MODULATORS OF PROTEIN KINASES

Provided herein are small molecule protein kinase modulators, pharmaceutical compositions comprising such, and their uses in treating one or more conditions.

BACKGROUND

The more than 523 typical and atypical kinases in the human kinome represent a constellation of enzymes that catalyze the transfer of a phosphate group from ATP to a variety of amino acid residues, such as tyrosine, serine, and threonine. By so doing, these enzymes and their interrelated networks are effectors of cellular signal transduction. In particular, receptor tyrosine kinases (RTKs) coupled with their downstream intracellular kinases and phosphatases mediated cascades and feedback loops establish critical conduits for the transfer and regulation of signals from the cell exterior into the nucleus where transcriptional regulation takes place. Phosphate transfer to specific sites on proteins results in enzyme activation or inactivation, changes in conformation, increased or decreased affinity for other proteins, appropriate localization, and in some cases targeting of proteins for degradation by the proteosome. Kinase inhibitors, design strategies, and various mechanisms of inhibition have been extensively reviewed [Zhang J., et. al.Nature Reviews Cancer(2009) 9: 28-39; Blanc J. et. al., Anti-Cancer Agents in Med. Chem. (2013) 13, 17 pages; Gross S. et. al.,J. Clin. Invest.(2015) 125(5); 1780-9; Cosgarea I. et. al.,J. der Deutsch. Dermatol. Gesellschaft,(2017) 887-93, DOI: 10.1111/ddg.13321]. In addition, mechanistically similar lipid kinases, such as PI3Ks and SPK1, also contribute to the regulatory process (Brown J. R., et. al.,BMC Evolutionary Biology(2011) 11(4): 1471-2148; Alvarez S. E., et. al.,Nature(2010) 465: 1084-1088).

Because these processes regulate essential functions in cell growth, proliferation, differentiation and development, division, adhesion, angiogenesis, stress responses, cell-cell or cell-matrix interactions, short range contact-mediated axional guidance and mitogenesis, the activities of RTKs and their downstream kinase partners in signal transduction are tightly regulated and balanced through control of external receptor ligands as well as expression of receptors, receptor antagonists, decoy receptors, and through redundancies or crosstalk between signaling pathways. Therefore, the aberrant expression of kinases or activating mutations in kinases, inactivating mutations in negative regulators, and alterations in phosphatase expression or activity, are known to participate in a variety of diseases, including many cancers.

SUMMARY

Provided herein are compounds having the Formula I.

and pharmaceutically acceptable salts thereof, wherein R1, R2, R3, X, m, X1and q are as defined herein. These compounds act as modulators of protein kinase (e.g., kinase inhibitors) and are useful in treating conditions responsive to the inhibition of protein kinase (e.g., cancer). See e.g., Table 1 and 2.

In one aspect, the (4-fluorophenyl)pyrazolyl group on compounds of Formula I was found to be important for type II kinase inhibitors. It is contemplated that such compounds provide an alternative binding mode, compared to traditional type II inhibitors, which provides an alternative method for addressing kinase selectivity.

Also provided are pharmaceutically acceptable compositions comprising the disclosed protein kinase inhibitors.

DETAILED DESCRIPTION

1. General Description of Compounds

In a first embodiment, provided is a compound having the Formula I:

Alternatively, as part of a first embodiment, provided is a compound having the Formula I:

or a pharmaceutically acceptable salt thereof, wherein R3is —OR4, —NHR5, —N(C1-C6)alkylR5, —CCHR6, —NHCOR7, —N(C1-C6)alkylCOR7, —C(O)R7, phenyl, heteroaryl, or heterocyclyl, wherein said phenyl, heteroaryl and heterocyclyl are each optionally substituted with 1 to 3 groups selected from R8; or R3is taken together with one R2to form a 4- to 6-membered heteroaryl optionally substituted with a heteroaryl which is optionally substituted with 1 to 3 groups selected from R9; and q is 0, 1, or 2, wherein the remaining variables are as described in the preceding paragraph for Formula I.

When used in connection to describe a chemical group that may have multiple points of attachment, a hyphen (-) designates the point of attachment of that group to the variable to which it is defined. For example, —NRbRcmeans that the point of attachment for this group occurs on the nitrogen atom.

The terms “halo” and “halogen” refer to an atom selected from fluorine (fluoro, —F), chlorine (chloro, —Cl), bromine (bromo, —Br), and iodine (iodo, —I).

The term “alkyl” when used alone or as part of a larger moiety, such as “haloalkyl”, and the like, means saturated straight-chain or branched monovalent hydrocarbon radical. Unless otherwise specified, an alkyl group typically has 1-4 carbon atoms, i.e., (C1-C4)alkyl.

“Alkoxy” means an alkyl radical attached through an oxygen linking atom, represented by —O-alkyl. For example, “(C1-C4)alkoxy” includes methoxy, ethoxy, propoxy, and butoxy.

The term “haloalkyl” includes mono, poly, and perhaloalkyl groups where the halogens are independently selected from fluorine, chlorine, bromine, and iodine (e.g., —CF3, —CHF2, etc.

“Haloalkoxy” is a haloalkyl group which is attached to another moiety via an oxygen atom such as, e.g., but are not limited to —OCHF2or —OCF3.

The term “heteroaryl” used alone or as part of a larger moiety refers to a 5- to 12-membered (e.g., a 5- to 7-membered or 5- to 6-membered) aromatic radical containing 1-4 heteroatoms selected from N, O, and S. A heteroaryl group may be mono- or bi-cyclic. Monocyclic heteroaryl includes, for example, thienyl, furanyl, pyrrolyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, triazinyl, tetrazinyl, oxadiazolyl, thiazolyl, isothiazolyl, thiadiazolyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, etc. Bi-cyclic heteroaryls include groups in which a monocyclic heteroaryl ring is fused to one or more aryl or heteroaryl rings. Nonlimiting examples include indolyl, imidazopyridinyl, benzooxazolyl, benzoxadiazolyl, indazolyl, benzimidazolyl, benzthiazolyl, quinolyl, quinazolinyl, quinoxalinyl, pyrrolopyridinyl, pyrrolopyrimidinyl, pyrazolopyridinyl, thienopyridinyl, thienopyrimidinyl, indolizinyl, purinyl, naphthyridinyl, and pteridinyl. It will be understood that when specified, optional substituents on a heteroaryl group may be present on any substitutable position and, include, e.g., the position at which the heteroaryl is attached.

The term “heterocyclyl” means a 4- to 12-membered (e.g., a 4- to 7-membered or 4- to 6-membered) saturated or partially unsaturated heterocyclic ring containing 1 to 4 heteroatoms independently selected from N, O, and S. It can be monocyclic, bicyclic (e.g., a bridged, fused, or spiro bicyclic ring), or tricyclic. A heterocyclyl ring can be attached to its pendant group at any heteroatom or carbon atom that results in a stable structure. Examples of such saturated or partially unsaturated heterocyclic radicals include, without limitation, tetrahydrofuranyl, tetrahydrothienyl, tetrahydropyranyl, pyrrolidinyl, pyridinonyl, pyrrolidonyl, piperidinyl, oxazolidinyl, piperazinyl, dioxanyl, dioxolanyl, morpholinyl, dihydrofuranyl, dihydropyranyl, dihydropyridinyl, tetrahydropyridinyl, dihydropyrimidinyl, oxetanyl, azetidinyl and tetrahydropyrimidinyl. A heterocyclyl group may be mono- or bicyclic. The term “heterocyclyl” also includes, e.g., unsaturated heterocyclic radicals fused to another unsaturated heterocyclic radical or aryl or heteroaryl ring, such as for example, tetrahydronaphthyridine, indolinone, dihydropyrrolotriazole, imidazopyrimidine, quinolinone, dioxaspirodecane. It will also be understood that when specified, optional substituents on a heterocyclyl group may be present on any substitutable position and, include, e.g., the position at which the heterocyclyl is attached.

The term “spiro” refers to two rings that shares one ring atom (e.g., carbon).

The term “fused” refers to two rings that share two adjacent ring atoms with one another.

The term “bridged” refers to two rings that share three ring atoms with one another.

It is to be understood that, when a disclosed compound has at least one chiral center, the present invention encompasses one enantiomer free from the corresponding optical isomer, racemic mixture of the compound and mixtures enriched in one enantiomer relative to its corresponding optical isomer. When a mixture is enriched in one enantiomer relative to its optical isomers, the mixture contains, for example, an enantiomeric excess of at least 50%, 75%, 90%, 95% 99% or 99.5%.

The enantiomers of the present invention may be resolved by methods known to those skilled in the art, for example by formation of diastereoisomeric salts which may be separated, for example, by crystallization; formation of diastereoisomeric derivatives or complexes which may be separated, for example, by crystallization, gas-liquid or liquid chromatography; selective reaction of one enantiomer with an enantiomer-specific reagent, for example enzymatic esterification; or gas-liquid or liquid chromatography in a chiral environment, for example on a chiral support for example silica with a bound chiral ligand or in the presence of a chiral solvent. Where the desired enantiomer is converted into another chemical entity by one of the separation procedures described above, a further step is required to liberate the desired enantiomeric form. Alternatively, specific enantiomers may be synthesized by asymmetric synthesis using optically active reagents, substrates, catalysts or solvents, or by converting one enantiomer into the other by asymmetric transformation.

When a disclosed compound has at least two chiral centers, the present invention encompasses a diastereomer free of other diastereomers, a pair of diastereomers free from other diastereomeric pairs, mixtures of diastereomers, mixtures of diastereomeric pairs, mixtures of diastereomers in which one diastereomer is enriched relative to the other diastereomer(s) and mixtures of diastereomeric pairs in which one diastereomeric pair is enriched relative to the other diastereomeric pair(s). When a mixture is enriched in one diastereomer or diastereomeric pair(s) relative to the other diastereomers or diastereomeric pair(s), the mixture is enriched with the depicted or referenced diastereomer or diastereomeric pair(s) relative to other diastereomers or diastereomeric pair(s) for the compound, for example, by a molar excess of at least 50%, 75%, 90%, 95%, 99% or 99.5%.

The diastereoisomeric pairs may be separated by methods known to those skilled in the art, for example chromatography or crystallization and the individual enantiomers within each pair may be separated as described above. Specific procedures for chromatographically separating diastereomeric pairs of precursors used in the preparation of compounds disclosed herein are provided the examples herein.

The terms “subject” and “patient” may be used interchangeably, and means a mammal in need of treatment, e.g., companion animals (e.g., dogs, cats, and the like), farm animals (e.g., cows, pigs, horses, sheep, goats and the like) and laboratory animals (e.g., rats, mice, guinea pigs and the like). Typically, the subject is a human in need of treatment.

The term “inhibit,” “inhibition” or “inhibiting” includes a decrease in the baseline activity of a biological activity or process.

As used herein, the terms “treatment,” “treat,” and “treating” refer to reversing, alleviating, delaying the onset of, or inhibiting the progress of a disease or disorder, or one or more symptoms thereof, as described herein. In some aspects, treatment may be administered after one or more symptoms have developed, i.e., therapeutic treatment. In other aspects, treatment may be administered in the absence of symptoms. For example, treatment may be administered to a susceptible individual prior to the onset of symptoms (e.g., in light of a history of symptoms and/or in light of exposure to a particular organism, or other susceptibility factors), i.e., prophylactic treatment. Treatment may also be continued after symptoms have resolved, for example to delay their recurrence.

For use in medicines, the salts of the compounds described herein refer to non-toxic “pharmaceutically acceptable salts.” Pharmaceutically acceptable salt forms include pharmaceutically acceptable acidic/anionic or basic/cationic salts. Suitable pharmaceutically acceptable acid addition salts of the compounds described herein include e.g. salts of inorganic acids (such as hydrochloric acid, hydrobromic, phosphoric, nitric, and sulfuric acids) and of organic acids (such as, acetic acid, benzenesulfonic, benzoic, methanesulfonic, and p-toluenesulfonic acids). Compounds of the present teachings with acidic groups such as carboxylic acids can form pharmaceutically acceptable salts with pharmaceutically acceptable base(s). Suitable pharmaceutically acceptable basic salts include e.g., ammonium salts, alkali metal salts (such as sodium and potassium salts) and alkaline earth metal salts (such as magnesium and calcium salts). Compounds with a quaternary ammonium group also contain a counteranion such as chloride, bromide, iodide, acetate, perchlorate and the like. Other examples of such salts include hydrochlorides, hydrobromides, sulfates, methanesulfonates, nitrates, benzoates and salts with amino acids such as glutamic acid.

The term “effective amount” or “therapeutically effective amount” refers to an amount of a compound described herein that will elicit a desired or beneficial biological or medical response of a subject e.g., a dosage of between 0.01-100 mg/kg body weight/day.

3. Description of Exemplary Compounds

In a second embodiment, the compound of Formula I is of the Formula II:

or a pharmaceutically acceptable salt thereof, wherein the variables are as described above for Formula I.

In a third embodiment, the compound of Formula I is of the Formula III:

or a pharmaceutically acceptable salt thereof, wherein the variables are as described above for Formula I.

In a fourth embodiment, the compound of Formula I is of the Formula IV:

or a pharmaceutically acceptable salt thereof, wherein the variables are as described above for Formula I.

In a fifth embodiment, R2in the compound of any one of Formulae I to IV, or a pharmaceutically acceptable salt thereof, is —SO2Ra, —SORa, or —SRa, wherein the remaining variables are as described above for Formula I. Alternatively, as part of a fifth embodiment, R2in the compound of any one of Formulae I to IV, or a pharmaceutically acceptable salt thereof, is —SO2Ra, —SORa, or —SRaand Rais (C1-C3)alkyl, wherein the remaining variables are as described above for Formula I. In another alternative, as part of a fifth embodiment, R2in the compound of any one of Formulae I to IV, or a pharmaceutically acceptable salt thereof, is —SO2CH3, —SCH3, or —SOCH3.

In a sixth embodiment, R2in the compound of any one of Formulae I to IV, or a pharmaceutically acceptable salt thereof, is halogen, hydroxyl, —NRbRd, or (C1-C3)alkyl, wherein the remaining variables are as described above for Formula I or the fifth embodiment. Alternatively, as part of a sixth embodiment, R2in the compound of any one of Formulae I to IV, or a pharmaceutically acceptable salt thereof, is hydroxyl, fluoro, bromo, methyl or NH2, wherein the remaining variables are as described above for Formula I or the fifth embodiment.

In a seventh embodiment, q in the compound of any one of Formulae I to IV, or a pharmaceutically acceptable salt thereof, is 1 or 2, wherein the remaining variables are as described above for Formula I or the fifth or sixth embodiment. In a seventh embodiment, q in the compound of any one of Formulae I to IV, or a pharmaceutically acceptable salt thereof, is 1 or 2, wherein the remaining variables are as described above for Formula I or the fifth or sixth embodiment.

In an eighth embodiment, R3in the compound of any one of Formulae I to IV, or a pharmaceutically acceptable salt thereof, is —OR4, NHR5, —CCHR6, —NHCOR7, —C(O)R7, phenyl, heteroaryl, or heterocyclyl, wherein said phenyl, heteroaryl and heterocyclyl are each optionally substituted with 1 to 3 groups selected from R8; or R3is taken together with one R2to form a 4- to 6-membered heteroaryl optionally substituted with a 9- or 10-membered fused-bicyclic heteroaryl which is optionally substituted with 1 to 3 groups selected from R9, wherein the remaining variables are as described above for Formula I or the fifth, sixth, or seventh embodiment. Alternatively, as part of an eighth embodiment, R3in the compound of any one of Formulae I to IV, or a pharmaceutically acceptable salt thereof, is —OR4, NHR5, —CCHR6, —NHCOR7, —C(O)R7, phenyl, 5- to 6-membered heteroaryl, 9- to 10-membered fused bicyclic heteroaryl, 5- to 6-membered heterocyclyl, or 9- to 10-membered fused bicyclic heterocyclyl, wherein said phenyl, 5- to 6-membered heteroaryl, 9- to 10-membered fused bicyclic heteroaryl, 5- to 6-membered heterocyclyl, and 9- to 10-membered fused bicyclic heterocyclyl are each optionally substituted with 1 to 3 groups selected from R8; or R3is taken together with one R2to form a 5-membered heteroaryl optionally substituted with a 9- or 10-membered fused-bicyclic heteroaryl which is optionally substituted with 1 to 3 groups selected from R9, wherein the remaining variables are as described above for Formula I or the fifth, sixth, or seventh embodiment. In another alternative, as part of an eighth embodiment, R3in the compound of any one of Formulae I to IV, or a pharmaceutically acceptable salt thereof, is —OR4, NHR5, —CCHR6, —NHCOR7, —C(O)R7, (C1-C4)alkylpyrrolopyridinyl, phenyl, pyridinyl, thienopyrimidinyl, dihydropyridopyrimidinyl, dihydrobenzoimidazolyl imidazopyridinyl, pyridopyrimidinyl, or dihydropyridinyl, wherein said phenyl, pyridinyl, thienopyrimidinyl, dihydropyridopyrimidinyl, dihydrobenzoimidazolyl imidazopyridinyl, pyridopyrimidinyl, dihydropyridinyl, and pyrrolopyridinyl on the (C1-C4)alkylpyrrolopyridinyl are each optionally substituted with 1 to 3 groups selected from R8; or R3is taken together with one R2to form a furanyl optionally substituted with imidazopyridazinyl which is optionally substituted with 1 to 3 groups selected from R9, wherein the remaining variables are as described above for Formula I or the fifth, sixth, or seventh embodiment. In another alternative, as part of an eighth embodiment, R3in the compound of any one of Formulae I to IV, or a pharmaceutically acceptable salt thereof, is —OR4, NHR5, —CCHR6, —NHCOR7, —C(O)R7, phenyl, pyridinyl, thienopyrimidinyl, dihydrobenzoimidazolyl imidazopyridinyl, pyridopyrimidinyl, or dihydropyridinyl, wherein said phenyl, pyridinyl, thienopyrimidinyl, dihydrobenzoimidazolyl imidazopyridinyl, pyridopyrimidinyl, and dihydropyridinyl are each optionally substituted with 1 to 3 groups selected from R8; or R3is taken together with one R2to form a furanyl optionally substituted with imidazopyridazinyl which is optionally substituted with 1 to 3 groups selected from R9, wherein the remaining variables are as described above for Formula I or the fifth, sixth, or seventh embodiment.

In a ninth embodiment, R4in the compound of any one of Formulae I to IV, or a pharmaceutically acceptable salt thereof, is 5- to 6-membered heteroaryl or 9- to 10-membered fused bicyclic heteroaryl, each of which are optionally substituted with 1 to 3 groups selected from R10, wherein the remaining variables are as described above for Formula I or the fifth, sixth, seventh, or eighth embodiment. Alternatively, as part of a ninth embodiment, R4in the compound of any one of Formulae I to IV, or a pharmaceutically acceptable salt thereof, is pyridinyl, dihydroquinazolinyl, dihydropyridopyrimidinyl, thiazolopyridinyl, quinolinyl, pyrrolopyridinyl, or tetrahydronaphthyridinyl, each of which are optionally substituted with 1 to 3 groups selected from R10, wherein the remaining variables are as described above for Formula I or the fifth, sixth, seventh, or eighth embodiment. In another alternative, as part of a ninth embodiment, R4in the compound of any one of Formulae I to IV, or a pharmaceutically acceptable salt thereof, is pyridinyl, dihydroquinazolinyl, dihydropyridopyrimidinyl, thiazolopyridinyl, quinolinyl, or tetrahydronaphthyridinyl, each of which are optionally substituted with 1 to 3 groups selected from R10, wherein the remaining variables are as described above for Formula I or the fifth, sixth, seventh, or eighth embodiment.

In a tenth embodiment, R5in the compound of any one of Formulae I to IV, or a pharmaceutically acceptable salt thereof, is 5- to 6-membered heteroaryl, 9- to 10-membered fused bicyclic heteroaryl, 5- to 6-membered heterocyclyl, or 9- to 10-membered fused bicyclic heterocyclyl, each of which are optionally substituted with 1 to 3 groups selected from R11, wherein the remaining variables are as described above for Formula I or any one of the fifth to ninth embodiments. Alternatively, as part of a tenth embodiment, R5in the compound of any one of Formulae I to IV, or a pharmaceutically acceptable salt thereof, is pyridinyl, pyrimidinyl, dihydroquinazolinyl, thiazolyl, dihydropyridopyrimidinyl, or imidazopyridazinyl, each of which are optionally substituted with 1 to 3 groups selected from R11, wherein the remaining variables are as described above for Formula I or any one of the fifth to ninth embodiments.

In an eleventh embodiment, R6in the compound of any one of Formulae I to IV, or a pharmaceutically acceptable salt thereof, is a 9- to 10-membered fused bicyclic heteroaryl optionally substituted with 1 to 3 groups selected from R12, wherein the remaining variables are as described above for Formula I or any one of the fifth to tenth embodiments. Alternatively, as part of an eleventh embodiment, R6is imidazopyridazinyl or thienopyrimidinyl, each of which are optionally substituted with 1 to 3 groups selected from R12, wherein the remaining variables are as described above for Formula I or any one of the fifth to tenth embodiments.

In a twelfth embodiment, R7in the compound of any one of Formulae I to IV, or a pharmaceutically acceptable salt thereof, is a 5- to 6-membered heteroaryl or a 9- to 10-membered heteroaryl each optionally substituted with 1 to 3 groups selected from R12, wherein the remaining variables are as described above for Formula I or any one of the fifth to eleventh embodiments. Alternatively, as part of a twelfth embodiment, R7in the compound of any one of Formulae I to IV, or a pharmaceutically acceptable salt thereof, is a 5- to 6-membered heteroaryl optionally substituted with 1 to 3 groups selected from R12, wherein the remaining variables are as described above for Formula I or any one of the fifth to eleventh embodiments. In another alternative, as part of a twelfth embodiment, R7in the compound of any one of Formulae I to IV, or a pharmaceutically acceptable salt thereof, is pyrrolopyridinyl, wherein the remaining variables are as described above for Formula I or any one of the fifth to eleventh embodiments. In another alternative, as part of a twelfth embodiment, R7in the compound of any one of Formulae I to IV, or a pharmaceutically acceptable salt thereof, is a 9- to 10-membered heteroaryl optionally substituted with 1 to 3 groups selected from R12.

In a thirteenth embodiment, R8in the compound of any one of Formulae I to IV, or a pharmaceutically acceptable salt thereof, is selected from (C1-C6)alkyl, oxo, morpholinyl, —O(C1-C6)hydroxyalkyl, and —NRbC(O)Rb, NRbRc, wherein the remaining variables are as described above for Formula I or any one of the fifth to twelfth embodiments.

In a fourteenth embodiment, R9in the compound of any one of Formulae I to IV, or a pharmaceutically acceptable salt thereof, is halo, wherein the remaining variables are as described above for Formula I or any one of the fifth to thirteenth embodiments.

In a fifteenth embodiment, R10in the compound of any one of Formulae I to IV, or a pharmaceutically acceptable salt thereof, is —C(O)NRbRc, —NRbRc, (C1-C6)alkoxy, and pyridinyl, wherein said pyridinyl is optionally substituted with —(C1-C6)alkylNH(C1-C6)hydroxyalkyl, wherein the remaining variables are as described above for Formula I or any one of the fifth to fourteenth embodiments.

In a sixteenth embodiment, R11in the compound of any one of Formulae I to IV, or a pharmaceutically acceptable salt thereof, is oxo, (C1-C6)alkyl, and heteroaryl, wherein said heteroaryl is optionally substituted with 1 to 3 groups selected from NRbRc, halo, (C1-C6)alkyl, —NRbC(O)Rb, and —NRbC(O)ORb, wherein the remaining variables are as described above for Formula I or any one of the fifth to fifteenth embodiments. Alternatively, as part of a sixteenth embodiment, R11in the compound of any one of Formulae I to IV, or a pharmaceutically acceptable salt thereof, is oxo, (C1-C6)alkyl, pyridinyl, thiazolyl, or purinyl, wherein said pyridinyl, thiazolyl, or purinyl are each optionally substituted with 1 to 3 groups selected from NRbRc, halo, (C1-C6)alkyl, —NRbC(O)Rb, and —NRbC(O)ORb, wherein the remaining variables are as described above for Formula I or any one of the fifth to fifteenth embodiments. In another alternative, as part of a fifteenth embodiment, R11in the compound of any one of Formulae I to IV, or a pharmaceutically acceptable salt thereof, is oxo, (C1-C6)alkyl, pyridinyl, thiazolyl, or purinyl, wherein said pyridinyl, thiazolyl, or purinyl are each optionally substituted with 1 to 3 groups selected is optionally substituted with 1 to 3 groups selected from oxo and (C1-C6)alkyl, wherein the remaining variables are as described above for Formula I or any one of the fifth to fifteenth embodiments.

In a seventeenth embodiment, R12in the compound of any one of Formulae I to IV, or a pharmaceutically acceptable salt thereof, is NRbRcor halo, wherein the remaining variables are as described above for Formula I or any one of the fifth to fifteenth embodiments.

In an eighteenth embodiment, Rcin the compound of any one of Formulae I to IV, or a pharmaceutically acceptable salt thereof, is selected from hydrogen, (C1-C6)alkyl, and 4- to 6-membered heterocyclyl, wherein the remaining variables are as described above for Formula I or any one of the fifth to seventeenth embodiments.

Compounds having the disclosed formulae are further disclosed in the Exemplification and are included in the present disclosure. Pharmaceutically acceptable salts thereof as well as the neutral forms are included.

4. Uses, Formulation and Administration

The compounds and compositions described herein are generally useful for modulating the activity of protein kinase. In some aspects, the compounds and pharmaceutical compositions described herein inhibit the activity of protein kinase.

In some aspects, the compounds and pharmaceutical compositions described herein are useful in treating a disorder associated with protein kinase function. Thus, provided herein are methods of treating a condition associated with protein kinase function, comprising administering to a subject in need thereof, a therapeutically effective amount of a compound described herein, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising a disclosed compound or pharmaceutically acceptable salt thereof. Also provided is the use of a compound described herein, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising a disclosed compound or pharmaceutically acceptable salt thereof, for the manufacture of a medicament for treating a condition associated with protein kinase function. Also provided is a compound described herein, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising a disclosed compound or pharmaceutically acceptable salt thereof, for use in treating a condition associated with protein kinase function.

In some aspects, the compounds and pharmaceutical compositions described herein are useful in treating a condition selected from an inflammatory disease, a neurodegenerative disease, cardiovascular disease, metabolic disease, pain, and cancer.

Examples of neurodegenerative disease include, but are not limited to, Alzheimers, Parkinson's disease, and multiple sclerosis.

Examples of cardiovascular disease include, but are not limited to, hypertension, coronary and cerebral vasospasm, restenosis, atherosclerosis, stroke, and heart failure

Examples of metabolic disease include, but are not limited to, type 1 diabetes, type 2 diabetes.

In certain aspects, a pharmaceutical composition described herein is formulated for administration to a patient in need of such composition. Pharmaceutical compositions described herein may be administered orally, parenterally, by inhalation spray, topically, rectally, nasally, buccally, vaginally or via an implanted reservoir. The term “parenteral” as used herein includes subcutaneous, intravenous, intramuscular, intra-articular, intra-synovial, intrasternal, intrathecal, intrahepatic, intralesional and intracranial injection or infusion techniques. In some embodiments, the compositions are administered orally, intraperitoneally or intravenously. Sterile injectable forms of the pharmaceutical compositions described herein may be aqueous or oleaginous suspension. These suspensions may be formulated according to techniques known in the art using suitable dispersing or wetting agents and suspending agents.

In some aspects, the pharmaceutical compositions are administered orally.

Kinase compounds disclosed herein are synthesized according to the following examples. As used below, and throughout the description of the invention, the following abbreviations, unless otherwise indicated, shall be understood to have the following meanings:ACN: acetonitrile° C.: degrees Celsiusδ: chemical shift in parts per million downfield from tetramethylsilanedichloromethane (CH2Cl2)DCM: dimethylformamideDMF: dimethylsulfoxideEt2O: diethyl etherEtOAc: ethyl acetateES+: electrospray ionizationEt: ethylg: gram(s)Hex: hexanesh: hour(s)HPLC: high performance liquid chromatographyHz: hertzJ: coupling constant (in NMR spectrometry)LCMS: liquid chromatography mass spectrometryp: microm: multiplet (spectral); meter(s); milliM: molarM+: parent molecular ionMe: methylMeOH: methanolMHz: megahertzmin: minute(s)mol: mole(s); molecular (as in mol wt)mL: milliliterNIS: N-iodosuccinimideMS: mass spectrometrynm: nanometer(s)NMR: nuclear magnetic resonancepH: negative base 10 logarithm of hydrogen cation concentration; a measure of the acidity or basicity of an aqueous solutionPE: petroleum etherrt: room temperatures: singlet (spectral)t: triplet (spectral)T: temperatureTFA: trifluoroacetic acidTHF: tetrahydrofuran.

General Analytical Techniques

Liquid Chromatography Mass Spectrometry (LCMS) was performed on a Shimadzu LCMS system consisting of Nexera XR HPLC stack (20 Series) with Nexera X2 SPD-M30A DAD and LCMS-2020 mass spectrometer using LabSolutions, v.5.89 software under the following parameters: Column temp: 45° C., Sample temp: 18° C. Gradient elution methods, mobile phase eluents, and columns are shown below.

Time (min)Solvent A (%)Solvent B (%)0.09500.295010.2010012.2010012.495514.0955

Time (min)Solvent A (%)Solvent B (%)0.010000.2100010.259512.559513.0100014.51000

Time (min)Solvent A (%)Solvent B (%)0.09550.295510.2010012.2010012.495514.5955

Time (min)Solvent A (%)Solvent B (%)0.09550.295510.2010012.2010012.495514.2955

Time (min)Solvent A (%)Solvent B (%)09550.19553.601004.601004.7955

Method B

Time (min)Solvent A (%)Solvent B (%)09550.019550.5001001.4995540100

Time (min)Solvent A (%)Solvent B (%)09550.019559.6010010.6010010.7955

Neutral Mobile Phase

Acidic Mobile Phase

Solvent A (0.1% formic acid in water, pH 2.3)Solvent B (0.1% formic acid in acetonitrile)

Columns

Method C

Liquid Chromatography Mass Spectrometry (LCMS) was performed on a Shimadzu LCMS system consisting of consisting of a LC 20 AD prominence pump, DGU-20 A3 prominence degasser, SPD-M20A prominence DAD detector, SIL-HTC autosampler coupled with a Shimadzu LCMS (SQD) mass spectrometer using Lab Solutions, v.3.70.390 software under the following parameters: Column temp: 50° C. Gradient elution methods, mobile phase eluents, PDA detection and columns are shown below.

Time (min)Solvent A (%)Solvent B (%)0.09822.02983.02983.209824.0982

Acidic Mobile Phase

Columns

Method D

Liquid Chromatography Mass Spectrometry (LCMS) was performed on a Shimadzu LCMS system consisting of a LC 20 AD prominence pump, DGU-20 A3 prominence degasser, SPD-M20A prominence DAD detector, SIL-HTC autosampler coupled with a Shimadzu LCMS (SQD) mass spectrometer using Lab Solutions, v.3.70.390 software under the following parameters: Column temp: 50° C. Gradient elution methods, mobile phase eluents, PDA detection and columns are shown below.

Time (min)Solvent A (%)Solvent B (%)0.010006.02988.02989.0100010.01000

Acidic Mobile Phase

Columns

Method E

Liquid Chromatography Mass Spectrometry (LCMS) was performed on a Shimadzu LCMS system consisting of a LC 20 AD prominence pump, DGU-20 A3 prominence degasser, SPD-M20A prominence DAD detector, SIL-HTC autosampler coupled with a Shimadzu LCMS(SQD) mass spectrometer using Lab Solutions, v.3.70.390 software under the following parameters: Column temp: 50° C. Gradient elution methods, mobile phase eluents, PDA detection and columns are shown below.

Time (min)Solvent A (%)Solvent B (%)0.019820.29822.22983.02983.29824.0982

Acidic Mobile Phase

Columns

Method F

Liquid Chromatography Mass Spectrometry (LCMS) was performed on a Shimadzu LCMS system consisting of consisting of a LC 20 AD prominence pump, DGU-20 A3 prominence degasser, SPD-M20A prominence DAD detector, SIL-HTC autosampler coupled with a Shimadzu LCMS(SQD) mass spectrometer using Lab Solutions, v.3.70.390 software under the following parameters: Column temp: 40° C. Gradient elution methods, mobile phase eluents, PDA detection and columns are shown below.

Time (min)Solvent A (%)Solvent B (%)0.0010001.501002.201002.610003.0937

Mobile Phase

Columns

X Select CSH C18 (3.0*50) mm 2.5 u

Analytical HPLC

Method A

HPLC analyses were obtained on a Shimadzu HPLC 2010CHT HPLC consisting of a LC 20 AD prominence pump, DGU-20 A3 prominence degasser, SPD-M20A prominence DAD detector, SIL-HTC autosampler using LC Solutions, v.1.25 software under the following parameters: Column temp: 35° C. Gradient elution methods, mobile phase eluents, PDA detection and columns are shown below.

Time (min)Solvent A (%)Solvent B (%)0.019551.09558.0010012.0010014.095518.0955

Mobile Phase

Columns

Method B

HPLC analyses were obtained on a Shimadzu HPLC 2010CHT HPLC consisting of a LC 20 AD prominence pump, DGU-20 A3 prominence degasser, SPD-M20A prominence DAD detector, SIL-HTC autosampler using LC Solutions, v.1.25 software under the following parameters: Column temp: 35° C. Gradient elution methods, mobile phase eluents, PDA detection and columns are shown below.

Time (min)Solvent A (%)Solvent B (%)0.0190106.0109010.0010012.0010014.0901018.09010

Mobile Phase

Columns

Method C

HPLC analyses were obtained on a Shimadzu HPLC 2010CHT HPLC consisting of a LC 20 AD prominence pump, DGU-20 A3 prominence degasser, SPD-M20A prominence DAD detector, SIL-HTC autosampler using LC Solutions, v.1.25 software under the following parameters: Column temp: 35° C. Gradient elution methods, mobile phase eluents, PDA detection and columns are shown below.

Time (min)Solvent A (%)Solvent B (%)0.0190106.0109010.0010012.0010014.0901018.09010

Mobile Phase

Columns

Method D

HPLC analyses were obtained on a Shimadzu HPLC 2010CHT HPLC consisting of a LC 20 AD prominence pump, DGU-20 A3 prominence degasser, SPD-M20A prominence DAD detector, SIL-HTC autosampler using LC Solutions, v.1.25 software under the following parameters: Column temp: 35° C. Gradient elution methods, mobile phase eluents, PDA detection and columns are shown below.

Time (min)Solvent A (%)Solvent B (%)0.019551.09558.0010012.0010014.095518.0955

Mobile Phase

Columns

Neutral Mobile Phase

Acidic Mobile Phase

Columns

Method A

HPLC analyses were obtained on a Shimadzu HPLC 2010CHT HPLC consisting of a LC 20 AD prominence pump, DGU-20 A3 prominence degasser, SPD-M20A prominence DAD detector, SIL-HTC autosampler using LC Solutions, v.1.25 software under the following parameters: Column temp: 35° C. Gradient elution methods, mobile phase eluents, PDA detection and columns are shown below.

Time (min)Solvent A (%)Solvent B (%)0.019551.09558.0010012.0010014.095518.0955

Mobile Phase

Columns

Method B

HPLC analyses were obtained on a Shimadzu HPLC 2010CHT HPLC consisting of a LC 20 AD prominence pump, DGU-20 A3 prominence degasser, SPD-M20A prominence DAD detector, SIL-HTC autosampler using LC Solutions, v.1.25 software under the following parameters: Column temp: 35° C. Gradient elution methods, mobile phase eluents, PDA detection and columns are shown below.

Solubility

Mobile Phase

Columns

Method C

HPLC analyses were obtained on a Shimadzu HPLC 2010CHT HPLC consisting of a LC 20 AD prominence pump, DGU-20 A3 prominence degasser, SPD-M20A prominence DAD detector, SIL-HTC autosampler using LC Solutions, v.1.25 software under the following parameters: Column temp: 35° C. Gradient elution methods, mobile phase eluents, PDA detection and columns are shown below.

Time (min)Solvent A (%)Solvent B (%)0.019551.09558.0010012.0010014.095518.0955

Mobile Phase

Method D

HPLC analyses were obtained on a Shimadzu HPLC 2010CHT HPLC consisting of a LC 20 AD prominence pump, DGU-20 A3 prominence degasser, SPD-M20A prominence DAD detector, SIL-HTC autosampler using LC Solutions, v.1.25 software under the following parameters: Column temp: 35° C. Gradient elution methods, mobile phase eluents, PDA detection and columns are shown below.

Time (min)Solvent A (%)Solvent B (%)0.019551.09558.0010012.0010014.095518.0955

Mobile Phase

X-Bridge C18 (4.6*150) mm 5 u1H NMR Proton NMR was performed on the Varian Inova 500 spectrometer operating at 500 MHz in CDCl3, DMSO-d6, or MeOD.or1H NMR Proton NMR was performed on the Varian MR 400 MHz or Bruker Avance neo 400 MHz spectrometer operating at 400 MHz in CDCl3, DMSO-d6, or MeOD.

Synthetic Method A: Typically used 1 equivalent of amine intermediate, 1-2 equivalents of acid intermediate, 1 to 2 equivalents of HATU, 2-4 equivalents of DIPEA, and 3 to 16 hr of reaction time in DMF. A full example is written out below for Example 4

Synthetic Method B: Typically used 1 equivalent of amine intermediate, A range of 1-2 equivalents of acid intermediate, A range 1 to 2 equivalents of HATU, A range of 2-4 equivalents of DIPEA, and reaction time range of 3 to 16 hr in DMF. Followed by a deprotection step which used with TFA or HCl for deprotection of a Boc or THP protection group. A full example is written out below for Example 8

Synthesis of [Example 1]: Followed the same procedure as Method A

Synthesis of Example 3 followed the same procedure used in Synthetic Method A except used 1-(4-fluorophenyl)-5-(methylthio)-1H-pyrazole-3-carboxylic acid and 6-((5-amino-2-methylphenyl)amino)-3-methylquinazolin-4(3H)-one. The solid was collected by filtration and dried to afford 55 mg, 0.107 mmol, 17.97% yield of the title compound as an off-white solid. MS(ES+) m/z calc'd for [M+H]+[C27H23FN6O2S+H]+: 515.2, found 515.2, tR=2.38 min [Analytical Method B].

A solution tert-butyl 3-(4-(3-amino-4-fluorophenoxy)picolinamido)azetidine-1-carboxylate (0.15 g, 0.37 mmol), 1-(4-fluorophenyl)-5-(methylthio)-1H-pyrazole-3-carboxylic acid (0.100 g, 0.41 mmol), and HATU (0.209 g, 0.55 mmol) in DMF (3.73 mL) was added DIEA (0.129 mL, 0.74 mmol) dropwise at rt. Then the mixture was stirred at rt for 16 h. At the end of the period water was added and the solid separated was collected and washed with sat. NaHCO3solution followed by water. The crude was chromatographed over silica gel using gradient of EtOAc in DCM to afford title product. Yield: 0.124 g

Synthesis of Example 16 followed the same procedure as Synthetic Method A except used 4-(4-amino-3-fluoro-phenoxy)-N-methyl-pyridine-2-carboxamide and 1-(4-fluorophenyl)-3-(methylsulfinyl)-1H-pyrazole-5-carboxylic acid. The crude was chromatographed over silica gel using 0-15% MeOH in DCM to afford the title compound.

Synthesis of Example 17 followed the same procedure as Synthetic Method A except used 4-(4-amino-3-fluoro-phenoxy)-N-methyl-pyridine-2-carboxamide and 1-(4-fluorophenyl)-5-(methylsulfonyl)-1H-pyrazole-3-carboxylic acid. The crude was chromatographed over silica gel using 0-15% MeOH in DCM to afford the title compound.

General Synthetic Method C:

Synthesis of N-(2-fluoro-5-((3-methyl-4-oxo-3,4-dihydroquinazolin-6-yl)oxy)phenyl)-1-(4-fluorophenyl)-1H-pyrazole-3-carboxamide [Example 80]: Method C

Synthesis of Example 7 Followed the Same Procedure Used in Synthetic Method B Except Used

6-(3-amino-2,6-dichloro-phenyl)-8-methyl-2-(methylamino)pyrido[2,3-d]pyrimidin-7-one and 1-(4-fluorophenyl)-1H-pyrazole-3-carboxylic acid. The crude compound was purified by preparative HPLC purification Method A. The preparative fractions were lyophilized to get solid compound which was dried under high vacuum to get the title compound (2.19 mg, 0.004 mmol, 4.71% yield) as an off-white solid. MS (ES+) calc'd for [M+H]+[C25H18Cl2FN7O2+H]+:539.36, found: 538.08, LCMS: tR: 2.10 min. [Analytical Method: D].

Synthesis of Example 13 Followed the Same Procedure Used in Synthetic Method E Except Used

Synthesis of Example 21 Followed the Same Procedure Used in Synthetic Method B Except Used

Synthesis of Example 24 Followed the Same Procedure Used in Synthetic Method B Except Used

Hinge Binding Intermediates

Synthesis of Hinge Intermediates:

Available from Commercial Sources

Available from Commercial Sources

Intermediate 3 is known in the literature and the synthesis can be performed according to the reference below:Reference: WO2006/24834A1

Intermediate 4 is known in the literature and the synthesis can be performed according to the reference below:Reference: KR2020/88945, 2020, A

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Intermediate 7 is known in the literature and the synthesis can be performed according to the reference below:Reference: Journal of Medicinal Chemistry, 2020, vol. 63, #5, p. 2013-2027

Intermediate 8 is known in the literature and the synthesis can be performed according to the reference below:Reference: Journal of Medicinal Chemistry, 2020, vol. 63, #5, p. 2013-2027

Intermediate 11 is known in the literature and the synthesis can be performed according to the reference below:Reference: CN-103102349-B

Intermediate 12 is known in the literature and the synthesis can be performed according to the reference below:Reference: Chemical Biology and Drug Design, 2019, vol. 94, #6, p. 2013-2022

Intermediate 14 is known in the literature and the synthesis can be performed according to the reference below:Reference: US2019/169166, 2019, A1

Intermediate is known in the literature and the synthesis can be performed according to the reference below:Reference: WO2006/24834A1

A flame dried pressure vessel containing ethynyl(trimethyl)silane (4.1 mL, 28.8 mmol, 3.50 eq), 3-iodoimidazo[1,2-b]pyridazine (2.01 g, 8.22 mmol, 1.00 eq), Pd(PPh3)2Cl2(0.29 g, 0.411 mmol, 0.0500 eq), and Et3N (2.5 mL, 18.1 mmol, 2.20 eq) in THF (0.2M, 41 mL) was purged with nitrogen at rt for 10 minutes prior to the addition of CuI (0.047 g, 0.247 mmol, 0.0300 eq). The resulting mixture was lowered into an oil bath at 50° C. and stirred overnight. The reaction was cooled to rt, additional ethynyl(trimethyl)silane (1.5 equiv., 1.76 mL) added, headspace purged with nitrogen, and lowered back into oil bath to stir overnight again at 50° C. The reaction mixture was concentrated in vacuo. The resulting residue was dissolved in MeOH (100 mL), K2CO3(0.2 equiv., 0.227 g) was added, and the resulting reaction mixture was stirred at rt overnight. Began heating the mixture to 50° C. the following morning. After 2.66 hours additional K2CO3(0.8 equiv., 0.909 g) was added and the reaction mixture was heated further to reflux. After 2.5 hours the reaction was cooled to rt, concentrated in vacuo, and the resulting residue taken up in EtOAc and water prior to filtering through celite (rinsing with EtOAc). Aqueous was extracted further with EtOAc and combined organics were rinsed with brine and dried over sodium sulfate before concentrating in vacuo to afford 0.99 g, 6.90 mmol crude title compound as a solid which was used directly in the subsequent step.1H NMR (600 MHz, DMSO-d6) δ 8.65 (dd, J=4.4, 1.5 Hz, 1H), 8.21 (dd, J=9.2, 1.5 Hz, 1H), 8.11 (s, 1H), 7.35 (dd, J=9.2, 4.4 Hz, 1H), 4.95 (s, 1H); MS (ES+) m/z calc'd for [M+H]+[C8H5N3+H]+: 144.1 found 144.2, LCMS: tR=3.00 min [Analytical Method: 05991008_AA1.lcm]

Intermediate 17 is known in the literature and the synthesis can be performed according to the reference below:Reference: WO2007/2325A1

Intermediate 18 is known in the literature and the synthesis can be performed according to the reference below:Reference: Journal of Medicinal Chemistry, 2020, vol. 63, #5, p. 2013-2027

A solution of DMFDMA (40 mL, 80.0 mmol, 1.00 eq) in 2-Amino-4-methyl-5-acetylthiazole (12.50 g, 80.0 mmol, 1.00 eq) was heated to 100° C. for 16 h. After cooling to rt the reaction mixture is concentrated in vacuo and the crude was triturated from ethyl acetate and filtered through a fritted Buchner funnel resulting in 10.5 g the title compound as a light orange solid. The mother liquor was concentrated in vacuo and the crude solid was recrystallized from EtOAc which resulted in an additional 0.675 g. The total yield was 11.2 g, 52.5% yield of the title compounds as a light orange solid.1H NMR (600 MHz, DMSO-d6) δ 8.35 (s, 1H), 7.56 (d, J=12.1 Hz, 1H), 5.27 (d, J=12.1 Hz, 1H), 3.13 (s, 4H), 2.98 (s, 3H), 2.83 (s, 3H), 2.48 (s, 3H). MS (ES+) m/z calc'd for [M+H]+[Cl2H18N4OS+H]+: 267.36 found 267.20, LCMS: tR=3.75 min [Analytical Method: 05991008_AA1.lcm]

To a mixture of 6-chloro-3-(5-nitrobenzofuran-3-yl)imidazo[1,2-b]pyridazine (12 g, 38.4 mmol) in ethanol (210 mL) and water (105 mL) was added ammonium chloride (4.11 g, 76.9 mmol) and Fe (10.73 g, 192.2 mmol). The resulting reaction mixture was stirred at 80° C. for 6 h or until the reaction was complete. Reaction mixture was filtered and concentrated to remove ethanol. Saturated aqueous sodium bicarbonate solution was added and extracted with dichloromethane (3×100 mL). Combined organic layer was dried over anhydrous sodium sulfate, filtered and concentrated to obtain crude 3-(6-chloroimidazo[1,2-b]pyridazin-3-yl)benzofuran-5-amine (1.7 g, 15.6 yield) which was used in the next step without further purification. Note: This crude material had suspected contamination of 3-Amino-6-chloropyridazine from previous steps. LCMS: RT=1.76 min; m/z=284.04, found=285.3 [M+H]+

3-Amino-6-chloropyridazine (5.7 g, 43.8 mmol) was added to a solution of 2-bromo-1-(5-nitrobenzofuran-2-yl)ethanone (12.32 g, 43.4 mmol) in DMF (350 ml) and the resulting reaction mixture was stirred at 130° C. for 16 h. Reaction mixture was cooled in an ice/water bath and diluted with ice/water to obtain solid precipitates which were filtered, washed with water and dried under high vacuum to obtain 6-chloro-3-(5-nitrobenzofuran-3-yl)imidazo[1,2-b]pyridazine (12.1 g, 89% yield).

1-(5-nitrobenzofuran-2-yl)ethanone (317 mg, 1.5 mmol) from step 1 was dissolved in dichloromethane (15 mL) and cooled under an ice/water bath. A solution of bromine (0.079 mL, 1.5 mmol) in dichloromethane (5 mL) was added drop wise at 0° C. Resulting reaction mixture was further stirred at 0° C. for 30 min and then gradually warmed up to room temperature. After 30 min, reaction mixture was quenched by the addition of saturated aqueous sodium bicarbonate solution (15 mL). Organic layer was separated and the aqueous layer was back extracted with dichloromethane (3×10 mL). Combined organic layer was washed with brine, dried over anhydrous sodium sulfate, filtered and concentrated in vacuo to give crude product 2-bromo-1-(5-nitrobenzofuran-2-yl)ethanone (400 mg, 91% yield) which was used in the next step without any further purification.

A solution of 2-hydroxy-5-nitrobenzaldehyde (500 mg, 3 mmol) and potassium hydroxide (167 mg, 3 mmol) in ethanol (8 mL) was heated at 75° C. for 5 min. Solution was gradually brought to approximately 0° C. under an ice/water bath. Chloroacetone (0.3 mL, 3.5 mmol) was added drop wise. Reaction mixture was slowly warmed up to 75° C. and stirred at this temperature for 20 h. Reaction mixture was gradually brought to room temperature and concentrated in vacuo. Water (10 mL) was added and the resulting crude mixture was extracted with ethyl acetate (3×15 mL). Combined organic layer was washed with brine (20 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. Residue was dissolved in minimum amount of ethanol by heating and subsequently cooled down to obtain precipitates. Solids were filtered off, washed with ethanol and dried under high vacuum to obtain desired product 1-(5-nitrobenzofuran-2-yl)ethanone which was used in the next step without any further purification (317 mg, 52% yield).

To a stirred solution of 6-bromo-4-chlorothieno[2,3-d]pyrimidine (50.0 g, 200.4 mmol) in anhydrous THF (2000 ml) at −78° C. was added LDA (2M, 100 mL, 200.0 mmol) dropwise. The reaction mixture was stirred at this temperature for 1 h, after which time was added H2O: THF (62.5 ml: 250 mL). The reaction mixture was warmed to 0° C. and poured into water (1000 mL), extracted with DCM (2×500 mL). The combined organic layers were dried over Na2SO4and concentrated. The crude product was triturated with hexanes to afford 5-bromo-4-chlorothieno[2,3-d]pyrimidine (32 g, 128.24 mmol, 64.1%) as light yellow solid.

Intermediate 22 is known in the literature and the synthesis can be performed according to the reference below:Reference: Bioorganic and Medicinal Chemistry Letters, 2017, vol. 27, #23, p. 5221-5224

To a stirred solution of 2,4-dichloropyrimidin mmol). The mixture was heated to 145° C. in a flask fitted with a condenser and stirred for 4 h. After cooling to room temperature, the reaction mixture was diluted with EtOAc, washed with H2O, and filtered through Celite. The filtrate was evaporated to provide crude ethyl (E)-3-(5-amino-2-chloro-pyrimidin-4-yl)acrylate (33 g) which was dissolved in 4.0 M HCl in dioxane, and the mixture was heated to 90° C. and stirred for 2 days. After cooling to room temperature, the reaction mixture was evaporated, and the residue was triturated with saturated NaHCO3solution and filtered. The cake was triturated with H2O and filtered. The product thus obtained was dried under in vacuo to provide the title compound as a brown solid (23 g, 52% over 2 steps).1H NMR (500 MHz, DMSO-d6) δ 8.72 (s, 1H), 7.83 (d, J=9.8 Hz, 1H), 7.01 (d, J=9.8 Hz, 1H). MS (ES+) m/z calcd. for [2M+H]+[2(C7H4ClN30)+H]+: 182.1 found 363.1, LCMS tR=1.63 min [Analytical Method A].

A stirred solution of 2-chloropyrido[3,2-d]pyrimidin-6(5H)-one (28.0 g, 44.1 mmol) in phosphoryl chloride (150 mL) was heated to 95° C. and stirred overnight. After cooling to room temperature, the reaction mixture was concentrated in vacuo. The resulting residue was taken up in EtOAc (500 mL) and then washed with saturated NaHCO3solution, brine, dried over Na2SO4, and concentrated in vacuo to afford 12.3 g, 42% of crude title compound as a dark brown solid which was used in the subsequent reaction without further purification.1H NMR (500 MHz, DMSO-d6) δ 9.61 (d, J=0.7 Hz, 1H), 8.49 (dd, J=8.9, 0.8 Hz, 1H), 8.14 (d, J=8.9 Hz, 1H).

Intermediate 24 is known in the literature and the synthesis can be performed according to the reference below:Reference: Bioorganic and Medicinal Chemistry, 2012, vol. 20, #15, p. 4680-4692

Available from Commercial Sources

Available from Commercial Sources

Nitrogen was bubbled through a mixture of 2-methyl-5-nitro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine (0.51 g, 1.93 mmol, 1.00 eq), 4-(2-chlorothieno[3,2-d]pyrimidin-4-yl)morpholine (0.49 g, 1.93 mmol, 1.00 eq), and Pd(PPh3)4(0.045 g, 0.0387 mmol, 0.0200 eq) in 4:1 Dioxane:2M Na2CO3(0.15 M, 13 mL) at rt for 10 minutes prior to capping the pressure vessel and lowering into an oil bath at 100° C. After 2 hours the reaction was cooled to rt and extracted with water. Organics were then concentrated in vacuo to afford 4-[2-(2-methyl-5-nitro-3-pyridyl)thieno[3,2-d]pyrimidin-4-yl]morpholine (0.68 g, 1.89 mmol) as a solid which was used directly in the next step without further purification.

Intermediate 33 is known in the literature and the synthesis can be performed according to the reference below:Reference: WO2009/26720A1

H2SO4 (0.43 M, 15 mL) was added to 3-bromo-5-hydroxy-4-methyl-benzoic acid (1.49 g, 6.44 mmol, 1.00 eq), and NaN3 (0.63 g, 9.67 mmol, 1.50 eq) in chloroform (0.32 M, 20 mL) and the resulting mixture stirred at 50° C. overnight. The reaction was cooled to rt, poured over ice, and extracted with EtOAc. The pH of the aqueous layer was then adjusted to ˜7 via the portion wise addition of sodium hydroxide pellets and then extracted with EtOAc. The second round of extractions were combined, rinsed with brine, dried over sodium sulfate, and concentrated in vacuo to provide 0.66 g, 0.871 mmol, 13.52% yield of the title compound as a dark residue used in the next reaction without further purification. 1H NMR (500 MHz, DMSO-d6) δ 11.94 (s, 1H), 9.27 (s, 1H), 6.28 (d, J=2.1 Hz, 1H), 6.08 (d, J=2.1 Hz, 1H), 2.01 (s, 3H); MS (ES+) m/z calcd. for [M+H]+[C7H8BrNO+H]+: 201.9 found 202.0 LCMS tR=3.88 min [Analytical Method 05991008_AA0.lcm].

To a stirred solution of 6-bromo-5,8-dimethyl-2-(methylsulfonyl)pyrido[2,3-d]pyrimidin-7(8H)-one (0.110 g, 0.33 mmol, 1.0 eq) in THF (10 mL), was added MeNH2in 33% ethanol (2 mL) at room temperature under nitrogen atmosphere. The reaction mixture was heated 70° C. for 16 h. The reaction mixture was evaporated under reduced pressure to afford 0.1 g, 97% yield of the title compound as a pale-yellow solid. MS (ES)+m/z calc'd for [M+H]+[C10H10BrN3O3S+H]+: 283.01, found 282.80, LCMS: tR=1.41 min [Method: C]

Synthesis of 3-bromo-2,6-difluoro-5-nitro-benzoic acid [2-43]

To a stirred solution of 3-Bromo-2,6-difluorobenzoic acid (10.00 g. 42.2 mmol. 1.00 eq.) in sulfuric acid (34.02 g, 347 mmol, 8.22 eq.) was added nitric Acid (7.98 g. 127 mmol, 3.00 eq.) drop wise at 0° C. The reaction was slowly warm to room temperature and stirred for 16 h. The reaction mixture was turned to light brown from clear solution. The reaction was poured in ice cold water (100 ml) and the obtained white precipitate was filtered through Buchner funnel and washed with water (10 ml). The solid was dried by co-distillation with toluene (3×10 mL) to afford 10 g, 83.76% yield of the title compound as an off-white solid. 1H NMR (400 MHz, DMSO-d6): δ 8.75-8.71 (m, 1H).

To a stirred solution of 3-bromo-2,6-difluoro-1-nitro-benzoic acid (5.00 g, 17.7 mmol, 1.00 eq.) in DCM (50 mL, 0.3546 M) was added DMF (0.14 mL, 1.77 mmol, 0.1000 eq.). The reaction was cooled to 0° C. and added oxalyl chloride (3.1 mL, 35.5 mmol, 2.00 eq.) dropwise. The reaction mixture was slowly warm to room temperature and stirred for 2 h. The solvent was evaporated under reduced pressure to obtain the crude acid chloride. To a stirred solution of 5-chloro-1H-pyrrolo[2,3-B]Pyridine (2.71 g, 17.7 mmol, 1.00 eq.) in DCM (30 mL) was added emim-chloride (5.20 g, 35.5 mmol, 2.00 eq.), aluminum chloride (14.19 g, 106 mmol, 6.00 eq.) and the above prepared acid chloride at 0° C. During the reaction, highly exothermic observed and the reaction mixture was turned to black color gummy liquid from clear solution. The reaction was allowed to room temperature and stirred for 16 h. The reaction mixture was basified using saturated solution of sodium bicarbonate (50 mL) and the product was extracted in Ethyl acetate (3×50 mL). The combined organic layer was dried over sodium sulphate and concentrated to give the crude product as yellow gummy solid. The crude was triturated with diethyl ether (2×25 mL) and pentane (15 mL) to afford 2.5 g, 67.69% yield of the titled compound as a yellow solid. 1H NMR (400 MHz, DMSO-d6): δ 13.48-13.11 (m, 1H), 8.86-8.74 (m, 1H), 8.55-8.51 (m, 1H), 8.51-8.49 (m, 1H), 8.47-8.44 (m, 1H); MS (ES+) m/z calc'd for [M+H]+[C14H5BrClF2N3O3+H]+: 415.92, found: 418.05, LCMS: tR=1.91 min [Method: C].

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Available from Commercial Sources

To a solution of 2,6-dichloro-phenyl-acetonitrile (30 g, 161.26 mmol 1 eq.) in dichloromethane (100 mL) and H2SO4 (80 mL) was added a mixture of H2SO4 (28.5 mL) and HNO3 (11 mL) slowly at 0° C. The reaction mixture was stirred at same temp. for 20 minutes followed by stirring at room temperature for 30 min. The progress of the reaction was monitored by TLC. After the completion, the reactant mass was concentrated to remove the organic solvent. The solution was then poured into a beaker containing ice-water (500 mL) to give a crystalline precipitate, which was filtered and washed with water to afford 20 g, 48% yield of the title compound. 1H NMR (400 MHz, CDCl3): δ 7.72 (d, J=8.8 Hz, 2H), 7.51 (d, J=8.8 Hz, 2H), 4.06 (s, 2H); MS (ES)+m/z calc'd for [M+H]+[C8H4Cl2N2O2+H]+: 230.97 found 230.65, and tR=1.814 min. LCMS: [Method: C].

To a stirred solution of ethyl 2-(2,6-dichloro-3-nitro-phenyl) acetonitrile (30 g, 129.85 mmol, 1 eq.) in Ethanol (1500 mL) was added Tin(II) chloride (81.243 g, 428.5 mmol, 3.3 eq.) in conc. HCl (300 mL) at 75° C. The reaction mixture was then stirred at room temperature for 1 h. The progress of reaction was monitored by TLC. After completion, the reaction mixture was neutralized with K2CO3to a pH of 8 and was filtered through a pad of celite bed. The filtrate was concentrated under reduced pressure to remove the solvent and dried under vacuum to get 25 g, 88.1% yield of the title compound as off white solid. 1H NMR (400 MHz, DMSO-d6): δ 7.20 (d, J=8.9 Hz, 1H), 6.83 (d, J=8.9 Hz, 1H), 5.72 (s, 2H), 4.05 (s, 2H); MS (ES)+m/z calc'd for [M+H]+[C8H6Cl2N2+H]+: 201 found 200.85, and tR=1.586 min. LCMS: [Method: C]

To a stirred solution of 2-[3-(7-amino-2-methylsulfanyl-pyrido[2,3-d]pyrimidin-6-yl)-2,4-dichloro-phenyl]isoindoline-1,3-dione (8 g, 16.586 mmol, 1.00 eq.) in TFA (73 mL) was added NaNO2(4.0055 g, 58.05 mmol, 3.5 eq.) at 0° C. under nitrogen atmosphere and the resultant reaction mixture was stirred at same temp. for another 30 mins. The reaction was monitored by TLC. After the completion of starting material, the reaction mixture was evaporated under reduced pressure and the crude was diluted with ethyl acetate and basified with Saturated K2CO3solution. The organic layer was extracted with EtOAc and the combined organic layer was dried over Na2SO4and evaporated under reduced pressure to afford the titled product 2-[2,4-dichloro-3-(2-methylsulfanyl-7-oxo-8H-pyrido[2,3-d]pyrimidin-6-yl)phenyl]isoindoline-1,3-dione. MS (ES)+m/z calc'd for [M+H]+[C22H12Cl2N4O3S+H]+: 483.01 found 482.90, and tR=1.816 min. LCMS: [Method: C].

To a stirred solution of 2-[2,4-dichloro-3-(2-methylsulfanyl-7-oxo-8H-pyrido[2,3-d]pyrimidin-6-yl)phenyl]isoindoline-1,3-dione (7.5 g, 15.517 mmol, 1 eq.) in DMF (75 mL) were added Mel (1.437 mL, 23.276 mmol, 1.5 eq.), and Cs2CO3(10.086 g. 31.035 mmol, 2 eq.). The Reaction mixture was then heated at 90° C. for 1 h. The progress of the reaction was monitored by TLC. After the completion, the reaction mixture was cooled to RT and diluted with water. The product precipitated, filtered and washed with water. The crude was further purified by chromatography by Sepaflash 40 g YMC cartridge with gradient 0-60% Ethyl acetate in n-heptane to yield 3.2 g. 27% yield of the title compound as an off-white solid. MS (ES)+m/z calc'd for [M+H]+[C23H14Cl2N4O3S+H]+:497.02 found 496.90, tR=2.085. [Method: C].

To a stirred solution of 2-[2,4-dichloro-3-(8-methyl-2-methylsulfanyl-7-oxo-pyrido[2,3-d]pyrimidin-6-yl)phenyl]isoindoline-1,3-dione (3 g, 6.032 mmol, 1 eq.) in 1,4-Dioxane (40 mL) was added Oxone (4.6295 g, 15.08 mmol, 2.5 eq.) in water (15 mL). The resultant reaction mixture was stirred at 25° C. for 18 h. The progress of reaction was monitored by TLC. After the completion, the reaction mixture was diluted with water (20 mL) and the product precipitated, filtered and washed with water to afford 2-[2,4-dichloro-3-(8-methyl-2-methylsulfonyl-7-oxo-pyrido[2,3-d] pyrimidin-6-yl) phenyl] isoindoline-1,3-dione (2.3 g mg, 72%) as a red solid. MS (ES)+m/z calc'd for [M+H]+[C23H14Cl2N4O5S+H]: 529.01 found 529, tR=1.786 mins. [Method: C].

To a stirred solution of ethyl (E)-3-(4-amino-2-methylsulfanyl-pyrimidin-5-yl)prop-2-enoate (20.00 g, 83.6 mmol, 1.00 eq.) in methanol (150 mL, 0.5572 M) was added NaOMe (4.51 g, 83.6 mmol, 1.00 eq.) at room temperature. The reaction mixture heated to 70° C. and stirred for 4 h. The reaction mixture was cooled to room temperature, concentrated under reduced pressure to obtain residue. The residue was treated with water (200 mL) and the pH was adjusted to 8 with 2N HCl. The obtained precipitate was filtered, washed with water and dried under reduced pressure to afford 15 g, 90.11% yield of the title compound as an off-white solid. MS (ES)+m/z calc'd for [M+H]+[C8H7N3OS+H]+:194.03 found 193.80, tR=1.122 min. [Method: C].

To a stirred solution of 2-methylsulfanyl-8H-pyrido[2,3-d]pyrimidin-7-one (10.00 g, 51.8 mmol, 1.00 eq.) in Acetic acid (60 mL, 0.8625 M) was added bromine (2.7 mL, 51.8 mmol, 1.0 eq.) dropwise at room temperature. The reaction mixture was heated to 70° C. and stirred for 16 h. The reaction mixture was cooled to room temperature and the precipitate was filtered, washed with DCM (50 mL), dried under reduced pressure to obtain 8 g, 56.8% yield of the crude title compound as pale yellow solid. The crude was used in the next step without further purification.1H NMR (400 MHz, DMSO-d6): δ 12.89 (br s, 1H), 8.84 (s, 1H), 8.47 (s, 1H), 2.56 (s, 3H).

To a stirred solution of 6-bromo-2-methylsulfanyl-8H-pyrido[2,3-d]pyrimidin-7-one (8.00 g, 29.4 mmol, 1.00 eq.) in DMF (100 mL, 0.2940 M) was added Cs2CO3(9.55 g, 29.4 mmol, 1.00 eq.) and Mel (1.8 mL, 29.4 mmol, 1.00 eq.) at room temperature. The reaction mixture was heated to 90° C. and stirred for 4 h. The reaction mixture was cooled to room temperature and treated with water (100 mL) to obtain precipitate. The precipitate was filtered and washed with water (30 mL), dried under reduced pressure to afford 5.0 g, 39.2% yield of the title compound as a pale yellow solid. MS (ES)+m/z calc'd for [M+H]+[C9H8BrN3OS+H]+:285.96 found 282.85. tR=1.65 min. [Method: C]

To a solution of 6-bromo-8-methyl-2-methylsulfanyl-pyrido[2,3-d]pyrimidin-7-one (5.00 g, 17.5 mmol, 1.00 eq.) in 1,4-dioxane (80 mL, 0.1664 M) was added Oxone (5.32 g, 34.9 mmol, 2.00 eq.) in Water (25 mL, 0.1664 M) at room temperature. The reaction mixture was stirred for 18 h. The reaction mixture was quenched with 1N NaHCO3solution (100 mL) and extracted with dichloromethane (3×100 mL). The combined organic layer was dried over anhydrous sodium sulphate, concentrated under reduced pressure to afford 3.5 g, 43.4% yield of the title compound as a yellow solid. MS (ES)+m/z calc'd for [M+H]+[C9H8BrN3O3S+H]+: 317.95 found 319.70, tR=1.452 mins. [Method: C].

SCA Intermediates

Available from Commercial Sources

Methyl magnesium chloride (426 mL, 1278 mmol) in anhydrous THF (852 ml) was added to a mechanically stirred 5 L round bottom flask. To this was added carbon disulfide (116 ml, 1917 mmol) in 116 ml of anhydrous THF with an addition rate such that the internal temperature did not exceed 40° C. The reaction mixture was stirred at 40° C. for 2 h. The reaction was then cooled to −78° C. using an acetone/dry ice bath, and to this was added LDA 2.0 M solution in ethyl benzene (639 ml, 1278 mmol) in 640 ml of THF. The reaction was then stirred at −78° C. for 2 h. To this was added a solution of dimethyl sulfate (243 mL, 2556 mmol,) in 246 ml of THF, and the reaction was allowed to warm to room temperature and stirred for 17 h. Then 700 ml of Et2O was added, and the reaction was stirred for 10 min and allowed to stand during which time the inorganic salts precipitated. The supernatant was decanted, and the salts were rinsed with Et2O (500 ml). The combined Et20fractions were then washed with H2O (2×1500 ml), brine (1000 ml), dried over Na2SO4, filtered, and concentrated under reduced pressure to get a dark brown oil. The crude oil was then distilled at 50° C., 3 Torr, to afford 1,1-bis(methylthio)ethylene 2 (60.00 g, 37.09%) as a yellow oil.1H NMR (500 MHz, Chloroform-d) δ 5.24 (s, 2H), 2.36 (s, 6H). LCMS: No ionization or UV signal observed.

A stirred solution of 1,1-bis(methylthio)ethylene (60.0 g, 474 mmol) in anhydrous Et2O (600 ml) was cooled to −60° C. using an Et2O/dry ice bath. To this solution was added oxalyl chloride (50 mL, 569 mmol) dropwise. During the addition of oxalyl chloride the solution turned into a yellow suspension. The suspension was allowed to warm up to −15° C., and MeOH (115 ml, 2859 mmol) was added. The reaction was then allowed to warm to room temperature and stirred for 2 h. Et2O (400 ml) was added, and the precipitate obtained was filtered and dried in vacuo to give 49.00 g, 47.60% yield of the title compound as a yellow solid.1H NMR (500 MHz, chloroform-d) δ 6.83 (s, 1H), 3.86 (s, 3H), 2.56 (s, 3H), 2.54 (s, 3H). LCMS: No ionization or UV signal observed.

To a stirred solution of 4-fluorophenylhydrazine hydrochloride (22.00 g, 135 mmol) in a dry 500 ml round-bottom flask was added THF (150 ml) and Et3N (19 ml, 135 mmol). The solution was stirred 10 min after which was added a solution of methyl 4,4-bis(methylthio)-2-oxo-but-3-enoate 4 (28.60 g, 132 mmol) in THF (150 mL). Molecular sieves (3 Å, 8-12 mesh) were added, and the reaction was heated at reflux (80° C.) for 5 h. The reaction was cooled and then filtered through Celite. The Celite was washed with EtOAc (200 ml), and the combined organic layers were evaporated in vacuo to get a crude yellow solid. This solid was further purified using ISCO, Combiflash companion, Siliasep 330 g column, (dry loaded in DCM), eluting 0 to 50% hexane/EtOAc gradient over 60 min). The product fractions were combined and concentrated in vacuo to give 13.00 g, 36.08% yield of the title compound.1H NMR (500 MHz, DMSO-d6) δ 13.00 (s, 1H), 7.64-7.57 (m, 2H), 7.44-7.37 (m, 2H), 6.92 (s, 1H), 2.47 (s, 3H). MS (ES+) m/z calcd for [M+H]+[Cl2H11FN2O2S+H]+: 252.03 found 253.0, LCMS tR=1.95 min. [Analytical Method: B]

Biochemical Assays

1. Kinase Panel

The disclosed compounds were tested for activity against a panel of at least 300 kinases. Kinase panel screening was conducted by Nanosyn (Santa Clara, CA 95051) using an enzymatic inhibition assay accepted as valid by those skilled in the art (e.g., the Caliper LabChip® mobility shift assay, an ADP detection assay, or time-resolved fluorescence detection technology. Compounds were screened at a concentration of 5 μM using an ATP concentration at the Km for each of the respective kinases and a 30-minute pre-incubation time-point.

A selection of kinases from that panel in which one or more of the disclosed compounds showed inhibition of kinase activity is shown below in Tables 1-4. In Table 1 and 2, kinase inhibition is classified by: A=95% or greater, B=90%-94%, C=80%-89%, and D=79% and less with a compound concentration of 5 JIM. In Table 3 and 4, kinase inhibition is reported in terms of IC50 values corresponding to A=less than 500 nm, B=500 nm to less than 1 μM, C=1 μM to less than 10 μM, and D=10 μM or greater using a compound concentration of 5 μM.

While we have described a number of embodiments, it is apparent that our basic examples may be altered to provide other embodiments that utilize the compounds and methods of this invention. Therefore, it will be appreciated that the scope of this invention is to be defined by the appended claims rather than by the specific embodiments that have been represented by way of example.

The contents of all references (including literature references, issued patents, published patent applications, and co-pending patent applications) cited throughout this application are hereby expressly incorporated herein in their entireties by reference. Unless otherwise defined, all technical and scientific terms used herein are accorded the meaning commonly known to one with ordinary skill in the art.