at 37°C and 5% CO 2 grow under conditions. C2C12 cells (2×10 5 ) were incubated with 0 (negative control) and 100 zM to 1 fM of "ASO 1" for 24 hours.

[0195] [RNA extraction & cDNA synthesis] Total RNA was extracted from "ASO 1"-treated cells using the RNeasy Mini Kit (Qiagen, Cat. No. 714106) according to the manufacturer's instructions and analyzed using PrimeScript TM 1 st cDNA was prepared from 400 ng of RNA with a strand cDNA synthesis kit (Takara, Cat# 6110A). ...

Embodiment 3

[0198] Example 3: Inhibition of ACC2 mRNA Formation by "ASO 6" in C2C12

[0199] The ability of "ASO 6" to downregulate ACC2 mRNA formation in C2C12 was assessed by real-time qPCR as described below.

[0200] [Cell Culture & ASO Treatment] C2C12 cells (Cat. No. CRL-1772, ATCC) were maintained in a culture medium supplemented with 10% fetal bovine serum (Cat. No. 10099-41, GIBCO) and 1% streptomycin / penicillin (Cat. No. 15140-122 , GIBCO) in Dulbecco's Modified Eagle Medium (DMEM, Cat. No. 12-604F, Lonza), at 37°C and 5% CO 2 grow under conditions. C2C12 cells (2×10 5 ) were incubated with 0 (negative control) and 100 zM to 1 fM of "ASO 6" for 24 hours.

[0201] [RNA Extraction & cDNA Synthesis] Total RNA was extracted from "ASO 6" treated cells using RNeasy Mini Kit (Qiagen, Cat. No. 714106) according to the manufacturer's instructions and analyzed using PrimeScript TM 1 st cDNA was prepared from 400 ng of RNA with a strand cDNA synthesis kit (Takara, Cat# 6110A). To a...

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Abstract

The present invention provides the peptide nucleic acid derivative which targets 5' splice site of the human ACC2 pre-mRNA "exon 12". The peptide nucleic acid derivatives in the present invention strongly induce splice variants of the human ACC2 mRNA in cell and are very useful to treat conditions or disorders of skin aging associated with the human ACC2 protein.

Description

technical field [0001] The present invention relates to peptide nucleic acid derivatives that complementarily target human acetyl-CoA carboxylase 2 precursor mRNA (pre-mRNA) to improve skin aging mediated by acetyl-CoA carboxylase 2. Background technique [0002] Skin aging has received considerable attention since the signs of aging are most visible in the skin. Skin aging begins in the mid to late 20s, when collagen and elastin in the skin decrease, resulting in dry skin with low elasticity and even wrinkles. Obesity is an inflammatory response caused by decreased blood circulation caused by excess fat deposits in the body. Internal fat on blood vessels inhibits blood circulation and the secretion of various hormones, thereby promoting the aging of the entire body including the skin. In this sense, obesity-related health conditions and diseases must be monitored for healthy and beautiful skin. [0003] The biosynthesis and degradation of fatty acids are well regulated a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/00A61P17/00A61Q19/08A61K38/00C12N15/113A61K8/64
CPCA61K8/64A61K38/00A61P17/00A61Q19/08C12Y604/01002C12N2310/3181C12N2310/11C12N15/1137A61K38/53C12N9/93C07K14/003
Inventor 韩璿瑛成基浩洪铭涍姜多映许情奭张降愿
Owner OLIPASS CORP
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