# US 2021/0340537 A1 · AN RNA G-QUADRUPLEX STRUCTURE IN PRE-miRNA-1229 AS A THERAPEUTIC TARGET FOR ALZHEIMER'S DISEASE AND VARIOUS CANCERS

> Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt

- Number: US 2021/0340537 A1
- Document ID: US20210340537A1
- Status: Published application
- Kind: A1
- Application number: 17/301,835
- Filed: 2021-04-15
- Published: 2021-11-04
- Earliest claimed: 2020-04-15
- Assignees: Duquesne University of the Holy Spirit
- Applicants: Duquesne University of the Holy Spirit
- Inventors: Mihaela Rita Mihailescu; Joshua A. Imperatore
- Web page: https://patentgrep.com/patent/US20210340537A1

## CPC

- C12N 15/113: Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; {Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing (when used in plants C12N15/8218)}
- C12N 2310/113: targeting other non-coding nucleic acids, e.g. antagomirs
- C12N 2310/3181: Peptide nucleic acid, PNA
- C12N 2310/322: 2'-R Modification
- C12N 2320/34: Allele or polymorphism specific uses
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)

## Abstract

Provided is an agent that binds to a wild type or variant pre-miRNA-1229 comprising a G-quadruplex (GQ) structure, wherein binding of the agent to the wild type or variant pre-miRNA-1229 stabilizes the GQ structure of the wild type or variant pre-miRNA-1229. In some embodiments, the variant is rs2291418. Provided is a method of treating a disease in a subject, comprising: administering a therapeutically effective amount of the agent to the subject. In some embodiments, the disease is Alzheimer's disease, cancer or coronary artery calcification.

## Claims

### Claim 1

An agent that binds to a wild type or variant pre-miRNA-1229 comprising a G-quadruplex (GQ) structure,
wherein binding of the agent to the wild type or variant pre-miRNA-1229 stabilizes the GQ structure of the wild type or variant pre-miRNA-1229.

### Claim 2 (depends on 1)

The agent of claim 1, wherein the variant is rs2291418.

### Claim 3 (depends on 1)

The agent of claim 1, wherein the agent comprises an engineered protein nucleic acid (PNA).

### Claim 4 (depends on 3)

The agent of claim 3, wherein the PNA is a gamma PNA comprising a sequence selected from the group consisting of:
(SEQ ID NO: 5)
AGCCCACGCUCUCCCCCAAA
(SEQ ID NO: 6)
UGAACCCCAGCCCACGCUCU;
and
(SEQ ID NO: 7)
GGGUGUCCCUGAACCCCAGC

### Claim 5 (depends on 3)

The agent of claim 3, wherein the PNA is a gamma PNA comprising a C-terminal lysine residue and comprising a sequence selected from the group consisting of:
(SEQ ID NO: 5)
H2N-Lys AGCCCACGCUCUCCCCCAAA-H
(SEQ ID NO: 6)
H2N-Lys UGAACCCCAGCCCACGCUCU-H;
and
(SEQ ID NO: 7)
H2N-Lys GGGUGUCCCUGAACCCCAGC-H.

### Claim 6 (depends on 1)

The agent of claim 1, wherein the agent comprises 2′-deoxy 2′-fluoroarabino oligonucleotides.

### Claim 7 (depends on 6)

The agent of claim 6, wherein the agent comprises an oligonucleotide sequence selected from the group consisting of:
(SEQ ID NO: 8)
AGCCCACGCUCUCCCCCAAA
(SEQ ID NO: 9)
UGAACCCCAGCCCACGCUCU;
and
(SEQ ID NO: 10)
GGGUGUCCCUGAACCCCAGC.

### Claim 8 (depends on 1)

The agent of claim 1, wherein the agent comprises a small molecule.

### Claim 9 (depends on 1)

A method of treating a disease in a subject, comprising:
administering a therapeutically effective amount of the agent of claim 1 to the subject.

### Claim 10 (depends on 7)

The method of claim 7, wherein the disease is Alzheimer's disease, cancer or coronary artery calcification.

### Claim 11 (depends on 1)

A composition comprising the agent of claim 1.

### Claim 12 (depends on 11)

A method of treating a disease in a subject, comprising:
administering a therapeutically effective amount of the composition of claim 11 to the subject.

### Claim 13 (depends on 12)

The method of claim 12, wherein the disease is Alzheimer's disease, cancer or coronary artery calcification.

### Claim 14

A method of treating Alzheimer's disease, cancer or other disease and mitigating their progression, comprising:
stabilizing or destabilizing a novel therapeutic target, comprising:
a pre-miRNA-1229 sequence, comprising:
a G-quadruplex structure; and
a canonical hairpin structure,
wherein the G-quadruplex and the canonical hairpin structures co-exist in equilibrium in the guanine-rich region of the pre-miRNA-1229 sequence;
controlling the production of mature miRNA-1229-3p; and
correspondingly regulating sortilin-related receptor protein.

### Claim 15 (depends on 14)

The method of claim 14, wherein the regulating sortilin-related receptor protein comprises upregulation of expression of the sortilin-related receptor for Alzheimer's disease.

### Claim 16 (depends on 14)

The method of claim 14, wherein the stabilizing or destabilizing of the G-quadruplex structure includes a single nucleotide polymorphism rs2291418 effective to change the equilibrium between the G-quadruplex structure and the canonical hairpin structure.

### Claim 17 (depends on 16)

The method of claim 16, wherein the change in equilibrium favors the canonical hairpin structure.

### Claim 18 (depends on 14)

The method of claim 14, wherein for Alzheimer's disease, the method comprises stabilizing the G-quadruplex structure, lowering the production of mature miRNA-1229-3p to a level corresponding to a normal level, and increasing the SORL1 protein to a level corresponding to a normal level.

### Claim 19

A method of controlling miRNA dysregulation to reduce effects of Alzheimer's disease, cancer or other disease, comprising:
targeting a G-quadruplex structure in equilibrium with a canonical hairpin structure in a guanine-rich region of a pre-miRNA-1229 sequence;
regulating rs2291418 single nucleotide polymorphism located in the pre-miRNA 1229 sequence;
changing the equilibrium between the G-quadruplex structure and the canonical hairpin structure to favor the G-quadruplex structure or the canonical hairpin structure;
controlling the production of mature miRNA-1229-3p; and
correspondingly upregulating or downregulating a sortilin-related receptor protein.

### Claim 20 (depends on 19)

The method of claim 19, wherein for Alzheimer's disease, the method comprises stabilizing the G-quadruplex structure to favor the G-quadruplex, lowering the production of mature miRNA-1229-3p to a level corresponding to a normal level and upregulating the sortilin-related receptor protein to a level corresponding to a normal level.