Classifications

• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D417/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00
  • C07D417/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing two hetero rings
  • C07D417/12—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing two hetero rings linked by a chain containing hetero atoms as chain links
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P35/00—Antineoplastic agents
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D413/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms
  • C07D413/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing two hetero rings
  • C07D413/12—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing two hetero rings linked by a chain containing hetero atoms as chain links
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D413/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms
  • C07D413/14—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing three or more hetero rings
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D417/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00
  • C07D417/14—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing three or more hetero rings

Definitions

• the present invention relates to compounds useful as inhibitors of protein kinases.
• the invention also provides pharmaceutically acceptable compositions comprising compounds of the present invention and methods of using said compositions in the treatment of various disorders.
• Protein kinases play a critical role in this regulatory process.
• a partial non-limiting list of such kinases includes abl, ATK, bcr-abl, BIk, Brk, Btk, c-kit, c-met, c-src, CDKl, CDK2, CDK4, CDK6, cRafl, CSFlR, CSK, EGFR, ErbB2, ErbB3, ErbB4, ERK, Fak, fes, FGFRl, FGFR2, FGFR3, FGFR4, FGFR5, Fgr, FLK4, fit— 1, Fps, Frk, Fyn, Hck, IGF-IR, INS-R, Jak, KDR, Lck, Lyn, MEK, p38, PDGFR, PIK, PKC, PYK2, ros, tiei, tie 2 , TRK
• MAPK mitogen activated protein kinase
• protein kinases have been implicated as targets in central nervous system disorders (such as Alzheimer's), inflammatory disorders (such as psoriasis, arthritis), bone diseases (such as osteoporosis), atherosclerosis, restenosis, thrombosis, metabolic disorders (such as diabetes) and infectious diseases (such as viral and fungal infections).
• central nervous system disorders such as Alzheimer's
• inflammatory disorders such as psoriasis, arthritis
• bone diseases such as osteoporosis
• atherosclerosis such as restenosis
• thrombosis such as diabetes
• infectious diseases such as viral and fungal infections
• this pathway includes a cascade of kinases in which members of the Growth Factor receptor Tyrosine Kinases (such as EGF-R, PDGF-R, VEGF-R, IGFl-R, the Insulin receptor) deliver signals through phosphorylation to other kinases such as Src Tyrosine kinase, and the Raf, Mek and Erk serine/threonine kinase families.
• members of the Growth Factor receptor Tyrosine Kinases such as EGF-R, PDGF-R, VEGF-R, IGFl-R, the Insulin receptor
• Src Tyrosine kinase Src Tyrosine kinase
• Raf Raf
• Mek Mek
• Erk serine/threonine kinase families the Raf, Mek and Erk serine/threonine kinase families.
• the loss of regulation of the growth factor signalling pathway is a frequent occurrence in cancer as well as other
• kinases The signals mediated by kinases have also been shown to control growth, death and differentiation in the cell by regulating the processes of the cell cycle. Progression through the eukaryotic cell cycle is controlled by a family of kinases called cyclin dependent kinases (CDKs).
• CDKs cyclin dependent kinases
• the regulation of CDK activation is complex, but requires the association of the CDK with a member of the cyclin family of regulatory subunits. A further level of regulation occurs through both activating and inactivating phosphorylations of the CDK subunit.
• the coordinate activation and inactivation of different cyclin/CDK complexes is necessary for normal progression through the cell cycle. Both the critical Gl-S and G2-M transitions are controlled by the activation of different cyclin/CDK activities.
• both cyclin D/CDK4 and cyclin E/CDK2 are thought to mediate the onset of S-phase. Progression through S-phase requires the activity of cyclin A/CDK2 whereas the activation of cyclin A/cdc2 (CDKl) and cyclin B/cdc2 are required for the onset of metaphase. It is not surprising, therefore, that the loss of control of CDK regulation is a frequent event in hyperproliferative diseases and cancer.
• Raf protein kinases are key components of signal transduction pathways by which specific extracellular stimuli elicit precise cellular responses in mammalian cells.
• Activated cell surface receptors activate ras/rap proteins at the inner aspect of the plasma membrane which in turn recruit and activate Raf proteins.
• Activated Raf proteins phosphorylate and activate the intracellular protein kinases MEKl and MEK2.
• activated MEKs catalyze phosphorylation and activation of p42/p44 mitogen-activated protein kinase (MAPK).
• MAPK mitogen-activated protein kinase
• Raf kinases have been suggested for use in disruption of tumor cell growth and hence in the treatment of cancers, e.g., histiocytic lymphoma, lung adenocarcinoma, small cell lung cancer, and pancreatic and breast carcinoma; and also in the treatment and/or prophylaxis of disorders associated with neuronal degeneration resulting from ischemic events, including cerebral ischemia after cardiac arrest, stroke and multi-infarct dementia and also after cerebral ischemic events such as those resulting from head injury, surgery, and/or during childbirth.
• cancers e.g., histiocytic lymphoma, lung adenocarcinoma, small cell lung cancer, and pancreatic and breast carcinoma
• prophylaxis of disorders associated with neuronal degeneration resulting from ischemic events including cerebral ischemia after cardiac arrest, stroke and multi-infarct dementia and also after cerebral ischemic events such as those resulting from head injury, surgery, and/or during childbirth.
• R x , R y , R 1 , L 1 , L 2 , Cy 1 , and Cy 2 are as defined in classes and subclasses herein, and pharmaceutical compositions thereof, as described generally and in subclasses herein, which compounds are useful as inhibitors of protein kinase (e.g., Raf), and thus are useful, for example, for the treatment of Raf-mediated diseases.
• the invention provides pharmaceutical compositions comprising a compound of the invention, wherein the compound is present in an amount effective to inhibit Raf activity.
• the invention provides pharmaceutical compositions comprising a compound of the invention and optionally further comprising an additional therapeutic agent.
• the additional therapeutic agent is an agent for the treatment of cancer.
• the present invention provides methods for inhibiting kinase (e.g., Raf) activity in a patient or a biological sample, comprising administering to said patient, or contacting said biological sample with, an effective inhibitory amount of a compound of the invention.
• the present invention provides methods for treating any disorder involving Raf activity, comprising administering to a subject in need thereof a therapeutically effective amount of a compound of the invention.
• Cy 1 is an optionally substituted phenyl or 5-6 membered saturated, partially unsaturated, or aromatic ring having 1-3 heteroatoms independently selected from nitrogen, oxygen, or sulfur;
• Cy 2 is an optionally substituted 5-14 membered saturated, partially unsaturated, or aromatic monocyclic, bicyclic, or tricyclic ring having 0-4 heteroatoms, independently selected from nitrogen, oxygen, or sulfur;
• L 1 is a direct bond or an optionally substituted, straight or branched Ci_6 alkylene chain
• L 2 is a direct bond, or is an optionally substituted, straight or branched Ci_6 alkylene chain wherein 1 or 2 methylene units of L 2 are optionally and independently replaced by -O-, -S-,
• each R is independently hydrogen or an optionally substituted Ci_6 aliphatic group
• R 1 is hydrogen or an optionally substituted Ci_6 aliphatic group
• R x and R y are taken together with their intervening atoms to form:
• Compounds of this invention include those generally set forth above and described specifically herein, and are illustrated in part by the various classes, subgenera and species disclosed herein. Additionally, the present invention provides pharmaceutically acceptable derivatives of the compounds of the invention, and methods of treating a subject using these compounds, pharmaceutical compositions thereof, or either of these in combination with one or more additional therapeutic agents.
• Certain compounds of the present invention can comprise one or more asymmetric centers, and thus can exist in various isomeric forms, e.g., stereoisomers and/or diastereomers.
• compounds of the invention and pharmaceutical compositions thereof may be in the form of an individual enantiomer, diastereomer or geometric isomer, or may be in the form of a mixture of stereoisomers.
• the compounds of the invention are enantiopure compounds.
• mixtures of stereoisomers or diastereomers are provided.
• certain compounds, as described herein, may have one or more double bonds that can exist as either the Z or E isomer, unless otherwise indicated.
• the invention additionally encompasses the compounds as individual isomers substantially free of other isomers and alternatively, as mixtures of various isomers, e.g., racemic mixtures of stereoisomers.
• this invention also encompasses pharmaceutically acceptable derivatives of these compounds and compositions comprising one or more compounds.
• a particular enantiomer may, in some embodiments be provided substantially free of the corresponding enantiomer, and may also be referred to as "optically enriched.”
• “Optically-enriched,” as used herein, means that the compound is made up of a significantly greater proportion of one enantiomer. In certain embodiments the compound is made up of at least about 90% by weight of a preferred enantiomer. In other embodiments the compound is made up of at least about 95%, 98%, or 99% by weight of a preferred enantiomer.
• Preferred enantiomers may be isolated from racemic mixtures by any method known to those skilled in the art, including chiral high pressure liquid chromatography (HPLC) and the formation and crystallization of chiral salts or prepared by asymmetric syntheses. See, for example, Jacques et al., Enantiomers, Racemates and Resolutions (Wiley Interscience, New
• heteroatom means one or more of oxygen, sulfur, nitrogen, phosphorus, or silicon (including, any oxidized form of nitrogen, sulfur, phosphorus, or silicon; the quaternized form of any basic nitrogen or; a substitutable nitrogen of a heterocyclic ring, for example N (as in 3,4-dihydro-2H-pyrrolyl), NH (as in pyrrolidinyl) or NR + (as in N-substituted pyrrolidinyl)).
• a "direct bond” or “covalent bond” refers to a single, double or triple bond. In certain embodiments, a “direct bond” refers to a single bond.
• aliphatic or "aliphatic group”, as used herein, denotes a hydrocarbon moiety that may be straight-chain (i.e., unbranched), branched, or cyclic (including fused, bridging, and spiro-fused polycyclic) and may be completely saturated or may contain one or more units of unsaturation, but which is not aromatic. Unless otherwise specified, aliphatic groups contain 1-6 carbon atoms. In some embodiments, aliphatic groups contain 1-4 carbon atoms, and in yet other embodiments aliphatic groups contain 1-3 carbon atoms.
• Suitable aliphatic groups include, but are not limited to, linear or branched, alkyl, alkenyl, and alkynyl groups, and hybrids thereof such as (cycloalkyl)alkyl, (cycloalkenyl)alkyl or (cycloalkyl)alkenyl.
• Carbocyclic used alone or as part of a larger moiety, refer to a saturated or partially unsaturated cyclic aliphatic monocyclic or bicyclic ring systems, as described herein, having from 3 to 10 members, wherein the aliphatic ring system is optionally substituted as defined above and described herein.
• Cycloaliphatic groups include, without limitation, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cycloheptyl, cycloheptenyl, cyclooctyl, cyclooctenyl, and cyclooctadienyl.
• the cycloalkyl has 3-6 carbons.
• the terms “cycloaliphatic”, “carbocycle”, “carbocyclyl”, “carbocyclo”, or “carbocyclic” also include aliphatic rings that are fused to one or more aromatic or nonaromatic rings, such as decahydronaphthyl or tetrahydronaphthyl, where the radical or point of attachment is on the aliphatic ring.
• cycloalkylene refers to a bivalent cycloalkyl group.
• a cycloalkylene group is a 1,1 -cycloalkylene group (i.e., a spiro-fused
• Exemplary 1,1 -cycloalkylene groups include L — ⁇ .
• a cycloalkylene group is a 1 ,2-cycloalkylene group or a 1,3-cycloalkylene group.
• cycloalkylene groups include and .
• alkyl refers to saturated, straight- or branched-chain hydrocarbon radicals derived from an aliphatic moiety containing between one and six carbon atoms by removal of a single hydrogen atom.
• the alkyl group employed in the invention contains 1-5 carbon atoms.
• the alkyl group employed contains 1-4 carbon atoms.
• the alkyl group contains 1-3 carbon atoms.
• the alkyl group contains 1-2 carbons.
• alkyl radicals include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, iso-butyl, sec-butyl, sec-pentyl, iso-pentyl, tert-butyl, n-pentyl, neopentyl, n-hexyl, sec-hexyl, n-heptyl, n-octyl, n-decyl, n-undecyl, dodecyl, and the like.
• alkenyl denotes a monovalent group derived from a straight- or branched-chain aliphatic moiety having at least one carbon-carbon double bond by the removal of a single hydrogen atom.
• the alkenyl group employed in the invention contains 2-6 carbon atoms.
• the alkenyl group employed in the invention contains 2-5 carbon atoms.
• the alkenyl group employed in the invention contains 2-4 carbon atoms.
• the alkenyl group employed contains 2-3 carbon atoms.
• Alkenyl groups include, for example, ethenyl, propenyl, butenyl, 1- methyl-2-buten-l-yl, and the like.
• alkynyl refers to a monovalent group derived from a straight- or branched-chain aliphatic moiety having at least one carbon-carbon triple bond by the removal of a single hydrogen atom.
• the alkynyl group employed in the invention contains 2-6 carbon atoms.
• the alkynyl group employed in the invention contains 2-5 carbon atoms.
• the alkynyl group employed in the invention contains 2-4 carbon atoms.
• the alkynyl group employed contains 2-3 carbon atoms.
• Representative alkynyl groups include, but are not limited to, ethynyl, 2-propynyl (propargyl), 1-propynyl, and the like.
• aryl used alone or as part of a larger moiety as in “aralkyl”, “aralkoxy”, or “aryloxyalkyl”, refers to monocyclic and bicyclic ring systems having a total of five to 10 ring members, wherein at least one ring in the system is aromatic and wherein each ring in the system contains three to seven ring members.
• aryl may be used interchangeably with the term “aryl ring”.
• aryl refers to an aromatic ring system which includes, but not limited to, phenyl, biphenyl, naphthyl, anthracyl and the like, which may bear one or more substituents.
• aryl is a group in which an aromatic ring is fused to one or more non-aromatic rings, such as indanyl, phthalimidyl, naphthimidyl, phenantriidinyl, or tetrahydronaphthyl, and the like.
• heteroaryl and “heteroar-”, used alone or as part of a larger moiety refer to groups having 5 to 10 ring atoms, preferably 5, 6, or 9 ring atoms; having 6, 10, or 14 ⁇ electrons shared in a cyclic array; and having, in addition to carbon atoms, from one to five heteroatoms.
• heteroatom refers to nitrogen, oxygen, or sulfur, and includes any oxidized form of nitrogen or sulfur, and any quaternized form of a basic nitrogen.
• Heteroaryl groups include, without limitation, thienyl, furanyl, pyrrolyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, oxadiazolyl, thiazolyl, isothiazolyl, thiadiazolyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, indolizinyl, purinyl, naphthyridinyl, and pteridinyl.
• heteroaryl and “heteroar-”, as used herein, also include groups in which a heteroaromatic ring is fused to one or more aryl, cycloaliphatic, or heterocyclyl rings, where the radical or point of attachment is on the heteroaromatic ring.
• Nonlimiting examples include indolyl, isoindolyl, benzothienyl, benzofuranyl, dibenzofuranyl, indazolyl, benzimidazolyl, benzthiazolyl, quinolyl, isoquinolyl, cinnolinyl, phthalazinyl, quinazolinyl, quinoxalinyl, 4H-quinolizinyl, carbazolyl, acridinyl, phenazinyl, phenothiazinyl, phenoxazinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, and pyrido[2,3-b]-l,4-oxazin- 3(4 ⁇ )-one.
• heteroaryl group may be mono- or bicyclic.
• heteroaryl may be used interchangeably with the terms “heteroaryl ring”, “heteroaryl group”, or “heteroaromatic”, any of which terms include rings that are optionally substituted.
• heteroarylkyl refers to an alkyl group substituted by a heteroaryl, wherein the alkyl and heteroaryl portions independently are optionally substituted.
• heterocycle As used herein, the terms “heterocycle”, “heterocyclyl”, “heterocyclic radical”, and “heterocyclic ring” are used interchangeably and refer to a stable 4- to 7-membered monocyclic or 7-10-membered bicyclic heterocyclic moiety that is either saturated or partially unsaturated, and having, in addition to carbon atoms, one or more, preferably one to four, heteroatoms, as defined above.
• nitrogen includes a substituted nitrogen.
• the nitrogen in a saturated or partially unsaturated ring having 0-3 heteroatoms selected from oxygen, sulfur or nitrogen, the nitrogen may be N (as in 3,4-dihydro-2H-pyrrolyl), NH (as in pyrrolidinyl), or ⁇ NR (as in ⁇ Z-substituted pyrrolidinyl).
• a heterocyclic ring can be attached to its pendant group at any heteroatom or carbon atom that results in a stable structure and any of the ring atoms can be optionally substituted.
• saturated or partially unsaturated heterocyclic radicals include, without limitation, tetrahydrofuranyl, tetrahydrothienyl, pyrrolidinyl, pyrrolidonyl, piperidinyl, pyrrolinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, decahydroquinolinyl, oxazolidinyl, piperazinyl, dioxanyl, dioxolanyl, diazepinyl, oxazepinyl, thiazepinyl, morpholinyl, and quinuclidinyl.
• heterocycle refers to an alkyl group substituted by a heterocyclyl, wherein the alkyl and heterocyclyl portions independently are optionally substituted.
• partially unsaturated refers to a ring moiety that includes at least one double or triple bond between ring atoms.
• the term “partially unsaturated” is intended to encompass rings having multiple sites of unsaturation, but is not intended to include aryl or heteroaryl moieties, as herein defined.
• alkylene refers to a bivalent alkyl group.
• An "alkylene chain” is a polymethylene group, i.e., -(CH 2 ) n -, wherein n is a positive integer, preferably from 1 to 6, from 1 to 4, from 1 to 3, from 1 to 2, or from 2 to 3.
• a substituted alkylene chain is a polymethylene group in which one or more methylene hydrogen atoms are replaced with a substituent. Suitable substituents include those described below for a substituted aliphatic group.
• an alkylene chain also can be optionally replaced by a functional group.
• An alkylene chain is "replaced" by a functional group when an internal methylene unit is replaced with the functional group. Examples of suitable “interrupting functional groups” are described in the specification and claims herein.
• compounds of the invention may contain "optionally substituted” moieties.
• substituted whether preceded by the term “optionally” or not, means that one or more hydrogens of the designated moiety are replaced with a suitable substituent.
• an “optionally substituted” group may have a suitable substituent at each substitutable position of the group, and when more than one position in any given structure may be substituted with more than one substituent selected from a specified group, the substituent may be either the same or different at every position.
• Combinations of substituents envisioned under this invention are preferably those that result in the formation of stable or chemically feasible compounds.
• stable refers to compounds that are not substantially altered when subjected to conditions to allow for their production, detection, and, in certain embodiments, their recovery, purification, and use for one or more of the purposes disclosed herein.
• Suitable monovalent substituents on R 0 are independently halogen, -(CH 2 V -2 R*, -(haloR*), -(CH 2 V 2 OH, -(CH 2 V 2 OR-, -(CH 2 ) 0 2 CH(OR*) 2 ; -O(haloR'), -CN, -N 3 , -(CH 2 V 2 C(O)R*, -(CH 2 V 2 C(O)OH, -(CH 2 V 2 C(O)OR*, -(CH 2 V 2 SR*, -(CH 2 V 2 SH, -(CH 2 VzNH 2 , -(CH 2 )o- 2 NHR*, -(CH 2 ) 0 2 NR* 2 , -NO 2 , -SiR* 3 , -OSiR* 3 , -C(O)
• Suitable divalent substituents that are bound to vicinal substitutable carbons of an "optionally substituted” group include: -O(CR * 2 ) 2 3 O-, wherein each independent occurrence of R * is selected from hydrogen, C i_6 aliphatic which may be substituted as defined below, or an unsubstituted 5-6-membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur.
• Suitable substituents on the aliphatic group of R include halogen, -R*, -(haloR*), -OH, -OR*, -O(haloR'), -CN, -C(O)OH, -C(O)OR*, -NH 2 , -NHR*, -NR* 2 , or -NO 2 , wherein each R* is unsubstituted or where preceded by "halo" is substituted only with one or more halogens, and is independently Ci_ 4 aliphatic, -CH 2 Ph, -0(CH 2 )o-iPh, or a 5-6-membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur.
• Suitable substituents on a substitutable nitrogen of an "optionally substituted" group include -S(O) 2 NR 1 Z, -C(S)NR 1 Z, -C(NH)NR ⁇ , or -N(R 1 ) S(O) 2 R f ; wherein each R f is independently hydrogen, Ci_6 aliphatic which may be substituted as defined below, unsubstituted -OPh, or an unsubstituted 5-6-membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur, or, notwithstanding the definition above, two independent occurrences of taken together with their intervening atom(s) form an unsubstituted 3-12-membered saturated, partially unsaturated, or aryl mono- or bicyclic ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur.
• Suitable substituents on the aliphatic group of R 1 ⁇ are independently halogen, -R*, -(haloR*), -OH, -OR*, -O(haloR'), -CN, -C(O)OH, -C(O)OR*, -NH 2 , -NHR*, -NR* 2 , or -NO 2 , wherein each R* is unsubstituted or where preceded by "halo" is substituted only with one or more halogens, and is independently -CH 2 Ph, -O(CH 2 ) 0 iPh, or a 5-6-membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur.
• R x and R y are taken together with their intervening atoms to form: (a) a 5-membered partially unsaturated or aromatic fused ring having 0-3 heteroatoms independently selected from nitrogen, oxygen, or sulfur, or (b) a 7-membered partially unsaturated fused ring having 0-3 heteroatoms independently selected from nitrogen, oxygen, or sulfur, wherein any substitutable carbon on the ring formed by R x and R y is optionally substituted with -R 2 , oxo, -halo, -NO 2 , -CN, -OR 2 , -SR 2 , -N(R 3 ) 2 , -C(O)R 2 , -CO 2 R 2 , C(O)C(O)R 2 , -C(O)CH 2 C(O)R 2 , -S(O)R 2 , -S(O) 2 R 2 , -C(O)N(R 3 ) 2
• R x and R y are taken together with their intervening atoms to form a 5-membered partially unsaturated or aromatic fused ring having 0-3 heteroatoms independently selected from nitrogen, oxygen, or sulfur, wherein said ring is optionally substituted as defined above and described herein.
• R x and R y are taken together with their intervening atoms to form a 5-membered partially unsaturated or aromatic fused carbocyclic ring, wherein said ring is optionally substituted as defined above and described herein.
• R x and R y are taken together with their intervening atoms to form a 5-membered partially unsaturated fused carbocyclic ring, wherein said ring is optionally substituted as defined above and described herein.
• R x and R y are taken together with their intervening atoms to form a 5-membered aromatic fused carbocyclic ring, wherein said ring is optionally substituted as defined above and described herein.
• R x and R y are taken together to form a cyclopentenyl or cyclopentadienyl ring, wherein said ring is optionally substituted as defined above and described herein.
• R x and R y are taken together with their intervening atoms to form a 5-membered partially unsaturated fused ring having 1-3 heteroatoms independently selected from nitrogen, oxygen, or sulfur, wherein said ring is optionally substituted as defined above and described herein.
• R x and R y are taken together with their intervening atoms to form a 5-membered partially unsaturated fused ring having 1-3 nitrogen heteroatoms, wherein said ring is optionally substituted as defined above and described herein.
• R x and R y are taken together with their intervening atoms to form a 5- membered partially unsaturated fused ring having 1-2 nitrogen heteroatoms, wherein said ring is optionally substituted as defined above and described herein.
• R x and R y are taken together to form a imidazolidinone, oxazolidinone, or pyrrolidinone ring, wherein said ring is optionally substituted as defined above and described herein.
• R x and R y are taken together to form a imidazolidinone or pyrrolidinone ring, wherein said ring is optionally substituted as defined above and described herein.
• R x and R y are taken together with their intervening atoms to form a 5-membered aromatic fused ring having 1-3 heteroatoms independently selected from nitrogen, oxygen, or sulfur, wherein said ring is optionally substituted as defined above and described herein.
• R x and R y are taken together with their intervening atoms to form a 5-membered aromatic fused ring having 1 heteroatom independently selected from nitrogen, oxygen, or sulfur, wherein said ring is optionally substituted as defined above and described herein.
• R x and R y are taken together with their intervening atoms to form a 5-membered aromatic fused ring having 2 heteroatoms independently selected from nitrogen, oxygen, or sulfur, wherein said ring is optionally substituted as defined above and described herein.
• R x and R y are taken together with their intervening atoms to form a 5-membered aromatic fused ring having 2 nitrogen atoms, wherein said ring is optionally substituted as defined above and described herein.
• R x and R y are taken together with their intervening atoms to form a 5-membered aromatic fused ring having a nitrogen atom and a sulfur atom, wherein said ring is optionally substituted as defined above and described herein.
• R x and R y are taken together with their intervening atoms to form a 5-membered aromatic fused ring having a nitrogen atom and an oxygen atom, wherein said ring is optionally substituted as defined above and described herein.
• R x and R y are taken together with their intervening atoms to form a 5-membered aromatic fused ring having 3 heteroatoms independently selected from nitrogen, oxygen, or sulfur, wherein said ring is optionally substituted as defined above and described herein. In some embodiments, R x and R y are taken together with their intervening atoms to form a 5-membered aromatic fused ring having 3 nitrogen atoms, wherein said ring is optionally substituted as defined above and described herein.
• R x and R y are taken together to form a pyrrolo, pyrazolo, imidazolo, triazolo, thiopheno, furano, thiazolo, isothiazolo, thiadiazolo, oxazolo, isoxazolo, or oxadiaziolo fused ring, wherein said ring is optionally substituted as defined above and described herein.
• R x and R y are taken together to form a pyrrolo, pyrazolo, imidazolo, triazolo, or thiazolo fused ring, wherein said ring is optionally substituted as defined above and described herein.
• R x and R y are taken together with their intervening atoms to form a 7-membered partially unsaturated fused ring having 0-3 heteroatoms independently selected from nitrogen, oxygen, or sulfur, wherein said ring is optionally substituted as defined above and described herein.
• R x and R y are taken together with their intervening atoms to form a 7-membered partially unsaturated fused ring, wherein said ring is optionally substituted as defined above and described herein.
• R x and R y are taken together to form a cyclohepteno, cycloheptadieno, or cycloheptatrieno fused ring, wherein said ring is optionally substituted as defined above and described herein.
• R x and R y are taken together with their intervening atoms to form a 7-membered partially unsaturated fused ring having 1-3 heteroatoms independently selected from nitrogen, oxygen, or sulfur, wherein said ring is optionally substituted as defined above and described herein.
• R x and R y are taken together with their intervening atoms to form a 7-membered partially unsaturated fused ring having 1 heteroatom independently selected from nitrogen, oxygen, or sulfur, wherein said ring is optionally substituted as defined above and described herein.
• R x and R y are taken together with their intervening atoms to form a 7 membered partially unsaturated fused ring having 2 heteroatoms independently selected from nitrogen, oxygen, or sulfur, wherein said ring is optionally substituted as defined above and described herein.
• R x and R y are taken together with their intervening atoms to form a 7 membered partially unsaturated fused ring having 3 heteroatoms independently selected from nitrogen, oxygen, or sulfur, wherein said ring is optionally substituted as defined above and described herein.
• R x and R y are taken together to form a oxepino, oxepinono, thiepino, thiepinono, azepino, diazapino, azepinono, or diazepinono fused ring, wherein said ring is optionally substituted as defined above and described herein.
• R x and R y are taken together to form a azepino, azepinono, or diazepinono fused ring, wherein said ring is optionally substituted as defined above and described herein.
• any substitutable carbon on the ring formed by R x and R y is optionally substituted with hydrogen, -halo, oxo, or an optionally substituted group selected from Ci_ 6 aliphatic, a C 6 -io monocyclic or bicyclic aryl ring, or a 5-10 membered saturated, partially unsaturated, or aromatic monocyclic or bicyclic ring having 1-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur.
• any substitutable carbon on the ring formed by R x and R y is optionally substituted with hydrogen, oxo or an optionally substituted Ci_ 6 aliphatic group.
• any substitutable carbon on the ring formed by R x and R y is optionally substituted with hydrogen, oxo or a methyl group.
• any substitutable nitrogen on the ring formed by R x and R y is optionally substituted with hydrogen, an optionally substituted group selected from Ci_6 aliphatic, a C ⁇ -io monocyclic or bicyclic aryl ring, or a 5-10 membered saturated, partially unsaturated, or aromatic monocyclic or bicyclic ring having 1-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur.
• any substitutable nitrogen on the ring formed by R x and R y is optionally substituted with hydrogen or an optionally substituted Ci_6 aliphatic group.
• any substitutable nitrogen on the ring formed by R x and R y is optionally substituted with hydrogen or an optionally substituted methyl, ethyl or n- propyl group. In yet other embodiments, any substitutable nitrogen on the ring formed by R x and R y is optionally substituted with hydrogen or a methyl group.
• R 1 is hydrogen or an optionally substituted Ci_6 aliphatic group. In certain embodiments, R 1 is hydrogen. In other embodiments, R 1 is an optionally substituted Ci_ 6 aliphatic group. In certain embodiments, R 1 is an optionally substituted Ci_6 alkyl group. In some embodiments, R 1 is an optionally substituted Ci_3 alkyl group. In certain aspects, R 1 is an optionally substituted methyl or ethyl group. In certain embodiments, R 1 is an optionally substituted methyl group.
• L 1 is a direct bond or an optionally substituted, straight or branched Ci_6 alkylene chain. In some aspects, L 1 is a direct bond. In certain embodiments, L 1 is an optionally substituted, straight or branched Ci_5 alkylene chain. In some embodiments, L 1 is an optionally substituted, straight or branched alkylene chain. In other embodiments, L 1 is an optionally substituted, straight or branched Ci_ 3 alkylene chain. According to some embodiments, L 1 is an optionally substituted, straight or branched Ci_ 2 alkylene chain. [0057] In certain embodiments, L 1 is an optionally substituted, straight or branched Ci alkylene chain.
• L 1 is an optionally substituted, straight or branched C 2 alkylene chain. In other embodiments, L 1 is an optionally substituted, straight or branched C 3 alkylene chain. According to some embodiments, L 1 is an optionally substituted, straight or branched C 4 alkylene chain. In certain aspects, L 1 is an optionally substituted, straight or branched C5 alkylene chain. In other aspects, L 1 is an optionally substituted, straight or branched C 6 alkylene chain. [0058] In certain embodiments, L 1 is an optionally substituted, straight Ci_6 alkylene chain. In some embodiments, L 1 is a straight Ci_ 6 alkylene chain.
• L 1 is an optionally substituted, branched Ci_6 alkylene chain. In certain aspects, L 1 is a branched Ci_6 alkylene chain. In certain embodiments, L 1 is -CH(Ci_ 6 alkyl)-, -CH(Ci_ 5 alkyl)-, -CH(Ci 4 alkyl)-, -CH(Ci_ 3 alkyl)-, or -CH(Ci_ 2 alkyl)-. In certain embodiments, L 1 is -CH(CH 3 )-.
• Cy 1 is is an optionally substituted phenyl or an optionally substituted 5-6 membered saturated, partially unsaturated, or aromatic ring having 1-3 heteroatoms independently selected from nitrogen, oxygen, or sulfur. In some embodiments, Cy 1 is optionally substituted phenyl. In certain embodiments, Cy 1 is an optionally substituted 6 membered saturated, partially unsaturated, or aromatic ring having 1-3 heteroatoms independently selected from nitrogen, oxygen, or sulfur. In other embodiments, Cy 1 is an optionally substituted 5 -membered saturated, partially unsaturated, or aromatic ring having 1-3 heteroatoms independently selected from nitrogen, oxygen, or sulfur.
• Cy 1 is an optionally substituted 5 -membered heteroaryl ring having 2 heteroatoms independently selected from nitrogen, oxygen, or sulfur. In other embodiments, Cy 1 is an optionally substituted 5 -membered heteroaryl ring having 2 heteroatoms independently selected from nitrogen and oxygen. In some embodiments, Cy 1 is an optionally substituted 5-membered heteroaryl ring having 2 heteroatoms independently selected from nitrogen and sulfur.
• Cy 1 groups include an optionally substituted pyrrolyl, pyrazolyl, imidazolyl, triazolyl, tetrazolyl, thiophenyl, furanyl, thiazolyl, isothiazolyl, thiadiazolyl, oxazolyl, isoxazolyl, or oxadiaziolyl group.
• Cy 1 is an optionally substituted thiazolyl or isoxazolyl group.
• Cy 1 is an optionally substituted thiazolyl group.
• Cy 1 is an unsubstituted thiazolyl group.
• Cy 1 is an optionally substituted isoxazolyl group.
• Cy 1 is an unsubstituted isoxazolyl group.
• L 2 is a direct bond, or is an optionally substituted, straight or branched Ci_ 6 alkylene chain wherein 1 or 2 methylene units of L 2 are optionally and independently replaced by -O-, -S-, -N(R)-, -C(O)-, -C(O)N(R)-, -N(R)C(O)N(R)-, -N(R)C(O)-, -N(R)C(O)O-, -OC(O)N(R)-, -SO 2 -, -SO 2 N(R)-, -N(R)SO 2 -, -OC(O)-, -C(O)O-, or a 3-6 membered cycloalkylene.
• L 2 is a direct bond.
• L 2 is an optionally substituted, straight or branched Ci_6 alkylene chain wherein 1 or 2 methylene units of L 2 are replaced by -O-, -S-, -N(R)-, -C(O)-, -C(O)N(R)-, -N(R)C(O)N(R)-, -N(R)C(O)-, -N(R)C(O)O-, -OC(O)N(R)-, -SO 2 -, -SO 2 N(R)-, -N(R)SO 2 -, -OC(O)-, or -C(O)O-; wherein each R is as defined above and described herein.
• L 2 is an optionally substituted, straight or branched C 1 ⁇ t alkylene chain wherein 1 or 2 methylene units of L 2 are replaced by -0-, -S-, -N(R)-, -C(O)-, -C(O)N(R)-, -N(R)C(O)-, -SO 2 -, -SO 2 N(R)-, -N(R)SO 2 -, -OC(O)-, or -C(O)O-.
• L 2 is an optionally substituted, straight or branched Ci_ 2 alkylene chain wherein 1 methylene unit of L 2 is replaced by -0-, -S-, -N(R)-, -C(O)-, -C(O)N(R)-, -N(R)C(O)-, -SO 2 -, -SO 2 N(R)-, -N(R)SO 2 -, -OC(O)-, or -C(O)O-.
• L 2 is -0-, -S-, -N(R)-, -C(O)-, -C(O)N(R)-, -N(R)C(O)-, -SO 2 -, -SO 2 N(R)-, -N(R)SO 2 -, -OC(O)-, or -C(O)O-.
• L 2 is -C(O)N(R)-, -N(R)C(O)- -SO 2 N(R)-, -N(R)SO 2 -, -OC(O)-, or -C(O)O-.
• L 2 is -C(O)N(R)- or -N(R)C(O)-. In certain embodiments, L 2 is -C(O)N(H)- or -N(H)C(O)-. In certain embodiments, L 2 is -C(O)N(H)-.
• Cy 2 is an optionally substituted 5-14 membered saturated, partially unsaturated, or aromatic monocyclic, bicyclic, or tricyclic ring having 0-4 heteroatoms, independently selected from nitrogen, oxygen, or sulfur.
• Cy 2 is an optionally substituted 5-10 membered saturated, partially unsaturated, or aromatic monocyclic ring having 1-4 heteroatoms, independently selected from nitrogen, oxygen, or sulfur. In other embodiments, Cy 2 is an optionally substituted 5-6 membered saturated, partially unsaturated, or aromatic monocyclic ring having 1-4 heteroatoms, independently selected from nitrogen, oxygen, or sulfur.
• Cy 2 is an optionally substituted 5 -membered saturated, partially unsaturated, or aromatic monocyclic ring having 1-3 heteroatoms, independently selected from nitrogen, oxygen, or sulfur. In some embodiments, Cy 2 is an optionally substituted 5 -membered saturated, partially unsaturated, or aromatic monocyclic ring having 1-2 heteroatoms, independently selected from nitrogen, oxygen, or sulfur. In other embodiments, Cy 2 is an optionally substituted 5-membered heteroaryl ring having 1-3 heteroatoms, independently selected from nitrogen, oxygen, or sulfur. In still other embodiments, Cy 2 is an optionally substituted 5-membered heteroaryl ring having 1-2 heteroatoms, independently selected from nitrogen.
• Cy 2 groups include an optionally substituted pyrrolyl, pyrazolyl, imidazolyl, triazolyl, tetrazolyl, thiophenyl, furanyl, thiazolyl, isothiazolyl, thiadiazolyl, oxazolyl, isoxazolyl, or oxadiaziolyl group.
• Cy 2 is an optionally substituted 6-membered saturated, partially unsaturated, or aromatic monocyclic ring having 1-4 heteroatoms, independently selected from nitrogen, oxygen, or sulfur. In some embodiments, Cy 2 is an optionally substituted 6-membered saturated, partially unsaturated, or aromatic monocyclic ring having 1-2 heteroatoms, independently selected from nitrogen, oxygen, or sulfur. In other embodiments, Cy 2 is an optionally substituted 6-membered heteroaryl ring having 1-4 nitrogen atoms. In certain aspects, Cy 2 is an optionally substituted 6-membered heteroaryl ring having 1-3 nitrogen atoms. In some embodiments, Cy 2 is an optionally substituted 6-membered heteroaryl ring having 1-2 nitrogen atoms.
• Cy 2 groups include an optionally substituted pyridinyl, pyrimidinyl, pyrazinyl, pyridazinyl, triazinyl, or tetrazinyl group.
• Cy 2 is an optionally substituted pyridinyl, pyrimidinyl or pyridazinyl group.
• Cy 2 is an optionally substituted 5-10 membered saturated, partially unsaturated, or aromatic bicyclic ring having 1-4 heteroatoms, independently selected from nitrogen, oxygen, or sulfur.
• Cy 2 is an optionally substituted 5,5- fused, 5,6-fused, or 6,6-fused saturated, partially unsaturated, or aromatic bicyclic ring having 1-4 heteroatoms, independently selected from nitrogen, oxygen, or sulfur.
• Cy 2 is an optionally substituted 5,5-fused, 5,6-fused, or 6,6-fused heteroaryl ring having 1-4 heteroatoms, independently selected from nitrogen, oxygen, or sulfur.
• Cy 2 is an optionally substituted 5,5-fused, 5,6-fused, or 6,6-fused heteroaryl ring having 1-4 nitrogen atoms. In other embodiments, Cy 2 is an optionally substituted 5,6-fused heteroaryl ring having 1-4 nitrogen atoms.
• Cy 2 groups include an optionally substituted pyyrolizinyl, indolyl, quinolinyl, isoquinolinyl, benzimidazolyl, imidazopyridinyl, indazolyl, purinyl, cinnolinyl, quinazolinyl, phthalazinyl, naphthridinyl, quinoxalinyl, thianaphtheneyl, or benzofuranyl group.
• Cy 2 is an optionally substituted benzimidazolyl, imidazopyridinyl or purinyl group.
• Cy 2 is an optionally substituted 8-10 membered saturated, partially unsaturated, or aromatic monocyclic or bicyclic carbocyclic ring. In some embodiments, Cy 2 is an optionally substituted 5-10 membered saturated, partially unsaturated, or aromatic monocyclic or bicyclic carbocyclic ring. In other embodiments, Cy 2 is an optionally substituted 5-6 membered saturated, partially unsaturated, or aromatic monocyclic carbocyclic ring. In certain aspects, Cy 2 is an optionally substituted 5-membered saturated or partially unsaturated carbocyclic ring. According to one embodiment, Cy 2 is an optionally substituted 6 membered saturated, partially unsaturated, or aromatic ring. In still other embodiments, Cy 2 is an optionally substituted phenyl group.
• Cy 2 is an optionally substituted 5,5-fused-, 5,6-fused, or 6,6-fused saturated, partially unsaturated, or aromatic bicyclic ring. In some embodiments, Cy 2 is an optionally substituted 5,5-fused, 5,6-fused, or 6,6-fused aromatic bicyclic ring. In other embodiments, Cy 2 is optionally substituted naphthalenyl, indanyl or indenyl group.
• Cy 2 is optionally substituted with Ci_6 aliphatic or halogen. In some embodiments, Cy 2 is optionally substituted with Cl, F, CF3, or Ci_4 alkyl. Exemplary substituents on Cy 2 include methyl, tert-butyl, and 1-methylcyclopropyl. In other embodiments, Cy 2 is mono- or di-substituted. In certain aspects, Cy 2 is optionally substituted at the meta or the para position with any one of the above-mentioned substituents. [0071] According to one aspect, the present invention provides a compound of formula II:
• each of R 1 , R x , and R y is as defined above and described in classes and subclasses herein;
• Cy 1 is an optionally substituted 5-membered saturated, partially unsaturated, or aromatic ring having 1-3 heteroatoms independently selected from nitrogen, oxygen, or sulfur;
• Cy 2 is optionally substituted phenyl or an optionally substituted 6-membered aromatic ring having 1-3 nitrogen atoms.
• the present invention provides a compound of formula IP:
• each of R 1 , R x , and R y is as defined above and described in classes and subclasses herein; each of X, Y, and Z is independently -CH-, nitrogen, oxygen, or sulfur, wherein at least one of
• X, Y, or Z is a heteroatom and the circle depicted within the ring containing X, Y, and Z indicates that said ring is aromatic; and Cy 2 is optionally substituted phenyl or an optionally substituted 6-membered aromatic ring having 1-3 nitrogen atoms.
• each of R 1 , R x , and R y is as defined above and described in classes and subclasses herein;
• Cy 1 is an optionally substituted 5-membered saturated, partially unsaturated, or aromatic ring having 1-3 heteroatoms independently selected from nitrogen, oxygen, or sulfur; and Cy 2 is optionally substituted phenyl or an optionally substituted 6-membered aromatic ring having 1-3 nitrogen atoms.
• the present invention provides a compound of formulae II-c and II-d:
• each of R 1 , R x , and R y is as defined above and described in classes and subclasses herein; each of X, Y, and Z is independently -CH-, nitrogen, oxygen, or sulfur, wherein at least one of
• X, Y, or Z is a heteroatom and the circle depicted within the ring containing X, Y, and Z indicates that said ring is aromatic; and Cy 2 is optionally substituted phenyl or an optionally substituted 6-membered aromatic ring having 1-3 nitrogen atoms.
• the present invention provides a compound of formula III:
• each of R 1 , R x , and R y is as defined above and described in classes and subclasses herein;
• Cy 1 is an optionally substituted 5-membered saturated, partially unsaturated, or aromatic ring having 1-3 heteroatoms independently selected from nitrogen, oxygen, or sulfur; and Cy 2 is an optionally substituted 8-10 membered saturated, partially unsaturated, or aromatic bicyclic ring having 1-4 heteroatoms, independently selected from nitrogen, oxygen, or sulfur.
• the present invention provides a compound of formula III':
• each of R 1 , R x , and R y is as defined above and described in classes and subclasses herein; each of X, Y, and Z is independently -CH-, nitrogen, oxygen, or sulfur, wherein at least one of
• X, Y, or Z is a heteroatom and the circle depicted within the ring containing X, Y, and Z indicates that said ring is aromatic; and Cy 2 is optionally substituted phenyl or an optionally substituted 6-membered aromatic ring having 1-4 nitrogen atoms.
• the present invention provides a compound of formulae III-a and III-b:
• each of R 1 , R x , and R y is as defined above and described in classes and subclasses herein;
• Cy 1 is an optionally substituted 5-membered saturated, partially unsaturated, or aromatic ring having 1-3 heteroatoms independently selected from nitrogen, oxygen, or sulfur; and Cy 2 is an optionally substituted 8-10 membered saturated, partially unsaturated, or aromatic bicyclic ring having 1-4 heteroatoms, independently selected from nitrogen, oxygen, or sulfur.
• the present invention provides a compound of formulae III-c and III-d: iii-c III-d or a pharmaceutically acceptable salt thereof, wherein: each of R 1 , R x , and R y is as defined above and described in classes and subclasses herein; each of X, Y, and Z is independently -CH-, nitrogen, oxygen, or sulfur, wherein at least one of
• X, Y, or Z is a heteroatom and the circle depicted within the ring containing X, Y, and Z indicates that said ring is aromatic; and Cy 2 is optionally substituted phenyl or an optionally substituted 6-membered aromatic ring having 1-4 nitrogen atoms.
• each of R 1 , R x , and R y is as defined above and described in classes and subclasses herein;
• Cy 1 is an optionally substituted 5-6 membered saturated, partially unsaturated, or aromatic ring having 1-3 heteroatoms independently selected from nitrogen, oxygen, or sulfur; and Cy 2 is optionally substituted phenyl or an optionally substituted 6-membered aromatic ring having 1-3 nitrogen atoms.
• Yet another aspect of the present invention provides a compound of formulae IV-a and IV-b:
• each of R 1 , R x , and R y is as defined above and described in classes and subclasses herein;
• Cy 1 is an optionally substituted 5-6 membered saturated, partially unsaturated, or aromatic ring having 1-3 heteroatoms independently selected from nitrogen, oxygen, or sulfur; and Cy 2 is optionally substituted phenyl or an optionally substituted 6-membered aromatic ring having 1-3 nitrogen atoms.
• the present invention provides a compound of formula IV, IV-a, or IV-b wherein Cy 1 is a 5 -membered heteroaryl ring having 1-3 heteroatoms independently selected from nitrogen, oxygen, or sulfur.
• each of R 1 , R x , R y , L 1 , L 2 , Cy 1 , and Cy 2 is selected from those groups depicted in the Schemes and in Tables 1, 2, 3, 4, and 5, infra.
• Exemplary compounds of the present invention are set forth in the Examples in the Schemes and in Tables 1 through 5, infra.
• the present invention provides a compound selected from those set forth in Table 1, or a pharmaceutically acceptable salt thereof.
• the present invention provides a compound selected from those set forth in Table 2, or a pharmaceutically acceptable salt thereof.
• the present invention provides a compound selected from those set forth in Table 3, or a pharmaceutically acceptable salt thereof.
• the present invention provides a compound selected from those set forth in Table 4, or a pharmaceutically acceptable salt thereof.
• the present invention provides a compound selected from those set forth in Table 5, or a pharmaceutically acceptable salt thereof.
• compositions are provided.
• the present invention provides compounds that are inhibitors of protein kinases ⁇ e.g., Raf kinase), and thus the present compounds are useful for the treatment of diseases, disorders, and conditions mediated by Raf kinase.
• the present invention provides a method for treating a Raf-mediated disorder.
• Raf-mediated disorder includes diseases, disorders, and conditions mediated by Raf kinase.
• Raf-mediated disorders include melanoma, leukemia, or cancers such as colon, breast, gastric, ovarian, lung, brain, larynx, cervical, renal, lymphatic system, genitourinary tract (including bladder and prostate), stomach, bone, lymphoma, melanoma, glioma, papillary thyroid, neuroblastoma, and pancreatic cancer.
• melanoma leukemia
• cancers such as colon, breast, gastric, ovarian, lung, brain, larynx, cervical, renal, lymphatic system, genitourinary tract (including bladder and prostate), stomach, bone, lymphoma, melanoma, glioma, papillary thyroid, neuroblastoma, and pancreatic cancer.
• Raf-mediated disorders further include diseases afflicting mammals which are characterized by cellular proliferation.
• diseases include, for example, blood vessel proliferative disorders, fibrotic disorders, mesangial cell proliferative disorders, and metabolic diseases.
• Blood vessel proliferative disorders include, for example, arthritis and restenosis.
• Fibrotic disorders include, for example, hepatic cirrhosis and atherosclerosis.
• Mesangial cell proliferative disorders include, for example, glomerulonephritis, diabetic nephropathy, malignant nephrosclerosis, thrombotic microangiopathy syndromes, organ transplant rejection, and glomerulopathies.
• Metabolic disorders include, for example, psoriasis, diabetes mellitus, chronic wound healing, inflammation, and neurodegenerative diseases.
• compositions comprising any of the compounds as described herein, and optionally comprise a pharmaceutically acceptable carrier, adjuvant or vehicle. In certain embodiments, these compositions optionally further comprise one or more additional therapeutic agents.
• pharmaceutically acceptable derivatives include, but are not limited to, pharmaceutically acceptable salts, esters, salts of such esters, or any other adducts or derivatives that, upon administration to a patient in need, are capable of providing, directly or indirectly, a compound as otherwise described herein, or a metabolite or residue thereof.
• the term “pharmaceutically acceptable salt” refers to those salts that are, within the scope of sound medical judgement, suitable for use in contact with the tissues of humans or animals without undue toxicity, irritation, allergic response, or the like, and are offer with a reasonable benefit/risk ratio.
• a “pharmaceutically acceptable salt” means any at least substantially non-toxic salt or salt of an ester of a compound of this invention that, upon administration to a recipient, is capable of providing, either directly or indirectly, a compound of this invention or an inhibitorily active metabolite or residue thereof.
• the term “inhibitory metabolite or residue thereof means that a metabolite or residue thereof is also an inhibitor of a Raf kinase.
• Pharmaceutically acceptable salts are well known in the art. For example, S. M. Berge et al. describe pharmaceutically acceptable salts in detail in J. Pharmaceutical Sciences, 1977, 66, 1-19, incorporated herein by reference.
• Pharmaceutically acceptable salts of the compounds of this invention include those derived from suitable inorganic and organic acids and bases.
• Examples of pharmaceutically acceptable, nontoxic acid addition salts are salts of an amino group formed with inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid and perchloric acid or with organic acids such as acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid or malonic acid or by using other methods used in the art such as ion exchange.
• salts include adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate, gluconate, hemisulfate, heptanoate, hexanoate, hydroiodide, 2-hydroxy-ethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectinate,
• Salts derived from appropriate bases include alkali metal, alkaline earth metal, ammonium and N + (Ci_4alkyl)4 salts.
• This invention also envisions the quaternization of any basic nitrogen-containing groups of the compounds disclosed herein. Water or oil-soluble or dispersable products may be obtained by such quaternization.
• Representative alkali or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, and the like.
• Further pharmaceutically acceptable salts include, when appropriate, nontoxic ammonium, quaternary ammonium, and amine cations formed using counterions such as halide, hydroxide, carboxylate, sulfate, phosphate, nitrate, loweralkyl sulfonate and aryl sulfonate.
• the pharmaceutically acceptable compositions of the present invention additionally comprise a pharmaceutically acceptable carrier, adjuvant, or vehicle, which, as used herein, includes any and all solvents, diluents, or other liquid vehicle, dispersion or suspension aids, surface active agents, isotonic agents, thickening or emulsifying agents, preservatives, solid binders, lubricants and the like, as suited to the particular dosage form desired.
• a pharmaceutically acceptable carrier, adjuvant, or vehicle which, as used herein, includes any and all solvents, diluents, or other liquid vehicle, dispersion or suspension aids, surface active agents, isotonic agents, thickening or emulsifying agents, preservatives, solid binders, lubricants and the like, as suited to the particular dosage form desired.
• Remington's Pharmaceutical Sciences, Sixteenth Edition, E. W. Martin (Mack Publishing Co., Easton, Pa., 1980) discloses various carriers used in formulating pharmaceutically acceptable compositions
• any conventional carrier medium is incompatible with the compounds of the invention, such as by producing any undesirable biological effect or otherwise interacting in a deleterious manner with any other component(s) of the pharmaceutically acceptable composition, its use is contemplated to be within the scope of this invention.
• materials which can serve as pharmaceutically acceptable carriers include, but are not limited to, ion exchangers, alumina, aluminum stearate, lecithin, serum proteins, such as human serum albumin, buffer substances such as phosphates, glycine, sorbic acid, or 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 silica, magnesium trisilicate, polyvinyl pyrrolidone, polyacrylates, waxes, polyethylene-polyoxypropylene-block polymers, wool fat, sugars such as lactose, glucose and sucrose; starches such as corn starch and potato starch; cellulose and its derivatives such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; powdered tragacanth; malt; gelatin; talc
• provided compounds may be assayed in any of the available assays known in the art for identifying compounds having kinase inhibitory activity.
• the assay may be cellular or non-cellular, in vivo or in vitro, high- or low-throughput format, etc.
• compounds of this invention were assayed for their ability to inhibit protein kinases, more specifically Raf.
• compounds of this invention which are of particular interest include those which:
• . are inhibitors of protein kinases
• compounds of the invention are Raf kinase inhibitors. In certain exemplary embodiments, compounds of the invention are Raf inhibitors. In certain exemplary embodiments, compounds of the invention have Cell ICso values ⁇ 100 ⁇ M. In certain other embodiments, compounds of the invention have Cell ICso values ⁇ 75 ⁇ M. In certain other embodiments, compounds of the invention have Cell ICso values ⁇ 50 ⁇ M. In certain other embodiments, compounds of the invention have Cell ICso values ⁇ 25 ⁇ M. In certain other embodiments, compounds of the invention have Cell ICso values ⁇ 10 ⁇ M. In certain other embodiments, compounds of the invention have Cell ICso values ⁇ 7.5 ⁇ M.
• of the invention compounds have Cell ICso values ⁇ 5 ⁇ M. In certain other embodiments, of the invention compounds have Cell ICso values ⁇ 2.5 ⁇ M. In certain other embodiments, of the invention compounds have Cell ICso values ⁇ 1 ⁇ M. In certain other embodiments, of the invention compounds have Cell ICso values ⁇ 800 nM. In certain other embodiments, of the invention compounds have Cell ICso values ⁇ 600 nM. In certain other embodiments, inventive compounds have Cell ICso values ⁇ 500 nM. In certain other embodiments, compounds of the invention have Cell ICso values ⁇ 300 nM. In certain other embodiments, compounds of the invention have Cell ICso values ⁇ 200 nM.
• a method for the treatment or lessening the severity of an Raf- mediated disease or condition comprising administering an effective amount of a compound, or a pharmaceutically acceptable composition comprising a compound to a subject in need thereof.
• an "effective amount" of the compound or pharmaceutically acceptable composition is that amount effective for treating or lessening the severity of a Raf-mediated disease or condition.
• the compounds and compositions, according to the method of the present invention may be administered using any amount and any route of administration effective for treating or lessening the severity of a Raf-mediated disease or condition.
• dosage unit form refers to a physically discrete unit of agent appropriate for the patient to be treated. It will be understood, however, that the total daily usage of the compounds and compositions of the present invention will be decided by the attending physician within the scope of sound medical judgment.
• the specific effective dose level for any particular patient or organism will depend upon a variety of factors including the disorder being treated and the severity of the disorder; the activity of the specific compound employed; the specific composition employed; the age, body weight, general health, sex and diet of the patient; the time of administration, route of administration, and rate of excretion of the specific compound employed; the duration of the treatment; drugs used in combination or coincidental with the specific compound employed, and like factors well known in the medical arts.
• patient means an animal, preferably a mammal, and most preferably a human.
• compositions of this invention can be administered to humans and other animals orally, rectally, parenterally, intracisternally, intravaginally, intraperitoneally, topically (as by powders, ointments, or drops), bucally, as an oral or nasal spray, or the like, depending on the severity of the infection being treated.
• the compounds of the invention may be administered orally or parenterally at dosage levels of about 0.01 mg/kg to about 50 mg/kg and preferably from about 1 mg/kg to about 25 mg/kg, of subject body weight per day, one or more times a day, to obtain the desired therapeutic effect.
• Liquid dosage forms for oral administration include, but are not limited to, pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups and elixirs.
• the liquid dosage forms may contain inert diluents commonly used in the art such as, for example, water or other solvents, solubilizing agents and emulsifiers such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, dimethylformamide, oils (in particular, cottonseed, groundnut, corn, germ, olive, castor, and sesame oils), glycerol, tetrahydrofurfuryl alcohol, polyethylene glycols and fatty acid esters of sorbitan, and mixtures thereof.
• inert diluents commonly used in the art such as, for example, water or other solvents,
• the oral compositions can also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.
• adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.
• injectable preparations for example, sterile injectable aqueous or oleaginous suspensions may be formulated according to the known art using suitable dispersing or wetting agents and suspending agents.
• the sterile injectable preparation may also be a sterile injectable solution, suspension or emulsion in a nontoxic parenterally acceptable diluent or solvent, for example, as a solution in 1,3-butanediol.
• acceptable vehicles and solvents that may be employed are water, Ringer's solution, U. S.
• injectable formulations can be sterilized, for example, by filtration through a bacterial-retaining filter, or by incorporating sterilizing agents in the form of sterile solid compositions which can be dissolved or dispersed in sterile water or other sterile injectable medium prior to use.
• a compound of the present invention In order to prolong the effect of a compound of the present invention, it is often desirable to slow the absorption of the compound from subcutaneous or intramuscular injection. This may be accomplished by the use of a liquid suspension of crystalline or amorphous material with poor water solubility. The rate of absorption of the compound then depends upon its rate of dissolution that, in turn, may depend upon crystal size and crystalline form. Alternatively, delayed absorption of a parenterally administered compound form is accomplished by dissolving or suspending the compound in an oil vehicle. Injectable depot forms are made by forming microencapsule matrices of the compound in biodegradable polymers such as polylactide- polyglycolide.
• the rate of compound release can be controlled.
• biodegradable polymers include poly(orthoesters) and poly(anhydrides).
• Depot injectable formulations are also prepared by entrapping the compound in liposomes or microemulsions that are compatible with body tissues.
• compositions for rectal or vaginal administration are preferably suppositories which can be prepared by mixing the compounds of this invention with suitable non-irritating excipients or carriers such as cocoa butter, polyethylene glycol or a suppository wax which are solid at ambient temperature but liquid at body temperature and therefore melt in the rectum or vaginal cavity and release the active compound.
• suitable non-irritating excipients or carriers such as cocoa butter, polyethylene glycol or a suppository wax which are solid at ambient temperature but liquid at body temperature and therefore melt in the rectum or vaginal cavity and release the active compound.
• Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules.
• the active compound is mixed with at least one inert, pharmaceutically acceptable excipient or carrier such as sodium citrate or dicalcium phosphate and/or a) fillers or extenders such as starches, lactose, sucrose, glucose, mannitol, and silicic acid, b) binders such as, for example, carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidinone, sucrose, and acacia, c) humectants such as glycerol, d) disintegrating agents such as agar — agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates, and sodium carbonate, e) solution retarding agents such as paraffin, f) absorption accelerators such as quaternary ammonium compounds, g) wetting agents such as, for example, cety
• Solid compositions of a similar type may also be employed as fillers in soft and hard- filled gelatin capsules using such excipients as lactose or milk sugar as well as high molecular weight polyethylene glycols and the like.
• the solid dosage forms of tablets, dragees, capsules, pills, and granules can be prepared with coatings and shells such as enteric coatings and other coatings well known in the pharmaceutical formulating art. They may optionally contain opacifying agents and can also be of a composition that they release the active ingredient(s) only, or preferentially, in a certain part of the intestinal tract, optionally, in a delayed manner. Examples of embedding compositions that can be used include polymeric substances and waxes. Solid compositions of a similar type may also be employed as fillers in soft and hard-filled gelatin capsules using such excipients as lactose or milk sugar as well as high molecular weight polethylene glycols and the like.
• the active compounds can also be in micro-encapsulated form with one or more excipients as noted above.
• the solid dosage forms of tablets, dragees, capsules, pills, and granules can be prepared with coatings and shells such as enteric coatings, release controlling coatings and other coatings well known in the pharmaceutical formulating art.
• the active compound may be admixed with at least one inert diluent such as sucrose, lactose or starch.
• Such dosage forms may also comprise, as is normal practice, additional substances other than inert diluents, e.g., tableting lubricants and other tableting aids such a magnesium stearate and microcrystalline cellulose.
• the dosage forms may also comprise buffering agents. They may optionally contain opacifying agents and can also be of a composition that they release the active ingredient(s) only, or preferentially, in a certain part of the intestinal tract, optionally, in a delayed manner. Examples of embedding compositions that can be used include polymeric substances and waxes.
• Dosage forms for topical or transdermal administration of a compound of this invention include ointments, pastes, creams, lotions, gels, powders, solutions, sprays, inhalants or patches.
• the active component is admixed under sterile conditions with a pharmaceutically acceptable carrier and any needed preservatives or buffers as may be required.
• Ophthalmic formulation, ear drops, and eye drops are also contemplated as being within the scope of this invention. Additionally, the present invention contemplates the use of transdermal patches, which have the added advantage of providing controlled delivery of a compound to the body. Such dosage forms can be made by dissolving or dispensing the compound in the proper medium. Absorption enhancers can also be used to increase the flux of the compound across the skin. The rate can be controlled by either providing a rate controlling membrane or by dispersing the compound in a polymer matrix or gel.
• the compounds of the invention are useful as inhibitors of protein kinases.
• the compounds of the invention are Raf kinase inhibitors, and thus, without wishing to be bound by any particular theory, the compounds and compositions are particularly useful for treating or lessening the severity of a disease, condition, or disorder where activation of Raf kinase is implicated in the disease, condition, or disorder.
• the disease, condition, or disorder may also be referred to as a "Raf-mediated disease”.
• the present invention provides a method for treating or lessening the severity of a disease, condition, or disorder where activation of Raf kinase is implicated in the disease state.
• the activity of a compound utilized in this invention as an Raf kinase inhibitor may be assayed in vitro, in vivo, ex vivo, or in a cell line.
• In vitro assays include assays that determine inhibition of either the phosphorylation activity or ATPase activity of activated Raf. Alternate in vitro assays quantitate the ability of the inhibitor to bind to Raf. Inhibitor binding may be measured by radiolabelling the inhibitor (e.g., synthesizing the inhibitor to include a radioisotope) prior to binding, isolating the inhibitor/Raf, complex and determining the amount of radiolabel bound. Alternatively, inhibitor binding may be determined by running a competition experiment where new inhibitors are incubated with Raf bound to known radioligands.
• the term "measurably inhibit”, as used herein means a measurable change in Raf activity between a sample comprising said composition and a Raf kinase and an equivalent sample comprising Raf kinase in the absence of said composition.
• the compounds and pharmaceutically acceptable compositions of the present invention can be employed in combination therapies, that is, the compounds and pharmaceutically acceptable compositions can be administered concurrently with, prior to, or subsequent to, one or more other desired therapeutics or medical procedures.
• the particular combination of therapies (therapeutics or procedures) to employ in a combination regimen will take into account compatibility of the desired therapeutics and/or procedures and the desired therapeutic effect to be achieved.
• the therapies employed may achieve a desired effect for the same disorder (for example, compound of the invention may be administered concurrently with another agent used to treat the same disorder), or they may achieve different effects (e.g., control of any adverse effects).
• additional therapeutic agents that are normally administered to treat or prevent a particular disease, or condition, are known as "appropriate for the disease, or condition, being treated”.
• other therapies, chemotherapeutic agents, or other anti-proliferative agents may be combined with the compounds of this invention to treat proliferative diseases and cancer.
• radiotherapy e.g., gamma- radiation, neutron beam radiotherapy, electron beam radiotherapy, proton therapy, brachytherapy, and systemic radioactive isotopes
• endocrine therapy e.g., gamma- radiation, neutron beam radiotherapy, electron beam radiotherapy, proton therapy, brachytherapy, and systemic radioactive isotopes
• biologic response modifiers e
• chemotherapeutic anticancer agents examples include,but are not limited to, alkylating agents (e.g. mechlorethamine, chlorambucil, cyclophosphamide, melphalan, ifosfamide), antimetabolites (e.g., methotrexate), purine antagonists and pyrimidine antagonists (e.g.
• alkylating agents e.g. mechlorethamine, chlorambucil, cyclophosphamide, melphalan, ifosfamide
• antimetabolites e.g., methotrexate
• purine antagonists and pyrimidine antagonists e.g.
• agents the inhibitors of this invention may also be combined with include, without limitation: treatments for Alzheimer's Disease such as Aricept ® and Excelon ® ; treatments for Parkinson's Disease such as L-DOPA/carbidopa, entacapone, ropinrole, pramipexole, bromocriptine, pergolide, trihexephendyl, and amantadine; agents for treating Multiple Sclerosis (MS) such as beta interferon (e.g., Avonex ® and Rebif ® ), Copaxone ® , and mitoxantrone; treatments for asthma such as albuterol and Singulair ® ; agents for treating schizophrenia such as zyprexa, risperdal, seroquel, and haloperidol; anti-inflammatory agents such as corticosteroids, TNF blockers, IL-I RA, azathioprine, cyclophosphamide, and sulfasalazine; immuno
• Those additional agents may be administered separately from composition containing a compound of the invention, as part of a multiple dosage regimen.
• those agents may be part of a single dosage form, mixed together with a compound of this invention in a single composition. If administered as part of a multiple dosage regime, the two active agents may be submitted simultaneously, sequentially or within a period of time from one another normally within five hours from one another.
• the amount of additional therapeutic agent present in the compositions of this invention will be no more than the amount that would normally be administered in a composition comprising that therapeutic agent as the only active agent.
• the amount of additional therapeutic agent in the presently disclosed compositions will range from about 50% to 100% of the amount normally present in a composition comprising that agent as the only therapeutically active agent.
• the present invention in another aspect, includes a composition for coating an implantable device comprising a compound of the present invention as described generally above, and in classes and subclasses herein, and a carrier suitable for coating said implantable device.
• the present invention includes an implantable device coated with a composition comprising a compound of the present invention as described generally above, and in classes and subclasses herein, and a carrier suitable for coating said implantable device.
• Vascular stents for example, have been used to overcome restenosis (re-narrowing of the vessel wall after injury).
• patients using stents or other implantable devices risk clot formation or platelet activation.
• a pharmaceutically acceptable composition comprising a kinase inhibitor.
• Suitable coatings and the general preparation of coated implantable devices are described in US Patents 6,099,562; 5,886,026; and 5,304,121.
• the coatings are typically biocompatible polymeric materials such as a hydrogel polymer, polymethyldisiloxane, polycaprolactone, polyethylene glycol, polylactic acid, ethylene vinyl acetate, and mixtures thereof.
• the coatings may optionally be further covered by a suitable topcoat of fluorosilicone, polysaccarides, polyethylene glycol, phospholipids or combinations thereof to impart controlled release characteristics in the composition.
• Another aspect of the invention relates to inhibiting Raf activity in a biological sample or a patient, which method comprises administering to the patient, or contacting said biological sample with a compound of the present invention or a composition comprising said compound.
• biological sample includes, without limitation, cell cultures or extracts thereof; biopsied material obtained from a mammal or extracts thereof; and blood, saliva, urine, feces, semen, tears, or other body fluids or extracts thereof.
• Inhibition of Raf kinase activity in a biological sample is useful for a variety of purposes that are known to one of skill in the art. Examples of such purposes include, but are not limited to, blood transfusion, organ-transplantation, biological specimen storage, and biological assays.
• the present invention relates to a kit for conveniently and effectively carrying out the methods in accordance with the present invention.
• the pharmaceutical pack or kit comprises one or more containers filled with one or more of the ingredients of the pharmaceutical compositions of the invention.
• kits are especially suited for the delivery of solid oral forms such as tablets or capsules.
• Such a kit preferably includes a number of unit dosages, and may also include a card having the dosages oriented in the order of their intended use.
• a memory aid can be provided, for example in the form of numbers, letters, or other markings or with a calendar insert, designating the days in the treatment schedule in which the dosages can be administered.
• placebo dosages, or calcium dietary supplements can be included to provide a kit in which a dosage is taken every day.
• Optionally associated with such container(s) can be a notice in the form prescribed by a governmental agency regulating the manufacture, use or sale of pharmaceutical products, which notice reflects approval by the agency of manufacture, use or sale for human administration.
• the reaction was stirred at 125 0 C for three hr. After removal of excess phosphorus oxychloride at reduced pressure, ice was added to quench the reaction. The reaction was partitioned between ether and water and the aqueous layer extracted twice more with ether. The combined ether layers were washed with brine and dried over anhydrous sodium sulfate.
• the contents of the flask were cooled to ⁇ 5 0 C, and 56 mL Of NH 4 OH was added dropwise while maintaining an internal temperature ⁇ 10 0 C.
• the biphasic mixture was warmed to 35 0 C and the aqueous phase was removed.
• the organic layer was washed once more with a mixture of 24 mL OfNH 4 OH and 24 mL of water at 35 0 C.
• the aqueous phase was removed and the 16 mL of heptane was added to the organic layer.
• the organic solution was then washed with a solution of 1.15 g of EDTA in 50 mL of water at 35 0 C.
• the tartaric acid solution was stirred until all the solids dissolved at RT, and then added to the Compound D solution through a coarse filter funnel at such a rate to maintain the internal temperature of the Compound D solution at 45-53 0 C.
• the coarse filter funnel was washed with an additional 40 mL of a 3:1 2- propanol: water solution.
• the stirring of combined solutions was stopped, and the contents of the flask were held at 45 0 C for 9 hr.
• the reaction mixture was cooled to 20 0 C, and the stirring was resumed. The contents of the flask were held at 20 0 C with stirring for approximately 12 hr.
• reaction was cooled to RT and to it was added a solution of compound O.7 (10.0 g, 59.9 mmol) in DMF (20 mL) over 30 min.
• the reaction mixture was stirred at RT for an additional 5 hr, whereupon ethyl acetate (150 mL) was added.
• CC [0328] Synthesis of Compound CC.
• the synthesis of CC was accomplished following Scheme U substituting l-(2-chloropyridazin-5-yl)ethanone (Bioorg. Med. Chem. 2005, 13, 3707) for l-(2-chlorothiazol-5-yl)ethanone (U.I) and 4-t-butylaniline for 4-trifluoromethylaniline.
• Scheme UUa Compound UUa can be synthesized following Scheme M substituting 3-trifluoromethylaniline for 4-methyl-3-trifluoromethyl-phenylamine.
• 4Da 37 [0085] Synthesis of Compound 37.
• a solution of compound 4Da (20 mg, 0. 04 mmol) and N-chlorosuccinimide (40 mg, 0. 3 mmol) in acetonitrile (500 ⁇ L) was heated at 140 0 C in the microwave for 60 min.
• the reaction mixture was diluted with MeOH and purified by HPLC to afford the compound 37 (3.1 mg).
• reaction mixture was the diluted with 50 mL of EtOAc and washed 3 x water 1 x brine and then dried over Na 2 SO 4 . Removed solvent in vacuo. Took residue up in EtOAc and filtered through a plug of SiO 2 eluting with EtOAc to yield 50 mg of compound 42.
• HPES 4-(2-hydroxyethyl)-l-piperazineethanesulfonic acid
• MEK refers to mitogen activated extracellular signal-related kinase kinase
• DTT refers to dithiothreitol
• API refers to allophycocyanin
• TR-FRET refers to time resolved fluorescence energy transfer
• PBS refers to phosphate buffered saline
• PMSF refers to phenyl methyl sulfonamide
• BSA bovine serum albumin
• Inhibitors were diluted 4-fold in 100% DMSO and added to a final concentration of 10 ⁇ M to 40 pM to a solution containing 12.5 nM biotin-MEK, 0.125 nM WT Raf in 50 mM HEPES, pH 7.4, 60 mM NaCl, 3 mM MgCl 2 , 2 mM DTT, 0.25 mM MnCl 2 , 0.01% BSA, and 0.01% Tween-20 and incubated for 2 hours at room temperature. The kinase reaction was started by the addition of 50 ⁇ M ATP to a final volume of 45 ⁇ l and allowed to progress for 60 minutes.
• the reaction was stopped with 15 mM EDTA and 20 nM Streptavidin-APC, 2.5 nM Polyclonal anti p-MEKl/2 (Ser217/221), 2.5 nM Eu-labeled anti-rabbit IgG were added in Lance detection buffer and 5% Superblock in PBS for a final volume of 100 ⁇ l.
• the detection reaction was incubated for 90 minutes at room temperature and then read on an Analyst plate reader using standard TR-FRET (time resolved fluorescence resonance energy transfer) settings for Eu and APC.
• Inhibitors were diluted 4-fold in 100% DMSO and added to a final concentration of 10 ⁇ M to 40 pM to a solution containing 100 nM biotin-MEK, 0.125 nM V599E Raf in 50 mM HEPES, pH 7.4, 60 mM NaCl, 3 mM MgCl 2 , 2 mM DTT, 0.25 mM MnCl 2 , 0.01% BSA, and 0.01% Tween-20 and incubated for 20 minutes at room temperature. The kinase reaction was started by the addition of 25 ⁇ M ATP to a final volume of 45 ⁇ l and allowed to progress for 60 minutes.
• the reaction was stopped with 15 mM EDTA and 20 nM Streptavidin-APC, 2.5 nM Polyclonal anti p-MEKl/2 (Ser217/221), 2.5 nM Eu-labeled anti-rabbit IgG were added in Lance detection buffer and 5% Superblock in PBS for a final volume of 100 ⁇ l.
• the detection reaction was incubated for 90 minutes at room temperature and then read on an Analyst plate reader using standard TR-FRET (time resolved fluorescence resonance energy transfer) settings for Eu and APC.
• TR-FRET time resolved fluorescence resonance energy transfer
• Inhibitors were diluted 4-fold in 100% DMSO and added to a final concentration of 10 ⁇ M to 40 pM to a solution containing 50 nM biotin-MEK, 0.075 nM C-Raf in 50 mM HEPES, pH 7.4, 60 mM NaCl, 3 mM MgCl 2 , 2 mM DTT, 0.25 mM MnCl 2 , 0.01% BSA, and 0.01% Tween-20 and incubated for 20 minutes at room temperature. The kinase reaction was started by the addition of 10 ⁇ M ATP to a final volume of 45 ⁇ l and allowed to progress for 60 minutes.
• the reaction was stopped with 15 mM EDTA and 20 nM Streptavidin-APC, 2.5 nM Polyclonal anti p-MEKl/2 (Ser217/221), 2.5 nM Eu-labeled anti-rabbit IgG were added in Lance detection buffer and 5% Superblock in PBS for a final volume of 100 ⁇ l.
• the detection reaction was incubated for 90 minutes at room temperature and then read on an Analyst plate reader using standard TR-FRET (time resolved fluorescence resonance energy transfer) settings for Eu and APC.
• step 6 After all plates were processed (step 6), placed plates on an orbital shaker and shook at room temperature for at least 15 min.
• WM-266-4 cells were seeded at a density of 10,000 cells/well in RPMI 1640 cell culture media containing 10% FBS in a 96-well flat bottom and incubated overnight at 37 0 C.
• Inhibitors were diluted 3-fold in DMSO, added to serum free RPMI 1640 cell culture media to a final concentration range of 5 ⁇ M to 2 nM, and used to treat the previously seeded WM-266-4 cells for 1-2 hr at 37 0 C.
• lysis buffer Bio-Rad Bio-Plex Lysis Buffer, Cat # 171-304011, containing 0.4% v/v lysis buffer factor 1, 0.2% v/v lysis buffer Factor 2, and 2 mM PMSF
• Phosphorylated ERK was detected using a phospho-ERK Bioplex kit (Bio-Rad, Cat # 171-304011) per the manufacturer's instructions and detected on a Bio-Plex plate reader counting 50 beads per region.

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