WO2017037567A1

WO2017037567A1 - Regulators of frataxin

Info

Publication number: WO2017037567A1
Application number: PCT/IB2016/055013
Authority: WO
Inventors: Eugene Lvovich Piatnitski Chekler; Leslie Anthony DAKIN; Lyn Howard Jones; Joseph Fouad NABHAN; Mathew Tyler PLETCHER
Original assignee: Pfizer Inc.
Priority date: 2015-09-03
Filing date: 2016-08-22
Publication date: 2017-03-09
Also published as: CA2940742A1

Abstract

This invention relates to a method of treating a condition or a disease associated with decreased levels or activity of frataxin, including Friedreich's ataxia, comprising administering to a subject in needthereof a therapeutically effective amount of a BET- family bromodomain inhibitor, or a pharmaceutically acceptable salt thereof. Related methods, pharmaceutical uses and pharmaceutical compositions are disclosed herein.

Description

REGULATORS OF FRATAXIN

FIELD

The present invention relates to regulators of frataxin and, in particular, to methods of regulating the level of frataxin or the expression of frataxin in a cell or subject, comprising administering to the cell or subject a BET-family bromodomain inhibitor, or a pharmaceutically acceptable salt thereof. Associated methods of treatment, pharmaceutical uses and pharmaceutical compositions are also described.

BACKGROUND Friedreich's ataxia (also called FA or FRDA) is a rare recessive inherited disease characterized by progressive damage to the nervous system and movement problems. The first symptoms of the condition are usually gait ataxia or difficultly walking, but overtime the symptoms worsen and spread resulting in degeneration of the spinal cord, peripheral nerves and cerebellum. The disorder also causes problems in the spine and loss of tendon reflex. Most people with Friedreich's ataxia develop scoliosis (a curving of the spine to one side), which often requires surgical intervention for treatment. Dysarthria (slowness and slurring of speech) may also develop as well as hearing and vision loss. Other symptoms that may occur include chest pain, shortness of breath, and heart palpitations. These symptoms are the result of various forms of heart disease that often accompany Friedreich's ataxia, such as hypertrophic cardiomyopathy (enlargement of the heart), myocardial fibrosis (formation of fiber-like material in the muscles of the heart), and cardiac failure. Heart rhythm abnormalities such as tachycardia (fast heart rate) and heart block (impaired conduction of cardiac impulses within the heart) are also common. About 20 percent of people with Friedreich's ataxia develop carbohydrate intolerance and 10 percent develop diabetes. Symptoms typically begin between the ages of 5 and 15 years and generally within 10 to 20 years after the appearance of the first symptoms the patient is confined to a wheelchair and in later stages of the disease individuals may become completely incapacitated. Friedreich's ataxia can lead to early death, often as a result of associated heart disease.

Friedreich's ataxia is caused by reduced expression of frataxin (FXN) as a result of inheriting two copies of the mutated FXN gene which disrupts the normal production of frataxin. Frataxin is found in the mitochondria and research suggests that without a normal level of frataxin, certain cells in the body (especially peripheral nerve, spinal cord, brain and heart muscle cells) cannot effectively produce energy and it is further hypothesized that these cells experience oxidative stress due to the buildup of toxic byproducts. Frataxin is initially synthesized in vivo as a precursor polypeptide which is in turn directed to the mitochondrial matrix where it is proteolytically cleaved by mitochondrial processing peptidase to the mature form of frataxin via a processing intermediate frataxin ("The in vitro mitochondrial two-step maturation of human frataxin." Stephane Schmucker et al, Human Molecular Genetics, 2008, vol 17 no 22, p3521- 3531). It is also thought that reduced expression of frataxin results in the nervous system, heart, and pancreas being particularly susceptible to damage from free radicals.

As with many degenerative diseases of the nervous system, there is currently no known cure or effective treatment for Friedreich's ataxia. To date, the only therapy options remain symptomatic treatment to help individuals maintain optimal functioning for as long as possible. For example, prescription of medication to treat diabetes or heart problems, use of surgery or braces or physical therapy to treat scoliosis and other skeletal problems.

There remains a need for new small molecule approaches for the treatment of Friedreich's ataxia or related conditions, diseases or disorders associated with the regulation of frataxin.

SUMMARY

This invention relates to a method of regulating the expression of frataxin in a cell or a subject, comprising administering to the cell or the subject in need thereof a therapeutically effective amount of a BET- family bromodomain inhibitor, or a pharmaceutically acceptable salt thereof.

This invention also relates to a method of increasing the expression of frataxin in a cell or a subject, comprising administering to the cell or the subject in need thereof a therapeutically effective amount of a BET- family bromodomain inhibitor, or a pharmaceutically acceptable salt thereof.

This invention also relates to a method of increasing the expression of frataxin in a cell or a subject which cell or subject has decreased frataxin expression when compared to a healthy control cell or healthy subject, comprising administering to the cell or the subject in need thereof a therapeutically effective amount of a BET- family bromodomain inhibitor, or a pharmaceutically acceptable salt thereof.

This invention also relates to a method of increasing the level of frataxin in a cell or a subject, comprising administering to the cell or the subject in need thereof a therapeutically effective amount of a BET- family bromodomain inhibitor, or a pharmaceutically acceptable salt thereof.

This invention also relates to a method of increasing the level of frataxin in a cell or a subject which cell or subject has decreased level of frataxin when compared to a healthy control cell or healthy subject, comprising administering to the cell or the subject in need thereof a therapeutically effective amount of a BET- family bromodomain inhibitor, or a pharmaceutically acceptable salt thereof.

This invention also relates to a method of treating a condition, disease or disorder, or the symptoms associated with the condition, disease or disorder, which condition, disease or disorder is associated with decreased frataxin expression, comprising administering to a subject in need thereof a therapeutically effective amount of a BET- family bromodomain inhibitor, or a pharmaceutically acceptable salt thereof.

This invention also relates to a method of treating a condition, disease or disorder, or the symptoms associated with the condition, disease or disorder, which condition, disease or disorder is associated with a decreased level of frataxin, comprising administering to a subject in need thereof a therapeutically effective amount of a BET- family bromodomain inhibitor, or a pharmaceutically acceptable salt thereof.

This invention also relates to a method of treating Friedreich's ataxia comprising administering to a subject in need thereof a therapeutically effective amount of a BET- family bromodomain inhibitor, or a pharmaceutically acceptable salt thereof.

This invention also relates to a BET-family bromodomain inhibitor, or a pharmaceutically acceptable salt thereof, for use in the treatment of a condition, disease or disorder, or the symptoms associated with the condition, disease or disorder, which condition, disease or disorder is associated with decreased frataxin expression.

This invention also relates to a BET-family bromodomain inhibitor, or a pharmaceutically acceptable salt thereof, for use in the treatment of a condition, disease or disorder, or the symptoms associated with the condition, disease or disorder, which condition, disease or disorder is associated with a decreased level of frataxin. This invention also relates to a BET-family bromodomain inhibitor, or a pharmaceutically acceptable salt thereof, for use in the treatment of Friedreich's ataxia.

This invention also relates to the use of a BET-family bromodomain inhibitor, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for the treatment of a condition, disease or disorder, or the symptoms associated with the condition, disease or disorder, which condition, disease or disorder is associated with decreased frataxin expression.

This invention also relates to the use of a BET-family bromodomain inhibitor, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for the treatment of a condition, disease or disorder, or the symptoms associated with the condition, disease or disorder, which condition, disease or disorder is associated with a decreased level of frataxin.

This invention also relates to the use of a BET-family bromodomain inhibitor, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for the treatment of Friedreich's ataxia.

This invention also relates to a pharmaceutical composition comprising a BET- family bromodomain inhibitor, or a pharmaceutically acceptable salt thereof, for the treatment of a condition, disease or disorder, or the symptoms associated with the condition, disease or disorder, which condition, disease or disorder is associated with decreased frataxin expression.

This invention also relates to a pharmaceutical composition comprising a BET- family bromodomain inhibitor, or a pharmaceutically acceptable salt thereof, for the treatment of a condition, disease or disorder, or the symptoms associated with the condition, disease or disorder, which condition, disease or disorder is associated with a decreased level of frataxin.

This invention also relates to a pharmaceutical composition comprising a BET- family bromodomain inhibitor, or a pharmaceutically acceptable salt thereof, for the treatment of Friedreich's ataxia.

BRIEF DESCRIPTION OF FIGURES

Figure 1 shows that BET-family bromodomain inhibitors IBET-762 and JQ-1 (+) increased levels of both intermediate frataxin (bands "i") and mature frataxin (bands "m") at 72 hrs by Western blot (two exposures shown) in FRDA GM15850 cells. Figure 2 shows the cytotoxic effects of treatment of the FRDA GM 15850 cells with BET-family bromodomain inhibitors IBET-762 and JQ-1 (+) (protein concentrations (mg/ml) for the resultant lysates).

Figure 3 shows that a minimally cyctotoxic concentration of BET-family bromodomain inhibitor JQ-1 (+) increased levels of both intermediate frataxin (bands "i") and mature frataxin (bands "m") at 72 hrs by Western blot (two exposures shown) in FRDA GM 15850 cells. Results from DMSO treated AG14725 healthy cells control are also shown.

Figure 4 shows pixel densitometry quantification (base line corrected) of immunopositive bands corresponding to mature frataxin from the Western blot of Figure 3. An approximately 1.5 fold increase was observed at 25nM and 50nM treatment with BET-family bromodomain inhibitor JQ-1 (+) when compared to DMSO vehicle treated controls (p<0.05).

Figure 5 shows a similar cellular toxicity profile (LD₅₀) for both FRDA GM 15850 lymphoblast cells and healthy AG14725 upon treatment with BET-family bromodomain inhibitor JQ-1 (+), suggesting that a lower level of frataxin does not sensitize cells to toxicitiy by the BET-family bromodomain inhibitor JQ-1 (+).

Figure 6 shows that BET-family bromodomain inhibitors with a variety of chemical scaffolds (compounds 1 - 6), increased levels of both intermediate frataxin (bands "i") and mature frataxin (bands "m") at 72 hrs by Western blot (two exposures shown) in FRDA GM 15850 cells.

Figure 7 shows that BRD4 knockdown using the BRD4 siRNA smartpool or treatment with BET-family bromodomain inhibitor JQ-1 (+) increased levels of both intermediate frataxin (bands "i") and mature frataxin (bands "m") at 72 hrs by Western blot (two exposures shown) in FRDA GM03665 fibroblasts.

Figure 8 shows that knockdown of other BET-family proteins using siRNA smartpools also resulted in increased levels of both intermediate frataxin (bands "i") and mature frataxin (bands "m") at 72 hrs by Western blot (two exposures shown) in FRDA GM03665 fibroblasts.

DETAILED DESCRIPTION

The present invention relates to methods of regulating the level of frataxin or the expression of frataxin in a cell or subject, comprising administering to the cell or subject a BET-family bromodomain inhibitor, or a pharmaceutically acceptable salt thereof. The present invention also relates to novel pharmaceutical methods, and associated methods of treatment, utilising a BET-family bromodomain inhibitor, or a pharmaceutically acceptable salt thereof.

Methods of Treatment

This invention also relates to a method of treating a condition, disease or disorder, or the symptoms associated with the condition, disease or disorder, which condition, disease or disorder is associated with decreased frataxin expression, comprising administering to a subject in need thereof a therapeutically effective amount of a BET- family bromodomain inhibitor, or a pharmaceutically acceptable salt thereof. This invention also relates to a method of treating a condition, disease or disorder, or the symptoms associated with the condition, disease or disorder, which condition, disease or disorder is associated with a decreased level of frataxin, comprising administering to a subject in need thereof a therapeutically effective amount of a BET- family bromodomain inhibitor, or a pharmaceutically acceptable salt thereof.

This invention also relates to a BET-family bromodomain inhibitor, or a pharmaceutically acceptable salt thereof, for use in the treatment of a condition, disease or disorder, or the symptoms associated with the condition, disease or disorder, which condition, disease or disorder is associated with a decreased level of frataxin.

This invention also relates to a BET-family bromodomain inhibitor, or a pharmaceutically acceptable salt thereof, for use in the treatment of Friedreich's ataxia.

In one embodiment, as used herein, the terms "treat" or "treating" refer to partially or completely alleviating, inhibiting, ameliorating and/or relieving a condition, disease or disorder, or the symptoms associated with the condition, disease or disorder from which a patient or subject has been diagnosed as suffering or from which a patient or subject is suspected to suffer.

In another embodiment, as used herein, the terms "treat" or "treating" also refer to slowing of the advancement of a condition, disease or disorder, or the symptoms associated with the condition, disease or disorder from which a patient or subject has been diagnosed as suffering or from which a patient or subject is suspected to suffer.

In another embodiment, the terms "treat" or "treating" also refer to regression of the condition, disease or disorder, including regression of the symptoms associated with the condition, disease or disorder, from which a patient or subject has been diagnosed as suffering or from which a patient or subject is suspected to suffer.

In another embodiment, the terms "treat" or "treating" also refer to the prevention of a condition, disease or disorder, or the symptoms associated with the condition, disease or disorder from which a patient or subject has been diagnosed as suffering or from which a patient or subject is suspected to suffer.

In another embodiment, as used herein, the terms "treat" or "treating" also refer to the prevention of the advancement of a condition, disease or disorder, or the symptoms associated with the condition, disease or disorder, from which a patient or subject has been diagnosed as suffering or from which a patient or subject is suspected to suffer. The term "therapeutically effective" as used herein refers to a therapeutic agent or an amount of therapeutic agent that elicits a desirable biological activity or effect. In one embodiment, the term "therapeutically effective amount" as used herein, refers to that amount of the therapeutic agent sufficient to result in partially or completely alleviating, inhibiting, ameliorating and/or relieving the condition, disease or disorder, or partially or completely alleviating, inhibiting, ameliorating and/or relieving the symptoms associated with the condition, disease or disorder. In another embodiment the term refers to the amount of the therapeutic agent sufficient to result in partially or completely slowing the advancement of a condition, disease or disorder, and / or slowing the advancement of one or more of the symptoms associated with a condition, disease or disorder. In another embodiment, the term refers to the amount of therapeutic agent sufficient to cause regression of the condition, disease or disorder, and / or causing regression of one or more of the symptoms associated with the condition, disease or disorder. In another embodiment, the term refers to the amount of therapeutic agent sufficient to prevent the condition, disease or disorder, and / or to prevent one or more of the symptoms associated with the condition, disease or disorder. In another embodiment, the term refers to the amount of therapeutic agent sufficient to prevent the advancement of the condition, disease or disorder, and / or preventing the advancement of one or more of the symptoms associated with the condition, disease or disorder.

As used herein, except when noted, the terms "subject" or "patient" are used interchangeably and refer to mammals such as humans and non-human primates, as well as experimental animals such as rabbits, rats, and mice. In one embodiment the subject or patient is a human. In another embodiment the subject or patient is a male human. In another embodiment the subject or patient is a female human. Accordingly, the term "subject" or "patient" as used herein means any mammalian patient or subject to which the compounds useful for the present invention can be administered.

In one embodiment of the present invention, routine screening methods are used to either identify and/or diagnose patients for treatment according to the methods of the invention; to identify the existence of a specific condition, disease or disorder which is associated with decreased expression of frataxin; to identify the existence of a specific condition, disease or disorder which is associated with a decreased level of frataxin; to determine risk factors associated with a targeted or suspected condition, disease or disorder; or to determine the status of an existing condition, disease or disorder in the subject. These routine screening methods include, but are not limited to, for example, conventional work-ups to determine risk factors that may be associated with the targeted or suspected condition, disease or disorder; conventional genetic screening methods and the like. These and other routine methods allow the clinician to select patients in need of therapy using the methods and compounds of the present invention.

In one embodiment the subject is a patient diagnosed with a condition, disease or disorder, or symptoms associated with a condition, disease or disorder, which condition, disease or disorder is associated with decreased expression of frataxin. In one embodiment the subject is a patient diagnosed with a condition, disease or disorder, or symptoms associated with a condition, disease or disorder, which condition, disease or disorder is associated with decreased expression of frataxin when compared to a healthy subject or patient. In one embodiment the subject is a patient diagnosed with a condition, disease or disorder, or symptoms associated with a condition, disease or disorder, which condition, disease or disorder is associated with a decreased level of frataxin. In one embodiment the subject is a patient diagnosed with a condition, disease or disorder, or symptoms associated with a condition, disease or disorder, which condition, disease or disorder is associated with a decreased level of frataxin when compared to a healthy subject or patient. In one embodiment the subject is a patient diagnosed with a condition, disease or disorder, which condition, disease or disorder is associated with decreased frataxin expression, but which patient or subject is asymptomatic. In one embodiment the subject is a patient diagnosed with a condition, disease or disorder, which condition, disease or disorder is associated with a decreased level of frataxin, but which patient or subject is asymptomatic. In one embodiment the subject is a patient diagnosed with Friedreich's ataxia. In one embodiment the subject is a patient diagnosed with Friedreich's ataxia but is asymptomatic. In one embodiment the subject is a patient diagnosed with Friedreich's ataxia with retained reflexes. In one embodiment the subject is a patient diagnosed with Friedreich's ataxia with retained reflexes but is asymptomatic. In one embodiment the subject is a patient diagnosed with late onset Friedreich's ataxia. In one embodiment the subject is a patient diagnosed with late onset Friedreich's ataxia but is asymptomatic. In one embodiment the subject is asymptomatic but has one or more genetic risk factors associated with a condition, disease or disorder, which condition, disease or disorder is associated with decreased frataxin expression. In one embodiment the subject is asymptomatic but has one or more genetic risk factors associated with a condition, disease or disorder, which condition, disease or disorder is associated with a decreased level of frataxin. In one embodiment the subject is asymptomatic but has one or more genetic risk factors associated with Friedreich's ataxia. In one embodiment the subject is asymptomatic but is judged to be at high risk of developing a condition, disease or disorder associated with decreased frataxin expression. In one embodiment the subject is asymptomatic but is judged to be at high risk of developing a condition, disease or disorder associated with a decreased level of frataxin. In one embodiment the subject is asymptomatic but is judged to be at high risk of developing Friedreich's ataxia. In one embodiment the subject is asymptomatic but is judged to be a carrier of Friedreich's ataxia.

In one embodiment the patient or subject has been identified as having greater than about 75 unstable homozygous GAA trinucleotide repeat expansions in the intron- 1 on both of the alleles in the gene which encodes frataxin. In one embodiment the patient or subject has been identified as having greater than about 75 unstable homozygous GAA trinucleotide repeat expansions in the intron-1 on one of the alleles in the gene which encodes frataxin. In one embodiment the patient or subject has been identified as having between about 75 and about 300 GAA triplet repeat expansions in the intron-1 on both of the alleles in the gene which encodes frataxin. In one embodiment the patient or subject has been identified as having between about 75 and about 300 GAA triplet repeat expansions in the intron-1 on one of the alleles in the gene which encodes frataxin. In one embodiment the patient or subject has been identified as having greater than about 300 GAA triplet repeat expansions in the intron-1 on both of the alleles in the gene which encodes frataxin. In one embodiment the patient or subject has been identified as having greater than about 300 GAA triplet repeat expansions in the intron-1 on one of the alleles in the gene which encodes frataxin. In one embodiment the subject or patient has been identified as being compound heterozygous for a GAA repeat expansion in the intron-1 of the gene which encodes frataxin and a point mutation or deletion.

In one embodiment the "patient" or "subject" with the condition, disease or disorder, or the symptoms associated with the condition, disease or disorder, has either decreased frataxin expression or a decreased level of frataxin such that the residual cellular level of frataxin is from about 2% to about 30%; more typically from about 5% to about 25%; more typically from about 10% to about 20% of the residual cellular level of frataxin of a healthy counterpart. As used herein the term "Friedreich's ataxia" shall be taken to mean an autosomal recessive congenital ataxia caused by a mutation(s) in the gene which encodes frataxin. In one embodiment, "Friedreich's ataxia" refers to a subject or patient who has been identified as having greater than about 300 unstable homozygous GAA trinucleotide repeat expansions in the intron-1 on both of the alleles in the gene which encodes frataxin. In one embodiment, "Friedreich's ataxia" refers to a subject or patient who has been identified as having late stage on-set of Friedreich's ataxia. In one embodiment, late stage on-set Friedreich's ataxia shall be taken to refer to a subject or patient who has greater than from about 75 to about 300 unstable homozygous GAA trinucleotide repeat expansion in the intron-1 on both of the alleles in the gene which encodes frataxin. In one embodiment "Friedreich's ataxia" refers to a subject or patient who has been identified as a heterozygous carrier of Friedreich's ataxia. In one embodiment, a carrier of Friedreich's ataxia refers to a subject or patient who has greater than about 75 unstable homozygous GAA trinucleotide repeat expansion in the intron-1 on one of the alleles in the gene which encodes frataxin. In one embodiment the subject or patient has been identified as being compound heterozygous for a GAA repeat expansion in the intron-1 of the gene which encodes frataxin and a point mutation or deletion.

In one embodiment the symptoms associated with the condition, disease or disorder are selected from the group consisting of muscle weakness in the arms and legs, loss of co-ordination, loss of deep tendon reflexes, loss of extensor plantar responses, loss of vibratory and proprioceptive sensation, vision impairment, involuntary and / or rapid eye movements, hearing impairment, slurred speech, curvature of the spine (scoliosis), high plantar arches (pes cavus deformity of the foot), carbohydrate intolerance, diabetes mellitus and heart disorders (including but not limited to atrial fibrillation, tachycardia (fast heart rate), hypertrophic cardiomyopathy, cardiomegaly, symmetrical hypertrophy, heart murmurs, and heart conduction defects).

As used herein, the term "mammal" includes, but is not limited to mammals such as a human, a non-human primate, canine, feline, bovine, ovine, porcine, murine, or other veterinary or laboratory mammals such as rabbits, rats, and mice. Those skilled in the art recognize that a therapy which reduces the severity of pathology in one species of mammal is predictive of the effect of the therapy on another species of mammal. As used herein, the term "modulate" shall be taken to encompass either a decrease or an increase in the expression, level or activity depending on the target molecule.

As used herein the term "regulating the expression of" shall be taken to mean either an intervention which results in a decrease in the expression of the target molecule, or an intervention which results in an increase in the expression of the target molecule. In one embodiment the regulation is an intervention which results in an "increase" or "up-regulation", which terms are used herein interchangeably, and which, as used herein, shall be taken to mean resulting in, either directly or indirectly, an increase in the level of the target molcule expressed.

As used herein, the term "increasing the expression of frataxin" shall be taken to mean that the cellular expression of frataxin precursor has, either directly or indirectly, been increased when compared to the cellular expression of frataxin precursor prior to treatment in accordance with the method of the invention. Without wishing to be bound by theory, it is thought that affecting an increase in the expression of frataxin precursor will result initially in an increase in the level of frataxin precursor which will in turn lead to an increase in the level of one or both of intermediate frataxin and mature frataxin as the frataxin precursor is further metabolized. Furthermore, without wishing to be bound by theory, in one embodiment it is thought that the expression of frataxin precursor is directly increased by modulation of the epigenetic state of the gene which encodes frataxin for example by either direct or indirect downstream modulation of P-TEFb and RNA polymerase II; by modulation of histone methylation; by modulation of transcriptional pause release; by relieving repression; or by modulating the methylation of CPG's in the DNA. Alternatively it is possible that the expression of frataxin precursor is directly increased by the modulation of the epigenetic state via relieving heterochromatinisation. In another embodiment, it is possible that the expression of frataxin precursor is modulated indirectly for example by inhibiting the expression of a negative regulator of frataxin, for example the negative anti-sense oligo repressor of expression FAST- 1 , or by inducing the expression of a positive regulator of frataxin. For example, without wishing to be bound by theory, epigenetic silencing in Friedreich's ataxia may be associated with depletion of CTCF (CCCTC-binding factor) and antisense transcription.

As used herein, the term "increasing the level of frataxin" shall be taken to mean that the level of frataxin, including frataxin precursor, intermediate frataxin and / or mature frataxin, in a cell, subject or patient has, either directly or indirectly, been increased when compared to the level of frataxin, including frataxin precursor, intermediate frataxin and / or mature frataxin, in a cell subject or patient prior to treatment in accordance with the method of the invention. Without wishing to be bound by theory, one means for directly increasing the level of frataxin, including frataxin precursor, intermediate frataxin and / or mature frataxin, in a cell, subject or patient is to increase the expression of frataxin. Alternatively, without wishing to be bound by theory, means for indirectly increasing the level of frataxin, including frataxin precursor, intermediate frataxin and / or mature frataxin, in a cell, subject or patient may include either increasing the stability of frataxin mRNA and / or decreasing the rate of degradation of frataxin, including frataxin precursor, intermediate frataxin, and / or mature frataxin.

In one embodiment the level of frataxin, including frataxin precursor, intermediate frataxin and / or mature frataxin, is increased by at least about 1.5 fold when compared to the level of frataxin, including frataxin precursor, intermediate and / or mature frataxin in the cell, subject or patient prior to treatment in accordance with a method of the invention. In one embodiment the level of frataxin, including frataxin precursor, intermediate frataxin and / or mature frataxin, is increased by at least about 2 fold when compared to the level of frataxin, including frataxin precursor, intermediate frataxin and / or mature frataxin, in the cell, subject or patient prior to treatment in accordance with a method of the invention. In one embodiment the level of frataxin, including frataxin precursor, intermediate frataxin and / or mature frataxin, is increased by at least about 5 fold when compared to the level of frataxin, including frataxin precursor, intermediate frataxin and / or mature frataxin, in the cell, subject or patient prior to treatment in accordance with a method of the invention. In one embodiment, the level of frataxin, including frataxin precursor, intermediate frataxin and / or mature frataxin, is measured as the steady state level after a period of treatment.

In one embodiment the level of frataxin, including frataxin precursor, intermediate frataxin and / or mature frataxin, in the cell, subject or patient treated in accordance with a method of the invention is increased to at least about 10% of the level of frataxin, including frataxin precursor, intermediate frataxin and / or mature frataxin, in a healthy cell or a healthy subject. In another embodiment the level of frataxin, including frataxin precursor, intermediate frataxin and / or mature frataxin, in the cell, subject or patient treated in accordance with a method of the invention is increased to at least about 20% of the level of frataxin, including frataxin precursor, intermediate frataxin and / or mature frataxin, in a healthy cell or a healthy subject. In another embodiment the level of frataxin, including frataxin precursor, intermediate frataxin and / or mature frataxin, in the cell, subject or patient treated in accordance with a method of the invention is increased to at least about 30% of the level of frataxin, including frataxin precursor, intermediate frataxin and / or mature frataxin, in a healthy cell or a healthy subject. In another embodiment the level of frataxin, including frataxin precursor, intermediate frataxin and / or mature frataxin, in the cell, subject or patient treated in accordance with a method of the invention is increased to at least about 40% of the level of frataxin, including frataxin precursor, intermediate frataxin and / or mature frataxin, in a healthy cell or a healthy subject. In another embodiment the level of frataxin, including frataxin precursor, intermediate frataxin and / or mature frataxin, in the cell, subject or patient treated in accordance with a method of the invention is increased to at least about 50% of the level of frataxin, including frataxin precursor, intermediate frataxin and / or mature frataxin, in a healthy cell or a healthy subject. In another embodiment the level of frataxin, including frataxin precursor, intermediate frataxin and / or mature frataxin, in the cell, subject or patient treated in accordance with a method of the invention is increased to at least about 75% of the level of frataxin, including frataxin precursor, intermediate frataxin and / or mature frataxin, in a healthy cell or a healthy subject. In another embodiment the level of frataxin, including frataxin precursor, intermediate frataxin and / or mature frataxin, in the cell, subject or patient treated in accordance with a method of the invention is increased to at least about 100% of the level of frataxin, including frataxin precursor, intermediate frataxin and / or mature frataxin, in a healthy cell or a healthy subject.

In one embodiment, the level of frataxin, including frataxin precursor, intermediate frataxin and / or mature frataxin, in the cell, subject or patient treated in accordance with a method of the invention is increased to about that of a heterozygous carrier of Friedreich's ataxia. In another embodiment, the level of frataxin, including frataxin precursor, intermediate frataxin and/or mature frataxin, in the cell, subject or patient treated in accordance with a method of the invention is increased to more than about that of a heterozygous carrier of Friedreich's ataxia. In another embodiment, the level of frataxin, including frataxin precursor, intermediate frataxin and/or mature frataxin, in the cell, subject or patient treated in accordance with a method of the invention is increased to be about the same as that of a healthy cell or subject. In one embodiment, the level of frataxin, including frataxin precursor, intermediate frataxin and/or mature frataxin, is measured as a steady state level achieved after a period of treatment.

In one embodiment, the relative or absolute increase in level of frataxin is determined by comparing the total amount of frataxin, including frataxin precursor, intermediate frataxin and mature frataxin, present in the cell, subject, or patient before and after treatment in accordance with a method of the invention. In another embodiment, the relative or absolute increase in level of frataxin is determined by comparing the amount of mature frataxin present in the cell, subject, or patient before and after treatment in accordance with a method of the invention. In another embodiment, the relative or absolute increase in level of frataxin is determined by comparing the amount of frataxin precursor present in the cell, subject, or patient before and after treatment in accordance with a method of the invention. In one embodiment, the relative or absolute increase in the level of frataxin is achieved by an increase in the expression of frataxin and is determined by comparing the amount of frataxin precursor present in the cell, subject or patient before and after treatment in accordance with a method of the invention. In another embodiment, the relative or absolute increase in the level of frataxin is achieved by an increase in the expression of frataxin and is determined by comparing the amount of mature frataxin present in the cell, subject or patient before and after treatment in accordance with a method of the invention.

Non-limiting examples of useful increases in the levels of frataxin are provided.

For example, without wishing to be bound by theory, it is believed from observing patients or subjects with greater than from about 75 GAA triplet repeats to about 300 GAA triplet repeats, that a level of frataxin, including frataxin precursor, intermediate frataxin and / or mature frataxin, of about 20% of the level of frataxin, including frataxin precursor, intermediate frataxin and / or mature frataxin, of that seen in a healthy cell or subject results in late onset of the condition, disease or disorder, or the symptoms associated with the condition, disease or disorder. As such, it is expected that where a method of the present invention results in an increase in the level of frataxin, including frataxin precursor, intermediate frataxin and / or mature frataxin, to greater than about 20% of the level in a healthy cell or subject, the increase in frataxin, including frataxin precursor, intermediate frataxin and / or mature frataxin, is expected to slow the advancement of the condition, disease or disorder or the symptoms associated with the condition, disease or disorder. By way of further example, and again not wishing to be bound by theory, it is believed from observing heterozygous carriers that a level of about 50% of the level of frataxin, including frataxin precursor, intermediate frataxin and / or mature frataxin, of that seen in a healthy cell or subject results in asymptomatic state. As such, it is expected that where a method of the present invention results in an increase in the level of frataxin, including frataxin precursor, intermediate frataxin and / or mature frataxin, to greater than about 50% of the level in a healthy cell or subject, the increase in frataxin, including frataxin precursor, intermediate frataxin and / or mature frataxin, is expected to significantly slow the advancement of the condition, disease or disorder or the symptoms associated with the conditions, disorder or disease, and potentially even prevent further advancement or lead to regression of the condition disorder or disease, or the symptoms associated with the condition, disease or disorder.

As used herein the term "healthy cell" is taken to mean a cell taken from a "healthy subject".

As used herein, the term "healthy subject" shall be taken to mean a subject who has not been diagnosed with a condition, disease or disorder associated with decreased level of frataxin or with decreased expression of frataxin. In one embodiment the healthy subject has been identified as not having Friedreich's ataxia. In another embodiment the healthy subject has been identified as not having late onset Friedreich's ataxia. In one embodiment, the healthy subject has been identified as having less than about 75 unstable homozygous GAA trinucleotide repeat expansion in the intron-1 on both of the alleles in the gene which encodes frataxin. In one embodiment, the healthy subject has been identified as having less than about 25 unstable homozygous GAA trinucleotide repeat expansion in the intron-1 on both of the alleles in the gene which encodes frataxin. In one embodiment, the healthy subject has been identified as not having pathological mutations or repeat expansions on either allele of the intron-1 of the gene which encodes frataxin.

As used herein the term "decreased level of frataxin" shall be taken to mean that the level of frataxin, including frataxin precursor, intermediate frataxin and mature frataxin, present in a cell, subject or patient is lower than the level of frataxin, including frataxin precursor, intermediate frataxin and / or mature frataxin, observed in a healthy cell or subject. In one embodiment the decreased level of frataxin refers to a decreased level of frataxin precursor. In one embodiment the decreased level of frataxin refers to a decreased level of mature frataxin.

As used herein the term "decreased expression of frataxin" shall be taken to mean that the expression of frataxin, including frataxin precursor, intermediate frataxin and / or mature frataxin, in a cell, subject or patient is lower than the expression of frataxin, including frataxin precursor, intermediate frataxin and mature frataxin, observed in a healthy cell or subject. In one embodiment the decreased expression of frataxin refers to a decreased expression of frataxin precursor.

In one embodiment the cells associated with the methods of the present invention are those where a decrease in frataxin expression or a decrease in the level of frataxin in one or more of these cell types is thought to contribute specifically, in whole or in part, to the in vivo pathology of the patient or subject with the condition, disease or disorder associated with decreased frataxin expression or a decreased level of frataxin, or the associated symptoms. Such cells include, but are not limited to, cerebellar neurons, neurons of the spinal tract, dorsal root ganglia and sensory neurons, aural and optic nerves, cardiomyocytes, and / or cells of the pancreas including islets. Without wishing to be bound by theory, cellular pathways believed to be modulated by changes in levels of frataxin include, but are not limited to, pathways that rely on functionality of an iron-sulphur protein such as the enzyme aconitase in the Kreb's/TCA cycle; succinate dehydrogenase subunit B (SDHB) in the electron transport chain; DNA polymerases (Pol α, δ and ε) and the major DNA mutagenesis enzyme Pol ζ important in DNA replication mechanisms and xanthine oxidase used in purine catabolism.

In one embodiment the cells associated with the methods of the invention are obtained from, or are present in, a subject diagnosed with a condition, disease or disorder associated with decreased expression of frataxin. In one embodiment the cell is obtained from, or is present in, a subject diagnosed with a condition, disease or disorder associated with a decreased level of frataxin. In one embodiment the cell is obtained from, or is present in, a subject diagnosed with Friedreich's ataxia. In one embodiment the cell is obtained from, or is present in, a human subject diagnosed with Friedreich's ataxia. In another embodiment the cell is obtained from, or is present in, a non-human species diagnosed with either a condition, disease or disorder associated with decreased expression of frataxin or condition, disease or disorder associated with a decreased level of frataxin or with Friedreich's ataxia, including genetic variants thereof. In one embodiment the cell is in vivo. In another embodiment the cell is ex vivo. In another embodiment the cell is in vitro.

As used herein the term "frataxin" shall be taken to mean one or more of the precursor polypeptide, intermediate frataxin and mature frataxin and the frataxin encoding nucleotide sequence and post translationally modified protein or polypeptide polymorphic variants, spliced variants, alleles, mutants and interspecies homologs that: (1) have an amino acid sequence that has greater than about 90% amino acid sequence identity, for example 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% or greater amino acid sequence identity, preferably over a region of at least about 25, 50, 100, 200, 300, 400 or more amino acids or over the full-length to an amino acid sequence encoded by frataxin nucleic acid or to an amino acid sequence encoded by a frataxin polypeptide; (2) bind to antibodies eg polyclonal antibodies, raised against an immunogen comprising an amino acid sequence encoded by a frataxin nucleic acid (eg frataxin polynucleotides described herein), and conservatively modified variants thereof; (3) specifically hybridise under stringent hybridization conditions to an anti-sense strand corresponding to a nucleic acid sequence that has greater than about 90% etc.

As used herein the term "gene which encodes frataxin" refers to a genomic region that encodes frataxin protein and / or controls the transcription of frataxin mRNA. Thus, the term shall be taken to encompass coding sequences as well as any non- coding elements, including but not limited to, promoters, enhancers, silencers, introns and 5' and 3' untranslated regions. A gene which encodes frataxin may include flanking sequences 5' and / or 3' to a known annotated frataxin open reading frame, including, but not limited to, 1 Kb, 2Kb, 3Kb, 4Kb, 5Kb, 6Kb, 7Kb, 8Kb, 9Kb, and / or 10Kb, or more flanking the 5' and / or 3' end of a known annotated frataxin open reading frame. In one embodiment, a gene which encodes frataxin may be a human gene which encodes frataxin. In an alternative embodiment, a gene which encodes frataxin may be a corresponding homolog for a gene which encodes frataxin in a different species, including, but not limited to, for example a mouse.

Methods are available in the art for determining the level of frataxin, including frataxin precursor, intermediate frataxin and / or mature frataxin for example routine methods such as Western Blot analysis or commercially available dip stick testing kits such as those available from Abeam^® or Mitosciences^®.

Methods are available in the art for the identification of conditions, diseases or disorders associated with a decreased expression of frataxin; conditions, diseases or disorders associated with a decreased level of frataxin; for the identification of GAA triplet repeat expansions on the intron-1 of the gene which encodes frataxin; and / or which identify a point mutation or deletion on the gene which encodes frataxin including methods for the diagnosis of Friedreich's ataxia, for example the Friedreich's ataxia repeat expansion test available from Athena Diagnostics^®.

As used herein, the term "BET-family bromodomain" refers to members of the bromodomain family of proteins which contain two N-terminal bromodomains. This family includes bromodomain members known as BRD2, BRD3, BRD4 and BRDT. These bromodomains are well known to share a common domain architecture featuring two amino-terminal bromodomains which exhibit high levels of sequence conservation, and a more divergent carboxy terminal recruitment domain.

As used herein, the term "BRD4" refers to BET-family bromodomain-containing protein 4, which is a member of the BET bromodomain family.

As used herein, the term "BET-family bromodomain inhibitor" shall be taken to mean a compound which shows inhibitory activity at one or more of the BET-family bromodomains, including by preventing the bromodomain from binding to acetyl- modified histone tails. In one embodiment the "BET-family bromodomain inhibitor" is a BRD4 inhibitor. In one embodiment the "BET-family bromodomain inhibitor" is a compound which inhibits bromodomain 1 of BRD4. In one embodiment the "BET-family bromodomain inhibitor" is a compound which inhibits bromodomain 2 of BRD4. In one embodiment the "BET-family bromodomain inhibitor" is a compound which inhibits both bromodomain 1 and bromodomain 2 of BRD4. Routine methods are available in the art to determine BET-family bromodomain activity such as the fluorescence binding assay disclosed herein and others.

As used herein, the term "IC50" refers to an amount, concentration or dosage of a particular test compound that achieves a 50% inhibition of a maximal response in an assay that measures such response. The value depends on the assay used.

As used herein, the term "other therapeutic agents" as used herein, refers to any therapeutic agent that has been used, is currently used or is known to be useful for treating a disease or a disorder encompassed by the present invention.

BET-family bromodomain inhibitors

The pharmaceutical methods and associated methods of treatment and pharmaceutical compositions of the present invention utilize BET-family bromodomain inhibitors, or a pharmaceutically acceptable salt thereof.

Bromodomain-containing proteins are known to be of substantial biological interest, as components of transcription factor complexes and determinants of epigenetic memory. The BET-fami!y (BRD2, BRD3, BRD4 and BRDT) shares a common domain architecture featuring two amino-terminal bromodomains that exhibit high levels of sequence conservation, and a more divergent carboxy-terminal recruitment domain (Filippakopou!os, P, et al., Nature 2010, 468, 1067-1073, incorporated by reference herein in its entirety for all purposes). BRD2 and BRD3 are reported to associate with histones along actively transcribed genes and may be involved in facilitating transcriptional elongation (Leroy et al, Mol. Cell. 2008, 30, 51 -60, incorporated by reference herein in its entirety for all purposes). It has also been reported that BRD4 or BRD3 may fuse with NUT (nuclear protein in testis) forming novel fusion oncogenes, BRD4-NUT or BRD3-NUT, in a highly malignant form of epithelial neoplasia (French et al. Cancer Res., 2003, 63, 304-307 and French et al. J. Clin. Oncol. 2004, 22, 4135-4139, both of which are incorporated by reference herein in their entirety for all purposes). Data suggests that BRD-NUT fusion proteins contribute to carcinogensesis (French et al. Oncogene 2008, 27, 2237-2242, incorporated by reference herein in its entirety for ail purposes). BRDT is believed to be uniquely expressed in the testes and ovary. All family members have been reported to have some function in controlling or executing aspects of the cell cycle, and have been shown to remain in complex with chromosomes during cell division— suggesting a role in the maintenance of epigenetic memory. In addition some viruses make use of these proteins to tether their genomes to the host cell chromatin, as part of the process of viral replication (You et al. Cell 2004 117, 349-60, incorporated by reference herein in its entirety for all purposes). BRD4 appears to be involved in the recruitment of the pTEF-P complex to inducible genes, resulting in phosphorylation of RNA polymerase and increased transcriptional output (Hargreaves et al. Cell 2009 138, 129-145, incorporated by reference herein in its entirety for all purposes).

Bromodomain-containing protein 4 (BRD4) is a member of the BET-family that in yeast and animals contains two tandem bromodomains (BDI and BDII) and an extraterminal (ET) domain. BRD4 is a double bromodomain-containing protein that binds preferentially to acetylated chromatin. In humans, four BET proteins (BRD2, BRD3, BRD4 and BRDT) exhibit similar gene arrangements, domain organizations, and some functional properties (Wu, S. et al. J. Biol. Chem. 2007, 282, 13141-13145, incorporated by reference herein in its entirety for all purposes).

Small molecule BET-family bromodomain inhibitors have been described with a wide variety of different chemical scaffolds and different chemotype approaches are known to be useful (see for example Gamier et al. Expert Opinion Ther Patents 2014, 24(2), 185-199; Hewings et al. J Med Chem 2012, 55, 9393-413; and Owen et al. Drug Disco Today: Technologies 2012, 10, 1016, all of which are incorporated by reference herein in their entirety for ail purposes). To date, small molecule BET-family bromodomain inhibitors have been mainly reported as being useful for treatment inflammatory diseases and / or various cancers, but some have also been reported to be useful for the treatment of cardiovascular diseases and to treat male fertility. BET- family bromodomain inhibitors can be identified using routine methods available in the art such as the fluorescence polarization (FP) binding assay disclosed herein. In one embodiment of the present invention, the BET-family bromodomain inhibitors useful in the present invention have an activity of about≤5μΜ in the fluorescence polarization (FP) binding assay at either bromodomain 1 , bromodomain 2 or both bromodomain 1 and 2. In another embodiment, the BET-family bromodomain inhibitors useful in the present invention have an activity of about≤2μΜ in the fluorescence polarization (FP) binding assay disclosed herein at either bromodomain 1 , bromodomain 2 or both bromodomain 1 and 2. In another embodiment, the BET-family bromodomain inhibitors useful in the present invention have an activity of about≤1 μΜ in the fluorescence polarization (FP) binding assay disclosed herein at either bromodomain 1 , bromodomain 2 or both bromodomain 1 and 2.

Examples of BET-family bromodomain inhibitors useful in the present invention include, but are not limited to compounds described below, including those which are in clinical development.

For example, small molecule BET-family bromodomain inhibitors with a diazepine chemical scaffold were described in International Patent application number PCT/EP2010/061518, published as WO201 1/054553 on 12^th May 201 1 which is incorporated by reference herein in its entirety for all purposes. This disclosure includes the compound I-BET762, and methods to prepare it:

I-BET762 is reported to be in clinical development for the treatment of various indications including acute & chronic lymphocytic leukemia; acute & chronic myelogenous leukemia; Hodgkin's & Non-Hodgkin's lymphoma; multiple myeloma; myeloproliferative neoplasms; NUT mideline carcinoma; hematological carcinoma & solid tumors; and atherosclerosis. Other disclosures relating to I-BET762 and analogues thereof include those described in International Patent application number PCT/EP2010/066697, published as WO2011/054845 on 12^th May 2011 ; International Patent application number PCT/EP2010/066696,published as WO201 1/054844 on 12^th May 201 1 ; International Patent application number PCT/EP2011/060179, published as WO2011/161031 on 29^th December 201 1 ; and Nicodeme E et al. Nature, 2010, 468, 1 119-23, all of which are incorporated by reference herein in their entirety for ail purposes.

Small molecule BET-family bromodomain inhibitors with a thienodiazapine chemical scaffold were described in International Patent application number PCT/US2011/036701 , published as WO2011/143669 on 17^th November 201 1 , which is incorporated by reference herein in its entirety for all purposes. This disclosure includes the compound JQ-1 , which exists in two steroisomeric forms JQ-1 (+) and JQ- 1 (-), and methods to prepare it:

Other disclosures relating to JQ-1 and analogues thereof include those described in International Patent application number PCT/US201 1/036647, published as WO2011/143651 on 17^th November 2011 ; International Patent application number PCT/US2011/036667, published as WO2011/143657 on 17^th November 2011 ; International Patent application number PCT/US201 1/036672, published as WO2011/143660 on 17^th November 201 1 ; and Filippakopoulos P et al. Nature, 2010, 468, 1067-73, all of which are incorporated by reference herein in their entirety for all purposes.

Other small molecule compounds with a thienodiazapine chemical scaffold were described in International Patent application number PCT/JP92/01 198, published as WO 1994/06801 on 31 ^st March 1994, which is incorporated by reference herein in its entirety for all purposes. This disclosure includes the racemic form of the compound now known as CPI-203, also known as CPI-267203, and methods to prepare it:

CPI-203 has since been reported to have BRD4 activity (Devaiah, B. N. et al. Proc Natl Acad Sci USA 2012, 109(18), 6927-32, which is incorporated by reference herein in its entirety for all purposes) and is reported to be in preclinical development for a variety of indications including cancer; hematological cancer; lymphoma; and Non-Hodgkin's lymphoma.

Yet further small molecule BET-family bromodomain inhibitors with thienodiazapine chemical scaffold were described in International Patent application number PCT/JP97/02817, published as WO1998/01 111 1 on 19^th March 1998, which is incorporated by reference herein in its entirety for all purposes. This disclosure includes the compound OTX-015, and methods to prepare it:

OTX015 is reported to be in clinical development for various indications including leukemia; cancer; Crohn's disease; ulcerative colitis; hematological cancer; acute lymphocytic leukemia; acute myelogenous leukemia; Non-Hodgkin's lymphoma; multiple myeloma; and breast, lung, non-small cell, pancreas, prostate tumors. Further related compounds with this chemical scaffold are described in International Patent application PCT/JP2006/310709, published as WO2006/129623 on 7^th December 2006; and International Patent application PCT/JP2008/-73864, published as WO 2009/084693 on 9^th July 2009, both of which are incorporated by reference herein in their entirety for all purposes.

Small molecule BET-family bromodomain inhibitors with a benzodiazepine chemical scaffold have been disclosed in International Patent application number PCT/EP2010/066696, published as WO2011/054844 on 12^th May 2011 , which is incorporated by reference herein in its entirety for all purposes. This disclosure includes the compound GW-841819X, and methods to prepare it:

GW-841819X is reported to be in preclinical development for various indications including dyslipidemia; cancer; and hematological neoplasm.

Small molecule BET-family bromodomain inhibitors with a thienoazepine chemical scaffold have been described in International Patent application number PCT/US2011/063046, published as WO2012/075383 on 7^th June 2012, which is incorporated by reference herein in its entirety for all purposes. This disclosure includes the compound CP-0610, and methods to prepare it:

CPI-0610 is reported to be in clinical development for various indications including Hodgkin's & Non-Hodgkin's lymphoma; multiple myeloma; acute lymphocytic leukemia; acute and chronic myelogenous leukemia; myelodysplasia syndrome; and myeloproliferative neoplasms.

Small molecule BET-family bromodomain inhibitors with an oxazolazepine chemical scaffold have been described in International Patent application number PCT/US2011/063046, published as WO2012/075383 on 7^th June 2012, which is incorporated by reference herein in its entirety for all purposes. This disclosure includes the compound CPI-232, and methods to prepare it:

Small molecule BET-family bromodomain inhibitors with a benzoazepine chemical scaffold have been described in International Patent application number PCT/EP2013/069902, published as WO2014/048945 on 3^rd April 2014, which is incorporated by reference herein in its entirety for all purposes. This disclosure includes the compound BET-BAY-002, and methods to prepare it:

BET-BAY-002 is reported to be in preclinical development for various indications including cancer.

Small molecule BET-family bromodomain inhibitors with a quinolone chemical scaffold have been described in International Patent application number PCT/EP2010/066695, published as WO201 1/054843 on 12^th May 2011 , which is incorporated by reference herein in its entirety for all purposes. This disclosure includes the compound l-BET-151 , also known as GSK1210151 , and methods to prepare it:

l-BET-151 is reported to be in preclinical development for various indications including leukemia; cancer; mixed lineage leukemia; hematological cancer; bone resorprtion; and inflammatory disease.

Small molecule BET-family bromodomain inhibitors with a quinazoline chemical scaffold have been described in International Patent application number PCT/CA2007/000146, published as WO2008/092231 on 7^th August 2008, which is incorporated by reference herein in its entirety for all purposes. This disclosure includes the compound RVX-208, and methods to prepare it:

RVX-208 is reported to be in clinical development for various indications including atherosclerosis; diabetes; acute coronary syndrome; myocardial Infarction; Alzheimer's disease; stroke; mild cognitive impairment; low HDL cholesterol; and immunological diseases.

Small molecule BET-family bromodomain inhibitors with a benzopiperazine chemical scaffold have been described in International Patent application number PCT/EP2010/066695, published as WO2011/054843 on 12^th May 2011 , which is incorporated by reference herein in its entirety for all purposes. This disclosure includes the compound l-BET-726, also known as GSK1324726A, and methods to prepare it:

l-BET-726 is reported to be in preclinical development for various indications including cancer.

Small molecule BET-family bromodomain inhibitors with a thiophenpyran chemical scaffold have been described in International Patent application number PCT/US2009/031864, published as WO 2009/094560 on 30^th July 2009, which is incorporated by reference herein in its entirety for all purposes. This disclosure includes the compound SRX-2523, and methods to prepare it:

SRX-2523 is reported to be in preclinical development for various indications including cancer and hematological neoplasm.

BET-family bromodomain inhibitors of the following formula:

or pharmaceutically acceptable salts and solvates thereof, are disclosed in International Patent application PCT/IB2012/054211 , published as WO 2013/027168 on 28^th February 2013, which is incorporated by reference herein in its entirety for all purposes. BET-family bromodomain inhibitors disclosed therein include N-(2-hydroxy-3- methylquinolin-6-yl)piperidine-1-sulfonamide

or a pharmaceutically acceptable salt or solvate thereof, which is specifically exemplified as example 46 in the above mentioned publication.

BET-family bromodomain inhibitors of the following formula (I):

or pharmaceutically acceptable salts thereof, are disclosed in International Patent application PCT/CN2011/002224, published as WO 2013/097052 on 4^th July 2013; and International Patent application PCT/CN2012/086357, published as WO 2013/097601 on 4 July 2013, both of which are incorporated by reference herein in their entirety for all purposes. BET-family bromodomain inhibitors disclosed therein include N-[4-(2,4- difluorophenoxy)-3-(6-methyl-7-oxo-6,7-dihydro-1 H-pyrrolo[2,3-c]pyridin- 4- yl)phenyl]ethanesulfonamide:

or a pharmaceutically acceptable salt thereof, which is specifically exemplified in the above mentioned publications. Further related publications include International Patent application PCT/US2014/044513, published as WO 2014/210425 on 31^st December 2014, which is incorporated by reference herein in its entirety for all purposes.

Other BET-family bromodomain inhibitors which are useful for the methods disclosed herein include, but are not limited to, the following BET-family bromodomain inhibitors which have been disclosed in the public domain as being in preclinical or clinical development BMS-986158 (Bristol Myers Squibb); RVX-297 (Resverlogix); RVS-2135 (Resverlogix); ZEN-3365 (Zenith); ZEN-31 18 (Zenith); KM-601 (Kainos); SF- 2535 (Signal Rx); AU-004 (Aurigene, Orion); ABBV-075 (Abbvie); TEN-010 (Tensha Therapeutics); and BAY1238097 (Bayer).

Yet further BET-family bromodomain inhibitors which are useful for the methods described herein include, but are not limited to, compounds disclosed in the following international patent applications: triazolodiazepine derivatives disclosed in International Patent application PCT/US2012/036569, published as WO 2012/151512 on 8^th November 2012; tetrahydroquinoline derivatives disclosed in International Patent application PCT/EP2010/066701 , published as WO 2011/054848 on 12^th May 2011 ; imidazoquinoline derivatives disclosed in International patent application PCT/EP2010/066699, published as WO 201 1/054846 on 12^th May 201 1 ; quinazolinone derivatives disclosed in International Patent application PCT/US2009/048457, published as WO 2009/158404 on 30^th December 2009; quinazolinone derivatives disclosed in International Patent application PCT/US2010/031870, published as WO 2010/123975 on 28 October 2010; triazolopyridazine-6-amine derivatives disclosed in International Patent application PCT/US2012/042825, published as WO 2012/174487 on 20^th December 2012; N-methylpyrrolopyridinones and N-methylpyrrolopyridazinone derivatives disclosed in International Patent application PCT/CN2012/086357, published as WO 2013/097601 on 4^th July 2013; and bis aryl derivatives described in International Patent application PCT/US2010/026308, published as WO 2012/1 16170 on 30^th August 2012, all of which are incorporated by reference herein in their entirety for all purposes.

Yet further BET -family bromodomain inhibitors suitable for use with the methods of the present invention include those described in US provisional patent application serial number 62/181281 , which was filed on 18^th June 2015 which is incorporated by reference herein in its entirety for all purposes. This disclosure includes compounds of Formula I:

Formula I

or a pharmaceutically acceptable salt thereof, wherein

is selected from the group consisting of:

-C3-C₇cycloalkyl optionally substituted with one, two, three or four E;

4 to 7 membered heterocyclyl optionally substituted with one, two, three or four E, which said 4 to 7 membered heterocyclyl comprises one or two heteroatoms independently selected for each occurrence from the group of N, O and S; and

R^1A is selected from the group consisting of

(i) -d-Cealkyl optionally substituted with one, two, three, four, five or six E; (ii) -C3-C₇cycloalkyl optionally substituted with one, two, three, four or five E;

(iii) phenyl optionally substituted with one, two, three, four or five E;

(iv) 4 to 7 membered heterocyclyl optionally substituted with one, two, three, four or five E, which said 4 to 7 membered heterocyclyl comprises one or two heteroatoms

independently selected for each occurrence from the group consisting of N, O and S; and

(v) 5 to 6 membered heteroaryl optionally substituted with one, two, three, four or five E, which said 5 to 6 membered heteroaryl comprises one, two or three heteroatoms independently selected for each occurrence from the group consisting of N, O and S;

R^{1 B} is selected from the group consisting of

(i) -H; and

(ii) -d-Cealkyl optionally substituted with one, two, three, four, five or six E;

R^{1 C} is selected from the group consisting of

(i) -H;

(ii) -CH₃ optionally substituted with one, two, or three J;

(iii) -CH2CH3 optionally substituted with one, two, three, four or five J;

(iv) -CH2CH2CH3 optionally substituted with one, two, three, four, five, six or seven J; and

(v) -CH(CH3)2 optionally substituted with one, two, three, four, five, six or seven J;

R^2A is selected from the group consisting of

(i) -H;

(ii) -CH3 optionally substituted with one, two or three J;

(iii) -CH2CH3 optionally substituted with one, two, three, four, or five J; and

(iv) cyclopropyl optionally substituted with one, two, three, four or five J;

R^2B is selected from the group consisting of (i) -d-Cealkyl optionally substituted with one, two, three or four G;

(ii) -OCi-C₆alkyl optionally substituted with one, two, three or four G;

(iii) -IMH2;

(iv) -NH(Ci-C₆alkyl), which Ci-Cealkyl is optionally substituted with one, two, three or four G;

(v) -N(CrC₆alkyl)₂, which d-Cealkyl is, independently for each

occurrence, optionally substituted with one, two, three or four G;

(vi) C3-C₅cycloalkyl optionally substituted with one, two, three or four G; and

(vii) 4 to 7 membered heterocyclyl optionally substituted with one, two, three or four G, which said 4 to 7 membered heterocyclyl comprises one or two heteroatoms independently selected for each occurrence from the group consisting of N, O and S;

W is selected from the group consisting of:

-?-o 54 A

(ii)

"Y _D4A

(iii)

->-0^_R4C

(v) 5 ; and

(vi) 4 to 7 membered heterocyclyl optionally substituted with one, two, three or four G, which said 4 to 7 membered heterocyclyl comprises one, two, three or four heteroatoms independently selected for each occurrence from the group consisting of N, O and S;

Y is selected from the group consisting of:

(i) -CH₂- optionally substituted with one or two J;

(ii) -(CH₂)₂- optionally substituted with one, two, three or four J; (iii) -(CH₂)3- optionally substituted with one, two, three, four, five or six J; and

(iv) -(CH2)4- optionally substituted with one, two, three, four, five, six, seven or eight J; R³ is selected from the group consisting of:

(i) -H;

(ii) -CH3 optionally substituted with one, two, or three J;

(iii) -CH2CH3 optionally substituted with one, two, three, four or five J;

R^4A is selected from the group consisting of

(i) -H;

(ii) -Ci-C6alkyl optionally substituted with one, two, three or four G;

(iv) -C(0)Ci-C6alkyl optionally substituted with one, two, three or four

G;

(v) -C(0)OCrC6alkyl optionally substituted with one, two, three or four G;

(vi) -C(0)NH₂;

(vii) -C(0)NH(Ci-C6alkyl) optionally substituted with one, two, three or four G;

(viii) -C(0)N(CrC6alkyl)₂ optionally substituted with one, two, three or four G;

(ix) -C(0)NHSC>2Ci-C3alkyl optionally substituted with one, two, three or four G;

(x) -NH(CrC3alkyl) optionally substituted with one, two, three or four

G;

(xi) -N(CrC3alkyl)₂ optionally substituted with one, two, three or four

G;

(xii) -NHC(0)Ci-C3alkyl optionally substituted with one, two, three or four G; (xiii) -N(Ci-C₃alkyl)C(0)CrC₃alkyl optionally substituted with one, two, three or four G;

(xiv) -NHSC>₂Ci-C₃alkyl optionally substituted with one, two, three or four G;

(xv) -N(Ci-C₃alkyl)SC>₂Ci-C₃alkyl optionally substituted with one, two, three or four G;

(xvii) -S0₂NH(Ci-C₃alkyl) optionally substituted with one, two, three or four G;

(xviii) -S0₂N(C C₃alkyl)₂ optionally substituted with one, two, three or four G;

(xix) -C₃-C₇cycloalkyl optionally substituted with one, two, three or four

G;

(xx) phenyl optionally substituted with one, two, three or four G;

(xxi) 4 to 7 membered heterocyclyl optionally substituted with one, two, three or four G, which said 4 to 7 membered heterocyclyl comprises one, two, three or four heteroatoms independently selected for each occurrence from the group consisting of N, O and S; and

(xxii) 5 to 6 membered heteroaryl optionally substituted with one, two, three or four G, which said 5 to 6 membered heteroaryl ring comprises one, two, three or four heteroatoms independently selected for each occurrence from the group consisting of N, O and

S;

elected from the group consisting of

(i) -H;

(ii) -d-Cealkyl optionally substituted with one, two, three or four G;

(iii) -C(0)Ci-C₆alkyl optionally substituted with one, two, three or four

G;

(iv) -C(0)OCrC₆alkyl optionally substituted with one, two, three or four

G;

(v) -C(0)NH₂;

(vi) -C(0)NH(Ci-C₆alkyl) optionally substituted with one, two, three or four G; (vii) -C(0)N(CrC₆alkyl)₂ optionally substituted with one, two, three or four G;

(viii) -C(0)NHSC>2Ci-C₃alkyl optionally substituted with one, two, three or four G;

(ix) -C3-C₇cycloalkyl optionally substituted with one, two, three or four

G;

(x) phenyl optionally substituted with one, two, three or four G;

(xi) 4 to 7 membered heterocyclyl optionally substituted with one, two, three or four G, which said 4 to 7 membered heterocyclyl comprises one, two, three or four heteroatoms independently selected for each occurrence from the group consisting of N, O and S; and

(xii) 5 to 6 membered heteroaryl optionally substituted with one, two, three or four G, which said 5 to 6 membered heteroaryl ring comprises one, two, three or four heteroatoms independently selected for each occurrence from the group consisting of N, O and

S;

is selected from the group consisting of

(i) -H;

(ii) -Ci-C₆alkyl optionally substituted with one, two, three or four G;

(iii) -C3-C₇cycloalkyl optionally substituted with one, two, three or four

G;

(iv) phenyl optionally substituted with one, two, three or four G;

(v) 4 to 7 membered heterocyclyl optionally substituted with one, two, three or four G, which said 4 to 7 membered heterocyclyl comprises one, two, three or four heteroatoms independently selected for each occurrence from the group consisting of N, O and S; and

(vi) 5 to 6 membered heteroaryl optionally substituted with one, two, three or four G, which said 5 to 6 membered heteroaryl ring comprises one, two, three or four heteroatoms independently selected for each occurrence from the group consisting of N, O and

S;

is independently selected for each occurrence from the group consisting of - F, -CI, -OH, -CN, -CH₃, -CH₂CH₃, -CH₂F, -CHF₂, -CF₃, -CF₂CF₃, -CH₂OH, - OCH₃, -OCH2F, -OCHF2, -OCF3, -SCH3, -SCH₂F, -SCHF2, -SCF3 -NH₂, - NH(CH₃), and -N(CH₃)₂;

E is independently selected for each occurrence from the group consisting of:

(i) -OH;

(ii) -CN;

(iii) -CO₂H;

(iv) -C(0)H;

(v) halo;

(vi) -Ci-C₃alkyl optionally substituted with one, two, three or four J;

(vii) -C C₃alkylCC>₂H which -d-Csalkyl is optionally substituted with one, two, three or four J;

(viii) -C₃-C₇cycloalkyl optionally substituted with one, two, three, four, five or six J;

(ix) -Ci-C₃alkylC₃-C₆cycloalkyl optionally substituted with one, two, three, four, five or six J;

(x) -OC C₃alkyl, optionally substituted with one, two, three or four J;

(xi) -OC₃-C₇cycloalkyl optionally substituted with one, two, three, four, five or six J;

(xii) -OCi-C₃alkylC₃-C₇cycloalkyl optionally substituted with one, two, three, four, five or six J;

(xiii) -SC C₃alkyl, optionally substituted with one, two, three or four J;

(xiv) -SC₃-C₇cycloalkyl optionally substituted with one, two, three, four, five or six J;

(xv) -SCi-C₃alkylC₃-C₇cycloalkyl optionally substituted with one, two, three, four, five or six J;

(xvi) -C(0)Ci-C₃alkyl, optionally substituted with one, two, three or four J;

(xvii) -C(0)OCrC₃alkyl, optionally substituted with one, two, three or four J;

(xviii) -NH₂;

(xix) -NH(CrC₃alkyl) optionally substituted with one, two, three or four J;

(xx) -N(CrC₃alkyl)₂ which -d-Csalkyl is, independently for each occurrence, optionally substituted with one, two, three or four J;

(xxi) -C(0)NH₂;

(xxii) -C(0)NHCrC₃alkyl, optionally substituted with one, two, three or four J;

(xxiii) -C(0)N(Ci-C₃alkyl)₂, which -d-Csalkyl is, independently for each

occurrence, optionally substituted with one, two, three or four J; (xxiv) -NHC(0)CrC₃alkyl, optionally substituted with one, two, three or four J;

(xxv) -SC>₂(Ci-C₃alkyl), optionally substituted with one, two, three or four J;

(xxvi) -SC>₂NH(Ci-C₃alkyl), optionally substituted with one, two, three or four J;

(xxvii) -NHSC>₂(Ci-C₃alkyl), optionally substituted with one, two, three or four J; and

(xxviii) phenyl optionally substituted with one, two, three, or four J;

G is independently selected for each occurrence from the group consisting of

(i) -OH;

(ii) -CN;

(iii) -CO₂H;

(iv) -C(0)H;

(v) halo;

(vi) -Ci-C₃alkyl, optionally substituted with one, two, three or four J;

(vii) -C C₃alkylCC>₂H, which -d-Csalkyl is optionally substituted with one, two, three or four J;

(viii) -Ci-C₃alkylC₃-C₆cycloalkyl optionally substituted with one, two, three, four, five or six J;

(ix) -OCi-C₃alkyl, optionally substituted with one, two, three or four J;

(x) -OCi-C₃alkylC₃-C₆cycloalkyl optionally substituted with one, two, three, four, five or six J;

(xi) -SC C₃alkyl, optionally substituted with one, two, three or four J;

(xii) -SCi-C₃alkylC₃-C₆cycloalkyl optionally substituted with one, two, three, four, five or six J;

(xiii) -C(0)Ci-C₃alkyl, optionally substituted with one, two, three or four J;

(xiv) -C(0)OCrC₃alkyl, optionally substituted with one, two, three or four J;

(xv) -IMH2;

(xvi) -NH(Ci-C₃alkyl), optionally substituted with one, two, three or four J;

(xvii) -N(Ci-C3alkyl)2, which -d-Csalkyl is, independently for each occurrence, optionally substituted with one, two, three or four J;

(xviii) -C(0)NH₂;

(xix) -C(0)NHCrC₃alkyl, optionally substituted with one, two, three or four J;

(xx) -C(0)N(CrC₃alkyl)₂, which -d-Csalkyl is, independently for each

occurrence, optionally substituted with one, two, three or four J;

(xxi) -NHC(0)Ci-C₃alkyl, optionally substituted with one, two, three or four J; (xxii) -S02(CrC3alkyl), optionally substituted with one, two, three or four J; (xxiii) -S0₂NH(Ci-C₃alkyl), optionally substituted with one, two, three or four J; and

(xxiv) -NHSC>₂(Ci-C₃alkyl) optionally substituted with one, two, three or four J; and

J is independently selected for each occurrence from the group consisting of -H, -F, -CI, -OH, -CN, -CH₃, -CH2CH3, -CH2F, -CHF2, -CF₃, -CF₂CF₃, -CH₂OH, - OCH3, -OCH2F, -OCHF2, -OCF3, -SCH3, -SCH₂F, -SCHF2, -SCF3 -NH₂, - NH(CH₃), and -N(CH₃)₂.

A preferred Embodiment of the compounds of Formula I disclosed in US provisional patent application serial number 62/181281 is also disclosed where:

R^1A is selected from the group consisting of -CH₂OCH₃; phenyl; methoxyphenyl; and pyridyl;

R^{1 B} is selected from the group consisting of methyl, ethyl, n-propyl, i-propyl, and

R^{1 C} is -H;

R^2A is selected from the group consisting of -CH3;

R^2B is selected from the group consisting of methyl, ethyl, n-propyl, i-propyl, and

-NH(CH₃);

Y is selected from the group consisting of -CH₂- and -CH₂CH₂-;

R³ is -H;

R^4A is selected from the group consisting of -CH₃; -C0₂H; and -C(0)NHCH₃; and

R¹⁰ is -H, or a pharmaceutically acceptable salt thereof. Specific examples of BET-family bromodomain inhibitors, including their methods of preparation, described in US provisional patent application serial number 62/181281 , which was filed on 18^th June 2015, include:

N-{6-[acetyl(methyl)amino]-4-[(1 S)-1-(2-methoxyphenyl)ethyl]-3-oxo-3,4- dihydropyrido[2,3-b]pyrazin-2-yl}-beta-alanine;

N-{6-[acetyl(ethyl)amino]-4-[(1 S)-1-(2-methoxyphenyl)ethyl]-3-oxo-3,4- dihydropyrido[2,3-b]pyrazin-2-yl}-beta-alanine;

N-{6-[acetyl(methyl)amino]-3-oxo-4-[(1 S)-1-phenylpropyl]-3,4-dihydropyrido[2,3- b]pyrazin-2-yl}-beta-alanine;

N-{6-[(hydroxyacetyl)(methyl)amino]-3-oxo-4-[(1 S)-1-phenylpropyl]-3,4- dihydropyrido[2,3-b]pyrazin-2-yl}-beta-alanine;

N-{4-[(1 S)-1-(2-methoxyphenyl)ethyl]-2-(methylamino)-3-oxo-3,4-dihydropyrido[2,3- b]pyrazin-6-yl}-N-methylacetamide;

N-{6-[methyl(2-methylpropanoyl)amino]-3-oxo-4-[(1 S)-1-phenylpropyl]-3,4- dihydropyrido[2,3-b]pyrazin-2-yl}-beta-alanine;

N-{6-[butanoyl(methyl)amino]-3-oxo-4-[(1 S)-1-phenylpropyl]-3,4-dihydropyrido[2,3- b]pyrazin-2-yl}-beta-alanine;

N-{6-[(cyclobutylcarbonyl)(methyl)amino]-3-oxo-4-[(1 S)-1-phenylpropyl]-3,4- dihydropyrido[2,3-b]pyrazin-2-yl}-beta-alanine;

N-{6-[methyl(methylcarbamoyl)amino]-3-oxo-4-[(1 S)-1-phenylpropyl]-3,4- dihydropyrido[2,3-b]pyrazin-2-yl}-beta-alanine;

N-{6-[methyl(propanoyl)amino]-3-oxo-4-[(1 S)-1-phenylpropyl]-3,4-dihydropyrido[2,3- b]pyrazin-2-yl}-beta-alanine;

N-{6-[methyl(propanoyl)amino]-3-oxo-4-[(1 S)-1-(pyridin-2-yl)propyl]-3,4- dihydropyrido[2,3-b]pyrazin-2-yl}-beta-alanine;

N-{6-[methyl(propanoyl)amino]-3-oxo-4-(1-phenylcyclobutyl)-3,4-dihydropyrido[2,3- b]pyrazin-2-yl}-beta-alanine;

N-{4-(2,5-diethylcyclopentyl)-6-[methyl(propanoyl)amino]-3-oxo-3,4-dihydropyrido[2,3- b]pyrazin-2-yl}-beta-alanine;

N-{6-[acetyl(methyl)amino]-4-[(1 f?)-2-methoxy-1 -phenylethyl]-3-oxo-3,4- dihydropyrido[2,3-b]pyrazin-2-yl}-beta-alanine;

N-{6-[methyl(propanoyl)amino]-3-oxo-4-[(3S,4S)-4-phenyltetrahydrofuran-3-yl]-3,4- dihydropyrido[2,3-b]pyrazin-2-yl}-beta-alanine;

N-{6-[acetyl(methyl)amino]-4-[(2f?)-1-methoxybutan-2-yl]-3-oxo-3,4-dihydropyrido[2,3- b]pyrazin-2-yl}-beta-alanine; N-{4-[(2f?)-1-methoxypentan-2-yl]-6-[methyl(propanoyl)amino]-3-oxo-3,4- dihydropyrido[2,3-b]pyrazin-2-yl}-beta-alanine;

N-(4-[(2 )-1-methoxybutan-2-yl]-2-{[2-(methylamino)-2-oxoethyl]arTiino}-3-oxo-3,4- dihydropyrido[2,3-b]pyrazin-6-yl)-N-methylpropanamide;

N-[4-(1 ,3-dimethoxypropan-2-yl)-2-{[2-(methylamino)-2-oxoethyl]arriino}-3-oxo-3,4- dihydropyrido[2,3-b]pyrazin-6-yl]-N-methylpropanamide;

N-[4-(1 ,3-dimethoxypropan-2-yl)-2-(methylamino)-3-oxo-3,4-dihydropyrido[2,3- b]pyrazin-6-yl]-N-methylpropanamide;

N-{2-(acetylamino)-3-oxo-4-[(1 S)-1-phenylpropyl]-3,4-dihydropyrido[2,3-b]pyrazin-6-yl}- N-methylacetamide;

N-{6-[(dimethylcarbamoyl)(methyl)amino]-3-oxo-4-[(1 S)-1-phenylpropyl]-3,4- dihydropyrido[2,3-b]pyrazin-2-yl}-beta-alanine;

N-{6-[methyl(propanoyl)amino]-4-[(1 S)-2-methyl-1-(pyridin-2-yl)propyl]-3-oxo-3,4- dihydropyrido[2,3-b]pyrazin-2-yl}-beta-alanine;

N-{4-(1-cyclopentylcyclopropyl)-6-[methyl(propanoyl)amino]-3-oxo-3,4- dihydropyrido[2,3-b]pyrazin-2-yl}-beta-alanine;

N-(6-{[(3,3-dimethylcyclobutyl)carbonyl](methyl)amino}-3-oxo-4-[(1 S)-1-phenylpropyl]- 3,4-dihydropyrido[2,3-b]pyrazin-2-yl)-beta-alanine;

N-(6-{[(3,3-difluorocyclobutyl)carbonyl](methyl)amino}-3-oxo-4-[(1 S)-1-phenylpropyl]- 3,4-dihydropyrido[2,3-b]pyrazin-2-yl)-beta-alanine;

N-{6-[methyl(oxetan-3-ylcarbonyl)amino]-3-oxo-4-[(1 S)-1-phenylpropyl]-3,4- dihydropyrido[2,3-b]pyrazin-2-yl}-beta-alanine;

N-methyl-N-(2-{[3-(methylamino)-3-oxopropyl]amino}-3-oxo-4-[(1 S)-1-phenylpropyl]-3,4- dihydropyrido[2,3-b]pyrazin-6-yl)oxetane-2-carboxamide;

N-methyl-N-(2-{[2-(methylamino)-2-oxoethyl]amino}-3-oxo-4-[(1 S)-1-phenylpropyl]-3,4- dihydropyrido[2,3-b]pyrazin-6-yl)propanamide;

N-methyl-N-(2-{[2-(methylamino)-2-oxoethyl]amino}-4-[(1 S)-2-methyl-1-(pyridin-2- yl)propyl]-3-oxo-3,4-dihydropyrido[2,3-b]pyrazin-6-yl)propanamide;

N³-{4-[(1f?)-2-methoxy-1-phenylethyl]-6-[methyl(propanoyl)amino]-3-oxo-3,4- dihydropyrido[2,3-b]pyrazin-2-yl}-N-methyl-beta-alaninamide;

N-methyl-N³-{6-[methyl(propanoyl)amino]-3-oxo-4-[(1 S)-1-phenylpropyl]-3,4- dihydropyrido[2,3-b]pyrazin-2-yl}-beta-alaninamide;

N-methyl-N³-{6-[methyl(propanoyl)amino]-3-oxo-4-[(1S)-1-(pyridin-2-yl)propyl]-3,4- dihydropyrido[2,3-b]pyrazin-2-yl}-beta-alaninamide; and N-methyl-N³-{6-[methyl(propanoyl)amino]-4-[(1 S)-2-methyl-1-(pyridin-2-yl)propy

3,4-dihydropyrido[2,3-b]pyrazin-2-yl}-beta-alaninamide

or a pharmaceutically acceptable salt thereof.

Preferred BET-family bromodomain inhibitors described in US provisional patent application serial number 62/181281 , which was filed on 18^th June 2015, include:

N³-{4-[(1 f?)-2-methoxy-1-phenylethyl]-6-[methyl(propanoyl)amino]-3-oxo-3,4- dihydropyrido[2,3-b]pyrazin-2-yl}-N-methyl-beta-alaninamide;

N-{6-[methyl(methylcarbamoyl)amino]-3-oxo-4-[(1 S)-1-phenylpropyl]-3,4- dihydropyrido[2,3-b]pyrazin-2-yl}-beta-alanine; and

N-methyl-N³-{6-[methyl(propanoyl)amino]-4-[(1 S)-2-methyl-1-(pyridin-2-yl)propyl]-3-oxo-

3,4-dihydropyrido[2,3-b]pyrazin-2-yl}-beta-alaninamide

or a pharmaceutically acceptable salt thereof.

An especially preferred BET-family bromodomain inhibitor described in US provisional patent application serial number 62/181281 , which was filed on 18^th June 2015, is:

N-{6-[methyl(propanoyl)amino]-3-oxo-4-[(1S)-1-phenylpropyl]-3,4-dihydropyrido[2,3- b]pyrazin-2-yl}-beta-alanine

or a pharmaceutically acceptable salt thereof.

Still further BET -family bromodomain inhibitors suitable for use with the methods of the present invention include those described in US provisional patent application serial number 62/181281 , which was filed on 18^th June 2015 the disclosure of which is hereby incorporated by reference in its entirety for all purposes. This disclosure includes compounds of Formula II:

Formula II

harmaceutically acceptable salt thereof, wherein

R¹ is selected from the group consisting of:

(i) -C3-C₇cycloalkyl optionally substituted with one, two, three or four E;

(ii) 4 to 7 membered heterocyclyl optionally substituted with one, two, three or four E, which said 4 to 7 membered heterocyclyl comprises one or two heteroatoms independently selected for each occurrence from the group of N, O and S; and

R^1A is selected from the group consisting of

(i) -d-Cealkyl optionally substituted with one, two, three, four, five or six E;

(ii) -C3-C₇cycloalkyl optionally substituted with one, two, three, four or five E;

(iii) phenyl optionally substituted with one, two, three, four or five E;

R^{1 B} is selected from the group consisting of

(i) -H; and

R^{1 C} is selected from the group consisting of

(i) -H;

(ii) -CH3 optionally substituted with one, two, or three J;

(iii) -CH2CH3 optionally substituted with one, two, three, four or five J;

W is selected from the group consisting of:

R4A

(iii)

(vi) 4 to 7 membered heterocyclyl optionally substituted with one, two, three or four G, which said 4 to 7 membered heterocyclyl comprises one, two, three or four heteroatoms independently selected for each occurrence from the group consisting of N, O and

S;

Y is selected from the group consisting of: (i) -CH₂- optionally substituted with one or two J;

(ii) -(CH2)2- optionally substituted with one, two, three or four J;

(iii) -(CH2)3- optionally substituted with one, two, three, four, five or six J; and

(iv) -(CH₂)4- optionally substituted with one, two, three, four, five, six, seven or eight J;

R³ is selected from the group consisting of:

(i) -H;

(ii) -CH3 optionally substituted with one, two, or three J;

(iii) -CH2CH3 optionally substituted with one, two, three, four or five J;

(v) CH(CH₃)₂ optionally substituted with one, two, three, four, five, six or seven J; R^4A is selected from the group consisting of

(i) -H;

(ii) -d-Cealkyl optionally substituted with one, two, three or four G;

(iv) -C(0)Ci-C6alkyl optionally substituted with one, two, three or four G;

(v) -C(0)OCi-C6alkyl optionally substituted with one, two, three or four

G;

(vi) -C(0)NH₂;

(viii) -C(0)N(Ci-C6alkyl)2 optionally substituted with one, two, three or four G;

(x) -NH(CrC3alkyl) optionally substituted with one, two, three or four

G;

(xi) -N(Ci-C3alkyl)2 optionally substituted with one, two, three or four

G; (xii) -NHC(0)CrC₃alkyl optionally substituted with one, two, three or four G;

(xiii) -N(Ci-C₃alkyl)C(0)Ci-C₃alkyl optionally substituted with one, two, three or four G;

(xvii) -S0₂NH(C C₃alkyl) optionally substituted with one, two, three or four G;

(xix) -C₃-C₇cycloalkyl optionally substituted with one, two, three or four

G;

(xx) phenyl optionally substituted with one, two, three or four G;

S;

elected from the group consisting of

(i) -H;

(ii) -d-Cealkyl optionally substituted with one, two, three or four G;

(iii) -C(0)Ci-C₆alkyl optionally substituted with one, two, three or four

G;

(iv) -C(0)OCi-C₆alkyl optionally substituted with one, two, three or four

G;

(v) -C(0)NH₂; (vi) -C(0)NH(Ci-C6alkyl) optionally substituted with one, two, three or four G;

(vii) -C(0)N(Ci-C6alkyl)2 optionally substituted with one, two, three or four G;

(viii) -C(0)NHSC>2Ci-C3alkyl optionally substituted with one, two, three or four G;

(ix) -C3-C₇cycloalkyl optionally substituted with one, two, three or four

G;

(x) phenyl optionally substituted with one, two, three or four G;

S;

elected from the group consisting of

(i) -H;

(ii) -d-Cealkyl optionally substituted with one, two, three or four G;

(iii) -C3-C₇cycloalkyl optionally substituted with one, two, three or four

G;

(iv) phenyl optionally substituted with one, two, three or four G;

(vi) 5 to 6 membered heteroaryl optionally substituted with one, two, three or four G, which said 5 to 6 membered heteroaryl ring comprises one, two, three or four heteroatoms independently selected for each occurrence from the group consisting of N, O and R¹⁰ is independently selected for each occurrence from the group consisting of - H, -F, -CI, -OH, -CN, -CH₃, -CH₂CH₃, -CH₂F, -CHF₂, -CF₃, -CF₂CF₃, -CH₂OH, - OCH₃, -OCH₂F, -OCHF₂, -OCF₃, -SCH₃, -SCH₂F, -SCHF₂, -SCF₃ -NH₂, - NH(CH₃), and -N(CH₃)₂;

E is independently selected for each occurrence from the group consisting of:

(i) -OH;

(ii) -CN;

(iii) -C0₂H;

(iv) -C(0)H;

(v) halo;

(vi) -Ci-C₃alkyl optionally substituted with one, two, three or four J;

(vii) -Ci-C₃alkylC0₂H which -Ci-C₃alkyl is optionally substituted with one, two, three or four J;

(x) -OCrC₃alkyl, optionally substituted with one, two, three or four J;

(xiii) -SCrC₃alkyl, optionally substituted with one, two, three or four J;

(xvi) -C(0)Ci-C₃alkyl, optionally substituted with one, two, three or four J;

(xviii) -NH₂;

(xix) -NH(Ci-C₃alkyl) optionally substituted with one, two, three or four J;

(xx) -N(Ci-C₃alkyl)₂ which -Ci-C₃alkyl is, independently for each occurrence, optionally substituted with one, two, three or four J;

(xxi) -C(0)NH₂;

(xxii) -C(0)NHCrC₃alkyl, optionally substituted with one, two, three or four J; (xxiii) -C(0)N(CrC3alkyl)₂, which -d-dalkyl is, independently for each occurrence, optionally substituted with one, two, three or four J;

(xxiv) -NHC(0)Ci-C3alkyl, optionally substituted with one, two, three or four J;

(xxv) -SC>2(Ci-C3alkyl), optionally substituted with one, two, three or four J; (xxvi) -SC>2NH(Ci-C3alkyl), optionally substituted with one, two, three or four J;

(xxvii) -NHSC>2(Ci-C3alkyl), optionally substituted with one, two, three or four J; and

(xxviii) phenyl optionally substituted with one, two, three, or four J;

G is independently selected for each occurrence from the group consisting of (i) -OH

(ii) -CN ;

(iv) -C(0)H;

(v) halo;

(vi) -Ci-C3alkyl, optionally substituted with one, two, three or four J;-

(vii) -C C3alkylCC>2H, which -d-dalkyl is optionally substituted with one, two, three or four J;

(viii) -Ci-C3alkylC3-C6cycloalkyl optionally substituted with one, two, three, four, five or six J;

(ix) -OC C3alkyl, optionally substituted with one, two, three or four J;

(x) -OCi-C3alkylC3-C6cycloalkyl optionally substituted with one, two, three, four, five or six J;

(xi) -SCrC3alkyl, optionally substituted with one, two, three or four J;

(xii) -SCi-C3alkylC3-C6cycloalkyl optionally substituted with one, two, three, four, five or six J;

(xiii) -C(0)Ci-C3alkyl, optionally substituted with one, two, three or four J;

(xiv) -C(0)OCi-C3alkyl, optionally substituted with one, two, three or four J;

(xv) -IMH2;

(xvi) -NH(Ci-C3alkyl), optionally substituted with one, two, three or four J ;

(xvii) -N(CrC3alkyl)₂, which -d-dalkyl is, independently for each occurrence, optionally substituted with one, two, three or four J;

(xviii) -C(0)NH₂;

(xix) -C(0)NHCrC3alkyl, optionally substituted with one, two, three or four J;

(xx) -C(0)N(Ci-C3alkyl)2, which -d-dalkyl is, independently for each

occurrence, optionally substituted with one, two, three or four J; (xxi) -NHC(0)CrC₃alkyl, optionally substituted with one, two, three or four J;

(xxii) -SC>₂(Ci-C₃alkyl), optionally substituted with one, two, three or four J;

(xxiii) -S0₂NH(Ci-C₃alkyl), optionally substituted with one, two, three or four J; and

J is independently selected for each occurrence from the group consisting of -H, -F, -CI, -OH, -CN, -CH₃, -CH₂CH₃, -CH₂F, -CHF₂, -CF₃, -CF₂CF₃, -CH₂OH, - OCH3, -OCH2F, -OCHF2, -OCF3, -SCH3, -SCH2F, -SCHF2, -SCF3 -NH₂, - NH(CH₃), and -N(CH₃)₂.

A preferred Embodiment of these compounds of Formula II is disclosed in US provisional patent application serial number 62/181281 , where:

R^{1 C} is -H;

Y is selected from the group consisting of -CH₂- and -CH₂CH₂-;

R³ is -H;

R¹⁰ is -H, or a pharmaceutically acceptable salt thereof.

Specific examples of BET-family bromodomain inhibitors, including their methods of preparation, of Formula II described in US provisional patent application serial number 62/181281 , which was filed on 18^th June 2015 include: 6-(3,5-dimethyl-1 ,2-oxazol-4-yl)-4-[(1 S)-1-(2-methoxyphenyl)ethyl]-2-{[2-(morpholin-4- yl)ethyl]amino}pyrido[2,3-b]pyrazin-3(4H)-one;

6-(3,5-dimethyl-1 ,2-oxazol-4-yl)-4-(2-ethoxybenzyl)-2-{[2-(morpholin-4- yl)ethyl]amino}pyrido[2,3-b]pyrazin-3(4H)-one;

4-benzyl-6-(3,5-dimethyl-1 ,2-oxazol-4-yl)-2-{[2-(morpholin-4-yl)ethyl]amino}pyrido[2,3- b]pyrazin-3(4H)-one;

6-(3,5-dimethyl-1 ,2-oxazol-4-yl)-2-{[2-(morpholin-4-yl)ethyl]amino}-4-[(1 )-1- phenylpropyl]pyrido[2,3-b]pyrazin-3(4H)-one;

6-(3,5-dimethyl-1 ,2-oxazol-4-yl)-2-{[2-(morpholin-4-yl)ethyl]amino}-4-[(1 S)-1- phenylpropyl]pyrido[2,3-b]pyrazin-3(4H)-one;

6-(3,5-dimethyl-1 ,2-oxazol-4-yl)-2-{[2-(morpholin-4-yl)ethyl]amino}-4-[(1 S)-1- phenylethyl]pyrido[2,3-b]pyrazin-3(4H)-one;

N-{6-(3,5-dimethyl-1 ,2-oxazol-4-yl)-3-oxo-4-[(1 S)-1-phenylethyl]-3,4-dihydropyrido[2,3- b]pyrazin-2-yl}glycine;

N-{6-(3,5-dimethyl-1 ,2-oxazol-4-yl)-3-oxo-4-[(1 S)-1-phenylethyl]-3,4-dihydropyrido[2,3- b]pyrazin-2-yl}-beta-alanine;

N-{6-(3,5-dimethyl-1 ,2-oxazol-4-yl)-4-[(1 S)-1-(2-methoxyphenyl)ethyl]-3-oxo-3,4- dihydropyrido[2,3-b]pyrazin-2-yl}-beta-alanine;

N-{6-(3,5-dimethyl-1 ,2-oxazol-4-yl)-3-oxo-4-[(1 S)-1-phenylpropyl]-3,4-dihydropyrido[2,3- b]pyrazin-2-yl}-beta-alanine;

N-{6-(3,5-dimethyl-1 ,2-oxazol-4-yl)-3-oxo-4-[(1 S)-1-(pyrimidin-2-yl)propyl]-3,4- dihydropyrido[2,3-b]pyrazin-2-yl}-beta-alanine;

N-{6-(3,5-dimethyl-1 ,2-oxazol-4-yl)-4-[(2S)-1-methoxybutan-2-yl]-3-oxo-3,4- dihydropyrido[2,3-b]pyrazin-2-yl}-beta-alanine;

N-{6-(3,5-dimethyl-1 ,2-oxazol-4-yl)-4-[(2 )-1-methoxybutan-2-yl]-3-oxo-3,4- dihydropyrido[2,3-b]pyrazin-2-yl}-beta-alanine;

N-[4-(1 ,3-dimethoxypropan-2-yl)-6-(3,5-dimethyl-1 ,2-oxazol-4-yl)-3-oxo-3,4- dihydropyrido[2,3-b]pyrazin-2-yl]-beta-alanine;

N-[6-(3,5-dimethyl-1 ,2-oxazol-4-yl)-3-oxo-4-(tetrahydro-2H-pyran-4-yl)-3,4- dihydropyrido[2,3-b]pyrazin-2-yl]-beta-alanine;

N³-{6-(3,5-dimethyl-1 ,2-oxazol-4-yl)-3-oxo-4-[(1 S)-1-phenylethyl]-3,4-dihydropyrido[2,3- b]pyrazin-2-yl}-N-(methylsulfonyl)-beta-alaninamide;

6-(3,5-dimethyl-1 ,2-oxazol-4-yl)-4-[(1 S)-1-phenylethyl]-2-{[2-(1 H-tetrazol-5- yl)ethyl]amino}pyrido[2,3-b]pyrazin-3(4H)-one; N-{6-(3,5-dimethyl-1 ,2-oxazol-4-yl)-3-oxo-4-[(1 S)-1-phenylbutyl]-3,4-dihydropyrido[2,3- b]pyrazin-2-yl}-beta-alanine;

N-{6-(3,5-dimethyl-1 ,2-oxazol-4-yl)-4-[(1 S)-2-methyl-1-phenylpropyl]-3-oxo-3,4- dihydropyrido[2,3-b]pyrazin-2-yl}-beta-alanine;

N-{6-(3,5-dimethyl-1 ,2-oxazol-4-yl)-4-[(1 S)-2-methyl-1-(pyridin-2-yl)propyl]-3-oxo-3,4- dihydropyrido[2,3-b]pyrazin-2-yl}-beta-alanine;

N-{4-[(1 S)-1-cyclohexylethyl]-6-(3,5-dimethyl-1 ,2-oxazol-4-yl)-3-oxo-3,4- dihydropyrido[2,3-b]pyrazin-2-yl}-beta-alanine;

N-[6-(3,5-dimethyl-1 ,2-oxazol-4-yl)-3-oxo-4-(pentan-3-yl)-3,4-dihydropyrido[2,3- b]pyrazin-2-yl]-beta-alanine; and

N²-{6-(3,5-dimethyl-1 ,2-oxazol-4-yl)-4-[(1 S)-1-(2-methoxyphenyl)ethyl]-3-oxo-3,4- dihydropyrido[2,3-b]pyrazin-2-yl}-N-methylglycinamide

or a pharmaceutically acceptable salt thereof.

A preferred BET-family bromodomain inhibitor of Formula II described in US provisional patent application serial number 62/181281 , which was filed on 18^th June 2015, is:

N-{6-(3,5-dimethyl-1 ,2-oxazol-4-yl)-4-[(2 )-1-methoxybutan-2-yl]-3-oxo-3,4- dihydropyrido[2,3-b]pyrazin-2-yl}-beta-alanine

or a pharmaceutically acceptable salt thereof.

As used herein, unless otherwise noted, "alkyl" whether used alone or as part of a substituent group refers to a saturated straight or branched hydrocarbon chain (ie a substituent obtained from a hydrocarbon by removal of a hydrogen) having from one to twenty carbon atoms or any number within this range, for example, from one to six carbon atoms, from one to four carbon atoms or from one to three carbon atoms. Designated numbers of carbon atoms (e.g. Ci-β) shall refer independently to the number of carbon atoms in an alkyl moiety or to the alkyl portion of a larger alkyl-containing substituent. Examples of alkyl groups include, but are not limited to, methyl, ethyl, n- propyl, /^'so-propyl, n-butyl, sec-butyl, /^'so-butyl, te/f-butyl, pentyl, isoamyl, hexyl and the like. Where so indicated, alkyl groups can be optionally substituted. In substituent groups with multiple alkyl groups such as N(Ci-C₆alkyl)₂, the alkyl groups may be the same or different.

As used herein, unless otherwise noted, "alkoxy" refers to groups of formula - Oalkyl, wherein "alkyl" is as defined herein. Designated numbers of carbon atoms (e.g. -OC1-C6) shall refer independently to the number of carbon atoms in the alkyl moiety of the alkoxy group, for example, but not limited to, from one to six carbon atoms or from one to three carbon atoms. Examples of alkoxy groups include, but are not limited to, methoxy, ethoxy, n-propoxy, /^'so-propoxy, n-butoxy, sec-butoxy, /so-butoxy, te/f-butoxy, and the like. Where so indicated, alkoxy groups can be optionally substituted.

As used herein, unless otherwise noted, "aryl" whether used alone or part of another group refers to a carbocyclic fully unsaturated or partially unsaturated single or fused ring system. If the rings are fused, one of the rings must be fully unsaturated or partially unsaturated and the fused ring(s) may be fully saturated, partially unsaturated or fully unsaturated. The aryl group may be optionally substituted as defined herein. The term "aryl" embraces aromatic radicals such as phenyl, naphthyl, tetrahydronaphthyl, indanyl, biphenyl, benzo[b][1 ,4]oxazin-3(4H)-onyl, 2,3-dihydro-1/-/ indenyl and 1 ,2,3,4- tetrahydronaphthalenyl.

As used herein, unless otherwise noted, "cycloalkyl" whether used alone or as part of another group, refers to a fully saturated hydrocarbon ring having from three to fourteen ring carbon atoms, for example, from four to seven; or from three to seven; or from three to six; or from three to five ring carbon atoms. Cycloalkyl groups can be monocyclic (e.g., cyclohexyl) or polycyclic (e.g., containing fused, bridged, and/or spiro ring systems), wherein the carbon atoms are located inside or outside of the ring system. Any suitable ring position of the cycloalkyl group can be covalently linked to the defined chemical structure. Where so indicated, cycloalkyl rings can be optionally substituted. Examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctanyl, decalinyl. The term "cycloalkyl" also includes carbocyclic rings which are bicyclic hydrocarbon rings, non- limiting examples of which include, bicyclo-[2.1.1]hexanyl, bicyclo[2.2.1]heptanyl, bicyclo[3.1.1]heptanyl, 1 ,3-dimethyl[2.2.1]heptan-2-yl, bicyclo[2.2.2]octanyl, and bicyclo[3.3.3]undecanyl.

As used herein, unless otherwise noted, the terms "haloalkyl" and "haloalkoxy" are intended to include both branched and straight-chain saturated aliphatic "alkyl" or "alkoxy" groups respectively, wherein "alkyl" and "alkoxy" are as defined herein, having the specified number of carbon atoms and in which at least one hydrogen is replaced with a halogen atom. As used herein, the term "halogen atom" refers to F, CI, Br and I. Haloalkyl groups include perhaloalkyl groups, wherein all hydrogens of an alkyl group have been replaced with halogens (e.g., -CF3, -CF2CF3). In certain embodiments in which two or more hydrogen atoms are replaced by halogen atoms, the halogen atoms can be the same (e.g. , CHF₂, -CF₃) or different (e.g. , CF₂CI). Where so indicated, haloalkyl or haloalkoxy groups can optionally be substituted with one or more substituents in addition to halogen. Examples of haloalkyl groups include, but are not limited to, fluoromethyl, dichloroethyl, trifluoromethyl, trichloromethyl, pentafluoroethyl, and pentachloroethyl groups.

As used herein, unless otherwise noted, the terms "heterocyclyl" and "heterocycloalkyl" are used interchangeably and, whether used alone or as part of another group, are defined herein as referring to a group having one or more rings (e.g., 1 , 2 or 3 rings) and having from 3 to 1 1 ring atoms (e.g. 3 to 6 ring atoms, 4 to 7 ring atoms, 4 to 5 ring atoms) wherein at least one ring atom, alternatively 1 to 5 ring atoms, alternatively 1 to 4 ring atoms, alternatively 1 to 3 ring atoms, alternatively one ring atom, alternatively two ring atoms, is a heteroatom, independently selected, unless indicated otherwise, from the group consisting of nitrogen (N), oxygen (O), and sulfur (S), and wherein the ring that includes the heteroatom is fully saturated. Exemplary heterocyclyl groups have from 3 to 1 1 ring atoms, alternatively 4 to 7 ring atoms, alternatively 4 to 5 ring atoms, alternatively 3 to 6 ring atoms, of which, where chemically possible, from 1 to 5, alternatively 1 to 4, alternatively 1 to 3, alternatively 4, alternatively 3, alternatively 2, alternatively 1 ring atom, is a heteroatoms independently selected in each instance from, unless indicated otherwise, the group consisting of nitrogen (N), oxygen (O), or sulfur (S). In a group that has a heterocyclyl substituent, unless otherwise stated, the ring atom of the heterocyclyl substituent that is bound to the group may be one of the heteroatoms, or it may be a ring carbon atom, where the ring carbon atom may be in the same ring as the heteroatom(s), or the ring carbon may be in a different ring from the heteroatom(s). Where so indicated, the heterocyclyl substituent can be optionally further substituted with one or more group(s) or substituent(s), which group(s) or substituent(s) may be bound to the heteroatom(s) or may be bound to the ring carbon atom, where the ring carbon atom may be in the same ring as the at least one heteroatom or where the ring carbon atom may be in a different ring rom the heteroatom(s). Examples of monocyclic heterocyclyl groups include, but are not limited to, oxetanyl, diazirinyl, aziridinyl, urazolyl, azetidinyl, pyrazolidinyl, imidazolidinyl, oxazolidinyl, isoxazolinyl, isoxazolyl, thiazolidinyl, isothiazolyl, isothiazolinyl oxathiazolidinonyl, oxazolidinonyl, hydantoinyl, tetrahydrofuranyl, pyrrolidinyl, morpholinyl, piperazinyl, piperidinyl, dihydropyranyl, tetrahydropyranyl, piperidin-2-onyl (valerolactam), 2,3,4,5-tetrahydro-1 /-/-azepinyl, 2, 3-dihydro-1 /-/-indole, and 1 ,2,3,4-tetrahydro-quinoline.

As used herein, unless otherwise noted, the term "heteroaryl" whether used alone or as part of another group, is defined herein as a single or fused ring system having from five to eleven ring atoms (e.g. from five to ten ring atoms of from five to six ring atoms) wherein at least one ring atom, alternatively 2 ring atoms, alternatively 3 ring atoms, alternatively 4 ring atoms, in at least one ring is a heteroatom independently selected in each instance from, unless otherwise indicated, the group consisting of nitrogen (N), oxygen (O), and sulfur (S), and wherein further at least one of the rings comprising a heteroatom is fully unsaturated or partially unsaturated. In heteroaryl groups that include 2 or more fused rings, additional rings may bear one or more heteroatoms, may be a carbocycle (e.g., 6,7-Dihydro-5/-/-cyclopentapyrimidine) or may be aryl (e.g., benzofuranyl, benzo-thiophenyl, indolyl, indolinyl, tetrahydroquinolinyl, chromanyl, 1 ,4-dioxochromanyl). In a group that has a heteroaryl substituent, unless otherwise indicated, the ring atom of the heteroaryl substituent that is bound to the group may be the at least one heteroatom, or it may be a ring carbon atom, where the ring carbon atom may be in the same ring as the at least one heteroatom or where the ring carbon may be in a different ring from the at least one heteroatom. Where so indicated, heteroaryl groups can be substituted. If the heteroaryl substituent is substituted with a group or substituent, the group or substituent may be bound to the heteroatom, or it may be bound to a ring carbon atom, where the ring carbon atom may be in the same ring as the heteroatom(s), or where the ring carbon atom may be in a different ring from the heteroatom(s). Examples of monocyclic heteroaryl rings include, but are not limited to, 1 ,2,3,4-tetrazolyl, [1 ,2,3]triazolyl, [1 ,2,4]triazolyl, triazinyl, thiazol- 2-yl, thiazol-4-yl, imidazol-1-yl, 1/-/-imidazol-2-yl, 1/-/-imidazol-4-yl, oxazolyl, isoxazolin- 5-yl, furan-2-yl, furan-3-yl, thiophen-2-yl, thiophen-4-yl, pyrimidin-2-yl, pyrimidin-4-yl, pyrimidin-5-yl, pyridazinyl, pyrazinyl, pyridin-2-yl, pyridin-3-yl, and pyridin-4-yl pyridinyl. Examples of heteroaryl rings containing 2 or more fused rings include, but are not limited to, benzofuranyl, benzothiophenyl, benzoxazolyl, benzthiazolyl, benztriazolyl, cinnolinyl, naphthyridinyl, benzimidazolyl, aza-indolyl, aza-benzimidazolyl, phenanthridinyl, 7/-/-purinyl, 9/-/-purinyl, 5/-/-pyrrolo[3,2-c]pyrimidinyl, 7/-/-pyrrolo[2,3- cflpyrimidinyl, pyrido[2,3-d]pyrimidinyl, 2-phenylbenzo[d]thiazolyl, 1 - -indolyl, 4,5,6,7- tetrahydro-1-/-/-indolyl, quinoxalinyl, 5-methylquinoxalinyl, quinazolinyl, quinolinyl, and isoquinolinyl. The term "heteroaryl" also includes pyridyl N-oxides and groups containing a pyridine N-oxide ring. As used herein, unless otherwise stated, the term "amino" refers to -NH₂.

As used herein, unless otherwise stated, the term "alkylamino" refers to - N(H)alkyl, the term "alkyl" having already been defined herein. Examples of alkylamino substituents include, but are not limited to, methylamino, ethylamino, and propylamino.

As used herein, unless otherwise stated, the term "dialkylamino" refers to -

N(alkyl)₂ where the two alkyls may be the same or different and where the term "alkyl" has already been defined herein. Examples of dialkylamino substituents include, but are not limited to, dimethylamino, diethylamino, ethylmethylamino, and dipropylamino.

As used herein, unless otherwise stated, the term "amido" refers to -C(=0)NH₂. As used herein, unless otherwise stated, the term "halogen" or "halogen atom" refers to the group consisting of fluorine (which may be depicted as -F), chlorine (which may be depicted as -CI), bromine (which may be depicted as -Br), or iodine (which may be depicted as -I).

As used herein, unless otherwise stated, the terms "hydroxy" and "hydroxyl" are used interchangeably and as used herein mean an -OH group. As used herein, unless otherwise noted, the terms "hydroxyalkyl" and "hydroxyalkoxy" are intended to include both branched and straight-chain saturated aliphatic "alkyl" or "alkoxy" groups respectively, wherein "alkyl" and "alkoxy" are as defined herein, having the specified number of carbon atoms and in which at least one hydrogen is replaced with a -OH group. Where so indicated, hydroxyalkyl and hydroxyalkoxy groups can optionally be substituted with one or more substituents in addition to -OH. Examples of hydroxyalkyl groups include, but are not limited to, CH₂OH, CH₂CH₂OH, CH₂(OH)CH₂OH.

As used herein, unless otherwise stated, the term "oxo" =0.

As used herein, unless otherwise stated, the term "carbonyl" refers to C=0.

As used herein, unless otherwise stated, the term "carboxy" refers to -C0₂H.

As used herein, unless otherwise stated, the term sulfonyl refers to -S0₂-.

As used herein, the term "substituted" is used throughout the specification. The term "substituted" is defined herein as a moiety, whether acyclic or cyclic, which has one or more (e.g. 1-10) hydrogen atoms replaced by a substituent as defined herein below. Substituents include those that are capable of replacing one or two hydrogen atoms of a single moiety at a time, and also those that can replace two hydrogen atoms on two adjacent carbons to form said substituent. For example, substituents that replace single hydrogen atoms include, but are not limited to, halogen, hydroxy, and the like. A two hydrogen atom replacement includes, but is not limited to, carbonyl, oximino, and the like. Substituents that replace two hydrogen atoms from adjacent carbon atoms include, but are not limited to, epoxy, and the like. When a moiety is described as "substituted" any number of its hydrogen atoms can be replaced, as described above. For example, difluoromethyl is a substituted Ci alkyl; trifluoromethyl is a substituted Ci alkyl; 4-hydroxyphenyl is a substituted aryl ring; (N,N-dimethyl-5- amino)octanyl is a substituted Cs alkyl; 3-guanidinopropyl is a substituted C3 alkyl; and 2-carboxy-3-fluoropyridinyl is a substituted heteroaryl.

A multi-moiety substituent is bound through the atom indicated by To illustrate this the term "-OCrCahydroxyalkyl" is an Od-Caalkyl group substituted by a hydroxy group. Further, any carbon number pre-fix attached to a multi-moiety substituent only applies to the moiety it immediately precedes. To illustrate, the term "cycloalkyl(Ci-C- alky contains two moieties: alkyl and cycloalkyl. The (C₁-C4) pre-fix on the cycloalkyl(Ci-C4)alkyl means that the alkyl moiety of the alkylcycloalkyl contains from 1 to 4 carbon atoms, the (C₁-C4) pre-fix does not describe the cycloalkyl moiety.

If a group of substituents are collectively described as being optionally substituted by one or more of a list of substituents, the group may include (1) unsubstitutable substituents, (2) substitutable substituents that are not substituted by the optional substituents, and / or (3) substitutable substituents that are substituted by one or more of the optional substituents.

If a substituent is described such that it "may be substituted" or as being "optionally substituted" with up to a particular number of non-hydrogen substituents, that substituent may be either (1) not substituted; or (2) substituted by up to that particular number of non-hydrogen substituents or by up to the maximum number of substitutable positions on the substituents, whichever is less. Thus, for example, if a substituent is described as a heteroaryl optionally substituted with one, two or three substituents, then any heteroaryl with less than three substitutable positions would be optionally substituted by up to only as many non-hydrogen substituents as the heteroaryl has substitutable positions. To illustrate, tetrazolyl (which has only one substitutable position) would be optionally substituted with up to one non-hydrogen substituent.

At various places in the present specification, substituents of compounds are disclosed in groups or in ranges. It is specifically intended that the description include each and every individual sub-combination of the members of such groups and ranges. For example, the term "C _-6 alkyl" is specifically intended to individually disclose Ci , C2, C3, C4, C5, C6, C1-C6, C1-C5, C1-C4, C1-C3, C1-C2, C2-C6, C2-C5, C2-C4, C2-C3, C3-C6, C3- C5, C3-C4, C4-C6, C4-C5, and C5-C6 alkyl. For example, the term "Ci_-3 alkyl" is specifically intended to individually disclose Ci , C2, C3, C1-C3, C1-C2, and C2-C3 alkyl.

Still further examples of BET-family bromodomain inhibitors for use with the present invention include those compounds generically or specifically disclosed in the following patent application publications: WO2015/004075; W02015/004534 WO2015/004533; WO2015/002754; US2015011540; WO2014/206150; WO14206345 WO2014/202578; WO2014/191894; WO2014/191906; WO2014/19191 1 US2014349990; US2014336190; WO2014/182929; WO2014/173241 ; EP2792355 WO2014/170350; US2014296246; WO2014/160873; US2014296229 WO2014/154760; US2014296230; WO2014/154762; WO2014/152029 US2014275030; WO2014/143768; W02014/145051 ; US2014275026; US2014275079

WO2014/139324 WO2014/134232; WO2014/134267; WO2014/128070 WO2014/128067 WO2014/12811 1 ; WO2014/128655; WO2014/095774 WO2014/095775 US2014162971 ; CN103833671 ; US2014140956; US2014142102 US2014142098; WO2014/076237; US2014135336; WO2014/048945 WO2014/026997 US2013331382; WO2013/184878 WO2013/184876 WO2013/155695 US2013281450; WO2013/158952 WO2013/097052 WO2013/097601 US2013079335; WO2013/033268 WO2013/033269 WO2013/030150 WO2013/027168; GB2011 14103; WO2013/024104 WO2012/151512 WO2012/150234; WO2012/143413; WO2012/143416 WO2012/143415 WO2012/1 16170; WO2012/075383; WO2012/075456 WO201 1/161031 GB200919426; WO2011/054844; WO201 1/054845 WO201 1/054553 WO201 1/054843; WO201 1/054848; WO201 1/054846 WO2009/084693 EP2239264; US2010286127 or pharmaceutically acceptable salts thereof, all of which are incorporated by reference herein in their entirety for all purposes.

In one embodiment of the present invention the BET-family bromodomain inhibitor is l-BET-762, or a pharmaceutically acceptable salt thereof.

In one embodiment of the present invention the BET-family bromodomain inhibitor is JQ-1 , or a pharmaceutically acceptable salt thereof.

In one embodiment of the present invention the BET-family bromodomain inhibitor is JQ-1 (+), or a pharmaceutically acceptable salt thereof. In one embodiment of the present invention BET-family bromodomain inhibitor is CPI-203, or a pharmaceutically acceptable salt thereof.

In one embodiment of the present invention the BET-family bromodomain inhibitor is OTX-015, or a pharmaceutically acceptable salt thereof.

In one embodiment of the present invention the BET-family bromodomain inhibitor is GW-841819X, or a pharmaceutically acceptable salt thereof.

In one embodiment of the present invention the BET-family bromodomain inhibitor is CP-0610, or a pharmaceutically acceptable salt thereof.

In one embodiment of the present invention the BET-family bromodomain inhibitor is CPI-232, or a pharmaceutically acceptable salt thereof.

In one embodiment of the present invention the BET-family bromodomain inhibitor is BET-BAY-002, or a pharmaceutically acceptable salt thereof.

In one embodiment of the present invention the BET-family bromodomain inhibitor is RVX-208, or a pharmaceutically acceptable salt thereof.

In one embodiment of the present invention the BET-family bromodomain inhibitor is l-BET-726, or a pharmaceutically acceptable salt thereof.

In one embodiment of the present invention the BET-family bromodomain inhibitor is SRX-2523, or a pharmaceutically acceptable salt thereof.

In one embodiment of the present invention the BET-family bromodomain inhibitor is B S-988158, or a pharmaceutically acceptable salt thereof.

In In one embodiment of the present invention the BET-family bromodomain inhibitor is RVX-297, or a pharmaceutically acceptable salt thereof.

In one embodiment of the present invention the BET-family bromodomain inhibitor is RVS-2135, or a pharmaceutically acceptable salt thereof.

In one embodiment of the present invention the BET-family bromodomain inhibitor is ZEN-3365, or a pharmaceutically acceptable salt thereof.

In one embodiment of the present invention the BET-family bromodomain inhibitor is ZEN-31 18, or a pharmaceutically acceptable salt thereof.

In one embodiment of the present invention the BET-family bromodomain inhibitor is KM-601 , or a pharmaceutically acceptable salt thereof. In one embodiment of the present invention the BET-family bromodomain inhibitor is SF-2535, or a pharmaceutically acceptable salt thereof.

In one embodiment of the present invention the BET-family bromodomain inhibitor is AU-004, or a pharmaceutically acceptable salt thereof.

In one embodiment of the present invention the BET-family bromodomain inhibitor is ABBV-075, or a pharmaceutically acceptable salt thereof.

In one embodiment of the present invention the BET-family bromodomain inhibitor is TEN-010, or a pharmaceutically acceptable salt thereof.

In one embodiment of the present invention the BET-family bromodomain inhibitor is BAY1238097, or a pharmaceutically acceptable salt thereof.

In one embodiment of the present invention the BET-family bromodomain inhibitor is N-(2-hydroxy-3-methylquinolin-6-yl)piperidine-1-sulfonamide, or a pharmaceutically acceptable salt thereof.

In one embodiment of the present invention the BET-family bromodomain inhibitor is N-[4-(2,4-difluorophenoxy)-3-(6-methyl-7-oxo-6,7-dihydro-1 H-pyrrolo[2,3- c]pyridin- 4-yl)phenyl]ethanesulfonamide, or a pharmaceutically acceptable salt thereof.

In one embodiment of the present invention the BET-family bromodomain inhibitor is N-{6-[methyl(propanoyl)amino]-3-oxo-4-[(1S)-1-phenylpropyl]-3,4- dihydropyrido[2,3 b]pyrazin-2-yl}-beta-alanine, or a pharmaceutically acceptable salt thereof.

In a preferred embodiment of the present invention the BET-family bromodomain inhibitor is selected from the group consisting of:

I-BET762;

JQ-1 ;

JQ-1 (+);

CPI-267203;

OTX-015;

GW-841819X;

CP-0610;

CPI-232; BET-BAY-002;

l-BET-151 ;

RVX-208;

l-BET-726;

SRX-2523;

N-(2-hydroxy-3-methylquinolin-6-yl)piperidine-1-sulfonamide;

N-[4-(2,4-difluorophenoxy)-3-(6-methyl-7-oxo-6,7-dihydro-1 H-pyrrolo[2,3- c]pyridin- 4-yl)phenyl]ethanesulfonamide; and

N-{6-[methyl(propanoyl)amino]-3-oxo-4-[(1 S)-1-phenylpropyl]-3,4- dihydropyrido[2,3 b]pyrazin-2-yl}-beta-alanine

or a pharmaceutically acceptable salt thereof.

I-BET762;

JQ-1 ;

JQ-1 (+);

RVX-208;

l-BET-151 ;

N-(2-hydroxy-3-methylquinolin-6-yl)piperidine-1-sulfonamide;

or a pharmaceutically acceptable salt thereof.

Compounds useful for the present invention may be administered in an unsolvated or solvated forms. The term "solvate" as used herein means a physical association of a compound with one or more solvent molecules, whether organic or inorganic, including water ('hydrate'). When the solvent or water is tightly bound, the complex will have a well-defined stoichiometry independent of humidity. When, however, the solvent or water is weakly bound, as in channel solvates and hygroscopic compounds, the water/solvent content will be dependent on humidity and drying conditions. In such cases, non-stoichiometry will be the norm.

Compounds useful for the present invention may be used in the form of salts derived from inorganic or organic acids. Depending on the particular compound, a salt of the compound may be advantageous due to one or more of the salt's physical properties, such as enhanced pharmaceutical stability in differing temperatures and humidities, or a desirable solubility in water or oil. In some instances, a salt of a compound also may be used as an aid in the isolation, purification, and/or resolution of the compound.

Where a salt is intended to be administered to a patient (as opposed to, for example, being used in an in vitro context), the salt preferably is pharmaceutically acceptable. The term "pharmaceutically acceptable salt" refers to a salt prepared by combining a compound useful for the invention (e.g. a compound of Formula (I)) with an acid whose anion, or a base whose cation, is generally considered suitable for human consumption. Pharmaceutically acceptable salts are particularly useful as products of the methods of the present invention because of their greater aqueous solubility relative to the parent compound. For use in medicine, the salts of the compounds useful for this invention are non-toxic "pharmaceutically acceptable salts." Salts encompassed within the term "pharmaceutically acceptable salts" refer to non-toxic salts of the compounds of this invention which are generally prepared by reacting the free base with a suitable organic or inorganic acid.

When possible, suitable pharmaceutically acceptable acid addition salts of the compounds useful for the present invention include those derived from inorganic acids, such as hydrochloric, hydrobromic, hydrofluoric, boric, fluoroboric, phosphoric, metaphosphoric, nitric, carbonic, sulfonic, and sulfuric acids, and organic acids such as acetic, benzenesulfonic, benzoic, citric, ethanesulfonic, fumaric, gluconic, glycolic, isothionic, lactic, lactobionic, maleic, malic, methanesulfonic, trifluoromethanesulfonic, succinic, toluenesulfonic, tartaric, and trifluoroacetic acids. Suitable organic acids generally include but are not limited to aliphatic, cycloaliphatic, aromatic, araliphatic, heterocyclic, carboxylic, and sulfonic classes of organic acids.

Specific examples of suitable organic acids include but are not limited to acetate, trifluoroacetate, formate, propionate, succinate, glycolate, gluconate, digluconate, lactate, malate, tartaric acid, citrate, ascorbate, glucuronate, maleate, fumarate, pyruvate, aspartate, glutamate, benzoate, anthranilic acid, stearate, salicylate, p- hydroxybenzoate, phenylacetate, mandelate, embonate (pamoate), methanesulfonate, ethanesulfonate, benzenesulfonate, pantothenate, toluenesulfonate, 2- hydroxyethanesulfonate, sufanilate, cyclohexylaminosulfonate, algenic acid, .beta.- hydroxybutyric acid, galactarate, galacturonate, adipate, alginate, butyrate, camphorate, camphorsulfonate, cyclopentanepropionate, dodecylsulfate, glycoheptanoate, glycerophosphate, heptanoate, hexanoate, nicotinate, 2-naphthalesulfonate, oxalate, palmoate, pectinate, 3-phenylpropionate, picrate, pivalate, thiocyanate, and undecanoate.

Furthermore, where the compounds useful for the present invention carry an acidic moiety, suitable pharmaceutically acceptable salts thereof may include alkali metal salts, i.e., sodium or potassium salts; alkaline earth metal salts, e.g., calcium or magnesium salts; and salts formed with suitable organic ligands, e.g., quaternary ammonium salts. In another embodiment, base salts are formed from bases which form non-toxic salts, including aluminum, arginine, benzathine, choline, diethylamine, diolamine, glycine, lysine, meglumine, olamine, tromethamine and zinc salts.

Organic salts may be made from secondary, tertiary or quaternary amine salts, such as tromethamine, diethylamine, Ν,Ν'-benzylethylenediamine, chloroprocaine, choline, diethanolamine, ethylenediamine, meglumine (N-methylglucamine), and procaine. Basic nitrogen-containing groups may be quaternized with agents such as lower alkyl (C.sub.1-C.sub.6) halides (e.g., methyl, ethyl, propyl, and butyl chlorides, bromides, and iodides), dialkyl sulfates (i.e., dimethyl, diethyl, dibutyl, and diamyl sulfates), long chain halides (i.e., decyl, lauryl, myristyl, and stearyl chlorides, bromides, and iodides), arylalkyl halides (i.e., benzyl and phenethyl bromides), and others.

Compounds useful for the present invention may also be administered in the form of hemisalts of acids and bases, for example, hemisulphate and hemicalcium salts.

Compounds useful for the present invention may also be administered in the form of complexes. Complexes, such as clathrates, drug-host inclusion complexes wherein, in contrast to the aforementioned solvates, the drug and host are present in stoichiometric or non-stoichiometric amounts. Also included are complexes of the drug containing two or more organic and/or inorganic components which may be in stoichiometric or non-stoichiometric amounts. The resulting complexes may be ionised, partially ionised, or non-ionised. For a review of such complexes, see J Pharm Sci, 64 (8), 1269-1288 by Haleblian (August 1975). Compounds useful for the present invention may also be administered as a prodrug. The term "prodrug" as used herein, refers to a pharmacologically inactive derivative of a parent "drug" molecule that requires biotransformation (e.g., either spontaneous or enzymatic) within the target physiological system to release or convert the prodrug into the active drug. Prodrugs are designed to overcome problems associated with stability, toxicity, lack of specificity, or limited bioavailability. Exemplary prodrugs comprise an active drug molecule itself and a chemical masking group (e.g., a group that reversibly suppresses the activity of the drug). Some preferred prodrugs are variations or derivatives of compounds that have groups cleavable under metabolic conditions. Exemplary prodrugs become pharmaceutically active in vivo or in vitro when they undergo solvolysis under physiological conditions or undergo enzymatic degradation or other biochemical transformation (e.g., phosphorylation, hydrogenation, dehydrogenation, glycosylation). Prodrugs often offer advantages of solubility, tissue compatibility, or delayed release in the mammalian organism. (See e.g., Bundgard, Design of Prodrugs, pp. 7-9, 21- 24, Elsevier, Amsterdam (1985); and Silverman, The Organic Chemistry of Drug Design and Drug Action, pp. 352-401 , Academic Press, San Diego, CA (1992)). Common prodrugs include acid derivatives such as esters prepared by reaction of parent acids with a suitable alcohol (e.g., a lower alkanol), amides prepared by reaction of the parent acid compound with an amine, or basic groups reacted to form an acylated base derivative (e.g., a lower alkylamide).

Compounds useful for the present invention may also be administered as isotopically labelled derivatives of the BET-family bromodomain inhibitor wherein one or more atoms are replaced by atoms having the same atomic number, but an atomic mass or mass number different from the atomic mass or mass number usually found in nature. Examples of isotopes suitable for inclusion in the compounds useful for the present invention include isotopes of hydrogen, such as ²H and ³H, carbon, such as ¹¹C, ¹³C and ¹⁴C, chlorine, such as ³⁶CI, fluorine, such as ¹⁸F, iodine, such as ¹²³l and ¹²⁵l, nitrogen, such as ¹³N and ¹⁵N, oxygen, such as ¹⁵0, ¹⁷0 and ¹⁸0, phosphorus, such as ³²P, and sulphur, such as ³⁵S. Substitution with heavier isotopes such as deuterium, i.e. ²H, may afford certain therapeutic advantages resulting from greater metabolic stability, for example, increased in vivo half-life or reduced dosage requirements, and hence may be preferred in some circumstances. Substitution with positron emitting isotopes, such as ¹¹C, ¹⁸F, ¹⁵0 and ¹³N, can be useful in Positron Emission Topography (PET) studies for examining substrate receptor occupancy. Isotopically-labeled compounds can generally be prepared by conventional techniques known to those skilled in the art or by processes analogous to those described.

Pharmaceutical Dosage Forms

For the pharmaceutical methods of the present invention, BET-family bromodomain inhibitors, and pharmaceutically acceptable salts thereof, will usually be administered in a pharmaceutically acceptable composition.

Methods of formulation of such pharmaceutically acceptable compositions are well known in the art and are disclosed, for example, in Remington: The Science and Practice of Pharmacy, Mack Publishing Company, Easton, Pa., 21st Edition (2005), incorporated herein by reference in its entirety for all purposes.

Pharmaceutical compositions suitable for use in the present invention can be in the form of sterile, non-pyrogenic liquid solutions or suspensions, coated capsules, suppositories, lyophilized powders, transdermal patches or other forms known in the art.

Injectable preparations, for example, sterile injectable aqueous or oleaginous suspensions may be formulated according to the known art using suitable dispersing or wetting agents and suspending agents. The sterile injectable preparation may also be a sterile injectable solution, suspension or emulsion in a nontoxic parenterally acceptable diluent or solvent.

In addition, sterile, fixed oils are conventionally employed as a solvent or suspending medium. For this purpose any bland fixed oil may be employed including synthetic mono- or di-glycerides. In addition, fatty acids such as oleic acid find use in the preparation of injectables. The injectable formulations can be sterilized, for example, by filtration through a bacterial- retaining filter, or by incorporating sterilizing agents in the form of sterile solid compositions which can be dissolved or dispersed in sterile water or other sterile injectable medium prior to use.

Formulations are provided herein comprising crystalline forms of the pharmaceutically active agent for slow absorption from subcutaneous or intramuscular injection. Additionally, delayed absorption of a parenterally administered drug form may be accomplished by dissolving or suspending the compounds in an oil vehicle. Injectable depot forms are made by forming microencapsule matrices of the drug in biodegradable polymers such as polylactide-polyglycolide. Depending upon the ratio of drug to polymer and the nature of the particular polymer employed, the rate of drug release can be controlled. Examples of other biodegradable polymers include poly(orthoesters) and poly(anhydrides). Depot injectable formulations may also be prepared by entrapping the drug in liposomes or microemulsions, which are compatible with body tissues.

Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In such solid dosage forms, the active compound is mixed with at least one inert, pharmaceutically acceptable excipient or carrier such as sodium citrate or dicalcium phosphate and/or a) fillers or extenders such as starches, lactose, sucrose, glucose, mannitol, and silicic acid, b) binders such as, for example, carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidinone, sucrose, and acacia, c) humectants such as glycerol, d) disintegrating agents such as agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates, and sodium carbonate, e) solution retarding agents such as paraffin, f) absorption accelerators such as quaternary ammonium compounds, g) wetting agents such as, for example, acetyl alcohol and glycerol monostearate, h) absorbents such as kaolin and bentonite clay, and i) lubricants such as talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, and mixtures thereof. In the case of capsules, tablets and pills, the dosage form may also comprise buffering agents.

Solid compositions of a similar type may also be employed as fillers in soft and hard-filled gelatin capsules using such excipients ass lactose or milk sugar as well as high molecular weight polyethylene glycols and the like.

The solid dosage forms of tablets, capsules, pills, and granules can be prepared with coatings and shells such as enteric coatings and other coatings well known in the pharmaceutical formulating art. They may optionally contain opacifying agents and can also be of a composition that they release the active ingredient(s) only, or preferentially, in a certain part of the intestinal tract, optionally, in a delayed manner. Examples of embedding compositions that can be used include polymeric substances and waxes.

Alternative formulations include those where the pharmaceutically active agent is in micro-encapsulated form with one or more excipients as noted above. The solid dosage forms of tablets, capsules, pills, and granules can be prepared with coatings and shells such as enteric coatings, release controlling coatings and other coatings well known in the pharmaceutical formulating art. In such solid dosage forms the active compound may be admixed with at least one inert diluent such as sucrose, lactose or starch. Such dosage forms may also comprise, as is normal practice, additional substances other than inert diluents, e.g., tableting lubricants and other tableting aids such a magnesium stearate and microcrystalline cellulose. In the case of capsules, tablets and pills, the dosage forms may also comprise buffering agents. They may optionally contain opacifying agents and can also be of a composition that they release the active ingredient(s) only, or preferentially, in a certain part of the intestinal tract, optionally, in a delayed manner. Examples of embedding compositions that can be used include polymeric substances and waxes.

Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups and elixirs. In addition to the active compounds, the liquid dosage forms may contain inert diluents commonly used in the art such as, for example, water or other solvents, solubilizing agents and emulsifiers such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, EtOAc, benzyl alcohol, benzyl benzoate, propylene glycol, 1 ,3-butylene glycol, dimethylformamide, oils (in particular, cottonseed, groundnut, corn, germ, olive, castor, and sesame oils), glycerol, tetrahydrofurfuryl alcohol, polyethylene glycols and fatty acid esters of sorbitan, and mixtures thereof. Besides inert diluents, the oral compositions can also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.

Dosage forms for topical or transdermal administration of a compound of this invention include ointments, pastes, creams, lotions, gels, powders, solutions, sprays, inhalants or patches. The pharmaceutically active agent is admixed under sterile conditions with a pharmaceutically acceptable carrier and any needed preservatives or buffers as may be required. Ophthalmic formulations, ear drops, and the like are also contemplated.

Compositions may also be formulated for delivery as a liquid aerosol or inhalable dry powder. Liquid aerosol formulations may be nebulized predominantly into particle sizes that can be delivered to the terminal and respiratory bronchioles.

The phrase "pharmaceutically acceptable carrier" as used herein means a pharmaceutical ly-acceptable material, composition or vehicle, such as a liquid or solid filler, diluent, excipient, solvent or encapsulating material, involved in carrying or transporting the pharmaceutically active agent from one organ, or portion of the body, to another organ, or portion of the body. Each carrier must be "acceptable" in the sense of being compatible with the other ingredients of the formulation and not injurious to the patient. Some examples of materials which can serve as pharmaceutically-acceptable carriers include: (1) sugars, such as lactose, glucose and sucrose; (2) starches, such as corn starch and potato starch; (3) cellulose, and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; (4) powdered tragacanth; (5) malt; (6) gelatin; (7) talc; (8) excipients, such as cocoa butter and suppository waxes; (9) oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; (10) glycols, such as propylene glycol; (11) polyols, such as glycerin, sorbitol, mannitol and polyethylene glycol; (12) esters, such as ethyl oleate and ethyl laurate; (13) agar; (14) buffering agents, such as magnesium hydroxide and aluminum hydroxide; (15) alginic acid; (16) pyrogen-free water; (17) isotonic saline; (18) Ringer's solution; (19) ethyl alcohol; (20) phosphate buffer solutions; and (21) other non-toxic compatible substances employed in p harmaceutical formulations. A physiologically acceptable carrier should not cause significant irritation to an organism and does not abrogate the biological activity and properties of the pharmaceutically active agent.

An "excipient" refers to an inert substance added to a pharmacological composition to further facilitate administration of a pharmaceutically active agent. Examples of excipients include but are not limited to calcium carbonate, calcium phosphate, various sugars and types of starch, cellulose derivatives, gelatin, vegetable oils and polyethylene glycols.

Dosing and Administration

The compounds useful for the invention described herein may be administered to humans and other animals orally, parenterally, sublingually, by aerosolization or inhalation spray, rectally, intracisternally, intravaginally, intraperitoneally, bucally, intrathecally or topically in dosage unit formulations containing conventional nontoxic pharmaceutically acceptable carriers, adjuvants, and vehicles as desired. The term parenteral as used herein includes subcutaneous injection, intravenous injection, intramuscular injection, intrasternal injection, or infusion techniques. Topical administration may also involve the use of transdermal administration such as transdermal patches or ionophoresis devices.

Effective amounts of the compounds useful for the invention generally include any amount sufficient to detectably modulate BET-family bromodomain activity, or to alleviate symptoms associated with diseases associated with BET-family bromodomain activity or susceptible to BET-family bromodomain activity modulation. The specific dose of compound administered according to the methods of the invention to obtain therapeutic and/or prophylactic effect will vary depending upon the host treated and the particular mode of administration. It will be understood, however, that the specific dose level for any particular subject will depend upon a variety of factors including, for example, the specific compound administered, the route of administration, the condition being treated, and the individual being treated, including the age, body weight, general health, sex, diet, of the individual, the time of administration, route of administration, rate of excretion, drug combination, and the severity of the particular disease undergoing therapy. The therapeutically effective amount for a given situation can be readily determined by routine experimentation and is within the skill and judgment of the ordinary clinician. A typical daily dose (administered in single or divided doses) will contain a dosage level of from about 0.01 mg/kg to about 50-100 mg/kg of body weight of an active compound useful for the invention. Preferred daily doses generally will be from about 0.05 mg/kg to about 20 mg/kg and ideally from about 0.1 mg/kg to about 10 mg/kg. Factors such as clearance rate, half-life and maximum tolerated dose (MTD) can determine these using standard procedures. Furthermore, it will be understood by those skilled in the art that the dosing regimen may be adjusted to balance treatment efficacy whilst minimizing patient adverse events and thereby improving the safety profile. Such dosing adjustments would be made by taking into account routine parameters such as the half life of frataxin and drug exposure.

In certain embodiments, the present invention relates to any of the aforementioned embodiments, wherein the method comprises co-administering the BET-family bromodomain inhibitor, or pharmaceutically acceptable salt thereof, which a second pharmaceutically active ingredient, or pharmaceutically acceptable salt thereof.

As used herein, the terms "co-administration", "co-administered", "a combination of or "in combination with", refers to a combination of a compound of Formula (I) and one or more other pharmaceutically active ingredient, or a pharmaceutically acceptable salt thereof, includes the following:

a. simultaneous administration of such a combination of a BET-family bromodomain inhibitor and a further pharmaceutically active agent to a patient in need of treatment, when such components are formulated together into a single dosage form which releases said components at substantially the same time to said patient, b. substantially simultaneous administration of such a combination of a BET-family bromodomaininhibitor and a further pharmaceutically active agent to a patient in need of treatment, when such components are formulated apart from each other into separate dosage forms which are taken at substantially the same time by said patient, whereupon said components are released at substantially the same time to said patient,

c. sequential administration of such a combination of a BET-family bromodomain inhibitor and a further pharmaceutically active agent to a patient in need of treatment, when such components are formulated apart from each other into separate dosage forms which are taken at consecutive times by said patient with a significant time interval between each administration, whereupon said components are released at substantially different times to said patient; and, d. sequential administration of such a combination of a BET-family bromodomain inhibitor and a further pharmaceutically active agent to a patient in need of treatment, when such components are formulated together into a single dosage form which releases said components in a controlled manner.

In particular it is contemplated that the compounds useful for the present invention may be co-administered with one or more therapeutic agents selected from the group consisting of:

an antihypertensive agent. Examples of suitable anti-hypertensive agents include: alpha adrenergic blockers; beta adrenergic blockers; calcium channel blockers (e.g., diltiazem, verapamil, nifedipine and amlodipine); vasodilators (e.g., hydralazine), diruetics (e.g., chlorothiazide, hydrochlorothiazide, flumethiazide, hydroflumethiazide, bendroflumethiazide, methylchlorothiazide, trichloromethiazide, polythiazide, benzthiazide, ethacrynic acid tricrynafen, chlorthalidone, torsemide, furosemide, musolimine, bumetanide, triamtrenene, amiloride, spironolactone); renin inhibitors; ACE inhibitors (e.g., captopril, zofenopril, fosinopril, enalapril, ceranopril, cilazopril, delapril, pentopril, quinapril, ramipril, lisinopril); AT-1 receptor antagonists (e.g., losartan, irbesartan, valsartan); ET receptor antagonists (e.g., sitaxsentan, atrsentan and compounds disclosed in U.S. Patent Nos. 5,612,359 and 6,043,265); Dual ET/AII antagonist (e.g., compounds disclosed in WO 00/01389); neutral endopeptidase (NEP) inhibitors; vasopepsidase inhibitors (dual NEP-ACE inhibitors) (e.g., gemopatrilat and nitrates). An exemplary antianginal agent is ivabradine; a calcium channel blocker (L-type or T-type) including diltiazem, verapamil, nifedipine and amlodipine and mybefradil. Examples of suitable cardiac glycosides include digitalis and ouabain;

a diuretic. Examples of suitable diuretics include (a) loop diuretics such as furosemide (such as LASIX™), torsemide (such as DEMADEX™), bemetanide (such as BUMEX™), and ethacrynic acid (such as EDECRIN™); (b) thiazide- type diuretics such as chlorothiazide (such as DIURIL™, ESIDRIX™ or HYDRODIURIL™), hydrochlorothiazide (such as MICROZIDE™ or ORETIC™), benzthiazide, hydroflumethiazide (such as SALURON™), bendroflumethiazide, methychlorthiazide, polythiazide, trichlormethiazide, and indapamide (such as LOZOL™); (c) phthalimidine-type diuretics such as chlorthalidone (such as HYGROTON™), and metolazone (such as ZAROXOLYN™); (d) quinazoline- type diuretics such as quinethazone; and (e) potassium-sparing diuretics such as triamterene (such as DYRENIUM™), and amiloride (such as MIDAMOR™ or MODURETIC™). In another embodiment, a compound useful for the present invention may be co-administered with a loop diuretic. In still another embodiment, the loop diuretic is selected from furosemide and torsemide. In still another embodiment, one or more compounds useful for the present invention may be co-administered with furosemide. In still another embodiment, one or more compounds useful for the present invention may be co-administered with torsemide which may optionally be a controlled or modified release form of torsemide. The diuretic can be a thiazide-type diuretic selected from the group consisting of chlorothiazide and hydrochlorothiazide. In still another embodiment, one or more compounds useful for the invention may be co-administered with chlorothiazide. In still another embodiment, one or more compounds useful for the present invention may be co-administered with hydrochlorothiazide. In another embodiment, one or more compounds useful for the invention may be co-administered with a phthalimidine-type diuretic. In still another embodiment, the phthalimidine-type diuretic is chlorthalidone;

an anti-diabetic agent, particularly type 2 anti-diabetic agent. Examples of suitable anti-diabetic agents include (e.g. insulins, metfomin, DPPIV inhibitors, GLP-1 agonists, analogues and mimetics, SGLT1 and SGLT2 inhibitors) Suitable anti-diabetic agents include an acetyl-CoA carboxylase- (ACC) inhibitor such as those described in WO2009144554, WO2003072197, WO2009144555 and WO2008065508, a diacylglycerol O-acyltransferase 1 (DGAT-1) inhibitor, such as those described in WO09016462 or WO2010086820, AZD7687 or LCQ908, diacylglycerol O-acyltransferase 2 (DGAT-2) inhibitor, monoacylglycerol O-acyltransferase inhibitors, a PDE10 inhibitor, an AMPK activator, a sulfonylurea (e.g., acetohexamide, chlorpropamide, diabinese, glibenclamide, glipizide, glyburide, glimepiride, gliclazide, glipentide, gliquidone, glisolamide, tolazamide, and tolbutamide), a meglitinide, an a-amylase inhibitor (e.g., tendamistat, trestatin and AL-3688), an a-glucoside hydrolase inhibitor (e.g., acarbose), an a-glucosidase inhibitor (e.g., adiposine, camiglibose, emiglitate, miglitol, voglibose, pradimicin-Q, and salbostatin), a PPARv agonist (e.g., balaglitazone, ciglitazone, darglitazone, englitazone, isaglitazone, pioglitazone and rosiglitazone), a PPAR α/γ agonist (e.g., CLX-0940, GW-1536, GW-1929, GW-2433, KRP-297, L-796449, LR-90, MK-0767 and SB-219994), a biguanide (e.g., metformin), a glucagon-like peptide 1 (GLP-1) modulator such as an agonist (e.g., exendin-3 and exendin-4), liraglutide, albiglutide, exenatide (Byetta®), albiglutide, lixisenatide, dulaglutide, semaglutide, NN-9924,TTP-054, a protein tyrosine phosphatase-1 B (PTP-1 B) inhibitor (e.g., trodusquemine, hyrtiosal extract, and compounds disclosed by Zhang, S., et al., Drug Discovery Today, 12(9/10), 373-381 (2007)), SIRT-1 inhibitor (e.g., resveratrol, GSK2245840 or GSK184072), a dipeptidyl peptidease IV (DPP-IV) inhibitor (e.g., those in WO2005116014, sitagliptin, vildagliptin, alogliptin, dutogliptin, linagliptin and saxagliptin), an insulin secreatagogue, a fatty acid oxidation inhibitor, an A2 antagonist, a c-jun amino-terminal kinase (JNK) inhibitor, glucokinase activators (GKa) such as those described in WO2010103437, WO2010103438, WO2010013161 , WO2007122482, TTP-399, TTP-355, TTP-547, AZD1656, ARRY403, MK-0599, TAK-329, AZD5658 or GKM-001 , insulin, an insulin mimetic, a glycogen phosphorylase inhibitor (e.g. GSK1362885), a VPAC2 receptor agonist, SGLT2 inhibitors, such as those described in E.C. Chao et al. Nature Reviews Drug Discovery 9, 551-559 (July 2010) including dapagliflozin, canagliflozin, empagliflozin, tofogliflozin (CSG452), ASP-1941 , THR1474, TS- 071 , ISIS388626 and LX421 1 as well as those in WO2010023594, a glucagon receptor modulator such as those described in Demong, D.E. et al. Annual Reports in Medicinal Chemistry 2008, 43, 1 19-137, GPR119 modulators, particularly agonists, such as those described in WO2010140092, WO2010128425, WO2010128414, WO2010106457, Jones, R.M. et al. in Medicinal Chemistry 2009, 44, 149-170 (e.g. MBX-2982, GSK1292263, APD597 and PSN821), FGF21 derivatives or analogs such as those described in Kharitonenkov, A. et al. et al., Current Opinion in Investigational Drugs 2009, 10(4)359-364, TGR5 (also termed GPBAR1) receptor modulators, particularly agonists, such as those described in Zhong, M., Current Topics in Medicinal Chemistry, 2010, 10(4), 386-396 and INT777, GPR40 agonists, such as those described in Medina, J.C., Annual Reports in Medicinal Chemistry, 2008, 43, 75- 85, including but not limited to TAK-875, GPR120 modulators, particularly agonists, high affinity nicotinic acid receptor (HM74A) activators, and SGLT1 inhibitors, such as GSK1614235. A further representative listing of anti-diabetic agents that can be combined with the compounds of the present invention can be found, for example, at page 28, line 35 through page 30, line 19 of WO2011005611. Preferred anti-diabetic agents are metformin and DPP-IV inhibitors (e.g., sitagliptin, vildagliptin, alogliptin, dutogliptin, linagliptin and saxagliptin). Other antidiabetic agents could include inhibitors or modulators of carnitine palmitoyl transferase enzymes, inhibitors of fructose 1 ,6-diphosphatase, inhibitors of aldose reductase, mineralocorticoid receptor inhibitors, inhibitors of TORC2, inhibitors of CCR2 and/or CCR5, inhibitors of PKC isoforms (e.g. PKCa, ΡΚΟβ, PKCy), inhibitors of fatty acid synthetase, inhibitors of serine palmitoyl transferase, modulators of GPR81 , GPR39, GPR43, GPR41 , GPR105, Kv1.3, retinol binding protein 4, glucocorticoid receptor, somatostain receptors (e.g. SSTR1 , SSTR2, SSTR3 and SSTR5), inhibitors or modulators of PDHK2 or PDHK4, inhibitors of MAP4K4, modulators of IL1 family including ILI beta, modulators of RXRalpha. In addition suitable anti-diabetic agents include mechanisms listed by Carpino, P. A., Goodwin, B. Expert Opin. Ther. Pat, 2010, 20(12), 1627-51 ;

an anti-platelet agent. Especially preferred anti-platelet agents are aspirin and clopidogrel. The term anti-platelet agents (or platelet inhibitory agents), as used herein, denotes agents that inhibit platelet function, for example by inhibiting the aggregation, adhesion or granular secretion of platelets. Agents include, but are not limited to, the various known non-steroidal anti-inflammatory drugs (NSAIDS) such as aspirin, ibuprofen, naproxen, sulindac, indomethacin, mefenamate, droxicam, diclofenac, sulfinpyrazone, piroxicam, and pharmaceutically acceptable salts or prodrugs thereof. Of the NSAIDS, aspirin (acetylsalicyclic acid or ASA) and COX-2 inhibitors such as celecoxib or piroxicam are preferred. Other suitable platelet inhibitory agents include llb/llla antagonists (e.g., tirofiban, eptifibatide, and abciximab), thromboxane-A2-receptor antagonists (e.g., ifetroban), thromboxane-A2-synthetase inhibitors, PDE3 inhibitors (e.g., Pletal, dipyridamole), and pharmaceutically acceptable salts or prodrugs thereof. The term anti-platelet agents (or platelet inhibitory agents), as used herein, is also intended to include ADP (adenosine diphosphate) receptor antagonists, preferably antagonists of the purinergic receptors P2Y1 and Ρ2Υ12· ^w'^tn ^2^12 being even more preferred. Preferred P2Y12 receptor antagonists include ticagrelor, prasugrel, ticlopidine and clopidogrel, including pharmaceutically acceptable salts or prodrugs thereof. Clopidogrel is an even more preferred agent. Ticlopidine and clopidogrel are also preferred compounds since they are known to be gentle on the gastro-intestinal tract in use;

an anti-thrombin agent including inhibitors of the serine protease thrombin. By inhibiting thrombin, various thrombin-mediated processes, such as thrombin-mediated platelet activation (that is, for example, the aggregation of platelets, and/or the granular secretion of plasminogen activator inhibitor-1 and/or serotonin) and/or fibrin formation are disrupted. A number of thrombin inhibitors are known to one of skill in the art and these inhibitors are contemplated to be used in combination with the present compounds. Such inhibitors include, but are not limited to, boroarginine derivatives, boropeptides, heparins, hirudin, argatroban, and melagatran, including pharmaceutically acceptable salts and prodrugs thereof. Boroarginine derivatives and boropeptides include N-acetyl and peptide derivatives of boronic acid, such as C- terminal alpha-aminoboronic acid derivatives of lysine, ornithine, arginine, homoarginine and corresponding isothiouronium analogs thereof. The term hirudin, as used herein, includes suitable derivatives or analogs of hirudin, referred to herein as hirulogs, such as disulfatohirudin;

a thrombolytics or fibrinolytic agents (or thrombolytics or fibrinolytics), including agents that lyse blood clots (thrombi). Such agents include tissue plasminogen activator (natural or recombinant) and modified forms thereof, anistreplase, urokinase, streptokinase, tenecteplase (TNK), lanoteplase (nPA), factor Vila inhibitors, PAI-1 inhibitors (i.e., inactivators of tissue plasminogen activator inhibitors), alpha2-antiplasmin inhibitors, and anisoylated plasminogen streptokinase activator complex, including pharmaceutically acceptable salts or prodrugs thereof. The term anistreplase, as used herein, refers to anisoylated plasminogen streptokinase activator complex, as described, for example, in EP 028,489, the disclosure of which is hereby incorporated herein by reference herein. The term urokinase, as used herein, is intended to denote both dual and single chain urokinase, the latter also being referred to herein as prourokinase. Examples of suitable anti-arrythmic agents include: Class I agents (such as propafenone); Class II agents (such as metoprolol, atenolol, carvadiol and propranolol); Class III agents (such as sotalol, dofetilide, amiodarone, azimilide and ibutilide); Class IV agents (such as ditiazem and verapamil); K⁺ channel openers such as \/ _c^ inhibitors, and IK_ui- inhibitors (e.g., compounds such as those disclosed in WO01/40231);

(viii) a factor Xa inhibitor, including apixaban and rivaroxaban;

(ix) an anti-coagulants for use in combination with the compounds of the present invention include heparins (e.g., unfractioned and low molecular weight heparins such as enoxaparin and dalteparin);

(x) other agents such as warfarin, unfractionated heparin, low molecular weight heparin, synthetic pentasaccharide, hirudin, argatrobanas, aspirin, ibuprofen, naproxen, sulindac, indomethacin, mefenamate, droxicam, diclofenac, sulfinpyrazone, piroxicam, ticlopidine, clopidogrel, tirofiban, eptifibatide, abciximab, melagatran, disulfatohirudin, tissue plasminogen activator, modified tissue plasminogen activator, anistreplase, urokinase, and streptokinase;

(xi) a neuroinflammatory or a neurodegenerative agent. Examples of neuroinflammatory and neurodegenerative agents include antidepressants, antipsychotics, anti-pain agents, anti-Alzheimer's agents, and anti-anxiety agents. Examples of particular classes of antidepressants that can be used in combination with the compounds useful for the present invention include norepinephrine reuptake inhibitors, selective serotonin reuptake inhibitors

(SSRIs), NK-1 receptor antagonists, monoamine oxidase inhibitors (MAOIs), reversible inhibitors of monoamine oxidase (RIMAs), serotonin and noradrenaline reuptake inhibitors (SNRIs), corticotropin releasing factor (CRF) antagonists, and atypical antidepressants. Suitable norepinephrine reuptake inhibitors include tertiary amine tricyclics and secondary amine tricyclics.

Examples of suitable tertiary amine tricyclics and secondary amine tricyclics include amitriptyline, clomipramine, doxepin, imipramine, trimipramine, dothiepin, butriptyline, nortriptyline, protriptyline, amoxapine, desipramine and maprotiline. Examples of suitable SSRIs include fluoxetine, fluvoxamine, paroxetine, and sertraline. Examples of monoamine oxidase inhibitors include isocarboxazid, phenelzine, and tranylcyclopramine. Examples of suitable reversible inhibitors of monoamine oxidase include moclobemide. Examples of suitable SNRIs of use in the present invention include venlafaxine. Examples of suitable atypical antidepressants include bupropion, lithium, trazodone and viloxazine. Examples of anti-Alzheimer's agents include NMDA receptor antagonists such as memantine; and cholinesterase inhibitors such as donepezil and galantamine. Examples of suitable classes of anti-anxiety agents that can be used in combination with the compounds useful for the present invention include benzodiazepines and serotonin 1A receptor (5-HT1A) agonists, and CRF antagonists. Suitable benzodiazepines include alprazolam, chlordiazepoxide, clonazepam, chlorazepate, diazepam, lorazepam, oxazepam, and prazepam. Suitable 5- HT1A receptor agonists include buspirone and ipsapirone. Suitable CRF antagonists include verucerfont. Suitable atypical antipsychotics include paliperidone, ziprasidone, risperidone, aripiprazole, olanzapine, and quetiapine. Suitable nicotine acetylcholine agonists include CP-601927 and varenicline.

Anti-pain agents include pregabalin, gabapentin, clonidine, neostigmine, baclofen, midazolam, ketamine and ziconotide;

(xii) an agent to reduce spasticity and spasms including, but not limited to, baclofen, tizanidine, benzodiazepines, dantrolene sodium, gabapentin and botulinum toxin; (xiii) an antioxidants, including but not limited to, myeloperoxidase inhibitors (such as AZD-3241), NOX4 and other NOX enzymes (such as GKT-137831), vitamin E, Coenzyme Q, and N-acetyl cysteine;

(xiv) a further BET-family bromodomain inhibitor;

(xv) a non-steroidal anti-inflammatory drugs (NSAIDs), including but not limited to, non-selective COX1/2 inhibitors such as piroxicam, naproxen, flubiprofen, fenoprofen, ketoprofen, ibuprofen, etodolac (Lodine), mefanamic acid, sulindac, apazone, pyrazolones (such as phenylbutazone), salicylates (such as aspirin); selective COX2 inhibitors such as: celecoxib, rofecoxib, etoricoxib, valdecoxib, meloxicam;

(xvi) an immunomodulatory and/ or anti-inflammatory agents, including but not limited to, methotrexate, leflunomide, ciclesonide chloroquine, hydroxychloroquine, d- penicillamine, auranofin, sulfasalazine, sodium aurothiomalate, cyclosporine, azathioprine, cromolyn, hydroxycarbamide, retinoids, fumarates (such as monomethyl and dimethyl fumarate), glatiramer acetate, mitoxantrone, teriflunomide, suplatast tosilate, mycophenolate mofetil and cyclophosphamide, laquinimod, voclosporin, PUR-118, AMG 357, AMG 81 1 , BCT197;

(xvii) a Nrf2 pathway activators, including but not limited to, fumarates, sulfurophane and bardoxolone methyl; and

(xviii) an interferon including, but not limited to, interferon beta-1 a and interferon beta- l b.

In one embodiment the additional therapeutic agent is an agent useful for treating Friedreich's ataxia; treating a condition or a disease associated with decreased expression of frataxin or a decreased level of frataxin; regulating the expression of frataxin in a cell; and / or for increasing the expression of frataxin in a cell. Such agents include, but are not limited to, those selected from the group consisting of carbanylated erythropoietin; interferon gammal b; methylprednisolone; idebenone; quinone antioxidants such as A00001 - alpha tocopherolquinone and EP1-743; iron chlating agents cuah as deferiprone; Nrf2 enhancers such as RTA-408; acetyl- 1 -carnitine; pioglitazone; di-deutero synthetic analogues of linoleic acid ethyl ester such as RT001 ; ginkgo biloba extract such as EGb761 ; free radical scavengers such as indole-3- propionic acid or EP20629; nicotinamide; and riluzole.

Inasmuch as it may be desirable to administer a combination of active compounds, for the pharmaceutical methods of the present invention, it is within the scope of the present invention that two or more pharmaceutical compositions may conveniently be combined in the form of a kit suitable for co-administration of the compositions.

BIOLOGICAL EXAMPLES

Test Compound Source

Test compounds JQ-1(+), JQ-1 (-), and l-BET-762 were obtained from Selleck Chemicals, Houston, Texas, USA. Other test compounds were obtained from Pfizer's library, which compounds had been prepared in accordance with methods from the art. For example RVX-208 was prepared in accordance with the method set out in WO2008/092231 ; l-BET-151 was prepared in accordance with the method set out in WO2011054843; N-(2-hydroxy-3-methylquinolin-6-yl)piperidine-1-sulfonamide was prepared in accordance with the method set out in WO2013/027168; N-[4-(2,4- difluorophenoxy)-3-(6-methyl-7-oxo-6,7-dihydro-1 H-pyrrolo[2,3-c]pyridin- 4- yl)phenyl]ethanesulfonamide was prepared in accordance with the method set out in WO 2013/097052 or WO 2013/097601 ; and N-{6-[methyl(propanoyl)amino]-3-oxo-4- [(1 S)-1-phenylpropyl]-3,4-dihydropyrido[2,3-b]pyrazin-2-yl}-beta-alanine was prepared in accordance with the method set out in US provisional patent application serial number 62/181281 , which was filed on 18^th June 2015.

All compounds were reconstituted in DMSO (Sigma) prior to use.

Example 1 - Bromodomain 4 (BRD4) - Fluorescence Polarization (FP) Binding Assay

The biological activity of BET-family bromodomain inhibitors useful for the present invention may be measured in the BRD4 Fluorescence polarization (FP) competition binding assay where the compounds can be screened for their activity at BRD4 BD1 , BRD4 BD2 or both BRD4 BD1 and BD2.

His-tagged BRD4 BD1 (44-160) and PFI-411 FP (Cy5-labeled FP probe) were prepared as previously described (Picaud S et al PFI-1 , a highly selective protein interaction inhibitor, targeting BET Bromodomains. Cancer Res. (2013) 73: 3336-46 and Wu, J et al Design and chemoproteomic functional characterization of a chemical probe targeted to bromodomains of BET-family proteins. Med. Chem. Commun. 2014, vol 5m p1871-1878, incorporated by reference herein). His-tagged BRD4 BD2 (342- 460) was purchased from BPS Bioscience. All assay components were diluted in 50 mM HEPES pH 7.4 containing 0.08% bovine serum albumin (assay buffer). To start the assay, 8 μΙ_ of the indicated BRD4 was added to each well of a low volume 384-well black flat bottom microtiter plate (Corning 3820) containing 4 μΙ of various concentrations of test compound (each plate also contained positive and negative control wells to define the upper and lower limits of the assay signal). After addition of BRD4, the assay plate was incubated at room temperature (RT, 20°C). After 15 minutes, 4 μΙ_ of PFI-411 FP was added to each well and the assay plate was placed in the dark at RT. The final assay concentration (FAC) of PFI-41 1 FP was 2 nM, the FAC of BRD4 BD1 and BRD4 BD2 was 50 nM, the FAC of test compound ranged from 120 to 0.0012 μΜ and the FAC of DMSO was 0.4%. After 60 min, polarization values were measured with an Envision 2103 multilabel reader (Perkin Elmer) using a Cy5 dual enhanced mirror and excitation at 620 nm and emission at 688 nm. The percent (%) effect was calculated for each concentration of test compound and was relative to the amount of polarization signal produced by the positive and negative control wells contained within each assay plate. The concentrations and % effect values for test compounds were plotted versus each other with a proprietary curve fitting program using a four-parameter logistic dose response equation and the concentration of compound required for 50% effect (IC50) was determined. The Ki values of competitive inhibitors are calculated using the equation described by Nikolovska-Coleska et al. (Development and optimization of a binding assay for the XIAP BIR3 domain using fluorescence polarization. Analytical Biochemistry (2004) 332: 261-273, incorporated by reference herein).

Certain compounds were tested in the Fluorescence Polarization assay described above:

Example 2 - Frataxin levels in Friedreich's Ataxia patient derived lymphoblasts are increased on treatment of with JQ-1 (+) and l-BET-762

Friedreich's Ataxia lymphoblasts GM 15850 (FXN alleles carrying ~ 650 and 1030 GAA repeats) were obtained from NIGMS Human Genetic Cell Repository at the Coriell Institute for Medical Research and cultured in complete medium containing RPMI1640 media supplemented with 10% Fetal bovine serum (Invitrogen) and 20mM Hepes buffer (Invitrogen). Prior to treatment with a test compound, 500,000 cells were seeded in 900ul complete medium were seeded per well in a 12 well plate, and subsequently treated with the test compound (10mM in DMSO) which had been diluted in 100ul optimem (Invitrogen) to achieve a final concentration of test compound of 5.0μΜ; 0.5μΜ or 0.05μΜ. Cells were incubated in a 37°C incubator (5% C0₂) for 72h before harvesting. For harvesting, cells were collected by centrifugation at 500xg for 5 mins, then washed once in 1 ml phosphate buffered saline (PBS) before lysis in NP40 lysis buffer (0.5%NP40+50mM Tris pH7.5, 150mM NaCI supplemented with protease and phosphatase inhibitors from Roche) for 30 mins on ice . Lysates were finally cleared by centrifugation at max speed in a refrigerated microcentrigue for 10 mins. Protein concentration was determined using the Protein 660 reagent (Pierce) (results shown in Figure 2, total protein concentration in mg/ml) and lysates were prepared for polyacrylamide gel electrophoresis (PAGE) analysis by dilution in lysis buffer supplemented with LDS sample buffer (Invitrogen) containing 10% dithiothreitol. Samples (5-10ug/lane) were then denatured by heating at 100C for 5 mins, loaded on a 4-12% BisTris Nupage gel (Invitrogen) and resolved in MES running buffer (Invitrogen) supplemented with Nupage antioxidant (Invitrogen). Proteins were transferred to a nitrocellulose membrane (0.2um from Biorad) using the Trans-Blot Turbo (Biorad). Membrane was blocked in 5% milk in Tris buffered saline supplemented with 0.05% Tween 20 (TBST) for 1 h and left in primary antibody anti-FXN (ab110328 from Abeam; 1 : 1000 dilution) diluted in blocking buffer overnight at 4C. Membrane was then washed three times in TBST and incubated in blocking buffer containing secondary anti-mouse antibody conjugated to HRP (1 :7500 anti-mouse IgG HRP) for 30 mins and then subjected to four washes in TBST. Similarly, anti^-actin (Cell Signaling; 1 :12000 dilution) western blotting was used as lane loading control. Signal was finally developed in ECL prime HRP substrate (GE) for 5 mins and membranes were exposed to Biomax film (Kodak) before processing in a film developer. Results (shown in Figure 1) from both a short and long exposure of the film are reported in Figure 1. Bands corresponding to both intermediate frataxin (i) and mature frataxin (m) are indicated in Figure 1.

The results of these experiments (Figure 1) demonstrate that treatment of GM 15850 cell line with BET-family bromodomain inhibitor JQ-1 (+) or IBET762 increased the levels of both intermediate and mature frataxin. Additionally treatment with the inactive enantiomer JQ-1 (+) had no significant effect on frataxin levels suggesting that the increase in frataxin is as a result of the inhibition of the BET-family bromodomain.

However, as shown in Figure 2, treatment of GM 15850 cell line with BET-family bromodomain inhibitor JQ-1 (+) or l-BET-762 induced cytotoxic effects as seen by reduction in the concentration of total cellular protein. This cytotoxic effect was further investigated, see Example 4 below.

Example 3 - The dose dependent effect of JQ-1(+) on frataxin levels in

Friedreich's Ataxia patient derived lymphoblasts was measured

GM 15850 cells (Coriell) were subjected to a dose-response treatment with JQ-

1 (+) in duplicate at concentrations that yield low cytotoxicity in duplicate, as described above in. The concentration of JQ-1(+) tested were 6.25nM, 12.5nM, 25nM and 50nM, chosen to be minimally cytotoxic level. As a reference for levels of frataxin in healthy cells, AG14725 lymphoblasts (Coriell) were treated, again in duplicate, with DMSO alongside. Lysates from both cell lines were subsequently analyzed by western blotting, as previously described in Example 2. Bands corresponding to both intermediate frataxin (i) and mature frataxin (m) are indicated in Figure 3. The results in Figure 3 show that treatment of GM 15850 cells with JQ-1 (+) at 25nM and 50nM concentration resulted in an increased in both intermediate and mature frataxin.

Bands corresponding to mature frataxin protein were quantified using Pixel Densitometry using the Image StudioLite (version 4.0) software from Licor. The results are shown in Figure 4. These results confirm that treatment of GM 15850 cells with JQ- 1 (+) at 25nM and 50nM concentration resulted in an increased in both intermediate and mature frataxin and the level of increase was quantified to be approximately 1.5 fold increase.

Example 4 - The cytotoxic effect of JQ-1(+) on healthy AG14725 lymphoblasts and Friedreich's Ataxia GM 15850 lymphoblasts was measured

This experiment was conducted to confirm that low levels of cellular frataxin was not sensitizing the test cell line to the cytotoxic effects induced by treatment with the BET-family bromodomain test compound, and that such cells were not being selectively killed to enrich for higher expressors. A dose response treatment of GM 15850 lymphoblasts (Coriell) and AG 14725 lymphoblasts (Coriell) was conducted with JQ-1 (+) to determine the Lethal Dose 50 (LD₅₀, concentration of JQ-1 (+) that yields 50% cell killing). The lymphoblasts were seeded in a 96 well plate, (40,000 cells per well in 180μΙ_ of RPMI1640 media supplemented with 10% Fetal bovine serum (Invitrogen) and 20mM Hepes buffer (Invitrogen)) and were incubated in a 37°C incubator (5% CO₂) for 72h before harvesting with varying concentrations of the test compound (duplicate treatment per condition). The test compound was prepared by dilution of a 10mM stock solution in DMSO with optimen (Invitrogen) to the desired test concentration. Finally, cell viability was determined using the CellTiter Glo luminescent assay (Promega) according to the protocol provided by the manufacturer. The results are shown in Figure 5. The results indicate that the LD50 displayed for both cell lines treated, namely the healthy AG14725 and FRDA GM15850, was approximately 100nM and was comparable for both cell lines. These results suggest that lymphoblasts containing reduced level of frataxin are not sensitized to killing by BET inhibitors.

Example 5 - Frataxin levels in Friedreich's Ataxia patient derived lymphoblasts are increased on treatment of with a variety of different BET-family bromodomain inhibitor scaffolds.

We further examined the effect on frataxin levels of a variety of small molecule compounds, known to have BET-family bromodomain inhibitory activity, and with a variety of chemical scaffolds. GM 15850 lymphoblasts were prepared according to the directions provided in Example 2 above and treated with a 1 μΜ solution of the test compound for 72 hours, as described previously. The test compound was prepared by dilution of a 10mM stock solution in DMSO with optimen (Invitrogen) to the desired test concentration. The compounds tested were:

The levels of intermediate and mature frataxin in the cells were assessed by Western Blotting using the same protocol as described in Example 2 above. Lysates were also subjected to Western Blotting again using the same protocol with an antibody raised against mitochondrial aconitase (mAco2); the antibody was obtained from Abeam (ab129105). The results (shown in Figure 6) demonstrate that the level of both intermediate and mature frataxin was increased for all compounds tested, although the increase seen with Cmpdl was reduced when compared to the other compounds tested which, without wishing to be bound by theory, may possibly be a result of the reduced solubility of Cmpdl in the test medium. Our data also shows that the levels of mAco2 remain unchanged by treatment with the test compound BET-family bromodomain inhibitors indicating that these compounds do not cause a global increase in levels of all mitochondrial proteins such as mAco2 and thus confirming that the effect seen was specific to frataxin. Example 6 - Bromodomain containing 4 protein (BRD4) knockdown and inhibition in Friedreich's Ataxia fibroblasts increases frataxin levels

Further experiments were conducted to validate that knockdown of the well characterized BET-family bromodomain protein BRD4 can reproduce the effect observed by chemical inhibition of BET, and to examine if this inhibition would be observed in other non-lymphoblast patient derived cells. GM03665 cells (Coriell; -700 and 400 GAA repeats) were seeded at 100,000 cells/well in a 6-well plate in 2ml_ of DMEM media (Invitrogen) supplemented with 10% Fetal bovine serum (Invitrogen) and 20mM Hepes buffer (Invitrogen)). The cells were reverse transfected using transfection reagent Dharmafect I (Dharmacon) following manufacturer's recommendation mixed with the following siRNA duplex at a concentration of 25nM in optimen (Invitrogen).

Additionally, one well was treated with 1 μΜ JQ-1 (+) prepared by dilution of a 10mM stock solution in DMSO with optimen (Invitrogen). All the cells were incubated for 72hrs following transfection or treatment, lysed and analysed by Western Blotting as previously described in Example 2 above. The results are shown in Figure 7. These results show that knockdown of BRD4 expression using the BRD4 siRNA smartpool resulted in an increase in both intermediate and mature frataxin levels. Furthermore the treatment with JQ-1 (+) (1 μΜ) resulted in an increase intermediate and mature frataxin levels demonstrating that the earlier results seen are not limited to the Friedreich's Ataxia lymphoblasts.

Example 7 - Knockdown of BET proteins upregulates FXN

Further experiments were conducted to validate that knockdown of all BET- family bromodomains can reproduce the effect observed by chemical inhibition of BET, and to examine if this inhibition would be observed in other non-lymphoblast patient derived cells. GM03665 cells (Coriell; -700 and 400 GAA repeats) were seeded at 100,000 cells/well in a 6-well plate in 2ml_ of DMEM media (Invitrogen) supplemented with 10% Fetal bovine serum (Invitrogen) and 20mM Hepes buffer (Invitrogen)). The cells were reverse transfected using transfection reagent Dharmafect I (Dharmacon) following manufacturer's recommendation mixed with the following siRNA duplex at a concentration of 25nM in optimen (Invitrogen).

All the cells were incubated for 72hrs following transfection or treatment, lysed and analysed by Western Blotting as previously described in Example 2 above. The results are shown in Figure 8. These results show that knockdown of all BRD subtypes expression using the siRNA smartpool resulted in an increase in both intermediate and mature frataxin levels.

Variations, modifications, and other implementations of what is described herein will occur to those skilled in the art without departing from the spirit and the essential characteristics of the present teachings. Accordingly, the scope of the present teachings is to be defined not by the preceding illustrative description but instead by the following claims, and all changes that come within the meaning and range of equivalency of the claims are intended to be embraced therein.

Each of the printed publications, including but not limited to patents, patent applications, books, technical papers, trade publications and journal articles described or referenced in this specification are herein incorporated by reference in their entirety and for all purposes.

Claims

1. A method of increasing the level of frataxin in a cell or a subject, comprising administering to the cell or the subject in need thereof a therapeutically effective amount of a BET- family bromodomain inhibitor, or a pharmaceutically acceptable salt thereof.

2. A method of increasing the expression of frataxin in a cell or a subject, comprising administering to the cell or the subject in need thereof a therapeutically effective amount of a BET- family bromodomain inhibitor, or a pharmaceutically acceptable salt thereof.

3. The method according to Claim 1 or Claim 2 wherein the BET- family bromodomain inhibitor is a BRD-4 inhibitor, or a pharmaceutically acceptable salt thereof.

4. The method according to Claim 3 wherein the BET- family bromodomain inhibitor is selected from the group consisting of l-BET-762;

JQ-1 ;

JQ-1 (+);

CPI-203;

OTX-015;

GW-841819X;

CP-0610;

CPI-232;

BET-BAY-002;

l-BET-151 ;

RVX-208;

l-BET-726;

SRX-2523; N-(2-hydroxy-3-methylquinolin-6-yl)piperidine-1 -sulfonamide;

N-[4-(2,4-difluorophenoxy)-3-(6-methyl-7-oxo-6,7-dihydro-1 H-pyrrolo[2,3-c]pyridin- 4-yl)phenyl]ethanesulfonamide; and

N-{6-[methyl(propanoyl)amino]-3-oxo-4-[(1 S)-1^henylpropyl]-3,4-dihydropyrido[2,3 b]pyrazin-2-yl}-beta-alanine

or a pharmaceutically acceptable salt thereof.

5. The method according to Claim 4, wherein the BET- family bromodomain inhibitor is l-BET-762, or a pharmaceutically acceptable salt thereof.

6. The method according to Claim 4, wherein the BET- family bromodomain inhibitor is JQ-1 (+), or a pharmaceutically acceptable salt thereof.

7. The method according to Claim 4, wherein the BET- family bromodomain inhibitor is l-BET-151 , or a pharmaceutically acceptable salt thereof.

8. The method according to Claim 4, wherein the BET- family bromodomain inhibitor is RVX-208, or a pharmaceutically acceptable salt thereof.

9. The method according to Claim 4, wherein the BET- family bromodomain inhibitor is N-(2-hydroxy-3-methylquinolin-6-yl)piperidine-1 -sulfonamide, or a pharmaceutically acceptable salt therof.

10. The method according to Claim 4, wherein the BET-family bromodomain inhibitor is N-[4-(2,4-difluorophenoxy)-3-(6-methyl-7-oxo-6,7-dihydro-1 H-pyrrolo[2,3-c]pyridin- 4- yl)phenyl]ethanesulfonamide, or a pharmaceutically acceptable salt thereof.

11. The method according to Claim 4, wherein the BET-family bromodomain inhibitor is N-{6-[methyl(propanoyl)amino]-3-oxo-4-[(1S)-1-phenylpropyl]-3,4-dihydropyrido[2,3 b]pyrazin-2-yl}-beta-alanine, or a pharmaceutically acceptable salt thereof.

12. A method of treating Friedreich's ataxia in a mammal comprising administering to a subject in need thereof a therapeutically effective amount of a BET-family bromodomain inhibitor, or a pharmaceutically acceptable salt thereof.

13. The method according to any one of Claims 12 wherein the BET- family bromodomain inhibitor is selected from the group consisting of l-BET-762;

JQ-1 ; JQ-1 (+);

CPI-203;

OTX-015;

GW-841819X;

CP-0610;

CPI-232;

BET-BAY-002;

l-BET-151 ;

RVX-208;

l-BET-726;

SRX-2523;

N-(2-hydroxy-3-methylquinolin-6-yl)piperidine-1 -sulfonamide;

N-{6-[methyl(propanoyl)amino]-3-oxo-4-[(1 S)-1-phenylpropyl]-3,4-dihydropyrido[2,3 b]pyrazin-2-yl}-beta-alanine

armaceutically acceptable salt thereof.