Premature-termination-codons readthrough compounds

A method of screening for readthrough compounds having the ability to read through premature termination codons (PTCs) in RNA, comprising:

• providing a plasmid template to a reaction well having a test compound to cause a coupled protein transcription/translation (PTT) reaction to occur to generate a PTT reaction product, the plasmid template comprising a fragment of ATM gene that contains a PTC mutation, which fragment being flanked by a sequence that initiates transcription of:
• a) a myc epitope,
• b) the ATM fragment, and
• c) a V5 epitope;
• adding the PTT reaction product to high throughput reading trays, which are coated with an antibody to the myc epitope to capture a protein fragment of the fragment of ATM gene,
• adding an antibody to the V5 epitope (V5 antibody) to wells in the reading trays,
• detecting the attachment of the V5 antibody to proteins in the PTT product, and
• identifying the test compound as a readthrough compound if the attachment of the V5 antibody to proteins in the PTT product occurs.

A compound having the ability to read through premature termination codons in RNA having a structure of

• or a pharmaceutically acceptable salt thereof,
• wherein for formula (III):
• X is O or S;
• Z is O, S, or NR′ where R′ is hydrogen or a C1-C6 group;
• and
• R is an ortho, meta, or para group and is selected from hydrogen, C1-C6 group, hydroxyl group, alkoxy, halo, trifluoromethyl, amino, alkylamino, thiol, alkylthio, sulfonyl, formyl, or carboxyl group;
• wherein for formula (IV) and (V):
• R1 is ortho, meta, or para to R2 and is a hydrogen, C1-C6 group, hydroxyl group, alkoxy, halo, amino, alkylamino, thiol, alkylthio, sulfonyl, nitro, formyl, or carboxyl group, and
• R2 is a hydroxyl, methoxy, ethoxy, halo, amino, alkylamino, thiol, alkylthio, sulfonyl, nitro, or carboxyl group;
• and
• R3 is an ortho, meta, or para group and is a hydrogen, a C1-C6 group, hydroxyl, alkoxy, halo, amino, alkylamino, thiol, alkylthio, sulfonyl, nitro, or carboxyl group;
• wherein for formula (VI) and (VII):
• X is O or S;
• R1 is ortho, meta, or para to R2 and is a hydrogen, C1-C6 group, hydroxyl, alkoxy, halo, amino, alkylamino, thiol, alkylthio, sulfonyl, nitro, formyl, or carboxyl group;
• R2 is a hydroxyl, methoxy, ethoxy, halo, amino, alkylamino, thiol, alkylthio, sulfonyl, nitro, or carboxyl group;
• and
• R3 is ortho, meta, or para group and is a hydrogen, a C1-C6 group, hydroxyl, alkoxy, halo, amino, alkylamino, thiol, alkylthio, sulfonyl, nitro, or carboxyl group;
• wherein for formula (VIA) and (VIIA)::
• R1 is hydrogen or a C1-C6, hydroxyl group, alkoxy, halo, amino, alkylamino, thiol, alkylthio, sulfonyl, nitro, formyl, or carboxyl group,
• R2 is a hydroxyl, methoxy, ethoxy, halo, amino, alkylamino, thiol, alkylthio, sulfonyl, nitro, or carboxyl group;
• and
• R3 is ortho, meta, or para and is a hydrogen, a C1-C6 group, hydroxyl, alkoxy, halo, amino, alkylamino, thiol, alkylthio, sulfonyl, nitro, or carboxyl group;
• and
• provided that for the compound of formula VIA, R1, R2 and R3 are not simultaneously halo;
• provided that the compound of formula VIA is not 3-(((5-(2-chlorophenyl)furan-2-yl)methyl)amino)-4-methoxybenzoic acid, 4-chloro-3-(((5-(2-chlorophenyl)furan-2-yl)methyl)amino)benzoic acid, 4-chloro-3-(((5-(2-fluorophenyl)furan-2-yl)methyl)amino)benzoic acid or 3-(((5-(2-fluorophenyl)furan-2-yl)methyl)amino)-4-methoxybenzoic acid;
• and provided that the compound of formula VIIA is not N-((5-(2-chlorophenyl)furan-2-yl)methylene)-2-methoxy-5-nitroaniline or 4-chloro-2-(((5-(2-nitrophenyl)furan-2-yl)methylene)amino)phenol.

A compound selected from the group consisting of

• (5Z)-2-imino-5-[[5-(2-nitrophenyl)-2-furyl]methylene]thiazolidin-4-one
• (5Z)-5-[[5-(2-nitrophenyl)-2-furyl]methylene]-2-thioxo-thiazolidin-4-one
• 4-tert-butyl-2-[(3-nitrophenyl)methyleneamino]phenol
• 2-amino-4-tert-butyl-phenol
• 2-[(3-nitrophenyl)methyleneamino]phenol
• 2-(benzylideneamino)-4-tert-butyl-phenol
• 4-tert-butyl-2-[(3-nitrophenyl)methylamino]phenol.

A method of treating or ameliorating a medical condition associated with premature termination codons (PTCs) in RNA, comprising administering to a subject a compound according to formula (III), (IV), (V), (VI), or (VII) or a composition comprising the compound of formula (III), (IV), (V), (VI), or (VII) and a pharmaceutically acceptable carrier;

• wherein
• wherein for formula (III):
• X is O or S;
• Z is O, S, or NR′ where R′ is hydrogen or a C1-C6 group;
• and
• R is an ortho, meta, or para group and is selected from hydrogen, C1-C6 group, hydroxyl group, alkoxy, halo, trifluoromethyl, amino, alkylamino, thiol, alkylthio, sulfonyl, formyl, or carboxyl group;
• wherein for formula (IV) and (V):
• R1 is ortho, meta, or para to R2 and is a hydrogen, C1-C6 group, hydroxyl group, alkoxy, halo, amino, alkylamino, thiol, alkylthio, sulfonyl, nitro, formyl, or carboxyl group, and
• R2 is a hydroxyl, methoxy, ethoxy, halo, amino, alkylamino, thiol, alkylthio, sulfonyl, nitro, or carboxyl group;
• and
• R3 is an ortho, meta, or para group and is a hydrogen, a C1-C6 group, hydroxyl, alkoxy, halo, amino, alkylamino, thiol, alkylthio, sulfonyl, nitro, or carboxyl group;
• wherein for formula (VI) and (VII):
• X is O or S;
• R1 is ortho, meta, or para to R2 and is a hydrogen, C1-C6 group, hydroxyl, alkoxy, halo, amino, alkylamino, thiol, alkylthio, sulfonyl, nitro, formyl, or carboxyl group;
• R2 is a hydroxyl, methoxy, ethoxy, halo, amino, alkylamino, thiol, alkylthio, sulfonyl, nitro, or carboxyl group;
• and
• R3 is ortho, meta, or para group and is a hydrogen, a C1-C6 group, hydroxyl, alkoxy, halo, amino, alkylamino, thiol, alkylthio, sulfonyl, nitro, or carboxyl group.

A compound having the ability to read through premature termination codons (PTCs) in RNA, a pharmaceutically acceptable salt thereof, having a structure of formula (VIII):

• wherein:
• X is O;
• R3 is an ortho, meta, or para group and is a hydrogen, halo, or nitro;
• and
• R4 is hydrogen, —C(O)H, halo, or a C1-C6 group.