# US 12,758,573 B2 · RNA action inhibitor and use thereof

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- Number: US 12,758,573 B2
- Document ID: US12758573B2
- Status: Granted
- Kind: B2
- Application number: 17/637,707
- Filed: 2020-08-20
- Published: 2022-12-22
- Granted: 2026-10-13
- Earliest claimed: 2019-08-23
- Assignees: National University Corporation Tokai; National Higher Education and Research System; Nicca Chemical Co., Ltd. Hokkaido System Science Co., Ltd.
- Applicants: National University Corporation Tokai National Higher Education and Research System; Nicca Chemical Co., Ltd.; Hokkaido System Science Co., Ltd.
- Inventors: Hiroyuki Asanuma; Yukiko Kato; Keiji Murayama; Toshiki Tsuboi; Tomohito Doke; Takuji Ishimoto; Shoichi Maruyama; Toshio Kokuryo; Ayako Koto; Isao Ishimaru; Ryoichi Asai; Ayaka Nakasha; Tomoe Sugita; Keisuke Nishi; Motoki Yuguchi; Masanori Matsumoto
- Web page: https://patentgrep.com/patent/US12758573B2

## CPC

- A61K 31/7115: Nucleic acids or oligonucleotides having modified bases, i.e. other than adenine, guanine, cytosine, uracil or thymine
- C12N 15/1138: {against receptors or cell surface proteins}
- A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D)
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)

## Abstract

An agent for inhibiting the action of a target RNA based on RNA interference and antisense oligonucleotide (ASO). The RNA interference agent includes a passenger strand and a guide strand for a target RNA. Both ends of double-stranded RNA formed pairing of the passenger and guide strands are blunt ends, and one or two or more of the units represented by formula (1) and formula (2) below are provided at the following positions (a) and (b): (a) 5′ end side and 3′ end side of the passenger strand, (b) 3′ end side of the guide strand where X represents an oxygen atom or sulfur atom, where X represents an oxygen atom or sulfur atom.

## Claims

### Claim 1

An RNA interference agent comprising a passenger strand and a guide strand for a target RNA,
wherein both ends of double-stranded RNA formed by pairing of the passenger strand and the guide strand constitute blunt ends, and
wherein the passenger strand comprises two units represented by formula (2) at each of 5′ end side and 3′ end side and the guide strand comprises two units represented by formula (2) at the 3′ end side,
where X represents an oxygen atom or sulfur atom.

### Claim 2 (depends on 1)

The RNA interference agent according to claim 1, wherein the target RNA is mRNA of NeK2 gene.

### Claim 3 (depends on 2)

The RNA interference agent according to claim 2, wherein the passenger strand has the base sequence represented by SEQ ID NO:9, and the guide strand has the base sequence represented by SEQ ID NO:10.

### Claim 4

A method for suppressing the expression of a target gene, comprising:
administering an RNA duplex to a subject or an expression system in vivo, in vitro or ex vivo, the RNA duplex comprising a passenger strand and a guide strand for a target RNA that is at least a portion of mRNA of the target gene,
wherein both ends of the double-stranded RNA formed by pairing the passenger strand and the guide strand form blunt ends,
wherein the passenger strand comprises two units represented by formula (2) at each of the 5′ and 3′ ends, and the guide strand comprises two units represented by formula (2) at the 3′ end,
where X represents an oxygen atom or a sulfur atom.

### Claim 5

A target RNA detection agent comprising an antisense strand for the target RNA, the antisense strand comprising:
a hybridization region comprising units represented by formula (1) capable of specifically hybridizing to the target RNA;
a stem region comprising units represented by formula (1), directly linked to 3′ end and 5′ end of the hybridization region, respectively, and hybridizing to form a loop around the hybridization region;
a signal element indicating that the hybridization region has hybridized to the target RNA; and
one or two diaminopurine-containing units having 2,6-diaminopurine as a base instead of one or two adenine-units having an adenine base in the hybridization region, the adenine-containing units being spaced one to three bases from the 3′ end and 5′ end of the hybridization region,
where X represents an oxygen atom or a sulfur atom.

### Claim 6 (depends on 5)

The target RNA detection agent of claim 5, wherein the target RNA is miR21.

### Claim 7 (depends on 6)

The target RNA detection agent of claim 6, wherein the hybridizing region and the stem region have a continuous base sequence represented by SEQ ID NO: 21.

### Claim 8 (depends on 5)

The target RNA detection agent of claim 5, wherein the target RNA is mRNA of EGFR gene and is for detecting a single-base substitution in the gene.

### Claim 9 (depends on 8)

The target RNA detection agent of claim 8, wherein the hybridizing region and the stem region have a continuous base sequence selected from the base sequences represented by SEQ ID NOs: 14 to 20.