HK1242695B - Phthalazine derivatives of formula (i) as pcaf and gcn5 inhibitors for use in the treatment of cancer - Google Patents

Description

Phthaloazine derivatives of formula (I) as PCAF and GCN5 inhibitors for the treatment of cancer

CROSS-REFERENCE TO RELATED APPLICATIONS

This patent application claims priority to U.S. application No. 62/046,756, filed September 5, 2014, which is incorporated herein by reference.

Technical Field

The present invention relates to compounds useful as inhibitors of P300/CBP-associated factor (PCAF) and its closely related homolog GCN5, and methods of using such inhibitors to treat cancer.

Background Art

Chromatin is a complex combination of DNA and proteins that make up chromosomes. Found in the nucleus of eukaryotic cells, it is divided into heterochromatin (condensed) and euchromatin (extended) forms. The main components of chromatin are DNA and protein. Histones are the main protein components of chromatin and act as spools around which DNA is wound. The function of chromatin is to package DNA into a smaller volume to fit into the cell, strengthen DNA to allow mitosis and meiosis, and serve as a mechanism for controlling expression and DNA replication. Chromatin structure is controlled by a series of post-translational modifications of histone proteins (particularly histones H3 and H4), and the most common is the "histone tail" that extends beyond the core nucleosome structure. The histone tail is often subject to protein-protein interactions and is also the part of the histone most susceptible to post-translational modification. These modifications include acetylation, methylation, phosphorylation, ubiquitination, and sumoylation. These epigenetic marks are written and removed by specific enzymes that place tags on specific residues in the histone tails, thereby forming the epigenetic code, which is then interpreted by the cell to allow gene-specific regulation of chromatin structure and thus transcription.

Among all classes of proteins, histones are the most susceptible to post-translational modifications. Histone modifications are dynamic, because they can be added or removed in response to specific stimuli, and these modifications guide structural changes to chromatin and guide changes in gene transcription. Different classes of enzymes, i.e. histone acetyltransferases (HATs) and histone deacetylases (HDACs), acetylate or deacetylate specific histone lysine residues (Struhl K., Genes Dev., 1989, 12, 5, 599-606).

Bromodomains, approximately 110 amino acids long, are found in a large number of chromatin-associated proteins and have been identified in approximately 70 human proteins, often adjacent to other protein motifs (Jeanmougin F., et al., Trends Biochem. Sci., 1997, 22, 5, 151-153; and Tamkun J.W., et al., Cell, 1992, 7, 3, 561-572). The interaction between bromodomains and modified histones may be an important mechanism for causing changes in chromatin structure and gene regulation. Proteins containing bromodomains have been implicated in disease processes including cancer, inflammation, and viral replication. See, for example, Prinjha et al., Trends Pharm. Sci., 33(3): 146-153 (2012) and Muller et al., Expert Rev., 13(29): 1-20 (September 2011).

Cell-type specificity and proper tissue function require tight control of distinct transcriptional programs influenced by their environment. Alterations in this transcriptional homeostasis are directly linked to numerous disease states, most notably cancer, immune-inflammatory, neurological, and metabolic diseases. Bromodomains are located in key chromatin-modifying complexes that control unique disease-associated transcriptional pathways. Observations have highlighted that mutations in bromodomain-containing proteins are associated with cancer as well as immune and neurological dysfunction. Therefore, selective inhibition of bromodomains across specific families, such as those of PCAF, creates a wide range of opportunities as novel therapeutic agents for human disorders.

There is a need for treatments for cancer, immune disorders, and other PCAF bromodomain-associated diseases. There is also a need for treatments for GCN5-mediated disorders.

Summary of the Invention

One aspect includes compounds of formula (I) or salts thereof:

in:

R ¹ is selected from H, C _1-6 alkyl, C _2-6 alkenyl, C _2-6 alkynyl, carbocyclyl and heterocyclyl, wherein each C _1-6 alkyl, C _2-6 alkenyl, C _2-6 alkynyl, carbocyclyl and heterocyclyl is optionally substituted with one or more groups independently selected from the group consisting of oxo, halogen, amino, hydroxyl and C _1-6 alkoxy;

X is O, N(R ^a ) or S;

each ^Ra is independently selected from the group consisting of hydrogen, _C1-6 alkyl, _C2-6 alkenyl, _C2-6 alkynyl, _C1-6 alkoxy, and carbocyclyl, wherein each _C1-6 alkyl, _C2-6 alkenyl, _C2-6 alkynyl, C1-6 alkoxy, and carbocyclyl is optionally substituted with one or more groups independently selected from the group consisting of oxo, halogen, amino, hydroxy, _C1-6 alkoxy, and C1 _{-C6 alkyl} optionally substituted with one or more groups independently selected from the group consisting of oxo _and halogen;

R ³ and R ⁴ are each independently selected from hydrogen, C _1-6 alkyl, C _2-6 alkenyl, C _2-6 alkynyl, C _1-6 alkoxy, carbocyclyl and heterocyclyl, wherein each C _1-6 alkyl, C _2-6 alkenyl, C _2-6 alkynyl, C _1-6 alkoxy, carbocyclyl and heterocyclyl is optionally substituted with one or more groups R ^b ; or R ³ and R ⁴ together with the atoms to which they are attached form a carbocyclyl or heterocyclyl, said carbocyclyl or heterocyclyl being optionally substituted with one or more groups R ^b ;

R ⁵ is hydrogen or C _1-6 alkyl, or R ⁴ and R ⁵ together with the atoms to which they are attached form a carbocyclyl or heterocyclyl, said carbocyclyl or heterocyclyl being optionally substituted with one or more groups independently selected from the group consisting of C _1-6 alkyl, C _2-6 alkenyl, C _2-6 alkynyl, C _1-6 alkoxy, C _3-6 carbocyclyl, oxo, halogen, hydroxy and -NO ₂ , wherein any of the C _1-6 alkyl, C _2-6 alkenyl, C _2-6 alkynyl, C _1-6 alkoxy and C _3-6 carbocyclyl groups are optionally substituted with one or more groups independently selected from the group consisting of halogen, hydroxy and C _1-6 alkoxy;

Each ^Rb is independently selected from _C1-6 alkyl, _C2-6 alkenyl, _C2-6 alkynyl, carbocyclyl, heterocyclyl, oxo, halogen, -NO2, -N( ^Rc ) ₂ , -CN, -C(O)-N( ^Rc ) ₂ , -S(O)-N( ^Rc ) ₂ , -S(O) ₂ -N( ^Rc ) _{2 ,} ^-ORc , ^-SRc , -OC(O) ^-Rc , -C(O)-Rc, -C(O) ^-ORc , ^-S (O) ^-Rc , -S(O) ₂ - ^Rc , -C(O)-N( ^Rc ) ₂ , -N( ^Rc )-C(O) ^-Rc , -N( ^Rc )-S(O) ^-Rc , and -N( ^Rc )-S(O) ₂ - ^Rc , wherein any C _C1-6 alkyl, _C2-6 alkenyl, _C2-6 alkynyl, carbocyclyl and heterocyclyl are optionally substituted with one or more groups independently selected from the group consisting of oxo, halogen, -NO2, -N( ^Rc ) ₂ , -CN, -C(O)-N( ^Rc ) ₂ , -S(O)-N( ^Rc ) ₂ , -S(O) _{2 -} N( ^Rc ) ₂ , ^-ORc , ^-SRc , -OC(O) ^-Rc , -C(O) ^-Rc , -C(O) ^-ORc , -S(O) ^-Rc , -S(O) _2- ^Rc , -C(O)-N( ^Rc ) ₂ , -N( ^Rc )-C(O) ^-Rc , -N( ^Rc )-S(O) ^-Rc and -N( ^Rc )-S(O) ₂ - ^Rc ;

each R ^c is independently selected from the following groups: hydrogen, C _1-6 alkyl, C _2-6 alkenyl, C _2-6 alkynyl, C _1-6 alkoxy, carbocyclyl and heterocyclyl, wherein each C _1-6 alkyl, C _{2-6 alkenyl, C 2-6 alkynyl, C 1-6 alkoxy} , carbocyclyl and heterocyclyl is optionally substituted with one or more groups independently selected from the following groups: oxo, halogen, amino, hydroxyl, C _1-6 alkoxy, carbocyclyl, heterocyclyl and C ₁ -C ₆ alkyl optionally substituted with one or more groups independently selected from oxo and halogen; or two R ^c together with the nitrogen to which they are attached form heterocyclyl optionally substituted with one or more groups independently selected from oxo and halogen and C _1-3 alkyl optionally substituted with one or more groups independently selected from oxo and halogen;

Ring A is a 5-membered or 6-membered heterocyclyl or a 5-membered or 6-membered carbocyclyl, wherein the 5-membered or 6-membered heterocyclyl and the 5-membered or 6-membered carbocyclyl are optionally substituted with one or more groups independently selected from the group consisting of C _1-6 alkyl, C _2-6 alkenyl, C _2-6 alkynyl, carbocyclyl, heterocyclyl, halogen, -NO ₂ , -N(R ^d ) ₂ , -CN, -C(O)-N(R ^d ) ₂ , -S(O)-N(R ^d ) ₂ , -S(O) ₂ -N(R ^d ) ₂ , -OR ^d , -SR ^d , -OC(O)-R ^d , -C(O)-R ^d , -C(O)-OR ^d , -S(O)-R ^d , -S(O) ₂ -R ^d , -C(O)-N(R ^d ) ₂ d ) , -N(R ^d )-C(O)-R ^d , -N(R ^d )-S(O)-R ^d , -OC(O)-N(R ^d ) ₂ , -N(R ^d )-C(O)-OR ^d , -N(R ^d )-C(O)-N(R ^d ) ₂ and -N(R ^d )-S(O) ₂ -R ^d , wherein any C _1-6 alkyl, C _2-6 alkenyl, C _2-6 alkynyl, carbocyclyl and heterocyclyl are optionally substituted with one or more groups independently selected from the group consisting of oxo, halogen, -NO ₂ , -N(R d ) 2 , -CN, -C(O)-N(R ^d ) 2 , -S(O)-N(R d ) ₂ , -S(O) 2 -N(R ^d ) ₂ , -OR d , -N(R d )-C(O)-N(R ^d ) ₂ and -N(R d ) _-S ( ^O ) ₂ -R ^{d .} , -SR ^d , -OC(O)-R ^d , -C(O)-R ^d , -C(O)-OR ^d , -S(O)-R ^d , -S(O) ₂ -R ^d , -C(O)-N(R ^d ) ₂ , -N(R ^d )-C(O)-R ^d , -N(R ^d )-S(O)-R ^d , -OC(O)-N(R ^d ) ₂ , -N(R ^d )-C(O)-OR ^d , -N(R ^d )-C(O)-N(R ^d ) ₂ and -N(R ^d )-S(O) ₂ -R ^d ;

Each R ^d is independently selected from the following groups: hydrogen, C _1-6 alkyl, C _2-6 alkenyl, C _2-6 alkynyl, C _1-6 alkoxy, carbocyclyl and heterocyclyl, wherein each C _1-6 alkyl, C _2-6 alkenyl, C _{2-6 alkynyl, C 1-6 alkoxy} , carbocyclyl and heterocyclyl is optionally substituted with one or more groups R ^f ; or two R ^d together with the nitrogen to which they are attached form a heterocyclyl optionally substituted with one or more groups R ^f ;

each R ^e is independently selected from the group consisting of hydrogen, C _1-6 alkyl, C _2-6 alkenyl, C _2-6 alkynyl, C _1-6 alkoxy, carbocyclyl, and heterocyclyl, wherein each C _1-6 alkyl, C _2-6 alkenyl, C _2-6 alkynyl, C _1-6 alkoxy, carbocyclyl, and heterocyclyl is optionally substituted with one or more groups independently selected from the group consisting of oxo, halogen, amino, hydroxy, C _1-6 alkoxy, carbocyclyl, heterocyclyl, and C ₁ -C ₆ alkyl optionally substituted with one or more groups independently selected from the group consisting of oxo and halogen;

or two ^Re together with the nitrogen to which they are attached form a heterocyclyl optionally substituted with one or more groups independently selected from oxo and halogen, and a _C1-3 alkyl optionally substituted with one or more groups independently selected from oxo and halogen; or one ^Re together with ^R3 and the atom to which they are attached form a heterocyclyl; optionally substituted with one or more groups independently selected from the following: _C1-6 alkyl, _C2-6 alkenyl, _C2-6 alkynyl and _C1-6 alkoxy, wherein each _C1-6 alkyl, _C2-6 alkenyl, _C2-6 alkynyl and _C1-6 alkoxy is optionally substituted with one or more groups independently selected from the following: oxo, halogen, amino, hydroxyl, _C1-6 alkoxy and carbocyclyl;

or one R ^e and R ⁴ together with the atoms to which they are attached form a heterocyclyl; optionally substituted with one or more groups independently selected from the following: C _1-6 alkyl, C _2-6 alkenyl, C _2-6 alkynyl and C _1-6 alkoxy, wherein each C _1-6 alkyl, C _2-6 alkenyl, C _2-6 alkynyl and C _1-6 alkoxy is optionally substituted with one or more groups independently selected from oxo, halogen, amino, hydroxy, C _1-6 alkoxy and carbocyclyl;

Each ^Rf is independently selected from _C1-6 alkyl, _C2-6 alkenyl, _C2-6 alkynyl, carbocyclyl, heterocyclyl, oxo, halogen, -NO2, -N( ^Rg ) ₂ , -CN, -C(O)-N( ^Rg ) ₂ , -S(O)-N(Rg) ₂ , -S(O) ₂ -N( ^Rg ) _{2 ,} ^-ORg , ^-SRg , -OC ⁽ O) ^-Rg , -C(O) ^-Rg , -C(O) ^-ORg , -S(O) ^-Rg , -S(O) ₂ - ^Rg , -N( ^Rg )-C(O) ^-Rg , -N( ^Rg )-S(O) ^-Rg , -N( ^Rg )-C(O)-N( ^Rg ) ₂ and -N( ^Rg )-S(O) ₂ -R ^g , wherein any _C1-6 alkyl, _C2-6 alkenyl, _C2-6 alkynyl, carbocyclyl and heterocyclyl are optionally substituted with one or more groups independently selected from the group consisting of oxo, halogen, -NO2, -N( ^Rg ) ₂ , -CN, -C(O)-N( ^Rg ) ₂ , -S(O)-N( ^Rg ) ₂ , -S(O) _{2 -} N( ^Rg ) ₂ , ^-ORg , ^-SRg , -OC(O)-Rg, -C(O) ^-Rg , -C(O) ^-ORg , -S(O) ^-Rg , -S(O) ₂ ^{-Rg ,} -N( ^Rg )-C(O) ^-Rg , -N( ^Rg )-S(O) ^-Rg , -N( ^Rg )-C(O)-N( ^Rg ) ₂ and -N( ^Rg )-S(O) ₂ -R ^g ; and

Each ^Rg is independently selected from the following groups: hydrogen, _C1-6 alkyl, _C2-6 alkenyl, _C2-6 alkynyl, _C1-6 alkoxy, carbocyclyl and heterocyclyl, wherein each _C1-6 alkyl, C2-6 alkenyl, _C2-6 alkynyl, _C1-6 alkoxy, carbocyclyl and heterocyclyl is optionally substituted with one or more groups independently selected from the following groups: oxo, halogen, amino, hydroxyl, _C1-6 alkoxy, carbocyclyl, heterocyclyl and C1- _C6 alkyl optionally substituted with one or more groups independently selected from oxo and halogen; or two ^Rg are taken together with the nitrogen to which they are attached to form heterocyclyl optionally substituted with one or more groups independently selected _from oxo, halogen and _{C1-3 alkyl} optionally substituted with one or more groups independently selected from oxo and halogen.

Another aspect includes compositions comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable adjuvant, carrier, or vehicle.

Another aspect includes a method for treating a PCAF-mediated disorder or a GCN5-mediated disorder in an animal (eg, a mammal such as a human) comprising administering to the animal a compound of Formula (I) or a pharmaceutically acceptable salt thereof.

Another aspect includes a compound of formula (I), or a pharmaceutically acceptable salt thereof, for use in medical therapy.

Another aspect includes a compound of formula (I), or a pharmaceutically acceptable salt thereof, for use in the prophylactic or therapeutic treatment of a PCAF-mediated disorder.

Another aspect includes the use of a compound of formula (I) or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating a PCAF-mediated disorder in an animal (eg, a mammal such as a human).

Another aspect includes compounds useful in PCAF studies.

Another aspect includes novel compounds of formula (I) and salts thereof.

Another aspect includes methods of a) increasing the efficacy of a cancer therapy comprising a cytotoxic agent in an animal (e.g., a mammal such as a human), b) delaying or preventing the development of cancer resistance to a cytotoxic agent in an animal (e.g., a mammal such as a human), or c) prolonging the duration of response to a cancer therapy in an animal (e.g., a mammal such as a human), comprising administering to the animal an effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof.

Another aspect includes a compound of formula (I) as described in claim 1, or a pharmaceutically acceptable salt thereof, for use in a) increasing the efficacy of a cancer treatment comprising a cytotoxic agent in an animal (e.g., a mammal such as a human), b) delaying or preventing the development of cancer resistance to a cytotoxic agent in an animal (e.g., a mammal such as a human), or c) prolonging the duration of response to a cancer therapy in an animal (e.g., a mammal such as a human).

Another aspect includes the use of a compound of formula (I), or a pharmaceutically acceptable salt thereof, in the preparation of a medicament for a) increasing the efficacy of a cancer treatment comprising a cytotoxic agent in an animal (e.g., a mammal such as a human), b) delaying or preventing the development of cancer resistance to a cytotoxic agent in an animal (e.g., a mammal such as a human), or c) prolonging the duration of response to a cancer therapy in an animal (e.g., a mammal such as a human).

Another aspect includes methods of treating cancer in an individual (eg, a patient) comprising administering to the individual (a) a compound of Formula (I) or a pharmaceutically acceptable salt thereof and (b) a cytotoxic agent.

Another aspect includes a compound of formula (I), or a pharmaceutically acceptable salt thereof, in combination with a cytotoxic agent for use in the prophylactic or therapeutic treatment of cancer.

Another aspect includes the use of a compound of formula (I) or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for use in combination with a cytotoxic agent in the treatment of cancer.

Another aspect includes a method for treating a GCN5-mediated disorder in an animal (eg, a mammal such as a human) comprising administering to the animal a compound of Formula (I) or a pharmaceutically acceptable salt thereof.

Another aspect includes a compound of formula (I), or a pharmaceutically acceptable salt thereof, for use in the prophylactic or therapeutic treatment of a GCN5-mediated disorder.

Another aspect includes the use of a compound of formula (I) or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating a GCN5-mediated disorder in an animal (eg, a mammal such as a human).

Another aspect includes compounds useful in the study of GCN5.

Another aspect includes synthetic intermediates and synthetic methods disclosed herein that are useful for preparing compounds of formula (I) or salts thereof.

DETAILED DESCRIPTION

Compounds and Definitions

Definitions and terminology are described in more detail below. Chemical elements are identified according to the Periodic Table of the Elements, CAS version, Handbook of Chemistry and Physics, 75th edition.

Unless otherwise indicated, compounds of Formula I include enantiomers, diastereomers, and geometric isomers (or conformations) of a given structure. For example, R and S configurations, Z and E double bond isomers, and Z and E conformations for each asymmetric center are included, as well as enantiomers, diastereomers, and geometric isomers (or conformations) mixtures. Unless otherwise indicated, all tautomeric forms of the structures described herein are included. Additionally, unless otherwise indicated, the structures described herein are also intended to include compounds differing only in the presence of one or more isotopically enriched atoms. For example, compounds of Formula I are included, wherein one or more hydrogens are replaced by deuterium or tritium, carbon by ¹³ C- or ¹⁴ C carbon, nitrogen by ¹⁵ N nitrogen, sulfur by ³³ S, ³⁴ S or ³⁶ S sulfur, oxygen by ¹⁷ O or ¹⁸ O oxygen, or fluorine by ¹⁸ F fluorine, independently replaced or enriched. Such compounds can be used as, for example, analytical tools, probes, or therapeutic agents in biological assays.

Where a specific enantiomer is described, in certain embodiments a material that is substantially free of the corresponding enantiomer may be provided and may also be referred to as "optically enriched". As used herein, "optically enriched" means that the mixture of enantiomers is composed of a significantly greater proportion of one enantiomer and may be described by enantiomeric excess (ee%). In certain embodiments, the mixture of enantiomers is composed of at least about 90% by weight of a given enantiomer (about 90% ee). In other embodiments, the mixture of enantiomers is composed of at least about 95%, 98% or 99% by weight of a given enantiomer (about 95%, 98% or 99% ee). Enantiomers and diastereomers may be separated from the racemic mixture by any method known to those skilled in the art, including recrystallization from a solvent in which one stereoisomer is more soluble than the other stereoisomer, chiral high pressure liquid chromatography (HPLC), supercritical fluid chromatography (SFC), formation and crystallization of chiral salts, followed by separation by any of the above methods, or prepared by asymmetric synthesis and optionally further enriched. See, for example, Jacques et al., Enantiomers, Racemates and Resolutions (Wiley Interscience, New York, 1981); Wilen, et al., Tetrahedron 33:2725 (1977); Eliel, E.L. Stereochemistry of Carbon Compounds (McGraw-Hill, NY, 1962); Wilen, S.H. Tables of Resolving Agents and Optical Resolutions p.268 (E.L. Eliel, Ed., Univ. of Notre Dame Press, Notre Dame, IN 1972).

The term "heteroatom" means any atom independently selected from other than carbon or hydrogen, such as one or more of oxygen, sulfur, nitrogen, phosphorus or silicon (including any oxidized form of nitrogen, sulfur, phosphorus or silicon; and any quaternized form of nitrogen).

As used herein, the terms "halogen" and "halogen" refer to an atom selected from fluorine (fluoro, -F), chlorine (chloro, -Cl), bromine (bromo, -Br) and iodine (iodo, -I).

The term "oxo" refers to =0.

As used herein, the term "unsaturated" means that the moiety has one or more units of unsaturation.

The term "carbocyclyl," used alone or as part of a larger moiety, refers to a saturated, partially unsaturated, or aromatic (e.g., aryl) ring system having 3 to 20 carbon atoms. In one embodiment, the carbocyclyl comprises 3 to 12 carbon atoms ( _C3 - _C12 ). In another embodiment, the carbocyclyl comprises _C3 - _C8 , _C3 - _C10 , or _C5 - _C10 . In other embodiments, the carbocyclyl as a monocyclic ring comprises _C3 - _C8 , _C3 - _C6 , or _C5 - _C6 . In another embodiment, the carbocyclyl as a bicyclic ring comprises C7- _C12 . In another embodiment, _the carbocyclyl as a spirocyclic ring system comprises _C5 - _C12 . Examples of monocyclic carbocyclic groups include cyclopropyl, cyclobutyl, cyclopentyl, 1-cyclopent-1-enyl, 1-cyclopent-2-enyl, 1-cyclopent-3-enyl, cyclohexyl, perdeuterated cyclohexyl, 1-cyclohex-1-enyl, 1-cyclohex-2-enyl, 1-cyclohex-3-enyl, cyclohexadienyl, cycloheptyl, cyclooctyl, cyclononyl, cyclodecyl, cycloundecyl, phenyl, and cyclododecyl; groups having 7 to 12 rings. Bicyclic carbocyclyls of atoms include [4,3], [4,4], [4,5], [5,5], [5,6] or [6,6] ring systems, for example bicyclo[2.2.1]heptane, bicyclo[2.2.2]octane, naphthalene and bicyclo[3.2.2]nonane; and spirocarbocyclyls spiro[2.2]pentane, spiro[2.3]hexane, spiro[2.4]heptane, spiro[2.5]octane and spiro[4.5]decane. The term carbocyclyl includes aryl ring systems as defined herein. The term carbocycle also includes cycloalkyl rings (e.g., saturated or partially unsaturated mono-, bi- or spiro-carbocycles).

As used herein, the term "alkyl" refers to a saturated straight or branched hydrocarbon group. In one embodiment, the alkyl group is one to eighteen carbon atoms ( _C1 - _C18 ). In other embodiments, the alkyl group is _C0 - _C6 , _C0 - _C5 , _C0 - _C3 , _C1 - _C12 , _C1 - _C10 , _C1 - _C8 , _C1 - _C6 , _C1 - _C5 , _C1 - _C4 , or _C1 - _C3 . _C0 alkyl refers to a bond. Examples of alkyl groups include methyl (Me, -CH ₃ ), ethyl (Et, -CH ₂ CH ₃ ), 1-propyl (n-Pr, n-propyl, -CH ₂ CH ₂ CH ₃ ), 2-propyl (i-Pr, i-propyl, -CH(CH ₃ ) ₂ ), 1-butyl (n-Bu, n-butyl, -CH ₂ CH ₂ CH ₂ CH ₃ ), 2-methyl-1-propyl (i-Bu, i-butyl, -CH ₂ CH(CH ₃ ) ₂ ), 2-butyl (s-Bu, s-butyl, -CH(CH ₃ )CH ₂ CH ₃ ), 2-methyl-2-propyl (t-Bu, t-butyl, -C(CH ₃ ) ₃ ), 1-pentyl (n-pentyl, -CH ₂ CH ₂ CH ₂ CH ₂ CH ₃ ), 2-pentyl (-CH(CH _{3 ) 2} )CH ₂ CH ₂ CH ₃ ), 3-pentyl (-CH(CH ₂ CH ₃ ) ₂ ), 2-methyl-2-butyl (-C(CH ₃ ) ₂ CH ₂ CH ₃ ), 3-methyl-2-butyl (-CH(CH ₃ )CH(CH ₃ ) ₂ ), 3-methyl-1-butyl (-CH ₂ CH ₂ CH(CH ₃ ) ₂ ), 2-methyl-1-butyl (-CH ₂ CH(CH ₃ )CH ₂ CH ₃ ), 1-hexyl (-CH ₂ CH ₂ CH ₂ CH ₂ CH ₃ ) _, 2-hexyl (-CH(CH ₃ )CH ₂ CH ₂ CH ₂ CH ₃ ), 3-hexyl (-CH(CH ₂ CH ₃ )(CH ₂ CH ₂ CH ₃ )), 2-methyl-2-pentyl (-C(CH ₃ ) ₂ CH _{2 3 ) C ( CH 3 ) 3 , heptyl , octyl , nonyl , decyl , undecyl and dodecyl . ​ ​ ​ ​}

As used herein, the term "alkenyl" refers to a straight or branched hydrocarbon group having at least one carbon-carbon double bond. Alkenyl groups include groups having "cis" and "trans" orientations, or "E" and "Z" orientations. In one example, an alkenyl group has 2 to 18 carbon atoms ( _C2 - _C18 ). In other examples, an alkenyl group is _C2 - _C12 , _C2 - _C10 , _C2 - _C8 , _C2 - _C6 , or _C2 - _C3 . Examples include, but are not limited to, ethenyl or vinyl (—CH═CH ₂ ), prop-1-enyl (—CH═CHCH ₃ ), prop-2-enyl (—CH ₂ CH═CH ₂ ), 2-methylprop-1-enyl, but-1-enyl, but-2-enyl, but-3-enyl, buta-1,3-dienyl, 2-methylbuta-1,3-dienyl, hex-1-enyl, hex-2-enyl, hex-3-enyl, hex-4-enyl, and hexa-1,3-dienyl.

As used herein, the term "alkynyl" refers to a straight or branched hydrocarbon group having at least one carbon-carbon triple bond. In one example, the alkynyl group is 2 to 18 carbon atoms ( _C2 - _C18 ). In other examples, the alkynyl group is _C2 - _C12 , _C2 - _C10 , _C2 - _C8 , _C2 - _C6 , or _C2 - _C3 . Examples include, but are not limited to, ethynyl (-C≡CH), prop-1-ynyl (-C≡CCH3), prop _- 2-ynyl (propargyl, _-CH2C≡CH ), but-1-ynyl, but-2-ynyl, and but-3-ynyl.

The term "alkoxy" refers to a straight or branched chain group represented by the formula -OR, wherein R is an alkyl, alkenyl, alkynyl or carbocyclic group. Alkoxy includes methoxy, ethoxy, propoxy, isopropoxy and cyclopropyloxy.

As used herein, the term "haloalkyl" refers to an alkyl group, as defined herein, substituted with one or more (eg, 1, 2, 3, or 4) halo groups.

The term "aryl," used alone or as part of a larger moiety as in "arylalkyl,""arylalkoxy," or "aryloxyalkyl," refers to a monocyclic, bicyclic, or tricyclic carbocyclic ring system, including fused rings, in which at least one ring in the system is aromatic. The term "aryl" is used interchangeably with the term "aryl ring." In one embodiment, aryl includes groups having 6-20 carbon atoms ( _C6 - _C20 aryl). In another embodiment, aryl includes groups having 6-10 carbon atoms ( _C6 - _C10 aryl). Examples of aryl groups include phenyl, naphthyl, anthracenyl, biphenyl, phenanthrenyl, naphthacene, 1,2,3,4-tetrahydronaphthyl, 1H-indenyl, 2,3-dihydro-1H-indenyl, and the like, which may be substituted or independently substituted with one or more substituents described herein. A specific aryl group is phenyl. In another embodiment, aryl includes an aryl ring fused to one or more carbocyclic rings, such as indenyl, phthalimidyl, naphthimidyl, phenantriidinyl, or tetrahydronaphthyl, and the like, wherein the radical or point of attachment is on the aromatic ring.

The term "heteroaryl," used alone or as part of a larger moiety, such as "heteroarylalkyl" or "heteroarylalkoxy," refers to a monocyclic, bicyclic, or tricyclic ring system having from 5 to 14 ring atoms, wherein at least one ring is aromatic and contains at least one heteroatom. In one embodiment, heteroaryl includes 4-6 membered monocyclic aromatic groups, wherein one or more ring atoms are independently optionally substituted nitrogen, sulfur, or oxygen. In another embodiment, heteroaryl includes 5-6 membered monocyclic aromatic groups, wherein one or more ring atoms are independently optionally substituted nitrogen, sulfur, or oxygen. In some embodiments, heteroaryl is C ₁ -C ₂₀ heteroaryl, wherein the heteroaryl ring contains 1-20 carbon atoms and the remaining ring atoms include one or more nitrogen, sulfur, or oxygen atoms. Examples of heteroaryl groups include thienyl, furyl, imidazolyl, pyrazolyl, thiazolyl, isothiazolyl, oxazolyl, isoxazolyl, triazolyl, thiadiazolyl, oxadiazolyl, tetrazolyl, thiatriazolyl, oxatriazolyl, pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, triazinyl, tetrazolyl, tetrazolyl [1,5-b] pyridazinyl, imidazo [1,2-a] pyrimidinyl, purinyl, benzoxazolyl, benzofuranyl, benzothiazolyl, benzo Thiadiazolyl, benzotriazolyl, benzimidazolyl, indolyl, 1,3-thiazol-2-yl, 1,3,4-triazol-5-yl, 1,3-oxazol-2-yl, 1,3,4-oxadiazol-5-yl, 1,2,4-oxadiazol-5-yl, 1,3,4-thiadiazol-5-yl, 1H-tetrazol-5-yl, 1,2,3-triazol-5-yl, pyridin-2-yl N-oxide and pyrazolo[4,3-c]pyridinyl. The term "heteroaryl" also includes groups in which the heteroaryl is fused to one or more aryl, carbocyclyl or heterocyclyl rings, wherein the radical or point of attachment is on the heteroaryl ring. Non-limiting examples include indolyl, isoindolyl, benzothiophenyl, benzofuranyl, dibenzofuranyl, indazolyl, benzimidazolyl, benzothiazolyl, quinolyl, isoquinolyl, cinnolinyl, phthalazinyl, quinazolinyl, quinoxalinyl, 4H-quinolizinyl, carbazolyl, acridinyl, phenazinyl, phenothiazinyl, phenoxazinyl, tetrahydroquinolyl and tetrahydroisoquinolyl. The heteroaryl group can be monocyclic, bicyclic or tricyclic.

The term "heterocyclyl" as used herein refers to a "carbocyclyl" as defined herein, wherein one or more (e.g., 1, 2, 3, or 4) carbon atoms have been replaced by heteroatoms (e.g., O, N, or S). In some embodiments, heterocyclyl refers to a saturated ring system, such as a 3- to 12-membered saturated heterocyclyl ring system. In some embodiments, heterocyclyl refers to a heteroaryl ring system, such as a 5-14-membered heteroaryl ring system. Heterocyclyl may be optionally substituted with one or more substituents independently selected from those defined herein.

In one example, the heterocyclic radical includes 3-12 ring atoms and includes monocycles, bicycles, tricycles and spirocycle systems, wherein the ring atoms are carbon, and one to five ring atoms are heteroatoms selected from nitrogen, sulfur or oxygen, which are independently optionally substituted by one or more groups. In one example, the heterocyclic radical includes 1 to 4 heteroatoms. In another example, the heterocyclic radical includes 3 to 7 membered monocycles with one or more heteroatoms selected from nitrogen, sulfur or oxygen. In another example, the heterocyclic radical includes 4 to 6 membered monocycles with one or more heteroatoms selected from nitrogen, sulfur or oxygen. In another example, the heterocyclic radical includes 3 membered monocycles. In another example, the heterocyclic radical includes 4 membered monocycles. In another example, the heterocyclic radical includes 5-6 membered monocycles. In one example, the heterocyclic radical includes 0 to 3 double bonds. Any nitrogen or sulfur heteroatom may optionally be oxidized (eg, NO, SO, SO ₂ ), and any nitrogen heteroatom may optionally be quaternized (eg, [NR ₄ ] ⁺ Cl ⁻ , [NR ₄ ] ⁺ OH ⁻ ). Exemplary heterocyclic groups include oxiranyl, aziridine, thiirane, azetidinyl, oxetanyl, thietanyl, 1,2-dithietanyl, 1,3-dithietanyl, pyrrolidinyl, dihydro-1H-pyrrolyl, dihydrofuranyl, tetrahydrofuranyl, dihydrothiophenyl, tetrahydrothiophenyl, imidazolidinyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, 1,1-dioxo-thiomorpholinyl, dihydropyranyl, tetrahydropyranyl, hexahydrothiopyranyl, hexahydropyrimidinyl, oxazinanyl, thiazinanyl, thioxanyl, homopiperazinyl, homopiperidinyl, pyridinyl, azepanyl, oxepanyl, thiepanyl, oxazepinyl, oxazepanyl, diazepanyl, 1,4-diazepanyl, diazepinyl, thiazepinyl, thiazepanyl, tetrahydrothiopyranyl, oxazolidinyl, thiazolidinyl, isothiazolidinyl, 1,1-dioxoisothiazolidinone, oxazolidinone, imidazolidinone, 4,5,6,7-tetrahydro[2H]indazolyl, tetrahydrobenzimidazolyl, 4,5,6,7-tetrahydrobenzo[d]imidazolyl, 1,6-diimidazole[4,5-d]pyrazolo[2,3-b]pyridinyl, thiazinyl, oxazinyl, thiadiazinyl, oxadiazinyl, dithiazinyl, dioxazinyl, oxathiazinyl, thiatriazinyl oxazinyl, oxatriazinyl, dithiadiazinyl, imidazolinyl, dihydropyrimidinyl, tetrahydropyrimidinyl, 1-pyrrolinyl, 2-pyrrolinyl, 3-pyrrolinyl, indolinyl, thiopyranyl, 2H-pyranyl, 4H-pyranyl, dioxanyl, 1,3-dioxolane, pyrazolinyl, pyrazolidinyl, dithianyl, dithiolanyl, pyrimidinone, pyrimidinedione, pyrimidine-2,4-dione, piperazinone, piperazinedionyl, pyrazolidinimidazolinyl, 3-azabicyclo[3.1.0]hexanyl, 3,6-diazabicyclo[3.1.1]heptanyl, 6-azabicyclo[3.1.1]heptanyl, 3-aza bicyclo[3.1.1]heptanyl, 3-azabicyclo[4.1.0]heptanyl, azabicyclo[2.2.2]hexanyl, 2-azabicyclo[3.2.1]octanyl, 8-azabicyclo[3.2.1]octanyl, 2-azabicyclo[2.2.2]octanyl, 8-azabicyclo[2.2.2]octanyl, 7-oxabicyclo[2.2.1]heptane, azaspiro[3.5]nonanyl, azaspiro[2.5]octanyl, azaspiro[4.5]decanyl, 1-azaspiro[4.5]decanyl, azaspiro[5.5]undecyl, tetrahydroindolyl, octahydroindolyl, tetrahydroisoindolyl, tetrahydroindazolyl, 1,1-dioxohexahydrothiopyranyl. Examples of 5-membered heterocyclic groups containing sulfur or oxygen atoms and one to three nitrogen atoms include thiazolyl, including thiazol-2-yl and thiazol-2-yl N-oxide; thiadiazolyl, including 1,3,4-thiadiazol-5-yl and 1,2,4-thiadiazol-5-yl; oxazolyl, such as oxazol-2-yl; and oxadiazolyl, such as 1,3,4-oxadiazol-5-yl and 1,2,4-oxadiazol-5-yl. Examples of 5-membered heterocyclic groups containing 2 to 4 nitrogen atoms include imidazolyl, such as imidazol-2-yl; triazolyl, such as 1,3,4-triazol-5-yl; 1,2,3-triazol-5-yl, 1,2,4-triazol-5-yl; and tetrazolyl, such as 1H-tetrazol-5-yl. Examples of benzo-fused 5-membered heterocyclic groups are benzoxazol-2-yl, benzothiazol-2-yl, and benzimidazol-2-yl. Exemplary 6-membered heterocyclic groups contain one to three nitrogen atoms and optionally sulfur or oxygen atoms, for example, pyridyl, such as pyridin-3-yl and pyridin-4-yl; pyrimidinyl, such as pyrimidin-2-yl and pyrimidin-4-yl; triazinyl, such as 1,3,4-triazin-2-yl and 1,3,5-triazin-4-yl; pyridazinyl, specifically pyridazin-3-yl; and pyrazinyl. Other exemplary heterocyclic groups are pyridine N-oxide and pyridazine N-oxide, as well as pyridyl, pyrimidin-2-yl, pyrimidin-4-yl, pyridazinyl, and 1,3,4-triazin-2-yl.

As used herein, the term "partially unsaturated" refers to a ring moiety that contains at least one double or triple bond between ring atoms but the ring moiety is not aromatic.

As used herein, the term "inhibitor" refers to a compound that binds to and inhibits PCAF with measurable affinity and activity. In certain embodiments, the inhibitor has an _IC50 or binding constant of less than about 20 μM, less than about 1 μM, less than about 500 nM, less than about 100 nM, or less than about 10 nM.

As used herein, the terms "measurable affinity" and "measurable inhibition" refer to a measurable decrease in the activity of a bromodomain PCAF (e.g., reduced recognition of lysine acetyl-recognizing chromatin) between (i) a sample comprising a compound of Formula I or a combination thereof and such a bromodomain, and (ii) an equivalent sample or composition thereof comprising such a bromodomain in the absence of the compound.

"Pharmaceutically acceptable salts" include acid and base addition salts. It should be understood that when a compound or example herein is shown as a particular salt, it is intended to encompass the corresponding free base as well as other salts of the corresponding free base (including pharmaceutically acceptable salts of the corresponding free base).

"Pharmaceutically acceptable acid addition salts" refers to those salts formed with inorganic or organic acids that retain the biological effectiveness and properties of the free bases and do not possess biologically or other undesirable effects and properties, such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, carbonic acid, phosphoric acid, and the like; and organic acids that can be selected from aliphatic, cycloaliphatic, aromatic, araliphatic, heterocyclic, carboxylic and sulfonic acid classes, such as formic acid, acetic acid, propionic acid, glycolic acid, gluconic acid, lactic acid, pyruvic acid, oxalic acid, malic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, aspartic acid, ascorbic acid, glutamic acid, anthranilic acid, benzoic acid, cinnamic acid, mandelic acid, pamoic acid, phenylacetic acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, salicylic acid, and the like.

"Pharmaceutically acceptable base addition salts" include those derived from inorganic bases such as sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, aluminum salts. Specific base addition salts are ammonium, potassium, sodium, calcium, and magnesium salts. Salts derived from pharmaceutically acceptable organic non-toxic bases include salts of primary, secondary, and tertiary amines; substituted amines, including naturally occurring substituted amines, cyclic amines, and basic ion exchange resins such as isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, ethanolamine, 2-diethylaminoethanol, tromethamine, dicyclohexylamine, lysine, arginine, histidine, caffeine, procaine, hydrazine, choline, betaine, ethylenediamine, glucosamine, methylglucamine, theobromine, purines, piperazine, piperidine, N-ethylpiperidine, polyamine resins, and the like. Specific organic non-toxic bases are isopropylamine, diethylamine, ethanolamine, tromethamine, dicyclohexylamine, choline and caffeine.

The term "tautomer" or "tautomeric form" refers to structural isomers of different energies that interconvert via a low energy barrier. For example, proton tautomers (also known as prototropic tautomers) include interconversions via proton migration, such as keto-enol and imine-enamine isomerizations. Valence tautomers include interconversions by reorganization of some of the bonding electrons.

"Solvate" refers to an association or complex of one or more solvent molecules and a compound of the present invention. Examples of solvents include water, isopropanol, ethanol, methanol, DMSO, ethyl acetate, acetic acid, and ethanolamine. The term "hydrate" refers to a complex in which the solvent molecule is water.

"Therapeutically effective amount" refers to an amount of a compound of the present invention that (i) treats a specific disease, condition, or disorder, (ii) alleviates, improves, or eliminates one or more symptoms of a specific disease, condition, or disorder, or (iii) delays the onset of one or more symptoms of a specific disease, condition, or disorder as described herein. In the case of cancer, a therapeutically effective amount of a drug can reduce the number of cancer cells; reduce tumor size; inhibit (i.e., slow down and preferably stop to some extent) cancer cell infiltration into peripheral organs; inhibit (i.e., slow down and preferably stop to some extent) tumor metastasis; inhibit tumor growth to some extent; and/or alleviate one or more cancer-related symptoms to some extent. For cancer treatment, efficacy can be measured, for example, by assessing time to disease progression (TTP) and/or determining a response rate (RR). In the case of immunological disorders, a therapeutically effective amount is an amount sufficient to alleviate or relieve symptoms of allergic disorders, autoimmune and/or inflammatory diseases, or symptoms of acute inflammatory reactions (e.g., asthma).

Variations of "treatment" (and variations such as "treat" or "treating") refer to attempts to alter the natural course of the individual or cell being treated, and can be used for clinical interventions performed preventively or during clinical pathology. Desirable effects of treatment include one or more of the following: prevention of recurrence of the disease; alleviation of symptoms; alleviation of any direct or indirect pathological consequences of the disease; stabilization of the disease state (i.e., no worsening); prevention of metastasis; reduction in the rate of disease progression; improvement or alleviation of the disease state; prolongation of survival compared to that expected if not receiving treatment and alleviation or improved prognosis. In certain embodiments, the compounds of Formula I are used to delay the development of a disease or condition or to slow the progression of a disease or condition. Those in need of treatment include those patients already suffering from the condition or condition, as well as those susceptible to the condition or condition (e.g., by genetic mutation or abnormal expression of a gene or protein).

"PCAF bromodomain inhibitors" refers to compounds that bind to the PCAF bromodomain and inhibit and/or reduce the biological activity of PCAF. In some embodiments, a PCAF bromodomain inhibitor binds to PCAF primarily through contact and/or interaction (e.g., solely) with the PCAF bromodomain. In some embodiments, a PCAF bromodomain inhibitor substantially or completely inhibits the biological activity of PCAF. In some embodiments, the biological activity is binding of the PCAF bromodomain to chromatin (e.g., histones associated with DNA) and/or another acetylated protein.

As used herein, the terms "PCAF" and "P300/CBP-associated factor," "K (lysine) acetyltransferase 2B," "KAT2B," "histone acetylase PCAF," "histone acetyltransferase PCAF," "lysine acetyltransferase 2B," "P/CAF," "EC 2.3.1.48," "CAF," "CREBBP-associated factor," and "histone acetyltransferase KAT2B" are used interchangeably and refer to any native PCAF from any vertebrate source, including mammals such as primates (e.g., humans) and rodents (e.g., mice and rats), unless otherwise indicated. The term encompasses "full-length" unprocessed PCAF as well as any form of PCAF resulting from processing in cells. The term also includes naturally occurring variants of PCAF, such as splice variants or allelic variants.

The term GCN5 includes GCN5L2, PCAF-b, KAT2A, STAF97, etc.

As used herein, "a" or "an" means one or more, unless expressly stated otherwise. As used herein, "another" means at least another or more.

Example variables

In one embodiment, the compound of formula (I) has a formula selected from formula I(a-j):

wherein Ring A is optionally substituted with one or more groups independently selected from the group consisting of C _1-6 alkyl, C _2-6 alkenyl, C _2-6 alkynyl, carbocyclyl, heterocyclyl, halogen, -NO ₂ , -N(R ^d ) ₂ , -CN, -C(O)-N(R ^d ) ₂ , -S(O)-N(R ^d ) ₂ , -S(O) ₂ -N(R ^d ) ₂ , -OR ^d , -SR ^d , -OC(O)-R ^d , -C(O)-R ^d , -C(O)-OR ^d , -S(O)-R ^d , -S(O) ₂ -R ^d , -C(O)-N(R ^d ) ₂ , -N(R ^d )-C(O)-R ^d , -N(R ^d )-S(O)-R d , ^d and -N(R ^d )-S(O) ₂ -R ^d , wherein any C _1-6 alkyl, C _2-6 alkenyl, C _2-6 alkynyl, carbocyclyl and heterocyclyl are optionally substituted with one or more groups independently selected from the group consisting of oxo, halogen, -NO ₂ , -N(R ^d ) ₂ , -CN, -C(O)-N(R ^d ) ₂ , -S(O)-N(R ^d ) ₂ , -S(O) ₂ -N(R ^d ) ₂ , -OR ^d , -SR ^d , -OC(O)-R ^d , -C(O)-R ^d , -C(O)-OR ^d , -S(O)-R ^d , -S(O) ₂ -R ^d , -C(O)-N(R ^d ) ₂ , -N(R ^d )-C(O)-R ^d , -N(R ^d )-S(O)-R ^d , -N(R ^d )-S(O) ₂ -R ^d ;

and its salts.

In another embodiment, Ring A is optionally substituted with one or more groups independently selected from the group consisting of C _1-6 alkyl, phenyl, halogen, and -OR ^d , wherein any of the C _1-6 alkyl and phenyl groups are optionally substituted with one or more groups independently selected from halogen.

In another embodiment, Ring A is optionally substituted with one or more groups independently selected from the group consisting of methoxy, 4-chlorophenyl, fluoro, and methyl.

In another embodiment, R ¹ is C _1-6 alkyl, optionally substituted with one or more groups independently selected from the group consisting of oxo, halogen, amino, hydroxy, and C _1-6 alkoxy.

In another embodiment, R ¹ is methyl or ethyl.

In another embodiment, X is O or S.

In another embodiment, X is N(H).

In another embodiment, R ³ is hydrogen or C _1-6 alkyl.

In another embodiment, R ³ is hydrogen, methyl or ethyl.

In another embodiment, R ⁴ is hydrogen, C _1-6 alkyl, carbocyclyl, or heterocyclyl, wherein each C _1-6 alkyl, carbocyclyl, and heterocyclyl is optionally substituted with one or more groups R ^b .

In another embodiment, ^R4 is selected from H, phenyl, methyl, ethyl, cyclohexyl, 4-fluorophenyl, 4-methoxyphenyl, 4-chlorophenyl, 1-methyltriazol-4-yl, 1-phenylpyrazol-4-yl, 1-methylpyrazol-4-yl, pyridin-3-yl, 4-(phenylsulfonyl)phenyl, 4-hydroxyphenyl, 4-(methoxycarbonyl)phenyl, 2-chlorophenyl, 2-fluorophenyl, 4-carboxyphenyl,

In another embodiment, R ³ and R ⁴ together with the atoms to which they are attached form a carbocyclyl or heterocyclyl group, which is optionally substituted with one or more groups R ^b .

In another embodiment, R ³ and R ⁴ together with the atoms to which they are attached form a carbocyclic or heterocyclic group selected from a cyclohexane group, a cyclopentane group, a piperidine group and an indan group, said carbocyclic or heterocyclic group being optionally substituted with one or more groups R ^b .

In another embodiment, one R ^e and R ³ together with the atoms to which they are attached form a heterocyclyl; optionally substituted with one or more groups independently selected from the group consisting of C _1-6 alkyl, C _2-6 alkenyl, C _2-6 alkynyl and C _1-6 alkoxy, wherein each C _1-6 alkyl, C _2-6 alkenyl, C _2-6 alkynyl and C _1-6 alkoxy is optionally substituted with one or more groups independently selected from the group consisting of oxo, halogen, amino, hydroxy, C _1-6 alkoxy and carbocyclyl.

In another embodiment, one R ^e and R ³ together with the atoms to which they are attached form an azetidine ring, said ring optionally substituted with one or more groups independently selected from the group consisting of C _1-6 alkyl, C _2-6 alkenyl, C _2-6 alkynyl and C _1-6 alkoxy, wherein each C _1-6 alkyl, C _2-6 alkenyl, C _2-6 alkynyl and C _1-6 alkoxy is optionally substituted with one or more groups independently selected from the group consisting of oxo, halogen, amino, hydroxy, C _1-6 alkoxy and carbocyclyl.

In another embodiment, one R ^e and R ⁴ together with the atoms to which they are attached form a heterocyclyl; optionally substituted with one or more groups independently selected from the group consisting of C _1-6 alkyl, C _2-6 alkenyl, C _2-6 alkynyl and C _1-6 alkoxy, wherein each C _1-6 alkyl, C _2-6 alkenyl, C _2-6 alkynyl and C _1-6 alkoxy is optionally substituted with one or more groups independently selected from the group consisting of oxo, halogen, amino, hydroxy, C _1-6 alkoxy and carbocyclyl.

In another embodiment, one R ^e and R ⁴ together with the atoms to which they are attached form a heterocyclic group selected from a pyrrolidino group, a piperidino group and an azepanyl group, said heterocyclic group being optionally substituted with one or more groups independently selected from the group consisting of C _1-6 alkyl, C _2-6 alkenyl, C _2-6 alkynyl and C _1-6 alkoxy, wherein each C _1-6 alkyl, C _2-6 alkenyl, C _2-6 alkynyl and C _1-6 alkoxy is optionally substituted with one or more groups independently selected from oxo, halogen, amino, hydroxy, C _1-6 alkoxy and carbocyclyl.

In another embodiment, the group:

Selected from:

In another embodiment, the group:

Selected from:

In another embodiment, the group:

Selected from:

In another embodiment, the compound is selected from:

and its salts.

In certain embodiments, the compound of formula (I) is a mixture as described in the Examples herein, or a free base or salt thereof.

Use, formulation and administration

Pharmaceutically acceptable compositions

Another aspect includes a pharmaceutical composition comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof. In one embodiment, the composition further comprises a pharmaceutically acceptable carrier, adjuvant, or vehicle. In another embodiment, the composition further comprises an amount of a compound effective to measurably inhibit the bromodomain of PCAF. In certain embodiments, the composition is formulated for administration to a patient in need thereof.

As used herein, the term "patient" or "individual" refers to an animal, such as a mammal, such as a human. In one embodiment, the patient or individual is a human.

The term "pharmaceutically acceptable carrier, adjuvant or vehicle" refers to a non-toxic carrier, adjuvant or vehicle that does not destroy the pharmacological activity of the compound formulated therewith. Pharmaceutically acceptable carriers, adjuvants or vehicles that can be used in the compositions of the present invention include, but are not limited to, ion exchangers, aluminum oxide, aluminum stearate, lecithin, serum proteins such as human serum albumin, buffer substances such as phosphates, glycine, sorbic acid, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silicon dioxide, magnesium trisilicate, polyvinyl pyrrolidone, cellulose-based substances, polyethylene glycol, sodium carboxymethyl cellulose, polyacrylates, waxes, polyethylene-polyoxypropylene block polymers, polyethylene glycol and lanolin.

The compositions comprising a compound of Formula I or a salt thereof can be administered orally, parenterally, by inhalation spray, topically, transdermally, rectally, nasally, buccally, sublingually, vaginally, intraperitoneally, intrapulmonary, intradermally, epidurally or via an implanted reservoir. The term "parenteral" as used herein includes subcutaneous, intravenous, intramuscular, intraarticular, intrasynovial, intrasternal, intrathecal, intrahepatic, intracerebral and intracranial injection or infusion techniques.

In one embodiment, the composition comprising a compound of Formula I or a salt thereof is formulated into a solid dosage form for oral administration. Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In certain embodiments, a solid oral dosage form of a compound of formula (I) or a salt thereof further comprises one or more of the following: (i) an inert, pharmaceutically acceptable excipient or carrier, such as sodium citrate or dicalcium phosphate; and (ii) a filler or extender, such as starch, lactose, sucrose, glucose, mannitol, or silicic acid; (iii) a binder, such as carboxymethylcellulose, an alginate, gelatin, polyvinylpyrrolidone, sucrose, or acacia; (iv) a humectant, such as glycerol; (v) a disintegrant, such as agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates, or sodium carbonate; (vi) a solvent blocker, such as paraffin; (vii) an absorption enhancer, such as a quaternary ammonium salt; (viii) a wetting agent, such as cetyl alcohol or glyceryl monostearate; (ix) an absorbent, such as kaolin or bentonite; and (x) a lubricant, such as talc, calcium stearate, magnesium stearate, polyethylene glycol, or sodium lauryl sulfate. In certain embodiments, the solid oral dosage form is formulated into a capsule, tablet or pill. In certain embodiments, the solid oral dosage form further comprises a buffer. In certain embodiments, such compositions for solid oral dosage forms can be formulated into fillers in soft and hard filled gelatin capsules comprising one or more excipients such as lactose or milk sugar, polyethylene glycol, etc.

In certain embodiments, tablets, dragees, capsules, pills and granules comprising a composition of a compound of Formula I or a salt thereof optionally comprise a coating or shell, such as an enteric coating, which may optionally comprise an opacifying agent, and may also be in the form of a composition that releases the active ingredient only in or preferably in a certain portion of the intestinal tract, optionally in a delayed manner. Examples of embedding compositions include polymeric substances and waxes, which may also use excipients such as lactose or milk sugar and high molecular weight polyethylene glycols as fillers in soft and hard-filled gelatin capsules.

In another embodiment, the composition comprises a microencapsulated compound of (I) or a salt thereof, and optionally further comprises one or more excipients.

In another embodiment, the composition comprises a liquid dosage formulation containing a compound of formula I or a salt thereof for oral administration, and optionally further comprises one or more pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups and elixirs. In certain embodiments, the liquid dosage form optionally further comprises one or more inert diluents such as water or other solvents; solubilizers and emulsifiers such as ethanol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, dimethylformamide, oil (specifically cottonseed oil, peanut oil, corn oil, germ oil, olive oil, castor oil and sesame oil), glycerol, tetrahydrofurfuryl alcohol, polyethylene glycol or fatty acid esters of dehydrated sorbitol and mixtures thereof. In certain embodiments, the liquid oral composition optionally further comprises one or more adjuvants, such as wetting agents, suspending agents, sweeteners, flavoring agents and aromatics.

Injectable preparations, such as sterile injectable aqueous or oily suspensions, can be prepared using suitable dispersants or wetting agents and suspending agents according to known techniques. Sterile injectable preparations can also be sterile injectable solutions, suspensions or emulsions in nontoxic parenteral acceptable diluents or solvents, for example as solutions in 1,3-butanediol. Acceptable vehicles and solvents that can be used are water, Ringer's solution, U.S.P. and isotonic sodium chloride solution. In addition, sterile fixed oils are generally used as solvents or suspending media. For this purpose, any mild fixed oil can be used, including synthetic monoglycerides or diglycerides. In addition, fatty acids such as oleic acid can be used to prepare injections.

The injectable formulations can be sterilized, for example, by filtration through a bacteria-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.

In order to prolong the effect of a compound of formula (I), it is generally desirable to slow down the absorption of the compound from subcutaneous or intramuscular injection. This can be achieved by using a liquid suspension of crystalline or amorphous material with poor water solubility. The absorption rate of the compound then depends on its dissolution rate, which in turn depends on crystal size and crystalline form. Alternatively, the delayed absorption of the compound form administered parenterally is achieved by dissolving or suspending the compound in an oil vehicle. Injectable reservoir forms are prepared by forming a microcapsule matrix of the compound in a biodegradable polymer such as polylactide-polyglycolide. Depending on the ratio of the compound to the polymer and the properties of the specific polymer used, the rate of compound release can be controlled. Examples of other biodegradable polymers include poly(orthoesters) and poly(anhydrides). Depot injection formulations are also prepared by embedding the compound in liposomes or microemulsions compatible with body tissues.

In certain embodiments, compositions for rectal or vaginal administration are formulated as suppositories, which can be prepared by mixing a compound of formula (I) or a salt thereof with a suitable non-irritating excipient or carrier such as cocoa butter, polyethylene glycol, or a suppository wax, e.g., those which are solid at ambient temperature but liquid at body temperature and therefore melt in the rectum or vaginal cavity and release the compound of formula (I).

Examples of dosage forms for topical or transdermal administration of compound of formula (I) include ointments, pastes, creams, lotions, gels, powders, solutions, sprays, inhalants or patches. Compound of formula (I) or its salt is mixed with a pharmaceutically acceptable carrier, and an optional preservative or buffer under aseptic conditions. Other examples of formulations include ophthalmic preparations, ear drops, eye drops, transdermal patches. Transdermal formulations can be prepared as follows: compound of formula (I) or its salt is dissolved or distributed in a medium, such as ethanol or dimethyl sulfoxide. Absorption enhancers can also be used to increase the flux of the compound through the skin. Rate can be controlled by providing a rate-controlling membrane or by dispersing the compound in a polymer matrix or a gel.

Nasal aerosol or inhalation formulations of the compound of formula (I) or its salt may be prepared as saline solutions using benzyl alcohol or other suitable preservatives, absorption promoters for enhancing bioavailability, fluorocarbons and/or other conventional solubilizers or dispersants.

In certain embodiments, the pharmaceutical composition may be administered with or without food. In certain embodiments, the pharmaceutically acceptable composition is not administered with food. In certain embodiments, the pharmaceutically acceptable composition of the present invention is administered with food.

The specific dosage and treatment regimen for any particular patient will depend on a variety of factors, including age, weight, general health, sex, diet, time of administration, rate of excretion, drug combinations, the judgment of the treating physician, and the severity of the specific disease being treated. The amount of the compound of Formula I or its salt provided in the composition will also depend on the specific compound in the composition.

In one embodiment, the therapeutically effective amount of the compound of the invention administered parenterally per dose will be in the range of about 0.01-100 mg/kg patient body weight, or about 0.1-20 mg/kg patient body weight per day, with a typical initial range of 0.3 to 15 mg/kg/day of the compound used. In another embodiment, oral unit dosage forms such as tablets and capsules contain from about 5 to about 100 mg of the compound of the invention.

For example, a tablet oral dosage form comprises about 2 mg, 5 mg, 25 mg, 50 mg, 100 mg, 250 mg or 500 mg of a compound of formula (I) or a salt thereof, and further comprises about 5-30 mg of anhydrous lactose, about 5-40 mg of croscarmellose sodium, about 5-30 mg of polyvinylpyrrolidone (PVP) K30 and about 1-10 mg of magnesium stearate. The method of preparing the tablet comprises mixing the powdered ingredients together and further mixing with a PVP solution. The resulting composition can be dried, granulated, mixed with magnesium stearate, and compressed into tablet form using conventional equipment. An example of an aerosol formulation can be prepared as follows: about 2-500 mg of a compound of formula I or a salt thereof is dissolved in a suitable buffer solution, such as a phosphate buffer, and (if necessary) a thickening agent, such as a salt such as sodium chloride, is added. The solution can be filtered, for example using a 0.2 micron filter, to remove impurities and contaminants.

Uses of compounds and pharmaceutically acceptable compositions

Another aspect includes the use of a compound of formula (I) or a salt thereof for inhibiting (in vitro or in vivo) the bromodomain of PCAF.

Another embodiment includes a method of treating a PCAF-mediated disorder in an animal (eg, a mammal such as a human) comprising administering to the animal a compound of formula (I) or a pharmaceutically acceptable salt thereof. PCAF-mediated disorders include, but are not limited to, those described herein.

Another aspect includes the use of a compound of (I) or a salt thereof for inhibiting (in vitro or in vivo) the bromodomain of GCN5.

Another embodiment includes a method of treating a GCN5-mediated disorder in an animal (eg, a mammal such as a human) comprising administering to the animal a compound of Formula (I) or a pharmaceutically acceptable salt thereof.

Another embodiment includes a method of increasing the efficacy of a cancer therapy comprising a cytotoxic agent in an animal (eg, a mammal such as a human) comprising administering to the animal a compound of Formula (I) or a pharmaceutically acceptable salt thereof.

Another embodiment includes a method of delaying or preventing the development of resistance of a cancer to a cytotoxic agent in an animal (eg, a mammal such as a human) comprising administering to the animal a compound of Formula (I) or a pharmaceutically acceptable salt thereof.

Another embodiment includes a method of extending the duration of response to a cancer therapy in an animal (e.g., a mammal such as a human), comprising administering a compound of Formula (I) or a pharmaceutically acceptable salt thereof to an animal undergoing cancer therapy, wherein the duration of response to the cancer therapy is extended when the compound of Formula (I) or a pharmaceutically acceptable salt thereof is administered, compared to the duration of response to the cancer therapy in the absence of administration of the compound of Formula (I) or a pharmaceutically acceptable salt thereof.

Another embodiment includes a method of treating cancer in an individual, comprising administering to the individual (a) a compound of formula (I) or a pharmaceutically acceptable salt thereof, and (b) a cytotoxic agent. In one embodiment, the cytotoxic agent is selected from antimicrotubule agents, platinum coordination complexes, alkylating agents, antibiotics, topoisomerase II inhibitors, antimetabolites, topoisomerase I inhibitors, hormones and hormone analogs, signal transduction pathway inhibitors, non-receptor tyrosine kinase angiogenesis inhibitors, immunotherapeutics, pro-apoptotic agents, LDH-A inhibitors, fatty acid biosynthesis inhibitors, cell cycle signaling inhibitors, HDAC inhibitors, proteasome inhibitors, and cancer metabolism inhibitors. In one embodiment, the cytotoxic agent is a taxane. In one embodiment, the taxane is paclitaxel or docetaxel. In one embodiment, the cytotoxic agent is a platinum agent. In one embodiment, the cytotoxic agent is an antagonist of EGFR. In one embodiment, the EGFR antagonist is N-(3-ethynylphenyl)-6,7-bis(2-methoxyethoxy)quinazolin-4-amine or a pharmaceutically acceptable salt thereof (e.g., erlotinib). In one embodiment, the cytotoxic agent is a RAF inhibitor. In one embodiment, the RAF inhibitor is a BRAF or CRAF inhibitor. In one embodiment, the RAF inhibitor is vemurafenib. In one embodiment, the cytotoxic agent is a PI3K inhibitor.

In certain embodiments, treatment may be administered after one or more symptoms develop. In other embodiments, treatment may be administered in the absence of symptoms. For example, treatment may be administered to susceptible individuals prior to the onset of symptoms (e.g., in view of a history of symptoms and/or in view of genetic or other predisposing factors). Treatment may also be continued after symptoms resolve, e.g., to prevent or delay recurrence.

PCAF-mediated disorders

A "PCAF-mediated disorder" is characterized by the involvement of PCAF in the onset, manifestation, severity, or progression of the disorder, or one or more symptoms or disease markers.

PCAF-mediated disorders include cancers including, but not limited to, acoustic neuroma, acute leukemia, acute lymphocytic leukemia, acute myeloid leukemia, acute T-cell leukemia, androgen-responsive prostate cancer, basal cell carcinoma, bile duct cancer, bladder cancer, brain cancer, breast cancer, bronchial cancer, cervical cancer, chondrosarcoma, chordoma, choriocarcinoma, chronic leukemia, chronic lymphocytic leukemia, chronic myeloid leukemia, chronic myeloid leukemia, colon cancer, colorectal cancer, craniocarcinoma, ovarian cancer, ovarian cancer, ovarian cancer, ureteral ... Pharyngioma, cystadenocarcinoma, diffuse large B-cell lymphoma, drug-resistant breast cancer, dysproliferative changes, embryonal carcinoma, endometrial cancer, endothelial sarcoma, ependymoma, epithelial carcinoma, erythroleukemia, esophageal cancer, estrogen receptor-positive breast cancer, essential thrombocythemia, Ewing's tumor, fibrosarcoma, follicular lymphoma, gastric cancer, germ cell testicular cancer, glioma, glioblastoma, gliosarcoma, heavy chain disease, head and neck cancer, hemangioblastoma, liver cancer, hepatocellular carcinoma, hormone-insensitive prostate carcinoma, leiomyosarcoma, leukemia, liposarcoma, lung cancer, lymphangioendothelioma, lymphangiosarcoma, lymphocytic leukemia, lymphoma, lymphoid malignancies of T-cell or B-cell origin, medullary carcinoma, medulloblastoma, melanoma, meningioma, mesothelioma, multiple myeloma, myeloid leukemia, myeloma, myxosarcoma, neuroblastoma, NUT midline carcinoma (NMC), non-small cell lung cancer, oligodendroglioma, oral cancer, osteosarcoma, ovarian cancer, pancreatic cancer, papillary gland carcinoma, papillary carcinoma, pediatric acute lymphoblastic leukemia, pinealoma, polycythemia vera, prostate cancer, rectal cancer, renal cell carcinoma, retinoblastoma, rhabdomyosarcoma, sarcoma, sebaceous gland carcinoma, seminoma, skin cancer, small cell lung cancer, solid tumors (carcinomas and sarcomas), small cell lung cancer, gastric cancer, squamous cell carcinoma, synovioma, sweat gland carcinoma, thyroid cancer, Waldenstrom's macroglobulinemia, WNT-dependent breast cancer, testicular tumors, uterine cancer, and Wilms' tumor.

In certain embodiments, the cancer is selected from gastric cancer, lung cancer, non-small cell lung cancer, pancreatic cancer, pediatric acute lymphoblastic leukemia, androgen-responsive prostate cancer, breast cancer, Wnt-dependent breast cancer, drug-resistant breast cancer, estrogen receptor-positive breast cancer, leukemia, neuroblastoma, colon cancer, and cervical cancer. In certain embodiments, the cancer is lung cancer. In certain embodiments, the cancer is pancreatic cancer. In certain embodiments, the cancer is breast cancer. In certain embodiments, the cancer is leukemia. In certain embodiments, the cancer is colon cancer. In certain embodiments, the cancer is cervical cancer.

PCAF-mediated disorders also include inflammatory diseases, inflammatory conditions, and autoimmune diseases, including but not limited to: Addison's disease, acute gout, Alzheimer's disease (inflammatory-mediated neurotoxicity), ankylosing spondylitis, asthma, atherosclerosis, Behcet's disease, bullous skin diseases, chronic obstructive pulmonary disease (COPD), Crohn's disease, dermatitis, eczema, giant cell arteritis, glomerulonephritis, hepatitis, hypophysitis, inflammatory bowel disease, Kawasaki disease, lupus erythematosus, leukopenia ... Nephritis, multiple sclerosis, myocarditis, myositis, nephritis, organ transplant rejection, osteoarthritis, pancreatitis, pericarditis, polyarteritis nodosa, pneumonia, primary biliary cirrhosis, psoriasis, psoriatic arthritis, rheumatoid arthritis, scleritis, sclerosing cholangitis, sepsis, systemic lupus erythematosus, polyarteritis, toxic shock, thyroiditis, type I diabetes, ulcerative colitis, uveitis, vitiligo, vasculitis, and Wegener's granulomatosis. In some embodiments, the condition is an autoimmune disease. In some embodiments, the condition is asthma.

PCAF-mediated conditions also include AIDS; chronic kidney disease, including but not limited to diabetic nephropathy, hypertensive nephropathy, HIV-associated nephropathy, glomerulonephritis, lupus nephritis, IgA nephropathy, focal segmental glomerulosclerosis, membranous glomerulonephritis, minimal change disease, polycystic kidney disease, and tubulointerstitial nephritis; acute kidney injury or disease or condition, including but not limited to ischemia-reperfusion induced, cardiac and major surgery induced, percutaneous coronary intervention induced, radiocontrast agent induced, sepsis induced, pneumonia induced, and drug toxicity induced acute kidney injury or disease or condition; HIV infection; obesity; osteoporosis, dyslipidemia; hypercholesterolemia; Alzheimer's disease; metabolic syndrome; hepatic steatosis; type II diabetes; insulin resistance; diabetic retinopathy; osteoporosis; obesity, and parasitic infection (e.g., Toxoplasma gondii). In some embodiments, the condition is osteoporosis. In some embodiments, the condition is obesity. In some embodiments, the condition is HIV infection. In some embodiments, the condition is a parasitic infection.

GCN5-mediated disorders

A "GCN5-mediated disorder" is characterized by the involvement of GCN5 in the onset, manifestation of one or more symptoms or disease markers, severity, or progression of the disorder.

GCN5-mediated disorders include cancers including, but not limited to, acoustic neuroma, acute leukemia, acute lymphocytic leukemia, acute myeloid leukemia, acute T-cell leukemia, androgen-responsive prostate cancer, basal cell carcinoma, bile duct cancer, bladder cancer, brain cancer, breast cancer, bronchial cancer, cervical cancer, chondrosarcoma, chordoma, choriocarcinoma, chronic leukemia, chronic lymphocytic leukemia, chronic myeloid leukemia, chronic myeloid leukemia, colon cancer, colorectal cancer, craniocarcinoma, ovarian cancer, ovarian cancer, ovarian cancer, ureteral cancer, ureteral cancer, urothelial carcinoma, urothelial carcinoma, urothelial carcinoma, urothelial carcinoma, ureteral cancer ... Pharyngioma, cystadenocarcinoma, diffuse large B-cell lymphoma, drug-resistant breast cancer, dysproliferative changes, embryonal carcinoma, endometrial cancer, endothelial sarcoma, ependymoma, epithelial carcinoma, erythroleukemia, esophageal cancer, estrogen receptor-positive breast cancer, essential thrombocythemia, Ewing's tumor, fibrosarcoma, follicular lymphoma, gastric cancer, germ cell testicular cancer, glioma, glioblastoma, gliosarcoma, heavy chain disease, head and neck cancer, hemangioblastoma, liver cancer, hepatocellular carcinoma, hormone-insensitive prostate carcinoma, leiomyosarcoma, leukemia, liposarcoma, lung cancer, lymphangioendothelioma, lymphangiosarcoma, lymphocytic leukemia, lymphoma, lymphoid malignancies of T-cell or B-cell origin, medullary carcinoma, medulloblastoma, melanoma, meningioma, mesothelioma, multiple myeloma, myeloid leukemia, myeloma, myxosarcoma, neuroblastoma, NUT midline carcinoma (NMC), non-small cell lung cancer, oligodendroglioma, oral cancer, osteosarcoma, ovarian cancer, pancreatic cancer, papillary gland carcinoma, papillary carcinoma, pediatric acute lymphoblastic leukemia, pinealoma, polycythemia vera, prostate cancer, rectal cancer, renal cell carcinoma, retinoblastoma, rhabdomyosarcoma, sarcoma, sebaceous gland carcinoma, seminoma, skin cancer, small cell lung cancer, solid tumors (carcinomas and sarcomas), small cell lung cancer, gastric cancer, squamous cell carcinoma, synovioma, sweat gland carcinoma, thyroid cancer, Waldenstrom's macroglobulinemia, WNT-dependent breast cancer, testicular tumors, uterine cancer, and Wilms' tumor.

In certain embodiments, the cancer is selected from gastric cancer, lung cancer, non-small cell lung cancer, pancreatic cancer, pediatric acute lymphoblastic leukemia, androgen-responsive prostate cancer, breast cancer, Wnt-dependent breast cancer, drug-resistant breast cancer, estrogen receptor-positive breast cancer, leukemia, neuroblastoma, colon cancer, and cervical cancer. In certain embodiments, the cancer is lung cancer. In certain embodiments, the cancer is pancreatic cancer. In certain embodiments, the cancer is breast cancer. In certain embodiments, the cancer is leukemia. In certain embodiments, the cancer is colon cancer. In certain embodiments, the cancer is cervical cancer.

GCN5-mediated disorders also include inflammatory diseases, inflammatory conditions, and autoimmune diseases, including but not limited to: Addison's disease, acute gout, Alzheimer's disease (inflammatory-mediated neurotoxicity), ankylosing spondylitis, asthma, atherosclerosis, Behcet's disease, bullous skin diseases, chronic obstructive pulmonary disease (COPD), Crohn's disease, dermatitis, eczema, giant cell arteritis, glomerulonephritis, hepatitis, hypophysitis, inflammatory bowel disease, Kawasaki disease, lupus erythematosus, leukemia, leukopenia ... Nephritis, multiple sclerosis, myocarditis, myositis, nephritis, organ transplant rejection, osteoarthritis, pancreatitis, pericarditis, polyarteritis nodosa, pneumonia, primary biliary cirrhosis, psoriasis, psoriatic arthritis, rheumatoid arthritis, scleritis, sclerosing cholangitis, sepsis, systemic lupus erythematosus, polyarteritis, toxic shock, thyroiditis, type I diabetes, ulcerative colitis, uveitis, vitiligo, vasculitis, and Wegener's granulomatosis. In some embodiments, the condition is an autoimmune disease. In some embodiments, the condition is asthma.

GCN5-mediated conditions also include AIDS; chronic kidney disease, including but not limited to diabetic nephropathy, hypertensive nephropathy, HIV-associated nephropathy, glomerulonephritis, lupus nephritis, IgA nephropathy, focal segmental glomerulosclerosis, membranous glomerulonephritis, minimal change disease, polycystic kidney disease, and tubulointerstitial nephritis; acute kidney injury or disease or condition, including but not limited to ischemia-reperfusion induced, cardiac and major surgery induced, percutaneous coronary intervention induced, radiocontrast agent induced, sepsis induced, pneumonia induced, and drug toxicity induced acute kidney injury or disease or condition; HIV infection; obesity; osteoporosis, dyslipidemia; hypercholesterolemia; Alzheimer's disease; metabolic syndrome; hepatic steatosis; type II diabetes; insulin resistance; diabetic retinopathy; osteoporosis; obesity and parasitic infection (e.g., Toxoplasma gondii). In some embodiments, the condition is osteoporosis. In some embodiments, the condition is obesity. In some embodiments, the condition is HIV infection. In some embodiments, the condition is a parasitic infection.

Co-administration of compounds and other pharmaceutical agents

The compound of formula (I) or its salt can be used alone or in combination with other drugs. For example, the second agent of the drug combination formulation or dosing regimen may have an activity complementary to the compound of formula (I) so that they do not adversely affect each other. The compound can be administered together in an integrated pharmaceutical composition or separately. In one embodiment, the compound or pharmaceutically acceptable salt can be co-administered with a cytotoxic agent to treat proliferative diseases and cancer.

The term "co-administration" refers to the simultaneous administration or separate sequential administration of a compound of formula (I) or a salt thereof and one or more other active pharmaceutical ingredients, including cytotoxic agents and radiation therapy, in any manner. If not administered simultaneously, the compounds are administered close in time to each other. In addition, it is not important whether the compounds are administered in the same dosage form, for example, one compound may be administered topically and the other compound may be administered orally.

As part of a multiple-dose regimen, these additional agents may be administered separately from the composition containing the compounds of the invention. Alternatively, these agents may be part of a single dosage form, mixed together with the compounds of the invention in a single composition. If administered as part of a multiple-dose regimen, the two active agents may be administered simultaneously, sequentially, or within a period of 5 hours of each other.

As used herein, the terms "combination," "combination," and related terms refer to the simultaneous or sequential administration of therapeutic agents according to the present invention. For example, a compound of the present invention can be administered with another therapeutic agent simultaneously or sequentially in separate unit dosage forms or in a single dosage form. Thus, the present invention provides a single unit dosage form comprising a compound of Formula I, an additional therapeutic agent, and a pharmaceutically acceptable carrier, adjuvant, or vehicle.

The amount of both the compound of the invention and the additional therapeutic agent (in those compositions comprising additional therapeutic agents as described above) that can be combined with the carrier materials to produce a single dosage form will vary depending on the host being treated and the specific mode of administration. In certain embodiments, the compositions of the invention are formulated so that a dosage of 0.01-100 mg/kg body weight/day of the invention can be administered.

In general, any agent that is active against the disease or condition being treated can be co-administered. Examples of such agents can be found in Cancer Principles and Practice of Oncology by VTDevita and S. Hellman (editors), ⁶ edition (February 15, 2001), Lippincott Williams & Wilkins Publishers. One of ordinary skill in the art will be able to discern which combinations of agents will be useful based on the specific characteristics of the drugs and diseases involved.

In one embodiment, the method of treatment includes co-administration of a compound of formula (I) or a pharmaceutically acceptable salt thereof and at least one cytotoxic agent. The term "cytotoxic agent" as used herein refers to a substance that inhibits or prevents cell function and/or causes cell death or destruction. Cytotoxic agents include, but are not limited to, radioisotopes (e.g., At ²¹¹ , I ¹³¹ , I ¹²⁵ , Y ⁹⁰ , Re ¹⁸⁶ , Re ¹⁸⁸ , Sm ¹⁵³ , Bi ²¹² , P ³² , Pb ²¹² and radioisotopes of Lu); chemotherapeutic agents; growth inhibitors; enzymes and fragments thereof such as nuclear decomposition enzymes; and toxins, such as small molecule toxins or enzymatically active toxins of bacterial, fungal, plant or animal origin, including fragments and/or variants thereof.

Exemplary cytotoxic agents can be selected from antimicrotubule agents, platinum coordination complexes, alkylating agents, antibiotics, topoisomerase II inhibitors, antimetabolites, topoisomerase I inhibitors, hormones and hormone analogs, signal transduction pathway inhibitors, non-receptor tyrosine kinase angiogenesis inhibitors, immunotherapeutic agents, pro-apoptotic agents, LDH-A inhibitors; fatty acid biosynthesis inhibitors; cell cycle signaling inhibitors; HDAC inhibitors, proteasome inhibitors; and cancer metabolism inhibitors.

"Chemotherapeutic agents" include compounds that can be used to treat cancer. Examples of chemotherapeutic agents include erlotinib (Genentech/OSI Pharm.), bortezomib (Millennium Pharm.), disulfiram, epigallocatechin gallate, salinosporamide A, carfilzomib, 17-AAG (geldanamycin), radicicol, lactate dehydrogenase A (LDH-A), fulvestrant (AstraZeneca), sunitinib (Pfizer/Sugen), letrozole (Novartis), imatinib mesylate (Novartis), finasunate (Novartis), oxaliplatin (Sanofi), 5-fluorouracil (5-FU), leucovorin, rapamycin (Sirolimus, Wyeth), lapatinib (GSK572016, GlaxoSmithKline), lonafamib (SCH 66336), sorafenib (Bayer Labs), gefitinib (AstraZeneca), AG1478, alkylating agents such as thiotepa and cyclophosphamide; alkyl sulfonates such as busulfan, improsulfan, and piposulfan; aziridines such as benzodopa, carboquone, meturedopa, and uredopa; ethylenimines and methylamelamines, including hexamethylmelamine ( altretamine), triethylenemelamine, triethylenephosphoramide, triethylenethiophosphoramide, and trimethylomelamine; acetogenins (especially bullatacin and bullatacinone); camptothecins (including topotecan and irinotecan); lichen inhibitors bryostatin; callystatin; CC-1065 (including its synthetic analogs adozelesin, carzelesin, and bizelesin); cryptophycins (particularly cryptophycin 1 and cryptophycin 8); adrenocortical steroids (including prednisone and prednisolone); cyproterone acetate; 5α-reductase inhibitors, including finasteride and dutasteride; vorinostat; inostat), romidepsin, panobinostat, valproic acid, mocetinostat, dolastatin; aldesleukin, talc, duocarmycin (including synthetic analogs KW-2189 and CB1-TM1); eleutherobin; pancratistatin; sarcodictyin; spongistatin; nitrogen mustards mustards, such as chlorambucil, chlornaphazine, chlorophosphamide, estramustine, ifosfamide, mechlorethamine, mechlorethamine oxide hydrochloride, melphalan, novembichin, phenesterine, prednimustine, trofosfamide, uracil mustard, mustard; nitrosureas, such as carmustine, chlorozotocin, fotemustine, lomustine, nimustine, and ranimustine; antibiotics, such as the enediyne antibiotics (e.g., calicheamicin, especially calicheamicin gamma and calicheamicin omega (Angew, Chem. Intl. Ed. Engl. 1994). 33:183-186); dynemicins, including dynemicin A; bisphosphonates, such as clodronate; esperamicin; and neocarzinostatin chromophores and related chromoprotein enediyne antibiotic chromophores), aclacinomycins, actinomycin, authramycin, azaserine, bleomycin, cactinomycin, carabicin, carminomycin , carzinophilin, chromomycin, dactinomycin, daunorubicin, detorubicin, 6-diazo-5-oxo-L-norleucine, (doxorubicin), morpholino-doxorubicin, cyanomorpholino-doxorubicin, 2-pyrrolino-doxorubicin and deoxydoxorubicin), epirubicin, esorubicin, idarubicin, marcellomycin, mitomycins such as mitomycin C, mycophenolic acid acid, nogalamycin, olivomycin, peplomycin, porfiromycin, puromycin, quelamycin, rodorubicin, streptonigrin, streptozocin, tubercidin, ubenimex, zinostatin, zorubicin; antimetabolites such as methotrexate and 5-fluorouracil (5-FU); folic acid analogs such as denopterin, methylprednisolone, Aminopterin, pteropterin, trimetrexate; purine analogs such as fludarabine, 6-mercaptopurine, thiamiprine, thioguanine; pyrimidine analogs such as ancitabine, azacitidine, 6-azauridine, carmofur, cytarabine, dideoxyuridine, doxifluridine, enocitabine, floxuridine; androgens such as calusterone, dromostanolone propionate propionate, epitiostanol, mepitiostane, testolactone; antiadrenal drugs such as aminoglutethimide, mitotane, and trilostane; folic acid supplements such as folinic acid; aceglatone; aldophosphamide glycoside; aminolevulinic acid; eniluracil; amsacrine; bestrabucil; bisantrene; edatraxate; defofamine; demecolcine; diaziquone; elfomithine (eflornithine); elliptinium acetate acetate; epothilone; etoglucid; gallium nitrate; hydroxyurea; lentinan; lonidamine; maytansinoids, such as maytansine and ansamitocin; mitoguazone; mitoxantrone; mopidamol; nitracrine; pentostatin; phenamet; pirarubicin; losoxantrone; podophyllinic acid; 2-ethylhydrazide; procarbazine; polysaccharide complex (JHS Natural Products, Eugene, Oreg.); razoxane; rhizoxin; sizofiran; spirogermanium; tenuazonic acid; triaziquone; 2,2',2"-trichlorotriethylamine; trichothecenes (especially T-2 toxin, verracurin A, roridin A, and anguidine); urethan; vindesine; dacarbazine; mannomustine; mitobronitol; mitolactol; pipobroman; gacytosine; arabinoside ("Ara-C"); cyclophosphamide; thiotepa; taxoids, such as TAXOL (paclitaxel; Bristol-Myers Squibb Oncology, Princeton, NJ), (without Cremophor), and albumin-engineered nanoparticle formulations of paclitaxel (American Pharmaceutical Partners, Schaumberg, Ill.) and (docetaxel (doxetaxel); Sanofi-Aventis), chloranmbucil; (gemcitabine); 6-thioguanine; mercaptopurine; methotrexate; platinum analogs such as cisplatin and carboplatin; vinblastine; etoposide (VP-16); ifosfamide; mitoxantrone; vincristine (vincris tine); (vinorelbine); novantrone; teniposide; edatrexate; daunomycin; aminopterin; capecitabine; ibandronate; CPT-11; topoisomerase inhibitor RFS2000; difluoromethylornithine (DMFO); retinoids, such as retinoic acid; and pharmaceutically acceptable salts, acids, and derivatives of any of the foregoing.

Chemotherapeutic agents also include: (i) antihormonal agents that act to modulate or inhibit the effects of hormones on tumors, such as antiestrogens and selective estrogen receptor modulators (SERMs), including, for example, tamoxifen (including tamoxifen citrate), raloxifene, droloxifene, iodoxyfene, 4-hydroxytamoxifen, trioxifene, keoxifene, LY117018, onapristone, and toremifene citrate; (ii) aromatase inhibitors that inhibit the enzyme aromatase that regulates estrogen production in the adrenal glands, such as, for example, 4(5)-imidazole, aminoglutethimide, megestrol acetate, and dapoxetine. (iii) antiandrogens, such as flutamide, nilutamide, bicalutamide, leuprolide, and goserelin; buserelin, tripterelin, medroxyprogesterone acetate, and dapoxetine. acetate), diethylstilbestrol, premarin, fluoxymesterone, all-trans retinoic acid, fenretinide, and troxacitabine (a 1,3-dioxolane nucleoside cytosine analog); (iv) protein kinase inhibitors; (v) lipid kinase inhibitors; (vi) antisense oligonucleotides, particularly those that inhibit the expression of genes in signal transduction pathways involved in abnormal cell proliferation, such as, for example, PKC-α, Ralf and H-Ras; (vii) ribozymes such as VEGF expression inhibitors (for example) and HER2 expression inhibitors; (viii) vaccines such as gene therapy vaccines, for example, and rIL-2; topoisomerase 1 inhibitors such as rmRH; and (ix) pharmaceutically acceptable salts, acids and derivatives of any of the above substances.

Chemotherapeutic agents also include antibodies such as alemtuzumab (Campath), bevacizumab (Genentech); cetuximab (Imclone); panitumumab (Amgen), rituximab (Genentech/Biogen Idec), pertuzumab (2C4, Genentech), trastuzumab (Genentech), tositumomab (Bexxar, Corixia), and the antibody-drug conjugate gemtuzumab ozogamicin (Wyeth). Other humanized monoclonal antibodies with therapeutic potential as chemotherapeutic agents for use in combination with the compounds of the invention include apolizumab, aselizumab, atlizumab, bapineuzumab, bivatuzumab mertansine, cantuzumab mertansine, cedelizumab, certolizumab pegol, and silizumab. pegol), cidfusituzumab, cidtuzumab, daclizumab, eculizumab, efalizumab, epratuzumab, erlizumab, felvizumab, fontolizumab, gemtuzumab ozogamicin, inotuzumabozogamicin, ipilimumab, labetuzumab, lintuzumab, matuzumab, mepolizumab, motavizumab, motovizumab, natalizumab, nimotuzumab, nolovizumab, numavizumab, ocreli zumab, omalizumab, palivizumab, pascolizumab, pecfusituzumab, pectuzumab, pexelizumab, ralivizumab, ranibizumab, reslivizumab, reslizumab, resyvizumab, rovelizumab, ruplizumab, sibrotuzumab, siplizumab, sontuzumab, tacatuzumab tetraxetan, tadocizumab, talizumab, tefibazumab, tocilizumab, toralizumab, tucotuzumab celmoleukin, tucusituzumab, umavizumab, urtoxazumab, ustekinumab, visilizumab, and anti-interleukin-12 (ABT-874/J695, Wyeth Research and Abbott Laboratories), which is an exclusively recombinant, human sequence, full-length IgG1 _lambda antibody genetically modified to recognize the interleukin-12p40 protein.

Chemotherapeutic agents also include "EGFR inhibitors," which refer to compounds that bind to or otherwise interact directly with EGFR and prevent or reduce its signaling activity, or are referred to as "EGFR antagonists." Examples of such agents include antibodies and small molecules that bind to EGFR. Examples of antibodies that bind to EGFR include MAb 579 (ATCC CRL HB 8506), MAb 455 (ATCC CRL HB8507), MAb 225 (ATCC CRL 8508), MAb 528 (ATCC CRL 8509) (see U.S. Patent No. 4,943,533, Mendelsohn et al.) and variants thereof, such as chimeric 225 (C225 or cetuximab;) and reconstructed human 225 (H225) (see WO 96/40210, Imclone Systems Inc.); fully human EGFR-targeting antibody IMC-11F8 (Imclone); antibodies that bind to type II mutant EGFR (U.S. Patent No. 5,212,290); humanized and chimeric antibodies that bind to EGFR as described in U.S. Patent No. 5,891,996; and human antibodies that bind to EGFR, such as ABX-EGF or Panitumumab (see WO98/50433, Abgenix/Amgen); EMD 55900 (Stragliotto et al. Eur. J. Cancer 32A:636-640 (1996)); humanized EGFR antibody EMD7200 (matuzumab) (EMD/Merck), which competes with both EGF and TGF-α for EGFR binding; human EGFR antibody, HuMax-EGFR (GenMab); fully human antibodies designated E1.1, E2.4, E2.5, E6.2, E6.4, E2.11, E6.3, and E7.6.3 and described in U.S. Pat. No. 6,235,883; MDX-447 (Medarex Inc); and mAb 806 or humanized mAb 806 (Johns et al., J. Biol. Chem. 279(29):30375-30384 (2004)). Anti-EGFR antibodies can be conjugated to cytotoxic agents to generate immunoconjugates (see, e.g., EP 659,439 A2, Merck Patent GmbH). EGFR antagonists include small molecules such as those described in U.S. Patent Nos. 5,616,582, 5,457,105, 5,475,001, 5,654,307, 5,679,683, 6,084,095, 6,265,410, 6,455,534, 6,521,620, 6,596,726, 6,713,484, 5,770,599, 6,140,3 32, 5,866,572, 6,399,602, 6,344,459, 6,602,863, 6,391,874, 6,344,455, 5,760,041, 6,002,008, and 5,747,498, and the following PCT publications: WO 98/14451, WO 98/50038, WO 99/09016, and WO 99/24037. Specific small molecule EGFR antagonists include OSI-774 (CP-358774, erlotinib, Genentech/OSI Pharmaceuticals); PD 183805 (CI 1033, 2-acrylamide, N-[4-[(3-chloro-4-fluorophenyl)amino]-7-[3-(4-morpholino)propoxy]-6-quinazolinyl]-, dihydrochloride, Pfizer Inc.); ZD1839, gefitinib, 4-(3'-chloro-4'-fluoroanilino)-7-methoxy-6-(3-morpholinopropoxy)quinazoline, AstraZeneca); ZM 105180 ((6-amino-4-(3-methylphenyl-amino)-quinazoline, Zeneca); BIBX-1382 (N8-(3-chloro-4-fluoro-phenyl)-N2-(1-methyl-piperidin-4-yl)-pyrimido[5,4-d]pyrimidine-2,8-diamine, Boehringer Ingelheim); PKI-166 ((R)-4-[4-[(1-phenylethyl)amino]-1H-pyrazolo[2,3-d]pyrimidin-6-yl]phenol); (R)-6-(4-hydroxyphenyl)-4-[(1-phenylethyl)amino]-7H-pyrazolo[2,3-d]pyrimidine); CL-387785 (N-[4-[(3-bromophenyl)amino]-6-quinazolinyl]-2-butynamide); EKB-569 (N-[4-[(3-chloro-4-fluorophenyl)amino]-3-cyano-7-ethoxy-6-quinolinyl]-4-(dimethylamino)-2-butenamide) (Wyeth); AG1478 (Pfizer); AG1571 (SU 5271; Pfizer); dual EGFR/HER2 tyrosine kinase inhibitors such as lapatinib (GSK572016 or N-[3-chloro-4-[(3-fluorophenyl)methoxy]phenyl]-6[5[[[2-methylsulfonyl)ethyl]amino]methyl]-2-furyl]-4-quinazolinamine).

Chemotherapeutic agents also include the "tyrosine kinase inhibitors" of the EGFR-targeted drugs described in the preceding paragraphs; small molecule HER2 tyrosine kinase inhibitors such as TAK165 available from Takeda; CP-724,714, oral selective inhibitors of ErbB2 receptor tyrosine kinase (Pfizer and OSI); dual HER inhibitors such as EKB-569 (available from Wyeth), which preferentially binds to EGFR but inhibits both HER2 and EGFR-overexpressing cells; lapatinib (GSK572016; available from Glaxo-SmithKline), an oral HER2 and EGFR tyrosine kinase inhibitor; PKI-166 (available from Novartis); pan-HER inhibitors such as canertinib (CI-1033; Pharmacia); Raf-1 inhibitors such as the antisense agent ISIS-5132 available from ISIS Pharmaceuticals, which inhibits Raf-1 signaling; non-HER-targeted TK inhibitors such as imatinib mesylate (available from Glaxo-SmithKline); SmithKline); multi-target tyrosine kinase inhibitors such as sunitinib (available from Pfizer); VEGF receptor tyrosine kinase inhibitors such as vatalanib (PTK787/ZK222584, available from Novartis/Schering AG); MAPK extracellular-regulated kinase I inhibitor CI-1040 (available from Pharmacia); quinazolines such as PD 153035, 4-(3-chloroanilino)quinazoline; pyridopyrimidines; pyrimidopyrimidines; pyrazolopyrimidines such as CGP 59326, CGP 60261, and CGP 62706; pyrazolopyrimidines, 4-(phenylamino)-7H-pyrazolo[2,3-d]pyrimidines; curcumin (diferuloylmethane, 4,5-bis(4-fluoroanilino)phthalimide); tyrphostines containing a nitrothiophene moiety; PD-0183805 (Warner-Lamber); antisense molecules (e.g., those that bind to HER-encoding nucleic acids); quinoxalines (U.S. Patent No. 5,804,396); tryphostins (U.S. Patent No. 5,804,396); ZD6474 (Astra Zeneca); PTK-787 (Novartis/Schering AG); the pan-HER inhibitor human CI-1033 (Pfizer); Affinitac (ISIS 3521; Isis/Lilly); imatinib mesylate PKI 166 (Novartis); GW2016 (Glaxo SmithKline); CI-1033 (Pfizer); EKB-569 (Wyeth); Semaxinib (Pfizer); ZD6474 (AstraZeneca); PTK-787 (Novartis/Schering AG); INC-1C11 (Imclone), sirolimus; or those described in any of the following patent publications: U.S. Patent No. 5,804,396; WO 1999/09016 (American Cyanamid); WO 1998/43960 (American Cyanamid); WO 1997/38983 (Warner Lambert); WO 1999/06378 (Warner Lambert); WO1999/06396 (Warner Lambert); WO 1996/30347 (Pfizer, Inc); WO 1996/33978 (Zeneca); WO1996/3397 (Zeneca) and WO 1996/33980 (Zeneca).

Chemotherapeutic agents also include dexamethasone, interferon, colchicine, metoprine, cyclosporine, amphotericin, metronidazole, alemtuzumab, alitretinoin, allopurinol, amifostine, arsenic trioxide, asparaginase, BCG live, bevacizumab, bexarotene, cladribine, clofarabine, darbepoetin alfa, denileukin, dexrazoxane, epoetin alfa, and dapoxetine. alfa), erlotinib, filgrastim, histrelin acetate, ibritumomab, interferon alfa-2a, interferon alfa-2b, lenalidomide, levamisole, mesna, methoxsalen, nandrolone, nelarabine, nofetumomab, oprelvekin, palifermin, pamidronate, pegademase, pegaspargase, pegfilgrastim, pemetrexed disodium, plicamycin, porfimer sodium sodium), quinacrine, rasburicase, sargramostim, temozolomide, VM-26, 6-TG, toremifene, tretinoin, ATRA, valrubicin, zoledronate and zoledronic acid and pharmaceutically acceptable salts thereof.

Chemotherapeutic agents also include hydrocortisone, hydrocortisone acetate, cortisone acetate, tixocortol pivalate, triamcinolone acetonide, triamcinolone alcohol, mometasone, amcinonide, budesonide, desonide, fluocinonide, fluocinolone acetonide, and fluocinolone acetonide. acetonide), betamethasone, betamethasone sodium phosphate, dexamethasone, dexamethasone sodium phosphate, fluocortolone, hydrocortisone-17-butyrate, hydrocortisone-17-valerate, aclometasone dipropionate, betamethasone valerate, betamethasone dipropionate, prednicarbate, clobetasone-17-butyrate, clobetasone-17-propionate, fluocortolone hexanoate, fluocortolone pivalate, and fluprednidene acetate; immunoselective anti-inflammatory peptides (ImSAIDs) such as phenylalanine-glutamine-glycine (FEG) and its D-isomer form (FEG) (IMULAN BioTherapeutics, LLC; antirheumatic drugs such as azathioprine, ciclosporin (cyclosporine A), D-penicillamine, gold salts, hydroxychloroquine, leflunomide, minocycline, sulfasalazine, tumor necrosis factor alpha (TNFα) blockers such as etanercept (Enbrel), infliximab (Remicade), adalimumab (Humira), certolizumab pegol) (Cimzia), golimumab (Simponi), interleukin 1 (IL-1) blockers such as anakinra (Kineret), T cell costimulation blockers such as abatacept (Orencia), interleukin 6 (IL-6) blockers such as tocilizumab interleukin 13 (IL-13) blockers such as lebrikizumab; interferon alpha (IFN) blockers such as rontalizumab; β7 integrin blockers such as rhuMAb Beta7; IgE pathway blockers such as anti-M1 primer; secreted homotrimeric LTa3 and membrane-bound heterotrimeric LTa1/β2 blockers such as anti-lymphotoxin alpha (LTa); radioactive isotopes (e.g., At ²¹¹ , I ¹³¹ , I ¹²⁵ , Y ⁹⁰ , Re ¹⁸⁶ , Re ¹⁸⁸ , Sm ¹⁵³ , Bi ²¹² , P ³² , Pb ²¹² , and radioisotopes of Lu); various investigational agents such as thioplatin, PS-341, phenylbutyrate, ET-18-OCH ₃ , or farnesyltransferase inhibitors (L-739749, L-744832); polyphenols such as quercetin, resveratrol, piceatannol, epigallocatechine gallate, theaflavin, flavanols, procyanidins, betulinic acid, acid and its derivatives; autophagy inhibitors such as chloroquine; delta-9-tetrahydrocannabinol (dronabinol); beta-lapachone; lapachol; colchicine; betulinic acid; acetylcamptothecin, scopolectin, and 9-aminocamptothecin; podophyllotoxin; tegafur; bexarotene; bisphosphonates such as clodronate (e.g., clodronate or etidronate); etidronate NE-58095, zoledronic acid/zoledronic acid alendronate pamidronate tiludronate or risedronate and epidermal growth factor receptor (EGF-R); vaccines such as vaccines; perifosine, COX-2 inhibitors (e.g., celecoxib or etoricoxib), proteasome inhibitors (e.g., PS341); CCI-779; tipifarnib (R11577); orafenib, ABT510; Bcl-2 inhibitors such as oblimersen farnesyltransferase inhibitors such as lonafarnib (SCH 6636, SARASAR ^™ ); and pharmaceutically acceptable salts, acids, or derivatives of any of the foregoing agents; and combinations of two or more of the foregoing agents, such as CHOP, an abbreviation for cyclophosphamide, doxorubicin, vincristine, and prednisolone combination therapy; and FOLFOX, an abbreviation for oxaliplatin (ELOXATIN ^™ ) in combination with 5-FU and folinic acid.

Chemotherapeutic agents also include nonsteroidal anti-inflammatory drugs (NSAIDs), which have analgesic, antipyretic, and anti-inflammatory effects. NSAIDs include non-selective inhibitors of the enzyme cyclooxygenase. Specific examples of NSAIDs include aspirin; propionic acid derivatives such as ibuprofen, fenoprofen, ketoprofen, flurbiprofen, oxaprozin, and naproxen; acetic acid derivatives such as indomethacin, sulindac, etodolac, and diclofenac; enolic acid derivatives such as piroxicam, meloxicam, tenoxicam, droxicam, lornoxicam, and isoxicam; fenamic acid derivatives such as mefenamic acid, meclofenamic acid, flufenamic acid, and fenamic acid. NSAIDs are used for symptomatic relief of conditions such as rheumatoid arthritis, osteoarthritis, inflammatory arthritis, ankylosing spondylitis, psoriatic arthritis, Reiter's syndrome, acute gout, dysmenorrhea, metastatic bone pain, headache and migraine, postoperative pain, mild to moderate pain due to inflammation and tissue damage, fever, intestinal obstruction and renal colic.

In certain embodiments, chemotherapeutic agents include but are not limited to doxorubicin, dexamethasone, vincristine, cyclophosphamide, fluorouracil, topotecan, interferon, platinum derivatives, taxanes (e.g., paclitaxel, docetaxel), vinca alkaloids (e.g., vinblastine), anthracyclines (e.g., doxorubicin), epipodophyllotoxins (e.g., etoposide), cisplatin, mTOR inhibitors (e.g., rapamycin), methotrexate, dactinomycin D, dolastatin 10, colchicine, trimetrexate, metropine, cyclosporine, daunorubicin, teniposide, amphotericin, alkylating agents (e.g., chlorambucil), 5-fluorouracil, campthothecin, cisplatin, metronidazole, and imatinib mesylate, etc. In other embodiments, the compounds of the present invention are administered in combination with a biologic such as bevacizumab or panitumumab.

In certain embodiments, a compound of the present invention or a pharmaceutically acceptable composition thereof is administered in combination with an antiproliferative or chemotherapeutic agent selected from any one or more of the following: abarelix, aldesleukin, alemtuzumab, acitretin, allopurinol, hexamethylmelamine, amifostine, anastrozole, arsenic trioxide, asparaginase, azacitidine, live bacille Calmette-Guérin, bevacizumab, fluorouracil, bexarotene, bleomycin, bortezomib, busulfan, kalutestosterone, capecitabine, camptothecin, carboplatin, carmustine, Cetuximab, chlorambucil, cladribine, clofabamine, cyclophosphamide, cytarabine, dactinomycin, darbepoetin alfa, aclarubicin, denileukin, dexrazoxane, docetaxel, doxorubicin (neutral), doxorubicin hydrochloride, drostanolone propionate, epirubicin, epoetin alfa, erlotinib, estramustine, etoposide phosphate, etoposide, exemestane, filgrastim, floxuridine, fludarabine, fulvestrant, gefitinib, gemcitabine, gemtuzumab, goserelin acetate, histrelin acetate, hydroxyurea, ibritumomab tiuxetan Anti-inflammatory drugs, idarubicin, ifosfamide, imatinib mesylate, interferon alfa-2a, interferon alfa-2b, irinotecan, lenalidomide, letrozole, leucovorin, leuprorelin acetate, levamisole, lomustine, megestrol acetate, melphalan, mercaptopurine, 6-MP, mesna, methotrexate, methoxsalen, mitomycin C, mitotane, mitoxantrone, nandrolone, nelarabine, nofel tuzumab, oprelvekin, oxaliplatin, paclitaxel, palifermin, pamidronate, pegaspargase, pegfilgrastim , pemetrexed disodium, pentostatin, pipobroman, pramycin, porfimer sodium, procarbazine, quinacrine, rasburicase, rituximab, sarcoma, sorafenib, streptozocin, sunitinib maleate, talc, tamoxifen, temozolomide, teniposide, VM-26, testolactone, thioguanine, 6-TG, thiotepa, topotecan, toremifene, tositumomab, trastuzumab, tretinoin, ATRA, uracil mustard, valrubicin, vinblastine, vincristine, vinorelbine, zoledronic acid, or zoledronic acid.

Chemotherapeutic agents also include treatments for Alzheimer's disease, such as donepezil hydrochloride and rivastigmine; treatments for Parkinson's disease, such as L-DOPA/carbidopa, entacapone, ropinrole, pramipexole, bromocriptine, pergolide, trihexephendyl, and amantadine; agents used to treat multiple sclerosis (MS), such as beta interferon (e.g., levofloxacin and tadalafil), glatiramer acetate, and mitoxantrone; treatments for asthma, such as albuterol and montelukast sodium. sodium); agents used to treat schizophrenia, such as Zyprexa, risperdal, Seroquel, and haloperidol; anti-inflammatory agents, such as corticosteroids, TNF blockers, IL-1RA, azathioprine, cyclophosphamide, and sulfasalazine; immunomodulators and immunosuppressants, such as cyclosporine, tacrolimus, rapamycin, mycophenolate mofetil, and tacrolimus. neurotrophic factors such as acetylcholinesterase inhibitors, MAO inhibitors, interferons, anticonvulsants, ion channel blockers, riluzole, and anti-Parkinson's disease agents; agents for treating cardiovascular disease such as beta-blockers, ACE inhibitors, diuretics, nitrates, calcium channel blockers, and statins; agents for treating liver disease such as corticosteroids, cholestyramine, interferon, and antiviral agents; agents for treating blood disorders such as corticosteroids, anti-leukemia agents, and growth factors; and agents for treating immunodeficiency disorders such as gamma globulin.

Additionally, chemotherapeutic agents include pharmaceutically acceptable salts, acids, or derivatives of any chemotherapeutic agent described herein, and combinations of two or more thereof.

For the treatment of inflammatory or autoimmune diseases, it can be used with methotrexate, tofacitinib, 6-mercaptopurine, azathioprine, sulfasalazine, mesalazine, olsalazine, chloroquine/hydroxychloroquine, penicillamine, aurothiomalate (intramuscular and oral), azathioprine, colchicine, corticosteroids (oral, inhaled and local injection), beta-2 adrenergic receptor agonists (salbutamol, terbutaline, salmeteral), xanthines (theophylline, aminophylline), cromoglycate, nedocromil, ketotifen, ipratropium and oxitropium, cyclosporine, FK506, rapamycin, mycophenolate mofetil, leflunomide, NSAIDs (e.g., ibuprofen), corticosteroids (e.g., prednisolone), phosphodiesterase inhibitors, adenosine agonists, antithrombotic agents, complement inhibitors, adrenergic agents, agents that interfere with signaling through proinflammatory cytokines such as TNF or IL-1 (e.g., NIK, IKK, p38 or MAP kinase inhibitors), IL-1 converting enzyme inhibitors, T-cell signaling inhibitors (e.g., kinase inhibitors), metalloproteinase inhibitors, sulfasalazine, 6-mercaptopurine, angiotensin converting enzyme inhibitors, soluble cytokine receptors (e.g., soluble p55 or p75 TNF receptors and derivatives p75TNFRigG (etanercept) and p55TNFRigG (Lenercept), siL-1RI, siL-1RII, siL-6R), anti-inflammatory cytokines (e.g., IL-4, IL-1 0, IL-11, IL-13, and TGF-β), celecoxib, folic acid, hydroxychloroquine sulfate, rofecoxib, etanercept, infliximab, adalimumab, certolizumab pegol, tocilizumab, abatacept, naproxen, valdecoxib, sulfasalazine, methylprednisolone, meloxicam, methylprednisolone acetate, sodium aurothiomalate, aspirin, triamcinolone acetonide, propoxyphene napsylate/acetaminophen (apap), folic acid (folate), nabumetone, diclofenac, piroxicam, etodolac, diclofenac sodium, oxaprozin, oxycodone HCl, hydrocodone bitartrate bitartrate/acetaminophen, diclofenac sodium/misoprostol, fentanyl, anakinra, tramadol HCl, salsalate, sulindac, cyanocobalamin/fa/pyridoxine, acetaminophen, alendronate sodium, prednisolone, cortisone, betamethasone, morphine sulfate, lidocaine hydrochloride, indomethacin, glucosamine sulfate In some embodiments, the present invention includes but is not limited to: (i) sulf/chondroitin, amitriptyline HCl, sulfadiazine, oxycodone HCl/acetaminophen, olopatadine HCl, misoprostol, naproxen sodium, omeprazole, cyclophosphamide, rituximab, IL-1 TRAP, MRA, CTLA4-IG, IL-18BP, anti-IL-12, anti-IL1S, BIRB-796, SCIO-469, VX-702, AMG-548, VX-740, roflumilast, IC-485, CDC-801, S1P1 agonists (such as FTY720), PKC family inhibitors (e.g., ruboxistaurin or AEB-071), or mesopram. In certain embodiments, the compound of formula (I) or a pharmaceutically acceptable salt thereof can be co-administered with methotrexate or leflunomide. In moderate or severe rheumatoid arthritis cases, the compound of formula (I) or a pharmaceutically acceptable salt thereof can be co-administered with the above-mentioned cyclosporine and anti-TNF antibodies. The compound of formula (I) or a pharmaceutically acceptable salt thereof can also be co-administered with the following agents: budesonide; epidermal growth factor; corticosteroids; cyclosporin, sulfasalazine; para-aminosalicylate; 6-mercaptopurine; azathioprine; metronidazole; lipoxygenase inhibitors; mesalamine; olsalazine; balsalazide; antioxidants; thromboxane inhibitors; IL-1 receptor antagonists; anti-IL-1 monoclonal antibodies; anti-IL-6 monoclonal antibodies; growth factors; elastase inhibitors; pyridyl-imidazole compounds; anti-human cytokines or growth factors (e.g., TNF, LT, IL-1, IL-2, IL-6, I Antibodies to IL-7, IL-8, IL-12, IL-15, IL-16, IL-23, EMAP-II, GM-CSF, FGF, and PDGF, or their antagonists; cell surface molecules (e.g., CD2, CD3, CD4, CD8, CD25, CD28, CD30, CD40, CD45, CD69, or CD90, or their ligands); methotrexate; cyclosporine; FK506; rapamycin; mycophenolate mofetil; leflunomide; NSAIDs (e.g., ibuprofen); corticosteroids (e.g., prednisolone); phosphodiesterase inhibitors; adenosine agonists; antithrombotic agents; complement inhibitors; adrenergic agents; agents that interfere with signaling by proinflammatory cytokines, such as TNF 5 or IL-1 (e.g., NIK, IKK or MAP kinase inhibitors); IL-1 converting enzyme inhibitors; TNF converting enzyme inhibitors; T-cell signaling inhibitors such as kinase inhibitors; metalloproteinase inhibitors; sulfasalazine; azathioprine; 6-mercaptopurine; angiotensin-converting enzyme inhibitors; soluble cytokine receptors (e.g., soluble p55 or p75 TNF receptor, siL-1RI, siL-1RII, siL-6R) and anti-inflammatory cytokines (e.g., IL-4, IL-1 β, IL-11, IL-13 or TGF-β).

For the treatment of Crohn's disease, a compound of formula (I) or a pharmaceutically acceptable salt thereof may be co-administered with a TNF antagonist (e.g., an anti-TNF antibody), D2E7 (adalimumab), CA2 (infliximab), CDP 571, a TNFR-Ig construct, (p75TNFRigG (etanercept)), a p55TNFRigG (LENERCEPT ^™ ) inhibitor, or a PDE4 inhibitor.

For the treatment of inflammatory bowel disease, a compound of formula (I) or a pharmaceutically acceptable salt thereof can be co-administered with the following agents: corticosteroids (e.g., budesonide or dexamethasone); sulfasalazine, 5-aminosalicylic acid; olsalazine; agents that interfere with the synthesis or action of proinflammatory cytokines such as IL-1 (e.g., IL-1 converting enzyme inhibitors or IL-1ra); T cell signaling inhibitors (e.g., tyrosine kinase inhibitors); 6-mercaptopurine; IL-11; mesalazine; prednisone; azathioprine; mercaptopurine; infliximab; methylprednisolone sodium succinate; diphenoxylate/atropin sulfate; loperamide hydrochloride; methotrexate; omeprazole; folic acid; ciprofloxacin/dextrose-water; hydrocodone bitartrate/acetaminophen; tetracycline hydrochloride hydrochloride; fluocinolone acetonide; metronidazole; thimerosal/boric acid; cholestyramine/sucrose; ciprofloxacin hydrochloride; scopolamine sulfate; meperidine hydrochloride; midazolam hydrochloride; oxycodone hydrochloride/acetaminophen; promethazine hydrochloride; sodium phosphate; sulfamethoxazole/trimethoprim; celecoxib; polycarbophil; propoxyphene napsylate; hydrocortisone; multivitamin; balsalazide disodium; codeine phosphate/acetaminophen; colesevelam HCl; cyanocobalamin; folic acid; levofloxacin; methylprednisolone; natalizumab or interferon-gamma.

For the treatment of multiple sclerosis, the compound of formula (I) or a pharmaceutically acceptable salt thereof can be co-administered with the following agents: corticosteroids; prednisolone; methylprednisolone; azathioprine; cyclophosphamide; cyclosporine; methotrexate; 4-aminopyridine; tizanidine; interferon-1a (Biogen); interferon-1b (Chiron/Berlex); interferon-n3 (Interferon Sciences/Fujimoto), interferon- (Alfa Wassermann/J&J), interferon 1A-IF (Serono/Inhale Therapeutics), pegylated interferon 2b (Enzon/Schering-Plough), copolymer 1 (Cop-1; Teva Pharmaceuticals Industries, Inc.); hyperbaric oxygen; intravenous immunoglobulin; cladribine; antibodies to other human cytokines or growth factors and their receptors (e.g., TNF, LT, IL-1, IL-2, IL-6, IL-7, IL-8, IL-12, IL-23, IL-15, IL-16, EMAP-II, GM-CSF, FGF, or PDGF) or their antagonists

For the treatment of AIDS, a compound of formula (I) or a pharmaceutically acceptable salt thereof can be co-administered with antibodies directed against cell surface molecules such as CD2, CD3, CD4, CD8, CD19, CD20, CD25, CD28, CD30, CD40, CD45, CD69, CD80, CD86, CD90 or their ligands. A compound of formula (I) or a pharmaceutically acceptable salt thereof can also be co-administered with methotrexate, cyclosporine, FK506, rapamycin, mycophenolate mofetil, leflunomide, an S1P1 agonist, an NSAID (e.g., ibuprofen), a corticosteroid (e.g., prednisolone), a phosphodiesterase inhibitor, an adenosine agonist, an antithrombotic agent, a complement inhibitor, an adrenergic agent, an inhibitor of inflammatory bowel disease (IBD) or an anti-inflammatory drug (e.g., TNF-α or IL-1), or an inhibitor of TNF-α or IL-2 (e.g., NIK, IKK, p38 or MAB). [00135] The present invention also includes agents that interfere with signaling (e.g., IL-1 kinase inhibitors), IL-1 converting enzyme inhibitors, TACE inhibitors, T-cell signaling inhibitors (e.g., kinase inhibitors), metalloproteinase inhibitors, sulfasalazine, azathioprine, 6-mercaptopurine, angiotensin converting enzyme inhibitors, soluble cytokine receptors (e.g., soluble p55 or p75 TNF receptors, siL-1RI, siL-1RII, or siL-6R), or anti-inflammatory cytokines (e.g., IL-4, IL-1β, IL-13, or TGF-β).

The compound of formula (I) or a pharmaceutically acceptable salt thereof can also be co-administered with agents such as alemtuzumab, dronabinol, daclizumab, mitoxantrone, xaliproden hydrochloride, fampridine, glatiramer acetate, natalizumab, sinnabidol, immunokine NNS03, ABR-215062, AnergiX.MS, chemokine receptor antagonists, BBR-2778, calagualine, CPI-1189, LEM (liposomally encapsulated mitoxantrone), THC.CBD (cannabinoid agonist), MBP-8298, mesopram (PDE4 inhibitor), MNA-715, anti-IL-6 receptor antibody, neurovax, pirfenidone allotrap 1258 (RDP-1258), sTNF-R1, talampanel, teriflunomide, TGF-β2, tiplimotide, a VLA-4 antagonist (e.g., TR-14035, VLA4 Ultrahaler, or Antegran-ELAN/Biogen), an interferon gamma antagonist, or an IL-4 agonist.

For the treatment of ankylosing spondylitis, the compound of formula (I) or a pharmaceutically acceptable salt thereof can be co-administered with the following agents: ibuprofen, diclofenac, misoprostol, naproxen, meloxicam, indomethacin, diclofenac, celecoxib, rofecoxib, sulfasalazine, methotrexate, azathioprine, minocycline, prednisone, anti-TNF antibodies, D2E7CA2 (infliximab), CDP 571, TNFR-Ig constructs, (p75TNFRigG or p55TNFRigG

For the treatment of asthma, the compound of formula (I) or a pharmaceutically acceptable salt thereof can be co-administered with the following agents: salbutamol, salmeterol/fluticasone, montelukast sodium, fluticasone propionate, budesonide, prednisone, salmeterol xinafoate, levalbuterol HCl, salbutamol sulfate/ipratropium, prednisolone sodium phosphate, triamcinolone acetonide, beclomethasone dipropionate, ipratropium bromide, azithromycin, pirbuterol acetate, prednisolone, theophylline anhydrous, methylprednisolone sodium succinate, clarithromycin, zafirlukast, formoterol fumarate, influenza virus vaccine, amoxicillin trihydrate, trihydrate), flunisolide, sodium cromoglycate, fexofenadine hydrochloride, flunisolide/menthol, amoxicillin/clavulanate, levofloxacin, guaifenesin, dexamethasone sodium phosphate, moxifloxacin HCl, doxycycline hyclate, guaifenesin/d-methorphan, p-ephedrine/cod/-chlorphenir, gatifloxacin, cetirizine hydrochloride, mometasone furoate furoate, salmeterol xinafoate, benzonatate, cephalexin, PE/hydrocodone/chlorpheniramine, cetirizine hydrochloride/pseudoephedrine, phenylephrine/COD/promethazine, codeine/promethazine, cefprozil, dexamethasone, guaifenesin/pseudoephedrine, chlorpheniramine/hydrocodone, nedocromil sodium, terbutaline sulfate, epinephrine, methylprednisolone, anti-IL-13 antibodies, or metaproterenol sulfate.

For the treatment of COPD, the compound of formula (I) or a pharmaceutically acceptable salt thereof can be co-administered with the following agents: salbutamol sulfate/ipratropium, ipratropium bromide, salmeterol/fluticasone, salbutamol, salmeterol xinafoate, fluticasone propionate, prednisone, theophylline anhydrous, methylprednisolone sodium succinate, montelukast sodium, budesonide, formoterol fumarate, triamcinolone acetonide, levofloxacin, guaifenesin, azithromycin, beclomethasone dipropionate, levosalbutamol hydrochloride, flunisolide, ceftriaxone sodium sodium), amoxicillin trihydrate, gatifloxacin, zafirlukast, amoxicillin/clavulanate, flunisolide/menthol, chlorpheniramine/hydrocodone, metaproterenol sulfate, methylprednisolone, mometasone furoate, p-ephedrine/cod/chlorpheniramine, pirbuterol acetate, p-ephedrine/loratadine, terbutaline sulfate, tiotropium bromide, (R,R)-formoterol, TgAAT, cilomilast, or roflumilast.

For the treatment of psoriasis, the compound of formula (I) or a pharmaceutically acceptable salt thereof can be co-administered with the following agents: calcipotriene, clobetasol propionate, triamcinolone acetonide, halobetasol propionate, tazarotene, methotrexate, fluocinolone acetonide, betamethasone dipropionate augmented, fluocinolone acetonide, acitretin, tar shampoo, shampoo), betamethasone valerate, mometasone furoate, ketoconazole, pramoxine/fluocinolone, hydrocortisone valerate, flurandrenolide, urea, betamethasone, clobetasol propionate/emoll, fluticasone propionate, azithromycin, hydrocortisone, moisturizing formula, folic acid, desonide, pimecrolimus, coal tar, diflorasone diacetate, etanercept, folic acid, lactic acid, methoxsalen, bismuth subgallate subgal/znox/resor, methylprednisolone acetate, prednisone, sunscreen, halcinonide, salicylic acid, anthralin, clocortolone pivalate, coal extract, coal tar/salicylic acid, coal tar/salicylic acid/sulfur, desoximetasone, diazepam, emollient, fluocinolone acetonide/emollient, mineral oil/castor oil/nalact, mineral oil/peanut oil, petroleum/isopropyl myristate, psoralen, salicylic acid, soap/tribromsalan, thimerosal/boric acid, celecoxib, infliximab, cyclosporine, alefacept, efalizumab, tacrolimus, pimecrolimus, PUVA, UVB, sulfasalazine, ABT-874, or ustekinamab.

For the treatment of psoriatic arthritis, the compound of formula (I) or a pharmaceutically acceptable salt thereof can be co-administered with the following agents: methotrexate, etanercept, rofecoxib, celecoxib, folic acid, sulfasalazine, naproxen, leflunomide, methylprednisolone acetate, indomethacin, hydroxychloroquine sulfate, prednisone, sulindac, betamethasone dipropionate boosted, infliximab, methotrexate, folic acid, triamcinolone acetonide, diclofenac, dimethyl sulfoxide, piroxicam, diclofenac sodium, ketoprofen, meloxicam, methylprednisolone, nabumetone, tolmetin sodium, calcipotriene, cyclosporine, diclofenac sodium/misoprostol, fluocinolone acetonide, glucosamine sulfate, sodium aurothiomalate, hydrocodone bitartrate/acetaminophen, ibuprofen, risedronate sodium sodium), sulfadiazine, thioguanine, valdecoxib, alefacept, D2E7 (adalimumab), or efalizumab.

For the treatment of lupus, a compound of formula (I) or a pharmaceutically acceptable salt thereof can be co-administered with an NSAID (e.g., diclofenac, naproxen, ibuprofen, piroxicam, or indomethacin); a COX2 inhibitor (e.g., celecoxib, rofecoxib, or valdecoxib); an antimalarial (e.g., hydroxychloroquine); a steroid (e.g., prednisone, prednisolone, budesonide, or dexamethasone); a cytotoxic agent (e.g., azathioprine, cyclophosphamide, mycophenolate mofetil, or methotrexate); an inhibitor of PDE4 or a purine synthesis inhibitor (e.g., sulfasalazine, 5-aminosalicylic acid, olsalazine, an agent that interferes with the synthesis, production, or action of a proinflammatory cytokine (e.g., IL-1)); or a caspase inhibitor (e.g., an IL-1 converting enzyme inhibitor or IL-1ra).

The compound of formula (I) or a pharmaceutically acceptable salt thereof can also be co-administered with the following agents: T cell signaling inhibitors (such as tyrosine kinase inhibitors), or molecules targeting T cell activation (such as CTLA-4-IgG, anti-B7 family antibodies or anti-PD-1 family antibodies).

The compound of formula (I) or a pharmaceutically acceptable salt thereof can also be co-administered with the following agents: IL-11 antibody, anti-cytokine antibody (such as fonotolizumab (anti-IFNg antibody)) or anti-receptor antibody (such as anti-IL-6 receptor antibody or antibodies against B-cell surface molecules).

A compound of formula (I), or a pharmaceutically acceptable salt thereof, may also be co-administered with LJP 394 (abetimus), agents that deplete or inactivate B-cells, such as rituximab (anti-CD20 antibody) or lymphostat-B (anti-BlyS antibody), TNF antagonists, such as anti-TNF antibodies, D2E7 (adalimumab), CA2 (infliximab), CDP 571, TNFR-Ig constructs, (p75TNFRigG (etanercept) or p55TNFRigG (LENERCEPT ^™ ).

The compound of formula (I) or a pharmaceutically acceptable salt thereof can also be co-administered with one or more agents for the prevention or treatment of AIDS: HIV reverse transcriptase inhibitors, HIV protease inhibitors, immunomodulators or other retroviral drugs. Examples of reverse transcriptase inhibitors include, but are not limited to, abacavir, adefovir, didanosine, dipivoxil delavirdine, efavirenz, emtricitabine, lamivudine, nevirapine, rilpivirine, stavudine, tenofovir, zalcitabine and zidovudine. Examples of protease inhibitors include, but are not limited to, amprenavir, atazanavir, darunavir, indinavir, fosamprenavir, lopinavir, nelfinavir, ritonavir, saquinavir, and tipranavir. Examples of other retroviral drugs include, but are not limited to, elvitegravir, enfuvirtide, maraviroc, and raltegravir.

For the treatment of type II diabetes, hepatic steatosis, insulin resistance, metabolic syndrome or related diseases, the compound of formula (I) or a pharmaceutically acceptable salt thereof can be co-administered with the following agents: insulin or insulin modified to improve the duration of action in the body; agents that stimulate insulin secretion, such as acetohexamide, chlorpropamide, glyburide, glimepiride, glipizide, glicazide, glycopyramide, gliquidone, rapaglinide, nateglinide, tolazamide or tolbutamide; as a glucagon-like peptide agonist agents that inhibit dipeptidyl peptidase IV, such as vildagliptin, sitagliptin, saxagliptin, linagliptin, alloglitin, or septagliptin; agents that bind to peroxisome proliferator-activated receptor gamma, such as rosiglitazone or pioglitazone; agents that reduce insulin resistance, such as metformin; or agents that reduce glucose absorption in the small intestine, such as acarbose, miglitol, or voglibose.

For the treatment of acute or chronic kidney disease, the compound of formula (I) or a pharmaceutically acceptable salt thereof can be co-administered with dopamine, a diuretic (e.g., furosemide), bumetanide, a thiazide, mannitol, calcium gluconate, sodium bicarbonate, albuterol, paricalcitol, doxercalciferol, cinacalcet, or bardoxalonemethyl.

The amount of the compound of formula (I) or its salt and the additional medicament (in those compositions comprising additional therapeutic agents as described above) that can be combined with carrier materials to produce a single dosage form will depend on the host being treated and the specific mode of administration. In certain embodiments, the compositions of the present invention are formulated so that a dosage of 0.01-100 mg/kg body weight/day of the present invention can be administered.

The additional therapeutic agent and the compound of formula (I) may act synergistically. Thus, the amount of the additional therapeutic agent in such compositions may be less than that required in a monotherapy using only that therapeutic agent, or may result in fewer side effects in the patient when a lower dose is used. In certain embodiments, a dose of 0.01-1,000 μg/kg body weight/day of the additional therapeutic agent may be administered in such compositions.

Provided herein are methods for prolonging the duration of response to a cytotoxic agent in an individual having cancer, comprising administering to the individual (a) an effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof and (b) an effective amount of a cytotoxic agent.

In certain embodiments of any of the methods, the cytotoxic agent is a targeted therapy. In certain embodiments, the targeted therapy is one or more of a GFR antagonist, a RAF inhibitor, and/or a PI3K inhibitor.

In certain embodiments of either method, the targeted therapy is an EGFR antagonist. In certain embodiments of either method, the EGFR antagonist is N-(3-ethynylphenyl)-6,7-bis(2-methoxyethoxy)-4-quinazolinamine and/or a pharmaceutically acceptable salt thereof. In certain embodiments, the EGFR antagonist is N-(3-ethynylphenyl)-6,7-bis(2-methoxyethoxy)-4-quinazolinamine. In certain embodiments, the EGFR antagonist is N-(4-(3-fluorobenzyloxy)-3-chlorophenyl)-6-(5-((2-(methylsulfonyl)ethylamino)methyl)furan-2-yl)quinazolin-4-amine, di-4-methylbenzenesulfonate or a pharmaceutically acceptable salt thereof (e.g., lapatinib).

In certain embodiments of either method, the targeted therapy is a RAF inhibitor. In certain embodiments, the RAF inhibitor is a BRAF inhibitor. In certain embodiments, the RAF inhibitor is a CRAF inhibitor. In certain embodiments, the BRAF inhibitor is vemurafenib. In certain embodiments, the RAF inhibitor is 3-(2-cyanopropyl-2-yl)-N-(4-methyl-3-(3-methyl-4-oxo-3,4-dihydroquinazolin-6-ylamino)phenyl)benzamide or a pharmaceutically acceptable salt thereof (e.g., AZ628 (CAS#878739-06-1)).

In certain embodiments of any of the methods, the targeted therapy is a PI3K inhibitor.

In certain embodiments of any of the methods, the cytotoxic agent is chemotherapy. In certain embodiments of any of the methods, the chemotherapy is a taxane. In certain embodiments, the taxane is paclitaxel. In certain embodiments, the taxane is docetaxel.

In certain embodiments of any of the methods, the cytotoxic agent is a platinum agent. In certain embodiments, the platinum agent is carboplatin. In certain embodiments, the platinum agent is cisplatin. In certain embodiments of any of the methods, the cytotoxic agent is a taxane and a platinum agent. In certain embodiments, the taxane is paclitaxel. In certain embodiments, the taxane is docetaxel. In certain embodiments, the platinum agent is carboplatin. In certain embodiments, the platinum agent is cisplatin.

In certain embodiments of either method, the cytotoxic agent is a vinca alkaloid. In certain embodiments, the vinca alkaloid is vinorelbine. In certain embodiments of either method, the chemotherapy is a nucleoside analog. In certain embodiments, the nucleoside analog is gemcitabine.

In certain embodiments of any of the methods, the cytotoxic agent is radiation therapy.

In certain embodiments of either method, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered concomitantly with a cytotoxic agent (e.g., targeted therapy, chemotherapy, and/or radiotherapy). In certain embodiments, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered before and/or concurrently with a cytotoxic agent (e.g., targeted therapy, chemotherapy, and/or radiotherapy).

The invention will now be illustrated by the following non-limiting examples.

Example

Experimental operation of intermediate A

Step 1:

6-Methyl-6,7-dihydropyrido[2,3-d]pyridazine-5,8-dione

A mixture of dimethyl pyridine-2,3-dicarboxylate (100.0 g, 512.37 mmol) in methylhydrazine (40% aqueous solution, 1000 mL) was heated under reflux for 15 h, at which point LCMS indicated that the reaction was complete. The mixture was concentrated under reduced pressure to give a mixture of the title compound and 7-methyl-6,7-dihydropyrido[2,3-d]pyridazine-5,8-dione (90.0 g, 99% yield) as a yellow solid. The crude material was used in the next step without further purification. LCMS M/Z (M+H) 178. (Ratio of desired product: by-product = 4:3)

Step 2:

8-Chloro-6-methylpyrido[2,3-d]pyridazin-5(6H)-one

A mixture of 6-methyl-6,7-dihydropyrido[2,3-d]pyridazine-5,8-dione and 7-methyl-6,7-dihydropyrido[2,3-d]pyridazine-5,8-dione (90.0 g, 508.02 mmol) in POCl ₃ (900 mL) was heated at reflux for 15 h, at which point LCMS indicated that the reaction was complete. The mixture was concentrated under reduced pressure. The residue was slowly quenched by adding a saturated aqueous solution of NaHCO ₃ (1000 mL), followed by extraction with ethyl acetate (3x1000 mL). The combined organic extracts were dried over Na ₂ SO ₄ and concentrated under reduced pressure. The residue was purified by silica gel chromatography (petroleum ether:ethyl acetate=1:1) to give the title compound (12.5 g, 13% yield) as a light red solid. ¹ H NMR (400MHz, DMSO-d6) δ9.20 (d, J = 3.6 Hz, 1H), 8.65 (d, J = 7.2 Hz, 1H), 7.96 (dd, J = 4.8, 6.4 Hz 1H), 3.72 (s, 3H). LCMS M/Z(M+H)196.

Experimental operation of intermediates B and C

Step 1:

6-methoxy-2-methyl-2,3-dihydrophthalazine-1,4-dione, and

7-Methoxy-2-methyl-2,3-dihydrophthalazine-1,4-dione

A mixture of 5-methoxyisobenzofuran-1,3-dione (4.0 g, 22.45 mmol) and methylhydrazine (40% aqueous solution, 7.76 g, 67.36 mmol) in ethanol (80 mL) was heated at reflux for 15 h, at which point LCMS indicated the reaction was complete. The mixture was concentrated under reduced pressure to give the title compound (3.60 g, 78% yield) (2:1 ratio of regioisomers) as a yellow solid. The crude material was used in the next step without further purification. LCMS M/Z (M+H) 207.

Step 2:

4-chloro-6-methoxy-2-methylphthalazin-1(2H)-one, and

4-Chloro-7-methoxy-2-methylphthalazin-1(2H)-one

A mixture of 6- and 7-methoxy-2-methyl-2,3-dihydrophthalazine-1,4-dione (2.7 g, 13.09 mmol) in _POCl₃ (50 mL) was heated at reflux for 15 h, at which point LCMS indicated the reaction was complete. The mixture was concentrated under reduced pressure, and the residue was quenched by adding a saturated aqueous solution of _NaHCO₃ (100 mL), followed by extraction with ethyl acetate ( ₃ x 100 mL). The combined organic extracts were dried over _Na₂SO₄ and concentrated under reduced pressure. The residue was purified by silica gel chromatography (petroleum ether:ethyl acetate = 5:1) to give the title compound (1.1 g, 37% yield) (2:1 ratio of regioisomers) as a white solid. LCMS M/Z (M+H) 225.

Experimental manipulation of intermediates D and E

Step 1:

5-fluoro-2-methyl-2,3-dihydrophthalazine-1,4-dione, and

8-Fluoro-2-methyl-2,3-dihydrophthalazine-1,4-dione

A mixture of 4-fluoroisobenzofuran-1,3-dione (5.0 g, 30.10 mmol) and methylhydrazine (40% aqueous solution, 3.5 g, 30.10 mmol) in ethanol (100 mL) was heated at reflux for 15 h, at which point LCMS indicated the reaction was complete. The mixture was concentrated under reduced pressure to give a mixture of the title compounds (3.5 g, 60% yield) (2:1 ratio of regioisomers) as a yellow solid. The crude material was used in the next step without further purification. LCMS M/Z (M+H) 195.

Step 2:

4-chloro-5-fluoro-2-methylphthalazin-1(2H)-one, and

4-Chloro-8-fluoro-2-methylphthalazin-1(2H)-one

A mixture of 5- or 8-fluoro-2-methyl-2,3-dihydrophthalazine-1,4-dione (3.5 g, 18.03 mmol) in _POCl₃ (50 mL) was heated at reflux for 15 h, at which point LCMS indicated the reaction was complete. The mixture was concentrated under reduced pressure, and the residue was quenched by adding a saturated aqueous solution of _NaHCO₃ (100 mL), followed by extraction with ethyl acetate ( ₃ x 100 mL). The combined organic extracts were dried over _Na₂SO₄ and concentrated under reduced pressure. The residue was purified by silica gel chromatography (petroleum ether:ethyl acetate = 5:1) to give the title compound (1.2 g, 31% yield) (2:1 ratio of regioisomers) as a white solid. LCMS M/Z (M+H) 213.

Experimental operation of intermediate F

Step 1:

2-Methyl-2,3-dihydrophthalazine-1,4-dione

A mixture of isobenzofuran-1,3-dione (100 g, 675.15 mmol) and methylhydrazine (40% aqueous solution, 233.29 g, 2.03 mol) in methanol (1000 mL) was heated at reflux for 15 h, at which point LCMS indicated the reaction was complete. The mixture was concentrated under reduced pressure to give the crude title compound (103.1 g, 87% yield) as a yellow solid. The crude material was used in the next step without further purification. LCMS M/Z (M+H) 177.

Step 2:

4-Chloro-2-methylphthalazin-1(2H)-one

A mixture of 2-methyl-2,3-dihydrophthalazine-1,4-dione (100.0 g, 567.63 mmol) in _POCl₃ (1000 mL) was heated to reflux for 15 h, at which point LCMS indicated the reaction was complete. The mixture was concentrated under reduced pressure. The residue was quenched by adding a saturated aqueous solution of _NaHCO₃ (300 mL), followed by extraction with ethyl acetate ( ₃ x 300 mL). The combined organic extracts were dried over _Na₂SO₄ and concentrated under reduced pressure. The resulting residue was purified by silica gel chromatography (petroleum ether:ethyl acetate = 5:1) to give the title compound (15.6 g, 14% yield) as a white solid. LCMS M/Z (M+H) 195.

Experimental operation of intermediate G

Step 1:

4,6-Dichloropyridazin-3(2H)-one and 5,6-Dichloropyridazin-3(2H)-one

A solution of 3,4,6-trichloropyridazine (20.0 g, 109.04 mmol) in HOAc (100 mL) was heated at 100°C for 12 h, at which point TLC indicated the reaction was complete. The reaction mixture was concentrated under reduced pressure, and the resulting residue was purified by silica gel chromatography (petroleum ether:ethyl acetate = 1:1) to give the title compound (11.2 g, 63% yield) (1:1 ratio of regioisomers) as a white solid. LCMS M/Z (M+H) 165.

Step 2:

4,6-Dichloro-2-methylpyridazin-3(2H)-one and 5,6-Dichloro-2-methylpyridazin-3(2H)-one

To a suspension of 4,6-dichloropyridazin-3(2H)-one and 5,6-dichloropyridazin-3(2H)-one (11.2 g, 67.89 mmol) and Cs ₂ CO ₃ (33.2 g, 101.83 mmol) in DMF (100 mL) was added CH ₃ I (10.6 g, 74.68 mmol). The resulting mixture was stirred at 25 ° C for 15 h, at which time TLC indicated that the reaction was complete. The reaction mixture was quenched by adding a saturated aqueous solution of ammonium chloride (100 mL) and then extracted with ethyl acetate (3 x 100 mL). The combined organic extracts were dried over Na ₂ SO ₄ and concentrated under reduced pressure. The residue was purified by silica gel chromatography (petroleum ether:ethyl acetate = 1:1) to give the title compound (7.5 g, 62% yield) (1:1 ratio of regioisomers) as a white solid. LCMS M/Z (M+H) 179.

Step 3:

4-Amino-6-chloro-2-methylpyridazin-3(2H)-one

A mixture of 4,6-dichloro-2-methylpyridazin-3(2H)-one (7.5 g, 41.90 mmol, containing a mixture of 5,6-dichloro-2-methylpyridazin-3(2H)-one) and ammonium hydroxide (48%, 50 mL) was heated at 120°C in a sealed tube for 15 h, at which point LCMS indicated the reaction was complete. After cooling, the product was collected by filtration. The crude solid was washed with water and dried under reduced pressure to give a mixture of two regioisomers (3.8 g, 57%, 1:1 ratio) as a white solid. LCMS M/Z (M+H) 160.

Step 4:

4-Amino-6-chloro-5-iodo-2-methylpyridazin-3(2H)-one

1-iodopyrrolidine-2,5-dione (5.4 g, 23.81 mmol) was added to a solution of 4-amino-6-chloro-2-methylpyridazine-3 (2H) -one (3.8 g, 23.81 mmol, containing a mixture of regioisomers) in acetonitrile (60 mL). The reaction mixture was heated to reflux for 3 h, at which point LCMS indicated that the reaction was complete. After cooling, the reaction mixture was quenched by adding a saturated aqueous solution of ammonium chloride (20 mL) and then extracted with ethyl acetate (3 x 50 mL). The combined organic extracts were dried over Na ₂ SO ₄ and concentrated under reduced pressure. The residue was purified by silica gel chromatography (petroleum ether: ethyl acetate = 1: 3) to obtain a mixture of regioisomers in a 1: 1 ratio (2.5 g, 37% yield) as a yellow solid. LCMS M/Z (M+H) 286.

Step 5:

(E)-4-Amino-6-chloro-5-(2-ethoxyvinyl)-2-methylpyridazin-3(2H)-one

A mixture of 4-amino-6-chloro-5-iodo-2-methylpyridazin-3(2H)-one (2.50 g, 8.76 mmol, mixture of regioisomers), (E)-(2-ethoxyvinyl)boronic acid (1.22 g, 10.51 mmol), Cs ₂ CO ₃ (5.71 g, 17.51 mmol) and Pd(dppf)Cl ₂ (641 mg, 0.88 mmol) in dioxane/H ₂ O (50 mL) was heated at 100 ° C under an N ₂ atmosphere for 15 h, at which point LCMS indicated that the reaction was complete. The mixture was concentrated under reduced pressure and the residue was diluted with brine (60 mL). The solution was then extracted with ethyl acetate (3 x 50 mL). The combined organic extracts were dried over Na ₂ SO ₄ and concentrated under reduced pressure. The residue was purified by silica gel chromatography (petroleum ether:ethyl acetate=1:1) to give the title compound (800 mg, 40% yield) as a light yellow solid. LCMS M/Z (M+H) 230. Only the desired regioisomer was isolated at this step.

Step 6:

4-Chloro-6-methyl-1H-pyrazolo[2,3-d]pyridazin-7(6H)-one

A solution of (E)-4-amino-6-chloro-5-(2-ethoxyvinyl)-2-methylpyridazine-3(2H)-one (800 mg, 3.48 mmol) in HOAc (30 mL) was heated at reflux for 15 h, at which point LCMS indicated the reaction was complete. The mixture was concentrated under reduced pressure and the residue was purified by silica gel chromatography (petroleum ether:ethyl acetate=3:1) to give the title compound (500 mg, 78% yield) as a white solid. LCMS M/Z(M+H)184.

Experimental manipulation of intermediates H and I

Step 1:

2-methyl-2,3-dihydropyrido[3,4-d]pyridazine-1,4-dione, and

3-Methyl-2,3-dihydropyrido[3,4-d]pyridazine-1,4-dione

A mixture of dimethyl pyridine-3,4-dicarboxylate (5.0 g, 25.62 mmol) and methylhydrazine (40% aqueous solution, 8.85 g, 76.86 mmol) in ethanol (100 mL) was heated under reflux for 15 h, at which point LCMS indicated the reaction was complete. The mixture was concentrated under reduced pressure to give the title compound (4.5 g, 99% yield) (the ratio of the two isomers was 2:1) as a yellow solid. The crude material was used in the next step without further purification. LCMS M/Z (M+H) 178.

Step 2:

4-chloro-2-methylpyrido[3,4-d]pyridazin-1(2H)-one, and

1-Chloro-3-methylpyrido[3,4-d]pyridazin-4(3H)-one

A mixture of 2 or 3-methyl-2,3-dihydropyrido[3,4-d]pyridazine-1,4-dione (4.5 g, 25.4 mmol) in POCl ₃ (70 mL) was heated under reflux for 15 h, at which point LCMS indicated the reaction was complete. The mixture was concentrated under reduced pressure. The residue was quenched by adding a saturated aqueous solution of NaHCO ₃ (100 mL), followed by extraction with ethyl acetate (3 x 120 mL). The combined organic extracts were dried over Na ₂ SO ₄ and concentrated under reduced pressure. The resulting residue was purified by silica gel chromatography (petroleum ether: ethyl acetate = 2: 1) to give the title compound (1.5 g, 30% yield) (2: 1 ratio of regional isomers) as a white solid. LCMS M/Z(M+H)196.

Experimental manipulation of intermediate J

Step 1:

4-Chloro-2-ethylphthalazin-1(2H)-one

To a suspension of 4-chlorophthalazin-1(2H)-one (1.00 g, 5.54 mmol) and potassium carbonate (1.53 g, 11.1 mmol) in N,N-dimethylformamide (20 mL) was added iodoethane (1.04 g, 6.64 mmol). After addition, the reaction mixture was stirred at 80 ° C for 15 h and then poured into water (20 mL). The resulting precipitate was collected by filtration, washed with water, and dried under reduced pressure to give the title compound (0.8 g, 69% yield) as a brown solid. The crude material was used in the next step without further purification. LCMS M / Z (M + H) 209.

Experimental manipulation of intermediates K and L

Step 1:

2,5-dimethyl-2,3-dihydrophthalazine-1,4-dione, and

2,8-Dimethyl-2,3-dihydrophthalazine-1,4-dione

A mixture of 4-methylisobenzofuran-1,3-dione (5.0 g, 30.1 mmol) and 40% aqueous methylhydrazine solution (10.7 g, 92.5 mmol) in ethanol (100 mL) was heated at reflux for 15 h, at which point LCMS indicated the reaction was complete. The mixture was concentrated under reduced pressure to give the title compound (4.3 g, 73% yield) (2:1 ratio of regioisomers) as a yellow solid. The crude mixture was used in the next step without further purification. LCMS M/Z (M+H) 191.

Step 2:

4-Chloro-2,5-dimethylphthalazin-1(2H)-one and 4-chloro-2,8-dimethylphthalazin-1(2H)-one

A mixture of 2,5- or 8-dimethyl-2,3-dihydrophthalazine-1,4-dione (4.3 g, 22.6 mmol) in _POCl₃ (70 mL) was heated at reflux for 15 h, at which point LCMS indicated the reaction was complete. The mixture was concentrated under reduced pressure, and the residue was quenched by adding a saturated aqueous solution of _NaHCO₃ (100 mL), followed by extraction with ethyl acetate ( ₃ x 100 mL). The combined organic extracts were dried over _Na₂SO₄ and concentrated under reduced pressure. The residue was purified by silica gel chromatography (petroleum ether:ethyl acetate = 5:1) to give the title compound (1.5 g, 32% yield) (2:1 ratio of regioisomers) as a white solid. LCMS M/Z (M+H) 209.

Experimental operation of intermediate M

2-(But-2-en-1-yl)-4-chlorophthalazin-1(2H)-one

A round-bottom flask was charged with 4-chlorophthalazin-1-ol (0.75 g, 4.2 mmol), cesium carbonate (2.7 g, 8.3 mmol), and a stir bar. MeCN (20 mL) was added, followed by 1-chlorobut-2-ene (0.81 mL, 8.3 mmol), and the mixture was stirred at 50°C for 18 h. The reaction mixture was diluted with ethyl acetate, filtered, and celite added. The filtrate was evaporated in vacuo and purified by flash chromatography on silica gel (eluting with hexane and ethyl acetate). Concentration in vacuo afforded the title compound as a white crystalline solid. LCMS M/Z (M+H) 235.

Experimental operation of intermediate N

Step 1:

2-Methyl-2,3,5,6,7,8-hexahydrophthalazine-1,4-dione

A pyrex vial was charged with 4,5,6,7-tetrahydroisobenzofuran-1,3-dione (1.0 g, 6.6 mmol) and potassium acetate (0.77 g, 7.9 mmol). To this solid was added a 1:1 mixture of _H₂O :AcOH (10 mL) and methylhydrazine (40% in water, 0.91 g, 7.9 mmol). The vial was sealed and the reaction mixture was heated at 115°C overnight. After cooling to ambient temperature, the reaction mixture was concentrated to dryness with toluene (3X). LCMS M/Z (M+H) 181.

Step 2:

4-Chloro-2-methyl-5,6,7,8-tetrahydrophthalazin-1(2H)-one

In a 100 mL round-bottom flask, crude 2-methyl-2,3,5,6,7,8-hexahydrophthalazine-1,4-dione (1.18 g, 6.55 mmol) was dissolved in _POCl₃ (30 mL) and heated to 110°C overnight. The reaction mixture was cooled to ambient temperature, and then excess _POCl₃ was removed by rotary evaporation. The residue was slowly poured into a saturated aqueous solution of _NaHCO₃ and the pH was adjusted to approximately 8. The aqueous phase was extracted with dichloromethane (3X). The combined organic phases were dried _{over Na₂SO₄} , filtered, concentrated, and purified by silica gel chromatography using a 10-30% gradient of ethyl acetate/hexane as the eluent to afford the title compound (0.54 g, 41% yield) as a white solid. LCMS M/Z (M+H) 199.

Experimental manipulation of intermediate O

Step 1:

2-Methyl-2,3,6,7-tetrahydro-1H-cyclopenta[d]pyridazine-1,4(5H)-dione

A pyrex vial was charged with 5,6-dihydro-1H-cyclopenta[c]furan-1,3(4H)-dione (1.0 g, 7.2 mmol) and potassium acetate (0.85 g, 8.7 mmol). To this solid was added a 1:1 mixture of _H₂O :AcOH (10 mL) and methylhydrazine (0.46 mL, 8.7 mmol). The vial was sealed and the reaction mixture was heated at 115°C overnight. The reaction mixture was cooled to ambient temperature and concentrated to dryness with toluene (3X). LCMS M/Z (M+H) 167.

Step 2:

4-Chloro-2-methyl-2,5,6,7-tetrahydro-1H-cyclopenta[d]pyridazin-1-one

In a 100 mL round-bottom flask, crude 2-methyl-2,3,6,7-tetrahydro-1H-cyclopenta[d]pyridazine-1,4(5H)-dione (1.2 g, 7.1 mmol) was dissolved in _POCl₃ (30 mL) and heated to 110°C overnight. The reaction mixture was cooled to ambient temperature, and then excess _POCl₃ was removed by rotary evaporation. The residue was slowly poured into a saturated aqueous solution of _NaHCO₃ and the pH was adjusted to approximately 8. The aqueous phase was extracted with dichloromethane ( _3X ), and the combined organic phases were dried over _Na₂SO₄ , filtered, concentrated, and purified by silica gel chromatography using a 10-30% gradient of ethyl acetate/hexane as the eluent to give the title compound (0.86 g, 66%) as a brown solid. LCMS M/Z (M+H) 199.

Experimental operation of intermediate P

Step 1:

1,2,3,4-Tetrahydroquinoline-2-carboxylic acid

A mixture of quinoline-2-carboxylic acid (2.5 g, 14 mmol) and PtO ₂ (0.2 g) in methanol (50 mL) was stirred under hydrogen (15 psi) at 25° C. for 5 h, at which point TLC indicated the reaction was complete. The solid was removed by filtration and the filtrate was concentrated under reduced pressure to afford the crude title compound (1.5 g, 59% yield) as a yellow solid. LCMS M/Z (M+H) 178.

Step 2:

1-Methyl-1,2,3,4-tetrahydroquinoline-2-carboxylic acid methyl ester

To a stirred and cooled suspension of 1,2,3,4-tetrahydroquinoline-2-carboxylic acid (1.5 g, 8.5 mmol) and potassium carbonate (2.92 g, 21.2 mmol) in DMF (30 mL) was added iodomethane (3.0 g, 21 mmol). After addition, the resulting mixture was stirred at ambient temperature for 2 h, at which point LCMS indicated that the reaction was complete. The reaction mixture was poured into water (50 mL) and then extracted with ethyl acetate (3 x 30 mL). The combined organic extracts were dried over Na ₂ SO ₄ and concentrated under reduced pressure to give the title compound (0.6 g, 35% yield) as a red solid. The crude material was used in the next step without further purification. LCMS M/Z(M+H)206.

Step 3:

1-Methyl-1,2,3,4-tetrahydroquinoline-2-carboxamide

A mixture of methyl 1-methyl-1,2,3,4-tetrahydroquinoline-2-carboxylate (600 mg, 2.92 mmol) in 4M ammonia/methanol solution (20 mL) was heated at 80°C for 15 h, at which point LCMS indicated the reaction was complete. The solvent was evaporated under reduced pressure to give the crude title compound (500 mg, 90% yield) as a yellow solid. LCMS M/Z (M+H) 191.

Step 4:

(1-Methyl-1,2,3,4-tetrahydroquinolin-2-yl)methanamine

To a cooled (0 ° C) solution of 1-methyl-1,2,3,4-tetrahydroquinoline-2-carboxamide (0.50 g, 2.63 mmol) in tetrahydrofuran (50 mL) was added LiAlH ₄ (0.30 g, 7.9 mmol). After addition, the reaction mixture was heated at 60 ° C for 15 h, at which time LCMS indicated that the reaction was complete. The mixture was quenched by adding aqueous NaOH solution (10%, 0.3 mL) and water (1 mL) and filtered. The filtrate was evaporated under reduced pressure to give the crude title compound (0.20 g, 43% yield) as a yellow oil. LCMS M/Z (M+H) 177. The compound was separated into a racemic mixture.

Experimental operation of intermediate Q

Step 1:

8-(4-Chlorophenyl)-1,4-dioxa-8-azaspiro[4.5]decane

To a pyrex vial was added sodium 2-methylpropan-2-ol (2.24 g, 23.4 mmol), RuPhos 3rd generation precatalyst (0.26 g, 0.31 mmol), and 1-chloro-4-iodobenzene (5.57 g, 23.4 mmol). The vial was evacuated and purged with _N₂ (3×), followed by addition of toluene (10 mL) and 1,4-dioxa-8-azaspiro[4.5]decane (2.0 mL, 166 mmol). The vial was evacuated and purged with _N₂ (3×), then heated at 100°C overnight. The reaction mixture was cooled to ambient temperature, concentrated to dryness, and purified by silica gel chromatography (10% ethyl acetate/hexane) to afford 8-(4-chlorophenyl)-1,4-dioxa-8-azaspiro[4.5]decane (3.89 g, 98% yield). LCMS M/Z (M+H) 254.

Step 2:

1-(4-chlorophenyl)piperidin-4-one

In a 250mL round-bottom flask, 8-(4-chlorophenyl)-1,4-dioxa-8-azaspiro[4.5]decane (3.89g, 5.34mmol) was diluted in acetone (100mL), and then 6M aqueous hydrochloric acid solution (50mL) was adjusted and the reaction mixture was heated at reflux overnight. The reaction mixture was cooled and then concentrated to remove acetone. The aqueous solution was neutralized with 6M NaOH (50mL) and then extracted with ethyl acetate (3X). The organic layers were combined, washed with brine, dried over Na ₂ SO ₄ , filtered and concentrated. The crude residue was purified by silica gel chromatography (10% ethyl acetate/hexane) to obtain 1-(4-chlorophenyl)piperidin-4-one (2.96g, 92% yield) as a yellow solid. LCMS M/Z(M+H)210.

Step 3:

3-Bromo-1-(4-chlorophenyl)piperidin-4-one

In a 50 mL round-bottom flask, under an _N atmosphere, 1-(4-chlorophenyl)piperidin-4-one (0.500 g, 2.39 mmol) was diluted in dichloromethane (12 mL) and cooled to -10 ° C (salt/ice bath). Bromine (0.12 mL, 2.4 mmol) was added dropwise over 3 minutes, maintaining the internal reaction temperature at -2 to -4 ° C. The reaction mixture was then warmed to ambient temperature and mixed until clear. A saturated aqueous solution of NaHCO ₃ was added and the reaction mixture was stirred for 0.5 h. The layers were separated and the aqueous layer was extracted with dichloromethane (1X). The combined organic layers were dried and filtered over Na ₂ SO ₄ , concentrated and used directly in subsequent reactions. LCMS M/Z (M+H) 288, 290.

Step 4:

1-(4-Chlorophenyl)-3-(dimethylamino)piperidin-4-one

In a 100 mL round-bottom flask, crude 3-bromo-1-(4-chlorophenyl)piperidin-4-one (688 mg, 2.39 mmol) was dissolved in tetrahydrofuran (20 mL) and cooled to 0°C. Dimethylamine (2 M in tetrahydrofuran, 4.78 mL, 9.56 mmol) was then added and the reaction mixture was allowed to slowly warm to ambient temperature overnight. The crude reaction mixture was concentrated and purified by silica gel chromatography to afford the title compound (212 mg, 35% yield) as an oil. LCMS M/Z (M+H) 253.

Step 5:

1-(4-Chlorophenyl)-3-(dimethylamino)piperidin-4-one oxime

In a 25 mL round-bottom flask, 1-(4-chlorophenyl)-3-(dimethylamino)piperidin-4-one (0.21 g, 0.84 mmol) and hydroxylamine hydrochloride (0.117 g, 1.68 mmol) were dissolved in ethanol (8 mL). Pyridine (0.22 mL, 2.72 mmol) was added and the reaction mixture was heated at 80°C overnight. The reaction mixture was concentrated to dryness and purified by silica gel chromatography (0-100% gradient of 90:10:1 dichloromethane:methanol: _NH₄OH mixed with dichloromethane) to give the title compound (0.18 g, 80% yield) as a yellow oil. LCMS M/Z (M+H) 268.

Step 6:

1-(4-Chlorophenyl)-N3,N3-dimethylpiperidine-3,4-diamine

In a 50 mL round-bottom flask, (E)-1-(4-chlorophenyl)-3-(dimethylamino)piperidin-4-one oxime (0.18 g, 0.68 mmol) was diluted with tetrahydrofuran (5 mL). The reaction flask was evacuated and purged with _N2 (3X). Borane (2M in tetrahydrofuran, 1.35 mL, 1.35 mmol) was then added and the reaction mixture was heated at 70 ° C overnight. The reaction mixture was cooled to ambient temperature, quenched by adding methanol (6 mL), and then heated at 70 ° C for 1 h. The reaction mixture was concentrated to dryness and purified by silica gel chromatography (50% dichloromethane and 50% 90:10:1 dichloromethane:methanol: _NH4OH ) to give the title compound (0.071 g, 42% yield) as a colorless oil. LCMS M/Z (M+H) 254. The compound was separated into a racemic mixture of diastereomers.

Experimental operation of intermediate R

1-(2-Chlorophenyl)-N1,N1-dimethylethylene-1,2-diamine

The title compound was prepared from 2-chlorophenylacetonitrile according to the procedure described in patent WO1984003278. LCMS M/Z (M+H) 199. The compound was isolated as a racemic mixture.

Experimental manipulation of intermediates S and T

The following intermediates were prepared in a similar manner to Intermediate R using the appropriate starting material in place of 2-chlorophenylacetonitrile.

Experimental manipulation of intermediate U

(1R,2R)-1-Dimethylamino-2-indanol

Formic acid (90% purity, 1.2 mL) and 37% aqueous formaldehyde (9 mL) were added to (1R,2R)-1-amino-2-indanol (1.0 g, 6.7 mmol). After heating to 80°C for 3 h, the solution was cooled to room temperature and treated with a saturated aqueous solution _{of Na₂CO₃} to make it alkaline. The mixture was extracted twice with ethyl acetate, and _the combined organic layers were washed with brine, dried over _Na₂SO₄ , filtered, and concentrated under reduced pressure. Purification: 40 g silica gel column using 1:10:90 _NH₄OH :methanol:dichloromethane/dichloromethane, 20% to 100% over 6 column volumes. Pure fractions were concentrated in vacuo to yield the title intermediate (0.96 g, 81% yield). LCMSM/Z (M+H) 178. The compound was isolated as a single enantiomer.

(1R,2S)-2-Azido-N,N-dimethyl-2,3-dihydro-1H-inden-1-amine

(1R,2R)-1-(Dimethylamino)-2,3-dihydro-1H-inden-2-ol (574 mg, 3.22 mmol) was dissolved in tetrahydrofuran (20 mL). Triphenylphosphine (1.1 g, 4.2 mmol) was added and dissolved, and the mixture was cooled to 0°C. DIAD (970 μl, 4.83 mmol) was added, and the mixture was stirred for 15 minutes, followed by DPPA (830 μl, 3.86 mmol). The reaction was stirred for 3 hours, and the reaction was complete. The crude reaction mixture was applied to silica gel and purified by column chromatography (1:10:90 _NH₄OH :methanol:dichloromethane, 0% to 100% in dichloromethane). Triphenylphosphine oxide co-eluted with the product. The title intermediate was obtained after vacuum concentration. LCMS M/Z (M+H) 203. The compound was isolated as a single enantiomer.

(1R,2S)-N1,N1-Dimethyl-2,3-dihydro-1H-indene-1,2-diamine

(1R,2S)-2-Azido-N,N-dimethyl-2,3-dihydro-1H-indene-1-amine (645 mg, 3.21 mmol, estimated) was dissolved in ethyl acetate (5 mL), the reaction flask was degassed using nitrogen/vacuum cycles, and 5% wet Pd/charcoal (360 mg) was added. The flask was stoppered with a rubber stopper and placed under a hydrogen atmosphere using vacuum/ _H2 cycles. Stirred for 60 hours. The reaction was complete. Filtered on Celite, the palladium was washed with ethyl acetate. Evaporated under high vacuum. The product was used immediately in the next step. (1R,2S)-N1,N1-dimethyl-2,3-dihydro-1H-indene-1,2-diamine (457 mg, 75% yield over 2 steps). LCMS M/Z (M+H) 177. The compound was isolated as a single enantiomer.

Experimental manipulation of intermediate V

The title compound was prepared in an analogous manner to Intermediate U using (1S,2S)-1-dimethylamino-2-indanol as the starting material instead of (1R,2R)-1-dimethylamino-2-indanol. The compound was isolated as a single enantiomer.

Synthesis of Example Compounds

Example 1A/B

8-((2-(Dimethylamino)-2-phenylethyl)amino)-6-methylpyrido[2,3-d]pyridazin-5(6H)-one

Under microwave conditions, a mixture of 8-chloro-6-methylpyrido[2,3-d]pyridazin-5(6H)-one (Intermediate A, 100 mg, 0.510 mmol), N ¹ ,N ¹ -dimethyl-1-phenylethane-1,2-diamine (100 mg, 0.610 mmol), tert-butyl-XPhos (44 mg, 0.10 mmol), t-BuOK (115 mg, 1.02 mmol) and Pd ₂ dba ₃ (47 mg, 0.050 mmol) in DMF (5 mL) was heated at 100 ° C. under N ₂ for 2 h, at which time LCMS indicated that the reaction was complete. The reaction mixture was poured into water (5 mL) and then extracted with ethyl acetate (3 x 10 mL). The combined organic extracts were dried over Na ₂ SO ₄ and concentrated under reduced pressure. The residue was purified by reverse phase chromatography (30-60% acetonitrile/0.1% NH ₄ OH/water) to provide the title compound (24 mg, 14% yield) as a white solid. ¹ H NMR (400 MHz, Methanol-d 4 ) δ 8.99-8.97 (m, 1H), 8.63-8.61 (m, 1H), 7.72-7.79 (m, 1H), 7.37-7.32 (m, 5H), 4.01-3.97 (m, 1H), 3.88-3.71 (m, 5H), 2.29 (s, 6H). LCMS M/Z (M+H) 324.

(S)-8-((2-(dimethylamino)-2-phenylethyl)amino)-6-methylpyrido[2,3-d]pyridazin-5(6H)-one and (R)-8-((2-(dimethylamino)-2-phenylethyl)amino)-6-methylpyrido[2,3-d]pyridazin-5(6H)-one

The enantiomers were separated by SFC using the conditions described in the next section. (Method C: Chiralpak ID column, 5 μM particle size, 2 x 10 cm dimensions, 75% CO ₂ /25% iPrOH containing 0.1% NH ₄ OH eluent, 70 mL/min flow rate, 100 bar pressure, 40° C.).

Example 2-18

In a similar manner to Example 1, the following compounds were prepared using the corresponding intermediates: Intermediate A (Examples 2, 7, 8, 11, 14-16 and 18), Intermediate B (Example 3), Intermediate C (Example 4), Intermediate D (Example 5), Intermediate E (Example 6), Intermediate F (Example 9), Intermediate G (Example 10), Intermediate H (Example 12), Intermediate I (Example 13) or Intermediate J (Example 17A/B). All examples in the table below were prepared using commercially available amines. Specific conditions for separating enantiomers are reported in the following section.

Example 19

Step 1

tert-Butyl 3-((6-methyl-5-oxo-5,6-dihydropyrido[2,3-d]pyridazin-8-yl)amino)azetidine-1-carboxylate

Under microwave conditions, a mixture of 8-chloro-6-methylpyrido [2,3-d] pyridazine-5 (6H) -one (intermediate C, 300 mg, 1.53 mmol), 3-aminoazetidine-1-carboxylic acid tert-butyl ester (318 mg, 1.83 mmol), tert-butyl-XPhos (132 mg, 0.300 mmol), t-BuOK (345 mg, 3.06 mmol) and Pd ₂ dba ₃ (141 mg, 0.150 mmol) in DMF (8 mL) was heated at 100 ° C under N ₂ for 2 h, when LCMS indicated that the reaction had completed. The reaction mixture was poured into water (10 mL) and then extracted with ethyl acetate (3 x 10 mL). The combined organic extracts were dried over Na ₂ SO ₄ and concentrated under reduced pressure. The residue was purified by silica gel chromatography (petroleum ether/ethyl acetate=1: 1) to give the title compound (220 mg, 43% yield) as a brown solid. LCMS M/Z (M+H) 332.

Step 2

8-(Azetidin-3-ylamino)-6-methylpyrido[2,3-d]pyridazin-5(6H)-one

To a solution of tert-butyl 3-((6-methyl-5-oxo-5,6-dihydropyrido[2,3-d]pyridazin-8-yl)amino)azetidine-1-carboxylate (100 mg, 0.300 mmol) in methanol (3 mL) was added 2 M hydrogen chloride/methanol (3 mL). The resulting mixture was stirred at ambient temperature for 30 min and then concentrated in vacuo. The residue was purified by reverse phase chromatography (acetonitrile 28-58%/0.1% NH ₄ OH/water) to give the title compound (8.2 mg, 12% yield) as a white solid. 1H NMR (400 MHz, methanol-d4) δ 9.06-9.04 (m, 1H), 8.64-8.62 (m, 1H), 7.84-7.81 (m, 1H), 4.79-4.77 (m, 1H), 4.08-4.03 (m, 2H), 3.95-3.91 (m, 2H), 3.68 (s, 3H). LCMS M/Z (M+H) 232.

Example 20

4-((2-(dimethylamino)-2-(4-fluorophenyl)ethyl)amino)-2-methylphthalazin-1(2H)-one

A mixture of 4-chloro-2-methylphthalazin-1(2H)-one (Intermediate F, 100 mg, 0.515 mmol), 1-(4-fluorophenyl)-N1,N1-dimethylethane-1,2-diamine (180 mg, 1.04 mmol), sodium 2-methylpropan-2-ol (100 mg, 1.04 mmol), and RuPhos 3rd generation precatalyst (43 mg, 0.050 mmol) in toluene (5 mL) was degassed. The resulting mixture was heated at 110°C for 2 h. The mixture was cooled and extracted with ethyl acetate and water. The organic layer was separated, dried _{over Na2SO4} , filtered, concentrated in vacuo, and purified on a 10 g biotage silica gel column using ethyl acetate to 5% methanol/ethyl acetate to afford the title compound (70 mg, 40% yield). ^1H NMR (400 MHz, acetone-d6) δ 8.32-8.35 (m, 1H), 7.77-7.88 (m, 3H), 7.41-7.45 (m, 2H), 7.11 (t, J = 8.79 Hz, 2H), 5.80-5.85 (m, 1H), 3.81-3.88 (m, 2H), 3.61 (s, 3H), 3.52-3.58 (m, 1H), 2.23 (s, 6H). LCMS M/Z (M+H) 341. The compound was isolated as a racemic mixture.

Examples 21-47

The following compounds were prepared in a manner similar to Example 20 using the corresponding intermediates: Intermediate F (Examples 21-30, 33-35, 38-45, and 47), Intermediate M (Example 31), Intermediate N (Example 36), Intermediate O (Example 37), or Intermediate A (Examples 32 and 46). For Examples 25-27, 32-34, and 46-47, the diamine starting material was prepared according to the previously described procedures (Intermediates R, S, T, Q, U, and V). All other examples were prepared using commercially available diamines. Specific conditions for separation of enantiomers are reported in the following section.

Example 48

The following compounds were prepared in a similar manner to Example 20 using t-BuRockPhos 3rd generation precatalyst instead of RuPhos 3rd generation precatalyst.

Example 49

Step 1:

4-((2,2-diethoxyethyl)amino)-2-methylphthalazin-1(2H)-one

The title intermediate was prepared in a manner similar to Example 20, but heating at 130° C. instead of 110° C. LCMSM/Z (M+H) 292.

Step 2:

2-((3-methyl-4-oxo-3,4-dihydrophthalazin-1-yl)amino)acetaldehyde

To a suspension of 4-((2,2-diethoxyethyl)amino)-2-methylphthalazin-1(2H)-one (75 mg, 0.26 mmol) in water (1 mL) was added 1.25 M hydrogen chloride/ethanol (5.1 mL, 6.4 mmol) at room temperature. The reaction mixture was heated at 40°C for 45 min and then concentrated in vacuo. The crude product was used in the next step without purification. LCMS M/Z (M+H) 218.

Step 3:

4-((2-((2-methoxyethyl)(methyl)amino)ethyl)amino)-2-methylphthalazin-1(2H)-one

To a solution of crude 2-((3-methyl-4-oxo-3,4-dihydrophthalazin-1-yl)amino)acetaldehyde in methanol (3 mL) was added 2-methoxy-N-methylethylamine (0.0688 g, 0.7722 mmol) and sodium cyanoborohydride (0.024 g, 0.39 mmol). The pH was then adjusted to 7 with acetic acid, and the reaction mixture was stirred for 30 minutes before being quenched with aqueous sodium bicarbonate. The product was extracted with ethyl acetate (repeated 3 times), and _the combined organic layers were dried over _Na₂SO₄ , filtered, and concentrated to dryness under reduced pressure. The crude product was purified by flash chromatography (hexane/ethyl acetate 9:1 to 0:10) to give the title compound (22 mg, 29% yield). ¹ H NMR (400MHz, DMSO-d6) δ8.24(dd,J＝1.53,7.88Hz,1H),8.07(d,J＝7.88Hz,1H),7.88(dt,J＝1.42,7.61Hz,1H),7.81(dt,J＝1.00,7.60Hz,1H), 6.69(br.s,1H),3.54-3.59(m,3H),3.46(t,J＝5.80Hz,2H),3.35-3.41 (m,2H),3.22(s,3H),2.60-2.80(m,2H),2.35(br.s.,2H),1.90(s,3H). LCMS M/Z(M+H)291.

Examples 50-51

The following compounds were prepared in a manner similar to Example 49. All examples in the table below were prepared using commercially available amines.

Example 52

Step 1:

4-(((1R,2R)-2-aminocyclohexyl)amino)-2-methylphthalazin-1(2H)-one

In a pyrex vial, 4-chloro-2-methylphthalazin-1(2H)-one (300 mg, 1.54 mmol), RuPhos 3rd generation precatalyst (121 mg, 0.154 mmol) and sodium tert-butoxide (444 mg, 4.62 mmol) were added, the vial was sealed, the atmosphere was evacuated and purged with _N2 (3X). A solution of (+/-)-trans-cyclohexanediamine (370 μL, 3.08 mmol) in toluene (6 mL) was then added, and the reaction mixture was heated at 70°C overnight. The crude reaction mixture was concentrated in vacuo, deposited onto silica gel with methanol, and purified by silica gel chromatography using 85% 90:10:1 dichloromethane:methanol: _NH4OH as eluent. The product fractions were concentrated and lyophilized to give the title compound (191 mg, 45% yield). LCMS M/Z (M+H) 273.

Step 2:

4-(((1R,2R)-2-(dimethylamino)cyclohexyl)amino)-2-methylphthalazin-1(2H)-one

In a 25 mL round-bottom flask, 4-(((1R,2R)-2-aminocyclohexyl)amino)-2-methylphthalazin-1(2H)-one (129 mg, 0.48 mmol) was dissolved in methanol (5 mL). Formaldehyde (37% in water, 2 mL) and formic acid (88%, 2 mL) were added, and the reaction mixture was heated at 90°C overnight. The reaction mixture was cooled to ambient temperature, poured into a saturated aqueous solution _{of NaHCO₃, and extracted with dichloromethane. The extract was washed with brine, dried over Na₂SO₄ , filtered} , concentrated in vacuo, and purified by silica gel chromatography (eluting with 75% dichloromethane in a 90:10:1 mixture of dichloromethane:methanol: _NH₄OH ) to provide 4-(((1R,2R)-2-(dimethylamino)cyclohexyl)amino)-2-methylphthalazin-1(2H)-one (96 mg, 69% yield) as a white solid. ¹ H NMR (400MHz, DMSO-d6) δ8.24(dd,J＝1.4,7.8Hz,1H),8.01(d,J＝7.9Hz,1H),7.88(dt,J＝1.4,7.6Hz,1H),7.84-7.78(m,1H),6.18(d,J＝4.8Hz,1H),3. 60(t,J＝4.4Hz,1H),3.56(s,3H),2.54(d,J＝3.3Hz,1H),2.39(d,J＝12.5Hz ,1H),2.19(s,6H),1.87-1.75(m,2H),1.65(br.S.,1H),1.32-1.09(m,4H). LCMS M/Z(M+H)301. (racemic mixture; single diastereomer; and unknown relative stereochemistry)

Examples 53-54

The following compounds were prepared in a manner similar to Example 52. All examples in the table below were prepared using commercially available amines. Specific conditions for separation of enantiomers are reported in the following section.

Example 55

Step 1:

1-(2-Nitroprop-1-en-1-yl)-4-(phenylsulfonyl)benzene

A mixture of 4-(phenylsulfonyl)benzaldehyde (1.00 g, 4.06 mmol), nitroethane (3.05 g, 40.6 mmol), and ammonium acetate (0.08 g, 1 mmol) was heated at 120°C for 2 h. The resulting mixture was cooled, concentrated in vacuo, and purified on a 40 g biotage silica gel column using hexanes to 30% ethyl acetate/hexanes to afford the title compound (360 mg, 30% yield). LCMS M/Z (M+Na) 326. E/Z stereochemistry not determined.

Step 2:

(1S,2S)-N,N-Dimethyl-2-nitro-1-(4-(phenylsulfonyl)phenyl)propan-1-amine

To 1-(2-nitroprop-1-en-1-yl)-4-(phenylsulfonyl)benzene (0.360 g, 1.19 mmol) was added a 2M solution of dimethylamine in tetrahydrofuran (0.72 mL, 1.4 mmol). After completion of the reaction, the reaction mixture was concentrated in vacuo and co-evaporated with methanol to afford the title compound (360 mg, 87% yield) as a white solid. Since the compound was unstable, it was used immediately in the next step. LCMS M/Z (M+H) 349. The compound was isolated as a racemic mixture of unknown relative stereochemistry: assignments were made based on the most likely scenario, as described in the literature.

Step 3:

(1S,2S)-N1,N1-Dimethyl-1-(4-(phenylsulfonyl)phenyl)propane-1,2-diamine

To a solution of (1S,2S)-N,N-dimethyl-2-nitro-1-(4-(phenylsulfonyl)phenyl)propan-1-amine (0.360 g, 1.03 mmol) in methanol (10 mL) was added palladium hydroxide (0.140 g, 1.03 mmol). The resulting mixture was stirred under 1 atmosphere of hydrogen for 18 h. The mixture was filtered through celite and the filtrate was concentrated to dryness in vacuo. The residue was used as is in the next step. LCMS M/Z (M+H) 319. The compound was isolated as a racemic mixture with unknown relative stereochemistry: assignment was made based on the forward step.

Step 4:

4-(((1S,2S)-1-(dimethylamino)-1-(4-(phenylsulfonyl)phenyl)propan-2-yl)amino)-2-methylphthalazin-1(2H)-one

The title compound was prepared in a manner similar to Example 20. ¹ H NMR (400 MHz, acetone-d6) δ 8.35 (dd, J = 2.47, 6.68 Hz, 1H), 8.01 (dd, J = 7.78, 10.71 Hz, 3H), 7.86-7.93 (m, 2H), 7.81-7.85 (m, 2H), 7.61-7.73 (m, 3H), 7.56 (d, J = 8.24 Hz, 2H), 5.97 (d, J = 2.93 Hz, 1H), 4.28-4.58 (m, 1H), 3.72-3.82 (m, 1H), 3.63 (s, 3H), 2.17 (s, 6H), 1.08 (d, J = 6.23 Hz, 3H). LCMS M/Z (M+H) 477. The compound was isolated as a racemic mixture of unknown relative stereochemistry: assignments were made based on the most likely scenario according to the literature.

Examples 56-67

The following compounds were prepared using the corresponding intermediates in a similar manner to Example 55: Intermediate F (Examples 56-57 and 59-63) or Intermediate A (Examples 58 and 64-67). Specific conditions for separation of enantiomers are reported in the following section.

Example 68

Step 1:

Methyl 4-(2-nitroprop-1-en-1-yl)benzoate

A mixture of methyl 4-formylbenzoate (10.0 g, 60.92 mmol) and ammonium acetate (0.90 g, 12 mmol) in nitroethane (100 mL) was heated at 100 ° C for 15 h, at which point LCMS indicated that the reaction was complete. The solvent was concentrated in vacuo. The residue was dissolved in ethyl acetate (150 mL) and washed with water (2 x 100 mL). The separated organic layer was dried over Na ₂ SO ₄ and concentrated under reduced pressure to give the title compound (10.5 g, 78% yield) as a yellow solid. The crude material was used in the next step without further purification. The E/Z stereochemistry was not determined.

Step 2:

Methyl 4-((1S,2S)-1-(dimethylamino)-2-nitropropyl)benzoate

To a mixture of (Z)-methyl 4-(2-nitroprop-1-en-1-yl)benzoate (10.5 g, 47.5 mmol) in tetrahydrofuran (15 mL) was added a 2M solution of dimethylamine in tetrahydrofuran (118.7 mL, 237.3 mmol). After addition, the mixture was stirred at 25 ° C for 15 h, at which time LCMS indicated that the reaction was complete. The solvent was evaporated under reduced pressure to give the crude title compound (10.6 g, 84% yield) as a yellow solid. LCMS M / Z (M + H) 267. The compound was isolated as a racemic mixture with unknown relative stereochemistry: according to the literature, the assignment was based on the most likely scenario.

Step 3:

Methyl 4-((1S,2S)-2-amino-1-(dimethylamino)propyl)benzoate

A mixture of methyl 4-((1S,2S)-1-(dimethylamino)-2-nitropropyl)benzoate (10.0 g, 37.5 mmol) and Raney nickel (5.0 g) in methanol (50 mL) was stirred at 25 ° C. under hydrogen (15 psi) for 5 h, at which point LCMS indicated that the reaction was complete. The solid was removed by filtration, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel chromatography (petroleum ether: ethyl acetate = 1: 1) to give the title compound (8.6 g, 97% yield) as a colorless oil. LCMS M/Z (M+H) 237. The compound was separated into a racemic mixture with unknown relative stereochemistry: attributed based on the forward step.

Step 4:

Methyl 4-((1S,2S)-1-(dimethylamino)-2-((6-methyl-5-oxo-5,6-dihydropyrido[2,3-d]pyridazin-8-yl)amino)propyl)benzoate

In 20 parallel batches, a mixture of 8-chloro-6-methylpyrido[2,3-d]pyridazin-5(6H)-one (Intermediate A, 300 mg, 1.53 mmol), methyl 4-((1S,2S)-2-amino-1-(dimethylamino)propyl)benzoate (543 mg, 2.30 mmol), t-BuOK (344 mg, 3.07 mmol) and RuPhos 3rd generation precatalyst (30 mg, 0.04 mmol) in DMF (8 mL) was heated at 100 ° C. under N ₂ for 2 h, at which time LCMS indicated that the reaction was complete. The reaction mixture was poured into water (5 mL) and then extracted with ethyl acetate (3 x 10 mL). The combined organic extracts were dried over Na ₂ SO ₄ and concentrated in vacuo. The combined residues were purified by reverse phase chromatography (acetonitrile 50-80%/0.1% _NH4OH /water) to give the title compound (750 mg, 6% yield after combining 20 batches) as a white solid.1H NMR ( ⁴⁰⁰ MHz, Methanol-d4) δ 9.02-9.00 (m, 1H), 8.61-8.59 (m, 1H), 8.02 (d, J = 8.4 Hz, 2H), 7.86-7.89 (m, 1H), 7.39 (d, J = 8.0 Hz, 2H), 4.51-4.48 (m, 1H), 3.91 (s, 3H), 3.73 (s, 3H), 3.72-3.70 (m, 1H), 2.18 (s, 6H), 1.13 (d, J = 6.0 Hz, 3H). LCMS M/Z (M+H) 396. The compound was isolated as a racemic mixture of unknown relative stereochemistry: assignments were made based on the most likely scenario according to literature.

Examples 69-75

The following compounds were prepared using the corresponding intermediates in a manner similar to Example 68: Intermediate L (Example 69), Intermediate K (Example 70), Intermediate G (Example 71), Intermediate F (Example 73), or Intermediate A (Examples 72 and 74-75). For Examples 69-71 and 73-74, the diamine starting material was also prepared according to the procedures described above in Steps 1-3 of Example 55. Specific conditions for separating enantiomers are reported in the following section.

Example 76

Step 1

4-(2-Nitroprop-1-en-1-yl)phenol

The title intermediate was prepared in an analogous manner to Example 55, Step 1, using 4-hydroxybenzaldehyde instead of 4-(phenylsulfonyl)benzaldehyde. LCMS M/Z (M+H) 180. E/Z stereochemistry not determined.

Step 2

1-(Methoxymethoxy)-4-(2-nitroprop-1-en-1-yl)benzene

To a solution of 4-(2-nitrovinyl)phenol (13.5 g, 81.8 mmol) in acetonitrile (160 mL) maintained at room temperature using a water bath was added chloro(methoxy)methane (6.83 mL, 89.9 mmol), followed by portionwise addition of cesium carbonate (29.3 g, 89.9 mmol) (Caution: Exothermic reaction). After 1 h at room temperature, additional chloro(methoxy)methane (0.62 mL, 8.2 mmol) was added and the reaction mixture was stirred for an additional 2 h. When TLC showed complete consumption of the starting material, the reaction mixture was diluted with tert-butyl methyl ether (200 mL) and filtered through celite. The filtrate was concentrated under reduced pressure to dryness to give an orange-brown oil, which was used in the next step without purification. LCMS M/Z (M+H) 223.

Step 3

(1S,2S)-1-(4-(Methoxymethoxy)phenyl)-N,N-dimethyl-2-nitropropan-1-amine

To crude 1-(methoxymethoxy)-4-(2-nitroprop-1-en-1-yl)benzene (8.0 g, 36 mmol) was added 2M dimethylamine in tetrahydrofuran (89.6 mL, 179.2 mmol). After addition, the mixture was stirred at room temperature overnight, and the solvent was then evaporated in vacuo to yield the crude title compound. LCMS M/Z (M+H) 269.

Step 4

(1S,2S)-1-(4-(methoxymethoxy)phenyl)-N1,N1-dimethylpropane-1,2-diamine

The title intermediate was prepared in a similar manner to Example 55, Step 3. LCMS M/Z (M+H) 239.

Step 5

4-(((1S,2S)-1-(dimethylamino)-1-(4-(methoxymethoxy)phenyl)propan-2-yl)amino)-2-methylphthalazin-1(2H)-one

The title intermediate was prepared in a similar manner to Example 20. LCMS M/Z (M+H) 397.

Step 6

4-(((1S,2S)-1-(dimethylamino)-1-(4-hydroxyphenyl)propan-2-yl)amino)-2-methylphthalazin-1(2H)-one

To a solution of 4-(((1S,2S)-1-(dimethylamino)-1-(4-(methoxymethoxy)phenyl)propan-2-yl)amino)-2-methylphthalazin-1(2H)-one (1.5 g, 3.8 mmol) (low purity) in methanol (100 mL) was added 2M aqueous hydrochloric acid solution (50 mL) at room temperature. The mixture was heated to 45°C for 5 hours and then concentrated under reduced pressure. The pH of the aqueous solution was adjusted to 8, and the product was extracted with ethyl acetate (repeated twice). The combined organic layers were dried over _Na2SO4 , filtered, and concentrated to dryness under reduced pressure. The crude product was purified by flash chromatography (dichloromethane/methanol 97:3 to 60:40) _to give the title compound (650 mg, 48% yield). ¹ H NMR (400MHz, DMSO-d6) δ9.35 (s, 1H), 8.27 (dd, J = 1.31, 7.88Hz, 1H), 7.99 (d, J＝8.00Hz,1H),7.90(dt,J＝1.42,7.61Hz,1H),7.81-7.86(m,1H),7.03(d,J＝ 8.54Hz,2H),6.76(d,J＝8.54Hz,2H),6.31(d,J＝4.16Hz,1H),4.19-4.31(m,1 H), 3.61 (s, 3H), 3.51 (d, J = 9.63Hz, 1H), 2.05 (s, 6H), 1.02 (d, J = 6.13Hz, 3H). LCMS M/Z (M+H) 353. The compound was isolated as a racemic mixture of unknown relative stereochemistry: assignments were made based on the most likely scenario according to the literature.

Example 77

4-((1-(4-(Benzyloxy)phenyl)-1-(dimethylamino)propan-2-yl)amino)-2-methylphthalazin-1(2H)-one

To a solution of 1-(dimethylamino)-1-(4-hydroxyphenyl)propan-2-yl)amino)-2-methylphthalazin-1(2h)-one (20 mg, 0.06 mmol) in tetrahydrofuran (1 mL) at room temperature were added sodium 2-methylpropan-2-ol (6.5 mg, 0.07 mmol) and benzyl bromide (8 μL, 0.06 mmol). Additional sodium 2-methylpropan-2-ol (26 mg, 0.28 mmol) and benzyl bromide (32 μL, 0.24 mmol) were added over a period of several hours. After an 18-hour period, the resulting mixture was extracted with ethyl acetate and a saturated solution of ammonium chloride. The organic phase was separated and concentrated in vacuo, and the residue was purified by flash chromatography from ethyl acetate to 20% methanol/ethyl acetate to give the title compound (10 mg, 40% yield). ¹ H NMR (400MHz, acetone-d6) δ8.36(d,J＝7.32Hz,1H),7.77-7.98(m,3H),7.52(d,J＝7.32Hz,2H),7.42(t,J＝7.32Hz,2H),7.36(d,J＝7.32Hz,1H),7.28(d, J＝8.42Hz,2H),7.08(d,J＝8.61Hz,2H),6.25-6.35(m,1H),5.14(s,2H),4 .30(br.s.,1H),3.54-3.75(m,3H),2.17(s,6H),1.12(d,J＝6.04Hz,3H). LCMS M/Z(M+H)443. The compound was isolated as a racemic mixture of unknown relative stereochemistry: assignments were made based on the most likely scenario according to the literature.

Examples 78-80

The following compound was prepared in a similar manner to Example 77 using an alkyl halide instead of benzyl bromide.

Example 81

4-((1S,2S)-1-(Dimethylamino)-2-((6-methyl-5-oxo-5,6-dihydropyrido[2,3-d]pyridazin-8-yl)amino)propyl)benzoic acid

To a stirred solution of methyl 4-((1S,2S)-1-(dimethylamino)-2-((6-methyl-5-oxo-5,6-dihydropyrido[2,3-d]pyridazin-8-yl)amino)propyl)benzoate (700 mg, 0.77 mmol) in tetrahydrofuran (10 mL)/water (5 mL) was added lithium hydroxide (212 mg, 8.55 mmol) in portions. After addition, the mixture was stirred at 80 ° C for 2 h, at which time LCMS indicated that the reaction was complete. After cooling, the mixture was concentrated under reduced pressure. The residue was dissolved in water (5 mL) and acidified to pH 3-4 with 5 M aqueous hydrochloric acid solution. The resulting precipitate was collected by filtration, washed with water, and dried under reduced pressure to give the title compound (400 mg, 59% yield) as a white solid. 1H NMR (400 MHz, methanol-d4) δ 9.11-9.10 (m, 1H), 8.72-8.69 (m, 1H), 8.11 (d, J = 7.6 Hz, 2H), 7.91-7.89 (m, 1H), 7.58 (d, J = 8.0 Hz, 2H), 4.61-4.58 (m, 1H), 4.40-4.37 (m, 1H), 3.81 (s, 3H), 2.63 (s, 6H), 1.18 (d, J = 6.0 Hz, 3H). LCMS M/Z (M+H) 382. The compound was isolated as a racemic mixture of unknown relative stereochemistry: assignment based on Example 68.

Example 82

8-(((1S,2S)-1-(dimethylamino)-1-(4-(morpholine-4-carbonyl)phenyl)propan-2-yl)amino)-6-methylpyrido[2,3-d]pyridazin-5(6H)-one

A mixture of 4-((1S,2S)-1-(dimethylamino)-2-((6-methyl-5-oxo-5,6-dihydropyrido[2,3-d]pyridazin-8-yl)amino)propyl)benzoic acid (35 mg, 0.09 mmol), morpholine (9 mg, 0.10 mmol), HATU (42 mg, 0.11 mmol) and TEA (11 mg, 0.11 mmol) in DMF (8 mL) was stirred at room temperature for 10 h, at which point LCMS indicated that the reaction was complete. The reaction mixture was concentrated under reduced pressure, and the residue was partitioned between dichloromethane (10 mL) and water (8 mL). The separated organic phase was concentrated under reduced pressure. The crude product was purified by reverse phase chromatography (acetonitrile 4-34%/0.1% NH ₄ OH/water) to give the title compound (31.8 mg, 77% yield) as a white solid. 1H NMR (400 MHz, methanol-d4) δ 9.05-9.03 (m, 1H), 8.70-8.68 (m, 1H), 7.88-7.86 (m, 1H), 7.49 (d, J = 8.0 Hz, 2H), 7.42 (d, J = 8.0 Hz, 2H), 4.51-4.49 (m, 1H), 3.77-3.51 (m, 12H), 2.22 (s, 6H), 1.18 (d, J = 6.0 Hz, 3H). LCMS M/Z (M+H) 451. The compound was isolated as a racemic mixture of unknown relative stereochemistry: assignment based on Example 81.

Examples 83-85

The following compounds were prepared in a manner similar to Example 82. All examples in the table below were prepared using commercially available amines.

Example 86

Isopropyl 4-((1S,2S)-1-(dimethylamino)-2-((6-methyl-5-oxo-5,6-dihydropyrido[2,3-d]pyridazin-8-yl)amino)propyl)benzoate

To a solution of 4-((1S,2S)-1-(dimethylamino)-2-((6-methyl-5-oxo-5,6-dihydropyrido[2,3-d]pyridazin-8-yl)amino)propyl)benzoic acid (35 mg, 0.09 mmol) and potassium carbonate (25 mg, 0.15 mmol) in DMF (3 mL) was added 2-iodopropane (31 mg, 0.18 mmol). The reaction mixture was stirred at ambient temperature for 4 h, at which point LCMS indicated that the reaction was complete. The mixture was concentrated under reduced pressure. The residue was diluted with ethyl acetate (15 mL), washed with water (2 x 10 mL) and concentrated under reduced pressure. The residue was purified by reverse phase chromatography (acetonitrile 15-45%/0.1% NH ₄ OH/water) to give the title compound (23 mg, 59% yield) as a white solid. 1H NMR (400 MHz, methanol-d4) δ 9.05-9.04 (m, 1H), 8.69-8.66 (m, 1H), 8.03 (d, J = 8.4 Hz, 2H), 7.87-7.84 (m, 1H), 7.42 (d, J = 8.4 Hz, 2H), 5.26-5.24 (m, 1H), 4.54-4.51 (m, 1H), 3.77-3.73 (m, 4H), 2.22 (s, 6H), 1.40 (d, J = 6.4 Hz, 6H), 1.17 (d, J = 6.4 Hz, 3H). LCMS M/Z (M+H) 424. The compound was isolated as a racemic mixture of unknown relative stereochemistry: assignment based on Example 81.

Example 87

The following compound was prepared in a similar manner to Example 86 using benzyl bromide instead of 2-iodopropane.

Example 88

4-(((1S,2S)-1-(dimethylamino)-1-(6-oxo-1,6-dihydropyridin-3-yl)propan-2-yl)amino)-2-methylphthalazin-1(2H)-one

To a solution of 4-(((1S,2S)-1-(dimethylamino)-1-(6-methoxypyridin-3-yl)propan-2-yl)amino)-2-methylphthalazin-1(2H)-one (50 mg, 0.14 mol) in acetic acid (2 mL) was added 48 wt.% aqueous HBr solution (0.2 mL). The reaction mixture was heated to 80 ° C for 3 h and then quenched in ice water. The pH was adjusted to 10 with potassium carbonate and the product was extracted with ethyl acetate (repeated 3 times). The combined organic layers were dried over Na ₂ SO ₄ , filtered and concentrated to dryness under reduced pressure. The crude product was purified by flash chromatography (dichloromethane/methanol 95:5 to 70:30) to give the title compound ( ³³ mg, 69% yield). NMR (400MHz, DMSO-d6) δ11.50 (br.s, 1H), 8.26 (dd, J＝1.42, 7.77Hz, 1H), 8.05 (d, J＝ 8.10Hz,1H),7.89(dt,J＝1.53,7.66Hz,1H),7.80-7.86(m,1H),7.39(dd,J＝2.63,9.4 3H), 4.24-4.35 (m, 1H), 3.60 (s, 3H), 3.32-3.36 (m, 1H), 2.12 (s, 6H), 1.06 (d, J = 6.35 Hz, 3H). LCMS M/Z (M+H) 354. The compound was isolated as a racemic mixture of unknown relative stereochemistry: assignment based on Example 62.

Examples 89 and 90

Step 1:

1-((1-(dimethylamino)propan-2-yl)amino)-3-methylpyrido[3,4-d]pyridazin-4(3H)-one and 4-((1-(dimethylamino)propan-2-yl)amino)-2-methylpyrido[3,4-d]pyridazin-1(2H)-one

In a pyrex vial, a 1:1 mixture of 4-chloro-2-methylpyrido[3,4-d]pyridazin-1(2H)-one and 1-chloro-3-methylpyrido[3,4-d]pyridazin-4(3H)-one (Intermediates H and I, 300 mg, 1.53 mmol), RuPhos 3rd generation precatalyst (121 mg, 0.153 mmol), and sodium tert-butoxide (442 mg, 4.6 mmol) were added. The vial was sealed, and the atmosphere was evacuated and purged with _N2 (3X). To this solution was added N1,N1-dimethylpropane-1,2-diamine (395 μL, 3.07 mmol) and toluene (5 mL). The reaction mixture was heated at 100°C for 3 h. The crude reaction mixture was concentrated in vacuo, deposited onto silica gel with methanol, and purified by flash chromatography on silica gel using a 40-100% gradient of 90:10:1 dichloromethane:methanol: _NH₄OH and dichloromethane as eluent. The product fractions were concentrated in vacuo to afford a mixture of the title compounds (335 mg, 84% yield) as a dark orange oil. LCMS M/Z (M+H) 262.

Step 2:

1-((1-(Dimethylamino)propan-2-yl)amino)-3-methyl-5,6,7,8-tetrahydropyrido[3,4-d]pyridazin-4(3H)-one and 4-((1-(Dimethylamino)propan-2-yl)amino)-2-methyl-5,6,7,8-tetrahydropyrido[3,4-d]pyridazin-1(2H)-one

A pressure vessel was charged with 4-((1-(dimethylamino)propan-2-yl)amino)-2-methylpyrido[3,4-d]pyridazin-1(2H)-one, and 1-((1-(dimethylamino)propan-2-yl)amino)-3-methyl-5,6,7,8-tetrahydropyrido[3,4-d]pyridazin-4(3H)-one (335 mg, 1.28 mmol), TFA (15 mL), and platinum(IV) oxide (60 mg, 0.26 mmol). The vessel was sealed, evacuated and purged with _N2 (3X), evacuated and purged with _H2 (3X), pressurized to 50 psi with _H2 , and mixed at ambient temperature overnight. After 15 hours, the reaction was complete with no trifluoroacetamide observed. The reaction mixture was filtered through a pad of celite washed with dichloromethane, then concentrated and used directly in the following reaction. LCMS M/Z (M+H) 266.

Step 3:

6-(4-Chlorophenyl)-1-((1-(dimethylamino)propan-2-yl)amino)-3-methyl-5,6,7,8-tetrahydropyrido[3,4-d]pyridazin-4(3H)-one and 6-(4-chlorophenyl)-4-((1-(dimethylamino)propan-2-yl)amino)-2-methyl-5,6,7,8-tetrahydropyrido[3,4-d]pyridazin-1(2H)-one

In a pyrex flask, RuPhos 3rd generation precatalyst (30 mg, 0.03 mmol) and 1-((1-(dimethylamino)propan-2-yl)amino)-3-methyl-5,6,7,8-tetrahydropyrido[3,4-d]pyridazin-4(3H)-one and 4-((1-(dimethylamino)propan-2-yl)amino)-2-methyl-5,6,7,8-tetrahydropyrido[3,4-d]pyridazin-1(2H)-one (340 mg, 1.28 mmol), sodium tert-butoxide (390 mg, 4.1 mmol), and 1-chloro-4-iodobenzene (460 mg, 1.92 mmol) were added. The vial was evacuated and purged with _N2 (3X), and then toluene (5 mL) was added. The vial was evacuated and purged with _N2 (3X), and then heated at 100°C overnight. The reaction mixture was concentrated to dryness in vacuo and purified by flash chromatography on silica gel using 90:10:1 dichloromethane:methanol: _NH₄OH as eluent. Normal phase chromatography failed to separate the regioisomeric products. Regioisomers were separated using reverse phase HPLC. A 10-55% gradient of water/MeCN (0.1% TFA) over 10 min was used. Example 89 (more polar) and Example 90 were independently treated with a saturated aqueous solution of _NaHCO₃ , extracted with dichloromethane, dried _{over Na₂SO₄} , filtered, concentrated, and lyophilized to afford the two title compounds as yellow solids. Example 89 (58 mg, 12% yield): Racemic mixture ¹ H NMR (400 MHz, DMSO-d6) δ 7.29-7.24 (m, 2H), 7.05-6.99 (m, 2H), 5.38 (d, J = 6.5 Hz, 1H), 4.01 (s, 2H), 3.84-3.75 (m, 1H), 3.52-3.45 (m, 6H), 2.42-2.34 (m, 1H), 2.23-2.13 (m, 8H), 1.14 (d, J = 6.3 Hz, 3H). LCMS M/Z (M+H) 376. Example 90 (120 mg, 25% yield): Racemic mixture ¹ H NMR (400 MHz, DMSO-d6) δ 7.31-7.26 (m, 2H), 7.15-7.10 (m, 2H), 5.53 (d, J = 7.4 Hz, 1H), 3.93 (d, J = 17.1 Hz, 4H), 3.49 (s, 3H), 3.44 (d, J = 4.6 Hz, 2H), 2.61-2.55 (m, 2H), 2.43-2.35 (m, 1H), 2.19 (s, 6H), 1.18 (d, J = 6.5 Hz, 3H). LCMS M/Z (M+H) 376.

Separation of enantiomers of the final compound

The enantiomers of some of the final compounds were separated by supercritical fluid chromatography using a PIC-100SCR instrument (100 bar pressure; 40° C. column temperature; 70 mL/min flow rate; 220 nm UV detection) under the specific conditions described below.

The enantiomers of some of the final compounds were separated by supercritical fluid chromatography (SFC) using a Waters Thar SFC-80 instrument (145 bar pressure; 38° C. column temperature; UV detection at 220 nm) under the specific conditions described below.

Biological data

_IC50 measurements against inhibitors using the PCAF AlphaLisa binding assay

The His/Flag epitope-tagged PCAF _719-832 bromodomain was cloned in-house, expressed, and purified to homogeneity. PCAF bromodomain binding and inhibition of the compounds disclosed herein were assessed by monitoring the engagement of biotinylated small molecule ligands (known to bind to the PCAF bromodomain) with the target using AlphaLisa technology (Perkin-Elmer). Specifically, ProxiPlate PCAF bromodomains (final 225 nM) were combined with biotinylated small molecule ligands (final 6 nM) in 50 mM HEPES (pH 7.5), 75 mM NaCl, 1 mM TCEP, 0.01% (w/v) BSA, and 0.008% (w/v) Brij-35 in 384 wells in the presence of DMSO (final 0.2% DMSO) or a compound dilution series in DMSO. After incubation at room temperature for 15 minutes, AlphaLisa streptavidin acceptor beads and AlphaLisa anti-histidine donor beads were added to a final concentration of 12.5 μg/mL, respectively. After 90 minutes of equilibration, the plates were read on the Envision instrument and _IC50s were calculated using a four-parameter nonlinear curve fit.

PCAF _IC50 values for Examples 1-90 were determined using the general procedure described above.

While a number of embodiments have been described, these examples can be altered to provide other embodiments that utilize the compounds and methods described herein. Accordingly, the scope of the invention is defined by the appended claims rather than the specific embodiments presented by way of example.

Claims (39)

1. A compound of formula (I) or a pharmaceutically acceptable salt thereof: in: R ¹ is selected from C _1-6 alkyl and C _2-6 alkenyl; X is NH or S; R ³ is selected from hydrogen and C _1-6 alkyl; R ⁴ is selected from hydrogen, C _1-6 alkyl, carbocyclyl and heterocyclyl, wherein each C _1-6 alkyl, carbocyclyl and heterocyclyl is optionally substituted with one or more groups R ^b ; or R ³ and R ⁴ together with the atoms to which they are attached form a carbocyclyl or heterocyclyl, said carbocyclyl or heterocyclyl being optionally substituted with one or more groups R ^b ; ^R5 is hydrogen or _C1-6 alkyl; each R ^b is independently selected from C _1-6 alkyl, carbocyclyl, oxo, halogen, -C(O)-N(R ^c ) ₂ , -OR ^c , -C(O)-OR ^c , -S(O) ₂ -R ^c and -N(R ^c )-C(O)-R ^c , wherein any C _1-6 alkyl, carbocyclyl and heterocyclyl are optionally substituted with one or more halogens; Each R ^c is independently selected from the group consisting of hydrogen, C _1-6 alkyl, carbocyclyl and heterocyclyl, wherein each C _1-6 alkyl, carbocyclyl and heterocyclyl is optionally substituted with one or more carbocyclyl groups; or two R ^c together with the nitrogen to which they are attached form a heterocyclyl; Ring A is a 5-membered or 6-membered heterocyclyl or a 5-membered or 6-membered carbocyclyl, wherein the 5-membered or 6-membered heterocyclyl and the 5-membered or 6-membered carbocyclyl are optionally substituted with one or more groups independently selected from the following: C _1-6 alkyl, -OR ^d , carbocyclyl and halogen, wherein the carbocyclyl is optionally substituted with one or more groups independently selected from halogen; Each R ^d is independently C _1-6 alkyl; Each ^Re is independently selected from the following groups: hydrogen, _C1-6 alkyl, _C1-6 alkoxy and heterocyclyl, wherein each _C1-6 alkyl is optionally substituted with one or more _C1-6 alkoxy groups; or two ^Res, together with the nitrogen to which they are attached, form a heterocyclyl optionally substituted with one or more _C1-3 alkyl; or one ^Re , ^R3 and the atom to which they are attached, form a heterocyclyl optionally substituted with one or more _C1-6 alkyl; or one ^Re , ^R4 and the atom to which they are attached, form a heterocyclyl optionally substituted with one or more _C1-6 alkyl; wherein carbocyclyl is a saturated, partially unsaturated or aromatic ring system having 3 to 12 carbon atoms; heterocyclyl is a carbocyclyl in which 1, 2, 3 or 4 carbon atoms are replaced by heteroatoms selected from O, N or S, and wherein the compound of formula (I) is not 2. The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein the compound has a formula selected from Formula I(a-j): wherein Ring A is optionally substituted with one or more groups independently selected from the group consisting of C _1-6 alkyl, -OR ^d , carbocyclyl, and halogen, wherein the carbocyclyl is optionally substituted with one or more groups independently selected from the group consisting of halogen. 3. The compound of any one of claims 1-2, wherein Ring A is optionally substituted with one or more groups independently selected from the group consisting of _C1-6 alkyl, ^-ORd , phenyl, and halogen, wherein phenyl is optionally substituted with one or more groups independently selected from halogen. 4. The compound of any one of claims 1-2, wherein Ring A is optionally substituted with one or more groups independently selected from the group consisting of methoxy, 4-chlorophenyl, fluoro, and methyl. 5. The compound according to any one of claims 1 to 2, wherein R ¹ is C _1-6 alkyl. 6. The compound of any one of claims 1-2, wherein R ¹ is methyl or ethyl. 7. The compound of any one of claims 1-2, wherein X is S. 8. The compound of any one of claims 1-2, wherein X is N(H). 9. The compound of any one of claims 1-2, wherein R ³ is hydrogen or C _1-6 alkyl. 10. The compound of any one of claims 1-2, wherein ^R3 is hydrogen, methyl or ethyl. 11. The compound of any one of claims 1-2, wherein R ⁴ is selected from H, phenyl, methyl, ethyl, cyclohexyl, 4-fluorophenyl, 4-methoxyphenyl, 4-chlorophenyl, 1-methyltriazol-4-yl, 1-phenylpyrazol-4-yl, 1-methylpyrazol-4-yl, pyridin-3-yl, 4-(phenylsulfonyl)phenyl, 4-hydroxyphenyl, 4-(methoxycarbonyl)phenyl, 2-chlorophenyl, 2-fluorophenyl, 4-carboxyphenyl, 12. The compound of any one of claims 1-2, wherein ^R3 and ^R4 together with the atoms to which they are attached form a carbocyclic or heterocyclic group selected from a cyclohexane group, a cyclopentane group, a piperidine group and an indan group, said carbocyclic or heterocyclic group being optionally substituted with one or more groups ^Rb . 13. The compound of any one of claims 1-2, wherein one ^{Re is taken} together with ^R3 and the atoms to which they are attached to form an azetidine ring, said ring optionally substituted with one or more groups independently selected from the group consisting of _C1-6 alkyl. 14. The compound of any one of claims 1-2, wherein one ^Re and ^R4 , together with the atoms to which they are attached, form a heterocyclyl optionally substituted with one or more groups independently selected from the group consisting of _C1-6 alkyl. 15. The compound of any one of claims 1-2, wherein one ^Re and ^R4 together with the atoms to which they are attached form a heterocyclic group selected from a pyrrolidine group, a piperidine group and an azepane group, said heterocyclic group being optionally substituted with one or more groups independently selected from the group consisting of _C1-6 alkyl. 16. The compound of any one of claims 1-2, wherein the group: Selected from: 17. The compound of any one of claims 1-2, wherein the group: Selected from: 18. The compound of any one of claims 1-2, wherein the group: Selected from: 19. A compound or a pharmaceutically acceptable salt thereof, wherein the compound is selected from: 20. A composition comprising a compound of formula (I) according to any one of claims 1 to 19 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable adjuvant, carrier or vehicle. 21. The composition of claim 20 in combination with another therapeutic agent. 22. The composition of claim 21, wherein the additional therapeutic agent is a chemotherapeutic agent. 23. Use of a compound of formula (I) according to any one of claims 1 to 19 or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating a PCAF-mediated disorder in an animal, wherein the disorder is cancer. 24. The use of claim 23, wherein the cancer is selected from the group consisting of carcinoma and sarcoma. 25. The use of claim 23, wherein the cancer is selected from solid tumors. 26. The use of claim 23, wherein the cancer is selected from leukemia, lung cancer, breast cancer, colon cancer, lymphoma, melanoma, neuroblastoma, ovarian cancer, rhabdomyosarcoma, squamous cell carcinoma, or renal cell carcinoma. 27. Use of a compound of formula (I) or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 19 in the preparation of a medicament for a) increasing the efficacy of a cancer treatment comprising a cytotoxic agent in an animal, b) delaying or preventing the development of cancer resistance to a cytotoxic agent in an animal, or c) prolonging the duration of response to a cancer therapy in an animal. 28. The use of claim 27, further comprising administering a cytotoxic agent to the animal. 29. Use of a compound of formula (I) according to any one of claims 1 to 19 or a pharmaceutically acceptable salt thereof in combination with a cytotoxic agent in the preparation of a medicament for the treatment of cancer. 30. The method of claim 29, wherein the cytotoxic agent is selected from the group consisting of antimicrotubule agents, platinum coordination complexes, alkylating agents, antibiotics, topoisomerase II inhibitors, antimetabolites, topoisomerase I inhibitors, hormones and hormone analogs, signal transduction pathway inhibitors, non-receptor tyrosine kinase angiogenesis inhibitors, immunotherapeutic agents, pro-apoptotic agents, inhibitors of LDH-A, inhibitors of fatty acid biosynthesis, cell cycle signaling inhibitors, HDAC inhibitors, proteasome inhibitors, and inhibitors of cancer metabolism. 31. The use of claim 29, wherein the cytotoxic agent is a taxane. 32. The use according to claim 31, wherein the taxane is paclitaxel or docetaxel. 33. The use of claim 29, wherein the cytotoxic agent is a platinum agent. 34. The use of claim 29, wherein the cytotoxic agent is an antagonist of EGFR. 35 . The use according to claim 34 , wherein the EGFR antagonist is N-(3-ethynylphenyl)-6,7-bis(2-methoxyethoxy)quinazolin-4-amine or a pharmaceutically acceptable salt thereof. 36. The use of claim 29, wherein the cytotoxic agent is a RAF inhibitor. 37. The use of claim 36, wherein the RAF inhibitor is a BRAF or CRAF inhibitor. 38. The use of claim 37, wherein the RAF inhibitor is vemurafenib. 39. The use of claim 29, wherein the cytotoxic agent is a PI3K inhibitor.