Compounds comprising an imidazole core

Abstract

Disclosed are compounds of formula (I), or a pharmaceutically acceptable salt or N-oxide thereof wherein R1, R2, R3, R4, and L1 are as defined herein. Compounds of the invention may be suitable for use in treating diseases / conditions which are associated with PTC mutations. Also disclosed are pharmaceutical compositions comprising the compounds; and the compounds for use in treating conditions or disorders which are associated with PTC mutations in a subject.

Description

Compounds Comprising An Imidazole Core

[0001] This invention relates to novel compounds having pharmacological activity, processes for their preparation, pharmaceutical compositions containing them and their use in the treatment of various disorders.BACKGROUND

[0002] When a ribosome encounters a termination sequence during the process of messenger RNA (mRNA) translation no cognate transfer RNA (tRNA) can bind. This leads to a halt in protein production, release factor (RF) proteins binding to the empty site on the ribosome and conformational changes in the ribosome that lead to the break-up of the ribosomal complex. Although accurate, translational termination is not 100% effective and its efficiency depends on competition between the recognition of the stop codon by eRF1 , and the decoding of the stop codon by a near-cognate tRNA (i.e. a natural suppressor tRNA). The latter case leads to the suppression of translation termination, also called “read-through”, where an amino acid is incorporated in place of the stop (Dabrowski, M. et al., RNA Biology 2015, 12, 950-958).

[0003] If the terminating codon occurs before the full length protein is made then the termination codon is a “premature” termination codon (PTC) and the truncated protein may not be functional. Normal terminating stop codons are usually in a genetic and structural context that reinforces their effectiveness and are usually more resistant to read-through than PTCs arising through truncating mutations (Wangen and Green, 2020 Elife 9:e52611).

[0004] Over 1800 separate inherited genetic disorders have been identified where the causative mutation in a proportion of individuals is a nonsense mutation where an in-frame, premature termination codon (PTC) leads to a loss of gene function (Kellermayer, R. European Journal of Medical Genetics 2006, 49, 445-450). Over 9,600 PTC mutations have been published, representing around 10% of all reported gene mutations (Krawczak, M. et al., Human Mutation 2000, 15, 45-51 , Mort, M. et al., Human Mutation 2008, 29, 1037- 1047).

[0005] Nonsense mutations are involved in the pathology of a range of disorders; from polygenic pathologies such as cancer to monogenic diseases such as cystic fibrosis and Duchenne muscular dystrophy for which a PTC accounts for around 10% or 5-10% of all cases respectively (Keeling, K.M. & Bedwell, D.M., Journal of Molecular Medicine 2002, 80, 367-376, Kellermayer, R.European Journal of Medical Genetics 2006, 49, 445-450).

[0006] Whereas monogenetic diseases arise from either an inherited or de novo germline mutation, resulting in a PTC, in for example cancer, somatic mutations, can give rise to PTC mutations in tumour suppressor genes for example.

[0007] One of the surveillance systems used by the cell to remove abnormal mRNA transcripts that prematurely terminate translation is the non-sense mediated decay (NMD) pathway. It targets mRNA containing a PTC sequence more than ~50 base pairs upstream from the penultimate exon-exon junction (Maquat, L.E., Nature Reviews, Molecular Cell biology 2004, 5, 89-99).

[0008] The combination of premature termination of protein production and NMD of mRNA containing PTCs, can lead to almost total loss of protein production for mutant genes encoding a PTC.

[0009] Mechanisms by which translational read-though of PTC mutations may be achieved include but are not limited to: reducing the translational fidelity of the ribosome; reducing the efficiency of the translational termination machinery; inhibiting NMD; interfering with the regulation of gene transcription or translation or stabilising mRNA against NMD (Kellermayer, R European Journal of Medical Genetics 2006, 49, 445-450, Belgrader, P. J. C., and Maquat, L.E., Proc. Natl. Acad. Sci. USA 1993, 90, 482-486).

[0010] Agents which allow translational read-through of PTC mutations have the potential to treat any disease where the presence of a PTC in a gene is directly causative, a risk factor or an aggravating factor in disease.

[0011] Aminoglycosides (such as paromomycin, gentamicin, G418 / geneticin) are a class of antibiotics able to reduce translational fidelity. They exert their antibacterial action through inhibition of prokaryotic protein synthesis, however, at sub-lethal doses they lower translational fidelity of ribosomes leading to an increased rate of mis-incorporation of amino acids and also to an increased rate of translational read-through of stop codons (Davies, J. & Gorini, L., Proc. Natl. Acad. Sci. USA 1964, 51 , 883-8, Weinstein, Proc. Natl. Acad. Sci. USA 1964, 52, 988-996).

[0012] Aminoglycosides have been shown to allow read-through of PTC mutations in the CFTR gene responsible for cystic fibrosis in vitro (Bedwell, D.M., et al., Nature Medicine 1997, 3, 1280-1284) and in vivo, including patients (Ming Du, et al., Journal of Molecular Medicine 2002, 80, 595-604, Clancy, J.P. et al., Am J Respir Crit Care Med 2001 , 163, 1683- 1692, Wilschanski, M., et al., The New England Journal of Medicine 2003, 139, 1433- 1441).

[0013] In an animal model of Duchenne muscular dystrophy (DMD), 7 of 8 male mdx mice treated with 34 mg / kg gentamicin showed increased protection from contractile injury and 10-20% restoration of wild-type dystrophin levels (Barton-Davis, E.R., et al., Journal of Clinical Investigation 1999, 104, 375-381).

[0014] Gentamicin treatment induced functional type VII collagen in recessive dystrophic epidermolysis bullosa patients (Woodley, D. T. et al. Journal of Clinical Investigation 2017, 127, 3028-3038) with in vitro evidence supporting a read-through mechanism (Has, C. et al. Journal of Investigative Dermatology 2022, 142, 1227-1230).

[0015] The read-through of genes containing PTC mutations relevant to cystinosis, Hurler syndrome, inherited blindness and cancer genes have all been demonstrated following aminoglycoside treatment (Helip- Wooley, A., et al., Molecular Genetics and Metabolism 2002, 75, 128-133, Keeling, K.M., et al., Human Molecular Genetics 2001 , 10, 291-299, Moosajee, M.K., et al., Human Molecular Genetics 2008,17, 3987-4000). However, aminoglycosides are not suitable as PTC read-through agents due to the severe toxicity observed with high doses or repeated treatments.

[0016] WO2021228945A1 describes compounds, including TLN486 (TLN68), for use in preventing and / or treating a disease caused by a nonsense mutation.1-(4-(furan-3-yl)-6,7-dihydro-3- / - 1-(3-(dimethylamino)propyl)-2- 1-(3-(dimethylamino)propyl)-1- cyclopenta[d]pyrimidin-2-yl)-3-(3- methyl-3-(4-methylquinazolin-2- methyl-3-(4-methylquinazolin-2-(pi peridin-1 -yl)propyl)guanidine yl)guanidine yl)guanidine

[0017] Christie, M., et al., Bioorganic & Medicinal Chemistry Letters 2022 describes compounds, including 1-(4-(furan-3-yl)-6,7-dihydro-5 / 7-cyclopenta[d]pyrimidin-2-yl)-3-(3- (piperidin-1-yl)propyl)guanidine that promote readthrough of premature termination codons in cells. Interestingly, they report that alkylated guanidines, including for example 1-(3- (dimethylamino)propyl)-2-methyl-3-(4-methylquinazolin-2-yl)guanidine and 1 -(3- (dimethylamino)propyl)-1-methyl-3-(4-methylquinazolin-2-yl)guanidine, are inactive as readthrough agents.

[0018] Certain compounds of the present invention are suitable for use in treating diseases / conditions which are associated with PTC mutations. Certain compounds of the invention show more potent read-through activity when compared to prior art compounds. Certain compounds of the invention show better permeability when compared to prior art compounds. Certain compounds of the invention are more orally bioavailable when compared to prior art compounds.

[0019] A structurally novel class of compounds has now been found which provides PTC read-through agents.BRIEF SUMMARY OF THE DISCLOSURE

[0020] In accordance with a first aspect, the present invention provides a compound of formula (I), or a pharmaceutically acceptable salt or N-oxide thereof:whereinL1is independently Ci-Ce-alkylene;R1is independently selected from phenyl, naphthyl, and 5-, 6-, 8-, 9- or 10- membered heteroaryl; wherein R1is optionally substituted with from 1 to 5 R5groups;R2is independently selected from H, halo, nitro, cyano, NR7R9, OR7, SR7, SOR7, S(O)2R7, SO2NR7R7, CO2R7, C(O)R7, CONR7R7, Ci-C4-alkyl, C2-C4-alkenyl, C2-C4-alkynyl, Ci-C4- haloalkyl and cyclopropyl;R3and R4are each independently selected from H, Ci-Ce-alkyl, Ci-Ce-haloalkyl, Cs-Cs cycloalkyl, 3- to 8-membered heterocycloalkyl, Ci-C4-alkylene-R3a, C2-C4-alkylene-R3b; or wherein R3and R4, together with the nitrogen atom to which they are attached, form a 5- to 8-membered heterocycloalkyl ring, optionally substituted with 1 to 4 R6groups;R3ais independently selected from C(O)NR7R7, SC>2NR7R7, CO2R7, Cs-Cs cycloalkyl, Cs-Cs- cycloalkenyl, phenyl, 3- to 8-membered heterocycloalkyl, 5- to 8-membered heterocycloalkenyl and 5- or 6-membered heteroaryl; wherein where R3ais cycloalkyl, cycloalkenyl, heterocycloalkyl or heterocycloalkenyl, R3ais optionally substituted with from 1 to 4 R6groups and where R3ais phenyl or heteroaryl, R3ais optionally substituted with from 1 to 5 R8groups;R3bis independently selected from NR7R9and OR7;R5is independently at each occurrence selected from halo, nitro, cyano, NR7R9, OR7, SR7, SOR7, S(O)2R7, S(O)2NR7R7, CO2R7, C(O)R7, C(O)NR7R7, Ci-C4-alkyl, C2-C4-alkenyl, C2-C4- alkynyl, Ci-C4-haloalkyl, phenyl, 5- or 6- membered heteroaryl, Cs-Cs cycloalkyl, Cs-Cs- cycloalkenyl, 3- to 8-membered heterocycloalkyl and 5- to 8-membered heterocycloalkenyl; wherein where R5is cycloalkyl, cycloalkenyl, heterocycloalkyl or heterocycloalkenyl, R5is optionally substituted with from 1 to 4 R6groups and where R5is phenyl or heteroaryl, R5is optionally substituted with from 1 to 5 R8groups;R6is independently at each occurrence selected from =0, =S, halo, nitro, cyano, NR7aR9, OR7, SR7, SOR7, S(O)2R7, SO2NR7aR7a, CO2R7, C(O)R7, CONR7aR7a, Ci-C4-alkyl, C2-C4- alkenyl, C2-C4-alkynyl, Ci-C4-haloalkyl and cyclopropyl;R7and R7aare each independently at each occurrence selected from H and C1-C4 alkyl; or two R7groups, together with the nitrogen atom to which they are attached, form a 5- to 8- membered heterocycloalkyl ring, optionally substituted with 1 to 4 R6groups;R8is independently at each occurrence selected from halo, nitro, cyano, NR7R9, OR7, SR7, SOR7, S(O)2R7, SO2NR7R7, CO2R7, C(O)R7, CONR7R7, Ci-C4-alkyl, C2-C4-alkenyl, C2-C4- alkynyl, Ci-C4-haloalkyl and cyclopropyl;R9is independently at each occurrence selected from H, C1-C4 alkyl and C(O)-Ci-C4-alkyl; or R7and R9, together with the nitrogen atom to which they are attached, form a 5- to 8- membered heterocycloalkyl ring, optionally substituted with 1 to 4 R6groups; wherein any of the aforementioned alkyl, alkylene, alkenyl, or cyclopropyl groups is optionally substituted, where chemically possible, by 1 to 5 substituents which are each independently at each occurrence selected from the group consisting of: Ci-C4-alkyl, oxo, halo, nitro, cyano,NRaRb, ORa, SRa, CO2Ra, C(O)Ra, CONRaRa, S(O)Ra, and S(O)2Ra; wherein Rais independently at each occurrence selected from H, and Ci-C4-alkyl; and Rbis independently at each occurrence selected from H, Ci-C4-alkyl, C(O)-Ci-C4-alkyl and S(O)2-Ci-C4-alkyl.

[0021] In an embodiment, the compound of formula (I) is a compound of formula (II):wherein L1, R2, R3, R4and R5are as defined for formula (I); and wherein R5ais independently selected from H, halo, nitro, cyano, NR7R9, OR7, SR7, SOR7, S(O)2R7, S(O)2NR7R7, CO2R7, C(O)R7, C(O)NR7R7, Ci-C4-alkyl, C2-C4-alkenyl, C2-C4-alkynyl, Ci-C4-haloalkyl; and n is an integer from 0 to 4. R5amay be independently selected from halo, nitro, cyano, NR7R9, OR7, SR7, SOR7, S(O)2R7, S(O)2NR7R7, CO2R7, C(O)R7, C(O)NR7R7, Ci-C4-alkyl, C2-C4-alkenyl, C2-C4-alkynyl, Ci-C4-haloalkyl; and n is an integer from 0 to 4.

[0022] In an embodiment, the compound of formula (I) is a compound of formula (III):wherein L1, R2, R3, R4and R5are as defined for formula (I); and wherein X1is independently selected from N and CR5b; X2is independently selected from N and CR5c; R5band R5care each independently selected from H, halo, nitro, cyano, NR7R9, OR7, SR7, SOR7, S(O)2R7, S(O)2NR7R7, CO2R7, C(O)R7, C(O)NR7R7, Ci-C4-alkyl, C2-C4-alkenyl, C2-C4-alkynyl, C1-C4- haloalkyl; and m is an integer from 0 to 3.

[0023] In an embodiment, the compound of formula (I) is a compound of formula (IV):wherein L1, R2, R3, R4and R5are as defined for formula (I); and wherein X2is independently selected from N and CR5c; R5cis independently selected from H, halo, nitro, cyano, NR7R9, OR7, SR7, SOR7, S(O)2R7, S(O)2NR7R7, CO2R7, C(O)R7, C(O)NR7R7, Ci-C4-alkyl, C2-C4- alkenyl, C2-C4-alkynyl, Ci-C4-haloalkyl; and m is an integer from 0 to 3. In these embodiments, X2may be N.

[0024] In an embodiment, the compound of formula (I) is a compound of formula (V):wherein L1, R2, R3, R4and R5are as defined for formula (I); and wherein X1is independently selected from N and CR5b; R5band R5care each independently selected from H, halo, nitro, cyano, NR7R9, OR7, SR7, SOR7, S(O)2R7, S(O)2NR7R7, CO2R7, C(O)R7, C(O)NR7R7, Ci-C4- alkyl, C2-C4-alkenyl, C2-C4-alkynyl, Ci-C4-haloalkyl; and m is an integer from 0 to 3. In these embodiments, X1may be N. In alternative embodiments, X1may be CR5b, e.g. CH.

[0025] In an embodiment, the compound of formula (I) is a compound of formula (VI):wherein L1, R1, R3and R4are as defined for formula (I).

[0026] In an embodiment, the compound of formula (I) is a compound of formula (VII):wherein R1, R2, R3and R4are as defined for formula (I).

[0027] In an embodiment, the compound of formula (I) is a compound of formula (VIII):wherein R1, R3and R4are as defined for formula (I).

[0028] In an embodiment, the compound of formula (I) is a compound of formula (IX):wherein R3, R4and R5are as defined for formula (I); and wherein R5ais independently selected from H, halo, nitro, cyano, NR7R9, OR7, SR7, SOR7, S(O)2R7, S(O)2NR7R7, CO2R7, C(O)R7, C(O)NR7R7, Ci-C4-alkyl, C2-C4-alkenyl, C2-C4-alkynyl, Ci-C4-haloalkyl; and n is an integer from 0 to 4. R5amay be independently selected from halo, nitro, cyano, NR7R9, OR7, SR7, SOR7, S(O)2R7, S(O)2NR7R7, CO2R7, C(O)R7, C(O)NR7R7, Ci-C4-alkyl, C2-C4-alkenyl, C2-C4-alkynyl, Ci-C4-haloalkyl; and n is an integer from 0 to 4.

[0029] In an embodiment, the compound of formula (I) is a compound of formula (X):wherein R3, R4and R5are as defined for formula (I); and wherein X1is independently selected from N and CR5b; X2is independently selected from N and CR5c; R5band R5care each independently selected from H, halo, nitro, cyano, NR7R9, OR7, SR7, SOR7, S(O)2R7, S(O)2NR7R7, CO2R7, C(O)R7, C(O)NR7R7, Ci-C4-alkyl, C2-C4-alkenyl, C2-C4-alkynyl, C1-C4- haloalkyl; and m is an integer from 0 to 3.

[0030] In an embodiment, the compound of formula (I) is a compound of formula (XI):wherein R3, R4and R5are as defined for formula (I); and wherein X2is independently selected from N and CR5c; R5cis independently selected from H, halo, nitro, cyano, NR7R9, OR7, SR7, SOR7, S(O)2R7, S(O)2NR7R7, CO2R7, C(O)R7, C(O)NR7R7, Ci-C4-alkyl, C2-C4-alkenyl, C2-C4- alkynyl, Ci-C4-haloalkyl; and m is an integer from 0 to 3. In these embodiments, X2may be N.

[0031] In an embodiment, the compound of formula (I) is a compound of formula (XII):wherein R3, R4and R5are as defined for formula (I); and wherein X1is independently selected from N and CR5b; R5band R5care each independently selected from H, halo, nitro, cyano, NR7R9, OR7, SR7, SOR7, S(O)2R7, S(O)2NR7R7, CO2R7, C(O)R7, C(O)NR7R7, Ci-C4-alkyl, C2- C4-alkenyl, C2-C4-alkynyl, Ci-C4-haloalkyl; and m is an integer from 0 to 3. In these embodiments, X1may be N. In alternative embodiments, X1may be CR5b, e.g. CH.

[0032] In an embodiment, the compound of formula (I) is a compound of formula (XIII):wherein L1, R2, R3, R4and R5are as defined for formula (I); and wherein R5ais independently selected from H, halo, nitro, cyano, NR7R9, OR7, SR7, SOR7, S(O)2R7, S(O)2NR7R7, CO2R7, C(O)R7, C(O)NR7R7, Ci-C4-alkyl, C2-C4-alkenyl, C2-C4-alkynyl, Ci-C4-haloalkyl; and p is an integer from 0 to 6. R5amay be independently selected from halo, nitro, cyano, NR7R9, OR7,SR7, SOR7, S(O)2R7, S(O)2NR7R7, CO2R7, C(O)R7, C(O)NR7R7, Ci-C4-alkyl, C2-C4-alkenyl, C2-C4-alkynyl, Ci-C4-haloalkyl; and n is an integer from 0 to 4.

[0033] In an embodiment, the compound of formula (I) is a compound of formula (XIV):wherein R2, R3, R4and R5are as defined for formula (I); and wherein R5ais independently selected from H, halo, nitro, cyano, NR7R9, OR7, SR7, SOR7, S(O)2R7, S(O)2NR7R7, CO2R7, C(O)R7, C(O)NR7R7, Ci-C4-alkyl, C2-C4-alkenyl, C2-C4-alkynyl, Ci-C4-haloalkyl; and p is an integer from 0 to 6.

[0034] The following embodiments apply to compounds of any of formulae (l)-(XIV). These embodiments are independent and interchangeable. Any one embodiment may be combined with any other embodiment, where chemically allowed. In other words, any of the features described in the following embodiments may (where chemically allowable) may be combined with the features described in one or more other embodiments. In particular, where a compound is exemplified or illustrated in this specification, any two or more of the embodiments listed below, expressed at any level of generality, which encompass that compound may be combined to provide a further embodiment which forms part of the present disclosure.

[0035] R1may be independently selected from 5-, 6-, 8-, 9- or 10- membered heteroaryl. R1may be a heteroaryl group comprising from 1 to 2 nitrogen atoms in the ring system. R1may be a 6- or 10- membered heteroaryl group comprising from 1 to 2 nitrogen atoms in the ring system.

[0036] R1may be independently selected from phenyl and naphthyl.

[0037] R1may be a bicyclic group. R1may be naphthyl. R1may be 8-, 9- or 10- membered heteroaryl, e.g. 8-, 9- or 10- membered heteroaryl comprising from 1 to 2 nitrogen atoms in the ring system.

[0038] R1may be a monocyclic group. R1may be phenyl. R1may be 5- or 6-membered heteroaryl, e.g. 5- or 6-membered heteroaryl comprising from 1 to 2 nitrogen atoms in the ring system.

[0039] R1may be a 10-membered heteroaryl, e.g. a 10-membered heteroaryl comprising from 1 to 2 nitrogen atoms in the ring system. R1may be quinazoline. R1may have the structure:wherein R5ais independently selected from H, halo, nitro, cyano, NR7R9, OR7, SR7, SOR7, S(O)2R7, S(O)2NR7R7, CO2R7, C(O)R7, C(O)NR7R7, C1-C4- alkyl, C2-C4-alkenyl, C2-C4-alkynyl, Ci-C4-haloalkyl; and n is an integer from 0 to 4. R5amay be independently selected from halo, nitro, cyano, NR7R9, OR7, SR7, SOR7, S(O)2R7, S(O)2NR7R7, CO2R7, C(O)R7, C(O)NR7R7, Ci-C4-alkyl, C2-C4-alkenyl, C2-C4-alkynyl, C1-C4- haloalkyl; and n is an integer from 0 to 4.

[0040] R5amay be independently selected from H, cyano, nitro, OR7, NR7R9, Ci-C4-alkyl, Ci-C4-haloalkyl, and cyclopropyl. R5amay be independently selected from H, cyano, nitro, NR7R9, Ci-C4-alkyl, Ci-C4-haloalkyl, and cyclopropyl. R5amay be independently selected from H, Ci-C4-alkyl, Ci-C4-haloalkyl, and cyclopropyl. R5amay be independently selected from H, and Ci-C4-alkyl. R5amay be independently selected from cyano, nitro, OR7, NR7R9, Ci-C4-alkyl, Ci-C4-haloalkyl, and cyclopropyl. R5amay be independently selected from cyano, nitro, NR7R9, Ci-C4-alkyl, Ci-C4-haloalkyl, and cyclopropyl. R5amay be independently selected from Ci-C4-alkyl, Ci-C4-haloalkyl, and cyclopropyl. R5amay be Ci-C4-alkyl. R5amay be independently selected from Ci-C4-alkyl and OR7. R5amay be Ci-C2-alkyl, e.g. methyl or ethyl. R5amay be Ci-alkyl, e.g. methyl. R5amay be H.

[0041] n may be 0. n may be a integer from 1 to 4. n may be 1.

[0042] R1may be 6-membered group. R1may be pyridine, e.g. pyridin-2-yl. R1may be pyrimidine, e.g. pyrimidin-2-yl. R1may have the structure:wherein X1is independently selected from N and CR5b; X2is independently selected from N and CR5c; R5band R5care each independently selected from H, halo, nitro, cyano, NR7R9, OR7, SR7, SOR7, S(O)2R7, S(O)2NR7R7, CO2R7, C(O)R7, C(O)NR7R7, Ci-C4-alkyl, C2-C4-alkenyl, C2-C4-alkynyl, Ci-C4-haloalkyl; and m is an integer from 0 to 3. X1may be N. R1may have the structure:[ X1and X2may each be N. R1may have the structure:

[0044] X2may be CR5c. R1may have the structure:[ R1may have the structure:

[0046] X1may be CR5b, e.g. CH. R1may have the structure:, . .R5bmay be independently selected from H, halo, Ci-C4-alkyl, Ci-C4-haloalkyl, and cyclopropyl. R5bmay be independently selected from H, halo and Ci-C4-alkyl. R5bmay be independently selected from halo, Ci-C4-alkyl and OR7. R5bmay be Ci-C2-alkyl, e.g. methyl or ethyl. R5bmay be Ci-alkyl, e.g. methyl. R5bmay be H.R5cmay be independently at each occurrence selected from halo, nitro, cyano, NR7R9, OR7, SR7, SOR7, S(O)2R7, S(O)2NR7R7, CO2R7, C(O)R7, C(O)NR7R7, Ci-C4-alkyl, C2-C4-alkenyl, C2-C4-alkynyl. Thus, it may be that R5cis not H. Alternatively, R5cmay be H. It may be that R5cis ON.

[0047] m may be 0. m may be an integer from 1 to 3. m may be 1 or 2. m may be 1.

[0048] R1may be a 9-membered group. R1may have the structure:wherein X1is independently selected from N and CR5b; X2is independently selected from N and CR5c; R5band R5care each independently selected from H, halo, nitro, cyano, NR7R9, OR7, SR7, SOR7, S(O)2R7, S(O)2NR7R7, CO2R7, C(O)R7, C(O)NR7R7, Ci-C4-alkyl, C2-C4- alkenyl, C2-C4-alkynyl, Ci-C4-haloalkyl; and p is an integer from 0 to 6. X1may be N. R1may have the structure:

[0050] p may be 0. p may be an integer from 1 to 6. p may be an integer from 1 to 4. p may be an integer from 1 to 3. p may be 1. p may be 2. p may be 3.

[0051] It may be that R5is independently at each occurrence selected from halo, nitro, cyano, NR7R9, OR7, SR7, SOR7, S(O)2R7, S(O)2NR7R7, CO2R7, C(O)R7, C(O)NR7R7, C1-C4- alkyl, C2-C4-alkenyl, C2-C4-alkynyl, Ci-C4-haloalkyl. R5may be independently at each occurrence selected from halo, cyano, nitro, OR7, NR7R9, Ci-C4-alkyl, Ci-C4-haloalkyl, andcyclopropyl. R5may be independently at each occurrence selected from halo, OR7, C1-C4- alkyl and Ci-C4-haloalkyl.

[0052] Where m is 1 or more, it may be that in a single occurrence R5is selected from phenyl, 5- or 6- membered heteroaryl, C3-C8 cycloalkyl, Cs-Cs-cycloalkenyl, 3- to 8- membered heterocycloalkyl and 5- to 8-membered heterocycloalkenyl; wherein where R5is cycloalkyl, cycloalkenyl, heterocycloalkyl or heterocycloalkenyl, R5is optionally substituted with from 1 to 4 R6groups and where R5is phenyl or heteroaryl, R5is optionally substituted with from 1 to 5 R8groups. Where m is 2 or more, it may be that in a single occurrence R5is selected from phenyl, 5- or 6- membered heteroaryl, C3-C8 cycloalkyl, Cs-Cs-cycloalkenyl, 3- to 8-membered heterocycloalkyl and 5- to 8-membered heterocycloalkenyl; wherein where R5is cycloalkyl, cycloalkenyl, heterocycloalkyl or heterocycloalkenyl, R5is optionally substituted with from 1 to 4 R6groups and where R5is phenyl or heteroaryl, R5is optionally substituted with from 1 to 5 R8groups; and in other occurrences, R5is selected from halo, nitro, cyano, NR7R9, OR7, SR7, SOR7, S(O)2R7, S(O)2NR7R7, CO2R7, C(O)R7, C(O)NR7R7, Ci-C4-alkyl, C2-C4-alkenyl, C2-C4-alkynyl, Ci-C4-haloalkyl.

[0053] It may be that R2is independently selected from H, halo, Ci-C4-alkyl and C1-C4- haloalkyl. R2may be H. R2may be independently selected from halo, Ci-C4-alkyl and C1- C4-haloalkyl. R2may be Ci-C4-alkyl. R2may be Ci-C4-haloalkyl. R2may be halo.

[0054] L1may be -Ci-C4-alkylene-. L1may be -C2-C4-alkylene-. L1may be -C2-alkylene. L1may be unsubstituted. L1may be -CH2CH2-. L1may be substituted with an oxo group on the carbon to which the NR3R4group is attached. L1may be -CH2CH2C(O)-.

[0055] R4may be independently selected from H, Ci-Ce-alkyl and Ci-Ce-haloalkyl. R4may be independently selected from H and Ci-C4-alkyl. R4may be H. R4may be Ci-C4-alkyl.

[0056] R3may be independently selected from Ci-Ce-alkyl, Ci-Ce-haloalkyl, C3-C8 cycloalkyl, 3- to 8-membered heterocycloalkyl, Ci-C4-alkylene-R3a, C2-C4-alkylene-R3b. R3may be selected from Ci-Ce-alkyl and Ci-Ce-haloalkyl. R3may be selected from Ci-C4-alkyl and Ci-C4-haloalkyl. R3may be Ci-C4-alkyl, e.g. methyl, ethyl or isopropyl. R3may be C1- C4-haloalkyl, e.g. 2,2,2-trifluoroethyl. R3may be C2-C4-alkylene-R3b. R3bmay be OR7, e.g. O-Ci-C4-alkyl, e.g. OMe. R3may be selected from C3-C8 cycloalkyl, 3- to 8-membered heterocycloalkyl and Ci-C4-alkylene-R3a; wherein R3ais selected from Cs-Cs-cycloalkyl and 3- to 8-membered heterocycloalkyl.

[0057] It may be that R3and R4are each Ci-C4-alkyl. It may be that R3and R4are each methyl.

[0058] It may be that R3and R4, together with the nitrogen atom to which they are attached, form a 5- to 8-membered heterocycloalkyl ring, optionally substituted with 1 to 4 R6groups. It may be that R3and R4, together with the nitrogen atom to which they are attached, form a 6-membered heterocycloalkyl ring, optionally substituted with 1 to 4 R6groups. It may be that R3and R4, together with the nitrogen atom to which they are attached, form a morpholine ring, optionally substituted with 1 to 4 R6groups.

[0059] R6may be independently at each occurrence selected from =0, NR7aR9, OR7, CO2R7, C(O)R7, CONR7aR7a, Ci-C4-alkyl.

[0060] R7may be independently at each occurrence H. R7may be independently at each occurrence C1-C4 alkyl. R7may be independently at each occurrence C1-C2 alkyl.

[0061] R7amay be independently at each occurrence H. R7amay be independently at each occurrence C1-C4 alkyl. R7amay be independently at each occurrence C1-C2 alkyl.

[0062] R8may be independently at each occurrence selected from halo, nitro, cyano, NR7R9, OR7, SR7, SOR7, S(O)2R7, SO2NR7R7, CO2R7, C(O)R7, CONR7R7, Ci-C4-alkyl, C1-C4- haloalkyl and cyclopropyl. R8may be independently at each occurrence selected from halo, nitro, cyano, NR7R9, OR7, Ci-C4-alkyl, Ci-C4-haloalkyl and cyclopropyl. R8may be independently at each occurrence selected from halo, OR7, Ci-C4-alkyl and Ci-C4-haloalkyl.

[0063] R9may be independently H. R9may be independently C1-C4 alkyl. R9may be independently C1-C2 alkyl. R9may be independently C(O)-Ci-C4-alkyl. R9may be independently C(O)-Ci-C2-alkyl.

[0065] In an embodiment, the compound of formula (I) is not a compound selected fromList A:

[0066] In accordance with a second aspect, the present invention provides a pharmaceutical composition comprising a compound defined in the first aspect, and one or more pharmaceutically acceptable excipients. In embodiments of the second aspect, the compounds of List A are excluded. In embodiments of the second aspect, the compounds of List A are not excluded.

[0067] In accordance with a third aspect, the present invention provides a compound as defined in the first aspect or a pharmaceutical composition as defined in the second aspect, for use as a medicament. In embodiments of the third aspect, the compounds of List A are excluded. In embodiments of the third aspect, the compounds of List A are not excluded.

[0068] In accordance with a fourth aspect, the present invention provides the use of a compound as defined in the first aspect or a pharmaceutical composition as defined in the second aspect, for the manufacture of a medicament. In embodiments of the fourth aspect, the compounds of List A are excluded. In embodiments of the fourth aspect, the compounds of List A are not excluded.

[0069] In accordance with a fifth aspect, the present invention provides a compound as defined in the first aspect, or a pharmaceutical composition as defined in the second aspect, for use in a method of treatment or prophylaxis of a disease of the musculoskeletal system, a disease of the skin, a metabolic disease, a disease of the Nervous System, a Cardiovascular disease, an Endocrine disorder, a disease of the eye, a disease affecting the urogenital system, a haemic or lymphatic condition, a respiratory disease, an inflammatory or autoimmune condition, a disease of the Gastrointestinal system, a Neoplasm, cancer, or a disease or disorder selected from Amelogenesis Imperfecta, Anodontia, Odontodysplasia, Branchio-Oto-Renal Syndrome, Sotos Syndrome, and Waardenburg's Syndrome. In embodiments of the fifth aspect, the compounds of List A are excluded. In embodiments of the fifth aspect, the compounds of List A are not excluded.

[0070] In accordance with a sixth aspect, the present invention provides a method for the treatment or prophylaxis of a disease of the musculoskeletal system, a disease of the skin, a metabolic disease, a disease of the Nervous System, a Cardiovascular disease, an Endocrine disorder, a disease of the eye, a disease affecting the urogenital system, a haemic or lymphatic condition, a respiratory disease, an inflammatory or autoimmune condition, a disease of the Gastrointestinal system, a Neoplasm, cancer, or a disease or disorder selected from Amelogenesis Imperfecta, Anodontia, Odontodysplasia, Branchio-Oto-Renal Syndrome, Sotos Syndrome, and Waardenburg's Syndrome, said method comprising administering to a subject, an effective amount of a compound as defined in the first aspect, or a pharmaceutical composition as defined in the second aspect. In embodiments of the sixth aspect, the compounds of List A are excluded. In embodiments of the sixth aspect, the compounds of List A are not excluded.

[0071] In accordance with a seventh aspect, the present invention provides a compound as defined in the first aspect, or a pharmaceutical composition as defined in the second aspect, for use in a method of treatment or prophylaxis of a disease selected from: Recessivedystrophic epidermolysis bullosa, Junctional epidermolysis bullosa Xeroderma pigmentosum, Netherton syndrome, Duchenne Muscular Dystrophy, Becker Muscular Dystrophy, Cystic Fibrosis, Alport Syndrome, Dravet Syndrome, Aniridia, Methylmalonic Acidemia, Colorectal Cancer, Endometrium Cancer, Breast Cancer, Ovarian Cancer, Lung Squamous Cell Carcinoma, Head and Neck Squamous Cell Carcinoma, Familial adenomatous polyposis, Hemophilia A, Hemophilia B, Choroideremia, Pulmonary Artery Hypertension, Ataxia telangiectasia, Shwachman-Diamond syndrome, Mucopolysaccharidosis Type I, Mucopolysaccharidosis Type VI, Mucopolysaccharidosis type III, Niemann-Pick Disease, Primary Ciliary Dyskinesia, Usher syndrome, and Retinitis Pigmentosa. In embodiments of the seventh aspect, the compounds of List A are excluded. In embodiments of the seventh aspect, the compounds of List A are not excluded.

[0072] In accordance with an eighth aspect, the present invention provides a method for the treatment or prophylaxis of a disease selected from: Recessive dystrophic epidermolysis bullosa, Junctional epidermolysis bullosa Xeroderma pigmentosum, Netherton syndrome, Duchenne Muscular Dystrophy, Becker Muscular Dystrophy, Cystic Fibrosis, Alport Syndrome, Dravet Syndrome, Aniridia, Methylmalonic Acidemia, Colorectal Cancer, Endometrium Cancer, Breast Cancer, Ovarian Cancer, Lung Squamous Cell Carcinoma, Head and Neck Squamous Cell Carcinoma, Familial adenomatous polyposis, Hemophilia A, Hemophilia B, Choroideremia, Pulmonary Artery Hypertension, Ataxia telangiectasia, Shwachman-Diamond syndrome, Mucopolysaccharidosis Type I, Mucopolysaccharidosis Type VI, Mucopolysaccharidosis type III, Niemann-Pick Disease, Primary Ciliary Dyskinesia, Usher syndrome, and Retinitis Pigmentosa. In embodiments of the eighth aspect, the compounds of List A are excluded. In embodiments of the eighth aspect, the compounds of List A are not excluded.

[0073] In accordance with a ninth aspect, the present invention provides the use of a compound as defined in the first aspect, or a pharmaceutical composition as defined in the second aspect for the manufacture of a medicament for the treatment or prophylaxis of a disease of the musculoskeletal system, a disease of the skin, a metabolic disease, a disease of the Nervous System, a Cardiovascular disease, an Endocrine disorder, a disease of the eye, a disease affecting the urogenital system, a haemic or lymphatic condition, a respiratory disease, an inflammatory or autoimmune condition, a disease of the Gastrointestinal system, a Neoplasm, cancer, or a disease or disorder selected from Amelogenesis Imperfecta, Anodontia, Odontodysplasia, Branchio-Oto-Renal Syndrome, Sotos Syndrome, and Waardenburg's Syndrome. In embodiments of the ninth aspect, the compounds of List A are excluded. In embodiments of the ninth aspect, the compounds of List A are not excluded.

[0074] In accordance with a tenth aspect, the present invention provides a method of allowing translational read-through of PTC mutations in a subject, said method comprising administering to a subject an effective amount of a compound as defined in the first aspect, or a pharmaceutical composition as defined in the second aspect. In embodiments of the tenth aspect, the compounds of List A are excluded. In embodiments of the tenth aspect, the compounds of List A are not excluded.

[0075] In accordance with an eleventh aspect, the present invention provides a compound as defined in the first aspect, or a pharmaceutical composition as defined in the second aspect, for use in a method of treating conditions or disorders which are associated with PTC mutations in a subject, said method comprising administering to a subject an effective amount of a compound as defined in the first aspect, or a pharmaceutical composition as defined in the second aspect. In embodiments of the eleventh aspect, the compounds of List A are excluded. In embodiments of the eleventh aspect, the compounds of List A are not excluded.

[0076] In accordance with a twelfth aspect, the present invention provides a method of treating conditions or disorders which are associated with PTC mutations in a subject, said method comprising administering to a subject an effective amount of a compound as defined in the first aspect, or a pharmaceutical composition as defined in the second aspect. In embodiments of the twelfth aspect, the compounds of List A are excluded. In embodiments of the twelfth aspect, the compounds of List A are not excluded.

[0077] Suitably a PTC mutation may be any mutation which generates a premature termination codon (PTC) in a gene. Suitably a PTC mutation may be any mutation which generates an in-frame premature termination codon (PTC) in a gene. Suitably a PTC mutation occurs in the nucleotide sequence of a gene. Suitably a PTC mutation may occur in the coding region or non-coding region of a gene. Suitably a PTC mutation may occur in the coding region of a gene.

[0078] Suitably the mutation may be a point mutation, for example a substitution, deletion or insertion mutation. Suitably the mutation may be a substitution mutation. Suitably the substitution mutation may replace one nucleotide with another different nucleotide, suitably within the nucleotide sequence of a gene. Suitably the mutation may therefore be regarded as a nonsense mutation. Suitably the mutation may be a nonsense mutation which causes an in-frame PTC in a gene. Suitably the PTC may be any known termination codon such as TAG, TAA or TGA. Suitably the PTC may be TGA. Suitably therefore in some embodiments, the PTC mutation may be a substitution mutation which generates a PTC comprising TAG,TAA or TGA in a gene. Suitably therefore in some embodiments, the PTC mutation may be a substitution mutation of CGA to TGA or CAG to TAG in a gene.

[0079] A condition or disorder associated with a PTC mutation in a subject may be a condition or disorder which is directly or indirectly caused by a PTC mutation in a subject. Suitably a condition or disorder associated with a PTC mutation in a subject may be a condition or disorder which is directly caused by a PTC mutation in a subject. Suitably a condition or disorder associated with a PTC mutation in a subject may be a condition or disorder which is caused by a PTC mutation in one or more genes in a subject. Suitably a condition or disorder associated with a PTC mutation in a subject may be a condition or disorder which is directly caused by a PTC mutation in one or more genes in a subject. Suitably a condition or disorder associated with a PTC mutation in a subject may be a condition or disorder which is caused by a PTC mutation in one or more genes in a subject leading to a loss of function of the or each gene. Suitably reference herein to ‘a’ PTC mutation may be understood to refer to one or more PTC mutations, or a plurality of PTC mutations, suitably which may be present in one or more genes in a subject.

[0080] Suitably the one or more genes in which the PTC mutation is present are genes relating to one or more diseases as described herein. Suitably the one or more genes may be directly or indirectly linked with one or more diseases described herein. Suitably the one or more genes may contribute to, or cause, the phenotype of one or more diseases described herein. Such genes may be for example LAMB3, COL.7A, COL4, CFTR, and DMD, which are each associated with one or more of the diseases identified herein. In some embodiments, for example in which the disease is cancer, the one or more genes in which the PTC mutation is present may be cell cycle genes, tumour suppressor genes etc. Such tumour suppressor genes may include, for example, TP53, PTEN, APC, ARID1 A or CTCF. Suitably such mutations may be regarded as driver mutations, suitably which drive a disease phenotype, such as tumour growth. Therefore, in some embodiments, the PTC mutation may be regarded as a driver mutation. In other embodiments, the one or more genes in which the PTC mutation is present may not contribute to or cause the disease phenotype. Suitably such mutations may be regarded as bystander mutations. Therefore in some embodiments, the PTC mutation may be regarded as a bystander mutation.

[0081] Conditions or disorders associated with a PTC mutation are well known in the art, and suitable such disorders which may be treated or prevented by the present invention are described hereinbelow. However, suitably, such disorders that are associated with PTC mutations may also be identified or determined by the use of standard molecular biology techniques. Suitably a condition or disorder may be determined as being associated with aPTC mutation by the use of molecular biology techniques, such as sequencing, single strand conformational polymorphism, denaturing gradient gel electrophoresis, heteroduplex analysis, or restriction fragment length polymorphism. Suitably a PTC mutation present in one or more genes of a subject which may cause a disease or condition can be identified using such techniques. Suitably, whole genome sequencing or whole exome sequencing may be used to identify a PTC mutation in one or more genes of a subject, suitably which may cause a disease or condition. Suitable methodologies may be described in Karagiannakos et al. Cancers (Basel) 2022, 14, 664, Stark et al. ‘A prospective evaluation of whole-exome sequencing as a first-tier molecular test in infants with suspected monogenic disorders’ Genetics in Medicine, Volume 18, Issue 11 , 2016, North et al. ‘Approach to the diagnosis of congenital myopathies’ Neuromuscular Disorders, Volume 24, Issue 2, 2014, for example.

[0082] Suitably such techniques may be applied to a sample obtained from a subject suffering from a disease or condition or suspected of suffering from a disease or condition, suitably which may be caused by a PTC mutation. Suitably such techniques may comprise comparing a result from said sample, to a result obtained from a reference sample. Suitably a reference sample may be a sample of the same type from a healthy subject, suitably from an equivalent healthy subject of the same age, nationality, race, height, weight etc. Suitable samples may include: a blood sample, serum sample, CNS fluid sample, tissue sample, cell sample, and the like. Suitably comparing a result in the context of the techniques above may comprise comparing the sequence of one or more genes from the subject suffering from a disease or condition to the sequence of the same one or more genes of the healthy subject. Suitably to identify PTC mutations within the one or more genes of the subject suffering from a disease or condition. Suitably such sequence comparisons may be carried out by available software, such as alignment software.

[0083] PTC read-through agents, such as the compounds disclosed herein, may in one or more embodiments, be of value and used in the treatment or amelioration of the following non-limiting examples of disorders and diseases. Suitably any of the following disorders and diseases may be regarded as a disease or disorder associated with a PTC mutation in a subject. Suitably therefore the present invention provides a method of treating conditions or disorders which are associated with PTC mutations in a subject, wherein the conditions or disorders are selected from any listed in the following paragraphs.

[0084] The disease may be a disease of the musculoskeletal system. The disease of the musculoskeletal system may be a disease selected from Shwachman-Diamond Syndrome, Rickets, Laron Syndrome, Muscular Dystrophies (e.g. Duchenne muscular dystrophy(DMD)), Microcephaly, congenital limb deformities, Muscle Spasticity, Dwarfism, Gigantism, Osteopoikilosis, Cleidocranial Dysplasia, Synostosis, Mitochondrial Myopathies and Encephalomyopathies, Craniosynostoses, Mandibulofacial Dysostosis, Scoliosis, Osteoporosis, Osteopetrosis, Hyperostosis, Osteosclerosis, Osteogenesis Imperfecta, Holoprosencephaly, Acromegaly, Arthrogryposis, and Campomelic Dysplasia.

[0085] The disease may be a disease of the skin. The disease of the skin may be a disease selected from Ectodermal Dysplasia, Epidermolysis Bullosa, Focal Dermal Hypoplasia, Ichthyosis Vulgaris, Autosomal Recessive Congenital Ichthyosis (ARCI), Recessive X-linked Ichthyosis, Lamellar Ichthyosis, Congenital Ichthyosiform Erythroderma, Harlequin Ichthyosis, Epidermolytic Ichthyosis, Superficial Epidermolytic Ichthyosis, CHILD syndrome, Netherton syndrome, MEDNIK syndrome, Neutral lipid storage disease with ichthyosis, Atopic Dermatitis, Alopecia, Atrichia with papular lesions, Hypotrichosis, Incontinentia Pigmenti, Epidermolysis Bullosa Simplex, Lipodystrophy, Hidradenitis Suppurativa, Hyperpigmentation, Junctional Epidermolysis Bullosa, Oculocutaneous Albinism, Chronic Mucocutaneous Candidiasis, Chronic Mucocutaneous Ichthyosis, X-Linked Hypopigmentation, Cutis Laxa, Menkes Kinky Hair Syndrome, Palmoplantar Keratoderma, Ehlers-Danlos Syndrome, Congenital Monilethrix, Onychomycosis, Benign Familial Pemphigus, Albinism; Kindler syndrome, Cowden Syndrome, Xeroderma pigmentosum, epidermodysplasia verruciformis, Lipoid proteinosis, dyschromatosis symmetrica hereditaria, Striate keratodermas, Autosomal Recessive Congenital Anonychia, Focal dermal hypoplasia, and Skin fragility / woolly hair syndrome.

[0086] The disease may be a metabolic disease. The metabolic disease may be selected from Congenital Adrenal Hyperplasia, Inborn errors of Amino Acid Metabolism, Hypertriglyceridemia, Diabetes Mellitus, Type 1 and Type 2, Glycogen Storage Disease, Optic Atrophy, Calcinosis, Multiple Sulfatase Deficiency Disease, Fabry Disease, Hyperinsulinism, Familial Hypophosphatemia, Pseudohypoaldosteronism, Tangier Disease, Amyotrophic Lateral Sclerosis, Lactic Acidosis, Familial Amyloidosis, Mucopolysaccharidosis, Anemia, Sandhoff Disease, Cytochrome-c Oxidase Deficiency, Pyruvate Dehydrogenase Complex Deficiency Disease, Hypoglycemia, Neuronal Ceroid- Lipofuscinoses, Severe Combined Immunodeficiency, , Chronic Idiopathic Jaundice, Progeria, Hyponatremia, Wasting Syndrome, Cystinuria, Glycogen Storage Disease, Gaucher Disease, Hypolipoproteinemias, Oculocerebrorenal Syndrome, Smith-Lemli-Opitz Syndrome, Adrenoleukodystrophy, Ataxia Telangiectasia, Canavan Disease, Carbamoyl- Phosphate Synthase I Deficiency Disease, beta-Mannosidosis, and lyosomal storage diseases, wherein the lyosomal storage disease may be selected from Niemann-PickDisease Mucopolysaccharidosis type 1 (Hurler syndrome), Mucopolysaccharidosis type 6, Mucopolysaccharidosis type 7, CLN1 disease, and CL3 disease.

[0087] The disease may be a disease of the Nervous System. The disease of the Nervous System may be selected from Rett Syndrome, Ataxias, Sensorineural Hearing Loss, Epilepsy, Charcot-Marie-Tooth Disease, Spinal Muscular Atrophy, Spastic Paraplegia, Hydrocephalus, Migraine with Aura, Chorea, Tremor, Usher Syndromes, De Lange Syndrome, Duane Retraction Syndrome, Dementia, Myoclonus, Hereditary Sensory and Autonomic Neuropathies, Intellectual Disability, X-linked Mental Retardation, Fragile X Syndrome, Hypopituitarism, Leukoencephalopathies, Dystonia, Congenital Pain Insensitivity, Tourette Syndrome, Alzheimers Disease, Parkinsons Disease, Angelman Syndrome, Apraxias, Cerebral Palsy, and Frontotemporal Dementia.

[0088] The disease may be a Cardiovascular disease. The cardiovascular disease may be selected from Coronary Disease, Ventricular Fibrillation, Telangiectasis, Kartagener Syndrome, Alagille Syndrome, Andersen Syndrome, Atrioventricular Block, and Cardiomyopathies.

[0089] The disorder may be an Endocrine disorder. The Endocrine disorder may be selected from Hypogonadism, Goiter, Fetal Macrosomia, Thyroid Hormone Resistance Syndrome, Gonadal Dysgenesis, Hypoparathyroidism, Neurogenic Diabetes Insipidus, and Androgen-Insensitivity Syndrome.

[0090] The disease may be a disease of the eye. The disease of the eye may be selected from Leber syndrome, Hereditary Optic Atrophies, Ectopia Lentis, Coloboma, Aphakia, and Choroideremia.

[0091] The disease may be a disease affecting the urogenital system. The disease affecting the urogenital system may be selected from Hypospadias, Hydrops Fetalis, Interstitial Nephritis, Polycystic Kidney Diseases, Alport Syndrome, and Azoospermia.

[0092] The disease or disorder may be a haemic or lymphatic condition. The haemic or lymphatic condition may be selected from alpha- and beta-Thalassemia, Afibrinogenemia, Hemophagocytic Lymphohistiocytosis, Factor XI Deficiency, Hemophilia A, von Willebrand Diseases, Factor V Deficiency, Sideroblastic Anemia, Hereditary Elliptocytosis, Neutropenia, Chronic Granulomatous Disease, Hereditary Spherocytosis, Polycythemia, Hemophilia B, Factor VII Deficiency, Bernard-Soulier Syndrome, Dyserythropoietic Anemia, Hemolytic Anemia, Idiopathic Thrombocytopenic Purpura, Thrombasthenia, Factor XIII Deficiency, Hepatoerythropoietic Porphyria, and Acute Intermittent Porphyria.

[0093] The disease may be a respiratory disease. The respiratory disease may be selected from Cystic Fibrosis, Pulmonary Hypertension, Lipoid Proteinosis of Urbach and Wiethe, Newborn Respiratory Distress Syndrome, Chronic Obstructive Pulmonary Disease, Chronic Obstructive, Asthma, and Choanal Atresia.

[0094] The disease or disorder may be an inflammatory or autoimmune condition. The inflammatory or autoimmune condition may be selected from Immunologic Deficiency Syndromes, and Leukocyte-Adhesion Deficiency Syndrome.

[0095] The disease may be a disease of the Gastrointestinal system. Diseases of the gastrointestinal system may be selected from Chronic Hepatitis B, Colitis, Intestinal Polyposis, Inflammatory Bowel Diseases, Hirschsprung Disease, Exocrine Pancreatic Insufficiency, Crohn’s Disease, and Cholestasis.

[0096] The disease or disorder may be a Neoplasm. The Neoplasm may be selected from Acute Myeloid Leukemia, Paraganglioma, Rhabdoid Tumor, Rhabdomyoma, Adenoid Cystic Carcinoma, Large Cell Carcinoma, Lobular Carcinoma, Skin Appendage Carcinoma, Squamous Cell Carcinoma, Alveolar Rhabdomyosarcoma, Neuroectodermal Tumors, Multiple Hamartoma Syndrome, Pheochromocytoma, Nevus, Osteosarcoma, Teratoma, and Adenoma.

[0097] The disease or disorder may be cancer. The cancer may be selected from acoustic neuroma, anal cancer, bladder cancer, Bowen's disease, brain cancer, breast cancer, carcinomas including basal cell carcinoma, bile duct carcinoma, bronchogenic carcinoma, choriocarcinoma, embryonal carcinoma, cystadenocarcinoma, epithelial carcinoma, medullary carcinoma, NUT midline carcinoma (NMC), papillary carcinoma, papillary adenocarcinomas, renal cell carcinoma, sebaceous gland carcinoma, small cell lung carcinoma, squamous cell carcinoma, and sweat gland carcinoma, cervical cancer, chordoma, colon cancer, colorectal cancer, craniopharyngioma, dysproliferative changes (dysplasias and metaplasias), endometrial cancer, ependymoma, esophageal cancer, essential thrombocythemia, estrogen-receptor positive breast cancer, Ewing’s tumour, genital cancer, cancer of the cervix, cancer of the vulva, vulvar intraepithelial neoplasia (VIN), cancer of the vagina, germ cell testicular cancer, gastrointestinal cancers, gastric cancer, glioblastoma, glioma, heavy chain disease, hemangioblastoma, hepatocellular cancer, hepatoma, hormone insensitive prostate cancer, keratinocyte carcinomas, kidney cancer, leukaemias including acute leukaemia, acute lymphocytic leukaemia, acute myeloid leukaemia, acute myelocytic leukaemia (monocyctic, myeloblastic, adenocarcinoma, angiosarcoma, astrocytoma, myelomonocytic and promyelocytic), acute t-cell leukaemia, chronic leukaemia, chronic lymphocytic leukaemia, chronic myelocytic (granulocytic)leukaemia, chronic myelogenous leukaemia, erythroleukemia, lymphoblastic leukaemia, and myelogenous leukaemia, liver cancer, lung cancer, lymphoid malignancies of T-cell or B-cell origin, lymphomas (Hodgkin’s and non-Hodgkin’s) including cutaneous T-cell lymphoma, diffuse large B-cell lymphoma, and follicular lymphoma, cutaneous (skin) lymphomas, malignancies and hyperproliferative disorders including of the bladder, breast, colon, lung, ovaries, pancreas, prostate, skin and uterus, advanced malignancies, medulloblastoma, melanoma, meningioma, Merkel cell cancer mesothelioma, metastatic cancer, multiple myeloma, myeloma, pancreatic cancer, myelofibrosis, myeloproliferative neoplasms, neuroblastoma, non-small cell lung cancer, head and neck cancer, oligodendroglioma, oral cancer, ovarian cancer, pancreatic cancer, pinealoma, polycythemia vera, prostate cancer, rectal cancer, retinoblastoma, sarcomas including chondrosarcoma, endotheliosarcoma, fibrosarcoma, gliosarcoma, leiomyosarcoma, liposarcoma, lymphagioendotheliosarcoma, lymphangiosarcoma, myxosarcoma, Castleman's disease and Kaposi's sarcoma, osteogenic sarcoma, and rhabdomyosarcoma, seminoma, skin cancer, skin adnexal tumors, and sarcomas, small cell lung cancer, solid tumors, stomach cancer, synovioma, testicular tumours, thyroid cancer, uterine cancer, Waldenstrom’s macroglobulinemia, and Wilms’ tumour.

[0098] The disease or disorder may be selected from Amelogenesis Imperfecta, Anodontia, Odontodysplasia, Branchio-Oto-Renal Syndrome, Sotos Syndrome, and Waardenburg's Syndrome.

[0099] The disease or disorder may be selected from: Recessive dystrophic epidermolysis bullosa, Junctional epidermolysis bullosa, Xeroderma pigmentosum, Netherton syndrome, Duchenne Muscular Dystrophy, Becker Muscular Dystrophy, Cystic Fibrosis, Alport Syndrome, Dravet Syndrome, Aniridia, Methylmalonic Acidemia, Colorectal Cancer, Endometrium Cancer, Breast Cancer, Ovarian Cancer, Lung Squamous Cell Carcinoma, Head and Neck Squamous Cell Carcinoma, Familial adenomatous polyposis, Hemophilia A, Hemophilia B, Choroideremia, Pulmonary Artery Hypertension, Ataxia telangiectasia, Shwachman-Diamond syndrome, Mucopolysaccharidosis Type I, Mucopolysaccharidosis Type VI, Mucopolysaccharidosis type III, Niemann-Pick Disease, Primary Ciliary Dyskinesia, and ciliopathies (such as Usher syndrome or Retinitis Pigmentosa).

[0100] The disease or disorder may be selected from: Recessive dystrophic epidermolysis bullosa, Junctional epidermolysis bullosa, Duchenne Muscular Dystrophy, Cystic Fibrosis, Colorectal Cancer, Endometrium Cancer, Breast Cancer, Ovarian Cancer, Lung Squamous Cell Carcinoma, Head and Neck Squamous Cell Carcinoma, and Familial adenomatous polyposis. Suitably such disorders may be associated with a PTC mutation in a subject.Suitably for example, the disease may be Junctional epidermolysis bullosa, associated with a PTC mutation in the LAMB3 gene. Suitably for example, the disease may be Recessive dystrophic epidermolysis bullosa, associated with a PTC mutation in the COL.7A1 gene. Suitably for example, the disease may be Duchenne Muscular Dystrophy, associated with a PTC mutation in the DMD gene. Suitably for example, the disease may be Cystic Fibrosis associated with a PTC mutation in the CTFR gene. Suitably for example, the disease may be Familial adenomatous polyposis associated with a PTC mutation in the APC gene. Suitably for example, the disease may be a cancer as listed above associated with a PTC mutation in a gene selected from: TP53, PTEN, ARID1A or CTCF.

[0101] Suitably, therefore, the present invention provides a method of treating conditions or disorders which are associated with PTC mutations in a subject, wherein the condition or disorder associated with PTC mutations in a subject is selected from recessive dystrophic epidermolysis bullosa, Junctional epidermolysis bullosa, Xeroderma pigmentosum, Netherton syndrome, Duchenne Muscular Dystrophy, Becker Muscular Dystrophy, Cystic Fibrosis, Dravet Syndrome, Aniridia, Methylmalonic Acidemia, Familial adenomatous polyposis, Hemophilia A, Hemophilia B, Choroideremia, Pulmonary Artery Hypertension, Ataxia telangiectasia, Shwachman-Diamond syndrome, Mucopolysaccharidosis Type I, Mucopolysaccharidosis Type VI, Mucopolysaccharidosis type III, Niemann-Pick Disease, Primary Ciliary Dyskinesia, Ciliopathies (such as Usher syndrome or Retinitis Pigmentosa) and Alport Syndrome. In some embodiments, the present invention provides a method of treating conditions or disorders which are associated with PTC mutations in a subject, wherein the condition or disorder associated with PTC mutations in a subject is selected from recessive dystrophic epidermolysis bullosa, Junctional epidermolysis bullosa, Duchenne Muscular Dystrophy, Cystic Fibrosis, and Familial adenomatous polyposis.

[0102] PTC read-through agents, such as the compounds disclosed herein, may in one or more embodiments, also be of value and used in the palliation, diagnosis or prevention of any disease, disorder or condition in humans of one or more of the aforesaid non-limiting examples of disorders and diseases. Suitably any of the aforesaid disorders and diseases may be regarded as a disease or disorder associated with a PTC mutation in a subject. Suitably therefore the present invention further provides a method of palliation, diagnosis or prevention of conditions or disorders which are associated with PTC mutations in a subject, wherein the conditions or disorders are selected from any listed in the aforesaid paragraphs.

[0103] Treatment or amelioration with PTC read-through agents, such as compositions comprising the compounds disclosed herein or salts thereof (or combinations thereof), in some embodiments may be effective if administered orally. In some other embodiments maybe effective if applied topically, and in some further embodiments may be effective if applied topically and orally.

[0104] In some embodiments, compositions comprising a novel compound disclosed herein or salt thereof (or combinations thereof) may be administered to young children. In some embodiments, compositions comprising a compound of the invention or salt thereof (or combinations thereof) may be administered to adolescents or teenagers. In some embodiments, compositions comprising a compound of the invention or salt thereof (or combinations thereof) may be administered to adults.BRIEF DESCRIPTION OF THE DRAWINGS

[0105] Embodiments of the invention are further described hereinafter with reference to the accompanying drawings, in which:Figure 1 shows the effect of treatment of HDQ-P1 cells with Example 1 (1.23 pM, 3.7 pM, 11 pM, or 33 pM) compared to DMSO (0.3%), and G418 (200 pM).DETAILED DESCRIPTION

[0106] The term Cm-Cnrefers to a group with m to n carbon atoms. For the absence of doubt, the term “Co” refers to a group with 0 carbon atoms.

[0107] The term “alkyl” refers to a monovalent linear or branched saturated hydrocarbon chain. For example, Ci-Ce-alkyl may refer to methyl, ethyl, n-propyl, / so-propyl, n-butyl, secbutyl, tert-butyl, n-pentyl and n-hexyl. The alkyl groups may be unsubstituted or substituted by one or more substituents.

[0108] The term “alkylene” refers to a bivalent linear saturated hydrocarbon chain. For example, Ci-Cs-alkylene may refer to methylene, ethylene or propylene. The alkylene groups may be unsubstituted or substituted by one or more substituents. For the absence of doubt, the term “Co-alkylene” refers to a group in which an alkylene chain is absent. For example, “Co-alkylene-Ra” refers to an Ra.

[0109] The term “haloalkyl” refers to a hydrocarbon chain substituted with at least one halogen atom independently chosen at each occurrence from: fluorine, chlorine, bromine and iodine. The halogen atom may be present at any position on the hydrocarbon chain. For example, Ci-Ce-haloalkyl may refer to chloromethyl, fluoromethyl, trifluoromethyl, chloroethyl e.g. 1 -chloromethyl and 2-chloroethyl, trichloroethyl e.g. 1 ,2,2-trichloroethyl, 2,2,2- trichloroethyl, fluoroethyl e.g. 1 -fluoromethyl and 2-fluoroethyl, trifluoroethyl e.g. 1 ,2,2-trifluoroethyl and 2,2,2-trifluoroethyl, chloropropyl, trichloropropyl, fluoropropyl, trifluoropropyl. A haloalkyl group may be a fluoroalkyl group, i.e. a hydrocarbon chain substituted with at least one fluorine atom. Thus, a haloalkyl group may have any amount of halogen substituents. The group may contain a single halogen substituent, it may have two or three halogen substituents, or it may be saturated with halogen substituents.

[0110] The term “alkenyl” refers to a branched or linear hydrocarbon chain containing at least one double bond. The double bond(s) may be present as the E or Z isomer. The double bond may be at any possible position of the hydrocarbon chain. For example, “C2-C6-alkenyl” may refer to ethenyl, propenyl, butenyl, butadienyl, pentenyl, pentadienyl, hexenyl and hexadienyl. The alkenyl groups may be unsubstituted or substituted by one or more substituents.

[0111] The term “alkynyl” refers to a branched or linear hydrocarbon chain containing at least one triple bond. The triple bond may be at any possible position of the hydrocarbon chain. For example, “C2-C6-alkynyl” may refer to ethynyl, propynyl, butynyl, pentynyl and hexynyl. The alkynyl groups may be unsubstituted or substituted by one or more substituents.

[0112] The term “cycloalkyl” refers to a saturated hydrocarbon ring system containing 3, 4, 5 or 6 carbon atoms. For example, “Cs-Ce-cycloalkyl” may refer to cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl. The cycloalkyl groups may be unsubstituted or substituted by one or more substituents.

[0113] The term “y- to z-membered heterocycloalkyl” refers to a y- to z- membered heterocycloalkyl group. Thus it may refer to a monocyclic or bicyclic saturated or partially saturated group having from y to z atoms in the ring system and comprising 1 or 2 heteroatoms independently selected from O, S and N in the ring system (in other words 1 or 2 of the atoms forming the ring system are selected from O, S and N). By partially saturated it is meant that the ring may comprise one or two double bonds. This applies particularly to monocyclic rings with from 5 to 6 members. The double bond will typically be between two carbon atoms but may be between a carbon atom and a nitrogen atom. Examples of heterocycloalkyl groups include; piperidine, piperazine, morpholine, thiomorpholine, pyrrolidine, tetra hydrofuran, tetrahydrothiophene, dihydrofuran, tetrahydropyran, dihydropyran, dioxane, azepine. A heterocycloalkyl group may be unsubstituted or substituted by one or more substituents.

[0114] Aryl groups may be any aromatic carbocyclic ring system (i.e. a ring system containing 2(2n + 1)TT electrons). Aryl groups may have from 6 to 10 carbon atoms in the ring system. Aryl groups will typically be phenyl groups. Aryl groups may be naphthyl groups or biphenyl groups.

[0115] The term ‘heterocyclyl’ group refers to rings comprising from 1 to 4 heteroatoms independently selected from O, S and N. The rings may be heterocycloalkyl rings (including both saturated and partially saturated rings) or heteroaryl rings. The term “heterocyclyl” also encompasses groups that are tautomers of hydroxy heteroaryl groups, such pyridones, and tautomers of hydroxy heteroaryl groups that are substituted on the nitrogen, such as N-alkyl pyridones.

[0116] The term ‘heterocycloalkenyl’ refers to partially saturated rings comprising from 1 to 2 heteroatoms independently selected from O, S and N.

[0117] The term “heteroaryl” refers to any aromatic (i.e. a ring system containing 2(2n +1 )TT electrons) 5, 6-, 8-, 9- or 10-membered ring system comprising from 1 to 4 heteroatoms independently selected from O, S and N (in other words from 1 to 4 of the atoms forming the ring system are selected from O, S and N). Thus, any heteroaryl groups may be independently selected from: 5 membered heteroaryl groups in which the heteroaromatic ring is substituted with 1-4 heteroatoms independently selected from O, S and N; and 6- membered heteroaryl groups in which the heteroaromatic ring is substituted with 1-3 (e.g.1-2) nitrogen atoms. Heterocyclic groups may be bicyclic. Thus they might be 6,6 bicyclic systems (e.g. quinazolines, quinoline, isoquinoline), 5,6 bicyclic systems (e.g. indoles, benzimidazoles, benzopyrazoles) or 5,5-bicyclic systems (e.g. dihydropyrrolopyrroles). Specifically, heteroaryl groups may be independently selected from: pyrrole, furan, thiophene, pyrazole, imidazole, oxazole, isoxazole, triazole, oxadiazole, thiadiazole, tetrazole; pyridine, pyridazine, pyrimidine, pyrazine, triazine. .

[0118] Compounds of the invention containing one or more asymmetric carbon atoms can exist as two or more stereoisomers. Where a compound of the invention contains a double bond such as a C=C or C=N group, geometric cis / trans (or Z / E) isomers are possible. Where structural isomers are interconvertible via a low energy barrier, tautomeric isomerism (‘tautomerism’) can occur. This can take the form of proton tautomerism in compounds of the invention containing, for example, an imino, keto, or oxime group, or so-called valence tautomerism in compounds which contain an aromatic moiety. It follows that a single compound may exhibit more than one type of isomerism.

[0119] Included within the scope of the present invention are all stereoisomers, geometric isomers and tautomeric forms of the compounds of the invention, including compounds exhibiting more than one type of isomerism, and mixtures of one or more thereof. Also included are acid addition or base salts wherein the counter ion is optically active, for example, d-lactate or l-lysine, or racemic, for example, dl-tartrate or dl-arginine.

[0120] Cis / trans isomers may be separated by conventional techniques well known to those skilled in the art, for example, chromatography and fractional crystallisation.

[0121] Conventional techniques for the preparation / isolation of individual enantiomers when necessary include chiral synthesis from a suitable optically pure precursor or resolution of the racemate (or the racemate of a salt or derivative) using, for example, chiral high pressure liquid chromatography (HPLC). Thus, chiral compounds of the invention (and chiral precursors thereof) may be obtained in enantiomerically-enriched form using chromatography, typically HPLC, on an asymmetric resin with a mobile phase consisting of a hydrocarbon, typically heptane or hexane, containing from 0 to 50% by volume of isopropanol, typically from 2% to 20%, and for specific examples, 0 to 5% by volume of an alkylamine e.g. 0.1% diethylamine. Concentration of the eluate affords the enriched mixture.

[0122] Alternatively, the racemate (or a racemic precursor) may be reacted with a suitable optically active compound, for example, an alcohol, or, in the case where the compound of the invention contains an acidic or basic moiety, a base or acid such as 1 -phenylethylamine or tartaric acid. The resulting diastereomeric mixture may be separated by chromatography and / or fractional crystallization and one or both of the diastereoisomers converted to the corresponding pure enantiomer(s) by means well known to a skilled person.

[0123] When any racemate crystallises, crystals of two different types are possible. The first type is the racemic compound (true racemate) referred to above wherein one homogeneous form of crystal is produced containing both enantiomers in equimolar amounts. The second type is the racemic mixture or conglomerate wherein two forms of crystal are produced in equimolar amounts each comprising a single enantiomer.

[0124] While both of the crystal forms present in a racemic mixture have identical physical properties, they may have different physical properties compared to the true racemate. Racemic mixtures may be separated by conventional techniques known to those skilled in the art - see, for example, “Stereochemistry of Organic Compounds” by E. L. Eliel and S. H. Wilen (Wiley, 1994).

[0125] Suitable pharmaceutically acceptable salts include, but are not limited to, salts of pharmaceutically acceptable inorganic acids such as hydrochloric, sulphuric, phosphoric, nitric, carbonic, boric, sulfamic, and hydrobromic acids, or salts of pharmaceutically acceptable organic acids such as acetic, propionic, butyric, tartaric, maleic, hydroxymaleic, fumaric, malic, citric, lactic, mucic, gluconic, benzoic, succinic, oxalic, phenylacetic, methanesulphonic, toluenesulphonic, benzenesulphonic, salicylic, sulphanilic, aspartic, glutamic, edetic, stearic, palmitic, oleic, lauric, pantothenic, tannic, ascorbic and valeric acids. Suitable base salts are formed from bases which form non-toxic salts. Examples include thealuminium, arginine, benzathine, calcium, choline, diethylamine, diolamine, glycine, lysine, magnesium, meglumine, olamine, potassium, sodium, tromethamine and zinc salts. Hemisalts of acids and bases may also be formed, for example, hemisulfate and hemicalcium salts.

[0126] Compounds and salts described in this specification may be isotopically-labelled (or “radio-labelled”). Accordingly, one or more atoms are replaced by an atom having an atomic mass or mass number different from the atomic mass or mass number typically found in nature. Examples of radionuclides that may be incorporated include2H (also written as “D” for deuterium),3H (also written as “T” for tritium),11C,13C,14C,15O,17O,18O,13N,15N,18F,36CI,123l,25l,32P,35S and the like. The radionuclide that is used will depend on the specific application of that radio-labelled derivative. For example, for in vitro competition assays,3H or14C are often useful. For radio-imaging applications,11C or18F are often useful. In some embodiments, the radionuclide is3H. In some embodiments, the radionuclide is14C. In some embodiments, the radionuclide is11C. And in some embodiments, the radionuclide is18F.

[0127] Isotopically-labelled compounds can generally be prepared by conventional techniques known to those skilled in the art or by processes analogous to those described using an appropriate isotopically-labelled reagent in place of the non-labelled reagent previously employed.

[0128] The selective replacement of hydrogen with deuterium in a compound may modulate the metabolism of the compound, the PK / PD properties of the compound and / or the toxicity of the compound. For example, deuteration may increase the half-life or reduce the clearance of the compound in vivo. Deuteration may also inhibit the formation of toxic metabolites, thereby improving safety and tolerability. It is to be understood that the invention encompasses deuterated derivatives of compounds of formula (I). As used herein, the term deuterated derivative refers to compounds of the invention where in a particular position at least one hydrogen atom is replaced by deuterium. Accordingly, in a compound of the invention one or more hydrogen atom is optionally replaced by deuterium. For example, one or more hydrogen atoms in a Ci-4-alkyl group may be replaced by deuterium to form a deuterated Ci-4-alkyl group. By way of example, if any of R2, R3, R4, R5, R6, R7, R7a, R8, R9, Raor Rbis methyl, the invention also encompasses -CDs, -CHD2 and -CH2D. Similarly R2, R3, R4, R7, R7a, R9, Ra, or Rbmay be D.

[0129] The activity of the compounds of the present invention can be assessed by a variety of in silico, in vitro and in vivo assays. In silico analysis of a variety of compounds has been demonstrated to be predictive of ultimate in vitro and even in vivo activity.

[0130] It is to be appreciated that references to “treating” or “treatment” include prophylaxis as well as the alleviation of established symptoms of a condition. “Treating” or “treatment” of a state, disorder or condition therefore includes: (1) preventing or delaying the appearance of clinical symptoms of the state, disorder or condition developing in a human that may be afflicted with or predisposed to the state, disorder or condition but does not yet experience or display clinical or subclinical symptoms of the state, disorder or condition, (2) inhibiting the state, disorder or condition, i.e., arresting, reducing or delaying the development of the disease or a relapse thereof (in case of maintenance treatment) or at least one clinical or subclinical symptom thereof, or (3) relieving or attenuating the disease, i.e., causing regression of the state, disorder or condition or at least one of its clinical or subclinical symptoms.

[0131] A “therapeutically effective amount” includes the amount of a compound that, when administered to a mammal for treating a disease, is sufficient to affect such treatment for the disease. The "therapeutically effective amount" will vary depending on the compound, the disease and its severity and the age, weight, etc., of the mammal to be treated.

[0132] A compound of the invention, or pharmaceutically acceptable salt thereof, may be used on their own but will generally be administered in the form of a pharmaceutical composition in which the compounds of the invention, or pharmaceutically acceptable salt thereof, is in association with a pharmaceutically acceptable adjuvant, diluent or carrier.

[0133] Conventional procedures for the selection and preparation of suitable pharmaceutical formulations are described in, for example, "Pharmaceuticals - The Science of Dosage Form Designs", M. E. Aulton, Churchill Livingstone, 1988.

[0134] Depending on the mode of administration of the compounds of the invention, the pharmaceutical composition which is used to administer the compounds of the invention will preferably comprise from 0.05 to 99 % w / w compounds of the invention, more preferably from 0.05 to 80 % w / w compounds of the invention, still more preferably from 0.10 to 70 % w / w compounds of the invention, and even more preferably from 0.10 to 50 % w / w compounds of the invention (all percentages by weight being based on total composition).

[0135] The pharmaceutical compositions may be administered topically (e.g. to the skin) in the form, e.g., of creams, ointments, gels, lotions, solutions, suspensions; or systemically, e.g. by oral administration in the form of tablets, lozenges, hard or soft capsules, aqueous or oily suspensions, emulsions, dispersible powders or granules, syrups or elixirs; or by parenteral administration in the form of a sterile aqueous or oily solution, suspension or emulsion for injection (including intravenous, intracoronary, subcutaneous, intramyocardial, intraperitoneal, intramuscular, intravascular or infusion); by rectal administration in the formof suppositories or enemas; by inhalation for example as a finely divided powder or a liquid aerosol; or for administration by insufflation (for example as a finely divided powder).

[0136] For oral administration the compounds of the invention may be admixed with an adjuvant or a carrier, for example, lactose, saccharose, sorbitol, mannitol; a starch, for example, potato starch, corn starch or amylopectin; a cellulose derivative; a binder, for example, gelatine or polyvinylpyrrolidone; and / or a lubricant, for example, magnesium stearate, calcium stearate, polyethylene glycol, a wax, paraffin, and the like, and then compressed into tablets. If coated tablets are required, the cores, prepared as described above, may be coated with a concentrated sugar solution which may contain, for example, gum arabic, gelatine, talcum and titanium dioxide. Alternatively, the tablet may be coated with a suitable polymer dissolved in a readily volatile organic solvent. Thus, compositions intended for oral use may contain, for example, one or more colouring, sweetening, flavouring and / or preservative agents.

[0137] For the preparation of soft gelatine capsules, the compounds of the invention may be admixed with, for example, a vegetable oil or polyethylene glycol. Hard gelatine capsules may contain granules of the compound using either the above-mentioned excipients for tablets. Also liquid or semisolid formulations of the compound of the invention may be filled into hard gelatine capsules. Liquid preparations for oral application may be in the form of syrups or suspensions, for example, solutions containing the compound of the invention, the balance being sugar and a mixture of ethanol, water, glycerol and propylene glycol. Optionally such liquid preparations may contain colouring agents, flavouring agents, sweetening agents (such as saccharine), preservative agents and / or carboxymethylcellulose as a thickening agent or other excipients known to those skilled in art.

[0138] For intravenous (parenteral) administration the compounds of the invention may be administered as a sterile aqueous or oily solution.

[0139] The size of the dose for therapeutic or prophylactic purposes of a compound of the invention will naturally vary according to the nature and severity of the conditions, the concentration of the compound required for effectiveness in isolated cells, the concentration of the compound required for effectiveness in experimental animals, the age and sex of the animal or patient and the route of administration, according to well known principles of medicine.

[0140] Dosage levels, dose frequency, and treatment durations of compounds of the invention are expected to differ depending on the formulation and clinical indication, age, and co-morbid medical conditions of the patient.

[0141] An effective amount of a compound of the present invention for use in therapy of a condition is an amount sufficient to achieve symptomatic relief in a warm-blooded animal, particularly a human of the symptoms of the condition, to mitigate the physical manifestations of the condition, or to slow the progression of the condition.

[0142] The amount of active ingredient that is combined with one or more excipients to produce a single dosage form will necessarily vary depending upon the host treated and the particular route of administration. For example, a formulation intended for oral administration to humans will generally contain, for example, from 0.5 mg to 0.5 g of active agent (more suitably from 0.5 to 100 mg, for example from 1 to 30 mg) compounded with an appropriate and convenient amount of excipients which may vary from about 5 to about 98 percent by weight of the total composition.

[0143] For the above-mentioned compounds of the invention the dosage administered will, of course, vary with the compound employed, the mode of administration, the treatment desired and the disorder indicated. In using a compound of the invention for therapeutic or prophylactic purposes it will generally be administered so that a daily dose in the range, for example, a daily dose selected from 0.1 mg / kg to 100 mg / kg, 1 mg / kg to 75mg / kg, 1 mg / kg to 50 mg / kg, 1 mg / kg to 20 mg / kg or 5 mg / kg to 10 mg / kg body weight is received, given if required in divided doses. In general lower doses will be administered when a parenteral route is employed. Thus, for example, for intravenous or intraperitoneal administration, a dose in the range, for example, 0.1 mg / kg to 30 mg / kg body weight will generally be used. Similarly, for administration by inhalation, a dose in the range, for example, 0.05 mg / kg to 25 mg / kg body weight will be used. Suitably the compound of the invention is admistered orally, for example in the form of a tablet, or capsule doasage form. The daily dose administered orally may be, for example a total daily dose selected from 1 mg to 1000 mg, 5 mg to 1000 mg, 10 mg to 750 mg or 25 mg to 500 mg. Typically, unit dosage forms will contain about 0.5 mg to 0.5 g of a compound of this invention.

[0144] The compounds of the invention may be administered along with other active compounds as part of a treatment regime. The other active compounds may be administered simultaneously with, subsequently to or previously to the administration of the compounds of the invention. It may be that the pharmaceutical formulation comprising the compounds of the invention also comprises one or more other active compounds. The other active compounds may be anticancer, anti-inflammatory, antibacterial, antiviral, antiemetic, antithrombotic or compounds that alter the metabolism.

[0145] The other active compound may be an aminoglycoside. The other active compound may be an eRF1 modulator. The other active compound may be an inhibitor or suppressorof nonsense mediated decay. The other active compound may be an SMG1 kinase inhibitor. The other active compound may be a ribosomal binder.

[0146] The compounds of the invention may be administered in combination with an aminoglycoside. In some embodiments, the compounds of the invention may allow translational read-through of PTC mutations when administered in combination with a subeffective, sub-optimal, or sub-maximal amount of an aminoglycoside.

[0147] Aminoglycosides are compounds that have an amino-modified glycoside. Aminoglycosides are compounds that have an amino-modified glycoside. Aminoglycosides may allow translational read-through of PTC mutations. In some embodiments, when a compound of the invention is administered in combination with an aminoglycoside, the aminoglycoside may increase ribosomal read-through of mRNA transcripts carrying a PTC mutation of the cystic fibrosis CFTR channel.

[0148] Non-limiting examples of aminoglycosides include kanamycin A, amikacin, tobramycin, dibekacin, gentamicin, geneticin, sisomicin, netilmicin, neomycin B, neomycin C, paromomycin, streptomycin, plazomicin, tobramycin, ELX-02, and pharmaceutically acceptable salts thereof.

[0149] It may be that administering a compound of the invention in combination with an aminoglycoside enhances PTC read-through. It may be that administering a compound of the invention in combination with an aminoglycoside enhances PTC read-through whilst reducing the dose of aminoglycoside administered, relative to administration of the aminoglycoside alone, thereby reducing side effects, e.g. toxicity, associated with the aminoglycoside.

[0150] It may be that administering a compound of the invention in combination with an aminoglycoside increases the potency and / or beneficial effects of the aminoglycoside.

[0151] It may be that administering a compound of the invention in combination with an aminoglycoside reduces the dose of aminoglycoside required by 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or more, compared to administration of the aminoglycoside alone.

[0152] The compounds of the invention may be administered in combination with an eRF1 modulator, such as SRI-41315.

[0153] The compounds of the invention may be administered in combination with an inhibitor or suppressor of nonsense mediated decay, such as amlexanox, or NMD1.

[0154] The compounds of the invention may be administered in combination with an SMG1 kinase inhibitor, such as SMG1 L

[0155] The compounds of the invention may be administered in combination with an ribosomal binder, such as ZKN-013.

[0156] The compounds of the invention may be administered in combination with an antibody, for example a checkpoint inhibitor antibody. In some embodiments, the compounds of the invention may allow increased immune-cell recognition of cancer cells for use in the treatment of cancer, when administered in combination with a checkpoint inhibitor antibody. In some embodiments, the compounds of the invention may allow increased immune-cell recognition of cancer cells for use in the treatment of cancer, when administered in combination with a sub-effective, sub-optimal, or sub-maximal amount of a checkpoint inhibitor antibody.

[0157] Checkpoint inhibitor antibodies are a type of immunotherapy in oncology. Checkpoint inhibitor antibodies recognise and block different checkpoint proteins, for example, CTLA-4, PD-1 and PD-L1 , which triggers immune recognition and destruction of a tumour. Immune recognition may be enhanced when a compound of the invention is administered in combination with a checkpoint inhibitor antibody, wherein the compound promotes read-through of PTCs and increases the expression of neoantigens in cancer cells.

[0158] Non-limiting examples of checkpoint inhibitor antibodies include pembrolizumab, nivolumab, cemiplimab, atezolizumab, durvalumab, avelumab, relatlimab, and ipilimumab.

[0159] It may be that administering a compound of the invention in combination with a checkpoint inhibitor antibody increases the potency and / or beneficial effects of the checkpoint inhibitor antibody.

[0160] Throughout the description and claims of this specification, the words “comprise” and “contain” and variations of them mean “including but not limited to”, and they are not intended to (and do not) exclude other moieties, additives, components, integers or steps. Throughout the description and claims of this specification, the singular encompasses the plural unless the context otherwise requires. In particular, where the indefinite article is used, the specification is to be understood as contemplating plurality as well as singularity, unless the context requires otherwise.

[0161] Features, integers, characteristics, compounds, chemical moieties or groups described in conjunction with a particular aspect, embodiment or example of the invention are to be understood to be applicable to any other aspect, embodiment or example described herein unless incompatible therewith. All of the features disclosed in this specification (including any accompanying claims, abstract and drawings), and / or all of the steps of any method or process so disclosed, may be combined in any combination, except combinations where at least some of such features and / or steps are mutually exclusive. The invention isnot restricted to the details of any foregoing embodiments. The invention extends to any novel one, or any novel combination, of the features disclosed in this specification (including any accompanying claims, abstract and drawings), or to any novel one, or any novel combination, of the steps of any method or process so disclosed.

[0162] It will be understood that various modifications may be made to the embodiments disclosed herein. Therefore the above description should not be construed as limiting, but merely as exemplifications of preferred embodiments. For example, the functions described above and implemented as the best mode for operating the present invention are for illustration purposes only. Other arrangements and methods may be implemented by those skilled in the art without departing from the scope and spirit of this invention. Moreover, those skilled in the art will envision other modifications within the scope and spirit of the specification appended hereto.

[0163] The reader's attention is directed to all papers and documents which are filed concurrently with or previous to this specification in connection with this application and which are open to public inspection with this specification, and the contents of all such papers and documents are incorporated herein by reference.

[0164] The compounds of formula (I) can be made according to or analogously to General Schemes 1 to 3 and / or the following Examples 1 to 13.EXAMPLESTable of Abbreviations:NMR Methods:1H NMR spectra are recorded on Bruker AVANCE III HD 300, Bruker AVANCE NEO 400 or Bruker AVANCE III HD 400 spectrometers. Chemical shifts are denoted in ppm (6) relative to residual protonated solvent as an internal standard as described in, for example, Gottlieb et al. Journal of Organic Chemistry (1997) 62 7512. The splitting pattern for NMR spectraare denoted as follows: s (singlet), br (broad), d (doublet), t (triplet), m (multiplet) or combinations thereof. Coupling constants ( ) are designated in Hz and reported to one decimal place.5 Purification Methods:Liquid chromatography-mass spectra (LCMS) are recorded using the following systems and running conditions:0Purification by preparative HPLC (prep-HPLC) employs the following instruments and conditions:Process for Preparation:

[0165] Certain compounds of the invention may be synthesised according to the general methods disclosed herein. Certain compounds of the invention may be synthesised 5 according to general schemes 1 to 3. Certain compounds of the invention may be synthesised according to or analogously to the syntheses provided in example 1.General Scheme 1Intermediates A Intermediates B

[0166] Compounds of Formula (I) can be synthesised according to the General Scheme 1 and as described herein. Intermediates A can be optionally protected with a suitable protection group (for example Boc, SEM etc.) using conditions well described in the literature in, for example, Greene's Protective Groups in Organic Synthesis, Fourth Edition by P. G. M. Wuts and T. W. Greene. It will be apparent to those skilled in the art that the protecting group P may be attached to mainly one of the two imidazole ring nitrogens or give mixtures of both in varying ratios.

[0167] Reaction with a (hetero)aromatic ring with a suitable leaving group X (for example Cl, Br, OTf etc.) can be achieved using substitution conditions in the presence of a suitable base (for example potassium terf-butoxide) and suitable solvent (for example DMSO) with heating if required. Alternatively, reaction can be achieved under palladium catalysed crosscoupling conditions using a suitable catalyst (for example palladium acetate), with a suitable ligand (for example Xphos), with a suitable base (for example CS2CO3) and a suitable solvent (for example 1 ,4-dioxane) with heating if required.

[0168] Removal of the protecting group can be carried out under a variety of conditions well known to those skilled in the art and described in, for example, Greene's Protective Groups in Organic Synthesis, Fourth Edition by P. G. M. Wuts and T. W. Greene. It will be apparent to those skilled in the art that under certain conditions the protecting group P may not be required to enable the reaction.General Scheme 2

[0169] Intermediates A and B may be synthesised according to General Scheme 2 and as described herein. Intermediates C and D are commercially available or can be synthesised according to the methods described herein or as adapted from routes described in the literature in, for example, Journal of Medicinal Chemistry (2009), 52, 6297-6313; Archiv der Pharmazie (2014), 347, 77-88; Liebigs Annalen der Chemie (1992), 4, 317-23; US20080051578 A1; W02004011446 A1; Bioorganic & Medicinal Chemistry Letters (2016), 26, 5263-5266.

[0170] Diazenylation can be carried out using the methods described herein or as adapted from methods described in the literature in, for example, European Journal of Medicinal Chemistry (2017), 125, 327-334; JP2003300965 A; WO2017189539 A1; W02007025889 A2.

[0171] The reductive amination can be carried out using a suitable reducing agent (for example NaBH4, sodium triacetoxyborohydride, sodium cyanoborohydride) in a suitable solvent (for example methanol, THF, DCM) in the presence of an optional catalyst (for example acetic acid) if required, with heating or cooling as required.

[0172] A suitable protection group (for example Boc, SEM etc.) can be introduced using conditions well described in the literature in, for example, Greene's Protective Groups in Organic Synthesis, Fourth Edition by P. G. M. Wuts and T. W. Greene. It will be apparent to those skilled in the art that the protecting group P may be attached to mainly one of the two imidazole ring nitrogens or give mixtures of both in varying ratios.

[0173] Hydrogenation can be achieved under a variety of conditions well known to the skilled practitioner including, for example, in the presence of hydrogen gas under varying pressures and temperatures, in the presence of a suitable catalyst (for example PtO2 or palladium on carbon) in a suitable solvent (for example methanol, ethanol or isopropanol).

[0174] Halogenation coverts the alcohol into a suitable leaving group X (for example Cl, Br, I, OMs, OTs). It can be achieved by a variety of methods well known to those skilled in the art including, for example, by reaction with thionyl chloride in a suitable solvent (for example DCM or THF) with heating if required, or by well-known variations of the Appel reaction (Angewandte Chemie International Edition in English (1975) 14801-811), or by reaction with MsCI or TsCI in the presence of a suitable base (for example sodium carbonate, CS2CO3, TEA) in a suitable solvent (for example DCM, THF, DMF) with heating if required.

[0175] Substitution can be achieved by reacting the appropriate amine in a suitable solvent (for example EtOH, THF, DMF, water or combinations thereof) in the presence of a suitable base (for example sodium carbonate, CS2CO3, TEA) with a suitable catalyst if required (for example KI or nBu4NI) with heating if required.General Scheme 3

[0176] Illustrative examples of compounds of Formula (VIII) can be synthesised according to the General Scheme 3 and as described herein. The substitution rection employs a heteroaromatic ring with a suitable leaving group X (for example F, Cl, Br, OTf etc.) in the presence of a suitable base (for example potassium terf-butoxide) and suitable solvent (for example DMSO) with heating if required.

[0177] The reductive amination can be carried out using a suitable reducing agent (for example NaBH4, sodium triacetoxyborohydride, sodium cyanoborohydride) in a suitable solvent (for example methanol, THF, DCM) in the presence of an optional catalyst (for example acetic acid) if required, with heating or cooling as required.Preparation 1 - 2-nitro-1-((2-(trimethylsilyl)ethoxy)methyl)imidazole

[0178] To a stirred solution of 2-nitroimidazole (9.00 g, 79.6 mmol) in THF (90 mL) was added 60% w / w NaH (3.82 g, 95.5 mmol) in portions at 0 °C. The resulting mixture was stirred for 30 min at 0 °C. To the above mixture was added SEM-CI (16.9 mL, 95.5 mmol, 1.20 eq) dropwise at 0 °C. The resulting mixture was stirred for additional 4 h at room temperature. The reaction was quenched by the addition of water / ice (100 mL) at 0 °C. Theresulting mixture was extracted with EtOAc (3 x 100 mL). The combined organic layers were washed with brine (3 x 50 mL), dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with PE I EA (55:45) to afford the title compound as a light yellow solid (16.0 g, 82.6%). LCMS: MH+244.Preparation 2 - 5-bromo-2-nitro-1-((2-(trimethylsilyl)ethoxy)methyl)imidazole

[0179] To a solution of 2-nitro-1-{[2-(trimethylsilyl)ethoxy]methyl}imidazole (16.0 g, 65.8 mmol) in DM F (50 mL) and CHCh (50 mL) was added NBS (17.6 g, 98.6 mmol) for 5 min at room temperature. The resulting mixture was stirred overnight at room temperature. The reaction was quenched by the addition of water (300 mL). The resulting mixture was extracted with EtOAc (3 x 200 mL). The combined organic layers were washed with brine (3 x 60 mL), dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with PE I EA (8:1) to afford the title compound as a light yellow oil (13.0 g, 61.4%). LCMS: MIT 324.Preparation 3 - 5-bromo-1-((2-(trimethylsilyl)ethoxy)methyl)imidazol-2-amine

[0180] To a solution of 5-bromo-2-nitro-1-{[2-(trimethylsilyl)ethoxy]methyl}imidazole (13.0 g, 40.3 mmol) in EtOH (100 mL) and H2O (20 mL) at room temperature was added Fe (11.3 g, 202 mmol) and NH4CI (5.39 g, 101 mmol). The mixture was stirred overnight at room temperature. The resulting mixture was filtered. The filter cake was washed with EtOH (3 x 100 mL). The filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with PE I EA (35:65) to afford the title compound as a reddish brown solid (7.30 g, 61.9%). LCMS: MH+294.Preparation 4 - 2-chloro-4-methylquinazoline

[0181] To a solution of 3 M MeMgCI in THF (20.1 mL, 60.3 mmol) at -78 °C under nitrogen atmosphere was added ZnBr2 (13.6 g, 60.3 mmol) in THF (37.5 mL) dropwise over 5 min. The resulting mixture was stirred for additional 1 h at -78 °C. To the above mixture was added Pd(PPhs)4 (2.90 g, 2.51 mmol) and 2,4-dichloroquinazoline (10.0 g, 50.2 mmol) in THF (237 mL) at room temperature. The resulting mixture was stirred overnight at 50 °C. The mixture was allowed to cool down to room temperature. The reaction was quenched by the addition of sat. NH4CI (aq.) (100 mL) at 0 °C. The resulting mixture was extracted with EtOAc (3 x 80 mL). The combined organic layers were washed with brine (3 x 50 mL), dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, elutedwith PE I EA (4:6) to afford the title compound as a light yellow solid (4.80 g, 53.5%). LCMS: MIT 179.Preparation 5 - N-(5-bromo-1-{[2-(trimethylsilyl)ethoxy]methyl}imidazol-2-yl)-4- methylquinazolin-2-amine

[0182] To a solution of 5-bromo-1-{[2-(trimethylsilyl)ethoxy]methyl}imidazol-2-amine (2.00 g, 6.84 mmol) in DMSO (20 mL) was added 2-chloro-4-methylquinazoline (1.22 g, 6.84 mmol) and t-BuOK (1.54 g, 13.7 mmol) at room temperature. The resulting mixture was stirred overnight at 40 °C. The resulting mixture was diluted with water (200 mL). The resulting mixture was extracted with EtOAc (3 x 200 mL). The combined organic layers were washed with brine (3 x 50 mL), dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with PE I EA (30:70) to afford the title compound as a reddish brown solid (900 mg, 30.3%). LCMS: MI 436.Preparation 6 - N-{5-[2-ethoxyethenyl]-1-{[2-(trimethylsilyl)ethoxy]methyl}imidazol-2-yl}-4- methylquinazolin-2-amine

[0183] To a solution of / V-(5-bromo-1-{[2-(trimethylsilyl)ethoxy]methyl}imidazol-2-yl)-4- methylquinazolin-2-amine (900 mg, 2.07 mmol) in 1 ,4-dioxane (9 mL) and H2O (1.8 mL) at room temperature under nitrogen atmosphere was added 2-[(E)-2-ethoxyethenyl]- 4,4,5,5-tetramethyl-1 ,3,2-dioxaborolane (821 mg, 4.14 mmol), K2CO3 (859 mg, 6.21 mmol) and Pd(PPIT)4 (239 mg, 0.207 mmol). The resulting mixture was stirred at 100 °C for additional 6 h. The resulting mixture was concentrated under reduced pressure. The residue was purified by RP-HPLCAto afford the title compound as a reddish brown solid (700 mg, 79.4%). LCMS: MH+426.Preparation 7 - 2-{2-[(4-methylquinazolin-2-yl)amino]-3-{[2-(trimethylsilyl)ethoxy]methyl}imidazol-4-yl}acetaldehyde

[0184] To a solution of of A / -{5-[2-ethoxyethenyl]-1-{[2- (trimethylsilyl)ethoxy]methyl}imidazol-2-yl}-4-methylquinazolin-2-amine (700 mg, 1.65 mmol) in THF (35 mL) was added cone. HCI (12.0 mL). The mixture was stirred at room temperature for 1 h. The mixture was neutralized with sat. NaHCCh (aq.). The mixture was extracted with EtOAc (3 x 50 mL). The combined organic phases were washed with brine, and dried with Na2SO4. After filtration, the filtrate was concentrated under reduced pressure to afford the title compound as a brown oil (700 mg, crude) which was used directly in the following step.Preparation 8 - N-{5-[2-(dimethylamino)ethyl]- 1-{[2-(trimethylsilyl)ethoxy]methyl}imidazol-2- yl}-4-methylquinazolin-2-amine

[0185] To a solution of 2-{2-[(4-methylquinazolin-2-yl)amino]-3-{[2- (trimethylsilyl)ethoxy]methyl}imidazol-4-yl}acetaldehyde (700 mg, 1.76 mmol) in DCM (21 mL) were added dimethylamine hydrochloride (144 mg, 1.76 mmol) and sodium triacetoborohydride (560 mg, 2.64 mmol). The resulting mixture was stirred at room temperature for additional 2 h. The resulting mixture was diluted with water (50 mL). The aqueous layer was extracted with EtOAc (3 x 20 mL). The resulting aqueous layer was concentrated under reduced pressure. The residue was purified by RP-HPLCBto afford the title compound as a light yellow solid (70.0 mg, 9.3%). LCMS: MIT 427.Preparation 9 - N-(5-(2-(dimethylamino)ethyl)-1H-imidazol-2-yl)-4-methylquinazolin-2- amine

[0186] To a solution of N-{5-[2-(dimethylamino)ethyl]-1-{[2- (trimethylsilyl)ethoxy]methyl}imidazol-2-yl}-4-methylquinazolin-2-amine (70.0 mg, 0.164 mmol, 1.00 eq) in THF (2.1 mL) at room temperature was added HCI (1.4 mL, 6.0 M / L). The resulting mixture was stirred at 60 °C for additional 6 h. The mixture was allowed to cool down to room temperature. The crude product was purified by Prep-HPLCDto afford A / -(5- (2-(dimethylamino)ethyl)-1 / 7-imidazol-2-yl)-4-methylquinazolin-2-amine as a light yellow solid (4.5 mg, 9.3%) .1H NMR (300 MHz, DMSO-d6) 5 11.66 (br, 1 H), 10.65 (br, 1 H), 8.09 - 8.06 (m, 1 H), 7.82 - 7.81 (m, 2H), 7.45 - 7.34 (m, 1 H), 6.58 (s, 1 H), 3.34 - 3.22 (m, 2H), 2.82 (s, 3H), 2.63 - 2.52 (m, 2H), 2.21 (s, 6H). LCMSA: Rt 1.38 min, MH+297.Example 2: 4-methyl- / V-(5-(2-morpholinoethyl)-1 / 7-imidazol-2-yl)quinazolin-2-aminePreparation 10 - 2-(2-((4-bromophenyl)diazenyl)-1H-imidazol-5-yl)ethan-1-ol

[0187] To a stirring solution of 4-bromoaniline (5.40 g, 31.4 mmol) in aqueous HCI (160 mL, 2.3 M) at 0 °C was added a solution of NaNC>2 (2.30 g, 33.3 mmol) in water (30 mL) dropwise. The resulting mixture was stirred at 0 °C for 40 min. The above mixture was added into a stirred solution of 2-(1 / 7-imidazol-4-yl)ethanol (5.00 g, 44.6 mmol) in Na2COs (320 mL, 0.2 M) at 0 °C. The resulting mixture was stirred at room temperature for 6 h. The reaction was basified to pH 10 with aq. NaOH (3.0 M). The resulting mixture was extracted with EtOAc (3 x 500 mL). The combined organic layers were dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified bysilica gel column chromatography, eluted with CH2CI2 I MeOH (8:1) to afford the title compound as a reddish solid (3.8 g, crude). LCMS: MIT 297.Preparation 11 - 2-((4-bromophenyl)diazenyl)-5-(2-chloroethyl)-1H-imidazole

[0188] To a stirred solution of 2-(2-((4-bromophenyl)diazenyl)-1 / 7-imidazol-5-yl)ethan-1-ol (2.15 g, 7.29 mmol) in DCM (30.0 mL) at 0 °C was added SOCI2 (8.67 g, 72.9 mmol) dropwise. The resulting mixture was stirred at room temperature for overnight. The reaction was quenched with sat. NaHCCh (aq.) at 0 °C. The resulting mixture was extracted with CH2CI2 (3 x 100 mL). The combined organic layers were washed with brine (2 x 80 mL), dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with PE I EA (5:1) to afford the title compound as a brown solid (1.70 g, 74.4%). LCMS: MH+315.Preparation 12 - 2-((4-bromophenyl)diazenyl)-5-(2-chloroethyl)-1-((2-(trimethylsilyl) ethoxy) methyl) - 1 H -imidazole

[0189] To a stirred mixture of 2-((4-bromophenyl)diazenyl)-5-(2-chloroethyl)-1 / 7-imidazole (750 mg, 2.39 mmol, 1.00 eq) and Et3N (1 .45 g, 14.4 mmol) in THF (20.0 mL) at 0 °C was added [2-(chloromethoxy)ethyl]trimethylsilane (2.39 g, 14.4 mmol). The resulting mixture was stirred at room temperature for 1 h. The reaction was quenched with sat. NaHCOs (aq.) at 0°C. The resulting mixture was extracted with CH2CI2 (3 x 50 mL). The combined organic layers were washed with brine (2 x 40 mL), dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with PE I EA (6:1) to afford the title compound as a brown solid (714 mg, 67.3%). LCMS: MI 445.Preparation 13 - 4-(2-(2-((4-bromophenyl)diazenyl)- 1-((2-(trimethylsilyl)ethoxy) methyl)- 1H- imidazol-5-yl) ethyl) morpholine

[0190] To a mixture of 2-((4-bromophenyl)diazenyl)-5-(2-chloroethyl)-1-((2- (trimethylsilyl)ethoxy)methyl)-1 / 7-imidazole (200 mg, 0.451 mmol) and morpholine (393 mg, 4.51 mmol) in EtOH (4.0 mL) and H2O (4.0 mL) were added KI (150 mg, 0.902 mmol) and CS2CO3 (294 mg, 0.902 mmol). The resulting mixture was stirred at 100 °C for 8 h. Then the reaction was diluted with water (20 mL). The resulting mixture was extracted with CH2CI2 (3 x 30 mL). The combined organic layers were washed with brine (2 x 30 mL), dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with CH2CI21 MeOH (10:1) to afford the title compound as a red oil (135 mg, 60.6%). LCMS: MH+496.Preparation 14 - 5-(2-morpholinoethyl)- 1-((2-(trimethylsilyl)ethoxy)methyl)- 1H-imidazol-2- amine

[0191] To a solution of 4-(2-(2-((4-bromophenyl)diazenyl)-1-((2- (trimethylsilyl)ethoxy)methyl)-1 / 7-imidazol-5-yl)ethyl)morpholine (160 mg, 0.324 mmol) in MeOH (3.0 mL) was added 10% Pd / C (160 mg). The mixture was stirred under hydrogen atmosphere at room temperature for 1 h. The resulting mixture was filtered; the filter cake was washed with MeOH (3 * 10 mL). The combined filtrate was concentrated under reduced pressure to afford the title compound as a yellow oil (113 mg, crude). LCMS: MH+327.Preparation 15 - 4-methyl-N-(5-(2-morpholinoethyl)- 1-((2-(trimethylsilyl)ethoxy) methyl)- 1H- imidazol-2-yl)quinazolin-2-amine

[0192] To a mixture of 5-(2-morpholinoethyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1 / 7- imidazol-2-amine (85.0 mg, 0.260 mmol) in dioxane (1.0 mL) was added Pd(OAc)2 (5.84 mg, 26.0 pmol), Xantphos (15.1 mg, 26.0 pmol), CS2CO3 (170 mg, 0.520 mmol) and 2-chloro-4- methylquinazoline (23.3 mg, 0.130 mmol). The mixture was stirred under nitrogen atmosphere at 80 °C for 4 h. Then the reaction was diluted with water (20 mL). The resulting mixture was extracted with CH2CI2 (3 x 30 mL). The combined organic layers were washed with brine (2 x 20 mL), dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by Prep-TLC, eluted with CH2CI2 / MeOH (15:1) to afford the title compound as a yellow oil (23.0 mg, 18.9%). LCMS: MH+469.Preparation 16 - 4-methyl-N-(5-(2-morpholinoethyl)-1H-imidazol-2-yl)quinazolin-2-amine

[0193] To a stirred solution of 4-methyl- / V-(5-(2-morpholinoethyl)-1-((2- (trimethylsilyl)ethoxy)methyl)-1 / 7-imidazol-2-yl)quinazolin-2-amine (20.0 mg, 43.0 pmol) in DCM (2.0 mL) at room temperature was added TFA (2.0 mL) dropwise. The resulting mixture was stirred at room temperature for 1 h. The resulting mixture was concentrated under reduced pressure. The residue was purified by RP-HPLC0to afford title product as a yellow solid (6.2 mg, 42.9%).1H NMR (300 MHz, DMSO-d6) 5 8.16 (d, J = 8.4 Hz, 1 H), 8.01 - 7.84 (m, 2H), 7.50 (t, J = 7.5 Hz, 1 H), 6.93 (s, 1 H), 3.86 (t, J = 4.7 Hz, 4H), 3.26 - 3.18 (m, 2H), 3.17 - 3.07 (m, 4H), 3.01 - 2.91 (m, 2H), 2.90 (s, 3H). LCMSA: Rt 1.45 min, MH+339.Example 3: A / -(5-(2-(dimethylamino)ethyl)-1 H-imidazol-2-yl)-4-methyl-6,7-dihydro-5H- cyclopenta[cf]pyrimidin-2-aminePreparation 17- 4-methyl-6, 7-dihydro-5H-cyclopenta[d]pyrimidin-2-ol

[0194] To a stirred solution of 2-acetylcyclopentan-1-one (5.00 g, 39.6 mmol) in EtOH (50.0 mL) was added urea (357 mg, 59.5 mmol) and cone. HCI (3.4 mL) at room temperature. The resulting mixture was stirred for 3 h at 80 °C. The precipitated solids were collected by filtration and washed with EtOH (3 x 5 mL). The filter cake was dried under vacuum to afford a brown solid (4.75 g), which was dissolved in aq. NaOH (50.0 mL, 10% wt) and stirred for 3 h at 40 °C. The mixture was neutralized to pH 7 with aq. AcOH. The resulting mixture was extracted with CH2CI2 (3 x 50 mL). The combined organic layers were washed with brine (3 x 10 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with CH2CI2 / MeOH (10:1) to afford the title compound as white solid (2.29 g, 68.7%). LCMS: MH+151.Preparation 18 - 2-chloro-4-methyl-6, 7-dihydro-5H-cyclopenta[d]pyrimidine

[0195] To a solution of 4-methyl-6,7-dihydro-5H-cyclopenta[c(]pyrimidin-2-ol (500 mg, 3.33 mmol) in 1 ,4-dioxane (17.0 mL) was added POCh (0.6 mL, 6.66 mmol) dropwise over 10 min at 0 °C. The resulting mixture was stirred for additional 3 h at 100 °C. The mixture was allowed to cool down to room temperature. The resulting mixture was diluted with water (10 mL) and extracted with CH2CI2 (3 x 20 mL). The combined organic layers were washed with brine (3 x 10 mL), dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with PE I EA (2:1) to afford the title compound as white solid (195 mg, 34.7%). LCMS: MH+169.Preparation 19 - 2-(2-((4-bromophenyl)diazenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H- imidazol-5-yl)-N, N-dimethylethan- 1 -amine

[0196] To a solution of 2-((4-bromophenyl)diazenyl)-5-(2-chloroethyl)-1-((2- (trimethylsilyl)ethoxy)methyl)-1 / 7-imidazole (14.0 g, 31.5 mmol) in EtOH (120.0 mL) and H2O (40.0 mL) was added KI (10.5 g, 63.1 mmol), DIEA (11.0 mL, 63.1 mmol) and dimethylamine hydrochloride (25.7 g, 315 mmol) at 0 °C under nitrogen atmosphere. The resulting mixture was stirred overnight at 80 °C. The mixture was allowed to cool down to room temperature. The resulting mixture was diluted with water (800 mL) and extracted with EtOAc (3 x 800 mL). The combined organic layers were washed with brine (800 mL), dried over anhydrous Na2SC>4. After filtration, the combined filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with CH2CI21 MeOH (10:1) to afford the title compound (6.20 g, 43.4%) as brown oil. LCMS: MH+454.Preparation 20 5-(2-(dimethylamino)ethyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H- imidazol-2-amine

[0197] To a solution of 2-(2-((4-bromophenyl)diazenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)- 1 / 7-imidazol-5-yl)-N,N-dimethylethan-1-amine (6.20 g, 13.7 mmol) in MeOH (90 mL) was added Pd / C (6.20 g, 10% wf) and AcOH (90 pL). The mixture was stirred for 3 h at room temperature under hydrogen atmosphere. The reaction mixture was filtered through a short pad of Celite. The combined filtrate was concentrated under reduced pressure. The residue was purified by RP-HPLCFto afford the title compound (1.70 g, 43.5%) as yellow solid. LCMS: MIT 285.Preparation 21 - N-(5-(2-(dimethylamino)ethyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H- imidazol-2-yl)-4-methyl-6, 7-dihydro-5H-cyclopenta[d]pyrimidin-2-amine

[0198] To a stirred solution of 2-chloro-4-methyl-6,7-dihydro-5H-cyclopenta[c(]pyrimidine (90 mg, 0.53 mmol) and 5-(2-(dimethylamino)ethyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1 / 7- imidazol-2-amine (152 mg, 0.53 mmol) in dioxane (3.00 mL) were added Pd2(dba)s (48.9 mg, 0.05 mmol), Xantphos (30.9 mg, 0.05 mmol) and CS2CO3 (348 mg, 1.07 mmol) at room temperature under nitrogen atmosphere. The resulting mixture was stirred overnight at 90 °C under nitrogen atmosphere. The resulting mixture was filtered, the filter cake was washed with DMF (3 x 5 mL). The combined filtrate was concentrated under reduced pressure. The residue was purified by RP-HPLCEto afford the title compound as brown oil (135 mg, 60.7%). LCMS: MH* 417.Preparation 22 - N-(5-(2-(dimethylamino)ethyl)-1H-imidazol-2-yl)-4-methyl-6, 7-dihydro-5H- cyclopenta[d]pyrimidin-2-amine.

[0199] To a solution of A / -(5-(2-(dimethylamino)ethyl)-1-((2-(trimethylsilyl)ethoxy)methyl)- 1 H-imidazol-2-yl)-4-methyl-6,7-dihydro-5H-cyclopenta[c(]pyrimidin-2-amine (80 mg, 0.19 mmol) in DCM (2.00 mL) was added TFA (2.00 mL) dropwise at 0 °C. The resulting mixture was stirred for 2 h at room temperature. The resulting mixture was concentrated under reduced pressure. The residue was purified by RP-HPLCGto afford the title compound as a white solid (6 mg, 10.9%).1H NMR (400 MHz, DMSO-d6) 5 11.19 (br, 1 H), 10.09 (br, 1 H), 6.47 (s, 1 H), 2.88 - 2.72 (m, 4H), 2.57 (s, 2H), 2.48 - 2.42 (m, 2H), 2.32 (s, 3H), 2.20 (s, 6H), 2.06 - 1 .96 (m, 2H). LCMSB: Rt 0.838 min, MH+287.Example 4: A / -(5-(2-(dimethylamino)ethyl)-1 H-imidazol-2-yl)-4-phenylpyrimidin-2- aminePreparation 23 - N-{5-[2-(dimethylamino)ethyl]- 1-{[2-(trimethylsilyl)ethoxy]methyl}imidazol- 2-yl}-4-phenylpyrimidin-2-amine

[0200] To a solution of 2-chloro-4-phenylpyrimidine (130 mg, 0.682 mmol) and 5-(2- (dimethylamino)ethyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1 / 7-imidazol-2-amine (194 mg, 0.682 mmol) in 1 ,4-dioxane (3.5 mL) were added BINAP (42.5 mg, 0.068 mmol), Pd(OAc)2 (15.3 mg, 0.068 mmol) and CS2CO3 (444 mg, 1.36 mmol) under nitrogen atmosphere at room temperature. The resulting mixture was stirred under nitrogen atmosphere at 90 °C overnight. The mixture was allowed to cool down to room temperature. The resulting mixture was filtered, the filter cake was washed with EtOAc (3 x 5 mL). The combined filtrate was concentrated under reduced pressure. The residue was purified by Prep-HPLCHto afford the title compound as a brown solid (80.0 mg, 26.7%). LCMS: MH+439.Preparation 24 N-(5-(2-(dimethylamino)ethyl)-1H-imidazol-2-yl)-4-phenylpyrimidin-2- amine

[0201] To a solution of / V-{5-[2-(dimethylamino)ethyl]-1-{[2- (trimethylsilyl)ethoxy]methyl}imidazol-2-yl}-4-phenylpyrimidin-2-amine (80.0 mg, 0.182 mmol) in DCM (0.5 mL) was added TFA (0.5 mL) dropwise at room temperature. The resulting mixture was stirred at room temperature for 1 h. The resulting mixture was concentrated under reduced pressure. The residue was purified by Prep-HPLC to afford the title compound as a yellow solid (24.6 mg, 43.7%).1H NMR (400 MHz, DMSO-d6) 5 11.14 (br, 1 H), 10.44 (br, 1 H), 8.55 (d, J = 4.0 Hz, 1 H), 8.18 - 8.10 (m, 2H), 7.59 - 7.51 (m, 3H), 7.44 (d, J = 4.0 Hz, 1 H), 6.56 (s, 1 H), 2.66 - 2.57 (m, 2H), 2.54 (d, J = 4.0 Hz, 2H), 2.22 (s, 6H). LCMSc: Rt 0.77 min, MH+309.Example 5: A / -(5-(2-(dimethylamino)ethyl)-1 H-imidazol-2-yl)-4-methyl-6- phenylpyrimidin-2-aminePreparation 25 - 2-chloro-4-methyl-6-phenylpyrimidine

[0202] To a mixture of 2,4-dichloro-6-methylpyrimidine (1.00 g, 6.13 mmol), phenyl boronic acid (600 mg, 4.90 mmol) in dioxane (12.0 mL) and H2O (4.0 mL) were added Pd(dppf)Ch (450 mg, 0.610 mmol) and CS2CO3 (4.00 g, 12.2 mmol). The mixture was irradiated with microwave radiation under nitrogen atmosphere at 70 °C for 15 min. The mixture was allowed to cool down to room temperature. The resulting mixture was diluted with water (50 mL) and extracted with EtOAc (3 x 50 mL). The combined organic layers were washed with brine (2 x 30 mL), dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with PE / EA (10:1) to afford the title compound (610 mg, 48.5%) as a white solid. LCMS: MIT 205.Preparation 26 - N-{5-[2-(dimethylamino)ethyl]- 1-{[2-(trimethylsilyl)ethoxy]methyl}imidazol- 2-yl}-4-methyl-6-phenylpyrimidin-2-amine

[0203] To a mixture of 5-(2-(dimethylamino)ethyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1 / 7- imidazol-2-amine (100 mg, 0.350 mmol) and 2-chloro-4-methyl-6-phenylpyrimidine (71.9 mg, 0.350 mmol) in dioxane (2.0 mL) were added Pd2(dba)s (32.1 mg, 30.0 pmol), XantPhos (20.3 mg, 30.0 pmol) and CS2CO3 (229 mg, 0.700 mmol). The mixture was stirred under nitrogen atmosphere at 100 °C for 2 h. The mixture was allowed to cool down to room temperature. The resulting mixture was filtered, the filter cake was washed with CH2CI2 (3 x 10 mL). The combined filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with CH2CI21 MeOH (20:1) to afford the title compound as a white solid (105 mg, 65.9%). LCMS: MIT 453.Preparation 27 - N-(5-(2-(dimethylamino)ethyl)-1H-imidazol-2-yl)-4-methyl-6- phenylpyrimidin-2-amine

[0204] A solution of / V-{5-[2-(dimethylamino)ethyl]-1-{[2-(trimethylsilyl)ethoxy]methyl}imidazol-2-yl}-4-methyl-6-phenylpyrimidin-2-amine (100 mg, 0.220 mmol) in TFA (2.0 mL) and DCM (4.0 mL) was stirred at room temperature for 2 h. The resulting mixture was concentrated under reduced pressure. The crude product was purified by RP-HPLCLto afford the title compound as a light-yellow semi-solid trifluoracetic acid salt(23.2 mg, 24.0%).1H NMR (300 MHz, DMSO-d6) 5 8.18 - 8.12 (m, 2H), 7.64 (s, 1 H), 7.60 - 7.53 (m, 3H), 7.11 (s, 1 H), 3.36 (t, J = 9.0 Hz, 2H), 3.04 (t, J = 9.0 Hz, 2H), 2.83 (s, 6H), 2.57 (s, 3H). LCMSE: Rt 1.05 min, MH+323.Example 6: 2-((5-(2-(dimethylamino)ethyl)-1 / 7-imidazol-2-yl)amino)-4,6- dimethylnicotinonitrilePreparation 28 - 2-((5-(2-(dimethylamino)ethyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H- imidazol-2-yl)amino)-4,6-dimethylnicotinonitrile

[0205] To a mixture of 5-(2-(dimethylamino)ethyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1 / 7- imidazol-2-amine (150 mg, 0.527 mmol) and 2-chloro-4,6-dimethylnicotinonitrile (87.9 mg, 0.527 mmol) in toluene (3.0 mL) were added Pd2(dba)s (48.3 mg, 53.0 pmol), XantPhos (30.5 mg, 53.0 pmol) and CS2CO3 (344 mg, 1.05 mmol). The mixture was stirred under nitrogen atmosphere at 100 °C overnight. The mixture was allowed to cool down to room temperature and quenched with water. The resulting mixture was extracted with CH2CI2 (3 x 30 mL). The combined organic layers were washed with brine (20 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluted with CH2CI2 1 MeOH (10:1)) to afford the title compound as a yellow oil (94.0 mg, 43.0%). LCMS: MH+415.Preparation 29 - 2-((5-(2-(dimethylamino)ethyl)-1H-imidazol-2-yl)amino)-4,6- dimethylnicotinonitrile

[0206] To a solution of 2-((5-(2-(dimethylamino)ethyl)-1-((2-(trimethylsilyl)ethoxy)methyl)- 1 / 7-imidazol-2-yl)amino)-4,6-dimethylnicotinonitrile (65.0 mg, 0.157 mmol) in DCM (2.0 mL) was added TFA (1.0 mL) dropwise at room temperature. The resulting mixture was stirred at room temperature for 3 h and concentrated under reduced pressure. The resulting mixture was purified by Prep-HPLCMto afford the title compound as a white solid (4.8 mg, 10.8%).1H NMR (400 MHz, Methanol-d4) 5 7.50 (s, 1 H), 6.90 (s, 1 H), 2.88 - 2.82 (m, 5H), 2.74 (t, J = 7.6 Hz, 2H), 2.50 (s, 3H), 2.34 (s, 6H). LCMSF: Rt 0.912 min, MH+285.Example 7: A / -(5-(3-aminopropyl)-1H-imidazol-2-yl)-4-methylquinazolin-2-aminePreparation 30 - tert-butyl N-[3-{2-[(4-methylquinazolin-2-yl)amino]-3-{[2-(trimethylsilyl)ethoxy]methyl}imidazol-4-yl}prop-2-en-1-yl]carbamate

[0207] To a mixture of / V-(5-bromo-1-{[2-(trimethylsilyl)ethoxy]methyl}imidazol-2-yl)-4- methylquinazolin-2-amine (490 mg, 1.12 mmol) in 1 ,4-dioxane / H2O (6.00 mL, 5 / 1) was added terf-butyl / V-[3-(4,4,5,5-tetramethyl-1 ,3,2-dioxaborolan-2-yl)prop-2-en-1-yl]carbamate (1.28 g, 4.51 mmol), XantPhos Pd G2 (100 mg, 0.11 mmol), XPhos (53.8 mg, 0.11 mmol) and K2CO3 (312 mg, 2.25 mmol). The mixture was stirred under nitrogen atmosphere at 120 °C for 3 h. The resulting mixture was diluted with water (20 mL). The resulting mixture was extracted with EtOAc (3 x 20 mL). The combined organic layers were washed with brine (2 x 100 mL), dried over anhydrous Na2SO4. After filtration, the combined filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with CH2CI2 1 MeOH (10:1) to afford the title compound as a brown oil (645 mg, crude). LCMS: MIT 511.Preparation 31 - tert-butyl (3-(2-((4-methylquinazolin-2-yl)amino)-1-((2-(trimethylsilyl) ethoxy) methyl) - 1 H-imidazol-5-yl) propyl) carbamate

[0208] To a mixture of terf-butyl / V-[3-{2-[(4-methylquinazolin-2-yl)amino]-3-{[2- (trimethylsilyl)ethoxy]methyl}imidazol-4-yl}prop-2-en-1-yl]carbamate (640 mg, 1.25 mmol) in MeOH (50.0 mL) and AcOH (5.0 mL) was added Raney-Ni (814 mg). The mixture was stirred at room temperature under hydrogen atmosphere overnight. The resulting mixture was filtered through a pad of Celite. The combined filtrate was concentrated under reduced pressure. The residue was diluted with DCM (40 mL) and sat. NaHCOs(40 mL). The resulting mixture was extracted with DCM (30 mL x3). The combined organic layers were dried over Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. This resulted in the title compound as a brown oil (644 mg, crude). LCMS: MH+513.Preparation 32 - N-(5-(3-aminopropyl)-1H-imidazol-2-yl)-4-methylquinazolin-2-amine

[0209] To a solution of terf-butyl (3-(2-((4-methylquinazolin-2-yl)amino)-1-((2- (trimethylsilyl)ethoxy)methyl)-1 H-imidazol-5-yl)propyl)carbamate (100 mg, 0.19 mmol) in DCM (4.0 mL) was added TFA (2.0 mL). The reaction mixture was stirred at room temperature for 2 h. The resulting mixture was concentrated under reduced pressure. The crude product (90 mg) was purified by RP-HPLCNto afford the title compound as a whitesolid trifluoracetic acid salt (8.6 mg, 11.12%).1H N MR (400 MHz, Methanol-d4) 5 8.22 - 8.15 (m, 1 H), 7.93 - 7.89 (m, 2H), 7.56 (t, 1 H), 6.96 (s, 1 H), 3.07 - 3.00 (m, 2H), 2.96 (s, 3H), 2.81 (t, J = 7.7 Hz, 2H), 2.12 - 2.00 (m, 2H). LCMSG: Rt 0.75 min, MH+283.Example 8: Methyl 2-((5-(2-(dimethylamino)ethyl)-1 / 7-imidazol-2-yl)amino)pyrimidine- 5-carboxylatePreparation 33 - Methyl 2-((5-(2-(dimethylamino)ethyl)-1-((2-(trimethylsilyl)ethoxy)methyl)- 1H-imidazol-2-yl)amino)pyrimidine-5-carboxylate

[0210] To a solution of methyl 2-chloropyrimidine-5-carboxylate (91.0 mg, 0.527 mmol) and 5-(2-(dimethylamino)ethyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1 / 7-imidazol-2-amine (150 mg, 0.527 mmol) in toluene (5.0 mL) at room temperature under nitrogen atmosphere were added Pd2(dba)s (48.3 mg, 0.053 mmol), XantPhos (30.5 mg, 0.053 mmol) and CS2CO3 (344 mg, 1.05 mmol). The mixture was stirred at 100 °C overnight. The reaction was quenched with water at room temperature. The resulting mixture was extracted with CH2CI2 (5 x 20 mL). The combined organic layers were washed with brine (3 x 20 mL), dried over anhydrous Na2SC>4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by Prep-TLC (CH2CI2 / MeOH=10:1) to afford the title compound as a yellow oil (60 mg, crude). LCMS: MH+421.Preparation 34 - Methyl 2-((5-(2-(dimethylamino)ethyl)-1H-imidazol-2-yl)amino)pyrimidine- 5-carboxylate

[0211] A solution of methyl 2-((5-(2-(dimethylamino)ethyl)-1-((2- (trimethylsilyl)ethoxy)methyl)-1 H-imidazol-2-yl)amino)pyrimidine-5-carboxylate (50.0 mg, 0.119 mmol) in TFA (0.5 mL) and DCM (1.0 mL) was stirred at room temperature for 2 h. The resulting mixture was concentrated under reduced pressure. The mixture was neutralized to pH 7 with NH3 H2O. The crude product was purified by RP-HPLCMto afford the title compound as a yellow solid (3.4 mg, 8.8%).1H NMR (400 MHz, Methanol-d4) 5 8.99 (s, 2H), 6.65 (s, 1 H), 3.91 (s, 3H), 2.74 - 2.70 (m, 2H), 2.70 - 2.57 (m, 2H), 2.33 (s, 6H). LCMSH: Rt 0.744 min, MH+291.Example 9: A / -(5-(2-aminoethyl)-1 / 7-imidazol-2-yl)-4-methylquinazolin-2-aminePreparation 35 - 2-(2-(2-((4-methylquinazolin-2-yl)amino)-1H-imidazol-5- yl)ethyl)isoindoline- 1, 3-dione

[0212] To a stirred mixture of A / -(4-methylquinazolin-2-yl)guanidine (200 mg, 0.994 mmol) and 2-(4-bromo-3-oxobutyl)isoindole-1 , 3-dione (294 mg, 0.994 mmol) in DMSO (4.00 mL) was added K2CO3 (274 mg, 1.98 mmol) at room temperature. The resulting mixture was stirred at room temperature for 1 h. The reaction was quenched by the addition of water (30 mL). The reaction mixture was filtered through a short pad of Celite. The pad was washed with water (10 mL x 3). The combined filtrate was concentrated under reduce pressure to afford the title compound as a brown solid (320 mg, crude). LCMS: MIT 399.Preparation 36 - N-(5-(2-aminoethyl)-1H-imidazol-2-yl)-4-methylquinazolin-2-amineTo a mixture of 2-(2-(2-((4-methylquinazolin-2-yl)amino)-1 / 7-imidazol-5-yl)ethyl)isoindoline- 1 , 3-dione (260 mg, 0.653 mmol) in EtOH (5.2 mL) were added hydrazine hydrate (866 uL, 17.8 mmol) at room temperature. The resulting mixture was stirred at 80 °C for 1 h. The resulting mixture was concentrated under reduced pressure. The residue was purified by RP- HPLCNto afford the title compound as a white solid (9.7 mg, 4.86%).1H NMR (300 MHz, DMSO-d6) 5 8.27 (d, J = 8.3 Hz, 1 H), 8.07 - 7.88 (m, 2H), 7.73 - 7.58 (m, 1 H), 7.30 (s, 1 H), 7.15 (s, 2H), 3.25 - 3.13 (m, 1 H), 2.97 (s, 3H), 2.85 - 2.70 (m, 1 H), 2.48 - 2.39 (m, 2H). LCMS1: Rt 0.56 min, MH+269.Example 10: A / -(5-(2-(dimethylamino)ethyl)-1H-imidazol-2-yl)pyrimidin-2-aminePreparation 37 - N-(5-(2-(dimethylamino)ethyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H- imidazol-2-yl)pyrimidin-2-amine

[0213] To a mixture of 5-(2-(dimethylamino)ethyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1 / 7- imidazol-2-amine (150 mg, 0.528 mmol) and 2-chloropyrimidin (60 mg, 0.528 mmol) in toluene (3.0 mL) at room temperature were added Pd2(dba)s (48 mg, 0.053 mmol), XantPhos (30 mg, 0.053 mmol) and CS2CO3 (344 mg, 1.05 mmol). The resulting mixture was stirred under nitrogen atmosphere at 100 °C overnight. The mixture was allowed to cool down toroom temperature. The resulting mixture was diluted with water (120 mL) and extracted with CH2CI2 (3 x 60 mL). The combined organic layers were washed with brine (2 x 10 mL), dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by Prep-TLC (CH2CI2 I MeOH 8:1) to afford the title compound as a brown oil (57 mg, 29.8%). LCMS: MIT 363.Preparation 38 - N-(5-(2-(dimethylamino)ethyl)-1H-imidazol-2-yl)pyrimidin-2-amine

[0214] To a stirred solution of A / -(5-(2-(dimethylamino)ethyl)-1-((2- (trimethylsilyl)ethoxy)methyl)-1 / 7-imidazol-2-yl)pyrimidin-2-amine (55 mg, 0.15 mmol) in DCM (1.5 mL) at room temperature was added TFA (1.0 mL). The resulting mixture was stirred at room temperature for 1 h. The resulting mixture was concentrated under reduced pressure. The residue was purified by RP-HPLC° to afford the title compound (24.7 mg, 70.1%).1H NMR (400 MHz, Methanol-d4) 5 8.50 (d, J = 4.8 Hz, 2H), 6.90 (t, J = 4.8 Hz, 1 H), 6.62 (s, 1 H), 2.82 - 2.7 (m, 4H), 2.41 (s, 6H). LCMSJ: Rt 1.313 min, MH+233.Example 11 : A / -(5-(2-(dimethylamino)ethyl)-1 H-imidazol-2-yl)-4-methylpyrimidin-2- aminePreparation 39 - N-(5-(2-(dimethylamino)ethyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H- imidazol-2-yl)-4-methylpyrimidin-2-amine

[0215] To a mixture of 5-(2-(dimethylamino)ethyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1 / 7- imidazol-2-amine (100 mg, 0.352 mmol) and 2-chloro-4-methylpyrimidine (45.1 mg, 0.352 mmol) in toluene (2.0 mL) were added Pd2(dba)s (32.2 mg, 35.2 pmol), XantPhos (20.4 mg, 35.2 pmol) and CS2CO3 (114 mg, 0.352 mmol) at room temperature. The mixture was stirred under nitrogen atmosphere at 100 °C overnight. The mixture was allowed to cool down to room temperature. The reaction was quenched by the addition of water (20 mL) at room temperature. The resulting mixture was extracted with CH2CI2 (3 x 20 mL). The combined organic layers were washed with brine (20 mL), dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by Prep-TLC, eluted with DCM / MeOH (10 / 1), to afford the title compound as a yellow oil (35.0 mg, 28.6%). LCMS: MH+377.Preparation 40 N-(5-(2-(dimethylamino)ethyl)-1H-imidazol-2-yl)-4-methylpyrimidin-2- amine

[0216] To a stirred solution of A / -(5-(2-(dimethylamino)ethyl)-1-((2- (trimethylsilyl)ethoxy)methyl)-1 / 7-imidazol-2-yl)-4-methylpyrimidin-2-amine (37 mg, 98.0 pmol) in DCM (2.0 mL) was added TFA (1.0 mL) at room temperature. The resulting mixture was stirred for 2 h at room temperature. The resulting mixture was concentrated under reduced pressure. The residue was neutralized to pH 7 with NH3.H2O. The residue was purified by RP-HPLCKto afford the title compound as a white solid (6.6 mg, 27.3%).1H NMR (400 MHz, Methanol-d4) 6 8.34 (d, J = 5.1 Hz, 1 H), 6.80 (d, J = 5.1 Hz, 1 H), 6.60 (s, 1 H), 2.78 - 2.62 (m, 4H), 2.45 (s, 3H), 2.34 (s, 6H). LCMSK: Rt 0.90 min, MH+247.Example 12: 4-(2-(2-((4-methylquinazolin-2-yl)amino)-1 H-imidazol-5- yl)ethyl)thiomorpholine 1,1 -dioxidePreparation 41 - 4-(2-(2-((4-bromophenyl)diazenyl)- 1-((2-(trimethylsilyl)ethoxy) methyl)- 1H- imidazol-5-yl)ethyl)thiomorpholine 1, 1 -dioxide

[0217] To a stirred mixture of 2-((4-bromophenyl)diazenyl)-5-(2-chloroethyl)-1-((2- (trimethylsilyl)ethoxy)methyl)-1 / 7-imidazole (300 mg, 0.679 mmol) and thiomorpholine 1 ,1- dioxide (1.83 g, 13.5 mmol) in EtOH (6.0 mL) and H2O (2.0 mL) at room temperature were added KI (224 mg, 1.35 mmol) and DIEA (175 mg, 1.35 mmol). The resulting mixture was stirred at 80 °C for 16 h. The mixture was allowed to cool down to room temperature. The resulting mixture was concentrated under reduced pressure. The resulting mixture was diluted with water (200 mL) and extracted with EtOAc (3 x 100 mL). The combined organic layers were washed with brine (2 x 20 mL), dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with PE I EA (1 :1) to afford the title compound (320 mg, 87.1%) as orange oil. LCMS: MH+544.Preparation 42 - 4-(2-(2-amino-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-imidazol-5- yl)ethyl)thiomorpholine 1, 1 -dioxide

[0218] A mixture of 4-(2-(2-((4-bromophenyl)diazenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)- 1 H-imidazol-5-yl)ethyl)thiomorpholine 1 ,1 -dioxide (308 mg, 0.569 mmol) and Pd / C (308 mg, 10% wt) in MeOH (9.0 mL) was stirred under hydrogen atmosphere at room temperature for 3 h. The resulting mixture was filtered, the filter cake was washed with MeOH (3 x 5 mL). The filtrate was concentrated under reduced pressure. This resulted in the title compound (235 mg, crude) as orange oil. LCMS: MH+375.Preparation 43 - 4-(2-(2-((4-methylquinazolin-2-yl)amino)-1-((2-(trimethylsilyl)ethoxy)methyl)- 1H-imidazol-5-yl)ethyl)thiomorpholine 1, 1 -dioxide

[0219] To a stirred mixture of 4-(2-(2-amino-1-((2-(trimethylsilyl)ethoxy)methyl)-1 / 7- imidazol-5-yl)ethyl)thiomorpholine 1 ,1 -dioxide (230 mg, 0.615 mmol) and 2-chloro-4- methylquinazoline (110 mg, 0.615 mmol) in 1 ,4-dioxane (5.0 mL) at room temperature were added Pd(OAc)2 (13.8 mg, 0.061 mmol), Xantphos (35.5 mg, 0.061 mmol) and CS2CO3 (400 mg, 1.23 mmol). The resulting mixture was stirred under nitrogen atmosphere at 100 °C for 2 h. The mixture was allowed to cool down to room temperature. The resulting mixture was diluted with water (150 mL) and extracted with EtOAc (3 x 60 mL). The combined organic layers were washed with brine (2 x 20 mL), dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by Prep-TLC (CH2CI2 1 MeOH 30:1) to afford the title compound (96 mg, 33.4%, over two steps) as an orange semi-solid. LCMS: MIT 517.Preparation 44 - 4-(2-(2-((4-methylquinazolin-2-yl)amino)-1H-imidazol-5- yl)ethyl)thiomorpholine 1, 1 -dioxide

[0220] To a stirred solution of 4-(2-(2-((4-methylquinazolin-2-yl)amino)-1-((2- (trimethylsilyl)ethoxy)methyl)-1 / 7-imidazol-5-yl)ethyl)thiomorpholine 1 ,1 -dioxide (95 mg, 0.184 mmol) in DCM (2.0 mL) at room temperature was added TFA (2.0 mL) dropwise. The resulting mixture was stirred at room temperature for 1 h. The resulting mixture was concentrated under reduced pressure. The resulting mixture was diluted with water (5 mL) and DCM (5 mL). The mixture was basified to pH 8 with saturated NaHCOs (aq.). The resulting mixture was diluted with water (30 mL) and extracted with CH2CI2 (3 x 40 mL). The combined organic layers were washed with brine (2 x 10 mL), dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by RP-HPLC0to afford the title compound (57.7 mg, 81.3%) as a yellow solid.1H NMR (400 MHz, DMSO-d6) 5 11.6 (br, 1 H), 10.6 (br, 1 H), 8.16 - 7.99 (m, 1 H), 7.92 - 7.73 (m, 2H), 7.47 - 7.30 (m, 1 H), 6.66 (s, 1 H), 3.16 - 3.05 (m, 4H), 3.03 - 2.90 (m, 4H), 2.81 (s, 3H), 2.79 - 2.72 (m, 2H), 2.69 - 2.56 (m, 2H). LCMSL: Rt 1.52 min, MH+387.Example 13: A / , / V-dimethyl-3-(2-((4-methylquinazolin-2-yl)amino)-1H-imidazol-5- yl)propanamidePreparation 45 - ethyl 3-(2-((4-methylquinazolin-2-yl)amino)-1-((2-(trimethylsilyl) ethoxy) methyl) - 1 H-imidazol-5-yl) acrylate

[0221] To a mixture of / V-(5-bromo-1-{[2-(trimethylsilyl)ethoxy]methyl}imidazol-2-yl)-4- methylquinazolin-2-amine (1.20 g, 2.76 mmol), ethyl-3-(4,4,5,5-tetramethyl-1 ,3,2- dioxaborolan-2-yl)prop-2-enoate (2.47 mL, 11.05 mmol), XantPhos Pd G2 (245 mg, 0.280 mmol), XPhos (132 mg, 0.280 mmol) and K2COs (764 mg, 5.52 mmol) in 1 ,4-dioxane / H2O (5:1) (12.0 mL). The mixture was stirred under nitrogen atmosphere at 120 °C overnight. The resulting mixture was diluted with water (100 mL). The resulting mixture was extracted with EtOAc (3 x 100 mL). The combined organic layers were washed with brine (3 x 200 mL), dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with PE I EA (2:1) to afford the title compound as orange oil (667 mg, 51.1%). LCMS: MIT 454.Preparation 46 - ethyl 3-(2-((4-methylquinazolin-2-yl)amino)-1-((2-(trimethylsilyl) ethoxy) methyl) - 1 H-imidazol-5-yl) pro pa noate

[0222] To a mixture of ethyl 3-(2-((4-methylquinazolin-2-yl)amino)-1-((2- (trimethylsilyl)ethoxy)methyl)-1 / 7-imidazol-5-yl)acrylate (620 mg, 1.36 mmol) and Raney-Ni (1.12 g) in MeOH (50.0 mL) was added AcOH (1.00 mL). The mixture was stirred under hydrogen atmosphere at room temperature overnight. The resulting mixture was filtered, the filter cake was washed with MeOH (3 x 5 mL). The combined filtrate was concentrated under reduced pressure to afford the title compound as orange oil (680 mg, crude). The crude product was used in the next step directly without further purification. LCMS: MH+456.Preparation 47 - 3-(2-((4-methylquinazolin-2-yl)amino)- 1-((2-(trimethylsilyl)ethoxy)methyl)- 1 H-imidazol-5-yl) propanoic acid

[0223] To a solution of ethyl 3-(2-((4-methylquinazolin-2-yl)amino)-1-((2- (trimethylsilyl)ethoxy)methyl)-1 / 7-imidazol-5-yl)propanoate (670 mg, 1.47 mmol) in H2O / THF (1 :1) (40.0 mL) was added UOH.H2O (617 mg, 14.7 mmol). The mixture was stirred at 60 °C for 2 h. The resulting mixture was removed THF under reduced pressure. The mixture was neutralized to pH=7 with aq. HCI (1.0 M) at 0 °C, and extracted with EtOAc (5 x 100 mL). The combined organic layers were dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure to afford the title compound as orange oil (532 mg, crude). The crude product was used in the next step directly without further purification. LCMS: MH+428.Preparation 48 - N,N-dimethyl-3-(2-((4-methylquinazolin-2-yl)amino)-1-((2-(trimethylsilyl) ethoxy) methyl) - 1 H-imidazol-5-yl) propanamide

[0224] To a solution of 3-(2-((4-methylquinazolin-2-yl)amino)-1-((2- (trimethylsilyl)ethoxy)methyl)-1 / 7-imidazol-5-yl)propanoic acid (500 mg, 1.17 mmol) and dimethylamine hydrochloride (191 mg, 2.33 mmol) in DMF (20 mL) were added HATLI (667 mg, 1.75 mmol) and DIEA (0.61 mL, 3.51 mmol). The mixture was stirred at room temperature for 2 h. The resulting mixture was diluted with water (50 mL) and extracted with EtOAc (3 x 50 mL). The combined organic layers were washed with brine (3 x 100 mL), dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by Prep-HPLCpto afford the title compound as orange oil (248 mg, 46.6%). LCMS: MIT 455.Preparation 49 - N,N-dimethyl-3-(2-((4-methylquinazolin-2-yl)amino)-1H-imidazol-5- yl)propanamide

[0225] To a solution of N,N-dimethyl-3-(2-((4-methylquinazolin-2-yl)amino)-1-((2- (trimethylsilyl)ethoxy)methyl)-1 H-imidazol-5-yl)propanamide (50.0 mg, 0.11 mmol) in DCM (4.0 mL) was added TFA (2.0 mL). The mixture was stirred at room temperature for 2 h. The resulting mixture was concentrated under reduced pressure and diluted with water (20 mL), then neutralized to pH=7 with saturated solution of NH4HCO3. The aqueous layer was extracted with CH2CI2 (3 x 20 mL). The combined organic layers were dried over anhydrous Na2SC>4. After filtration, the filtrate was concentrated under reduced pressure. The crude product was purified by Prep-HPLCQto afford the title compound as a light yellow solid (11.0 mg, 30.4%).1H NMR (400 MHz, Methanol-d4) 5 8.06 (d, J = 8.2 Hz, 1 H), 7.93 (d, J = 8.4 Hz, 1 H), 7.83 - 7.75 (m, 1 H), 7.46 - 7.37 (m, 1 H), 6.59 (s, 1 H), 3.04 (s, 3H), 2.97 (s, 3H), 2.92 - 2.85 (m, 5H), 2.73 (t, J = 7.0 Hz, 2H). LCMSM: Rt 1.34 min, MH+325.Biological testingLuciferase read-through assay

[0226] The read-through activity of compounds was assessed using the ADXC8 luciferase reporter cell line (generated as described in McElroy et al. PLoS Biology (2013), 11 , e1001593). Active compounds promote translational read-through of the R223X stop codon in the mutated luciferase gene which leads to generation of full-length luciferase in the compound-treated cells. The amount of luciferase is measured by adding a lysis buffer containing luciferin substrate and detecting luminescence using the following protocol:

[0227] ADXC8 cells in assay media (DMEM+10% FBS) were seeded into 96-well tissue culture treated white plates (Greiner) to a density of 13,000 cells per well and incubated for 24 hours at 37 °C in an atmosphere of 5% CO2. Compounds were solubilised in DMSO at a top concentration of 10 mM, serially diluted with assay media to 25 times final concentrationand added to the plate to achieve a range of final concentrations of 100 pM to 0.045 pM in a final volume of 100 pL. Plates were incubated for a further 24 hours at 37 °C in an atmosphere of 5% CO2. Cells treated with 200 pM G418 (with 1 % DMSO) were used as a positive control.

[0228] Cell viability and firefly luciferase activity were measured in the same assay wells. CellTiter-Fluor™ reagent was prepared as per manufacturer’s instructions (Promega E7120) and 20 pl of 5X CellTiter-Fluor™ reagent was added to all wells, briefly mixed by orbital shaking (300-500 rpm for ~30 seconds) and the plates incubated for 30-45 minutes at 37 °C in an atmosphere of 5% CO2. Cell viability was measured by monitoring fluorescence on a CLARIOstar® plate reader (BMG Labtech). Cells containing 1% DMSO were used as a positive control and cells with 400 pM Doxorubicine were used as negative control to define 100 and 0% respectively. ONE-Glo™ luciferase assay buffer (Promega E7120) was prepared according to manufacturer’s instructions and 100 pl of luciferase reagent was added to each well and the plate incubated at RT for 3 mins before measuring the luminescence on the CLARIOstar® plate reader (BMG Labtech). Cells treated with 200 pM G418 (with 1 % DMSO) were used as a positive control and arbitrarily assigned a value of 100%. Cells treated with 1% DMSO were used as the negative control to define 0% response.

[0229] Concentration effect curves were analysed using non-linear regression using the CDD Vault from Collaborative Drug Discovery (Burlingame, CA). Where datapoints at the highest concentrations tested showed bell-shaped dose-response behaviour (due to cell toxicity confirmed by viability assay), they were removed from the analysis. The bottom and top of the curves were constrained to 0 and the maximum measured response respectively. From the fitted curves, the software derived C100% values (the concentration giving the same response as 200 pM G418) for each compound. The negative logarithm of C100% is reported as pC100% (Table 1). In addition to pC100% values, the maximum fold response relative to 200 pM G418 (Fold over Control) was also captured (Table 1). A pC100% categorisation ofin Table 1 represents examples in which a pC100% response was not reached when assayed at a concentration of 100 pM.Table 1 - Luciferase Assay Data for Selected ExamplesKey- pC100% < 4.0 - Fold over Ctrl > 0 < 0.2+ pC100% > 4.0 < 4.5 # Fold over Ctrl > 0.2 < 1++ pC100%F > 4.5 < 5.0 ## Fold over Ctrl >1 < 5+++ pC100% > 5.0 < 5.5 ### Fold over Ctrl >5 < 10++++ pC100% > 5.5 < 6.0 #### Fold over Ctrl >10 < 15+++++ pC100% > 6.0 ##### Fold over Ctrl >15Read-through of R213X mutation in HDQ-P1 cells by Western Blot

[0230] The ability of compounds to promote read-through in cells was assessed using the HDQ-P1 cell line which contain the R213X mutation in the p53 gene. Active compounds promote translational read-through of the premature stop codon encoded by this mutation, resulting in the detection of full length p53 protein by Western blot. An increase in truncated p53 may also be observed as a result of compounds suppressing nonsense mediated decay of mRNA.

[0231] HDQ-P1 cells (R213X) were treated with DMSO (0.3%), or test article for 24 h at various concentrations. Cells were washed once with ice cold 1X PBS and lysed in MRC lysis buffer (1 M Tris-HCI, pH 7.4, 0.5 M EDTA, 0.25 mM EGTA (Ethylene glycol-bis(2- aminoethylether)-N,N,N',N'-tetraacetic acid), 0.5 M sodium fluoride, 0.1 M sodium pyrophosphate, 0.2 M sodium 2-glycerol 1 -phosphate, 0.2 M sodium orthovanadate, 0.27 M sucrose, 10% (v / v) Triton X-100, 0.1 mM phenylmethylsulphonyl fluoride, 1 M dithiothreitol) and protease inhibitor mixture (Roche) centrifuged at 4 °C for 15 min at 16000xg, the supernatants, termed cell extracts, were collected and total proteins were quantified with Bradford protein assay kit (ThermoFisher). Cell extracts were denatured by incubation in NuPAGE LDS sample buffer for 10 min at 95 °C. 25 pg total protein from HDQ-1 cells were subjected to sodium dodecyl sulfatepolyacrylamide-gel electrophoresis in 4-12% pre-castBis-Tris gels (ThermoFisher). Proteins were transferred onto PVDF membranes, and membranes were blocked for 1 hr at room temperature in 5% skimmed milk powder in Trisbuffered saline with 0.1 % Tween® 20 Detergent (TBST) and incubated overnight with the primary monoclonal antibody DO-1 (sc-126, Santa Cruz Biotechnology, 1 / 1000). After three washes in TBST, the membranes were incubated with the secondary antibody (horseradish peroxidase-conjugated anti-mouse IgG [1 / 10,000]) for 1hr at room temperature. The membranes were washed three times with TBST and chemiluminescence was detected with ECL Western Blot reagent kit (Cytiva-VWR) or Clarity Max ECL substrate (Bio-rad). The signal was quantified with Imaged software (NIH).Figure 1 shows the effect of treatment of HDQ-P1 cells with Example 1 (1.23 pM, 3.7 pM, 11 pM, or 33 pM) compared to DMSO (0.3%), and G418 (200 pM).PAMPA Permeability

[0232] The ability of compounds to cross artificial membrane barriers was assessed using a lipid-PAMPA assay.

[0233] 6 pL of a 1.8% lecithin / dodecane mixture was pipetted into the top compartment of each PTFE acceptor plate (EMD Millipore) well (top compartment), avoiding pipette tip contact with the membrane. 300 pL of PBS (pH 7.4) solution was then added within 10 minutes to each well of the acceptor plate. 300 pL of test compound containing solutions (diluted from 10 mM DMSO stock to final concentration of 10 pM with PBS (pH7.4)) to each well of the donor filter plate (bottom compartment, EMD Millipore). The acceptor plate was carefully placed into the donor plate, making sure the underside of the membrane is in contact with the test compound containing solutions in all wells. The plates are incubated with lids at 25 °C, 60 rpm for 16 hours. After incubation, aliquots of 50 pL from each well of acceptor and donor plate are transferred into a 96-well plate. 200 pL of methanol (containing internal standards: 100 nM Alprazolam, 200 nM caffeine and 100 nM Tolbutamide) was added to each well. Plates were vortexed with lids at 750 rpm for 100 seconds. Samples were centrifuged at 3,800 g for 20 minutes. Compound concentration was assessed by LC / MS / MS analysis by comparison with matrix matched standard curve. The effective permeability (Pe), in units of centimeter per second, can be calculated using the following equation and are then converted and reported for Examples as nanometers per second (nm / s) (Table 2):Where [Drug]aCceptor is the concentration for compounds in the acceptor plate; [Drug]equiiibrium is the theoretical equilibrium concentration after incubation and is given by the formula:([Drug]DonorxVD+ [Drug]acceptorxVA) / (VD+ VA)Where [Drugjoonor is the concentration for compounds in the donor plate; VD is the volume of donor compartment (0.30 mL) and VA is the volume of acceptor compartment (0.30 mL).And C is a constant given by the formula:VDXVA / [(VD+ VA)xtxA]Where A is the filter area (0.24 cm2for Multi-Screen Permeability Filter plate) and t is the incubation time (in seconds).Table 2 - PAMPA Permeability for Selected ExamplesKey+ Pe= 0 < 10 nm / s++ Pe> 10 < 50 nm / s+++ Pe> 50 < 100 nm / s++++ Pe> 100 nm / sThus, all compounds tested showed good permeability.CaCo-2 Permeability

[0234] The ability of compounds to cross cellular membrane barriers was assessed using a bi-directional CaCo-2 assay.

[0235] 50 pL and 25 mL of cell culture medium (Dulbecco’s Modified Eagle’s Medium (DMEM, Corning Corporation) with high glucose and L-glutamine supplemented with: 10% FBS, 0.1 mg / mL of streptomycin, 100 units of penicillin, 0.6 pg / mL of Kanamycin sulfate and 1x non-essential amino acids) were added to each well of the HTS T ranswell (Cat. No. 3391 , Corning Corporation) insert and reservoir, respectively. The HTS Transwell plates were incubated at 37 °C, 5% CO2 for 1 hour before cell seeding. CaCo-2 cells (HTB-37, American Type Culture Collection) were diluted to 6.86 x 105cells / mL with culture medium and 50 pL of cell suspension were dispensed into the filter well of the 96-well HTS Transwell plate. Cells were cultivated for 14-18 days in a cell culture incubator at 37 °C, 5% CO2, 95% relative humidity. Cell culture medium was replaced every other day, beginning no later than 24 hours after initial plating. T ransepithelial electrical resistance (TEER) values were determined using a Millicell Epithelial Volt-Ohm measuring system (Millipore). Any monolayer with a TEER value < 230 ohms cm2was discarded. Lucifer yellow leakage was also used to confirm the integrity of the monolayer, with acceptable leakage typically < 1.5%.

[0236] Stock solutions of test compounds in DMSO at 1 mM were prepared and diluted with HBSS (10 mM HEPES pH 7.4 or 10 mM MES pH 6.5) to get 5 pM working solution. To determine the rate of drug transport in the basolateral to apical direction, 125 pL of the working solution (10 mM MES pH 6.5) was added to the Transwell insert (apical compartment), and 50 pL sample was transferred immediately from the apical compartment to 200 pL of acetonitrile containing internal standard (100 nM alprazolam, 200 nM Caffeine and 100 nM tolbutamide) in a new 96-well plate as the initial donor sample (A-B). Vortexed at 1000 rpm for 10 minutes. The wells in the receiver plate (basolateral compartment) were filled with 235 pL of transport buffer (10 mM HEPES pH 7.4).

[0237] To determine the rate of drug transport in the basolateral to apical direction, 285 pL of the working solution (10 mM HEPES pH 7.4) was added to the receiver plate wells (basolateral compartment), and transfer 50 pL sample immediately from the basolateral compartment to 200 pL of acetonitrile containing IS (100 nM alprazolam, 200 nM Caffeine and 100 nM tolbutamide) in a new 96-well plate as the initial donor sample (B-A). Vortex at 1000 rpm for 10 minutes. The Transwell insert (apical compartment) was filled with 75 pL of transport buffer (10 mM MES pH 6.5). The apical to basolateral direction and the basolateral to apical direction experiments were performed at the same time, with both plates incubated at 37 °C for 2 hours.

[0238] At the end of the incubations, 50 pL samples from donor sides (apical compartment for A^B flux, and basolateral compartment for B^A) and receiver sides (basolateral compartment for A^B flux, and apical compartment for B^A) were transferred to wells of a new 96-well plate, followed by the addition of 4 volume of acetonitrile containing internal standard (100 nM alprazolam, 200 nM Caffeine and 100 nM tolbutamide). All samples were vortexed for 10 minutes, and then centrifuged at 3,220 g for 40 minutes. An aliquot of 150 pL of the supernatant was mixed with an appropriate volume of ultra-pure water before LC- MS / MS analysis to determine the concentration of compound in each sample by comparison with matrix matched standard curve. Apparent permeability (Papp) can be calculated for drug transport assays using the following equation and is reported for Examples in Table 3:Where Pappis apparent permeability (x 10'6cm / s), VA is the volume (in mL) in the acceptor well, Time is the total transport time in seconds and Area is the surface area of the membrane (0.143 cm2for Transwell-96 Well Permeable Supports).Table 3 - CaCo-2 Permeability for Selected ExamplesKey+ Papp = 0 < 1 x 10'6cm / s++ Papp > 1 s 5 x 10'6cm / s+++ Papp > 5 < 10 x 10'6cm / s++++ Papp > 10 x 10'6cm / sThus, all compounds tested showed good permeability.Oral Bioavailability

[0239] The oral bioavailability of compounds was assessed in rodent models. The experimental methods and results (Table 4) are provided hereinafter.C57BL / 6 Mouse Bioavailability, Method 1

[0240] Compounds were formulated in the stated vehicle at a concentration of 0.2 mg / mL for IV and at 0.5 mg / mL for PO. Up to three male C57BL / 6 mice per arm were dosed, which each compound being dosed at 1 mg / kg IV and 5 mg / kg PO. Blood was sampled at various timepoints and plasma isolated. 10 pL of plasma with 5 pL of blank plasma were added to 200 pL of acetonitrile containing internal standard mixture. The samples were vortexed for 30 s. After centrifugation at 4 degree Celsius, 3900 rpm for 15 min, the supernatant was diluted 3 times with water. 10 pL of diluted supernatant was injected into the LC / MS / MS system for quantitative analysis and concentration of analyte determined by comparison with matrix matched standard curve. WinNonlin (PhoenixTM, version 8.3) or other similar software may be used for pharmacokinetic calculations, including bioavailability (%F).Table 4 Oral Bioavailability (%F) of Selected ExamplesKey+ F = 0 < 5%++ F > 5 < 25%+++ F > 25 < 50%++++ F > 50%Thus, all compounds tested were orally bioavailable.

Claims

Claims1 . A compound of formula (I), or a pharmaceutically acceptable salt or N-oxide thereof:whereinL1is independently Ci-Ce-alkylene;R1is independently selected from phenyl, naphthyl, and 5-, 6-, 8-, 9- or 10- membered heteroaryl; wherein R1is optionally substituted with from 1 to 5 R5groups;R2 is independently selected from H, halo, nitro, cyano, NR7R9, OR7, SR7, SOR7, S(O)2R7, SO2NR7R7, CO2R7, C(O)R7, CONR7R7, Ci-C4-alkyl, C2-C4-alkenyl, C2-C4-alkynyl, Ci-C4- haloalkyl and cyclopropyl;R3and R4are each independently selected from H, Ci-Ce-alkyl, Ci-Ce-haloalkyl, Cs-Cs cycloalkyl, 3- to 8-membered heterocycloalkyl, Ci-C4-alkylene-R3a, C2-C4-alkylene-R3b; or wherein R3and R4, together with the nitrogen atom to which they are attached, form a 5- to 8-membered heterocycloalkyl ring, optionally substituted with 1 to 4 R6groups;R3ais independently selected from C(O)NR7R7, SO2NR7R7, CO2R7, Cs-Cs cycloalkyl, Cs-Cs- cycloalkenyl, phenyl, 3- to 8-membered heterocycloalkyl, 5- to 8-membered heterocycloalkenyl and 5- or 6-membered heteroaryl; wherein where R3ais cycloalkyl, cycloalkenyl, heterocycloalkyl or heterocycloalkenyl, R3ais optionally substituted with from 1 to 4 R6groups and where R3ais phenyl or heteroaryl, R3ais optionally substituted with from 1 to 5 R8groups;R3bis independently selected from NR7R9and OR7;R5is independently at each occurrence selected from halo, nitro, cyano, NR7R9, OR7, SR7, SOR7, S(O)2R7, S(O)2NR7R7, CO2R7, C(O)R7, C(O)NR7R7, Ci-C4-alkyl, C2-C4-alkenyl, C2-C4- alkynyl, Ci-C4-haloalkyl, phenyl, 5- or 6- membered heteroaryl, Cs-Cs cycloalkyl, Cs-Cs- cycloalkenyl, 3- to 8-membered heterocycloalkyl and 5- to 8-membered heterocycloalkenyl; wherein where R5is cycloalkyl, cycloalkenyl, heterocycloalkyl or heterocycloalkenyl, R5isoptionally substituted with from 1 to 4 R6groups and where R5is phenyl or heteroaryl, R5is optionally substituted with from 1 to 5 R8groups;R6is independently at each occurrence selected from =0, =S, halo, nitro, cyano, NR7aR9, OR7, SR7, SOR7, S(O)2R7, SO2NR7aR7a, CO2R7, C(O)R7, CONR7aR7a, Ci-C4-alkyl, C2-C4- alkenyl, C2-C4-alkynyl, Ci-C4-haloalkyl and cyclopropyl;R7and R7aare each independently at each occurrence selected from H and Ci-C4alkyl; or two R7groups, together with the nitrogen atom to which they are attached, form a 5- to 8- membered heterocycloalkyl ring, optionally substituted with 1 to 4 R6groups;R8is independently at each occurrence selected from halo, nitro, cyano, NR7R9, OR7, SR7, SOR7, S(O)2R7, SO2NR7R7, CO2R7, C(O)R7, CONR7R7, Ci-C4-alkyl, C2-C4-alkenyl, C2-C4- alkynyl, Ci-C4-haloalkyl and cyclopropyl;R9is independently at each occurrence selected from H, Ci-C4alkyl and C(O)-Ci-C4-alkyl; or R7and R9, together with the nitrogen atom to which they are attached, form a 5- to 8- membered heterocycloalkyl ring, optionally substituted with 1 to 4 R6groups; wherein any of the aforementioned alkyl, alkylene, alkenyl, or cyclopropyl groups is optionally substituted, where chemically possible, by 1 to 5 substituents which are each independently at each occurrence selected from the group consisting of: Ci-C4-alkyl, oxo, halo, nitro, cyano, NRaRb, 0Ra, SRa, CO2Ra, C(O)Ra, CONRaRa, S(O)Ra, and S(O)2Ra; wherein Rais independently at each occurrence selected from H, and Ci-C4-alkyl; and Rbis independently at each occurrence selected from H, Ci-C4-alkyl, C(O)-Ci-C4-alkyl and S(O)2-Ci-C4-alkyl.

2. A compound of claim 1 , wherein R1has the structure:wherein X1is independently selected from N and CR5b; X2is independently selected from N and CR5c; R5band R5care each independently selected from H, halo, nitro, cyano, NR7R9, OR7, SR7, SOR7, S(O)2R7, S(O)2NR7R7, CO2R7, C(O)R7, C(O)NR7R7, Ci-C4-alkyl, C2-C4- alkenyl, C2-C4-alkynyl, Ci-C4-haloalkyl; and m is an integer from 0 to 3.

4. A compound of claim 3 wherein, X2is N.

5. A compound of claim 3, wherein X2is CR5c.

6. A compound of claim 5 wherein R5cis CN.

7. A compound of any one of claims 3 to 6, wherein X1is N.

8. A compound of any one of claims 3 to 6, wherein X1is CH.

9. A compound of any one of claims 1 to 8, wherein R2is H.

10. A compound of any one of claims 1 to 9, wherein L1is -C2-alkylene, e.g. -CH2CH2-.

11. A compound of any one of claims 1 to 10, wherein R4is selected from H, Ci-Ce- alkyl and Ci-Ce-haloalkyl.

12. A compound of any one of claims 1 to 11 , wherein R3is independently selected from Ci-Ce-alkyl, Ci-Ce-haloalkyl, Cs-Cs cycloalkyl, 3- to 8-membered heterocycloalkyl, Ci-C4-alkylene-R3a, C2-C4-alkylene-R3b.

13. A compound of any one of claims 1 to 11 , wherein R3and R4are each Ci-C4-alkyl.

14. A compound of any one of claims 1 to 10, wherein R3and R4, together with the nitrogen atom to which they are attached, form a 5- to 8-membered heterocycloalkyl ring, optionally substituted with 1 to 4 R6groups.

15. A compound according to claim 1 , wherein the compound of formula (I) is selected from:

16. A pharmaceutical composition comprising a compound according to any one of claims 1 to 15, and one or more pharmaceutically acceptable excipients.

17. A compound according to any one of claims 1 to 15, or a pharmaceutical composition according to claim 16, for use as a medicament.

18. A compound according to any one of claims 1 to 15, or a pharmaceutical composition according to claim 16, for use in the treatment of: a disease of the musculoskeletal system, a disease of the skin, a metabolic disease, a disease of the Nervous System, a Cardiovascular disease, an Endocrine disorder, a disease of the eye, a disease affecting the urogenital system, a haemic or lymphatic condition, a respiratory disease, an inflammatory or autoimmune condition, a disease of the Gastrointestinal system, a Neoplasm, cancer, or a disease or disorder selected from Amelogenesis Imperfecta, Anodontia, Odontodysplasia, Branchio-Oto-Renal Syndrome, Sotos Syndrome, and Waardenburg's Syndrome.

19. A compound according to any one of claims 1 to 15, or a pharmaceutical composition according to claim 16, for use in the treatment of a disease selected from: Recessive dystrophic epidermolysis bullosa, Junctional epidermolysis bullosa, Xeroderma pigmentosum, Netherton syndrome, Duchenne Muscular Dystrophy, Becker Muscular Dystrophy, Cystic Fibrosis, Alport Syndrome, Dravet Syndrome, Aniridia, Methylmalonic Acidemia, Colorectal Cancer, Endometrium Cancer, Breast Cancer, Ovarian Cancer, Lung Squamous Cell Carcinoma, Head and Neck Squamous Cell Carcinoma, Familial adenomatous polyposis, Hemophilia A, Hemophilia B, Choroideremia, Pulmonary Artery Hypertension, Ataxia telangiectasia, Shwachman-Diamond syndrome, Mucopolysaccharidosis Type I,Mucopolysaccharidosis Type VI, Mucopolysaccharidosis type III, Niemann-Pick Disease, Primary Ciliary Dyskinesia, Usher syndrome and Retinitis Pigmentosa.

20. A compound according to any one of claims 1 to 15, or a pharmaceutical composition according to claim 16, for use in a method of treating conditions or disorders which are associated with PTC mutations in a subject.

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