# US 2014/0155461 A1 · Molecular targets and compounds, and methods to identify the same, useful in the treatment of bone and joint degenerative diseases

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- Number: US 2014/0155461 A1
- Document ID: US20140155461A1
- Status: Published application
- Kind: A1
- Application number: 14/106,951
- Filed: 2013-12-16
- Published: 2014-06-05
- Earliest claimed: 2007-06-20
- Inventors: Reginald Christophe Xavier Brys; Nick Ernest Rene Vandeghinste
- Web page: https://patentgrep.com/patent/US20140155461A1

## CPC

- G01N 33/5308: {for analytes not provided for elsewhere, e.g. nucleic acids, uric acid, worms, mites}
- A61K 31/7088: Compounds having three or more nucleosides or nucleotides
- A61K 31/713: Double-stranded nucleic acids or oligonucleotides
- A61K 45/06: Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
- A61P 19/00: Drugs for skeletal disorders
- A61P 19/02: for joint disorders, e.g. arthritis, arthrosis
- A61P 19/08: for bone diseases, e.g. rachitism, Paget's disease
- A61P 19/10: for osteoporosis
- A61P 29/00: Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
- C12N 15/111: {General methods applicable to biologically active non-coding nucleic acids}
- C12Y 111/01006: Catalase (1.11.1.6)
- G01N 33/6887: {from muscle, cartilage or connective tissue}
- C12N 2310/111: spanning the whole gene, or a large part of it
- C12N 2310/14: interfering nucleic acids [NA]
- C12N 2310/53: partially self-complementary or closed
- C12N 2320/12: in functional genomics, i.e. for the determination of gene function
- G01N 2500/04: Screening involving studying the effect of compounds C directly on molecule A (e.g. C are potential ligands for a receptor A, or potential substrates for an enzyme A)
- G01N 2800/102: Arthritis; Rheumatoid arthritis, i.e. inflammation of peripheral joints
- G01N 2800/105: Osteoarthritis, e.g. cartilage alteration, hypertrophy of bone
- G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q)
- 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)
- A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- C12Y: ENZYMES

## Abstract

The present invention relates to methods for identifying agents capable of inhibiting the expression or activity of proteins involved in the processes modulating osteoclastogenesis, which inhibition is useful in the prevention and/or treatment of bone and joint degenerative diseases and diseases involving aberrant activity or differentiation of osteoclasts. In particular, the present invention provides methods for identifying agents for use in the prevention and/or treatment of rheumatoid arthritis.

## Claims

### Claim 1

A method for identifying a compound that inhibits bone resorption, comprising:
(a) contacting a compound with a polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 41-69 and 80, and fragments thereof; and
(b) measuring a compound-polypeptide property related to bone resorption.

### Claim 2 (depends on 1)

The method according to claim 1, wherein said polypeptide is in an in vitro cell-free preparation.

### Claim 3 (depends on 1)

The method according to claim 1, wherein said polypeptide is present in a mammalian cell.

### Claim 4 (depends on 2)

The method of claim 2, wherein said property is a binding affinity of said compound to said polypeptide.

### Claim 5 (depends on 4)

The method of claim 4, which additionally comprises the steps of:
c) contacting a population of mammalian cells expressing said polypeptide with the compound that exhibits a binding affinity of at least 10 micromolar; and
d) identifying a compound that inhibits bone resorption.

### Claim 6 (depends on 1)

The method of claim 1, wherein said property is upregulation of a biological pathway producing a biochemical marker indicative of the inhibition of bone resorption.

### Claim 7 (depends on 6)

The method of claim 6 wherein said indicator is osteoprotegerin.

### Claim 8 (depends on 1)

The method of claim 1, wherein said property is the activity of said polypeptide.

### Claim 9 (depends on 1)

The method of claim 1, wherein said property is the expression of said polypeptide.

### Claim 10 (depends on 8, 9)

The method according to claim 8 or 9, which additionally comprises the steps of:
c) contacting a population of mammalian cells expressing said polypeptide with the compound that significantly inhibits the expression or activity of the polypeptide; and
d) identifying the compound that inhibits bone resorption.

### Claim 11 (depends on 1)

The method according to claim 1, which additionally comprises the step of comparing the compound to be tested to a control.

### Claim 12 (depends on 11)

The method according to claim 11, wherein said control is where the polypeptide has not been contacted with said compound.

### Claim 13 (depends on 5, 10)

The method according to claim 5 or 10, which additionally comprises the step of comparing the compound to a control, wherein said control is a population of mammalian cells that does not express said polypeptide.

### Claim 14 (depends on 1)

The method according to claim 1, wherein said compound is selected from the group consisting of compounds of a commercially available screening library and compounds having binding affinity for a polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 41-69 and 80.

### Claim 15 (depends on 1)

The method according to claim 1, wherein said compound is a peptide in a phage display library or an antibody fragment library.

### Claim 16

An agent effective in inhibiting bone resorption, selected from the group consisting of an antisense polynucleotide, a ribozyme, and a small interfering RNA (siRNA), wherein said agent comprises a nucleic acid sequence complementary to, or engineered from, a naturally-occurring polynucleotide sequence of about 17 to about 30 contiguous nucleotides of a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1-29 and 40.

### Claim 17 (depends on 16)

The agent according to claim 16, wherein a vector in a mammalian cell expresses said agent.

### Claim 18 (depends on 16)

The agent according to claim 16, which is effective in inducing osteoprotegerin (OPG) expression in the OPG assay.

### Claim 19 (depends on 17)

The agent according to claim 17, wherein said vector is an adenoviral, retroviral, adeno-associated viral, lentiviral, a herpes simplex viral or a sendaiviral vector.

### Claim 20 (depends on 16)

The agent according to claim 16, wherein said antisense polynucleotide and said siRNA comprise an antisense strand of 17-25 nucleotides complementary to a sense strand, wherein said sense strand is selected from 17-25 continuous nucleotides of a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1-29 and 40.

### Claim 21 (depends on 20)

The agent according to claim 20, wherein said siRNA further comprises said sense strand.

### Claim 22 (depends on 21)

The agent according to claim 21, wherein said sense strand is selected from the group consisting of SEQ ID NO: 81-97 and 107.

### Claim 23 (depends on 20)

The agent according to claim 20, wherein said siRNA further comprises a loop region connecting said sense and said antisense strand.

### Claim 24 (depends on 23)

The agent according to claim 23, wherein said loop region comprises a nucleic acid sequence selected from the group consisting of UUGCUAUA or GUUUGCUAUAAC (SEQ ID NO: 108).

### Claim 25 (depends on 16)

The agent according to claim 16, wherein said agent is an antisense polynucleotide, ribozyme, or siRNA comprising a nucleic acid sequence complementary to a nucleic acid sequence selected from the group consisting of SEQ ID NO: 81-97 and 107.

### Claim 26 (depends on 16)

A bone resorption inhibiting pharmaceutical composition comprising a therapeutically effective amount of an agent according to claim 16 in admixture with a pharmaceutically acceptable carrier.

### Claim 27 (depends on 26)

A method for treatment and/or prevention of a disease involving an imbalance in bone metabolism in a subject suffering from or susceptible to the disease comprising administering to the subject the pharmaceutical composition of claim 26.

### Claim 28 (depends on 27)

The method according to claim 27 wherein the disease is a joint degenerative disease.

### Claim 29 (depends on 28)

The method according to claim 28, wherein the disease is rheumatoid arthritis.

### Claim 30 (depends on 16)

A method for treatment and/or prevention of a disease involving abnormal bone resorption in a subject comprising administering to the subject the agent of claim 16.

### Claim 31 (depends on 30)

The method according to claim 30, wherein the disease is selected from the group consisting of joint degenerative and inflammation diseases.

### Claim 32 (depends on 30)

The method according to claim 30, wherein the disease is rheumatoid arthritis.

### Claim 33 (depends on 16)

A method for treatment or prevention of a condition characterized by abnormal osteoprotegrin (OPG) expression and/or activity in a subject comprising administering to the subject the agent of claim 16.

### Claim 34 (depends on 27, 30, 33)

The method of any of claim 27, 30 or 33 wherein the treatment and/or prevention additionally comprises administering said pharmaceutical composition or said OPG inducing agent in combination with a disease-modifying anti-rheumatic drug (DMARD) or an anti-inflammatory compound.

### Claim 35 (depends on 34)

The method according to claim 34, wherein said DMARD is selected from the group consisting of Infliximab, Etanercept, Adalimumab, Rituximab, CTLA4-Ig methotrexate, leflunomide and sulfasalazine.

### Claim 36 (depends on 34)

The method according to claim 34, wherein said anti-inflammatory agent is selected from the group consisting of corticosteroids or non-steroidal anti-inflammatory agents.

### Claim 37

A method for diagnosing a pathological condition involving abnormal bone resorption or a susceptibility to the condition in a subject, comprising determining a first amount of polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 41-67 and 80 present in a biological sample obtained from said subject, and comparing said first amount with the ranges of amounts of the polypeptide determined in a population of healthy subjects, wherein an increase of the amount of polypeptide in said biological sample compared to the range of amounts determined for healthy subjects is indicative of the presence of the pathological condition.