# US 9,433,610 B2 · Compounds that enhance Atoh1 expression

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- Number: US 9,433,610 B2
- Document ID: US9433610B2
- Status: Granted
- Kind: B2
- Application number: 14/511,911
- Filed: 2014-10-10
- Published: 2015-06-11
- Granted: 2016-09-06
- Earliest claimed: 2008-02-07
- Assignees: Massachusetts Eye and Ear Infirmary; The Brigham and Women's Hospital, Inc.
- Applicants: Massachusetts Eye & Ear Infirmary; The Brigham & Women's Hospital, Inc.
- Inventors: Albert Edge; Sang-Jun Jeon; Kathleen Seyb; Marcie Glicksman; Lixin Qiao; Gregory D. Cuny
- Web page: https://patentgrep.com/patent/US9433610B2

## CPC

- A61K 31/5377: not condensed and containing further heterocyclic rings, e.g. timolol
- A61K 9/0046: {Ear}
- A61K 31/341: not condensed with another ring, e.g. ranitidine, furosemide, bufetolol, muscarine
- A61K 31/343: condensed with a carbocyclic ring, e.g. coumaran, bufuralol, befunolol, clobenfurol, amiodarone
- A61K 31/352: condensed with carbocyclic rings, e.g. methantheline {(cannabinoids A61K31/658)}
- A61K 31/353: 3,4-Dihydrobenzopyrans, e.g. chroman, catechin
- A61K 31/381: having five-membered rings
- A61K 31/404: Indoles, e.g. pindolol
- A61K 31/415: 1,2-Diazoles
- A61K 31/4155: non condensed and containing further heterocyclic rings
- A61K 31/4184: condensed with carbocyclic rings, e.g. benzimidazoles
- A61K 31/4192: 1,2,3-Triazoles
- A61K 31/423: condensed with carbocyclic rings
- A61K 31/4245: Oxadiazoles
- A61K 31/426: 1,3-Thiazoles
- A61K 31/428: condensed with carbocyclic rings
- A61K 31/4365: the heterocyclic ring system having sulfur as a ring hetero atom, e.g. ticlopidine
- A61K 31/437: the heterocyclic ring system containing a five-membered ring having nitrogen as a ring hetero atom, e.g. indolizine, beta-carboline
- A61K 31/44: Non condensed pyridines; Hydrogenated derivatives thereof
- A61K 31/4436: containing a heterocyclic ring having sulfur as a ring hetero atom
- A61K 31/4439: containing a five-membered ring with nitrogen as a ring hetero atom, e.g. omeprazole (nicotine A61K31/465)
- A61K 31/496: Non-condensed piperazines containing further heterocyclic rings, e.g. rifampin, thiothixene or sparfloxacin
- A61K 31/498: Pyrazines or piperazines ortho- and peri-condensed with carbocyclic ring systems, e.g. quinoxaline, phenazine
- A61K 31/517: ortho- or peri-condensed with carbocyclic ring systems, e.g. quinazoline, perimidine
- A61K 31/519: ortho- or peri-condensed with heterocyclic rings
- A61K 35/28: Bone marrow; Haematopoietic stem cells; Mesenchymal stem cells of any origin, e.g. adipose-derived stem cells
- A61K 35/545: Embryonic stem cells; Pluripotent stem cells; Induced pluripotent stem cells; Uncharacterised stem cells
- A61P 19/00: Drugs for skeletal disorders
- A61P 19/02: for joint disorders, e.g. arthritis, arthrosis
- A61P 25/00: Drugs for disorders of the nervous system
- A61P 27/00: Drugs for disorders of the senses
- A61P 27/02: Ophthalmic agents
- A61P 27/16: Otologicals
- A61P 35/00: Antineoplastic agents
- A61P 43/00: Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
- 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

## Abstract

This invention generally provides compounds, pharmaceutical compositions, and methods for their use, which include methods that result in increased expression in an Atoh1 gene (e.g., Hath1) in a biological cell. More specifically, the invention relates to the treatment of diseases and/or disorders that would benefit from increased Atoh1 expression, e.g., a hearing impairment or imbalance disorder associated with a loss of auditory hair cells, or a disorder associated with abnormal cellular proliferation.

## Claims

### Claim 1

A method of treating a hearing impairment or imbalance disorder associated with loss of auditory hair cells in a subject, the method comprising selecting a subject in need of treatment, and administering to the subject a therapeutically effective amount of a pharmaceutical composition comprising a compound of the following formula:
or a pharmaceutically acceptable salt thereof, wherein:
X is O or S;
each of R143, R144, R145, and R146 is, independently selected from H, halo, OH, CN, NO2, C1-C3 alkyl, C1-C3 haloalkyl, C1-C3 alkoxy, C1-C3 haloalkoxy; and —NHC(O)(C1-C3 alkyl);
R147 is NReRf, wherein one of Re and Rf is hydrogen or C1-C3 alkyl; and the other of Re and Rf is:
(i) —C(O)Rg; wherein Rg is C6-C10 aryl or heteroaryl including 5-10 atoms, each of which is optionally substituted with from 1-5 Rh; or
(ii) C1-C3 alkyl;
or
R147 is C6-C10 aryl or heteroaryl including 5-10 atoms, each of which is optionally substituted with from 1-5 Rh;
or
R147 is —SCH2Ri, wherein Ri is:
(i) C6-C10 aryl or heteroaryl including 5-10 atoms, each of which is optionally substituted with from 1-5 Rh; or
(ii) —C(O)NReRf, wherein one of Re and Rf is hydrogen or C1-C3 alkyl; and the other of Re and Rf is —C(O)Rg; wherein Rg is C6-C10 aryl or heteroaryl including 5-10 atoms, each of which is optionally substituted with from 1-5 Rh; and
Rh at each occurrence is, independently:
(i) halo; NH2; NH(C1-C3 alkyl); N(C1-C3 alkyl)2; hydroxy; C1-C6 alkoxy or C1-C6 haloalkoxy; nitro; or cyano; or
(ii) C1-C6 alkyl or C1-C6 haloalkyl;
and a pharmaceutically acceptable carrier.

### Claim 2 (depends on 1)

The method of claim 1, wherein X is O.

### Claim 3 (depends on 1)

The method of claim 1, wherein X is S.

### Claim 4 (depends on 1)

The method of claim 1, wherein each of R143, R144, R145, and R146 is hydrogen.

### Claim 5 (depends on 1)

The method of claim 1, wherein one of R143, R144, R145, and R146 is halo, OH, CN, NO2, C1-C3 alkyl, C1-C3 haloalkyl, C1-C3 alkoxy, C1-C3 haloalkoxy; or —NHC(O)(C1-C3 alkyl); and the others are hydrogen.

### Claim 6 (depends on 1)

The method of claim 1, wherein R147 is C6-C10 aryl optionally substituted with from 1-5 Rh.

### Claim 7 (depends on 1)

The method of claim 1, wherein the compound is selected from the group consisting of:
N-(6-bromobenzo[d]thiazol-2-yl)thiophene-2-carboxamide;
N-(benzo[d]thiazol-2-yl)-1-methyl-1H-pyrazole-5-carboxamide;
2-(4-fluorobenzylthio)benzo[d]thiazole;
5-chloro-N-methylbenzo[d]thiazol-2-amine;
N-(6-acetamidobenzo[d]thiazol-2-yl)furan-2-carboxamide;
N-(6-fluorobenzo[d]thiazol-2-yl)-3-methoxybenzamide;
2-(benzo[d]oxazol-2-ylthio)-N-(2-chlorophenyl)acetamide;
5-chloro-2-phenylbenzo[d]oxazole;
5-methyl-2-m-tolylbenzo[d]oxazole,
or a pharmaceutically acceptable salt thereof.

### Claim 8 (depends on 1)

The method of claim 1, wherein the subject in need of treatment has, or is at risk of developing, a hearing impairment or imbalance disorder associated with loss of auditory hair cells, and wherein the pharmaceutical composition is administered locally to the inner ear of the subject and increases the number of cells in the subject's inner ear that have characteristics of auditory hair cells.

### Claim 9 (depends on 8)

The method of claim 8, wherein the pharmaceutical composition is administered by injection into one or more of the luminae of the cochlea, into the auditory nerve trunk in the internal auditory meatus, or into the scala tympani.

### Claim 10 (depends on 8)

The method of claim 8, wherein the auditory hair cells are outer or inner auditory hair cells.

### Claim 11

A method of treating a gastrointestinal cancer in a subject, the method comprising selecting a subject in need of treatment, and administering to the subject a therapeutically effective amount of a pharmaceutical composition comprising a compound of the following formula:
or a pharmaceutically acceptable salt thereof, wherein:
X is O or S;
each of R143, R144, R145, and R146 is, independently selected from H, halo, OH, CN, NO2, C1-C3 alkyl, C1-C3 haloalkyl, C1-C3 alkoxy, C1-C3 haloalkoxy; and —NHC(O)(C1-C3 alkyl);
R147 is NReRf, wherein one of Re and Rf is hydrogen or C1-C3 alkyl; and the other of Re and Rf is:
(i) —C(O)Rg; wherein Rg is C6-C10 aryl or heteroaryl including 5-10 atoms, each of which is optionally substituted with from 1-5 Rh; or
(ii) C1-C3 alkyl;
or
R147 is C6-C10 aryl or heteroaryl including 5-10 atoms, each of which is optionally substituted with from 1-5 Rh;
or
R147 is —SCH2Ri, wherein Ri is:
(i) C6-C10 aryl or heteroaryl including 5-10 atoms, each of which is optionally substituted with from 1-5 Rh; or
(ii) —C(O)NReRf, wherein one of Re and Rf is hydrogen or C1-C3 alkyl; and the other of Re and Rf is —C(O)Rg; wherein Rg is C6-C10 aryl or heteroaryl including 5-10 atoms, each of which is optionally substituted with from 1-5 Rh; and
Rh at each occurrence is, independently:
(i) halo; NH2; NH(C1-C3 alkyl); N(C1-C3 alkyl)2; hydroxy; C1-C6 alkoxy or C1-C6 haloalkoxy; nitro; or cyano; or
(ii) C1-C6 alkyl or C1-C6 haloalkyl;
and a pharmaceutically acceptable carrier,
wherein the pharmaceutical composition is administered locally to the proliferating cells.

### Claim 12 (depends on 11)

The method of claim 11, wherein the gastrointestinal cancer is selected from the group consisting of cancer of the esophagus, gallbladder, liver, pancreas, stomach, small intestine, large intestine, colon, and rectum.

### Claim 13

A method of treating cerebellar granule neuron deficiencies, joint disease, or osteoarthritis in a subject, the method comprising selecting a subject in need of treatment, and administering to the subject a therapeutically effective amount of a pharmaceutical composition comprising a compound of the following formula:
or a pharmaceutically acceptable salt thereof, wherein:
X is O or S;
each of R143, R144, R145, and R146 is, independently selected from H, halo, OH, CN, NO2, C1-C3 alkyl, C1-C3 haloalkyl, C1-C3 alkoxy, C1-C3 haloalkoxy; and —NHC(O)(C1-C3 alkyl);
R147 is NReRf, wherein one of Re and Rf is hydrogen or C1-C3 alkyl; and the other of Re and Rf is:
(i) —C(O)Rg; wherein Rg is C6-C10 aryl or heteroaryl including 5-10 atoms, each of which is optionally substituted with from 1-5 Rh; or
(ii) C1-C3 alkyl;
or
R147 is C6-C10 aryl or heteroaryl including 5-10 atoms, each of which is optionally substituted with from 1-5 Rh;
or
R147 is —SCH2Ri, wherein Ri is:
(i) C6-C10 aryl or heteroaryl including 5-10 atoms, each of which is optionally substituted with from 1-5 Rh; or
(ii) —C(O)NReRf, wherein one of Re and Rf is hydrogen or C1-C3 alkyl; and the other of Re and Rf is —C(O)Rg; wherein Rg is C6-C10 aryl or heteroaryl including 5-10 atoms, each of which is optionally substituted with from 1-5 Rh; and
Rh at each occurrence is, independently:
(i) halo; NH2; NH(C1-C3 alkyl); N(C1-C3 alkyl)2; hydroxy; C1-C6 alkoxy or C1-C6 haloalkoxy; nitro; or cyano; or
(ii) C1-C6 alkyl or C1-C6 haloalkyl;
and a pharmaceutically acceptable carrier.

### Claim 14

A method of treating a subject who has a hearing impairment or imbalance disorder associated with a loss of auditory hair cells, the method comprising:
selecting a subject in need of treatment;
obtaining a population of cells capable of differentiating into auditory hair cells;
contacting the population of cells in vitro with an effective amount of at least one compound of the following formula:
or a pharmaceutically acceptable salt thereof, wherein:
X is O or S;
each of R143, R144, R145, and R146 is, independently selected from H, halo, OH, CN, NO2, C1-C3 alkyl, C1-C3 haloalkyl, C1-C3 alkoxy, C1-C3 haloalkoxy, and —NHC(O)(C1-C3 alkyl);
R147 is NReRf, wherein one of Re and Rf is hydrogen or C1-C3 alkyl, and the other of Re and Rf is:
(i) —C(O)Rg; wherein Rg is C6-C10 aryl or heteroaryl including 5-10 atoms, each of which is optionally substituted with from 1-5 Rh, or
(ii) C1-C3 alkyl;
or
R147 is C6-C10 aryl or heteroaryl including 5-10 atoms, each of which is optionally substituted with from 1-5 Rh;
or
R147 is —SCH2Ri, wherein Ri is:
(i) C6-C10 aryl or heteroaryl including 5-10 atoms, each of which is optionally substituted with from 1-5 Rh, or
(ii) —C(O)NReRf, wherein one of Re and Rf is hydrogen or C1-C3 alkyl, and the other of Re and Rf is —C(O)Rg; wherein Rg is C6-C10 aryl or heteroaryl including 5-10 atoms, each of which is optionally substituted with from 1-5 Rh, and
Rh at each occurrence is, independently:
(i) halo; NH2; NH(C1-C3 alkyl); N(C1-C3 alkyl)2; hydroxy; C1-C6 alkoxy or C1-C6 haloalkoxy; nitro; or cyan; or
(ii) C1-C6 alkyl or C1-C6 haloalkyl,
for a time sufficient to increase the number cells in the population that have characteristics of a differentiated auditory hair cell; and
administering the population of cells, or a subset thereof, to the subject's ear, thereby treating the subject.

### Claim 15 (depends on 14)

The method of claim 14, wherein the population of cells capable of differentiating into auditory hair cells includes cells selected from the group consisting of stem cells, progenitor cells, support cells, Deiters' cells, pillar cells, inner phalangeal cells, tectal cells, Hensen's cells, and germ cells.

### Claim 16 (depends on 15)

The method of claim 15, wherein the stem cells are adult stem cells.

### Claim 17 (depends on 16)

The method of claim 16, wherein the adult stem cells are derived from inner ear, bone marrow, mesenchyme, skin, fat, liver, muscle, or blood.

### Claim 18 (depends on 15)

The method of claim 15, wherein the stem cells are embryonic stem cells or stem cells obtained from a placenta or umbilical cord.

### Claim 19 (depends on 15)

The method of claim 15, wherein the progenitor cells are derived from the inner ear, bone marrow, mesenchyme, skin, fat, liver, muscle, or blood.

### Claim 20 (depends on 14)

The method of claim 14, wherein administering the population of cells comprises (a) injecting the cells into the luminae of the cochlea, into the auditory nerve trunk in the internal auditory meatus, or into the scala tympani; or (b) implanting the cells within a cochlear implant.

### Claim 21 (depends on 14)

The method of claim 14, further comprising administering to the subject a pharmaceutical composition comprising a compound of the following formula:
or a pharmaceutically acceptable salt thereof, wherein:
X is O or S;
each of R143, R144, R145, and R146 is, independently selected from H, halo, OH, CN, NO2, C1-C3 alkyl, C1-C3 haloalkyl, C1-C3 alkoxy, C1-C3 haloalkoxy; and —NHC(O)(C1-C3 alkyl);
R147 is NReRf, wherein one of Re and Rf is hydrogen or C1-C3 alkyl; and the other of Re and Rf is:
(i) —C(O)Rg; wherein Rg is C6-C10 aryl or heteroaryl including 5-10 atoms, each of which is optionally substituted with from 1-5 Rh; or
(ii) C1-C3 alkyl;
or
R147 is C6-C10 aryl or heteroaryl including 5-10 atoms, each of which is optionally substituted with from 1-5 Rh;
or
R147 is —SCH2Ri, wherein Ri is:
(i) C6-C10 aryl or heteroaryl including 5-10 atoms, each of which is optionally substituted with from 1-5 Rh; or
(ii) —C(O)NReRf, wherein one of Re and Rf is hydrogen or C1-C3 alkyl; and the other of Re and Rf is —C(O)Rg; wherein Rg is C6-C10 aryl or heteroaryl including 5-10 atoms, each of which is optionally substituted with from 1-5 Rh; and
Rh at each occurrence is, independently:
(i) halo; NH2; NH(C1-C3 alkyl); N(C1-C3 alkyl)2; hydroxy; C1-C6 alkoxy or C1-C6 haloalkoxy; nitro; or cyano; or
(ii) C1-C6 alkyl or C1-C6 haloalkyl;
and a pharmaceutically acceptable carrier.

### Claim 22 (depends on 21)

The method of claim 21, wherein the pharmaceutical composition is administered locally to the patient's ear.

### Claim 23 (depends on 22)

The method of claim 22, wherein the pharmaceutical composition is administered by application to the luminae of the cochlea, to the auditory nerve trunk in the internal auditory meatus, to the scala tympani or, if present, into a cochlear implant.

### Claim 24

A method of increasing the number of cells with the characteristics of auditory hair cells in a population of cells in vitro, the method comprising obtaining a population of cells capable of differentiating into auditory hair cells, contacting the population of cells in vitro with an effective amount of at least one compound of the following formula:
or a pharmaceutically acceptable salt thereof, wherein:
X is O or S;
each of R143, R144, R145, and R146 is, independently selected from H, halo, OH, CN, NO2, C1-C3 alkyl, C1-C3 haloalkyl, C1-C3 alkoxy, C1-C3 haloalkoxy; and —NHC(O)(C1-C3 alkyl);
R147 is NReRf, wherein one of Re and Rf is hydrogen or C1-C3 alkyl; and the other of Re and Rf is:
(i) —C(O)Rg; wherein Rg is C6-C10 aryl or heteroaryl including 5-10 atoms, each of which is optionally substituted with from 1-5 Rh; or
(ii) C1-C3 alkyl;
or
R147 is C6-C10 aryl or heteroaryl including 5-10 atoms, each of which is optionally substituted with from 1-5 Rh;
or
R147 is —SCH2Ri, wherein Ri is:
(i) C6-C10 aryl or heteroaryl including 5-10 atoms, each of which is optionally substituted with from 1-5 Rh; or
(ii) —C(O)NReRf, wherein one of Re and Rf is hydrogen or C1-C3 alkyl; and the other of Re and Rf is —C(O)Rg; wherein Rg is C6-C10 aryl or heteroaryl including 5-10 atoms, each of which is optionally substituted with from 1-5 Rh; and
Rh at each occurrence is, independently:
(i) halo; NH2; NH(C1-C3 alkyl); N(C1-C3 alkyl)2; hydroxy; C1-C6 alkoxy or C1-C6 haloalkoxy; nitro; or cyano; or
(ii) C1-C6 alkyl or C1-C6 haloalkyl,
for a time sufficient to increase the number of cells with the characteristics of auditory hair cells in the population of cells.

### Claim 25 (depends on 22)

The method of claim 22, wherein the population of cells capable of differentiating into hair cells comprises cells selected from the group consisting of stem cells, inner ear stem cells, adult stem cells, bone marrow derived stem cells, embryonic stem cells, mesenchymal stem cells, skin stem cells, fat derived stem cells, progenitor cells, inner ear progenitor cells, support cells, Deiters' cells, pillar cells, inner phalangeal cells, tectal cells, Hensen's cells, and germ cells.