# US 2008/0306014 A1 · Agents, compositions and methods for treating pathologies in which regulating an ache-associated biological pathway is beneficial

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- Number: US 2008/0306014 A1
- Document ID: US20080306014A1
- Status: Published application
- Kind: A1
- Application number: 11/714,861
- Filed: 2007-03-07
- Published: 2008-12-11
- Earliest claimed: 2004-09-07
- Assignees: Yissum Research Development Company of the Hebrew University of Jerusalem
- Inventors: Hermona Soreq; Iftach Shaked; Ran Avni; Ari Meerson
- Web page: https://patentgrep.com/patent/US20080306014A1

## 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)}
- A61P 43/00: Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
- C12N 15/111: {General methods applicable to biologically active non-coding nucleic acids}
- C12N 2310/11: Antisense
- C12N 2310/14: interfering nucleic acids [NA]
- C12N 2310/321: 2'-O-R Modification
- C12N 2320/50: Methods for regulating/modulating their activity
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS

## Abstract

The present invention provides agents which are capable of regulating the function of a micro-RNA component which can be used to regulate an AChE-associated biological pathway. In addition, the present invention provides methods and pharmaceutical compositions for the treatment of various pathologies related to AChE-associated biological pathways such as apoptosis, aberrant cholinergic signaling, abnormal hematopoietic proliferation and/or differentiation, cellular stress, exposure to inflammatory response-inducing agents, and/or exposure to organophosphates or other AChE inhibitors.

## Claims

### Claim 1

A method of regulating an AChE-associated biological pathway having a miRNA component, the method comprising subjecting the AChE-associated biological pathway to an agent capable of regulating a function of the miRNA, thereby regulating the AChE-associated biological pathway.

### Claim 2 (depends on 1)

The method of claim 1, wherein said agent is a polynucleotide.

### Claim 3 (depends on 2)

The method of claim 2, wherein said polynucleotide is selected from the group consisting of a polynucleotide which comprises at least 10 consecutive nucleotides of the nucleic acid sequence set forth in SEQ ID NO:1, a polynucleotide hybridizable in cells under physiological conditions to an RNA molecule which comprises a nucleic acid sequence as set forth in SEQ ID NO:2, a polynucleotide as set forth by SEQ ID NO:1, a polynucleotide which comprises at least 10 consecutive nucleotides of the nucleic acid sequence set forth in SEQ ID NO:2, a polynucleotide hybridizable in cells under physiological conditions to an RNA molecule which comprises a nucleic acid sequence as set forth in SEQ ID NO:21 and/or 22, a polynucleotide as set forth by SEQ ID NO:2, a polynucleotide, which comprises at least 25 consecutive nucleotides of the nucleic acid sequence set forth in SEQ ID NO:13, a polynucleotide as set forth by SEQ ID NO:13, a polynucleotide which comprises at least 20 consecutive nucleotides of SEQ ID NO:13 and/or at least 10 consecutive nucleotides of SEQ ID NO:1, a polynucleotide as set forth by SEQ ID NO:12 or a functional homolog thereof, a polynucleotide as set forth by SEQ ID: 19 or a functional homolog thereof, a polynucleotide as set forth by SEQ ID NO:23 and a polynucleotide as set forth by SEQ ID NO: 24.

### Claim 4 (depends on 1)

The method of claim 1, wherein said miRNA is set forth by the sequence selected from the group consisting of SEQ ID NOs: 54, 93, 94, 98, 99, 100, 21 and 22.

### Claim 5

A method of regulating an expression level ratio of AChE-S and AChE-R and/or AChE-S mRNA and AChE-R mRNA splice variants in AChE expressing cells comprising subjecting the AChE gene expressing cells to an agent capable of regulating a function of a miRNA component associated with regulating the expression level ratio of AChE-S and AChE-R splice variants, thereby regulating the expression level of the AChE-S and AChE-R splice variants in the AChE expressing cells.

### Claim 6 (depends on 5)

The method of claim 5, wherein said agent is a polynucleotide.

### Claim 7 (depends on 6)

The method of claim 6, wherein said polynucleotide is selected from the group consisting of a polynucleotide which comprises at least 10 consecutive nucleotides of the nucleic acid sequence set forth in SEQ ID NO:1, a polynucleotide hybridizable in cells under physiological conditions to an RNA molecule which comprises a nucleic acid sequence as set forth in SEQ ID NO:2, a polynucleotide as set forth by SEQ ID NO:1, a polynucleotide which comprises at least 10 consecutive nucleotides of the nucleic acid sequence set forth in SEQ ID NO:2, a polynucleotide hybridizable in cells under physiological conditions to an RNA molecule which comprises a nucleic acid sequence as set forth in SEQ ID NO:21 and/or 22, a polynucleotide as set forth by SEQ ID NO:2, a polynucleotide which comprises at least 25 consecutive nucleotides of the nucleic acid sequence set forth in SEQ ID NO:13, a polynucleotide as set forth by SEQ ID NO:13, a polynucleotide which comprises at least 20 consecutive nucleotides of SEQ ID NO:13 and/or at least 10 consecutive nucleotides of SEQ ID NO:1, a polynucleotide as set forth by SEQ ID NO:12 or a functional homolog thereof, a polynucleotide as set forth by SEQ ID: 19 or a functional homolog thereof, a polynucleotide as set forth by SEQ ID NO:23 and a polynucleotide as set forth by SEQ ID NO: 24.

### Claim 8 (depends on 4)

The method of claim 4, wherein said miRNA is set forth by the sequence selected from the group consisting of SEQ ID NOs: 54, 93, 94, 98, 99, 100, 21 and 22.

### Claim 9

A method of treating a pathology related to an AChE-associated biological pathway, the method comprising administering to a subject in need thereof an agent capable of regulating a function of a miRNA component of the AChE-associated biological pathway, thereby treating the pathology.

### Claim 10 (depends on 9)

The method of claim 9, wherein the pathology is a disease or condition in which regulating nitric oxide levels is therapeutically beneficial.

### Claim 11 (depends on 9)

The method of claim 9, wherein the pathology is associated with abnormal levels of AChE-S or AChE-R splice variants.

### Claim 12 (depends on 9)

The method of claim 9, wherein said polynucleotide is selected from the group consisting of a polynucleotide which comprises at least 10 consecutive nucleotides of the nucleic acid sequence set forth in SEQ ID NO:1, a polynucleotide hybridizable in cells under physiological conditions to an RNA molecule which comprises a nucleic acid sequence as set forth in SEQ ID NO:2, a polynucleotide as set forth by SEQ ID NO:1, a polynucleotide which comprises at least 10 consecutive nucleotides of the nucleic acid sequence set forth in SEQ ID NO:2, a polynucleotide hybridizable in cells under physiological conditions to an RNA molecule which comprises a nucleic acid sequence as set forth in SEQ ID NO:21 and/or 22, a polynucleotide as set forth by SEQ ID NO:2, a polynucleotide which comprises at least 25 consecutive nucleotides of the nucleic acid sequence set forth in SEQ ID NO:13, a polynucleotide as set forth by SEQ ID NO:13, a polynucleotide which comprises at least 20 consecutive nucleotides of SEQ ID NO:13 and/or at least 10 consecutive nucleotides of SEQ ID NO:1, a polynucleotide as set forth by SEQ ID NO:12 or a functional homolog thereof, a polynucleotide as set forth by SEQ ID: 19 or a functional homolog thereof, a polynucleotide as set forth by SEQ ID NO:23 and a polynucleotide as set forth by SEQ ID NO: 24.

### Claim 13 (depends on 9)

The method of claim 9, wherein said miRNA is set forth by the sequence selected from the group consisting of SEQ ID NOs: 54, 93, 94, 98, 99, 100, 21 and 22.

### Claim 14

A method of altering differentiation and/or proliferation of hematopoietic progenitor and/or stem cells, the method comprising subjecting the progenitor and/or stem cells to an agent capable of regulating a function a miRNA component of an AChE-associated biological pathway in the progenitor and/or stem cells, thereby altering differentiation and/or proliferation of the hematopoietic progenitor and/or stem cells.

### Claim 15

A method of regulating apoptosis in cells and/or a tissue of a subject in need thereof, the method comprising subjecting the cells and/or the tissue of the subject to an agent capable of regulating a function a miRNA component of an AChE-associated biological pathway in the cells and/or tissue, thereby regulating apoptosis in the cells and/or the tissue of the subject.

### Claim 16 (depends on 15)

The method of claim 15, wherein said agent is a polynucleotide.

### Claim 17 (depends on 16)

The method of claim 16, wherein said polynucleotide is selected from the group consisting of a polynucleotide which comprises at least 10 consecutive nucleotides of the nucleic acid sequence set forth in SEQ ID NO:1, a polynucleotide hybridizable in cells under physiological conditions to an RNA molecule which comprises a nucleic acid sequence as set forth in SEQ ID NO:2, a polynucleotide as set forth by SEQ ID NO:1, a polynucleotide which comprises at least 10 consecutive nucleotides of the nucleic acid sequence set forth in SEQ ID NO:2, a polynucleotide hybridizable in cells under physiological conditions to an RNA molecule which comprises a nucleic acid sequence as set forth in SEQ ID NO:21 and/or 22, a polynucleotide as set forth by SEQ ID NO:2, a polynucleotide which comprises at least 25 consecutive nucleotides of the nucleic acid sequence set forth in SEQ ID NO:13, a polynucleotide as set forth by SEQ ID NO:13, a polynucleotide which comprises at least 20 consecutive nucleotides of SEQ ID NO:13 and/or at least 10 consecutive nucleotides of SEQ ID NO:1, a polynucleotide as set forth by SEQ ID NO:12 or a functional homolog thereof, a polynucleotide as set forth by SEQ ID: 19 or a functional homolog thereof, a polynucleotide as set forth by SEQ ID NO:23 and a polynucleotide as set forth by SEQ ID NO: 24.

### Claim 18 (depends on 15)

The method of claim 15, wherein said miRNA is set forth by the sequence selected from the group consisting of SEQ ID NOs: 54, 93, 94, 98, 99, 100, 21 and 22.

### Claim 19

A method of diagnosing a pathology associated with abnormal function of a miRNA component of an AChE-associated biological pathway in a subject, the method comprising obtaining a biological sample from the subject and determining a level of the miRNA in cells of said biological sample, wherein a level of the miRNA above or below a predetermined threshold or range is indicative of a presence of a pathology associated with abnormal function of the miRNA.

### Claim 20 (depends on 19)

The method of claim 19, wherein said miRNA component is set forth by SEQ NO:21 or SEQ ID NO: 22.

### Claim 21 (depends on 19)

The method of claim 19, wherein said miRNA component is set forth by a sequence selected from the group consisting of SEQ ID NOs: 54, 93, 94, 98, 99 and 100.

### Claim 22 (depends on 19)

The method of claim 19, wherein said determining is effected using an oligonucleotide.

### Claim 23 (depends on 22)

The method of claim 22, wherein said oligonucleotide specifically hybridizable with said miRNA under stringent hybridization conditions.

### Claim 24 (depends on 22)

The method of claim 22, wherein said oligonucleotide is capable of specifically hybridizing with a polynucleotide having a nucleic acid sequence as set forth by SEQ ID NO:21 and/or 22 under stringent hybridization conditions.

### Claim 25 (depends on 19)

The method of claim 19, wherein said determining is effected using at least one oligonucleotide capable of specifically amplifying a polynucleotide having a nucleic acid sequence as set forth in SEQ ID NO:21 and/or 22.

### Claim 26 (depends on 19)

The method of claim 19, wherein said biological sample is selected from the group consisting of blood, bone marrow, spinal fluid and cord blood.

### Claim 27

An isolated polynucleotide comprising a nucleic acid sequence which comprises at least 10 consecutive nucleotides of the nucleotide sequence set forth in SEQ ID NO:1, with the proviso that isolated polynucleotide is not identical to the sequence set forth in SEQ ID NO:1.

### Claim 28

An isolated polynucleotide comprising a nucleic acid sequence of 10-50 bases and capable of hybridizing in cells under physiological conditions with an RNA molecule which comprises a nucleotide sequence as set forth in SEQ ID NO:2, with the proviso that isolated polynucleotide is not identical to the sequence set forth in SEQ ID NO:1.

### Claim 29 (depends on 27)

The isolated polynucleotide of claim 27, wherein the polynucleotide is a modified polynucleotide.

### Claim 30 (depends on 27)

The isolated polypeptide of claim 27, as set forth by SEQ ID NO:23 or 24.

### Claim 31 (depends on 27)

A pharmaceutical composition comprising as an active ingredient the polynucleotide of claim 27 and a pharmaceutically acceptable carrier.

### Claim 32

An isolated polynucleotide comprising a nucleic acid sequence which comprises at least 10 consecutive nucleotides from the nucleotide sequence set forth by SEQ ID NO:2.

### Claim 33

An isolated polynucleotide comprising a nucleic acid sequence of 10-50 bases and capable of hybridizing in cells under physiological conditions with an RNA molecule which comprises a nucleotide sequence as set forth in SEQ ID NO:1.

### Claim 34 (depends on 33)

The isolated polynucleotide of claim 33, wherein said nucleic acid sequence is as set forth in SEQ ID NO:2.

### Claim 35 (depends on 33)

The isolated polynucleotide of claim 33, wherein the polynucleotide is a modified polynucleotide.

### Claim 36 (depends on 35)

The isolated polypeptide of claim 35, wherein said modified polynucleotide is set forth by SEQ ID NO:23 or 24.

### Claim 37 (depends on 33)

A pharmaceutical composition comprising, as an active ingredient the polynucleotide of claim 33 and a pharmaceutically acceptable carrier.

### Claim 38

An isolated polynucleotide comprising a nucleic acid sequence which comprises at least 20 consecutive nucleotides from the nucleotide sequence set forth in SEQ ID NO:13, with the proviso that isolated polynucleotide is not identical to the sequence set forth in SEQ ID NO:13.

### Claim 39 (depends on 38)

The isolated polynucleotide of claim 38, wherein the polynucleotide is a modified polynucleotide.

### Claim 40 (depends on 39)

The isolated polypeptide of claim 39, as set forth by SEQ ID NO:23 or 24.

### Claim 41 (depends on 38)

A pharmaceutical composition comprising, as an active ingredient, the polynucleotide of claim 38 and a pharmaceutically acceptable carrier.