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Nucleic acid molecule capable of inhibiting expression of periostin gene, method for inhibiting expression of periostin gene, and use of said nucleic acid molecule

A nucleic acid molecule, periostin technology, applied in gene therapy, genetic engineering, biochemical equipment and methods, etc., can solve problems such as blindness

Inactive Publication Date: 2014-12-17
KYUSHU UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

And, if this state continues, repeats and becomes serious, blindness may be caused (Patent Document 1)

Method used

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  • Nucleic acid molecule capable of inhibiting expression of periostin gene, method for inhibiting expression of periostin gene, and use of said nucleic acid molecule
  • Nucleic acid molecule capable of inhibiting expression of periostin gene, method for inhibiting expression of periostin gene, and use of said nucleic acid molecule
  • Nucleic acid molecule capable of inhibiting expression of periostin gene, method for inhibiting expression of periostin gene, and use of said nucleic acid molecule

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0462] siRNA was synthesized, and expression suppression of the human periostin gene in vitro was confirmed.

[0463] (1) siRNA

[0464]As an example of siRNA, a double-stranded RNA having the following sequence was synthesized. In each double-stranded RNA, the upper sequence is the sense strand, and the lower sequence is the antisense strand. The 3' overhang of the sense strand and the 3' overhang of the antisense strand are both n=2, which is TT.

[0465] [Table 7]

[0466]

[0467] In addition, as a negative control, the following double-stranded RNA in which the base sequence of the antisense strand was randomly scrambled (scrambled) was used.

[0468] [Table 8]

[0469] NI-0000

[0470] (SEQ ID NO: 77) 5'-UACUAUUCGACACGCGAAGUU-3'

[0471] (Serial No. 78) 3'-UUAUGAUAAGCUGUGCGCUUC-5'

[0472] (2) Determination of the expression level of periostin gene

[0473] As cells, human retinal pigment epithelial cell line ARPE-19 (American Type Culture Collection: ATCC...

Embodiment 2

[0486] Using a choroidal neovascularization (CNV) model mouse, it was confirmed that the formation of neovascularization and fibrous proliferative tissue was inhibited by the nucleic acid molecule of the present invention.

[0487] (1) Nucleic acid molecule

[0488] As the siRNA of the example, NI-0079 of the above-mentioned Example 1 was used. In addition, NK-0144 and PK-0076 shown below were used as single-stranded nucleic acid molecules in Examples. In NK-0144 and PK-0076, the as1 nucleotide of SEQ ID NO: 1 common to the antisense strand of the siRNA is underlined.

[0489] [Table 9]

[0490]

[0491] In the above-mentioned PK-0076, the structure of the linker (Lx) and the linker (Ly) is the following structure shown in the above-mentioned (I-8a).

[0492]

[0493] As a negative control, the same NI-0000 as in Example 1 was used.

[0494] (2) Production of model mice

[0495] Mix Ketalar (registered trademark) injection (50 mg / mL) and Selactar (registered tradema...

Embodiment 3

[0509] siRNA was synthesized, and expression suppression of the human periostin gene in vitro was confirmed.

[0510] As an example of siRNA, a double-stranded RNA having the following sequence was synthesized. In each double-stranded RNA, the upper sequence is the sense strand, and the lower sequence is the antisense strand. The 3' overhang of the sense strand and the 3' overhang of the antisense strand were both n=2. In addition, relative gene expression levels were measured in the same manner as in Example 1 above except that these siRNAs were used.

[0511] [Table 10]

[0512]

[0513] show these results in Figure 7 . Figure 7 It is a graph showing the relative value of periostin gene expression level, and the vertical axis is the relative gene expression level. Such as Figure 7 As shown, the double-stranded RNA of the Example showed lower values ​​than the control (- and mock) and the negative control, and thus it was possible to confirm that both had expressi...

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PUM

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Abstract

Provided is a novel molecule which can inhibit the expression of periostin gene. A nucleic acid molecule capable of inhibiting the expression of periostin gene is characterized by containing nucleotides that constitute at least one nucleotide sequence selected from the nucleotide sequences represented by SEQ ID NOs: 1 to 19 as a sequence capable of inhibiting the expression of periostin gene. The nucleic acid molecule capable of inhibiting the expression according to the present invention can inhibit the expression of periostin gene. Therefore, the nucleic acid molecule can be used for the treatment of eye diseases associated with the expression of periostin gene, specifically proliferative diabetic retinopathy, macular degeneration and the like.

Description

technical field [0001] The present invention relates to a nucleic acid molecule for inhibiting expression of periostin gene, a method for inhibiting expression of periostin gene, and uses thereof. Background technique [0002] In recent years, the number of patients with diabetic retinopathy (Diabetic Retinopathy: DR), which is one of the complications, is increasing along with the increase in diabetic patients. Diabetic retinopathy is classified into simple diabetic retinopathy, proliferative diabetic retinopathy, and proliferative diabetic retinopathy (Proliferative Diabetic Retinopathy: PDR) according to disease stages. Among them, proliferative diabetic retinopathy not only has decreased vision, but also may lead to blindness. [0003] In addition, as an eye disease, macular degeneration such as age-related macular degeneration (Age-related Macular Degeneration: AMD) is also a problem. Macular degeneration is a disease that causes macular degeneration and vision loss d...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/09A61K31/7088A61K48/00A61P27/02A61P43/00C12N15/113
CPCC12N2310/11C12N2310/14C12N2310/532A61K31/7105A61K31/713C12N15/113A61P27/02A61P43/00
Inventor 吉田茂生滨崎智洋石川桂二郎水谷隆之石桥达朗大木忠明中间崇仁
Owner KYUSHU UNIV
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