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WO2025036840A1 - Dérivés de pyridine ou de pyrazine utilisés en tant qu'inducteurs de collagène vii (c7) et leur utilisation en thérapie - Google Patents

Dérivés de pyridine ou de pyrazine utilisés en tant qu'inducteurs de collagène vii (c7) et leur utilisation en thérapie Download PDF

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WO2025036840A1
WO2025036840A1 PCT/EP2024/072592 EP2024072592W WO2025036840A1 WO 2025036840 A1 WO2025036840 A1 WO 2025036840A1 EP 2024072592 W EP2024072592 W EP 2024072592W WO 2025036840 A1 WO2025036840 A1 WO 2025036840A1
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ring
compound
alkyl
phenyl
amine
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Arsenio Alberto NUEDA MARÍN
Irene JOVER MOLERO
Marie-Helene LARRAUFIE
Carlos PUIG DURÁN
Fernando Larcher Laguzzi
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Almirall SA
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
    • A61K31/4427—Non condensed pyridines; Hydrogenated derivatives thereof containing further heterocyclic ring systems
    • A61K31/444—Non condensed pyridines; Hydrogenated derivatives thereof containing further heterocyclic ring systems containing a six-membered ring with nitrogen as a ring heteroatom, e.g. amrinone
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/4965—Non-condensed pyrazines
    • A61K31/497—Non-condensed pyrazines containing further heterocyclic rings
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/535—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with at least one nitrogen and one oxygen as the ring hetero atoms, e.g. 1,2-oxazines
    • A61K31/5375—1,4-Oxazines, e.g. morpholine
    • A61K31/5377—1,4-Oxazines, e.g. morpholine not condensed and containing further heterocyclic rings, e.g. timolol
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/7028—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages
    • A61K31/7034—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages attached to a carbocyclic compound, e.g. phloridzin
    • A61K31/7036—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages attached to a carbocyclic compound, e.g. phloridzin having at least one amino group directly attached to the carbocyclic ring, e.g. streptomycin, gentamycin, amikacin, validamycin, fortimicins
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
    • A61K45/06—Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca

Definitions

  • the present invention relates to novel compounds inducing collagen VII (C7) expression.
  • This invention also relates to pharmaceutical compositions containing them, processes for their preparation and their use in the treatment of several disorders.
  • diseases accounted by truncated proteins resulting from nonsense mutations include cancer, hemophilia, cystic fibrosis, recessive dystrophic epidermolysis bullosa (RDEB), junctional epidermolysis bullosa (JEB), spinal muscular atrophy, Duchenne muscular dystrophy (DMD), among many others.
  • RDEB recessive dystrophic epidermolysis bullosa
  • JEB junctional epidermolysis bullosa
  • DMD Duchenne muscular dystrophy
  • Epidermolysis bullosa is a group of rare genetic skin disease characterized by skin fragility and blistering that affects around 500,000 people worldwide.
  • the inherited disease, dystrophic epidermolysis bullosa (DEB) is caused by recessive or dominant mutations in the COL7A1 gene encoding type VII collagen (C7).
  • DEB dystrophic epidermolysis bullosa
  • C7 type VII collagen
  • Inheritance of DEB can be autosomal dominant (DDEB) or autosomal recessive (RDEB) and all cases result from CO ZA 7 mutations (Eichstadt et al 2019).
  • RDEB RDEB
  • collagen VII C7 function
  • PTCs premature termination codons
  • frameshift splicing or missense mutations appearing alone or in combinations
  • RDEB RDEB
  • the symptoms caused by RDEB include disruption of skin and mucous membrane structural and functional architecture leading to blisters, open wounds with chronic recurrent infections, inflammation, fibrosis, syndactyly, skin itch and increased risk of squamous cell carcinoma. Other systemic symptoms are mouth and gastrointestinal tract alterations and visual impairment.
  • RDEB patients suffer from constant pain, have extremely poor quality of life, and often die at a young age (Rashidghamat et al 2017).
  • the more severe GS (generalized severe) form of RDEB affects ⁇ 2-10% of patients, most of which have biallelic PTC mutations resulting in C7 loss of function (Atanasova et al 2017, Woodley et al 2017, Eichstadt 2019).
  • Other less severe forms of RDEB involving PTCs and associated with milder symptoms often result from combinations of PTCs with other types of mutations (Soro et al 2015).
  • RDEB severity not only depends on the type of COL7A1 mutations and their location but is also influenced by individual factors which can include increased levels of mediators like the pro-fibrotic TGFp or pro-inflammatory IL-6 and MCP-1 cytokines (Odorisio et al 2014) which may also influence the levels, if any, of truncated or full length C7 that may be detectable in these patients (Ortiz-Urda et al 2005, Woodley et al 2017).
  • Enhancing translational readthrough in PTC mutations is one strategy of potential treatment for RDEB patients.
  • This approach intends to take advantage of basal readthrough, a natural process that is more frequent in PTC mutations (0.01 -1 %) than in natural termination codons (NTCs) (0.001-0.1 %), due to the proximity of NTCs to the translation termination complex and surrounding sequences (Dabrowski et al 2018).
  • NTCs natural termination codons
  • NMD nonsense mediated decay
  • gentamicin is the only small molecule drug with readthrough activity that has shown activity in RDEB caused by nonsense mutations resulting in PTCs.
  • a seminal proof of concept clinical study in RDEB patients showed the efficacy of gentamicin, which restored C7 levels and improved skin wound closure after topical and intradermal administration of the drug (Woodley et al 2017).
  • the present inventors have identified novel compounds for use in the treatment of conditions or diseases susceptible to amelioration by increasing the levels of COL7, either by restoring C-terminal truncated or full length C7 protein expression.
  • novel compounds described herein are also useful for the treatment of genetic diseases caused by nonsense mutations.
  • the present invention therefore provides a compound of formula [I] or a pharmaceutically acceptable salt, or a solvate, or a A/-oxide, or a tautomer, or a stereoisomer, or an isotopically-labelled derivative thereof, for use in the treatment of a pathological condition or disease susceptible to amelioration by increase of COL7 protein levels.
  • the invention also provides the use of a compound of formula [I] or a pharmaceutically acceptable salt, or a solvate, or a A/-oxide, or a tautomer, or a stereoisomer, or an isotopically- labelled derivative thereof, for the manufacture of a medicament for the treatment of a pathological condition or disease susceptible to amelioration by increase of COL7 protein level.
  • the invention also provides a method for treating a subject with a pathological condition or disease susceptible to amelioration by increase of COL7 protein levels, which comprises administering to said subject a therapeutically effective amount of a compound of formula [I], or a pharmaceutically acceptable salt, or a solvate, or a N-oxide, or a tautomer, or a stereoisomer, or an isotopically-labelled derivative thereof.
  • the invention also provides a combination product comprising (i) at least one compound of Formula [I] or a pharmaceutically acceptable salt, or a solvate, or a N-oxide, or a tautomer, or a stereoisomer, or an isotopically-labelled derivative thereof, and (ii) one or more active ingredients having a readthrough activity.
  • the combination product may be for use in the treatment of a pathological condition or disease susceptible to amelioration by increase of COL7 protein levels.
  • the invention also provides a compound of formula [la] or a pharmaceutically acceptable salt, or a solvate, or a A/-oxide, or a tautomer, or a stereoisomer, or an isotopically-labelled derivative thereof.
  • the invention also provides a pharmaceutical composition
  • a pharmaceutical composition comprising a compound of Formula [la] or a pharmaceutically acceptable salt, or a solvate, or a A/-oxide, or a tautomer, or a stereoisomer, or an isotopically-labelled derivative thereof, in combination with a pharmaceutically acceptable diluent or carrier.
  • the invention also provides a compound or pharmaceutical composition of Formula [la] for use as a medicament.
  • the invention provides a compound of formula [II] or a pharmaceutically acceptable salt, or a solvate, or a A/-oxide, or a tautomer, or a stereoisomer, or an isotopically-labelled derivative thereof, for use in the treatment of a pathological condition or disease susceptible to amelioration by increase of COL7 protein levels.
  • the invention also provides the use of a compound of formula [II] or a pharmaceutically acceptable salt, or a solvate, or a A/-oxide, or a tautomer, or a stereoisomer, or an isotopically- labelled derivative thereof, for the manufacture of a medicament for the treatment of a pathological condition or disease susceptible to amelioration by increase of COL7 protein level.
  • the invention also provides a method for treating a subject with a pathological condition or disease susceptible to amelioration by increase of COL7 protein levels, which comprises administering to said subject a therapeutically effective amount of a compound of formula [II], or a pharmaceutically acceptable salt, or a solvate, or a N-oxide, or a tautomer, or a stereoisomer, or an isotopically-labelled derivative thereof.
  • the invention also provides a combination product comprising (i) at least one compound of Formula [II] or a pharmaceutically acceptable salt, or a solvate, or a N-oxide, or a tautomer, or a stereoisomer, or an isotopically-labelled derivative thereof, and (ii) one or more active ingredients having a readthrough activity.
  • the combination product may be for use in the treatment of a pathological condition or disease susceptible to amelioration by increase of COL7 protein levels.
  • the invention also provides a compound of formula [Ila] or a pharmaceutically acceptable salt, or a solvate, or a A/-oxide, or a tautomer, or a stereoisomer, or an isotopically-labelled derivative thereof.
  • the invention also provides a pharmaceutical composition
  • a pharmaceutical composition comprising a compound of Formula [Ila] or a pharmaceutically acceptable salt, or a solvate, or a A/-oxide, or a tautomer, or a stereoisomer, or an isotopically-labelled derivative thereof, in combination with a pharmaceutically acceptable diluent or carrier.
  • the invention also provides a compound or pharmaceutical composition of Formula [Ila] for use as a medicament. Particular aspects of the invention are set out below:
  • W is -NR W R X or -OR W , wherein R w and R x are the same or different and are selected from hydrogen and C1-C3 alkyl;
  • A is a phenyl, 5- to 7-membered heteroaryl or 5- to 7-membered heterocyclyl ring, which ring is optionally fused to phenyl, 5- to 7-membered heteroaryl, or 5- to 7- membered heterocyclyl ring; wherein ring A is unsubstituted or substituted with one, two or three substituents which are the same or different and are selected from C1-C4 alkyl, C1-C4 haloalkyl, -C1-C3 alkyl-CONR'R", -C1-C3 alkyl-COOH, -C1-C3 alkyl-C(O)-(C-i-C3 alkyl), -C1-C3 alkyl-OR', and -C1-C3 alkyl-NR'R"; and wherein R' and R" are the same or different and are selected from hydrogen and C1-C3 alkyl;
  • R a represents the group -L-Z, wherein L is a bond or C1-C3 alkylene, and Z is a 5- to 10-membered heterocyclyl or 5- to 10-membered heteroaryl ring; wherein said ring is unsubstituted or substituted with one, two or three substituents which are the same or different and are selected from C1-C4 alkyl and C1-C4 haloalkyl; n is 0, 1 or 2; and ring B is Ce-Cw carbocyclyl which is unsubstituted or substituted with one, two, or three substituents which are the same or different and are selected from -OR"', -NR'''R' V , C1-C4 alkyl and C1-C4 haloalkyl; and wherein R"' and R iv are the same or different and are selected from hydrogen and C1-C3 alkyl; and wherein in Formula [II]:
  • W’ is -NR y R z or -OR Z , wherein R y and R z are the same or different and are selected from hydrogen and C1-C3 alkyl;
  • ring A’ is Ce-Cw aryl, 5- to 10-membered heteroaryl, or 5- to 10-membered heterocyclyl, wherein ring A’ is unsubstituted or substituted with one, two or three substituents which are the same or different and are selected from halogen, C1- C4 alkyl, C1-C4 haloalkyl, -C1-C3 alkyl-OR v , and -C1-C3 alkyl-NR v R vi ; wherein R v and R vi are the same or different and are selected from hydrogen and C1-C3 alkyl; and ring B’ is 5- to 10-membered heteroaryl, or 5- to 10-membered heterocyclyl, wherein ring B’ is unsubstituted or substituted with one, two or three substituents which are the same or different and are selected from halogen, C1-C4 alkyl, C1-C4 haloalkyl,
  • the compound for use of aspect 1 wherein the compound is a compound of Formula [I], or a pharmaceutically acceptable salt, or a solvate, or a A/-oxide, or a tautomer, or a stereoisomer, or an isotopically-labelled derivative thereof.
  • ring A is phenyl or 5- to 7-membered heterocyclyl ring, which ring is optionally fused to phenyl or 5- to 7-membered heteroaryl.
  • ring A is phenyl or 5- to 6-membered heterocyclyl ring, which ring is optionally fused to 5- to 6- membered heteroaryl.
  • ring A is phenyl, indolyl, indazolyl, or [1 ,2,4]triazolo[4,3-a]pyridinyl.
  • ring A is unsubstituted or substituted with one or two substituents which are the same or different and are selected from C1-C4 alkyl, C1-C4 haloalkyl, -C1-C3 alkyl-CONR'R'', -C1-C3 alkyl-OR', and -C1-C3 alkyl-NR'R".
  • Z is monocyclic 5- to 7-membered heterocyclyl or monocyclic 5- to 7-membered heteroaryl ring, wherein said ring contains one or two heteroatoms each independently selected from O and N.
  • COL7 deficiency condition or disease is selected from Dominant dystrophica Epidermolysis bullosa (DDEB), Recessive dystrophic epidermolysis bullosa (RDEB), Epidermolysis bullosa dystrophica pretibial, Epidermolysis bullosa pruriginosa (EBP), Nonsyndromic congenital nail disorder 8 (NDNC8), Epidermolysis bullosa dystrophica with subcorneal cleavage (EBDSC), Epidermolysis bullosa dystrophica Bart type (B-DEB), Epidermolysis bullosa dystrophica pretibial type (PR-DEB) Epidermolysis bullosa with congenital localized absence of skin and deformity of nails and Transient bullous dermolysis of the newborn (TBDN).
  • DDEB Dominant dystrophica Epidermolysis bullosa
  • RDEB Recessive dystrophic epidermolysis bullosa
  • COL7 deficiency condition or disease is Recessive dystrophic epidermolysis bullosa (RDEB).
  • RDEB Recessive dystrophic epidermolysis bullosa
  • a method for treating a subject with a pathological condition or disease susceptible to amelioration by increase of COL7 protein levels which comprises administering to said subject a therapeutically effective amount of a compound, or a pharmaceutically acceptable salt, or a solvate, or a A/-oxide, or a tautomer, or a stereoisomer, or an isotopically-labelled derivative thereof, as defined in any one of aspects 1 to 30.
  • a combination product comprising (i) at least one compound or a pharmaceutically acceptable salt, or a solvate, or a A/-oxide, or a tautomer, or a stereoisomer, or an isotopically-labelled derivative thereof as defined in any one of aspects 1 to 30, and (ii) one or more active ingredients having a readthrough activity.
  • the combination product of aspect 36 wherein the one or more active ingredients having a readthrough activity is an aminoglycoside or a macrolide.
  • the combination product of aspect 37, wherein the aminoglycoside is selected from gentamycin, G418, paromomycin, Amikacin.
  • W, ring A, R a , and n are as defined in any one of aspects 1 to 30; and ring B is adamantyl or bicyclo[2.2.2]octanyl which is unsubstituted or substituted with one, two, or three substituents which are the same or different and are selected from -OR"', -NR'i'R' , C1-C4 alkyl and C1-C4 haloalkyl; and wherein R'" and R iv are the same or different and are selected from hydrogen and C1-C3 alkyl; and wherein in Formula [Ila]:
  • W’ and ring A’ are as defined in any one of aspects 1 to 30; and ring B’ is 5- to 7-membered heteroaryl, or 5- to 10-membered heterocyclyl, wherein ring B’ is unsubstituted or substituted with one, two or three substituents which are the same or different and are selected from halogen, C1-C4 alkyl, C1-C4 haloalkyl, -C1-C3 alkyl-OR vii , and -C1-C3 alkyl-NR vii R viii ; wherein R vii and R viii are the same or different and are selected from hydrogen and C1-C3 alkyl; provided that the compound of Formula [la] is not:
  • a compound according to aspect 40 wherein the compound is a compound of Formula [Ila] or a pharmaceutically acceptable salt, or a solvate, or a A/-oxide, or a tautomer, or a stereoisomer, or an isotopically-labelled derivative thereof.
  • ring A is phenyl, monocyclic 6- to 7-membered heteroaryl, or monocyclic 6- to 7-membered heterocyclyl, preferably ring A is phenyl or pyrid inyl.
  • A’ is:
  • a pharmaceutical composition comprising a compound or a pharmaceutically acceptable salt, or a solvate, or a A/-oxide, or a tautomer, or a stereoisomer, or an isotopically-labelled derivative thereof as defined in any one of aspects 40 to 48, in combination with a pharmaceutically acceptable diluent or carrier.
  • Figure 1 a, b. Quantification of total (truncated + full length) C7 in RDEB cells (RDEB_L1 ) and recombinant HEK293 R578X cells using AlphaLISA (% read-through: C7 increase vs basal AlphaLISA levels after 48h in the presence of the indicated compound) (25 ng/mL of TGFp was added in the case of RDEB_L1 ).
  • c Detection of truncated and full length C7 after treatment with aminoglycoside reference compounds gentamicin (400 pg/mL, 0.83 mM) and G418 (25 pM) by western blot in RDEB_L1 cells using a polyclonal anti-C7 antibody
  • d Detection of truncated and full length C7 after treatment with aminoglycoside reference compounds gentamicin (2 mg/mL, 4.18 mM) and G418 (10 pM) by western blot in HEK293 R578X cells using a polyclonal anti-C7 antibody.
  • NHK normal human primary keratinocytes
  • Figure 2 a. AlphaLISA dose response evaluation of EXAMPLE 1 and EXAMPLE 7 in RDEB_L1 and HEK293 R578X cell lines, b: Analysis by western blot of EXAMPLE 1 and EXAMPLE 7 compounds in RDEB_L1 and HEK293 R578X cells at the indicated compound concentrations.
  • Figure 3 a, b. AlphaLISA dose response confirmation of activity at 48 h vs 2 h for EXAMPLE 1 and EXAMPLE 7 in the presence of 25 ng/mL of TGFp
  • FIG. 4 Comparative qPCR analysis of the induction of COL7A1 mRNA in RDEB_L1 by EXAMPLE 1 in the presence of TGFp (24h). b Immunofluorescence detection of C7 in RDEB_L1 cells (truncated C7) and NHK wild type keratinocytes (full length C7) treated with vehicle or EXAMPLE 1 in the presence of 25 ng/mL of TGFp.
  • Figure 5 a. Synergistic effect resulting from the combination of EXAMPLE 1 at suboptimal concentrations and gentamicin, in the presence of 25 ng/mL of TGFp. b. Shift in EXAMPLE 1 dose response curves in RDEB_L1 cells in the presence of increasing concentrations of gentamicin, c Western blot analysis of treated extracts with the indicated combinations of gentamicin and EXAMPLE 1.
  • terapéuticaally effective amount refers to an amount sufficient to effect treatment when administered to a patient in need of treatment.
  • treatment refers to the treatment of a disease or medical condition in a human patient which includes:
  • pathological condition or disease susceptible to amelioration by increase of C7 protein levels includes all disease states and/or conditions that are acknowledged now, or that are found in the future, to be associated with an deficiency of C7 protein levels.
  • disease states include, but are not limited to, a dermatological disease, a respiratory disease, an allergic disease, an inflammatory or autoimmune-mediated disease, a function disorder, a neurological disorder, a cardiovascular disease, a viral infection, a metabolism/endocrine function disorder, a neurological disorder, pain, bone marrow and organ transplant rejection, myelo-dysplastic syndrome, a myeloproliferative disorder (MPDs), cancer, an hematologic malignancy, leukemia, lymphoma and solid tumor.
  • MPDs myeloproliferative disorder
  • salt refers to a salt prepared from a base or acid which is acceptable for administration to a patient, such as a mammal.
  • Such salts can be derived from pharmaceutically-acceptable inorganic or organic bases and from pharmaceutically-acceptable inorganic or organic acids.
  • a N-oxide is formed from the tertiary basic amines or imines present in the molecule, using a convenient oxidising agent.
  • tautomer means two or more forms or isomers of an organic compound that readily could be interconverted into each other via a common chemical reaction called tautomerization. This reaction commonly results in the formal migration of a hydrogen atom or proton, accompanied by a switch of a single bond and adjacent double bond.
  • the concept of tautomerizations is called tautomerism. Because of the rapid interconversion, tautomers are generally considered to be the same chemical compound. In solutions in which tautomerization is possible, a chemical equilibrium of the tautomers will be reached. The exact ratio of the tautomers depends on several factors, including temperature, solvent and pH.
  • the compounds of the invention may exist in both unsolvated and solvated forms.
  • solvate is used herein to describe a molecular complex comprising a compound of the invention and an amount of one or more pharmaceutically acceptable solvent molecules.
  • hydrate is employed when said solvent is water.
  • solvate forms include, but are not limited to, compounds of the invention in association with water, acetone, dichloromethane, 2-propanol, ethanol, methanol, dimethylsulfoxide (DMSO), ethyl acetate, acetic acid, ethanolamine, or mixtures thereof.
  • the invention also includes isotopically-labelled compounds of the invention, wherein one or more atoms is replaced by an atom having the same atomic number, but an atomic mass or mass number different from the atomic mass or mass number usually found in nature.
  • isotopes suitable for inclusion in the compounds of the invention include isotopes of hydrogen, such as 2 H and 3 H, carbon, such as 11 C, 13 C and 14 C, chlorine, such as 36 CI, fluorine, such as 18 F, iodine, such as 123 l and 125 l, nitrogen, such as 13 N and 15 N, oxygen, such as 15 O, 17 O and 18 O, phosphorus, such as 32 P, and sulfur, such as 35 S.
  • Preferred isotopically- labelled compounds include deuterated derivatives of the compounds of the invention.
  • deuterated derivative embraces compounds of the invention where in a particular position at least one hydrogen atom is replaced by deuterium.
  • Deuterium (D or 2 H) is a stable isotope of hydrogen which is present at a natural abundance of 0.015 molar %.
  • Isotopically-labelled compounds derivatives of the invention can generally be prepared by conventional techniques known to those skilled in the art or by processes analogous to those described herein, using an appropriate isotopically-labelled reagent in place of the nonlabelled reagent otherwise employed.
  • alkyl or “alkylene” is intended to include both branched and straight-chain saturated aliphatic hydrocarbon groups having the specified number of carbon atoms.
  • C1-C4 alkyl or alkylene
  • Ci Ci
  • C2 and C3 alkyl groups or alkylene
  • C1-C4 alkyl denotes alkyl having 1 to 4 carbon atoms.
  • C1-C3 alkyl denotes alkyl having 1 to 3 carbon atoms. Examples of alkyl include, but are not limited to, methyl, ethyl, n- propyl, i-propyl, n-butyl, i-butyl, sec-butyl and t-butyl.
  • haloalkyl is intended to include both branched and straight-chain saturated aliphatic hydrocarbon groups having the specified number of carbon atoms, and which contains at least one halogen atom.
  • the halogen is typically chlorine, fluorine, bromine or iodine, and is preferably chlorine, fluorine or bromine, more preferably chlorine or fluorine, and most preferably fluorine.
  • aryl (or “arylene”) is intended to mean an aromatic moiety containing, if specified, the specified number of carbon atoms.
  • Ce-Cw aryl is preferably phenyl or naphthyl.
  • Ce-Cw aryl is phenyl.
  • carbocyclyl refers to a monocyclic or polycyclic (e.g., bicyclic or tricyclic), non-aromatic, saturated or partially unsaturated ring system whose ring atoms consist of carbon atoms.
  • the carbocyclyl is a saturated ring system (i.e. in which all ring bonds between adjacent carbon atoms are single bonds).
  • a polycyclic carbocyclyl may comprise fused rings (where a first ring and a second ring share two adjacent ring carbon atoms), bridged rings (where two non-adjacent ring carbon atoms of a first ring are shared with a second ring) and/or spirocyclic rings (where a first ring shares only a single ring carbon atom with a second ring).
  • a tricyclic or higher order polycyclic cycloalkyl may comprise a mixture of fused, bridged and spirocyclic relationships between its constituent rings.
  • the carbocyclyl is a bridged carbocyclyl.
  • a typical carbocyclyl is Ce-Cw carbocyclyl, preferably a bridged Cs or Cw carbocyclyl.
  • Example carbocyclyl groups include, but are not limited to, cyclohexyl, adamantyl, bicyclo[2.2.2]octanyl and the like.
  • heterocyclyl refers to a monocyclic or polycyclic (e.g., bicyclic or tricyclic), non-aromatic, saturated or partially unsaturated ring system whose ring atoms consist of carbon atoms and at least one heteroatom selected from O, S and N.
  • the at least one heteroatom is commonly 1 -3 heteroatoms, for instance 1-2 heteroatoms or 1 heteroatom.
  • the at least one heteroatom ring atom is selected from N and O.
  • the heterocyclyl is monocyclic. In one embodiment, the heterocyclyl is saturated.
  • the heterocyclyl is unsaturated.
  • Typical examples of heterocycles include 5- to 10-membered heterocycles, wherein “X- membered” means that the total number of ring atoms possessed by the heterocycle is X.
  • Preferred heterocycles are 5- to 7-membered heterocycles, 7- to 10-membered heterocycles, and 9- to 10-membered heterocycles.
  • heterocycles include, but are not limited to, azetidinyl, pyrrolidinyl, piperadinyl, oxetanyl, tetrahydrofuranyl, tetrahydropyranyl, [1 ,2,4]triazolo[4 ,3-a]pyrid inyl, 8-azabicyclo[3.2.1]octanyl, 3-azabicyclo[3.2.1]octanyl, tetrahydrothiophenyl, tetrahydrothio-pyranyl, dihydropyridine, and morpholinyl.
  • heteroaryl refers to a monocyclic or polycyclic (e.g., bicyclic or tricyclic), aromatic ring system whose ring atoms consist of carbon atoms and at least one heteroatom selected from O, S and N.
  • the at least one heteroatom is commonly 1- 3 heteroatoms, for instance 1-2 heteroatoms or 1 heteroatom.
  • the at least one heteroatom ring atom is selected from N and O. More preferably, the at least one heteroatom ring atom is N.
  • the heteroaryl is monocyclic or bicyclic, more preferably monocyclic.
  • heteroaryls include 5- to 10-membered heteroaryls, wherein “X- membered” means that the total number of ring atoms possessed by the heteroaryl is X.
  • Preferred heteroaryls are 5- to 7-membered heteroaryls, 5- to 6-membered heteroaryls, 6- to 7-membered heteroaryls, 7- to 10-membered heteroaryls, and 9- to 10-membered heteroaryls.
  • heteroaryl groups include, without limitation, furanyl, thiophenyl, imidazolyl, thiazolyl, isothiazolyl, indolyl, pyrrolyl (pyrryl), oxazolyl, indazolyl, isoxazolyl, pyrazolyl, oxadiazolyl, 1 ,2,3-oxadiazolyl, 1 ,2,4-oxadiazolyl, 1 ,2,5-oxadiazolyl, 1 ,3,4-oxadiazolyl, thiadiazolyl, 1 ,2,3-thiadiazolyl, 1 ,2,4-thiadiazolyl, 1 ,2,5-thiadiazolyl, 1 ,3,4-thiadiazolyl, triazolyl, 1 ,2,3-triazolyl, 1 ,2,4-triazolyl, 1 ,2,5-triazolyl, 1 ,3,4-triazolyl, pyridinyl, pyrrol
  • the compound for use of the invention is of formula [I] or a pharmaceutically acceptable salt, or a solvate, or a A/-oxide, or a tautomer, or a stereoisomer, or an isotopically-labelled derivative thereof:
  • W is -NR W R X or -OH. More preferably, W is -NH2 or -OH. Most preferably, W is -NH2.
  • ring A is phenyl or 5- to 7-membered heterocyclyl ring, which ring is optionally fused to phenyl or 5- to 7-membered heteroaryl. More preferably, ring A is phenyl or 5- to 6-membered heterocyclyl ring, which ring is optionally fused to 5- to 6-membered heteroaryl.
  • ring A comprises a heteroaryl or heterocyclyl ring which is optionally fused to another ring
  • said heteroaryl or heterocyclyl ring typically contains 1 , 2 or 3 heteroatoms independently selected from O or N.
  • said heteroaryl or heterocyclyl ring contains 1 , 2 or 3 nitrogen heteroatoms.
  • ring A is phenyl, indolyl, indazolyl, or [1 ,2,4]triazolo[4,3-a]pyridinyl.
  • ring A is unsubstituted or substituted with one, two or three substituents which are the same or different and are selected from C1-C4 alkyl, C1-C4 haloalkyl, -C1-C3 alkyl-CONR'R'', -C1-C3 alkyl-OR', and -C1-C3 alkyl-NR'R". More preferably, ring A is unsubstituted or substituted with one, two or three substituents which are the same or different and are selected from C1-C4 alkyl, -C1-C3 alkyl-CONR'R", and -C1-C3 alkyl-OR'.
  • ring A is unsubstituted or substituted with one or two substituents which are the same or different and are selected from the substituents defined above. More preferably, ring A is unsubstituted or substituted with one substituent which is selected from the substituents defined above.
  • ring A is unsubstituted or substituted with one or two substituents which are the same or different and are selected from C1-C4 alkyl, C1-C4 haloalkyl, -C1-C3 alkyl-CONR'R", -C1-C3 alkyl-COOH, -C1-C3 alkyl-C(O)-(Ci-C3 alkyl), -C1-C3 alkyl-OR', and -C1-C3 alkyl-NR'R".
  • ring A is unsubstituted or substituted with one or two substituents which are the same or different and are selected from C1-C4 alkyl, C1-C4 haloalkyl, -C1-C3 alkyl-CONR'R", -C1-C3 alkyl-OR', and -C1-C3 alkyl-NR'R". More preferably, ring A is unsubstituted or substituted with one substituent which is selected from -C1-C4 alkyl, -C1-C3 alkyl-CONR'R'', and -C1-C3 alkyl-OR'. Most preferably, ring A is unsubstituted or substituted with one substituent selected from propyl, -CH2-CONH2, and -CH2CH2-OH.
  • the substituent on ring A is C1-C4 alkyl
  • the substituent is C1-C3 alkyl.
  • the substituent is propyl, more preferably iso-propyl.
  • the substituent on ring A is C1-C4 haloalkyl
  • the substituent is C1-C3 haloalkyl.
  • the substituent is C1-C3 fluoroalkyl, more preferably C2-C3 fluoroalkyl.
  • the substituent on ring A is -C1-C3 alkyl-CONR'R
  • the substituent is -C1-C2 alkyl-CONR'R
  • the substituent is -C1-C2 alkyl-CONFh and more preferably is -CH2-CONH2.
  • the substituent on ring A is -C1-C3 alkyl-COOH
  • the substituent is -C1-C2 alkyl-COOH.
  • the substituent is -CH2-COOH.
  • the substituent on ring A is -C1-C3 alkyl-C(O)-(Ci-C3 alkyl)
  • the substituent is -C1-C2 alkyl-C(O)-(Ci-C2 alkyl).
  • the substituent is -CH2-C(O)-(CI-C2 alkyl), and more preferably is -CH2-C(O)-CH3.
  • the substituent on ring A is -C1-C3 alkyl-OR'
  • the substituent is -C1-C2 alkyl-OR'.
  • the substituent is -C1-C2 alkyl-OCHs or -C1-C2 alkyl-OH. More preferably, the substituent is -C1-C2 alkyl-OH, and most preferably is -CH2CH2-OH.
  • the substituent on ring A is -C1-C3 alkyl-NR'R
  • the substituent is -C1-C2 alkyl-NR'R
  • the substituent is -C1-C2 alkyl-NH2, and more preferably is -CH2CH2-NH2.
  • ring A is phenyl, indolyl, indazolyl, or [1 ,2,4]triazolo[4,3-a]pyridinyl, wherein ring A is unsubstituted or substituted with one substituent selected from propyl, -CH2-CONH2, and -CH2CH2-OH.
  • L is a bond or C1-C2 alkylene. More preferably, L is a bond or methylene. Where the formula [I] contains more than one L moiety, each L may be the same or different.
  • Z is monocyclic 5- to 7-membered heterocyclyl or monocyclic 5- to 7-membered heteroaryl ring. More preferably, Z is monocyclic 5- to 7-membered heterocyclyl or monocyclic 5- to 7-membered heteroaryl ring, wherein said ring contains one or two heteroatoms each independently selected from O and N. Yet further preferably, Z is selected from pyrazolyl, imidazolyl, pyrrolyl, furanyl, oxazolyl, piperidinyl, and morpholinyl. Most preferably, Z is selected from pyrazolyl and morpholinyl. Where the compound of the invention contains more than one Z moiety, each Z may be the same or different.
  • Z is unsubstituted or substituted with one, two or three substituents which are the same or different and are selected from C1-C4 alkyl.
  • Z is unsubstituted or substituted with one or two substituents which are the same or different and are selected from
  • Z is unsubstituted or substituted with one substituent which is C1-C4 alkyl. More preferably, Z is unsubstituted or substituted with one substituent which is methyl.
  • Z is selected from pyrazolyl and morpholinyl, wherein Z is unsubstituted or substituted with one substituent which is methyl.
  • n 0 or n is 2.
  • ring A is:
  • ring B is Cs-Cw carbocyclyl, more preferably a fused Cs-Cw carbocyclyl. Most preferably, ring B is adamantyl or bicyclo[2.2.2]octanyl.
  • Ring B is unsubstituted or substituted with one, two, or three substituents which are the same or different and are selected from -OR"' and -NR iii R iv . More preferably, ring B is unsubstituted or substituted with one, two, or three substituents which are the same or different and are -OH or -NH2. Most preferably, ring B is unsubstituted or substituted with one, two, or three substituents which are -OH.
  • ring B is unsubstituted or substituted with one or two substituents which are the same or different and are selected from the substituents defined above. More preferably, ring B is unsubstituted or substituted with one substituent which is selected from the substituents defined above. Most preferably, ring B is substituted with one substituent which is selected from the substituents defined above.
  • ring B is unsubstituted or substituted with one or two substituents which are the same or different and are selected from -OR"' and -NR'''R' V .
  • ring B is unsubstituted or substituted with one substituent selected from -OR'" and -NR'''R' V .
  • ring B is substituted with one substituent which is -OR"'.
  • ring B is substituted with one substituent which is -OH.
  • the substituent on ring B is -OR"'
  • the substituent is -OCH3 or -OH.
  • the substituent is -OH.
  • the substituent on ring B is -NR iii R iv
  • the substituent is -NHCH3 or -NH2.
  • the substituent is -NH2.
  • the substituent on ring B is C1-C4 alkyl
  • the substituent is C1-C3 alkyl or C1-C2 alkyl.
  • the substituent is methyl.
  • the substituent on ring B is C1-C4 haloalkyl
  • the substituent is C1-C3 haloalkyl or C1-C2 haloalkyl.
  • the substituent is -CF3.
  • ring B is adamantyl or bicyclo[2.2.2]octanyl which is substituted with one substituent which is -OH.
  • ring B is:
  • - W is -NH 2 ;
  • ring A is phenyl or 5- to 7-membered heterocyclyl ring, which ring is optionally fused to phenyl or 5- to 7-membered heteroaryl, wherein ring A is unsubstituted or substituted with one or two substituents which are the same or different and are selected from Ci- C4 alkyl, C1-C4 haloalkyl, -C1-C3 alkyl-CONR'R'', -C1-C3 alkyl-OR', and -C1-C3 alkyl-NR'R";
  • R a represents the group -L-Z, wherein L is a bond or C1-C2 alkylene, and Z is 5- to 7- membered heterocyclyl or 5- to 7-membered heteroaryl ring, wherein said ring contains one or two heteroatoms each independently selected from O and N, and wherein Z is unsubstituted or substituted with one, two or three substituents which are the same or different and are selected from C1-C4 alkyl; ring B is Cs-Cw carbocyclyl, which is unsubstituted or substituted with one substituent selected from -OR"' and -NR iii R iv ; and
  • R', R", R"', R iv and n are as defined for Formula [I],
  • - W is -NH 2 ;
  • ring A is phenyl or 5- to 6-membered heterocyclyl ring, which ring is optionally fused to 5- to 6-membered heteroaryl, wherein ring A is unsubstituted or substituted with one substituent which is selected from -C1-C4 alkyl, -C1-C3 alkyl-CONR'R", and -C1-C3 alkyl-OR';
  • R a represents the group -L-Z, wherein L is a bond or methylene, and Z is 5- to 7- membered heterocyclyl or 5- to 7-membered heteroaryl ring, wherein said ring contains one or two heteroatoms each independently selected from O and N, and wherein Z is unsubstituted or substituted with one substituent which is C1-C4 alkyl; ring B is Cs-Cw carbocyclyl, which is substituted with one substituent which is -OR'"; and
  • R', R", R"', R' v and n are as defined for Formula [I], In a yet further preferred embodiment of Formula [I]:
  • W is -NH 2 ;
  • ring A is phenyl, indolyl, indazolyl, or [1 ,2,4]triazolo[4,3-a]pyridinyl, wherein ring A is unsubstituted or substituted with one substituent selected from propyl, -CH2-CONH2, and -CH2CH2-OH;
  • R a represents the group -L-Z, wherein L is a bond or methylene, and Z is 5- to 7- membered heterocyclyl or 5- to 7-membered heteroaryl ring, wherein said ring contains one or two heteroatoms each independently selected from O and N, and wherein Z is unsubstituted or substituted with one substituent which is methyl; ring B is adamantyl or bicyclo[2.2.2]octanyl which is substituted with one substituent which is -OH; and n is as defined for Formula [I],
  • Preferred compounds of Formula [I] are:
  • the invention also provides a compound of formula [la] or a pharmaceutically acceptable salt, or a solvate, or a A/-oxide, or a tautomer, or a stereoisomer, or an isotopically-labelled derivative thereof:
  • W is -NR W R X or -OH. More preferably, W is -NH2 or -OH. Most preferably, W is -NH2.
  • ring A is phenyl or 5- to 7-membered heterocyclyl ring, which ring is optionally fused to phenyl or 5- to 7-membered heteroaryl. More preferably, ring A is phenyl or 5- to 6-membered heterocyclyl ring, which ring is optionally fused to 5- to 6- membered heteroaryl.
  • ring A comprises a heteroaryl or heterocyclyl ring which is optionally fused to another ring
  • said heteroaryl or heterocyclyl ring typically contains 1 , 2 or 3 heteroatoms independently selected from O or N.
  • said heteroaryl or heterocyclyl ring contains 1 , 2 or 3 nitrogen heteroatoms.
  • ring A is phenyl, indolyl, indazolyl, or [1 ,2,4]triazolo[4,3-a]pyridinyl.
  • ring A is unsubstituted or substituted with one, two or three substituents which are the same or different and are selected from C1-C4 alkyl, C1-C4 haloalkyl, -C1-C3 alkyl-CONR'R'', -C1-C3 alkyl-OR', and -C1-C3 alkyl-NR'R". More preferably, ring A is unsubstituted or substituted with one, two or three substituents which are the same or different and are selected from C1-C4 alkyl, -C1-C3 alkyl-CONR'R", and -C1-C3 alkyl-OR'.
  • ring A is unsubstituted or substituted with one or two substituents which are the same or different and are selected from the substituents defined above. More preferably, ring A is unsubstituted or substituted with one substituent which is selected from the substituents defined above.
  • ring A is unsubstituted or substituted with one or two substituents which are the same or different and are selected from C1-C4 alkyl, C1-C4 haloalkyl, -C1-C3 alkyl-CONR'R", -C1-C3 alkyl-COOH, -C1-C3 alkyl-C(O)-(Ci-C3 alkyl), -C1-C3 alkyl-OR', and -C1-C3 alkyl-NR'R".
  • ring A is unsubstituted or substituted with one or two substituents which are the same or different and are selected from C1-C4 alkyl, C1-C4 haloalkyl, -C1-C3 alkyl-CONR'R", -C1-C3 alkyl-OR', and -C1-C3 alkyl-NR'R". More preferably, ring A is unsubstituted or substituted with one substituent which is selected from -C1-C4 alkyl, -C1-C3 alkyl-CONR'R'', and -C1-C3 alkyl-OR'. Most preferably, ring A is unsubstituted or substituted with one substituent selected from propyl, -CH2-CONH2, and -CH2CH2-OH.
  • the substituent on ring A is C1-C4 alkyl
  • the substituent is C1-C3 alkyl.
  • the substituent is propyl, more preferably iso-propyl.
  • the substituent on ring A is C1-C4 haloalkyl
  • the substituent is C1-C3 haloalkyl.
  • the substituent is C1-C3 fluoroalkyl, more preferably C2-C3 fluoroalkyl.
  • the substituent on ring A is -C1-C3 alkyl-CONR'R
  • the substituent is -C1-C2 alkyl-CONR'R
  • the substituent is -C1-C2 alkyl-CONFh and more preferably is -CH2-CONH2.
  • the substituent on ring A is -C1-C3 alkyl-COOH
  • the substituent is -C1-C2 alkyl-COOH.
  • the substituent is -CH2-COOH.
  • the substituent on ring A is -C1-C3 alkyl-C(O)-(Ci-C3 alkyl)
  • the substituent is -C1-C2 alkyl-C(O)-(Ci-C2 alkyl).
  • the substituent is -CH2-C(O)-(CI-C2 alkyl), and more preferably is -CH2-C(O)-CH3.
  • the substituent on ring A is -C1-C3 alkyl-OR'
  • the substituent is -C1-C2 alkyl-OR'.
  • the substituent is -C1-C2 alkyl-OCHs or -C1-C2 alkyl-OH. More preferably, the substituent is -C1-C2 alkyl-OH, and most preferably is -CH2CH2-OH.
  • the substituent on ring A is -C1-C3 alkyl-NR'R
  • the substituent is -C1-C2 alkyl-NR'R
  • the substituent is -C1-C2 alkyl-NH2, and more preferably is -CH2CH2-NH2.
  • ring A is phenyl, indolyl, indazolyl, or [1 ,2,4]triazolo[4,3-a]pyridinyl, wherein ring A is unsubstituted or substituted with one substituent selected from propyl, -CH2-CONH2, and -CH2CH2-OH.
  • L is a bond or C1-C2 alkylene. More preferably, L is a bond or methylene. Where the formula [I] contains more than one L moiety, each L may be the same or different.
  • Z is monocyclic 5- to 7-membered heterocyclyl or monocyclic 5- to 7-membered heteroaryl ring. More preferably, Z is monocyclic 5- to 7-membered heterocyclyl or monocyclic 5- to 7-membered heteroaryl ring, wherein said ring contains one or two heteroatoms each independently selected from O and N. Yet further preferably, Z is selected from pyrazolyl, imidazolyl, pyrrolyl, furanyl, oxazolyl, piperidinyl, and morpholinyl. Most preferably, Z is selected from pyrazolyl and morpholinyl. Where the compound of the invention contains more than one Z moiety, each Z may be the same or different.
  • Z is unsubstituted or substituted with one, two or three substituents which are the same or different and are selected from C1-C4 alkyl.
  • Z is unsubstituted or substituted with one or two substituents which are the same or different and are selected from C1-C4 alkyl. More preferably, Z is unsubstituted or substituted with one substituent which is C1-C4 alkyl. Most preferably, Z is unsubstituted or substituted with one substituent which is methyl.
  • Z is selected from pyrazolyl and morpholinyl, wherein Z is unsubstituted or substituted with one substituent which is methyl.
  • n 0 or n is 2.
  • ring A is:
  • ring B is adamantyl or bicyclo[2.2.2]octanyl, which is unsubstituted or substituted with one, two, or three substituents which are the same or different and are selected from -OR'", -NR iii R iv , C1-C4 alkyl and C1-C4 haloalkyl; and wherein R"' and R iv are the same or different and are selected from hydrogen and C1-C3 alkyl.
  • Ring B is unsubstituted or substituted with one, two, or three substituents which are the same or different and are selected from -OR"' and -NR iii R iv .
  • ring B is unsubstituted or substituted with one, two, or three substituents which are the same or different and are -OH or -NH2. Most preferably, ring B is unsubstituted or substituted with one, two, or three substituents which are -OH.
  • ring B is unsubstituted or substituted with one or two substituents which are the same or different and are selected from the substituents defined above. More preferably, ring B is unsubstituted or substituted with one substituent which is selected from the substituents defined above. Most preferably, ring B is substituted with one substituent which is selected from the substituents defined above.
  • ring B is unsubstituted or substituted with one or two substituents which are the same or different and are selected from -OR"' and -NR'''R' V .
  • ring B is unsubstituted or substituted with one substituent selected from -OR'" and -NR'''R' V .
  • ring B is substituted with one substituent which is -OR"'.
  • ring B is substituted with one substituent which is -OH.
  • the substituent on ring B is -OR"'
  • the substituent is -OCH3 or -OH.
  • the substituent is -OH.
  • the substituent on ring B is -NR'''R' V
  • the substituent is -NHCH3 or -NH2.
  • the substituent is -NH2.
  • the substituent on ring B is C1-C4 alkyl
  • the substituent is C1-C3 alkyl or C1-C2 alkyl.
  • the substituent is methyl.
  • the substituent on ring B is C1-C4 haloalkyl
  • the substituent is C1-C3 haloalkyl or C1-C2 haloalkyl.
  • the substituent is -CF3.
  • ring B is adamantyl or bicyclo[2.2.2]octanyl which is substituted with one substituent which is -OH
  • ring B is or In a preferred embodiment of Formula [la]:
  • - W is -NH 2 ;
  • ring A is phenyl or 5- to 7-membered heterocyclyl ring, which ring is optionally fused to phenyl or 5- to 7-membered heteroaryl, wherein ring A is unsubstituted or substituted with one or two substituents which are the same or different and are selected from Ci- C4 alkyl, C1-C4 haloalkyl, -C1-C3 alkyl-CONR'R'', -C1-C3 alkyl-OR', and -C1-C3 alkyl-NR'R";
  • R a represents the group -L-Z, wherein L is a bond or C1-C2 alkylene, and Z is 5- to 7- membered heterocyclyl or 5- to 7-membered heteroaryl ring, wherein said ring contains one or two heteroatoms each independently selected from O and N, and wherein Z is unsubstituted or substituted with one, two or three substituents which are the same or different and are selected from C1-C4 alkyl; ring B is adamantyl or bicyclo[2.2.2]octanyl, which is unsubstituted or substituted with one substituent selected from -OR"' and NR iii R iv .
  • R', R", R"', R iv and n are as defined for Formula [la].
  • - W is -NH 2 ;
  • ring A is phenyl or 5- to 6-membered heterocyclyl ring, which ring is optionally fused to 5- to 6-membered heteroaryl, wherein ring A is unsubstituted or substituted with one substituent which is selected from -C1-C4 alkyl, -C1-C3 alkyl-CONR'R", and -C1-C3 alkyl-OR';
  • R a represents the group -L-Z, wherein L is a bond or methylene, and Z is 5- to 7- membered heterocyclyl or 5- to 7-membered heteroaryl ring, wherein said ring contains one or two heteroatoms each independently selected from O and N, and wherein Z is unsubstituted or substituted with one substituent which is C1-C4 alkyl; ring B is adamantyl or bicyclo[2.2.2]octanyl, which is substituted with one substituent which is -OR"'; and
  • R', R", R"', R' v and n are as defined for Formula [la].
  • R', R", R"', R' v and n are as defined for Formula [la].
  • W is -NH 2 ;
  • ring A is phenyl, indolyl, indazolyl, or [1 ,2,4]triazolo[4,3-a]pyridinyl, wherein ring A is unsubstituted or substituted with one substituent selected from propyl, -CH2-CONH2, and -CH2CH2-OH;
  • R a represents the group -L-Z, wherein L is a bond or methylene, and Z is 5- to 7- membered heterocyclyl or 5- to 7-membered heteroaryl ring, wherein said ring contains one or two heteroatoms each independently selected from O and N, and wherein Z is unsubstituted or substituted with one substituent which is methyl; ring B is adamantyl or bicyclo[2.2.2]octanyl which is substituted with one substituent which is -OH; and n is as defined for Formula [la].
  • Preferred compounds of Formula [la] are:
  • the compound for use of the invention is of formula [II] or a pharmaceutically acceptable salt, or a solvate, or a A/-oxide, or a tautomer, or a stereoisomer, or an isotopically-labelled derivative thereof:
  • W’ is -NR y R z or -OH. More preferably, W’ is -NH2 or -OH. Most preferably, W’ is -NH2.
  • Y is CH.
  • ring A’ is phenyl, monocyclic 5- to 7-membered heteroaryl, or monocyclic 5- to 7-membered heterocyclyl ring. More preferably, ring A’ is phenyl or monocyclic 5- to 7-membered heteroaryl ring.
  • ring A is heteroaryl or heterocyclyl ring
  • said ring typically contains 1 , 2 or 3 heteroatoms independently selected from O or N.
  • said ring contains 1 or 2 heteroatoms independently selected from O or N.
  • said heteroaryl or heterocyclyl group contains one heteroatom which is O or N.
  • ring A is phenyl, furanyl, or pyrid inyl.
  • ring A is unsubstituted or substituted with one, two or three substituents which are the same or different and are selected from halogen, C1-C4 alkyl and C1-C4 haloalkyl. More preferably, ring A is unsubstituted or substituted with one, two or three substituents which are the same or different and are selected from halogen and C1-C4 haloalkyl. Most preferably, ring A’ is unsubstituted or substituted with one, two or three substituents which are the same or different and are halogen.
  • ring A’ is unsubstituted or substituted with one or two substituents which are the same or different and are selected from the substituents defined above. More preferably, ring A is unsubstituted or substituted with one substituent which is selected from the substituents defined above.
  • ring A is unsubstituted or substituted with one or two substituents which are the same or different and are selected from halogen, C1-C4 alkyl and C1-C4 haloalkyl.
  • ring A’ is unsubstituted or substituted with one or two substituents which are the same or different and are selected from halogen and C1-C4 haloalkyl. More preferably, ring A’ is unsubstituted or substituted with one or two substituents which are the same or different and are halogen. Most preferably, ring A’ is unsubstituted or substituted with one or two substituents which are fluorine.
  • the substituent on ring A is halogen
  • the substituent is fluorine or chlorine.
  • the substituent is fluorine.
  • the substituent on ring A’ is C1-C4 alkyl
  • the substituent is C1-C3 alkyl.
  • the substituent is C1-C2 alkyl, and more preferably is methyl.
  • the substituent on ring A’ is C1-C4 haloalkyl
  • the substituent is C1-C3 haloalkyl.
  • the substituent is C1-C3 fluoroalkyl, and more preferably is C1-C2 fluoroalkyl.
  • the substituent on ring A is -C1-C3 alkyl-OR v
  • the substituent is -C1-C2 alkyl-OR v
  • the substituent is -C1-C2 alkyl-OH, and most preferably is -CH2CH2-OH.
  • the substituent on ring A is -C1-C3 alkyl-NR v R vi
  • the substituent is -C1-C2 alkyl-NR v R vi
  • the substituent is C1-C2 alkyl-NH2, and more preferably is -CH2CH2-NH2.
  • ring A’ is phenyl, furanyl, or pyridinyl, wherein ring A’ is unsubstituted or substituted with one or two substituents which are fluorine.
  • ring A’ is:
  • ring B’ is 5- to 10-membered heteroaryl. More preferably, ring B’ is monocyclic 5- to 7-membered heteroaryl.
  • ring B’ is a heteroaryl or heterocyclyl ring which contains 1 , 2 or 3 heteroatoms independently selected from O or N.
  • said ring contains 1 or 2 heteroatoms independently selected from O or N. More preferably, said ring contains one heteroatom which is N.
  • ring B’ is pyridinyl.
  • ring B’ is unsubstituted or substituted with one, two or three substituents which are the same or different and are selected from C1-C4 alkyl and C1-C4 haloalkyl. More preferably, ring B’ is unsubstituted or substituted with one, two or three substituents which are the same or different and are selected from C1-C4 alkyl. Most preferably, ring B’ is unsubstituted or substituted with one, two or three substituents which are methyl.
  • ring B’ is unsubstituted or substituted with one or two substituents which are the same or different and are selected from the substituents defined above. More preferably, ring B’ is unsubstituted or substituted with two substituents which are selected from the substituents defined above.
  • ring B’ is unsubstituted or substituted with one or two substituents which are the same or different and are selected from C1-C4 alkyl and C1-C4 haloalkyl.
  • ring B’ is unsubstituted or substituted with one or two substituents which are the same or different and are selected from C1-C4 alkyl. More preferably, ring B’ is unsubstituted or substituted with one or two substituents which are methyl.
  • the substituent on ring B’ is halogen
  • the substituent is fluorine or chlorine.
  • the substituent is fluorine.
  • the substituent on ring B’ is C1-C4 alkyl
  • the substituent is C1-C3 alkyl
  • the substituent is C1-C2 alkyl, and more preferably is methyl.
  • the substituent on ring B’ is C1-C4 haloalkyl
  • the substituent is C1-C3 haloalkyl.
  • the substituent is C1-C3 fluoroalkyl, and more preferably is C1-C2 fluoroalkyl.
  • the substituent on ring B’ is -C1-C3 alkyl-OR vii
  • the substituent is -C1-C2 alkyl-OR vii
  • the substituent is -C1-C2 alkyl-OH, and most preferably is -CH2CH2-OH.
  • the substituent on ring B’ is -C1-C3 alkyl-NR vii R viii
  • the substituent is -C1-C2 alkyl-NR vii R viii
  • the substituent is C1-C2 alkyl-NH2, and more preferably is -CH2CH2-NH2.
  • ring B’ is pyridinyl which is unsubstituted or substituted with one or two substituents which are methyl. More preferably, ring B’ is '
  • - W’ is -NH 2 ;
  • ring A’ is phenyl, monocyclic 5- to 7-membered heteroaryl, or monocyclic 5- to 7- membered heterocyclyl, wherein ring A is unsubstituted or substituted with one or two substituents which are the same or different and are selected from halogen, C1-C4 alkyl and C1-C4 haloalkyl;
  • ring B’ is 5- to 10-membered heteroaryl, wherein ring B’ is unsubstituted or substituted with one or two substituents which are the same or different and are selected from C1- C4 alkyl and C1-C4 haloalkyl; and
  • Y is as defined for Formula [II],
  • - W’ is -NH 2 ;
  • ring A is phenyl or monocyclic 5- to 7-membered heteroaryl, wherein ring A is unsubstituted or substituted with one or two substituents which are fluorine;
  • ring B’ is monocyclic 5- to 7-membered heteroaryl, wherein ring B’ is unsubstituted or substituted with one or two substituents which are the same or different and are selected from C1-C4 alkyl; and
  • Y is as defined for Formula [II],
  • Preferred compounds of Formula [II] are:
  • the invention also provides a compound of formula [Ila] or a pharmaceutically acceptable salt, or a solvate, or a A/-oxide, or a tautomer, or a stereoisomer, or an isotopically-labelled derivative thereof:
  • W’ is -NR y R z or -OH. More preferably, W’ is -NH2 or -OH. Most preferably, W’ is -NH2.
  • Y is CH.
  • ring A’ is phenyl, monocyclic 5- to 7-membered heteroaryl, or monocyclic 5- to 7-membered heterocyclyl ring. More preferably, ring A’ is phenyl or monocyclic 5- to 7-membered heteroaryl ring.
  • ring A is heteroaryl or heterocyclyl ring
  • said ring typically contains 1 , 2 or 3 heteroatoms independently selected from O or N.
  • said ring contains 1 or 2 heteroatoms independently selected from O or N.
  • said heteroaryl or heterocyclyl group contains one heteroatom which is O or N.
  • ring A’ is phenyl, furanyl, or pyrid inyl.
  • ring A is unsubstituted or substituted with one, two or three substituents which are the same or different and are selected from halogen, C1-C4 alkyl and C1-C4 haloalkyl. More preferably, ring A is unsubstituted or substituted with one, two or three substituents which are the same or different and are selected from halogen and C1-C4 haloalkyl. Most preferably, ring A’ is unsubstituted or substituted with one, two or three substituents which are the same or different and are halogen.
  • ring A’ is unsubstituted or substituted with one or two substituents which are the same or different and are selected from the substituents defined above. More preferably, ring A is unsubstituted or substituted with one substituent which is selected from the substituents defined above.
  • ring A is unsubstituted or substituted with one or two substituents which are the same or different and are selected from halogen, C1-C4 alkyl and C1-C4 haloalkyl.
  • ring A’ is unsubstituted or substituted with one or two substituents which are the same or different and are selected from halogen and C1-C4 haloalkyl. More preferably, ring A’ is unsubstituted or substituted with one or two substituents which are the same or different and are halogen. Most preferably, ring A is unsubstituted or substituted with one or two substituents which are fluorine.
  • the substituent on ring A is halogen
  • the substituent is fluorine or chlorine.
  • the substituent is fluorine.
  • the substituent on ring A’ is C1-C4 alkyl
  • the substituent is C1-C3 alkyl.
  • the substituent is C1-C2 alkyl, and more preferably is methyl.
  • the substituent on ring A’ is C1-C4 haloalkyl
  • the substituent is C1-C3 haloalkyl.
  • the substituent is C1-C3 fluoroalkyl, and more preferably is C1-C2 fluoroalkyl.
  • the substituent on ring A’ is -C1-C3 alkyl-OR v
  • the substituent is -C1-C2 alkyl-OR v
  • the substituent is -C1-C2 alkyl-OH, and most preferably is -CH2CH2-OH.
  • the substituent on ring A is -C1-C3 alkyl-NR v R vi
  • the substituent is -C1-C2 alkyl-NR v R vi
  • the substituent is C1-C2 alkyl-NH2, and more preferably is -CH2CH2-NH2.
  • ring A’ is phenyl, furanyl, or pyridinyl, wherein ring A’ is unsubstituted or substituted with one or two substituents which are fluorine.
  • ring A’ is:
  • ring A’ is phenyl, monocyclic 6- to 7-membered heteroaryl, or monocyclic 6- to 7-membered heterocyclyl ring. More preferably, ring A’ is phenyl or monocyclic 6- to 7-membered heteroaryl ring. Most preferably, ring A’ is phenyl or pyridinyl. Most preferably, ring A’ is:
  • ring B’ is 5- to 6-membered heteroaryl, more preferably B’ is 6-membered heteroaryl.
  • ring B’ is a heteroaryl or heterocyclyl ring which contains 1 , 2 or 3 heteroatoms independently selected from O or N.
  • said ring contains 1 or 2 heteroatoms independently selected from O or N. More preferably, said ring contains one heteroatom which is N.
  • ring B’ is pyridinyl.
  • ring B’ is unsubstituted or substituted with one, two or three substituents which are the same or different and are selected from C1-C4 alkyl and C1-C4 haloalkyl. More preferably, ring B’ is unsubstituted or substituted with one, two or three substituents which are the same or different and are selected from C1-C4 alkyl. Most preferably, ring B’ is unsubstituted or substituted with one, two or three substituents which are methyl.
  • ring B’ is unsubstituted or substituted with one or two substituents which are the same or different and are selected from the substituents defined above. More preferably, ring B’ is unsubstituted or substituted with two substituents which are selected from the substituents defined above.
  • ring B’ is unsubstituted or substituted with one or two substituents which are the same or different and are selected from C1-C4 alkyl and C1-C4 haloalkyl.
  • ring B’ is unsubstituted or substituted with one or two substituents which are the same or different and are selected from C1-C4 alkyl. More preferably, ring B’ is unsubstituted or substituted with one or two substituents which are methyl.
  • the substituent on ring B’ is halogen
  • the substituent is fluorine or chlorine.
  • the substituent is fluorine.
  • the substituent on ring B’ is C1-C4 alkyl
  • the substituent is C1-C3 alkyl
  • the substituent is C1-C2 alkyl, and more preferably is methyl.
  • the substituent on ring B’ is C1-C4 haloalkyl
  • the substituent is C1-C3 haloalkyl.
  • the substituent is C1-C3 fluoroalkyl, and more preferably is C1-C2 fluoroalkyl.
  • the substituent on ring B’ is -C1-C3 alkyl-OR vii
  • the substituent is -C1-C2 alkyl-OR vii
  • the substituent is -C1-C2 alkyl-OH, and most preferably is -CH2CH2-OH.
  • the substituent on ring B’ is -C1-C3 alkyl-NR vii R viii
  • the substituent is -C1-C2 alkyl-NR vii R viii
  • the substituent is C1-C2 alkyl-NH2, and more preferably is -CH2CH2-NH2.
  • ring B’ is pyridinyl which is unsubstituted or substituted with one or two substituents which are methyl.
  • ring B’ is Most preferably, ring B’ is:
  • - W’ is -NH 2 ;
  • ring A’ is phenyl, monocyclic 5- to 7-membered heteroaryl, or monocyclic 5- to 7- membered heterocyclyl, wherein ring A is unsubstituted or substituted with one or two substituents which are the same or different and are selected from halogen, C1-C4 alkyl and C1-C4 haloalkyl;
  • ring B’ is 5- to 7-membered heteroaryl, wherein ring B’ is unsubstituted or substituted with one or two substituents which are the same or different and are selected from C1- C4 alkyl and C1-C4 haloalkyl; and
  • Y is as defined for Formula [Ila],
  • - W’ is -NH 2 ;
  • ring A is phenyl or monocyclic 5- to 7-membered heteroaryl, wherein ring A is unsubstituted or substituted with one or two substituents which are fluorine;
  • ring B’ is monocyclic 5- to 7-membered heteroaryl, wherein ring B’ is unsubstituted or substituted with one or two substituents which are the same or different and are selected from C1-C4 alkyl; and
  • Y is as defined for Formula [Ila],
  • Preferred compounds of Formula [Ila] are:
  • the compounds of the invention may be administered as part of a combination product comprising (i) at least one compound of the invention or a pharmaceutically acceptable salt, or a solvate, or a A/-oxide, or a tautomer, or a stereoisomer, or an isotopically-labelled derivative thereof, and (ii) one or more active ingredients having a readthrough activity.
  • the active ingredient having a readthrough activity may be any compound which enhances readthrough of premature termination codons (PTCs) compared to basal levels.
  • the one or more active ingredients having a readthrough activity is an aminoglycoside or a macrolide, preferably an aminoglycoside.
  • Suitable aminoglycosides include gentamycin, G418, paromomycin, Amikacin.
  • the aminoglycoside is gentamycin.
  • the compounds and combination products of the present invention may be used in the treatment of a pathological condition or disease susceptible to amelioration by increase of COL7 protein levels.
  • the pathological condition or disease is susceptible to amelioration by increase of full-length COL7 protein and/or truncated COL7 protein.
  • the pathological condition or disease susceptible to amelioration by increase of COL7 protein levels is a COL7 deficiency condition or disease.
  • the pathological condition or disease susceptible to amelioration by increase of COL7 protein levels is a COL7 deficiency condition or disease.
  • said pathological condition or disease susceptible to amelioration by increase of COL7 protein levels includes Dominant dystrophic Epidermolysis bullosa (DDEB), Recessive dystrophic epidermolysis bullosa (RDEB), Epidermolysis bullosa dystrophica pretibial, Epidermolysis bullosa pruriginosa (EBP), Nonsyndromic congenital nail disorder 8 (NDNC8), Epidermolysis bullosa dystrophica with subcorneal cleavage (EBDSC), Epidermolysis bullosa dystrophic Bart type (B-DEB), Epidermolysis bullosa dystrophic pretibial type (PR-DEB) Epidermolysis bullosa with congenital localized absence of skin and deformity of nails and Transient bullous dermolysis of the newborn (DDEB), Reces
  • the invention provides a compound of formula [I] or a pharmaceutically acceptable salt, or a solvate, or a A/-oxide, or a tautomer, or a stereoisomer, or an isotopically- labelled derivative thereof, for use in the treatment of Recessive dystrophic epidermolysis bullosa (RDEB).
  • RDEB Recessive dystrophic epidermolysis bullosa
  • the invention provides a compound of formula [II] or a pharmaceutically acceptable salt, or a solvate, or a A/-oxide, or a tautomer, or a stereoisomer, or an isotopically- labelled derivative thereof, for use in the treatment of Recessive dystrophic epidermolysis bullosa (RDEB).
  • RDEB Recessive dystrophic epidermolysis bullosa
  • the invention also provides a method for treating a nonsense-mutation-mediated genetic disease, comprising administering to said subject a therapeutically effective amount of a compound of the invention or a pharmaceutically acceptable salt, or a solvate, or a A/-oxide, or a tautomer, or a stereoisomer, or an isotopically-labelled derivative thereof, or a pharmaceutical composition or a combination product of the present invention.
  • the invention also provides a method for treating a subject diagnosed with a pathological condition or disease caused partially or entirely by a nonsense or PTC mutation.
  • pathological conditions or diseases include epidermolysis bullosa, including RDEB and JEB, cystic fibrosis (CF), Duchenne muscular dystrophy (DMD), ataxia telangiectasia, Lysosomal Storage Disorders (including Fabry disease, Niemann-Pick A/B, Gangliosidosis, Mucopolysaccharidosis (MPS) Type l-Hurler, MPS Type lll-B, MPS Type VI, Batten disease) Hemophilia, Spinal Muscular Atrophy, Hereditary Ocular Disorders (including Choroideremia, Leber’s Congenital Amaurosis, Ocular Coloboma, Retinitis Pigmentosa, Usher Syndrome or Congenital Aniridia) and epidermolysis bullosa (including RDEB, DDEB, J
  • the nonsense or PTC mutation is within a gene selected from COL7A1 , CFTR, DMD, ATM, GLA, SMPD1 , GLB1 , IDUA, IDS, NAGLU, CLN1 , CLN2, FVIII, FIX, SMN2, REP1 , RPE65, PAX2, Almbl , RHO, RP2, PCDH15, USH1C, Pax6, ARSB, or MECP2.
  • the nonsense or PTC mutation is within the COL7A1 gene.
  • the invention also provides a method for enhancing skin wound healing in a subject suffering from DEB due to a nonsense mutation in the COL7A1 gene, comprising administering to said subject a therapeutically effective amount of a compound of the invention or a pharmaceutically acceptable salt, or a solvate, or a A/-oxide, or a tautomer, or a stereoisomer, or an isotopically-labelled derivative thereof.
  • the invention also provides a method for inducing expressing of full-length and/or truncated Col7 protein in a subject, comprising administering to said subject a therapeutically effective amount of a compound of the invention or a pharmaceutically acceptable salt, or a solvate, or a A/-oxide, or a tautomer, or a stereoisomer, or an isotopically-labelled derivative thereof.
  • the invention also provides a method for treating a subject with a pathological condition or disease susceptible to amelioration by increase of COL7 protein levels which comprises administering to said subject a therapeutically effective amount of a compound of the invention or a pharmaceutically acceptable salt, or a solvate, or a A/-oxide, or a tautomer, or a stereoisomer, or an isotopically-labelled derivative thereof.
  • the invention also provides a compound of the invention for use as a medicament.
  • compositions and combination products described herein may also be used in the uses and methods described above.
  • the compounds of the invention when used in a method of treatment, can be administered alone, but generally will be administered with a pharmaceutical carrier selected on the basis of the chosen route of administration and standard pharmaceutical practice.
  • the compounds of the present invention can be included in a pharmaceutical composition.
  • the pharmaceutical composition is suitable for oral administration.
  • a solid dosage form suitable for oral administration e.g., a tablet, capsule (each of which includes a sustained release or timed release formulation), pill, powder, or granule).
  • Another exemplary such pharmaceutical composition is a liquid dosage form suitable for oral administration (e.g., an elixir, tincture, suspension, syrup, solution or emulsion).
  • the compounds of the invention may also be administered in intravenous (bolus or infusion), intraperitoneal, subcutaneous, or intramuscular form, all using dosage forms well known to those of ordinary skill in the pharmaceutical arts.
  • the dosage regimen for the compounds of the invention will, of course, vary depending upon known factors, such as the pharmacodynamic characteristics of the particular agent and its mode and route of administration; the species, age, sex, health, medical condition, and weight of the recipient; the nature and extent of the symptoms; the kind of concurrent treatment; the frequency of treatment; the route of administration, the renal and hepatic function of the patient, and the effect desired.
  • a physician can determine and prescribe the effective amount of the drug required to prevent, counter, or arrest the progress of the disease.
  • the daily oral dosage of each active ingredient when used for the indicated effects, will range between about 0.001 to 1000 mg/kg of body weight, preferably between about 0.01 to 100 mg/kg of body weight per day, and most preferably between about 1 .0 to 20 mg/kg/day.
  • the most preferred doses will range from about 1 to about 10 mg/kg/minute during a constant rate infusion.
  • Compounds of the invention may be administered in a single daily dose, or the total daily dosage may be administered in divided doses of two, three, or four times daily.
  • Compounds of the invention can be administered in intranasal form via topical use of suitable intranasal vehicles, or via transdermal routes, using transdermal skin patches.
  • suitable intranasal vehicles or via transdermal routes, using transdermal skin patches.
  • the dosage administration will, of course, be continuous rather than intermittent throughout the dosage regimen.
  • the compounds are typically administered alongside suitable pharmaceutical diluents, excipients, or carriers (collectively referred to herein as pharmaceutical carriers) suitably selected with respect to the intended form of administration, that is, oral tablets, capsules, elixirs, syrups and the like, and consistent with conventional pharmaceutical practices.
  • suitable pharmaceutical diluents, excipients, or carriers suitably selected with respect to the intended form of administration, that is, oral tablets, capsules, elixirs, syrups and the like, and consistent with conventional pharmaceutical practices.
  • the active drug component can be combined with an oral, non-toxic, pharmaceutically acceptable, inert carrier such as lactose, starch, sucrose, glucose, methyl cellulose, magnesium stearate, dicalcium phosphate, calcium sulfate, mannitol, sorbitol and the like; for oral administration in liquid form, the oral drug components can be combined with any oral, non-toxic, pharmaceutically acceptable inert carrier such as ethanol, glycerol, water, and the like.
  • suitable binders, lubricants, disintegrating agents, and colouring agents can also be incorporated into the mixture.
  • Suitable binders include starch, gelatin, natural sugars such as glucose or beta-lactose, com sweeteners, natural and synthetic gums such as acacia, tragacanth, or sodium alginate, carboxymethylcellulose, polyethylene glycol, waxes, and the like.
  • Lubricants used in these dosage forms include sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride, and the like.
  • Disintegrators include, without limitation, starch, methyl cellulose, agar, bentonite, xanthan gum, and the like.
  • Dosage forms suitable for administration may contain from about 1 milligram to about 100 milligrams of active ingredient per dosage unit.
  • the active ingredient will ordinarily be present in an amount of about 0.5-95% by weight based on the total weight of the composition.
  • Suitable pharmaceutical carriers are described in Remington's Pharmaceutical Sciences, Mack Publishing Company, a standard reference text in this field.
  • the compounds 1 can be prepared by sequential conversion to the amides 5 by treatment with hydroxiadamantylamine and a coupling agent (eg, HATU) and subsequent Suzuki coupling with a boronic acid or ester in the presence of a palladium catalyst like [1 ,1 '-Bis(diphenylphosphino) ferrocene] palladium(ll) dichloride.
  • a coupling agent eg, HATU
  • a boronic acid or ester in the presence of a palladium catalyst like [1 ,1 '-Bis(diphenylphosphino) ferrocene] palladium(ll) dichloride.
  • a palladium catalyst like [1 ,1 '-Bis(diphenylphosphino) ferrocene] palladium(ll) dichloride.
  • compounds 1 can be prepared by amidation of carboxylic acids 6 using the aforementioned conditions.
  • Reaction products were purified, when necessary, by flash chromatography on silica gel (40- 63 pm) with the solvent system indicated. Purifications in reverse phase were made in a Biotage Isolera® automated purification system equipped with a C18 column and using a gradient, unless otherwise stated, of water-acetonitrile/MeOH (1 :1 ) (0.1 % v/v ammonium formate both phases) from 0% to 100% acetonitrile/MeOH (1 :1 ) in 40 column volumes. The appropriate fractions were collected and the solvents evaporated under reduced pressure and/or freeze dried.
  • the UPLC chromatographic separations were obtained using a Waters Acquity UPLC system coupled to a SQD mass spectrometer detector.
  • the system was equipped with an ACQUITY UPLC BEH C-18 (2.1x50 mm, 1 .7 mm) column.
  • the mobile phase was formic acid (0.4 mL), ammonia (0.1 mL), methanol (500 mL) and acetonitrile (500 mL) (B) and formic acid (0.5 mL), ammonia (0.125 mL) and water (1000 mL) (A).
  • a gradient between 0 to 95% of B was used.
  • the run time was 3 or 6 minutes.
  • the injection volume was 0.5 microliter. Chromatograms were processed at 210 nM or 254 nM. Mass spectra of the chromatograms were acquired using positive and negative electrospray ionization.
  • Pd2(dba)3 tris(dibenzylidineacetone)dipalladium(0)
  • DIPEA Diisopropylethylamine
  • HATU 1-(bis(dimethylamino)methylene)-1 H-[1 ,2,3]triazolo[4,5-b]pyridine-1-ium 3-oxide hexafluorophosphate(V)
  • N-bromosuccinimide (2.06 g, 11.56 mmol, 980.69 pL) was added to a solution of 6- bromopyridin-2-amine (2 g, 11.56 mmol) in Dry DMF (27.50 mL) in portions over 30 minutes at 0 °C.
  • the reaction mixture was stirred at 25 °C for 16 hr.
  • the reaction mixture was poured into water and the precipitate was filtered, washed with water, and dried to obtain 5,6- dibromopyridin-2-amine (2.59 g, 10.16mmol, 88% yield) as a white solid.
  • the reaction mixture was then cooled, diluted with Ethyl acetate and washed with water, brine, dried and concentrated on vacuo.
  • the crude was purified by silica gel column chromatography in dichloromethane ZMeOH (from 100:0 to 99.5:0.5). The product fractions were combined dichloromethane and concentrated to obtain 5-bromo-6-(3-fluorophenyl)pyridin-2-amine (304.20 mg, 1 .12mmol, 47% yield) as a yellow solid.
  • Step 4 3-bromo-6-(3-fluorophenyl)-5-(4-pyridyl)pyridin-2-amine
  • N-bromosuccinimide (163.89 mg, 0.92 mmol) was added to a solution of 6-(3-fluorophenyl)-5- (4-pyridyl)pyridin-2-amine (0.235 g, 0.88 mmol)in dry DMF (2.5 mL) in portions over 30 minutes at 0 °C.
  • the reaction mixture was stirred at 25 °C for 16 hr.
  • the reaction mixture was poured into water and was extracted with Ethyl acetate, dried and concentrated in vacuo.
  • the crude was purified by silica gel column chromatography in heptanes/Ethyl acetate (from 100:0 to 70:30).
  • Step 5 6-(3-fluorophenyl)-5-(4-pyridyl)-3-(2-trimethylsilylethynyl)pyridin-2-amine
  • Step 6 3-ethynyl-6-(3-fluorophenyl)-5-(4-pyridyl)pyridin-2-amine
  • Potassium carbonate (29.10 mg, 0.2 1 mmol) was added to a solution of 6-(3-fluorophenyl)-5- (4-pyridyl)-3-(2-trimethylsilylethynyl)pyridin-2-amine (0.233 g, 0.64 mmol) in methanol (2 mL) .
  • the reaction mixture was stirred at 25 °C for 1 hr .
  • the reaction mixture was partitioned between Ethyl acetate/H2O:brine. After phase separation, the product was extracted with Ethyl acetate. The combined organic layers were dried and concentrated.
  • 5-bromo-6-(2-furyl)pyrazin-2-amine 250 mg, 1.04 mmol
  • 4-(4, 4,5,5- tetramethyl-1 ,3,2-dioxaborolan-2-yl)pyridine 426.5 mg, 2.08 mmol
  • 2M caesium carbonate 3.12 mL, 6.24 mmol
  • Pd(dppf)CI2 ⁇ CH2CI2 39.6 mg, 0.05 mmol
  • 3-bromo-6-(2-furyl)-5-(4-pyridyl)pyrazin-2-amine 90 mg, 0.28 mmol
  • ethynyl(trimethyl)silane 80 uL, 0.56 mmol
  • PdCI2(PPh3)2 8 mg, 0.01 mmol
  • iodocopper 2.2 mg, 0.01 mmol
  • Step 6 3-ethynyl-6-(furan-2-yl)-5-(pyridin-4-yl)pyrazin-2-amine
  • N-bromosuccinimide (2.06 g, 11.56 mmol, 980.69 pL) was added to a solution of 6- bromopyridin-2-amine (2 g, 11.56 mmol) in Dry DMF (27.50 mL) in portions over 30 minutes at 0 °C.
  • the reaction mixture was stirred at 25 °C for 16 hr.
  • the reaction mixture was poured into water and the precipitate was filtered, washed with water, and dried to obtain 5,6- dibromopyridin-2-amine (2.59 g, 10.16mmol, 88% yield) as a white solid.
  • Step 4 3-bromo-5-(2,6-dimethyl-4-pyridyl)-6-(3-fluorophenyl)pyridin-2-amine
  • N-bromosuccinimide (126.14 mg, 0.71 mmol) was added to a solution of 5-(2,6-dimethyl-4- pyridyl)-6-(3-fluorophenyl)pyridin-2-amine (0.200 g, 0.67 mmol) in dry DMF (2.6 mL) in portions over 30 minutes at 0 °C.
  • the reaction mixture was stirred at 25 °C for 4 hr, The reaction mixture was poured into water and was extracted with Ethyl acetate, dried over anh. MgSO4, filtered and concentrated in vacuo.
  • the crude was purified by silica gel column chromatography in Heptanes/Ethyl acetate (from 100:0 to 30:70).
  • Step 5 5-(2,6-dimethyl-4-pyridyl)-6-(3-fluorophenyl)-3-(2-trimethylsilylethynyl)pyridin-2- amine
  • Step 6 5-(2,6-dimethyl-4-pyridyl)-3-ethynyl-6-(3-fluorophenyl)pyridin-2-amine
  • the crude was purified by flash column chromatography on silica eluting with heptantes/Ethyl acetate from 100:0 to 50:50.
  • the product fractions were combined and concentrated to yield 5-(2,6-dimethyl-4-pyridyl)-3-ethynyl-6-(3-fluorophenyl)pyridin-2-amine (0.0955 g, 0.30mmol, 93% yield) as a black solid.
  • Step 1 6-(3-fluorophenyl)pyrazin-2-amine
  • 5-bromo-6-(3-fluorophenyl)pyrazin-2-amine 300 mg, 1.11 mmol
  • 4-(4, 4,5,5- tetramethyl-1 ,3,2-dioxaborolan-2-yl)pyridine 459 mg, 2.23 mmol
  • 2M caesium carbonate 3.3 mL, 6.66 mmol
  • Pd(dppf)CI2 ⁇ CH2CI2 45 mg, 0.055 mmol
  • reaction mixture was then cooled, diluted with Ethyl acetate and washed with water, brine, dried and concentrated on vacuo.
  • the crude was purified by silica gel column chromatography in dichloromethane /MeOH (from 2% to 5%). The product fractions were combined and concentrated to obtain 6-(3-fluorophenyl)-5-(pyridin-4-yl)pyrazin-2-amine (0.178 g, 60% yield) as a yellowish solid.
  • Tetrakis(triphenylphosphine)-palladium(0) (102.18 mg, 0.09 mmol) was added and the reaction mixture was purged with N2 for 2 min, closed and heated at 110 °C for 16 hr. The reaction mixture was then cooled, diluted with Ethyl acetate and washed with water, brine, dried over anh. MgSO4, filtered and concentrated in vacuo. The crude was purified by silica gel column chromatography in dichloromethane /MeOH (from 100:0 to 99.5:0.5). The product fractions were combined and concentrated to obtain 5-bromo-6-phenyl-pyridin-2-amine (0.4214 g, 1 .66mmol, 56% yield) as a yellow oil.
  • the reaction mixture was then cooled, diluted with Ethyl acetate and washed with water, brine, dried and concentrated in vacuo.
  • the crude was purified by silica gel column chromatography in dichloromethane /MeOH (from 100:0 to 96:4).
  • the product fractions were combined and concentrated to obtain 6-phenyl-5- (3-pyridyl)pyridin-2-amine (0.326 g, 1.31 mmol, 86% yield) as a beige solid.
  • N-bromosuccinimide (222.95 mg, 1.25 mmol) was added to a solution of 6-phenyl-5-(3- pyridyl)pyridin-2-amine (0.298 g, 1.19 mmol) in Dry DMF (3.6 mL) in portions over 30 minutes at 0 °C. The reaction mixture was stirred at 25 °C for 3 hr. The reaction mixture was poured into water and was extracted with Ethyl acetate, dried and concentrated in vacuo. The crude was purified by silica gel column chromatography in heptanes/Ethyl acetate (from 100:0 to 68:32). The product fractions were combined and concentrated to obtain3-bromo-6-phenyl-5- (3-pyridyl)pyridin-2-amine (0.284 g, 0.86mmol, 72% yield) as a pink solid.
  • 3-bromo-6-phenyl-5-(3-pyridyl)pyridin-2-amine (0.280 g, 0.85 mmol)
  • ethynyl(trimethyl)silane 500.80 mg, 5.10mmol, 720.58 pL
  • N,N-diethylethanamine (343.97 mg, 3.40 mmol, 473.79 pL) were dissolved in dry DMF (5.55 mL) and the mixture was purged with N2 for 5 min.
  • 3,5- difluorophenyl)boronic acid 155.15 mg, 0.98 mmol
  • sodium carbonate 208.27 mg, 1.97 mmol
  • tetrakis(triphenylphosphine)-palladium (0) 45.41 mg, 0.04 mmol
  • the reaction mixture was purged with N2 for 2 min, closed and heated at 110 °C for 24 hr.
  • the reaction mixture was filtered over a pad of Celite® and rinsed with Ethyl acetate. The filtrate was washed with water/brine and the water layer was extracted with Ethyl acetate. The organic layer was dried and concentrated.
  • N-bromosuccinimide 14.56 mg, 0.08 mmol was added to a stirred solution of 6-(3,5- difluorophenyl)-5-(3-pyridyl)pyridin-2-amine (215 mg, 0.75 mmol) in dry dioxane (1.94 mL) at 0°C. Then the reaction mixture was stirred and heated at 25 °C for 1 hr. The reaction mixture was diluted with Ethyl acetate and washed with brine. The organic layer was dried, filtered and concentrated. The crude was purified by flash column chromatography on silica gel eluting with heptanes/Ethyl acetate from 100:0 to 64:36.
  • 3-bromo-6-(3,5-difluorophenyl)-5-(3-pyridyl)pyridin-2-amine (205 mg, 0.56 mmol), ethynyl(trimethyl)silane (660.46 mg, 6.72 mmol, 950.30 pL), N,N-diethylethanamine (453.63 mg, 4.48 mmol, 624.84 pL) were dissolved in dry DMF (1 .68 mL) and the mixture was purged with N2 for 5 min.
  • the reaction mixture was filtered over a pad of Celite® and rinsed with Ethyl acetate. The filtrate was washed with water/brine and the water layer was extracted with Ethyl acetate. The organic layer was dried and concentrated.
  • the crude was purified by flash column chromatography on silica gel eluting with heptanes/Ethyl acetate from 100:0 to 80:20. The product fractions were combined and concentrated to yield5-bromo-6-(3-fluorophenyl)pyridin-2-amine (378 mg, 1.39mmol, 71 % yield) as an ivory solid.
  • the reaction mixture was recharged with 5-bromo-6-(3-fluorophenyl)pyridin-2-amine (0.378 g, 1.40mmol), 3- pyridylboronic acid (344.43 mg, 2.80 mmol) and caesium carbonate (456.49 mg, 1.40 mmol). Then, Pd(dppf)CI2 ⁇ CH2CI2 (11.44 mg, 0.01 mmol) was added and the reaction mixture was purged with N2 for 2 min, closed and heated at 110 °C for 16 hr. The reaction mixture was filtered over a pad of Celite® and rinsed with Ethyl acetate.
  • N-bromosuccinimide 14.56 mg, 0.08 mmol was added to a stirred solution of 6-(3- fluorophenyl)-5-(3-pyridyl)pyridin-2-amine (0.209 g, 0.73 mmol) in dry dioxane (2.5 mL) at 0°C . Then the reaction mixture was stirred and heated at 25 °C for 16 hr. The reaction mixture was diluted with Ethyl acetate and was washed with brine. The organic layer was dried and concentrated. The crude was purified by flash column chromatography on silica gel eluting with Heptanes/Ethyl acetate from 100:0 to 50:50.
  • Step 4 6-(3-fluorophenyl)-5-(3-pyridyl)-3-(2-trimethylsilylethynyl)pyridin-2-amine
  • Potassium carbonate (21.42 mg, 0.15 mmol, 9.35 pL) was added to a solution of 6-(3- fluorophenyl)-5-(3-pyridyl)-3-(2-trimethylsilylethynyl)pyridin-2-amine (0.175 g, 0.47 mmol) in MeOH (1 .5 mL).
  • the reaction mixture was stirred at 25 °C for 1 hr.
  • the reaction mixture was partitioned between Ethyl acetate/H2O:brine. After phase separation, the product was extracted with Ethyl acetate. The combined organics layers were dried and concentrated.
  • 3-pyridylboronic acid (265.69 mg, 2.16 mmol), caesium carbonate (640.24 mg, 1.97mmol) and Pd(dppf)CI2-CH2CI2 (16.05 mg, 0.02 mmol) were added and the reaction mixture was purged with N2 for 2 min, closed and heated at 110 °C for 16 hr.
  • 3-pyridylboronic acid (169.07 mg, 1.38mmol) and caesium carbonate (640.24 mg, 1.97 mmol) and Pd(dppf)CI2-CH2CI2 (48.14 mg, 0.06mmol) were still added and the reaction mixture was purged with N2 for 2 min, closed and heated at 110 °Cf or 16 hr.
  • the reaction mixture was filtered over a pad of Celite® and rinsed with Ethyl acetate. The filtrate was washed with water/brine and the water layer was extracted with Ethyl acetate. The organic layer was dried and concentrated.
  • the crude was purified by silica gel column chromatography in dichloromethane /MeOH (from 100:0 to 94:6). The product fractions were combined and concentrated to obtain 5,6-bis(3-pyridyl)pyridin-2-amine (0.257 g, 1.02mmol, 52% yield) as a grey solid.
  • N-bromosuccinimide 14.56 mg, 0.08 mmol was added to a stirred solution of 5,6-bis(3- pyridyl)pyridin-2-amine (0.257 g, 1 .02 mmol) in dry dioxane (3.1 mL) at 0°C . Then the reaction mixture was stirred and heated at 25 °C for 16 hr. The reaction mixture was diluted with Ethyl acetate and washed with brine. The organic layer was dried and concentrated. The crude was purified by flash column chromatography on silica gel eluting with dichloromethane /MeOH from 100:0 to 98:2. The product fractions were combined and concentrated to yield 3-bromo- 5,6-bis(3-pyridyl)pyridin-2-amine (0.2363 g, 0.71 mmol, 69% yield) as a beige solid.
  • Keratinocytes were isolated from surplus skin samples sourced for diagnostic purposes after obtaining patient informed consent and Ethical Committee approval (reference HULP: PI- 3911 , Hospital Universitario de la Paz, Madrid, Spain). These keratinocytes were used to generate immortalized cell lines. Immortalization resulted from the forced expression of human papillomavirus HPV16 E6/E7 antigens (Chamorro et al 2013). Skin keratinocytes were isolated as previously described (Chamorro et al 2013).
  • retroviral particles obtained from the supernatant of HEK293T cells transfected with pLXSN16E6E7, pNG-VL3- MLV-gag-pol and pNGVL3-4070 plasmids (Halbert et al 1991 , Yang et al 1999, (Chamorro et al 2013).
  • the cells were kept in KGM medium in the presence of feeder layer and were amplified during several passages at a ratio of 1 :4 in each passage. After 18 passages, the keratinocytes were cultured in the absence of feeder layer in KGM medium.
  • RDEB_L1 In addition to the mutation of RDEB_L1 , one mutation previously shown to respond well to aminoglycosides (Cogan et al 2014) was recombinantly generated and stably transfected into HEK293 cells, a cell line that does not express C7, to generate HEK293R 578X .
  • site directed mutagenesis was used to generate a mutated Col7A1 cDNA containing a C to T change at nucleotide 1732, resulting in a change in Arg578 to an UAG termination codon.
  • the resulting mutated COL7A1 cDNA was then stably transfected into HEK293 cells after verifying the absence of expression of COL7A1 in this cell line. Zeocin was used as selection antibiotic to obtain the stably transfected clones which were selected by limiting dilution.
  • RDEB_L1 and HEK293 R578X cell lines were characterized using western blot and an antibodybased AlphaLISA assay recognizing the N-terminal part of C7 (amino acids 36-153) within the non-collagenous 1 domain (NC1 ), spanning amino acids 17-1253 (Christiano et al 1994).
  • the AlphaLISA assay was developed in house by using the biotin-conjugated rabbit polyclonal antibody anti-Collagen VII (LS-298147, LSBio) and conjugating the acceptor beads (6772001 , PerkinElmer) to a monoclonal anti-Collagen type VII antibody produced in mouse, clone LH7.2 (MA5-13432, Thermo). Formulation of the mAb clone LH7.2 was exchanged to PBS and concentrated 5x by using Amicon® Ultra-0,5 centrifugal filters up to rise 1 mg/ml. The conjugation procedure was carried out according to the protocol provided by PerkinElmer.
  • RDEB patient-derived cells and NHEK were grown in complete KGMTM GoldTM (KBMTM GoldTM Basal Medium with SingleQuotsTM) excluding the GA-100, (H3192060, Cultek).
  • KGMTM GoldTM KBMTM GoldTM Basal Medium with SingleQuotsTM
  • GA-100 GA-100
  • TripLE express solution (12604054, Gibco) was added to detach the cells.
  • Trypsin Neutralizing Solution (CC- 5002, Lonza) was added and then the cell suspension was centrifuged at 140 g for 5 minutes at room temperature, the supernatant was discarded, and the pellet was resuspended in KGM complete media, without GA-100 at desired density.
  • HEK293 5R578X were cultured in DMEM high glucose supplemented with 10% FBS and 2mM Glutamax (61965026, Thermo Fisher). PBS was used for the rinse step and TripLE express were used to detach the cells.
  • DMEM complete media was used for neutralizing the TripLE express and after centrifugation at 140 g for 5 minutes at room temperature, the supernatant was discarded, and cells were resuspended in DMEM complete media at working densities.
  • 5,000 cells were plated in 20pl of supplemented KGM media containing 25ng/ml TGFP2 (GF113, SIGMA) without GA-1000 in 384-well cell culture plates (781098, Greiner). The day after, 5pl of test compounds prepared 5x in supplemented KGM without GA-1000 were added over cells and incubated for 48h at 37°C and 5% CO2.
  • HEK293 R578X 10,000 cells were plated in 20pl of DMEM (High Glucose, 2mM L-Glutamine, sodium pyruvate and FBS 10%) in 384-well cell culture plates (781098, Greiner). The day after, 5pl of test compounds prepared 5x in DMEM complete medium were added over cells and incubated for 48h at 37°C and 5% CO2.
  • the viability of RDEB or NHEK was evaluated by measuring the ATP levels as indirect read out of the potential cytotoxicity effect of compounds.
  • Cells were plated and treated exactly with the same conditions followed in the AlphaLISA assay, described above.
  • For measuring the ATP levels cells were processed using the ATPIiteTM 1 step luminescence-based assay (PerkinElmer 6016731 ), following the instructions provided by the supplier. 25 pl of ATPIiteTM reagent was added directly over cells and plates were covered with a sealing film and shaken for 25 min using a microtiter plate shaker. Luminescence was measured in Luminoskan Ascent luminometer (Thermo Fisher Scientific). 10pM of Staurosporine was used as reference compound and control for the total cytotoxicity.
  • RDEB keratinocytes or normal human epidermal keratinocytes were plated in 96-well High Content Imaging microplate (Corning 4517) at 10,000 cells per well in 80pl of supplemented KGM media without GA-1000, media used for RDEB also was supplemented with 25ng/ml of TGF02 (TGF0), and cells were incubated at 37°C and 5%CO2. The day after, 20 pl of test compounds prepared 5x in supplemented KGM without GA-100 were added and incubated for 48h at 37°C and 5%CO2. Cells were permeabilized aspirating the media and adding 50pl of 100% ice-cold methanol for 15 minutes at 4°C.
  • blocking buffer PBS, 5% serum goat and 0.3% Triton X-100
  • Blocking buffer was aspirated and 50ul of 3pg/ml of primary antibody anti-collagen type VII (HPA042420, SIGMA) diluted in diluent buffer (PBS, 1 % BSA and 0.3% Triton X-100) was added and incubated for 2h at room temperature.
  • Collagen VII was monitored analyzing the total region intensity red and cell counting was monitored by object counting in the DAPI channel.
  • 15,000 RDEB keratinocytes or normal human epidermal keratinocytes were plated in 96-well cell culture plate in 80 pl of supplemented KGM media without GA-1000, and containing 25ng/ml of TGF02 for RDEB keratinocytes, and cells were incubated overnight at 37°C and 5%CO2. The day after, cells were treated with 20 l of test compounds prepared 5x in supplemented KGM without GA-100, for 24h at 37°C and 5%CO2.
  • RNA extraction was conducted using the RNeasy 96 Kit (4) (74181 , Qiagen) and following the instructions provided by QIAGEN. Total RNA was recovered in 50 pL of RNase-free water and the yield of extracted RNA was quantified using a NanoDrop 2000 Spectrophotometer (ThermoFisher). Purified RNA was stored at -80°C until RT-qPCR. Real time-qPCR was performed by using the iTaq Universal Probes One-Step Kit, (1725140, BioRad) and according to the manufacturer’s instructions.
  • RTqPCR were run in QuantStudioTM Real-Time PCR System (Applied Biosystems) or in CFX96 Touch Real-Time PCR Detection System (BioRad) for 384 or 96 well format respectively.
  • COL7A1 Hs00164310_m1
  • GAPDH Hs02758991_g1
  • TaqManTM probe Thermo Fisher Scientific
  • RT-qPCR was performed to quantify the COL7A1 mRNA (exon 64) expression.
  • human GAPDH was used as the endogenous control. Results were analyzed with the QuantStudio Real-Time PCR Software.
  • Patient derived RDEB_L1 transformed keratinocytes and the recombinant HEK293 R578 cell line were generated as described in the methodology section and characterized using AlphaLISA and western blot.
  • Patient-derived transformed cell line RDEB_L1 , containing a bi- allelic PTC mutation in exon 72 was selected based on its response to aminoglycosides G418 and gentamicin.
  • G418 showed more potent activity than gentamicin with an EC50 around 3 orders of magnitude lower than gentamicin (Figs. 1 a, 1 b).
  • gentamicin upregulated full length and truncated C7 in RDEB_L1 and HEK293 R578X cells (Figs 1 c, 1d).
  • Gentamicin showed upregulation of full length C7 and truncated C7 at concentrations of 2.09 and 4.18 mM in RDEB_L1 (Fig 5c) and in HEK293R 578X cells at a concentration of 4.18 mM (Fig 1 c).
  • the size of the truncated C7 products detected in the western blot was compatible with their predicted molecular weights, based on the location of the PTC mutations (Fig 1c, d): 202 kD (RDEB_L1 ) and 62 kD (HEK293 R578X ).
  • a library of compounds was evaluated at 25 pM in RDEB_L1 cells, a concentration intended to capture hits with a potency comparable to the relatively potent reference aminoglycoside molecule G418 (EC50 4.9 pM) and more potent than the reference drug gentamicin (EC50 1.7 mM) (Table 1 ).
  • Detection of total C7 protein using AlphaLISA was used as a gain of signal readout. This had the advantage of minimizing the possibility to identify hits with cytotoxic properties, as cytotoxicity or reduced viability would be expected to have an impact in the cell translational machinery, precluding the expression of C7. Following a primary screen, the activity of the hits was confirmed again by AlphaLISA and their potential interference was verified testing their activity in the same assay in the absence of cells.
  • the compounds in Table 1 showed an increase in C7 levels in RDEB_L1 cells.
  • Relative EC50 and Emax are shown as arithmetic mean ⁇ standard deviation (SD).
  • SD standard deviation
  • Viability was measured using ATP levels in normal human keratinocytes (NHK), RDEB_L1 patient-derived cells and the HEK293 R578X recombinant cell line.
  • EXAMPLE 7 did not induce non-specific readthrough above background levels under conditions where gentamicin showed non-specific readthrough activity, at concentrations 2 fold (gentamicin) and 3 fold (EXAMPLE 7) over their C7 AlphaLISA EC50 in RDEB_L1 cells ( Figure 6).

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Abstract

La présente invention concerne des composés de pyrazine ou de pyridine induisant l'expression du collagène VII (C7). La présente invention concerne également des compositions pharmaceutiques les contenant, des procédés pour leur préparation et leur utilisation dans le traitement de plusieurs troubles.
PCT/EP2024/072592 2023-08-11 2024-08-09 Dérivés de pyridine ou de pyrazine utilisés en tant qu'inducteurs de collagène vii (c7) et leur utilisation en thérapie Pending WO2025036840A1 (fr)

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Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20040180905A1 (en) * 2003-03-11 2004-09-16 Pfizer Inc Novel pyrazine compounds as transforming growth factor (TGF) compounds
WO2007017096A1 (fr) * 2005-07-29 2007-02-15 Laboratorios Almirall, S.A. Derives de pyrazine utiles en tant qu'antagonistes de recepteur d'adenosine
WO2019018584A1 (fr) * 2017-07-18 2019-01-24 GiraFpharma LLC Composés hétérocycliques utilisés en tant qu'antagonistes de l'adénosine

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20040180905A1 (en) * 2003-03-11 2004-09-16 Pfizer Inc Novel pyrazine compounds as transforming growth factor (TGF) compounds
WO2007017096A1 (fr) * 2005-07-29 2007-02-15 Laboratorios Almirall, S.A. Derives de pyrazine utiles en tant qu'antagonistes de recepteur d'adenosine
WO2019018584A1 (fr) * 2017-07-18 2019-01-24 GiraFpharma LLC Composés hétérocycliques utilisés en tant qu'antagonistes de l'adénosine

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"Remington's Pharmaceutical Sciences", MACK PUBLISHING COMPANY
AGHAMOHAMMADZADEH S.: "1122 ZKN-0013 promotes premature termination codon readthrough to restore collagen VII protein expression and function in recessive dystrophic epidermolysis bullosa (RDEB)", JOURNAL OF INVESTIGATIVE DERMATOLOGY, 17 April 2023 (2023-04-17), pages S192, XP093217441, Retrieved from the Internet <URL:https://www.sciencedirect.com/science/article/pii/S0022202X2301309X?via%3Dihub> *
FORNS PILAR ET AL: "Pyrazine-based Syk kinase inhibitors", BIOORGANIC & MEDICINAL CHEMISTRY LETTERS, vol. 22, no. 8, 1 April 2012 (2012-04-01), Amsterdam NL, pages 2784 - 2788, XP093091878, ISSN: 0960-894X, DOI: 10.1016/j.bmcl.2012.02.087 *
JOVER IRENE ET AL: "Identification of novel small molecule-based strategies of COL7A1 upregulation and readthrough activity for the treatment of recessive dystrophic epidermolysis bullosa", SCIENTIFIC REPORTS, vol. 14, no. 1, 16 August 2024 (2024-08-16), US, XP093217299, ISSN: 2045-2322, Retrieved from the Internet <URL:https://www.nature.com/articles/s41598-024-67398-8> DOI: 10.1038/s41598-024-67398-8 *
ORELLANA ADELINA ET AL: "Application of a phenotypic drug discovery strategy to identify biological and chemical starting points for inhibition of TSLP production in lung epithelial cells", PLOS ONE, vol. 13, no. 1, 10 January 2018 (2018-01-10), US, pages e0189247, XP093217425, ISSN: 1932-6203, Retrieved from the Internet <URL:https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0189247&type=printable> DOI: 10.1371/journal.pone.0189247 *
OSIPOWICZ KATARZYNA ET AL: "Efficacy of gentamicin 0.3% solution of oral erosions healing in patients with severe generalized recessive dystrophic epidermolysis bullosa and its impact on the expression of type VII collagen", POSTEPY DERMATOLOGII I ALERGOLOGII = ADVANCES IN DERMATOLOGY AND ALLERGOLOGY, vol. 38, no. 6, 1 January 2021 (2021-01-01), POLAND, pages 979 - 984, XP093217336, ISSN: 1642-395X, Retrieved from the Internet <URL:https://pmc.ncbi.nlm.nih.gov/articles/PMC8802963/pdf/PDIA-38-41227.pdf> DOI: 10.5114/ada.2020.97072 *
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