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Definitions

• Spleen Tyrosine Kinase is a member of the Syk family of tyrosine kinases, and is a regulator of early B-cell development as well as mature B-cell activation, signaling, and survival.
• Syk is a non-receptor tyrosine kinase that plays critical roles in immunoreceptor- and integrin-mediated signaling in a variety of cell types, including B cells, macrophages, monocytes, mast cells, eosinophils, basophils, neutrophils, dendritic cells, T cells, natural killer cells, platelets, and osteoclasts.
• Immunoreceptors as described here include classical immunoreceptors and immunoreceptor-like molecules.
• Classical immunoreceptors include B- cell and T-cell antigen receptors as well as various immunoglobulin receptors (Fc receptors).
• Immunoreceptor-like molecules are either structurally related to immunoreceptors or participate in similar signal transduction pathways and are primarily involved in non-adaptive immune functions, including neutrophil activation, natural killer cell recognition, and osteoclast activity. Integrins are cell surface receptors that play key roles in the control of leukocyte adhesion and activation in both innate and adaptive immunity.
• B-cell signaling through BCR can lead to a wide range of biological outputs, which in turn depend on the developmental stage of the B-cell.
• the magnitude and duration of BCR signals must be precisely regulated.
• Aberrant BCR-mediated signaling can cause disregulated B-cell activation and/or the formation of pathogenic autoantibodies leading to multiple autoimmune and/or inflammatory diseases.
• Mice lacking Syk show impaired maturation of B-cells, diminished immunoglobulin production, compromised T- cell-independent immune responses and marked attenuation of the sustained calcium sign upon BCR stimulation.
• a large body of evidence supports the role of B-cells and the humoral immune system in the pathogenesis of autoimmune and/or inflammatory diseases.
• Protein-based therapeutics such as Rituxan developed to deplete B-cells represent an approach to the treatment of a number of autoimmune and inflammatory diseases.
• Auto-antibodies and their resulting immune complexes are known to play pathogenic roles in autoimmune disease and/or inflammatory disease.
• the pathogenic response to these antibodies is dependent on signaling through Fc Receptors, which is, in turn, dependent upon Syk. Because of Syk's role in B-cell activation, as well as FcR dependent signaling, inhibitors of Syk can be useful as inhibitors of B- cell mediated pathogenic activity, including autoantibody production. Therefore, inhibition of Syk enzymatic activity in cells is proposed as a treatment for autoimmune disease through its effects on autoantibody production.
• Syk also plays a key role in Fc ⁇ RI mediated mast cell and eosinophil activation.
• Syk is implicated in allergic disorders including asthma.
• Syk binds to ITAM motifs on the phosphorylated gamma chain of Fc ⁇ RI via its SH2 domains and is essential for downstream signaling.
• Syk deficiency in mice is associated with impaired IgE-mediated mast cell activation as marked by defective degranulation, leukotriene release, and cytokine secretion (e.g. TNF- alpha).
• Pharmacologic agents that inhibit Syk activity have also been shown to inhibit mast cell degranulation, leukotriene release, and cytokine secretion in cell based assays.
• Syk inhibitors have been shown to inhibit antigen- induced passive cutaneous anaphylaxsis, bronchoconstriction and bronchial edema in rats.
• Treatment with Syk antisense oligonucleotides inhibits antigen-induced infiltration of eosinophils and neutrophils in an animal model of asthma.
• Syk deficient eosinophils also show impaired activation in response to Fc ⁇ RI stimulation. Therefore, small molecule inhibitors of Syk will be useful for treatment of allergy-induced inflammatory diseases including asthma.
• the inhibition of Syk activity can be useful for the treatment of allergic disorders, autoimmune diseases and inflammatory diseases such as: SLE, rheumatoid arthritis, multiple vasculitides, idiopathic thrombocytopenic purpura (ITP), myasthenia gravis, allergic rhinitis, chronic obstructive pulmonary disease (COPD), adult respiratory distress syndrome (ARDs) and asthma.
• SLE rheumatoid arthritis
• multiple vasculitides idiopathic thrombocytopenic purpura (ITP), myasthenia gravis, allergic rhinitis, chronic obstructive pulmonary disease (COPD), adult respiratory distress syndrome (ARDs) and asthma.
• ITP idiopathic thrombocytopenic purpura
• COPD chronic obstructive pulmonary disease
• ARDs adult respiratory distress syndrome
• Syk has been reported to play an important role in ligand- independent tonic signaling through the B-cell receptor, known to be an
• W is selected from optionally substituted aryl and optionally substituted heteroaryl
• A is selected from optionally substituted alkyl, optionally substituted alkoxy, optionally substituted amino, carboxy, cyano, halo, hydrogen, hydroxy and nitro;
• X is selected from CH and N;
• Q is selected from CO and SO 2 ;
• Y is selected from hydrogen, optionally substituted alkyl; optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heteroaryl, and optionally substituted heterocycloalkyl;
• Z is selected from hydrogen, optionally substituted alkyl; optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heteroaryl, and optionally substituted heterocycloalkyl; or
• W is selected from optionally substituted aryl and optionally substituted heteroaryl
• A is selected from optionally substituted alkyl, optionally substituted alkoxy, optionally substituted amino, carboxy, cyano, halo, hydrogen, hydroxy and nitro;
• X is selected from CH and N;
• Q is selected from CO and SO 2 ;
• Y is selected from hydrogen, optionally substituted alkyl; optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heteroaryl, and optionally substituted heterocycloalkyl;
• Z is selected from hydrogen, optionally substituted alkyl; optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heteroaryl, and optionally substituted heterocycloalkyl; or
• Z taken together with Y, and the nitrogen to which they are bound, form a 5- to 13- membered nitrogen-containing ring; provided that if W is 4-(morpholino)phenyl, A is hydrogen, X is CH, Q is CO and Y is hydrogen, then Z is not hydroxyethyl; provided that if W is p-chlorophenyl or 4-(p-tolylsulfonamido)phenyl, A is hydrogen, X is CH, Q is CO and Y is hydrogen, then Z is not N,N-dimethylaminoethyl; and provided that if W is 4-(morpholinecarbonyl)phenyl, A is hydrogen, X is CH, Q is CO and Y is hydrogen, then Z is not 3-chlorobenzyl or 4-t-butylphenyl.
• compositions comprising at least one chemical entity described herein, together with at least one pharmaceutically acceptable vehicle chosen from carriers, adjuvants, and excipients.
• Also provided is a method for treating a patient having a disease chosen from cancer, autoimmune diseases, inflammatory diseases, acute inflammatory reactions, and allergic disorders comprising administering to the patient an effective amount of at least one chemical entity described herein.
• Also provided is a method for increasing sensitivity of cancer cells to chemotherapy comprising administering to a patient undergoing chemotherapy with a chemotherapeutic agent an amount of at least one chemical entity described herein, sufficient to increase the sensitivity of cancer cells to the chemotherapeutic agent.
• a method for inhibiting ATP hydrolysis comprising contacting cells expressing Syk with at least one chemical entity described herein in an amount sufficient to detectably decrease the level of ATP hydrolysis in vitro.
• a method for determining the presence of Syk in a sample comprising contacting the sample with at least one chemical entity described herein under conditions that permit detection of Syk activity, detecting a level of Syk activity in the sample, and therefrom determining the presence or absence of Syk in the sample.
• a method for inhibiting B-cell activity comprising contacting cells expressing Syk with at least one chemical entity described herein in an amount sufficient to detectably decrease B-cell activity in vitro.
• a dash (“-") that is not between two letters or symbols is used to indicate a point of attachment for a substituent. For example, -CONH 2 is attached through the carbon atom.
• “optional” or “optionally” is meant that the subsequently described event or circumstance may or may not occur, and that the description includes instances where the event or circumstance occurs and instances in which it does not.
• “optionally substituted alkyl” encompasses both “alkyl” and "substituted alkyl” as defined below.
• Alkyl encompasses straight chain and branched chain having the indicated number of carbon atoms, usually from 1 to 20 carbon atoms, for example 1 to 8 carbon atoms, such as 1 to 6 carbon atoms.
• Ci-C 6 alkyl encompasses both straight and branched chain alkyl of from 1 to 6 carbon atoms.
• alkyl groups include methyl, ethyl, propyl, isopropyl, n-butyl, sec-butyl, tert-butyl, pentyl, 2-pentyl, isopentyl, neopentyl, hexyl, 2-hexyl, 3- hexyl, 3-methylpentyl, and the like.
• Alkylene is another subset of alkyl, referring to the same residues as alkyl, but having two points of attachment. Alkylene groups will usually have from 2 to 20 carbon atoms, for example 2 to 8 carbon atoms, such as from 2 to 6 carbon atoms.
• Co alkylene indicates a covalent bond and Ci alkylene is a methylene group.
• alkyl residue having a specific number of carbons all geometric isomers having that number of carbons are intended to be encompassed; thus, for example, “butyl” is meant to include n-butyl, sec-butyl, isobutyl and t-butyl; “propyl” includes n-propyl and isopropyl.
• “Lower alkyl” refers to alkyl groups having 1 to 4 carbons.
• Alkenyl indicates an unsaturated branched or straight-chain alkyl group having at least one carbon-carbon double bond derived by the removal of one molecule of hydrogen from adjacent carbon atoms of the parent alkyl.
• the group may be in either the cis or trans configuration about the double bond(s).
• Typical alkenyl groups include, but are not limited to, ethenyl; propenyls such as prop-1-en-l-yl, prop-l-en-2-yl, prop-2-en-l-yl (allyl), prop-2-en-2-yl; butenyls such as but-1-en-l-yl, but-l-en-2-yl, 2-methyl-prop-l-en-l-yl, but-2-en-l-yl, but-2-en-en-
• an alkenyl group has from 2 to 20 carbon atoms and in other embodiments, from 2 to 6 carbon atoms.
• Alkynyl indicates an unsaturated branched or straight-chain alkyl group having at least one carbon-carbon triple bond derived by the removal of two molecules of hydrogen from adjacent carbon atoms of the parent alkyl.
• Typical alkynyl groups include, but are not limited to, ethynyl; propynyls such as prop-1-yn-l-yl, prop-2-yn-l-yl; butynyls such as but-l-yn-
• an alkynyl group has from 2 to 20 carbon atoms and in other embodiments, from 3 to 6 carbon atoms.
• Cycloalkyl indicates a saturated hydrocarbon ring group, having the specified number of carbon atoms, usually from 3 to 7 ring carbon atoms.
• Examples of cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl as well as bridged arid caged saturated ring groups such as norbornane.
• cycloalkyloxy refers to the group -O-cycloalkyl.
• alkoxy is meant an alkyl group of the indicated number of carbon atoms attached through an oxygen bridge such as, for example, methoxy, ethoxy, propoxy, isopropoxy, n-butoxy, sec-butoxy, tert-butoxy, pentoxy, 2-pentyloxy, isopentoxy, neopentoxy, hexoxy, 2- hexoxy, 3-hexoxy, 3-methylpentoxy, and the like.
• Alkoxy groups will usually have from 1 to 6 carbon atoms attached through the oxygen bridge.
• “Lower alkoxy” refers to alkoxy groups having 1 to 4 carbons.
• Acyl refers to the groups (alkyl)-C(O)-; (cycloalkyl)-C(O)-; (aryl)-C(O)-;
• a Ci-C 6 alkoxycarbonyl group is an alkoxy group having from 1 to 6 carbon atoms attached through its oxygen to a carbonyl linker.
• amino is meant the group -NH 2 .
• Aryl encompasses:
• 5- and 6-membered carbocyclic aromatic rings for example, benzene; bicyclic ring systems wherein at least one ring is carbocyclic and aromatic, for example, naphthalene, indane, and tetralin; and tricyclic ring systems wherein at least one ring is carbocyclic and aromatic, for example, fluorene.
• aryl includes 5- and 6-membered carbocyclic aromatic rings fused to a 5- to 7- membered heterocycloalkyl ring containing 1 or more heteroatoms chosen from N, O, and S.
• bicyclic ring systems wherein only one of the rings is a carbocyclic aromatic ring, the point of attachment may be at the carbocyclic aromatic ring or the heterocycloalkyl ring.
• Bivalent radicals formed from substituted benzene derivatives and having the free valences at ring atoms are named as substituted phenylene radicals.
• Bivalent radicals derived from univalent polycyclic hydrocarbon radicals whose names end in "-yl” by removal of one hydrogen atom from the carbon atom with the free valence are named by adding "-idene” to the name of the corresponding univalent radical, e.g., a naphthyl group with two points of attachment is termed naphthylidene.
• Aryl does not encompass or overlap in any way with heteroaryl, separately defined below. Hence, if one or more carbocyclic aromatic rings is fused with a heterocycloalkyl aromatic ring, the resulting ring system is heteroaryl, not aryl, as defined herein.
• aryloxy refers to the group -O-aryl.
• halo includes fluoro, chloro, bromo, and iodo
• halogen includes fluorine, chlorine, bromine, and iodine.
• Heteroaryl encompasses: 5- to 7-membered aromatic, monocyclic rings containing one or more, for example, from 1 to 4, or In some embodiments, from 1 to 3, heteroatoms chosen from N, O, and S, with the remaining ring atoms being carbon; and bicyclic heterocycloalkyl rings containing one or more, for example, from 1 to 4, or In some embodiments, from 1 to 3, heteroatoms chosen from N, O, and S, with the remaining ring atoms being carbon and wherein at least one heteroatom is present in an aromatic ring.
• heteroaryl includes a 5- to 7-membered heterocycloalkyl, aromatic ring fused to a 5- to 7-membered cycloalkyl ring.
• bicyclic heteroaryl ring systems wherein only one of the rings contains one or more heteroatoms, the point of attachment may be at the heteroaromatic ring or the cycloalkyl ring.
• the total number of S and O atoms in the heteroaryl group exceeds 1, those heteroatoms are not adjacent to one another.
• the total number of S and O atoms in the heteroaryl group is not more than 2.
• the total number of S and O atoms in the aromatic heterocycle is not more than 1.
• heteroaryl groups include, but are not limited to, (as numbered from the linkage position assigned priority 1), 2-pyridyl, 3-pyridyl, 4-pyridyl, 2,3-pyrazinyl, 3,4- pyrazinyl, 2,4-pyrimidinyl, 3,5-pyrimidinyl, 2,3-pyrazolinyl, 2,4-imidazolinyl, isoxazolinyl, oxazolinyl, thiazolinyl, thiadiazolinyl, tetrazolyl, thienyl, benzothiophenyl, furanyl, benzofuranyl, benzoimidazolinyl, indolinyl, pyridizinyl, triazolyl, quinolinyl, pyrazolyl, and 5,6,7, 8-tetrahydroisoquinoline.
• Bivalent radicals derived from univalent heteroaryl radicals whose names end in "-yl” by removal of one hydrogen atom from the atom with the free valence are named by adding "-idene" to the name of the corresponding univalent radical, e.g., a pyridyl group with two points of attachment is a pyridylidene.
• Heteroaryl does not encompass or overlap with aryl as defined above.
• Substituted heteroaryl also includes ring systems substituted with one or more oxide (-0 ) substituents, such as pyridinyl N-oxides.
• heteroaryloxy refers to the group -O-heteroaryl.
• heterocycloalkyl is meant a single aliphatic ring, usually with 3 to 7 ring atoms, containing at least 2 carbon atoms in addition to 1-3 heteroatoms independently selected from oxygen, sulfur, and nitrogen, as well as combinations comprising at least one of the foregoing heteroatoms.
• Suitable heterocycloalkyl groups include, for example (as numbered from the linkage position assigned priority 1), 2-pyrrolinyl, 2,4-imidazolidinyl, 2,3- pyrazolidinyl, 2-piperidyl, 3-piperidyl, 4-piperdyl, and 2,5-piperzinyl.
• Morpholinyl groups are also contemplated, including 2-morpholinyl and 3 -morpholinyl (numbered wherein the oxygen is assigned priority 1).
• Substituted heterocycloalkyl also includes ring systems substituted with one or more oxo moieties, such as piperidinyl N-oxide, morpholinyl-N-oxide, 1-oxo-l- thiomorpholinyl and 1,1-dioxo-l-thiomorpholinyl.
• heterocycloalkyloxy refers to the group -O-heterocylcoalkyl.
• nitro refers to the group -NO 2 .
• phosphono refers to the group -PO 3 H 2 .
• sulfanyl includes the groups: -S-( optionally substituted (Ci-
• sulfanyl includes the group Ci-C 6 alkylsulfanyl.
• sulfinyl includes the groups: -S(O)-H, -S(O)-( optionally substituted
• sulfonyl includes the groups: -S(O 2 )-H, -S(O 2 )-( optionally substituted (Ci-C 6 )alkyl), -S(O 2 )-optionally substituted aryl), -S(O 2 )-optionally substituted heteroaryl), -S(O 2 )-(optionally substituted heterocycloalkyl) ,-S(O 2 )-(optionally substituted alkoxy), -S(O 2 )-optionally substituted aryloxy), -S(O 2 )-optionally substituted heteroaryloxy),
• substituted means that any one or more hydrogens on the designated atom or group is replaced with a selection from the indicated group, provided that the designated atom's normal valence is not exceeded.
• substituted alkyl, cycloalkyl, aryl, heterocycloalkyl, and heteroaryl refer respectively to alkyl, cycloalkyl, aryl, heterocycloalkyl, and heteroaryl wherein one or more (such as up to 5, for example, up to 3) hydrogen atoms are replaced by a substituent independently chosen from:
• guanidine guanidine wherein one or more of the guanidine hydrogens are replaced with a lower-alkyl group, -NR b R c , halo, cyano, nitro, -COR b , -CO 2 R b , -CONR b R c , -OCOR b , -OCO 2 R 3 , -OCONR b R c , -NR c COR b , -NR 0 CO 2 R 3 , -NR c C0NR b R c , -CO 2 R b , -CONRV, -NR c COR b , -SOR a , -SO 2 R 3 , -SO 2 NR b R c , and
• R b is chosen from H, optionally substituted Ci-C 6 alkyl, optionally substituted aryl, and optionally substituted heteroaryl;
• R c is chosen from hydrogen and optionally substituted Ci-C 4 alkyl; or
• R b and R c and the nitrogen to which they are attached, form an optionally substituted heterocycloalkyl group; and where each optionally substituted group is unsubstituted or independently substituted with one or more, such as one, two, or three, substituents independently selected from Ci-C 4 alkyl, aryl, heteroaryl, aryl-C
• substituted acyl refers to the groups (substituted alkyl)-C(O)-;
• substituted alkyl, cycloalkyl, aryl, heteroaryl, and heterocycloalkyl refer respectively to alkyl, cycloalkyl, aryl, heteroaryl, and heterocycloalkyl wherein one or more (such as up to 5, for example, up to 3) hydrogen atoms are replaced by a substituent independently chosen from:
• guanidine guanidine wherein one or more of the guanidine hydrogens are replaced with a lower-alkyl group, -NR b R c , halo, cyano, nitro, -COR b , -CO 2 R b , -CONR 11 R 0 , -OCOR b , -OCO 2 R 3 , -OCONR b R c , -NR c COR b , -NR 0 CO 2 R 3 , -NR c CONR b R c , -CO 2 R b , -CONR 5 R 0 , -NR c C0R b , -SOR a , -SO 2 R 3 , -SO 2 NR b R c
• R b is chosen from H, optionally substituted Ci-C 6 alkyl, optionally substituted aryl, and optionally substituted heteroaryl;
• R c is chosen from hydrogen and optionally substituted Ci-C 4 alkyl; or
• R b and R c and the nitrogen to which they are attached, form an optionally substituted heterocycloalkyl group; and where each optionally substituted group is unsubstituted or independently substituted with one or more, such as one, two, or three, substituents independently selected from Ci-C 4 alkyl, aryl, heteroaryl, aryl-Ci-C 4 alkyl-, heteroaryl-Ci-C 4 alkyl-, Ci-C 4 haloalkyl-, -OCi-C 4 alkyl, -0C r C 4 alkylphenyl, -Ci-C 4 alkyl-OH, -0C r C 4 haloalkyl, halo, -OH, -NH 2 , -C J -C 4 alkyl-NH 2 , -N(Ci-C 4 alkyl)(Ci-C 4 alkyl), -NH(C-C 4 alkyl),
• substituted alkoxy refers to alkoxy wherein the alkyl constituent is substituted (i.e., -O-(substituted alkyl)) wherein “substituted alkyl” refers to alkyl wherein one or more (such as up to 5, for example, up to 3) hydrogen atoms are replaced by a substituent independently chosen from:
• guanidine guanidine wherein one or more of the guanidine hydrogens are replaced with a lower-alkyl group, -NR b R c , halo, cyano, nitro, -C0R b , -CO 2 R b , -CONR b R c , -OCOR b , -OCO 2 R 3 , -OCONR b R c , -NR c COR b , -NR c CO 2 R a , -NR c CONR b R c , -C0 2 R b , -CONR 5 R 0 , -NR c COR b , -SOR a , -SO 2 R a , -S0 2 NR
• R b is chosen from H, optionally substituted Ci-C 6 alkyl, optionally substituted aryl, and optionally substituted heteroaryl;
• R c is chosen from hydrogen and optionally substituted Ci-C 4 alkyl; or
• R b and R c and the nitrogen to which they are attached, form an optionally substituted heterocycloalkyl group; and where each optionally substituted group is unsubstituted or independently substituted with one or more, such as one, two, or three, substituents independently selected from C)-C 4 alkyl, aryl, heteroaryl, aryl-Ci-C 4 alkyl-, heteroaryl-Ci-C 4 alkyl-, Ci-C 4 haloalkyl-, -OCi-C 4 alkyl, -OCi-C 4 alkylphenyl, -C-C 4 alkyl-OH, -OCj-C 4 haloalkyl, halo, -OH, -NH 2 , -Ci-C 4 alkyl-NH 2 , -N(C-C 4 alkyl)(C r C 4 alkyl), -NH(C-C 4 alkyl),
• a substituted alkoxy group is "polyalkoxy" or -O-(optionally substituted alkylene)-(optionally substituted alkoxy), and includes groups such as -OCH 2 CH 2 OCH 3 , and residues of glycol ethers such as polyethyleneglycol, and -0(CH 2 CH 2 O) x CH 3 , where x is an integer of 2-20, such as 2-10, and for example, 2-5.
• Another substituted alkoxy group is hydroxyalkoxy or -OCH 2 (CH 2 ) y OH, where y is an integer of 1-10, such as 1-4.
• substituted alkoxycarbonyl refers to the group (substituted alkyl)-O-
• guanidine guanidine wherein one or more of the guanidine hydrogens are replaced with a lower-alkyl group, -NR b R c , halo, cyano, nitro, -COR b , -CO 2 R b , -C0NR b R c , -OCOR b , -OCO 2 R 3 , -OCONR b R c , -NR c COR b , -NR c C0 2 R a , -NR c CONR b R c , -CO 2 R b , -CONR ⁇ , -NR c COR b , -SOR a , -SO 2 R 3 , -SO 2 NR b R c ,
• R b is chosen from H, optionally substituted Ci-C 6 alkyl, optionally substituted aryl, and optionally substituted heteroaryl;
• R c is chosen from hydrogen and optionally substituted Ci-C 4 alkyl; or
• R b and R c and the nitrogen to which they are attached, form an optionally substituted heterocycloalkyl group; and where each optionally substituted group is unsubstituted or independently substituted with one or more, such as one, two, or three, substituents independently selected from Ci-C 4 alkyl, aryl, heteroaryl, aryl-Ci-C 4 alkyl-, heteroaryl-Ci-C 4 alkyl-, Ci-C 4 haloalkyl-, -OCi-C 4 alkyl, -OCi-C 4 alkylphenyl, -Ci-C 4 alkyl-OH, -OCi-C 4 haloalkyl, halo, -OH, -NH 2 , -C 1 -C 4 alkyl-NH 2 , -N(C-C 4 alkyl)(Ci-C 4 alkyl), -NH(C 1 -C 4 alkyl),
• substituted aryloxy refers to aryloxy wherein the aryl constituent is substituted (i.e., -O-(substituted aryl)) wherein “substituted aryl” refers to aryl wherein one or more (such as up to 5, for example, up to 3) hydrogen atoms are replaced by a substituent independently chosen from:
• guanidine guanidine wherein one or more of the guanidine hydrogens are replaced with a lower-alkyl group, -NR b R c , halo, cyano, nitro, -COR b , -CO 2 R b , -CONR b R c , -OCOR b , -OCO 2 R 3 , -OCONR b R c , -NR c C0R b , -NR c C0 2 R a , -NR c C0NR b R c , -CO 2 R b , -CONR 5 R 0 , -NR c COR b , -SOR a , -SO 2 R 3 , -SO 2 NR b
• R b is chosen from H, optionally substituted Ci-C 6 alkyl, optionally substituted aryl, and optionally substituted heteroaryl;
• R c is chosen from hydrogen and optionally substituted Ci-C 4 alkyl; or
• R b and R c and the nitrogen to which they are attached, form an optionally substituted heterocycloalkyl group; and where each optionally substituted group is unsubstituted or independently substituted with one or more, such as one, two, or three, substituents independently selected from Ci-C 4 alkyl, aryl, heteroaryl, aryl-Ci-C 4 alkyl-, heteroaryl-Ci-C 4 alkyl-, Ci-C 4 haloalkyl-, -OCi-C 4 alkyl, -OCi-C 4 alkylphenyl, -Ci-C 4 alkyl-OH, -OCi-C 4 haloalkyl, halo, -OH, -NH 2 , -Ci-C 4 alkyl-NH 2 , -N(Ci-C 4 alkyl)(C,-C 4 alkyl), -NH(C-C 4 alkyl), -N(C 1 -C 4 alkyl
• substituted heteroaryloxy refers to heteroaryloxy wherein the aryl constituent is substituted (i.e., -O-(substituted heteroaryl)) wherein “substituted heteroaryl” refers to aryl wherein one or more (such as up to 5, for example, up to 3) hydrogen atoms are replaced by a substituent independently chosen from:
• R b is chosen from H, optionally substituted Ci-C 6 alkyl, optionally substituted aryl, and optionally substituted heteroaryl;
• R c is chosen from hydrogen and optionally substituted Ci-C 4 alkyl; or
• R b and R c and the nitrogen to which they are attached, form an optionally substituted heterocycloalkyl group; and where each optionally substituted group is unsubstituted or independently substituted with one or more, such as one, two, or three, substituents independently selected from Ci-C 4 alkyl, aryl, heteroaryl, aryl-Ci-C 4 alkyl-, heteroaryl-Ci-C 4 alkyl-, Ci-C 4 haloalkyl-, -OCi-C 4 alkyl, -OCi-C 4 alkylphenyl, -C,-C 4 alkyl-OH, -OC, -C 4 haloalkyl, halo, -OH, -NH 2 , -Ci-C 4 alkyl-NH 2 , -N(Ci-C 4 alkyl)(Ci-C 4 alkyl), -NH(Ci-C 4 alkyl), -N(CpC 4 alkyl
• substituted cycloalkyloxy refers to cycloalkyloxy wherein the cycloalkyl constituent is substituted (i.e., -O-(substituted cycloalkyl)) wherein "substituted cycloalkyl” refers to a cycloalkyl wherein one or more hydrogen atoms are replaced by a substituent independently chosen from: -R a , -OR > b , - /0 ⁇ v(//C- ⁇ i- /C-I 2 alkyl)O- ( /e_.g., me 4t.1hy ilened ]i•oxy- ⁇ ), -S orR» b D , guanidine, guanidine wherein one or more of the guanidine hydrogens are replaced with a lower-alkyl group, -NR b R c , halo, cyano, nitro, -COR b
• R b is chosen from H, optionally substituted Ci-C 6 alkyl, optionally substituted aryl, and optionally substituted heteroaryl;
• R c is chosen from hydrogen and optionally substituted Ci-C 4 alkyl; or
• substituted heterocycloalkyloxy refers to heterocycloalkyloxy wherein the alkyl constituent is substituted (i.e., -O-(substituted heterocycloalkyl)) wherein “substituted heterocycloalkyl” refers to a heterocycloalkyloxy wherein one or more hydrogen atoms are replaced by a substituent independently chosen from:
• guanidine guanidine wherein one or more of the guanidine hydrogens are replaced with a lower-alkyl group, -NR R°, halo, cyano, nitro, -COR b , -CO 2 R b , -CONR b R c , -OCOR b , -OCO 2 R 3 , -OCONR b R c , -NR°C0R b , -NR 0 CO 2 R 3 , -NR c CONR b R c , -CO 2 R b , -CONR 15 R 0 , -NR c COR b , -SOR a , -SO 2 R 3 , -SO 2 NR b R c , and
• R b is chosen from H, optionally substituted Ci-C 6 alkyl, optionally substituted aryl, and optionally substituted heteroaryl;
• R° is chosen from hydrogen and optionally substituted Ci-C 4 alkyl; or
• R b and R c and the nitrogen to which they are attached, form an optionally substituted heterocycloalkyl group; and where each optionally substituted group is unsubstituted or independently substituted with one or more, such as one, two, or three, substituents independently selected from Cj-C 4 alkyl, aryl, heteroaryl, aryl-Ci-C 4 alkyl-, heteroaryl-Ci-C 4 alkyl-, Ci-C 4 haloalkyl-, -OCi-C 4 alkyl, -OCi-C 4 alkylphenyl, -Ci-C 4 alkyl-OH, -OCi-C 4 haloalkyl, halo, -OH, -NH 2 , -Ci-C 4 alkyl-NH 2 , -N(Ci-C 4 alkyl)(Ci-C 4 alkyl), -NH(Ci-C 4 alkyl), -N(Ci-C 4 alky
• substituted amino refers to the group -NHR d or -NR d R d where each
• R d is independently chosen from: hydroxy, optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted acyl, aminocarbonyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocycloalkyl, alkoxycarbonyl, sulfinyl and sulfonyl, provided that only one R d may be hydroxyl, and wherein substituted alkyl, cycloalkyl, aryl, heterocycloalkyl, and heteroaryl refer respectively to alkyl, cycloalkyl, aryl, heterocycloalkyl, and heteroaryl wherein one or more (such as up to 5, for example, up to 3) hydrogen atoms are replaced by a substituent independently chosen from:
• guanidine wherein one or more of the guanidine hydrogens are replaced with a lower-alkyl group, -NR b R c , halo, cyano, nitro, -COR b , -CO 2 R b , -CONR b R c , -OCOR b , -OCO 2 R 3 , -0C0NR b R c , -NR c COR b , -NR c C0 2 R a , -NR c CONR b R c , -CO 2 R b , -CONR 1 H 0 , -NR c C0R b , -SOR a , -SO 2 R a , -S0 2
• R b is chosen from H, optionally substituted Ci-C 6 alkyl, optionally substituted aryl, and optionally substituted heteroaryl;
• R c is chosen from hydrogen and optionally substituted Ci-C 4 alkyl; or
• R b and R c and the nitrogen to which they are attached, form an optionally substituted heterocycloalkyl group; and where each optionally substituted group is unsubstituted or independently substituted with one or more, such as one, two, or three, substituents independently selected from Ci-C 4 alkyl, aryl, heteroaryl, aryl-Ci-C 4 alkyl-, heteroaryl-Ci-C 4 alkyl-, Ci-C 4 haloalkyl-, -OCi-C 4 alkyl, -OC 1 -C 4 alkylphenyl, -C-C 4 alkyl-OH, -OCi-C 4 haloalkyl, halo, -OH, -NH 2 , -C-C 4 alkyl-NH 2 , -N(C 1 -C 4 alkyl)(C r C 4 alkyl), -NH(Ci-C 4 alkyl),
• substituted amino also refers to N-oxides of the groups -NHR d .
• N-oxides can be prepared by treatment of the corresponding amino group with, for example, hydrogen peroxide or m-chloroperoxybenzoic acid.
• the person skilled in the art is familiar with reaction conditions for carrying out the N-oxidation.
• Compounds described herein include, but are not limited to, their optical isomers, racemates, and other mixtures thereof. In those situations, the single enantiomers or diastereomers, i.e., optically active forms, can be obtained by asymmetric synthesis or by resolution of the racemates.
• Racemates can be accomplished, for example, by conventional methods such as crystallization in the presence of a resolving agent, or chromatography, using, for example a chiral high-pressure liquid chromatography (HPLC) column.
• HPLC high-pressure liquid chromatography
• such compounds include Z- and E- forms (or cis- and trans- forms) of compounds with carbon-carbon double bonds.
• chemical entities include all tautomeric forms of the compound. Such compounds also include crystal forms including polymorphs and clathrates.
• Chemical entities include, but are not limited to compounds described herein and all pharmaceutically acceptable forms thereof.
• compositions recited herein include pharmaceutically acceptable salts, solvates, chelates, non- covalent complexes, prodrugs, and mixtures thereof.
• the compounds described herein are in the form of pharmaceutically acceptable salts.
• the terms "chemical entity” and “chemical entities” also encompass pharmaceutically acceptable salts, solvates, chelates, non-covalent complexes, prodrugs, and mixtures.
• “Pharmaceutically acceptable salts” include, but are not limited to salts with inorganic acids, such as hydrochlorate, phosphate, diphosphate, hydrobromate, sulfate, sulfinate, nitrate, and like salts; as well as salts with an organic acid, such as malate, maleate, fumarate, tartrate, succinate, citrate, acetate, lactate, methanesulfonate, p-toluenesulfonate, 2- hydroxyethylsulfonate, benzoate, salicylate, stearate, and alkanoate such as acetate, HOOC- (CH 2 ) n -COOH where n is 0-4, and like salts.
• inorganic acids such as hydrochlorate, phosphate, diphosphate, hydrobromate, sulfate, sulfinate, nitrate, and like salts
• an organic acid such as malate, maleate, fumarate,
• pharmaceutically acceptable cations include, but are not limited to sodium, potassium, calcium, aluminum, lithium, and ammonium.
• the free base can be obtained by basifying a solution of the acid salt.
• an addition salt, particularly a pharmaceutically acceptable addition salt may be produced by dissolving the free base in a suitable organic solvent and treating the solution with an acid, in accordance with conventional procedures for preparing acid addition salts from base compounds.
• Those skilled in the art will recognize various synthetic methodologies that may be used to prepare non-toxic pharmaceutically acceptable addition salts.
• prodrugs also fall within the scope of chemical entities, for example ester or amide derivatives of the compounds described herein.
• the term "prodrugs” includes any compounds that become compounds described herein when administered to a patient, e.g., upon metabolic processing of the prodrug.
• Examples of prodrugs include, but are not limited to, acetate, formate, and benzoate and like derivatives of functional groups (such as alcohol or amine groups) in the compounds of described herein.
• solvate refers to the chemical entity formed by the interaction of a solvent and a compound. Suitable solvates are pharmaceutically acceptable solvates, such as hydrates, including monohydrates and hemi-hydrates.
• chelate refers to the chemical entity formed by the coordination of a compound to a metal ion at two (or more) points.
• non-covalent complex refers to the chemical entity formed by the interaction of a compound and another molecule wherein a covalent bond is not formed between the compound and the molecule.
• complexation can occur through van der Waals interactions, hydrogen bonding, and electrostatic interactions (also called ionic bonding).
• hydrogen bond refers to a form of association between an electronegative atom (also known as a hydrogen bond acceptor) and a hydrogen atom attached to a second, relatively electronegative atom (also known as a hydrogen bond donor).
• Suitable hydrogen bond donor and acceptors are well understood in medicinal chemistry (G. C. Pimentel and A. L. McClellan, The Hydrogen Bond, Freeman, San Francisco, 1960; R. Taylor and O.
• Hydrogen bond acceptor refers to a group comprising an oxygen or nitrogen, especially an oxygen or nitrogen that is sp -hybridized, an ether oxygen, or the oxygen of a sulfoxide or N-oxide.
• hydrogen bond donor refers to an oxygen, nitrogen, or heteroaromatic carbon that bears a hydrogen.group containing a ring nitrogen or a heteroaryl group containing a ring nitrogen.
• group As used herein the terms "group”, “radical” or “fragment” are synonymous and are intended to indicate functional groups or fragments of molecules attachable to a bond or other fragments of molecules.
• active agent is used to indicate a chemical entity which has biological activity.
• an “active agent” is a compound having pharmaceutical utility.
• an active agent may be an anti-cancer therapeutic.
• a therapeutically effective amount of a chemical entity described herein means an amount effective, when administered to a human or non-human patient, to provide a therapeutic benefit such as amelioration of symptoms, slowing of disease progression, or prevention of disease e.g., a therapeutically effective amount may be an amount sufficient to decrease the symptoms of a disease responsive to inhibition of Syk activity. In some embodiments, a therapeutically effective amount is an amount sufficient to reduce cancer symptoms, the symptoms of an allergic disorder, the symptoms of an autoimmune and/or inflammatory disease, or the symptoms of an acute inflammatory reaction.
• a therapeutically effective amount is an amount sufficient to decrease the number of detectable cancerous cells in an organism, detectably slow, or stop the growth of a cancerous tumor. In some embodiments, a therapeutically effective amount is an amount sufficient to shrink a cancerous tumor. In some circumstances a patient suffering from cancer may not present symptoms of being affected. In some embodiments, a therapeutically effective amount of a chemical entity is an amount sufficient to prevent a significant increase or significantly reduce the detectable level of cancerous cells or cancer markers in the patient's blood, serum, or tissues.
• a therapeutically effective amount may also be an amount sufficient, when administered to a patient, to detectably slow progression of the disease, or prevent the patient to whom the chemical entity is given from presenting symptoms of the allergic disorders and/or autoimmune and/or inflammatory disease, and/or acute inflammatory response.
• a therapeutically effective amount may also be an amount sufficient to produce a detectable decrease in the amount of a marker protein or cell type in the patient's blood or serum.
• a therapeutically effective amount is an amount of a chemical entity described herein sufficient to significantly decrease the activity of B-cells. In another example, in some embodiments a therapeutically effective amount is an amount of a chemical entity described herein sufficient to significantly decrease the number of B-cells. In another example, in some embodiments a therapeutically effective amount is an amount of a chemical entity described herein sufficient to decrease the level of anti- acetylcholine receptor antibody in a patient's blood with the disease myasthenia gravis. [0074] The term "inhibition" indicates a significant decrease in the baseline activity of a biological activity or process.
• “Inhibition of Syk activity” refers to a decrease in Syk activity as a direct or indirect response to the presence of at least one chemical entity described herein, relative to the activity of Syk in the absence of the at least one chemical entity.
• the decrease in activity may be due to the direct interaction of the compound with Syk, or due to the interaction of the chemical entity(ies) described herein with one or more other factors that in turn affect Syk activity.
• the presence of the chemical entity(ies) may decrease Syk activity by directly binding to the Syk, by causing (directly or indirectly) another factor to decrease Syk activity, or by (directly or indirectly) decreasing the amount of Syk present in the cell or organism.
• Inhibition of Syk activity also refers to observable inhibition of Syk activity in a standard biochemical assay for Syk activity, such as the ATP hydrolysis assay described below.
• the chemical entity described herein has an IC 50 value less than or equal to 1 micromolar. In some embodiments, the chemical entity has an IC 50 value less than or equal to less than 100 nanomolar. In some embodiments, the chemical entity has an IC 50 value less than or equal to 10 nanomolar.
• Inhibition of B-cell activity refers to a decrease in B-cell activity as a direct or indirect response to the presence of at least one chemical entity described herein, relative to the activity of B-cells in the absence of the at least one chemical entity.
• the decrease in activity may be due to the direct interaction of the compound with Syk or with one or more other factors that in turn affect B-cell activity.
• Inhibition of B-cell activity also refers to observable inhibition of CD86 expression in a standard assay such as the assay described below.
• the chemical entity described herein has an IC 50 value less than or equal to 10 micromolar.
• the chemical entity has an IC 5O value less than or equal to less than 1 micromolar.
• the chemical entity has an IC 50 value less than or equal to 500 nanomolar.
• "B cell activity" also includes activation, redistribution, reorganization, or capping of one or more various B cell membrane receptors, or membrane-bound immunoglobulins, e.g, IgM, IgG, and IgD.
• B cells also have membrane receptors for Fc portion of IgG in the form of either antigen-antibody complexes or aggregated IgG.
• B cells also carry membrane receptors for the activated components of complement, e.g., C3b, C3d, C4, and CIq. These various membrane receptors and membrane-bound immunoglobulins have membrane mobility and can undergo redistribution and capping that can initiate signal transduction.
• B cell activity also includes the synthesis or production of antibodies or immunoglobulins.
• Immunoglobulins are synthesized by the B cell series and have common structural features and structural units. Five immunoglobulin classes, i.e., IgG, IgA, IgM, IgD, and IgE, are recognized on the basis of structural differences of their heavy chains including the amino acid sequence and length of the polypeptide chain.
• Antibodies to a given antigen may be detected in all or several classes of immunoglobulins or may be restricted to a single class or subclass of immunoglobulin.
• Autoantibodies or autoimmune antibodies may likewise belong to one or several classes of immunoglobulins.
• rheumatoid factors are most often recognized as an IgM imnnunoglobulin, but can also consist of IgG or IgA.
• B cell activity also is intended to include a series of events leading to
• B cell clonal expansion from precursor B lymphocytes and differentiation into antibody-synthesizing plasma cells which takes place in conjunction with antigen-binding and with cytokine signals from other cells.
• Inhibition of B-cell proliferation refers to inhibition of proliferation of abnormal B-cells, such as cancerous B-cells, e.g. lymphoma B-cells and/ or inhibition of normal, non-diseased B-cells.
• the term "inhibition of B-cell proliferation” indicates any significant decrease in the number of B-cells, either in vitro or in vivo.
• an inhibition of B- cell proliferation in vitro would be any significant decrease in the number of B-cells in an in vitro sample contacted with at least one chemical entity described herein as compared to a matched sample not contacted with the chemical entity(ies).
• Inhibition of B-cell proliferation also refers to observable inhibition of B-cell proliferation in a standard thymidine incorporation assay for B-cell proliferation, such as the assay described herein.
• the chemical entity has an IC 50 value less than or equal to 10 micromolar. In some embodiments, the chemical entity has an IC 50 value less than or equal to less than 1 micromolar. In some embodiments, the chemical entity has an IC 50 value less than or equal to 500 nanomolar.
• An "allergy” or “allergic disorder” refers to acquired hypersensitivity to a substance (allergen). Allergic conditions include eczema, allergic rhinitis or coryza, hay fever, bronchial asthma, urticaria (hives) and food allergies, and other atopic conditions.
• Allergic conditions include eczema, allergic rhinitis or coryza, hay fever, bronchial asthma, urticaria (hives) and food allergies, and other atopic conditions.
• “Asthma” refers to a disorder of the respiratory system characterized by inflammation, narrowing of the airways and increased reactivity of the airways to inhaled agents. Asthma is frequently, although not exclusively associated with atopic or allergic symptoms.
• a "disease responsive to inhibition of Syk activity” is a disease in which inhibiting Syk kinase provides a therapeutic benefit such as an amelioration of symptoms, decrease in disease progression, prevention or delay of disease onset, or inhibition of aberrant activity of certain cell-types (monocytes, B-cells, and mast cells).
• Treatment or treating means any treatment of a disease in a patient, including: a) preventing the disease, that is, causing the clinical symptoms of the disease not to develop; b) inhibiting the disease; c) slowing or arresting the development of clinical symptoms; and/or d) relieving the disease, that is, causing the regression of clinical symptoms.
• Patient refers to an animal, such as a mammal, that has been or will be the object of treatment, observation or experiment. The methods described herein may be useful in both human therapy and veterinary applications.
• the patient is a mammal; in some embodiments the patient is human; and in some embodiments the patient is chosen from cats and dogs.
• W is selected from optionally substituted aryl and optionally substituted heteroaryl
• A is selected from optionally substituted alkyl, optionally substituted alkoxy, optionally substituted amino, carboxy, cyano, halo, hydrogen, hydroxy and nitro;
• X is selected from CH and N;
• Q is selected from CO and SO 2 ;
• Y is selected from hydrogen, optionally substituted alkyl; optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heteroaryl, and optionally substituted heterocycloalkyl;
• Z is selected from hydrogen, optionally substituted alkyl; optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heteroaryl, and optionally substituted heterocycloalkyl; or
• W is selected from optionally substituted aryl and optionally substituted heteroaryl
• A is selected from optionally substituted alkyl, optionally substituted alkoxy, optionally substituted amino, carboxy, cyano, halo, hydrogen, hydroxy and nitro;
• X is selected from CH and N;
• Q is selected from CO and SO 2 ;
• Y is selected from hydrogen, optionally substituted alkyl; optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heteroaryl, and optionally substituted heterocycloalkyl;
• Z is selected from hydrogen, optionally substituted alkyl; optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heteroaryl, and optionally substituted heterocycloalkyl; or
• Z taken together with Y, and the nitrogen to which they are bound, form a 5- to 13- membered nitrogen-containing ring; provided that if W is 4-(morpholino)phenyl, A is hydrogen, X is CH, Q is CO and Y is hydrogen, then Z is not hydroxyethyl; provided that if W is p-chlorophenyl or 4-(p-tolylsulfonamido)phenyl, A is hydrogen, X is CH, Q is CO and Y is hydrogen, then Z is not N,N-dimethylaminoethyl; and provided that if W is 4-(morpholinecarbonyl)phenyl, A is hydrogen, X is CH, Q is CO and Y is hydrogen, then Z is not 3-chlorobenzyl or 4-t-butylphenyl.
• Q is SO 2 .
• Q is C(O).
• W is optionally substituted aryl. In some embodiments, W is optionally substituted heteroaryl.
• A is selected from halo, optionally substituted lower alkyl, and hydrogen. In some embodiments, A is selected from halo, lower alkyl, and hydrogen. In some embodiments, A is hydrogen.
• X is CH. hi some embodiments, X is N.
• Y is selected from optionally substituted lower alkyl and hydrogen, hi some embodiments, Y is selected from lower alkyl and hydrogen, hi some embodiments, Y is hydrogen.
• Y and Z are taken together with the nitrogen to which they are bound to form a 5- to 13-membered nitrogen-containing ring, hi some embodiments, Y and Z are taken together with the nitrogen to which they are bound to form a 5- to 13-membered nitrogen-containing ring selected from optionally substituted piperazinyl, optionally substituted piperidinyl and optionally substituted diazepanyl.
• Z taken together with Y, and the nitrogen to which they are bound form an optionally substituted 5- to 13-membered nitrogen-containing ring which also contains an additional oxygen heteroatom.
• Z taken together with Y, and the nitrogen to which they are bound form an optionally substituted morpholinyl ring.
• Z is selected from hydrogen, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted cycloalkyl and optionally substituted heterocycloakyl. In some embodiments, Z is selected from optionally substituted phenyl, optionally substituted piperidinyl, optionally substituted benzodiazolyl, optionally substituted pyridinyl and optionally substituted cyclohexyl.
• Z is selected from hydrogen and lower alkyl that is optionally substituted with a group chosen from amino, (lower alkyl)amino, di-(lower alkyl)amino, hydroxy, and lower alkoxy. In some embodiments, Z is selected from hydrogen. [0099] In some embodiments, Z is represented by the following structure:
• R 6 , R 7 , R 8 , Rg and Rio are each independently selected from optionally substituted acyl, optionally substituted alkoxy, optionally substituted alkoxycarbonyl, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted amino, optionally substituted aminocarbonyl, optionally substituted aryl, optionally substituted aryloxy, carboxy, cyano, optionally substituted cycloalkyl, optionally substituted cycloalkyloxy, halo, optionally substituted heteroaryl, optionally substituted heteroaryloxy, optionally substituted heterocycloalkyl, optionally substituted heterocycloalkyloxy, hydrogen, hydroxy, nitro, phosphono, sulfanyl, sulfinyl, sulfonyl, and optionally substituted thiocarbonyl; or two adjacent substituents selected from R 6 , R 7 , R 8 , R 9 and R ⁇
• V is selected from C and N, wherein if V is N, then R 9 is absent. [00100] In some embodiments, Y and Z are both hydrogen.
• V is C. In some embodiments, V is N.
• R 6 , R 7 , R 8 , R 9 and Rio are each independently selected from optionally substituted Ci-C 6 alkoxy, optionally substituted Cj-C 6 alkyl, optionally substituted Ci-C 6 alkyl carbonyl, optionally substituted aminocarbonyl, optionally substituted aryl, carboxy, cyano, optionally substituted Ci-C 6 alkoxycarbonyl, halo, optionally substituted heteroaryl, hydrogen, hydroxy, and sulfonyl.
• R 6 , R 7 , R 9 and Rio are each hydrogen.
• R 8 is selected from optionally substituted Cj-C 6 alkyl, optionally substituted Ci-C 6 alkoxy, optionally substituted C)-C 6 alkoxycarbonyl, optionally substituted aminocarbonyl, carboxy, cyano, halo, optionally substituted heteroaryl, optionally substituted heterocycloalkyl, hydroxyl and sulfonyl.
• R 8 is carboxy.
• R 8 is -C(O)-NH(lower alkyl).
• R 1 , R 2 , R 3 , R 4 and R 5 are each independently selected from optionally substituted acyl, optionally substituted alkoxy, optionally substituted alkoxycarbonyl, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted amino, optionally substituted aminocarbonyl, optionally substituted aryl, optionally substituted aryloxy, carboxy, cyano, optionally substituted cycloalkyl, optionally substituted cycloalkyloxy, halo, optionally substituted heteroaryl, optionally substituted heteroaryloxy, optionally substituted heterocycloalkyl, optionally substituted heterocycloalkyloxy, hydrogen, hydroxy, nitro, phosphono, sulfanyl, sulfinyl, sulfonyl, and optionally substituted
• Ri, R 2 , R 3 , R 4 and R 5 are each independently selected from optionally substituted Ci-C 6 alkoxy, optionally substituted Ci-C 6 alkyl, optionally substituted aminocarbonyl, optionally substituted heterocycloalkyl and hydrogen. In some embodiments, Ri, R 2 and R 5 are hydrogen. In some embodiments, R 3 and R 4 are each independently selected from optionally substituted Ci-C 6 alkoxy, optionally substituted Ci-C 6 alkyl, and optionally substituted heterocycloalkyl. In some embodiments, R 3 and R 4 are both independently optionally substituted C)-C 6 alkoxy. In some embodiments, R 3 and R 4 are both methoxy.
• the chemical entities can be administered alone, as mixtures, or in combination with other active agents.
• Step 1 a solution of a compound of Formula 101 in a polar solvent such as dichloromethane is added in excess (such as about 1.1 equivalents) to a compound of Formula 102, where Ri 2 is a leaving group such as chloride.
• a tertiary amine such as ⁇ /,N-diisopropylethylamine is added to the mixture in excess (such as about 3.3 equivalents). The mixture is stirred at room temperature for about 13 h.
• the product, a compound of Formula 103 is isolated and optionally purified.
• a compound of Formula 204 (where R 13 is a halide such as Br) are taken up in an aqueous solution of base (such as IM sodium carbonate) and an inert solvent such as 1,4- dioxane.
• base such as IM sodium carbonate
• inert solvent such as 1,4- dioxane.
• the reaction mixture is sparged with nitrogen and stirred for 5 min.
• the resulting mixture is treated with about 0.2 equivalent of tetrakis(triphenylphosphine)palladium(0) and stirred to reflux for 2.5 h.
• the resulting product, a compound of Formula 205 is isolated and optionally purified.
• Step 1 a mixture of the salt of Formula 301
• a compound of Formula 101 (such as about 1.0 equivalent)
• benzotriazol-l-yloxy-tris(dimethylamino)-phosphonium hexafluorophosphate (such as about 1.0 equivalent)
• 4-methylmorpholine (such as about 3.3 equivalents) in an inert solvent such as DMF are stirred at room temperature for 18 h.
• the mixture is then stirred at 45 °C for 6 h.
• methyl 4-amino-3-methylbenzoate (such as about 1.0 equivalent) is added and the resulting mixture is stirred at room temperature for an additional 18 h.
• Step 1 a compound of Formula 205 in an aqueous mixture of MeOH is treated with HCl (such as a 2N solution (pH ⁇ 2)) and sonicated for approximately 45 min. Any solids that form are treated with a small volume of 2N HCl and
• a method of treating a patient for example, a mammal, such as a human, having a disease responsive to inhibition of Syk activity, comprising administrating to the patient having such a disease, an effective amount of at least one chemical entity described herein.
• the chemical entities described herein may also inhibit other kinases, such that disease, disease symptoms, and conditions associated with these kinases is also treated.
• Methods of treatment also include inhibiting Syk activity and/ or inhibiting B -cell activity, by inhibiting ATP binding or hydrolysis by Syk or by some other mechanism, in vivo, in a patient suffering from a disease responsive to inhibition of Syk activity, by administering an effective concentration of at least one chemical entity chosen described herein.
• An example of an effective concentration would be that concentration sufficient to inhibit Syk activity in vitro.
• An effective concentration may be ascertained experimentally, for example by assaying blood concentration of the chemical entity, or theoretically, by calculating bioavailability.
• the condition responsive to inhibition of Syk activity and/ or B-cell activity is cancer, an allergic disorder and/or an autoimmune and/or inflammatory disease, and/or an acute inflammatory reaction.
• the conditions and diseases that can be affected using chemical entities described herein include, but are not limited to: allergic disorders, including but not limited to eczema, allergic rhinitis or coryza, hay fever, bronchial asthma, urticaria (hives) and food allergies, and other atopic conditions; autoimmune and/or inflammatory diseases, including but not limited to psoriasis, Crohn's disease, irritable bowel syndrome, Sjogren's disease, tissue graft rejection, and hyperacute rejection of transplanted organs, asthma, systemic lupus erythematosus (and associated glomerulonephritis), dermatomyositis, multiple sclerosis, scleroderma ,
• Syk is a known inhibitor of apoptosis in lymphoma B-cells. Defective apoptosis contributes to the pathogenesis and drug resistance of human leukemias and lymphomas.
• a method of promoting or inducing apoptosis in cells expressing Syk comprising contacting the cell with at least one chemical entity described herein.
• a method of treating cancer, an allergic disorder and/or an autoimmune and/or inflammatory disease, and/or an acute inflammatory reaction comprises administering to a patient in need thereof an effective amount of at least one chemical entity described herein, together with a second active agent, which can be useful for treating a cancer, an allergic disorder and/or an autoimmune and/or inflammatory disease, and/or an acute inflammatory reaction.
• the second agent may be an anti-inflammatory agent.
• Treatment with the second active agent may be prior to, concomitant with, or following treatment with at least one chemical entity described herein.
• at least one chemical entity described herein is combined with another active agent in a single dosage form.
• Suitable antitumor therapeutics that may be used in combination with at least one chemical entity described herein include, but are not limited to, chemotherapeutic agents, for example mitomycin C, carboplatin, taxol, cisplatin, paclitaxel, etoposide, doxorubicin, or a combination comprising at least one of the foregoing chemotherapeutic agents. Radiotherapeutic antitumor agents may also be used, alone or in combination with chemotherapeutic agents.
• Chemical entities described herein can be useful as chemosensitizing agents, and, thus, can be useful in combination with other chemotherapeutic drugs, in particular, drugs that induce apoptosis.
• a method for increasing sensitivity of cancer cells to chemotherapy comprising administering to a patient undergoing chemotherapy a chemotherapeutic agent together with at least one chemical entity described herein in an amount sufficient to increase the sensitivity of cancer cells to the chemotherapeutic agent is also provided herein.
• chemotherapeutic drugs that can be used in combination with chemical entities described herein include topoisomerase I inhibitors (camptothesin or topotecan), topoisomerase II inhibitors (e.g. daunomycin and etoposide), alkylating agents (e.g. cyclophosphamide, melphalan and BCNU), tubulin directed agents (e.g. taxol and vinblastine), and biological agents (e.g. antibodies such as anti CD20 antibody, IDEC 8, immunotoxins, and cytokines).
• topoisomerase I inhibitors camptothesin or topotecan
• topoisomerase II inhibitors e.g. daunomycin and etoposide
• alkylating agents e.g. cyclophosphamide, melphalan and BCNU
• tubulin directed agents e.g. taxol and vinblastine
• biological agents e.g. antibodies such as anti CD20 antibody, IDEC 8, immunotoxins, and cyto
• the chemical entities described herein are used in combination with Rituxan® (Rituximab) or other agents that work by selectively depleting CD20+ B-cells.
• Antiinflammatory agents include but are not limited to NSAIDs, non-specific and COX- 2 specific cyclooxgenase enzyme inhibitors, gold compounds, corticosteroids, methotrexate, tumor necrosis factor receptor (TNF) receptors antagonists, immunosuppressants and methotrexate.
• NSAIDs include, but are not limited to ibuprofen, flurbiprofen, naproxen and naproxen sodium, diclofenac, combinations of diclofenac sodium and misoprostol, sulindac, oxaprozin, diflunisal, piroxicam, indomethacin, etodolac, fenoprofen calcium, ketoprofen, sodium nabumetone, sulfasalazine, tolmetin sodium, and hydroxychloroquine.
• NSAIDs also include COX-2 specific inhibitors (i.e., a compound that inhibits COX-2 with an IC 50 that is at least 50-fold lower than the IC 50 for COX-I) such as celecoxib, valdecoxib, lumiracoxib, etoricoxib and/or rofecoxib.
• COX-2 specific inhibitors i.e., a compound that inhibits COX-2 with an IC 50 that is at least 50-fold lower than the IC 50 for COX-I
• celecoxib valdecoxib
• lumiracoxib etoricoxib
• etoricoxib etoricoxib
• rofecoxib rofecoxib
• the anti-inflammatory agent is a salicylate.
• Salicylates include but are not limited to acetylsalicylic acid or aspirin, sodium salicylate, and choline and magnesium salicylates.
• the anti-inflammatory agent may also be a corticosteroid.
• the corticosteroid may be chosen from cortisone, dexamethasone, methylprednisolone, prednisolone, prednisolone sodium phosphate, and prednisone.
• the anti-inflammatory therapeutic agent is a gold compound such as gold sodium thiomalate or auranofin.
• the anti-inflammatory agent is a metabolic inhibitor such as a dihydrofolate reductase inhibitor, such as methotrexate or a dihydroorotate dehydrogenase inhibitor, such as leflunomide.
• a metabolic inhibitor such as a dihydrofolate reductase inhibitor, such as methotrexate or a dihydroorotate dehydrogenase inhibitor, such as leflunomide.
• At least one anti-inflammatory compound is an anti-C5 monoclonal antibody (such as eculizumab or pexelizumab), a TNF antagonist, such as entanercept, or infliximab, which is an anti-TNF alpha monoclonal antibody are used.
• an anti-C5 monoclonal antibody such as eculizumab or pexelizumab
• a TNF antagonist such as entanercept, or infliximab, which is an anti-TNF alpha monoclonal antibody
• At least one active agent is an immunosuppressant compound such as methotrexate, leflunomide, cyclosporine, tacrolimus, azathioprine, or mycophenolate mofetil are used.
• Dosage levels of the order for example, of from 0.1 mg to 140 mg per kilogram of body weight per day can be useful in the treatment of the above-indicated conditions (0.5 mg to 7 g per patient per day).
• the amount of active ingredient that may be combined with the vehicle to produce a single dosage form will vary depending upon the host treated and the particular mode of administration. Dosage unit forms will generally contain from 1 mg to 500 mg of an active ingredient.
• Frequency of dosage may also vary depending on the compound used and the particular disease treated. In some embodiments, for example, for the treatment of an allergic disorder and/or autoimmune and/or inflammatory disease, a dosage regimen of 4 times daily or less is used. In some embodiments, a dosage regimen of 1 or 2 times daily is used. It will be understood, however, that the specific dose level for any particular patient will depend upon a variety of factors including the activity of the specific compound employed, the age, body weight, general health, sex, diet, time of administration, route of administration, and rate of excretion, drug combination and the severity of the particular disease in the patient undergoing therapy.
• a labeled form of a chemical entity described herein can be used as a diagnostic for identifying and/or obtaining compounds that have the function of modulating an activity of a kinase as described herein.
• the chemical entities described herein may additionally be used for CTQ validating, optimizing, and standardizing bioassays.
• label herein is meant that the compound is either directly or indirectly labeled with a label which provides a detectable signal, e.g., radioisotope, fluorescent tag, enzyme, antibodies, particles such as magnetic particles, chemiluminescent tag, or specific binding molecules, etc.
• Specific binding molecules include pairs, such as biotin and streptavidin, digoxin and antidigoxin etc.
• the complementary member would normally be labeled with a molecule which provides for detection, in accordance with known procedures, as outlined above.
• the label can directly or indirectly provide a detectable signal.
• DMEM Dulbecco's modified Eagle's medium
• the resulting mixture was treated with tetrakis(triphenylphosphine)palladium(0) (1.20 g, 1.04 mmol) and stirred at reflux for 2.5 h. After this time, the reaction was cooled to ambient temperature, diluted with ethyl acetate (100 mL) and washed with brine (100 mL). The aqueous layer was extracted with ethyl acetate (100 mL) and the combined organic layers were dried over sodium sulfate, filtered and the filtrate was concentrated under reduced pressure.
• Electrospray MS is performed on a MICROMASS LCT equipped with a LockSpray source for accurate mass measurements. Spectra are acquired in positive ion mode from 100-1000 Da at an acquisition rate of 1 spectrum/0.9s with a 0.1s interscan delay. The instrument is tuned for a resolution of 5000 (FWHM). Every 5 th scan is taken from the reference position of the Lockspray source. Leucine enkephalin (556.2771 [M+H] + ) is used as the reference, or lock mass.
• a master mix minus Syk enzyme is prepared containing IX Cell Signaling kinase buffer (25 mM Tris-HCl, pH 7.5, 5 mM beta-glycerophosphate, 2 mM dithiothreitol, 0.1 raM Na 3 VO 4 , 10 mM MgCl 2 ), 0.5 ⁇ M Promega PTK Biotinylated peptide substrate 1, 0.01% casein, 0.01% Triton-XIOO, and 0.25% glycerol.
• IX Cell Signaling kinase buffer 25 mM Tris-HCl, pH 7.5, 5 mM beta-glycerophosphate, 2 mM dithiothreitol, 0.1 raM Na 3 VO 4 , 10 mM MgCl 2
• Promega PTK Biotinylated peptide substrate 1, 0.01% casein, 0.01% Triton-XIOO, and 0.25% glyce
• a master mix plus Syk enzyme is prepared containing IX Cell Signaling kinase buffer, 0.5 ⁇ M PTK Biotinylated peptide substrate 1, 0.01% casein, 0.01% Triton-XIOO, 0.25% glycerol and 0.4 ng/well Syk enzyme.
• Syk enzyme is purchased from Cell Signaling Technologies, expressed in baculovirus and is an N-terminally GST-tagged full length human wildtype Syk.
• the Syk nucleotide sequence accession number is NMJD03177, and accession number for the encoded Syk enzyme is NP_003168.
• the Syk protein was purified in one step using glutathione-agarose. The purity of the final protein preparation was assessed by SDS-PAGE and Coomassie staining.
• a solution of 200 ⁇ M ATP is prepared in water and adjusted to pH7.4 with IN NaOH.
• 5% DMSO is transferred to a 96- well Vi area Costar polystyrene plate.
• Compounds are tested singly and with an 11-point dose-responsive curve (starting concentration is 10 - 1 ⁇ M; 1:2 dilution).
• start concentration is 10 - 1 ⁇ M; 1:2 dilution.
• a quantity of 18.75 ⁇ L of master mix minus enzyme (as a negative control) and master mix plus enzyme is transferred to appropriate wells in 96-well Vi area costar polystyrene plate.
• 5 ⁇ L of 200 ⁇ M ATP is added to that mixture in the 96-well Vi area Costar polystyrene plate for final ATP concentration of 40 ⁇ M. The reaction is allowed to incubate for 1 hour at room temperature.
• the reaction is stopped with Perkin Elmer IX detection buffer containing 30 mM EDTA, 80 nM SA-APC, and 4 nM PT66 Ab.
• the plate is read using time-resolved fluorescence with a Perkin Elmer Envision using excitation filter 330 nm, emission filter 665 nm, and 2 nd emission filter 615 nm.
• IC 50 values are subsequently calculated using a log-logit regression algorithm.
• Ramos cells are serum starved at 2 x 10 6 cells/ml in serum-free RPMI for 1 hour in an upright T175 Falcon TC flask. Cells are centrifuged (1100 rpm x 5 min) and incubated at a density of 0.5xl0 7 cells/ml in the presence of test compound or DMSO controls for 1 hr at 37 °C.
• Cells are pelleted, lysed in 40 ul cell lysis buffer, and mixed with Invitrogen SDS-PAGE loading buffer. 20 ul of cell lysate for each sample are subject to SDS-PAGE and western blotting with anti-phosphoBLNK(Tyr96) antibody (Cell Signaling Technology #3601) to assess
• Syk activity and anti-Syk antibody (BD Transduction Labs #611116) to control for total protein load in each lysate.
• the images are detected using fluorescent secondary detection systems and the LiCor Odyssey instrument.
• a generalized procedure for a standard cellular B-cell proliferation assay that can be used to test compounds disclosed in this application is as follows.
• B-cells are purified from spleens of 8-16 week old Balb/c mice using a B-cell isolation kit (Miltenyi Biotech, Cat # 130-090-862). Test compounds are diluted in 0.25% DMSO and incubated with 2.5 x 10 5 purified mouse splenic B-cells for 30 min prior to addition of lO ⁇ g/ml of an anti-mouse IgM antibody (Southern Biotechnology Associates Cat # 1022-01) in a final volume of 100 ⁇ l.
• a generalized procedure for a standard T cell proliferation assay that can be used to test compounds disclosed in this application is as follows.
• T cells are purified from spleens of 8-16 week old Balb/c mice using a Pan T cell isolation kit (Miltenyi Biotech, Cat # 130-090-861). Test compounds are diluted in 0.25% DMSO and incubated with 2.5 x 10 5 purified mouse splenic T cells in a final volume of 100 ⁇ l in flat clear bottom plates precoated for 90 min at 37°C with 10 ⁇ g/ml each of anti-CD3 (BD # 553057) and anti-CD28 (BD # 553294) antibodies.
• a generalized procedure for a standard assay for the inhibition of B cell activity that can be used to test compounds disclosed in this application is as follows. [00156] Total mouse splenocytes are purified from spleens of 8-16 week old Balb/c mice by red blood cell lysis (BD Pharmingen #555899). Testing compounds are diluted to 0.5% DMSO and incubated with 1.25 x 10 6 splenocytes in a final volume of 200 ⁇ l in flat clear bottom plates (Falcon 353072) for 60 min at 37 0 C.
• Cells are then stimulated with the addition of 15 ⁇ g/ml IgM (Jackson ImmunoResearch 115-006-020), and incubated for 16 hr at 37 0 C, 5% CO 2 . Following the 16 hr incubation, cells are transferred to conical bottom clear 96-well plates and pelleted by centrifugation at 1200 x g'x 5 min. Cells are preblocked by CD16/CD32 (BD
• CD69-PE (BD Pharmingen #553237), and 7AAD (BD Pharmingen #51-6898 IE). Cells are sorted on a BD FACSCalibur and gated on the CD1977AAD " population. The levels of CD69 surface expression on the gated population is thus determined. If CD69 expression is determined following treatment with different concentrations of test compound, CD69 expression level can be plotted vbersus test compound concentration.
• BMMC bone-marrow derived mouse mast cell
• Bone-marrow derived mast cells were cultured for >4 weeks with IL-3 (10ng/ml) and SCF (lOng/ml). The cells were determined to be > 90% cKit + /FceRI + by FACS analysis at the time of use. Cells (6 x 10 7 cells/50 ml) were serum-starved in a T 150 tissue culture flask for
• test compounds (10OuI) are plated in a 96 well microtiter plate (Falcon 353072) and test compounds are serially diluted to a final concentration 0.25% DMSO in the plate for 1 hr at 37 0 C, 5% CO 2 .
• Wells are treated with a DNP-BSA antigen challenge (50 ng/ml) and incubated for and additional 30 min at
• Dose-response curves are generated using 4- parameter logistical fit and IC50s calculated.
• PCA Passive Cutaneous Anaphylaxis
• -DNP-BSA will be made up at 4mg/ml in sterile ddH 2 O and stored at 4O 0 C solution.
• DNP-BSA in 200 ul of 1% Evan's blue dye solution in saline. Tails are immersed in warm water prior to iv injection to improve success. 3) 30 minutes to 2 hr prior to this antigen challenge, drug is delivered sc or po in 10% EtOH/ 20% cremaphor/ 70% saline. 4) Animals are sacrifice by CO 2 inhalation 30-60 min post antigen challenge and ears are removed for extraction of Evan's blue dye in 500 ul of formamide overnight at 65 0 C. 5) Blood is obtained by cardiac puncture just prior to final cervical dislocation and processed for plasma to provide PK analysis. 6) Evan's blue dye is quantified by reading absorbency of 200 ul of extracted solution in microtiter plates at 620 nm. Study Design of Experiment
• Each animal has one IgE anti-DNP sensitized ear (right ear by convention) and one PBS control ear (left ear by convention).
• Groups 1-8 represent the vehicle and compound testing arms;
• Group 9 represents the non-antigen negative control;
• Group 10 represents the non-sensitized challenged negative control;
• Group 11 represents the non-antigen challenged, non-sensitized negative control group (Groups 9-11 represent negative controls for background levels only and require only minimal number of animals per group.)
• the compounds disclosed in the examples above were tested in the Syk biochemical assay described herein (Example 4) and certain of those compounds exhibited an IC 50 value less than or equal to 1 micromolar. Certain of those compounds exhibited an IC 5 0 value less than or equal to 100 nM. Certain of those compounds exhibited an IC 50 value less than or equal to 10 nM.
• Example 3 Some of the compounds disclosed in synthetic Example 3 were tested in the B- cell proliferation assay (as described in Example 6) and exhibited an IC 50 value less than or equal to 10 micromolar. Certain of those compounds exhibited an IC 5 0 value less than or equal to 1 micromolar.
• Certain compounds described herein exhibited IC 50 values for inhibition of T-cell proliferation that were at least 3 -fold, and in some instances 5-fold, greater than the IC 50 values of those compounds for inhibition of B-cell proliferation.
• B cell activity (under the conditions described in example 8), and exhibited an IC 5O value less than or equal to 10 micromolar. Certain of those compounds exhibited an IC 5O value less than or equal to 1 micromolar.
• Some of the compounds disclosed in described herein exhibited both biochemical and cell-based activity. For example, some of the compounds described herein exhibited an IC 5O value less than or equal to 10 micromolar in the Syk biochemical assay described herein (Example 4) and an IC 50 value less than or equal to 10 micromolar in at least one of the cell- based assays (other than the T-cell assay) described herein (Examples 5, 6, 8 or 9).
• Certain of those compounds exhibited an IC 5O value less than or equal to 1 micromolar in the Syk biochemical assay described herein (Example 4) and an IC 50 value less than or equal to 10 micromolar in at least one of the cell-based assays (other than the T-cell assay) described herein (Examples 5, 6, 8 or 9). Certain of those compounds exhibited an IC 50 value less than or equal to 0.1 micromolar and an IC 50 value less than or equal to 10 micromolar in at least one of the cell-based assays (other than the T-cell assay) described herein (Examples 5, 6, 8 or 9).

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