WO2009017954A1

WO2009017954A1 - Inhibitors of jak2 kinase

Info

Publication number: WO2009017954A1
Application number: PCT/US2008/069838
Authority: WO
Inventors: David Alan Campbell; Michael E. Hepperle
Original assignee: Phenomix Corporation
Priority date: 2007-08-01
Filing date: 2008-07-11
Publication date: 2009-02-05

Abstract

Compounds inhibiting the bioactivity of the protein tyrosine kinase JAK2 are provided, along with methods of using the compounds in the treatment of malconditions wherein inhibition of JAK2 is medically indicated. Methods of preparation are also provided.

Description

INHIBITORS OF JAK2 KINASE

Cross-References to Related Applications

This application claims the priority of U.S. Ser. No. 60/953,305, filed Aug. 1, 2007, which is incorporated by reference in its entirety herein.

Background The family of protein tyrosine kinases known as Janus kinases (JAK) are intracellular enzymes with at least four mammalian types, JAKl, JAK2, JAK3, and TYK2. JAK2 is associated with a wide range of cytokine receptors, including receptors activated by erythropoietin, interleukin-3, and others. An X- ray crystal structure of JAK2 at the 2.0 A resolution level in a complex with a known inhibitor has been obtained. See LS. Lucet et al. (1 January 2006), "The Structural Basis of Janus kinase 2 inhibition by a potent and specific pan- Janus kinase inhibitor," Blood, 107(1), 176-183. Among the disease states for which JAK2 inhibition may provide a therapeutic benefit are cancer and myeloproliferative conditions such as polycythemia vera. It has been found that virtually all patients with polycythemia vera, and about 50-75% of all patients with either essential thrombocythemia or de novo myelofibrosis with myeloid metaplasia, carry a mutant form of JAK2 (JAK2V617F), wherein the mutation constitutively activates JAK2. This mutant also confers cytokine hypersensitivity. Expression of this mutant in murine bone marrow transplant models results in myeloproliferative disease. See A. Pardanani, et al.

(prepublished online July 25, 2006), "MPL515 mutations in myeloproliferative and other myeloid disorders: a study of 1182 patients," Blood First Edition Paper. These data suggest that inhibition of JAK2 could provide a significant therapeutic benefit in myeloproliferative conditions and in conditions such as inflammation involving cytokines. Thus, design and preparation of inhibitors of JAK2 and other protein kinases has been studied. For example, published PCT applicationsWO2005/006156, WO2005/054230, WO2006/034116, WO2007/049041, and WO2007/013673 discuss preparation of inhibitors of protein kinases such as JAK2. Therefore there is a need for inhibitors of JAK kinase, in particular of JAK2 kinase that can be used for therapy of malconditions wherein inhibition of JAK2 activity is medically indicated.

Summary

The present invention is directed to inhibitors of JAK2 protein tyrosine kinase, methods of using the inhibitors, and methods of preparing the inhibitors.

An embodiment according to the invention concerns a compound of Formula (I):

(I) and hydrates, solvates, stereoisomers or mixtures thereof, tautomers, prodrugs, salts, crystalline forms, and mixtures thereof, wherein A, B, D, and E is each independently a carbon atom or a nitrogen atom, wherein R² is absent when A is a nitrogen atom and R³ is absent when E is a nitrogen atom, provided that least two of A, B, D, and E are nitrogen atoms and that one but not both of D and B is a nitrogen atom, and provided that neither D nor B comprises a quaternarized nitrogen atom; each dashed line represents a bond that can be present or absent, provided that bonds are present or absent so as to provide a bicyclic aromatic ring system; R¹ comprises hydrogen, alkyl, cycloalkyl, cycloalkylalkyl, aryl, aralkyl, heterocyclyl, heterocyclylalkyl, heteroaryl, heteroarylalkyl, C(O)R⁷, C(O)OR⁷, SO ₂R⁷ Or C(O)NR⁸R⁹, wherein any alkyl, aryl, aralkyl, heterocyclyl, heterocyclylalkyl, heteroaryl or heteroarylalkyl is substituted with 0-3 J groups, or, R¹ comprises -CH(R¹ ')(R¹²), wherein R¹¹ is independently hydrogen, (Ci- C₁₂)-alkyl, (C₂-C₁₂)-alkenyl, (C₂-C ₁₂)-alkynyl, (C₃-Ci₀)-cycloalkyl or (C₃-C₁₀)- cycloalkenyl, [(C₃-Ci₀)cycloalkyl or (C₃-Ci₀)-cycloalkenyl]-(C₁-Ci₂)-alkyl, (C₆- Cio)-aryl, (C₆-C_I0)-aryl-(C₁-C,₂)-alkyl, (C₃-Ci₀)-heterocyclyl, (C₃-Ci₀)- heterocyclyl-(C]-C₁₂)-alkyl, (C₅-C]₀)-heteroaryl, or (C₅-Cio)-heteroaryl-(Ci-Ci₂)- alkyl, wherein R¹ ' other than hydrogen is substituted with 0-3 substituents selected independently from J, and R¹² is independently hydrogen or (Ci-C₆)- alkyl substituted with 0-3 substituents selected independently from J; R ,R and R⁴ are independently hydrogen, alkyl, cycloalkyl, cycloalkylalkyl, aryl, aralkyl, heterocyclyl, heterocyclylalkyl, heteroaryl, heteroarylalkyl, C(O)R⁷, C(O)OR⁷, SO ₂R⁷ or C(O)NR⁸R⁹, wherein any alkyl, aryl, aralkyl, heterocyclyl, heterocyclylalkyl, heteroaryl or heteroarylalkyl is substituted with 0-3 J groups; R⁵ and R⁶ are independently halogen, CN, CF₃, OCF₃, alkyl, cycloalkyl, cycloalkylalkyl, aryl, aralkyl, heterocyclyl, heterocyclylalkyl, heteroaryl or heteroarylalkyl, wherein any alkyl, cycloalkyl, cycloalkylalkyl, aryl, aralkyl, heterocyclyl, heterocyclylalkyl, heteroaryl or heteroarylalkyl is substituted with 0-3 J; R⁷ is hydrogen, alkyl, cycloalkyl, cycloalkylalkyl, aryl, aralkyl, heterocyclyl, heterocyclylalkyl, heteroaryl or heteroarylalkyl, wherein any alkyl, cycloalkyl, cycloalkylalkyl, aryl, aralkyl, heterocyclyl, heterocyclylalkyl, heteroaryl or heteroarylalkyl is substituted with 0-3 J; X and Y are each independently present or not present, and if present are independently C(R )( R⁹), N(R¹⁰), or O, wherein R⁸ and R⁹ are independently at each occurrence hydrogen, alkyl, cycloalkyl, cycloalkylalkyl, aryl, aralkyl, heterocyclyl, heterocyclylalkyl, heteroaryl or heteroarylalkyl, wherein any alkyl, cycloalkyl, cycloalkylalkyl, aryl, aralkyl, heterocyclyl, heterocyclylalkyl, heteroaryl or heteroarylalkyl is substituted with 0-3 J, or R⁸ and R⁹, together with a carbon atom or a nitrogen atom to which they are bound, form a mono- or bicyclic ring system substituted with 0-3 J groups; R¹⁰ is hydrogen, alkyl, aryl, aralkyl, alkoxy, cycloalkyl, cycloalkylidenyl, heterocyclyl, heterocyclylalkyl, heterocyclylalkylidenyl, or heteroaryl, wherein any R¹⁰ except hydrogen is substituted with 0-3 J groups; and, J is halogen, alkyl, OR', CN, CF₃, OCF₃, R', O, S, C(O), S(O), methylenedioxy, ethylenedioxy, N(R')₂, SR', SOR', SO₂R', SO₂N(R')₂, SO₃R', C(O)R', C(O)C(O)R', C(O)CH₂C(O)R', C(S)R', C(O)OR', OC(O)R', C(0)N(R')₂, 0C(0)N(R')₂, C(S)N(R')₂, (CH₂)_0-2NHC(O)R', N(R')N(R')C(0)R', N(R')N(R')C(0)0R', N(R')N(R')CON(R')₂, N(R')S0₂R', N(R')SO₂N(R')₂, N(R')C(0)0R', N(R')C(O)R', N(R')C(S)R', N(R')C(0)N(R')₂, N(R')C(S)N(R')₂, N(C0R')C0R', N(OR')R', C(=NH)N(R')₂, C(0)N(0R')R^τ, or C(=N0R')R' wherein each R' is independently at each occurrence hydrogen, (Cp C₁₂)-alkyl, (C₂-C₁₂)alkenyl, (C₂-C₁₂)alkynyl, (C₃-C₁₀)-cycloalkyl, (C₃-C₁₀)- cycloalkenyl, [(C₃-C i₀)cycloalkyl, (C₃-Ci₀)-cycloalkenyl]-(Ci-C,₂)-alkyl, (C₆- Cio)-aryl, (C₆-Ci₀)-aryl-(C,-C,₂)-alkyl, (C₃-C i₀)-heterocyclyl, (C₃-C₁₀)- heterocyclyl-(Ci-Ci₂)-alkyl, (C₅-C i₀)-heteroaryl, or (C₅-Ci₀)-heteroaryl-(Ci-Ci₂)- alkyl, wherein R' is substituted with 0-3 substituents selected independently from J, or, when two R' are bound to a nitrogen atom or to two adjacent nitrogen atoms, the two R' groups together with the nitrogen atom or atoms to which they are bound can form a 3- to 8-membered monocyclic ring, or an 8- to 20- membered bi- or tricyclic heterocyclic ring system, wherein any ring can further contain 1-3 additional heteroatoms selected from the group consisting of N, NR', NH, O, S, SO and SO₂ and wherein each ring is substituted with 0-3 substituents selected independently from J; wherein, in the bi- and tricyclic ring system, each ring is linearly fused, bridged, or spirocyclic, wherein each ring is either aromatic or nonaromatic, wherein each ring can be fused to a (C₆-C io)aryl, (C₅- Cio)heteroaryl, (C₃-C io)cycloalkyl or (C₃-Ci₀)heterocyclyl. Another embodiment according to the invention concerns a method of treating a malcondition in a patient wherein inhibition of JAK2 is medically indicated, comprising providing a compound of any one of claims 1-15 to the patient in a dosage, at a frequency, and for a duration sufficient to provide a beneficial effect to the patient. The JAK2 can be a mutant form of the enzyme. The malcondition can include cancer or a myeloproliferative disease, an autoimmune disorder, or a malcondition wherein modulation of cytokine signaling by inhibition of JAK2 is medically indicated.

Another embodiment of the invention is directed to a pharmaceutical composition comprising an inventive compound and a medically acceptable excipient.

Another embodiment of the invention is directed to a pharmaceutical combination comprising a compound of the invention and a second medicament.

Another embodiment of the invention is directed to a method of preparation of inventive pyrazolopyrimidines. Another embodiment of the invention is directed to a method of preparation of inventive triazolopyridines. Detailed Description of the Invention

Definitions

"Patient" as used herein, includes mammals such as humans, non-human primates, rats, mice, dogs, cats, horses, cows and pigs. The terms "treatment," "treating," therapy," and "therapeutic" are defined as relating to the management and care of a patient for the purpose of combating the disease, condition, or disorder and includes administering a compound of the present invention to prevent the onset of the symptoms or complications, or alleviating the symptoms or complications, or eliminating the disease, condition, or disorder.

"Treating" within the context of the instant invention means an alleviation of symptoms associated with a disorder or disease, or inhibition of further progression or worsening of those symptoms, or prevention or prophylaxis of the disease or disorder. Thus, treating a myeloproliferative condition includes the control, alleviation or prevention of symptoms of the condition. Similarly, as used herein, an "effective amount" or a "therapeutically effective amount" of a compound of the invention refers to an amount of the compound that alleviates, in whole or in part, symptoms associated with the disorder or condition, or halts or slows further progression or worsening of those symptoms, or prevents or provides prophylaxis for the disorder or condition or increases the survival of the patient. In particular, a dose, frequency, and duration sufficient to provide a beneficial result refers to an amount effective, at dosages and for periods of time necessary, to achieve the desired therapeutic result by inhibition of JAK2 activity. A therapeutically effective amount is also one in which any toxic or detrimental effects of compounds of the invention are outweighed by the therapeutically beneficial effects.

All chiral, diastereomeric, racemic forms of a structure are intended, unless the specific stereochemistry or isomeric form is specifically indicated. Compounds used in the present invention include enriched or resolved optical isomers at any or all asymmetric atoms as are apparent from the depictions. Both racemic and diastereomeric mixtures, as well as the individual optical isomers can be isolated or synthesized so as to be highly enriched in their enantiomeric or diastereomeric partners, and these are all within the scope of the invention. In general, "substituted" refers to an organic group as defined herein in which one or more bonds to a hydrogen atom contained therein are replaced by a bond to non-hydrogen or non-carbon atoms such as, but not limited to, a halogen (i.e., F, Cl, Br, and I); an oxygen atom in groups such as hydroxyl groups, alkoxy groups, aryloxy groups, aralkyloxy groups; a sulfur atom in groups such as thiol groups, alkyl and aryl sulfide groups, sulfoxide groups, sulfone groups, sulfonyl groups, and sulfonamide groups; a nitrogen atom in groups such as amines, hydroxylamines, N-oxides, hydrazides, azides, and enamines; and other heteroatoms in various other groups. Substituted alkyl, alkenyl, alkynyl, cycloalkyl, and cycloalkenyl groups as well as other substituted groups also include groups in which one or more bonds to a carbon(s) or hydrogen(s) atom are replaced by one or more bonds, including double or triple bonds, to a heteroatom such as, but not limited to, oxygen in carbonyl (oxo), carboxyl, ester, amide, imide, urethane, and urea groups; and nitrogen in imines, hydroxyimines, oximes, hydrazones, amidines, guanidines, and nitriles.

The term "heteroatoms" as used herein refers to non-carbon and non- hydrogen atoms, and is not otherwise limited. Typical heteroatoms are N, O, and S. When sulfur (S) is referred to, it is understood that the sulfur can be in any of the oxidation states in which it is found, thus including sulfoxides (R- S(O)-R') and sulfones (R-S(O)₂-R'), unless the oxidation state is specified; thus, the term "sulfone" encompasses only the sulfone form of sulfur; the term "sulfide" encompasses only the sulfide (R-S-R') form of sulfur. When the phrases such as "heteroatoms selected from the group consisting of O, NH, NR' and S," or "[variable] is O, S . . ." are used, they are understood to encompass all of the sulfide, sulfoxide and sulfone oxidation states of sulfur.

Substituted ring groups such as substituted aryl, heterocyclyl and heteroaryl groups also include rings and fused ring systems in which a bond to a hydrogen atom is replaced with a bond to a carbon atom. Therefore, substituted aryl, heterocyclyl and heteroaryl groups may also be substituted with alkyl, alkenyl, and alkynyl groups as defined herein.

Alkyl groups include straight chain and branched alkyl groups and cycloalkyl groups having from 1 to about 20 carbon atoms, and typically from 1 to 12 carbons or, in some embodiments, from 1 to 8 carbon atoms. Examples of straight chain alkyl groups include those with from 1 to 8 carbon atoms such as methyl, ethyl, n-propyl, n-butyl, n-pentyl, n-hexyl, n-heptyl, and n-octyl groups. Examples of branched alkyl groups include, but are not limited to, isopropyl, iso-butyl, sec-butyl, t-butyl, neopentyl, isopentyl, and 2,2-dimethylpropyl groups. Representative substituted alkyl groups may be substituted one or more times with any of the groups listed above, for example, amino, hydroxy, cyano, carboxy, nitro, thio, alkoxy, and halogen groups.

Cycloalkyl groups are cyclic alkyl groups such as, but not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl groups. In some embodiments, the cycloalkyl group has 3 to 8 ring members, whereas in other embodiments the number of ring carbon atoms range from 3 to 5, 6, or 7. Cycloalkyl groups further include polycyclic cycloalkyl groups such as, but not limited to, norbornyl, adamantyl, bornyl, camphenyl, isocamphenyl, and carenyl groups, and fused rings such as, but not limited to, decalinyl, and the like. Cycloalkyl groups also include rings that are substituted with straight or branched chain alkyl groups as defined above. Representative substituted cycloalkyl groups may be mono-substituted or substituted more than once, such as, but not limited to, 2,2-, 2,3-, 2,4- 2,5- or 2,6-disubstituted cyclohexyl groups or mono-, di- or tri-substituted norbornyl or cycloheptyl groups, which may be substituted with, for example, amino, hydroxy, cyano, carboxy, nitro, thio, alkoxy, and halogen groups.

(Cycloalkyl)alkyl groups, also denoted cycloalkylalkyl, are alkyl groups as defined above in which a hydrogen or carbon bond of the alkyl group is replaced with a bond to a cycloalkyl group as defined above.

Alkenyl groups include straight and branched chain and cyclic alkyl groups as defined above, except that at least one double bond exists between two carbon atoms. Thus, alkenyl groups have from 2 to about 20 carbon atoms, and typically from 2 to 12 carbons or, in some embodiments, from 2 to 8 carbon atoms. Examples include, but are not limited to vinyl, -CH=CH(CH₃), -CH=C(CH₃)₂, -C(CH₃)=CH₂, -C(CH₃)=CH(CH₃), -C(CH₂CH₃)^CH₂, cyclohexenyl, cyclopentenyl, cyclohexadienyl, butadienyl, pentadienyl, and hexadienyl among others.

Cycloalkenyl groups include cycloalkyl groups having at least one double bond between 2 carbons. Thus for example, cycloalkenyl groups include but are not limited to cyclohexenyl, cyclopentenyl, cyclohexadienyl, cyclobutenyl, cyclopentadienyl, and cyclohexadienyl groups.

(Cycloalkenyl)alkyl groups are alkyl groups as defined above in which a hydrogen or carbon bond of the alkyl group is replaced with a bond to a cycloalkenyl group as defined above.

Alkynyl groups include straight and branched chain alkyl groups, except that at least one triple bond exists between two carbon atoms. Thus, alkynyl groups have from 2 to about 20 carbon atoms, and typically from 2 to 12 carbons or, in some embodiments, from 2 to 8 carbon atoms. Examples include, but are not limited to -C≡CH, -C=C(CH₃), -C≡C(CH₂CH₃), -CH₂C≡CH, -CH₂C≡C(CH₃), and -CH₂C=C(CH₂CH₃) among others.

Aryl groups are cyclic aromatic hydrocarbons that do not contain heteroatoms. Thus aryl groups include, but are not limited to, phenyl, azulenyl, heptalenyl, biphenyl, indacenyl, fluorenyl, phenanthrenyl, triphenylenyl, pyrenyl, naphthacenyl, chrysenyl, biphenylenyl, anthracenyl, and naphthyl groups. In some embodiments, aryl groups contain 6-14 carbons in the ring portions of the groups. Although the phrase "aryl groups" includes groups containing fused rings, such as fused aromatic-aliphatic ring systems (e.g., indanyl, tetrahydronaphthyl, and the like), it does not include aryl groups that have other groups, such as alkyl or halogen groups, bonded to one of the ring members. Rather, groups such as tolyl are referred to as substituted aryl groups. Representative substituted aryl groups may be mono-substituted or substituted more than once, such as, but not limited to, 2-, 3-, 4-, 5-, or 6-substituted phenyl or naphthyl groups, which may be substituted with groups such as those listed above.

Aralkyl groups are alkyl groups as defined above in which a hydrogen or carbon bond of an alkyl group is replaced with a bond to an aryl group as defined above. Representative aralkyl groups include benzyl and phenylethyl groups and fused (cycloalkylaryl)alkyl groups such as 4-ethyl-indanyl. Aralkenyl group are alkenyl groups as defined above in which a hydrogen or carbon bond of an alkyl group is replaced with a bond to an aryl group as defined above.

Heterocyclyl groups include aromatic and non-aromatic ring compounds containing 3 or more ring members, of which, one or more is a heteroatom such as, but not limited to, N, O, and S. In some embodiments, heterocyclyl groups include 3 to 20 ring members, whereas other such groups have 3 to 15 ring members. The phrase "heterocyclyl group" includes fused ring species including those comprising fused aromatic and non-aromatic groups. The phrase also includes polycyclic ring systems containing a heteroatom such as, but not limited to, quinuclidyl. However, the phrase does not include heterocyclyl groups that have other groups, such as alkyl or halogen groups, bonded to one of the ring members. Rather, these are referred to as "substituted heterocyclyl groups". Heterocyclyl groups include, but are not limited to, pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, pyrrolyl, pyrazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, thiazolyl, pyridinyl, thiophenyl, benzothiophenyl, benzofuranyl, dihydrobenzofuranyl, indolyl, dihydroindolyl, azaindolyl, indazolyl, benzimidazolyl, azabenzimidazolyl, benzoxazolyl, benzothiazolyl, benzothiadiazolyl, imidazopyridinyl, isoxazolopyridinyl, thianaphthalenyl, purinyl, xanthinyl, adeninyl, guaninyl, quinolinyl, isoquinolinyl, tetrahydroquinolinyl, quinoxalinyl, and quinazolinyl groups. Representative substituted heterocyclyl groups may be mono-substituted or substituted more than once, such as, but not limited to, piperidinyl or quinolinyl groups, which are 2-, 3-, 4-, 5-, or 6-substituted, or disubstituted with groups such as those listed above.

Heteroaryl groups are aromatic ring compounds containing 5 or more ring members, of which, one or more is a heteroatom such as, but not limited to, N, O, and S. Heteroaryl groups include, but are not limited to, groups such as pyrrolyl, pyrazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, thiazolyl, pyridinyl, thiophenyl, benzothiophenyl, benzofuranyl, indolyl, azaindolyl, indazolyl, benzimidazolyl, azabenzimidazolyl, benzoxazolyl, benzothiazolyl, benzothiadiazolyl, imidazopyridinyl, isoxazolopyridinyl, thianaphthalenyl, purinyl, xanthinyl, adeninyl, guaninyl, quinolinyl, isoquinolinyl, tetrahydroquinolinyl, quinoxalinyl, and quinazolinyl groups. Although the phrase "heteroaryl groups" includes fused ring compounds such as indolyl, isoindolyl, and 2,3-dihydro indolyl, the phrase does not include heteroaryl groups that have other groups bonded to one of the ring members, such as alkyl groups. Rather, heteroaryl groups with such substitution are referred to as "substituted heteroaryl groups." Representative substituted heteroaryl groups may be substituted one or more times with groups such as those listed above. Additional examples of aryl and heteroaryl groups include but are not limited to phenyl, biphenyl, indenyl, naphthyl (1-naphthyl, 2-naphthyl), N- hydroxytetrazolyl, N-hydroxytriazolyl, N-hydroxyimidazolyl, anthracenyl (1- anthracenyl, 2-anthracenyl, 3 -anthracenyl), thiophenyl (2-thienyl, 3-thienyl), furyl (2-furyl, 3-furyl) , indolyl, oxadiazolyl, isoxazolyl, quinazolinyl, fluorenyl, xanthenyl, isoindanyl, benzhydryl, acridinyl, thiazolyl, pyrrolyl (2-pyrrolyl), pyrazolyl (3-pyrazolyl), imidazolyl (1-imidazolyl, 2-imidazolyl, 4-imidazolyl, 5- imidazolyl), triazolyl (1,2,3-triazol-l-yl, l,2,3-triazol-2-yl l,2,3-triazol-4-yl, l,2,4-triazol-3-yl), oxazolyl (2-oxazolyl, 4-oxazolyl, 5-oxazolyl), thiazolyl (2- thiazolyl, 4-thiazolyl, 5-thiazolyl), pyridyl (2-pyridyl, 3-pyridyl, 4-pyridyl), pyrimidinyl (2-pyrimidinyl, 4-pyrimidinyl, 5-pyrimidinyl, 6-pyrimidinyl), pyrazinyl, pyridazinyl (3- pyridazinyl, 4-pyridazinyl, 5-pyridazinyl), quinolyl (2- quinolyl, 3-quinolyl, 4-quinolyl, 5-quinolyl, 6-quinolyl, 7-quinolyl, 8-quinolyl), isoquinolyl (1-isoquinolyl, 3-isoquinolyl, 4-isoquinolyl, 5-isoquinolyl, 6- isoquinolyl, 7-isoquinolyl, 8-isoquinolyl), benzo[b]furanyl (2-benzo[b]furanyl, 3-benzo[b]furanyl, 4-benzo[b]furanyl, 5-benzo[b]furanyl, 6-benzo[b]furanyl, 7- benzo[b]furanyl), 2,3-dihydro-benzo[b]furanyl (2-(2,3-dihydro- benzo[b]furanyl), 3-(2,3-dihydro-benzo[b]furanyl), 4-(2,3-dihydro- benzo[b]furanyl), 5-(2,3-dihydro-benzo[b]furanyl), 6-(2,3-dihydro- benzo[b]furanyl), 7-(2,3-dihydro-benzo[b]furanyl), benzo[b] thiophenyl (2- benzo[b]thiophenyl, 3-benzo[b]thiophenyl, 4-benzo[b]thiophenyl, 5-benzo[b]thiophenyl, 6-benzo[b]thiophenyl, 7-benzo[b]thiophenyl), 2,3-dihydro-benzo[b]thiophenyl, (2-(2,3-dihydro-benzo[b]thiophenyl), 3-(2,3- dihydro-benzo[b]thiophenyl), 4-(2,3-dihydro-benzo[b]thiophenyl), 5-(2,3- dihydro-benzo[b]thiophenyl), 6-(2,3-dihydro-benzo[b]thiophenyl), 7-(2,3- dihydro-benzo[b]thiophenyl), indolyl (1 -indolyl, 2-indolyl, 3 -indolyl, 4-indolyl, 5-indolyl, 6-indolyl, 7-indolyl), indazole (1-indazolyl, 3-indazolyl, 4-indazolyl, 5-indazolyl, 6-indazolyl, 7-indazolyl), benzimidazolyl (1-benzimidazolyl, 2-benzimidazolyl, 4-benzimidazolyl, 5 -benzimidazolyl, 6-benzimidazolyl, 7-benzimidazolyl, 8-benzimidazolyl), benzoxazolyl (1-benzoxazolyl, 2- benzoxazolyl), benzothiazolyl (1-benzothiazolyl, 2-benzothiazolyl, A- benzothiazolyl, 5 -benzothiazolyl, 6-benzothiazolyl, 7-benzothiazolyl), carbazolyl (1-carbazolyl, 2-carbazolyl, 3-carbazolyl, 4-carbazolyl), 5H-dibenz[b,f]azepine (5H-dibenz[b,f]azepin-l-yl, 5H-dibenz[b,f]azepine-2-yl, 5H-dibenz[b,f]azepine-3-yl, 5H-dibenz[b,f]azepine-4-yl, 5H-dibenz[b,f]azepine- 5-yl), 10,l l-dihydro-5H-dibenz[b,f]azepine (10,l l-dihydro-5H- dibenz[b,f]azepine-l-yl, 10,l l-dihydro-5H-dibenz[b,f]azepine-2-yl, 10,11- dihydro-5H-dibenz[b,f]azepine-3-yl, 10,l l-dihydro-5H-dibenz[b,f]azepine-4-yl, 10,1 l-dihydro-5H-dibenz[b,f]azepine-5-yl), and the like.

Heterocyclylalkyl groups are alkyl groups as defined above in which a hydrogen or carbon bond of an alkyl group is replaced with a bond to a heterocyclyl group as defined above. Representative heterocyclyl alkyl groups include, but are not limited to, furan-2-yl methyl, furan-3-yl methyl, pyridine-3- yl methyl, tetrahydrofuran-2-yl ethyl, and indol-2-yl propyl.

Heteroarylalkyl groups are alkyl groups as defined above in which a hydrogen or carbon bond of an alkyl group is replaced with a bond to a heteroaryl group as defined above.

The term "alkoxy" refers to an oxygen atom connected to an alkyl group as defined above. Examples of linear alkoxy groups include but are not limited to methoxy, ethoxy, propoxy, butoxy, pentyloxy, hexyloxy, and the like. Examples of branched alkoxy include but are not limited to isopropoxy, sec- butoxy, tert-butoxy, isopentyloxy, isohexyloxy, and the like. Examples of cyclic alkoxy include but are not limited to cyclopropyloxy, cyclobutyloxy, cyclopentyloxy, cyclohexyloxy, and the like.

The terms "aryloxy" and "arylalkoxy" refer to, respectively, an aryl group bonded to an oxygen atom and an aralkyl group bonded to the oxygen atom at the alkyl. Examples include but are not limited to phenoxy, naphthyloxy, and benzyloxy.

The term "amine" (or "amino") includes primary, secondary, and tertiary amines having, e.g., the formula -NR₂. Amines include but are not limited to -NH₂, alkylamines, dialkylamines, arylamines, alkylarylamines, diarylamines, aralkylamines, heterocyclylamines and the like.

The term "amide" (or "amido") includes C- and N-amide groups, i.e., -C(O)NR₂, and -NRC(O)R groups, respectively. Amide groups therefore include but are not limited to carbamoyl groups (-C(O)NH₂) and formamide groups (-NHC(O)H). The term "urethane" (or "carbamyl") includes N- and O-urethane groups, i.e., -NRC(O)OR and -OC(O)NR₂ groups, respectively.

The term "sulfonamide" (or "sulfonamido") includes S- and N- sulfonamide groups, i.e., -SO₂NR₂ and -NRSO₂R groups, respectively. Sulfonamide groups therefore include but are not limited to sulfamoyl groups (- SO₂NH₂).

A "core ring system" as the term is used herein refers to the bicyclic aromatic system defined as a compound of the formula:

wherein the atoms A, B, D, and E include two or three nitrogen atoms, the remaining atoms being carbon, and provided that only one of B and D is a nitrogen and the other is carbon, and the nitrogen is not a quaternarized nitrogen. The dashed lines represent bonds being present or absent, that is, having single or double bonds at those positions, to provide a bicyclic aromatic ring system wherein double bonds are conjugated, including through a nitrogen atom (for example, as pyrrole or pyridine).

In addition, where features or aspects of the invention are described in terms of Markush groups, those skilled in the art will recognize that the invention is also thereby described in terms of any individual member or subgroup of members of the Markush group. For example, if A is described as selected from the group consisting of bromine, chlorine, and iodine, claims for A being bromine and claims for A being bromine and chlorine are fully described. Moreover, where features or aspects of the invention are described in terms of Markush groups, those skilled in the art will recognize that the invention is also thereby described in terms of any combination of individual members or subgroups of members of Markush groups. Thus, for example, if A is described as selected from the group consisting of bromine, chlorine, and iodine, and B is described as selected from the group consisting of methyl, ethyl, and propyl, claims for A being bromine and B being methyl are fully described. Detailed Description

An embodiment according to the present invention is directed to a compound of Formula (I),

(I) wherein the substituent groups are as defined herein. The inventive compounds can inhibit JAK2, and thus can be useful for treatment of malconditions in patients, such as human beings, where JAK2 inhibition is medically indicated. Atoms A, B, D, and E can be either nitrogen atoms or carbon atoms in any arrangement, provided that at least two of A, B, D, and E are nitrogen atoms (thus providing a core ring system with at least three nitrogen atoms including the 5-ring nitrogen atom explicitly indicated), and also provided that one of the ring junction atoms B and D is a nitrogen atom and the other a carbon atom. Thus the core ring system of the inventive compounds can include up to a total of 4 nitrogen atoms, including the 5-ring nitrogen atom explicitly indicated. No ring nitrogen is a quaternary salt, and double and single bonds are disposed in each structure accordingly. An atom A or E can bear a substituent R² or R³ respectively when the atom is a carbon atom, but when the atom is nitrogen instead of carbon, the respective substituent is absent. For example, atoms B and E can both be nitrogen atoms and A and D be carbon atoms, providing a triazolopyridine ring core of the formula (XI):

(XI) wherein R³ is absent, and X, Y, R¹, R², R⁴, R⁵ and R⁶ are as defined herein. Alternatively, atoms A and D can both be nitrogen atoms, and B and E both be carbon atoms, providing a pyrazolopyrimidine core of the formula (XII):

(XII) wherein R² is absent, and X, Y, R¹, R³, R⁴, R⁵ and R⁶ are as defined herein.

Other core ring systems containing a total of three nitrogen atoms are also within the invention, such as the system where A and B are nitrogen, and D and E are carbon, formula (XIII):

(XIII), providing an imidazolopyridazine, or the system wherein D and E are nitrogen and A and B are carbon, formula (XIV):

(XIV), providing an isomeric triazolopyridine.

Alternatively, the inventive compound can include a ring core wherein there are a total of four nitrogen atoms. For example an inventive compound can have a ring core of formula (XV):

(XV) providing a triazolopyridazine. Or, an inventive compound can have a ring core of the formula (XVI):

(XVI) providing a triazolopyrimidine.

The six structures of formula (XI)-(XVI) show the six theoretical core ring systems for the compounds of the invention.

Alternatively, the inventive compound can comprise a compound of formula (II)

providing a pyrazolopyrimidine ring core wherein the substituents are as defined herein.

In the compound of formula (I), the linker atoms X and Y can each be independently present or not present, and if present can be independently a carbon atom, optionally substituted with R⁸ and R⁹; a nitrogen atom, optionally substituted with R¹⁰' or an oxygen atom. When X or Y is absent, the corresponding substituent of formula (I), R⁴ or R¹ respectively, is bonded directly to the ring. Either R⁴, R¹, or both, can be hydrogen, so in an embodiment of the inventive compound, one or both of these ring positions can be unsubstituted (that is, bearing a hydrogen atom).

In an embodiment of the invention, an inventive compound includes those compounds of formula (I) wherein at least one of X and Y comprises NR¹⁰, R¹⁰ comprising hydrogen, alkyl, aryl, aralkyl, alkoxy, cycloalkyl, cycloalkylidenyl, heterocyclyl, heterocyclylalkyl, heterocyclylalkylidenyl, or heteroaryl, wherein any R¹⁰ except hydrogen is substituted with 0-3 J groups, J being a mono- or a divalent substituent radical as is defined herein. The group X, when equivalent to NR¹⁰, is also substituted with R⁴ which can be hydrogen, alkyl, cycloalkyl, cycloalkylalkyl, aryl, aralkyl, heterocyclyl, heterocyclylalkyl, heteroaryl, heteroarylalkyl, C(O)R⁷, C(O)OR⁷, SO ₂R⁷ Or C(O)NR⁸R⁹; thus the core ring system is an amino substituted aromatic ring system, an amido substitued ring system, a urethane substituted ring system, a urea substituted ring system, or a sulfonamido substituted ring system.

For examples of an inventive compound wherein X is nitrogen, see examples 15-18 in Table 1. When X is NR¹⁰, Y can be absent, and R¹ can be hydrogen, to provide a core ring system bearing a hydrogen on the ring carbon bonded to Y. More specifically, R⁴, bonded to X, can be C(O)R⁷, and R¹⁰ can be H, providing a core ring system substituted with a R⁷C(O)NH- group on the fϊve-membered ring of the core ring system. More specifically, R⁷ can be alkyl or cycloalkyl, such as isopropyl, cyclobutyl, or cyclopentyl.

More specifically, an inventive compound can include an aryl group at position R². For example, the above substitution pattern wherein Y-R¹ is hydrogen, and atom A is a carbon atom, can include an aryl group bonded to that carbon atom. For example, the aryl group can be a m-chlorophenyl group or a 2- methoxy-5-isopropylphenyl group.

The group Y, when equivalent to NR¹⁰, is also substituted with R¹, which can be hydrogen, alkyl, cycloalkyl, cycloalkylalkyl, aryl, aralkyl, heterocyclyl, heterocyclylalkyl, heteroaryl, heteroarylalkyl, C(O)R⁷, C(O)OR⁷, SO₂R⁷ Or C(O)NR⁸R⁹, wherein any alkyl, aryl, aralkyl, heterocyclyl, heterocyclylalkyl, heteroaryl or heteroarylalkyl is substituted with 0-3 J groups, or, R¹ can be a substituted methine group bearing R¹ ' and R¹². In these embodiments, the core ring system is also an amino substituted aromatic ring system, an amido substituted ring system, a urethane substituted ring system, a urea substituted ring system, or a sulfonamido substituted ring system. For examples of an inventive compound wherein Y is nitrogen, see examples 5-10 in Table 1.

When Y is NR¹⁰R¹, and X is absent and R⁴ is hydrogen, to provide a core ring system bearing a hydrogen on the ring carbon bonded to X, R¹⁰ can be hydrogen. R¹ can comprise an aryl, aralkyl, heteroaryl, or heteroarylalkyl group. For example, R¹ can be a p-fluoro-α-phenethyl group. Alternatively, R¹ can be a α-pyridylethyl group, such as a 2-pyridylethyl or 3-pyridylethyl group, bonded to X via the α-carbon. These groups include a chiral carbon, that is, the groups exist in stereoisomeric forms. Accordingly, the inventive compounds bearing α- phenethyl or α-pyridylethyl groups can be racemic mixtures at that chiral center, or can be a pure single enantiomer, or can be mixtures thereof.

When both X and Y are equivalent to NR¹⁰, bearing respectively the R⁴ and R¹ groups, the core ring system can be any of the combinations of the amino, amide, urethane, urea, or sulfonamido substituent groups, for examine, the inventive compound can include a diamino substituted core ring system, or can include an amino-amido disubstituted core ring system, or any other combination. Specifically, the inventive compounds include any of the six specific core ring systems illustrated above of formulas (XI)-(XVI) bearing any of these combinations of X and Y wherein one or both comprise NR¹⁰ with its respective R⁴ and/or R¹ group.

R² and R³, when present, can be aryl groups bonded to a ring carbon. For example, they can be halogenated aryl groups, such as chlorophenyl or flurophenyl groups, or can bear other substituents, such as alkyl and alkoxy groups. For example, R² or R³ can be a 2-methoxy-5-isopropylphenyl group.

Specific examples of inventive compounds are provided below in Table 1.

Compositions and Combination Treatments A. Compositions.

Another aspect of the invention provides compositions of the compounds of the invention, alone or in combination with another JAK2 inhibitor or another type of therapeutic agent. As set forth herein, compounds of the invention include stereoisomers, tautomers, solvates, prodrugs, pharmaceutically acceptable salts and mixtures thereof. Compositions containing a compound of the invention may be prepared by conventional techniques, e.g. as described in Remington: The Science and Practice of Pharmacy, 21st Ed., (2005). The compositions may appear in conventional forms, for example capsules, tablets, aerosols, solutions, suspensions or topical applications. Typical compositions include a compound of the invention which inhibits the enzymatic activity of JAK2 kinase, and a pharmaceutically acceptable excipient which may be a carrier or a diluent. For example, the active compound will usually be mixed with a carrier, or diluted by a carrier, or enclosed within a carrier which may be in the form of an ampoule, capsule, sachet, paper, or other container. When the active compound is mixed with a carrier, or when the carrier serves as a diluent, it may be solid, semi-solid, or liquid material that acts as a vehicle, excipient, or medium for the active compound. The active compound can be adsorbed on a granular solid carrier, for example contained in a sachet. Some examples of suitable carriers are water, salt solutions, alcohols, polyethylene glycols, polyhydroxyethoxylated castor oil, peanut oil, olive oil, gelatin, lactose, terra alba, sucrose, dextrin, magnesium carbonate, sugar, cyclodextrin, amylose, magnesium stearate, talc, gelatin, agar, pectin, acacia, stearic acid or lower alkyl ethers of cellulose, silicic acid, fatty acids, fatty acid amines, fatty acid monoglycerides and diglycerides, pentaerythritol fatty acid esters, polyoxyethylene, hydroxymethylcellulose and polyvinylpyrrolidone. Similarly, the carrier or diluent may include any sustained release material known in the art, such as glyceryl monostearate or glyceryl distearate, alone or mixed with a wax. The formulations can be mixed with auxiliary agents which do not deleteriously react with the active compounds. Such additives can include wetting agents, emulsifying and suspending agents, salt for influencing osmotic pressure, buffers and/or coloring substances preserving agents, sweetening agents or flavoring agents. The compositions can also be sterilized if desired. The route of administration may be any route which effectively transports the active compound of the invention which inhibits the enzymatic activity of the JAK2 kinase to the appropriate or desired site of action, such as oral, nasal, pulmonary, buccal, subdermal, intradermal, transdermal or parenteral, e.g., rectal, depot, subcutaneous, intravenous, intraurethral, intramuscular, intranasal, ophthalmic solution or an ointment, the oral route being preferred.

If a solid carrier is used for oral administration, the preparation may be tabletted, placed in a hard gelatin capsule in powder or pellet form or it can be in the form of a troche or lozenge. If a liquid carrier is used, the preparation may be in the form of a syrup, emulsion, soft gelatin capsule or sterile injectable liquid such as an aqueous or non-aqueous liquid suspension or solution.

Injectable dosage forms generally include aqueous suspensions or oil suspensions which may be prepared using a suitable dispersant or wetting agent and a suspending agent Injectable forms may be in solution phase or in the form of a suspension, which is prepared with a solvent or diluent. Acceptable solvents or vehicles include sterilized water, Ringer's solution, or an isotonic aqueous saline solution. Alternatively, sterile oils may be employed as solvents or suspending agents. Preferably, the oil or fatty acid is non-volatile, including natural or synthetic oils, fatty acids, mono-, di- or tri-glycerides.

For injection, the formulation may also be a powder suitable for reconstitution with an appropriate solution as described above. Examples of these include, but are not limited to, freeze dried, rotary dried or spray dried powders, amorphous powders, granules, precipitates, or particulates. For injection, the formulations may optionally contain stabilizers, pH modifiers, surfactants, bioavailability modifiers and combinations of these. The compounds may be formulated for parenteral administration by injection such as by bolus injection or continuous infusion. A unit dosage form for injection may be in ampoules or in multi-dose containers. The formulations of the invention may be designed to provide quick, sustained, or delayed release of the active ingredient after administration to the patient by employing procedures well known in the art. Thus, the formulations may also be formulated for controlled release or for slow release.

Compositions contemplated by the present invention may comprise, for example, micelles or liposomes, or some other encapsulated form, or may be administered in an extended release form to provide a prolonged storage and/or delivery effect. Therefore, the formulations may be compressed into pellets or cylinders and implanted intramuscularly or subcutaneously as depot injections or as implants such as stents. Such implants may employ known inert materials such as silicones and biodegradable polymers, e.g., polylactide-polyglycolide. Examples of other biodegradable polymers include poly(orthoesters) and poly(anhydrides) .

For nasal administration, the preparation may contain a compound of the invention which inhibits the enzymatic activity of the JAK2 kinase, dissolved or suspended in a liquid carrier, preferably an aqueous carrier, for aerosol application. The carrier may contain additives such as solubilizing agents, e.g., propylene glycol, surfactants, absorption enhancers such as lecithin (phosphatidylcholine) or cyclodextrin, or preservatives such as parabens. For parenteral application, particularly suitable are injectable solutions or suspensions, preferably aqueous solutions with the active compound dissolved in polyhydroxylated castor oil.

Tablets, dragees, or capsules having talc and/or a carbohydrate carrier or binder or the like are particularly suitable for oral application. Preferable carriers for tablets, dragees, or capsules include lactose, corn starch, and/or potato starch. A syrup or elixir can be used in cases where a sweetened vehicle can be employed.

A typical tablet that may be prepared by conventional tabletting techniques may contain:

Core:

Active compound (as free compound or salt thereof) 250 mg Colloidal silicon dioxide (Aerosil)® 1.5 mg Cellulose, microcryst. (Avicel)® 70 mg

Modified cellulose gum (Ac-Di-Sol)® 7.5 mg Magnesium stearate Ad. Coating:

HPMC approx. 9 mg *Mywacett 9-40 T approx. 0.9 mg

*Acylated monoglyceride used as plasticizer for film coating.

A typical capsule for oral administration contains compounds of the invention (250 mg), lactose (75 mg) and magnesium stearate (15 mg). The mixture is passed through a 60 mesh sieve and packed into a No. 1 gelatin capsule. A typical injectable preparation is produced by aseptically placing 250 mg of compounds of the invention into a vial, aseptically freeze-drying and sealing. For use, the contents of the vial are mixed with 2 mL of sterile physiological saline, to produce an injectable preparation. The compounds of the invention may be administered to a mammal, especially a human in need of such treatment, prevention, elimination, alleviation or amelioration of the various diseases as mentioned above, e.g., a myeloproliferative condition, or cancer. Such mammals include also animals, both domestic animals, e.g. household pets, farm animals, and non-domestic animals such as wildlife.

The compounds of the invention are effective over a wide dosage range. For example, in the treatment of adult humans, dosages from about 0.05 to about 5000 mg, preferably from about 1 to about 2000 mg. A typical dosage is about 10 mg to about 1000 mg per day. In choosing a regimen for patients it may frequently be necessary to begin with a higher dosage and when the condition is under control to reduce the dosage. The exact dosage will depend upon the activity of the compound, mode of administration, on the therapy desired, form in which administered, the subject to be treated and the body weight of the subject to be treated, and the preference and experience of the physician or veterinarian in charge. JAK2 kinase inhibitor activity of the compounds of the invention may be determined by use of an in vitro assay system which measures the potentiation of inhibition of the JAK2 kinase. Inhibition constants (i.e., K, or IC₅₀ values as known in the art) for the JAK2 kinase inhibitors of the invention may be determined by the method described in the Examples.

Generally, the compounds of the invention are dispensed in unit dosage form comprising from about 10 mg to about 1000 mg of active ingredient together with a pharmaceutically acceptable carrier per unit dosage. Usually, dosage forms suitable for oral, nasal, pulmonal or transdermal administration comprise from about 125 μg to about 1250 mg, preferably from about 250 μg to about 500 mg, and more preferably from about 2.5 mg to about 250 mg, of the compounds admixed with a pharmaceutically acceptable carrier or diluent. The invention also encompasses prodrugs of a compound of the invention which on administration undergo chemical conversion by metabolic or other physiological processes before becoming active pharmacological substances. Conversion by metabolic or other physiological processes includes without limitation enzymatic (e.g, specific enzymatically catalyzed) and non- enzymatic (e.g., general or specific acid or base induced) chemical transformation of the prodrug into the active pharmacological substance. In general, such prodrugs will be functional derivatives of a compound of the invention which are readily convertible in vivo into a compound of the invention. Conventional procedures for the selection and preparation of suitable prodrug derivatives are described, for example, in Design of Prodrugs, ed. H. Bundgaard, Elsevier, 1985.

In another aspect, there are provided methods of making a composition of a compound described herein comprising formulating a compound of the invention with a pharmaceutically acceptable carrier or diluent. In some embodiments, the pharmaceutically acceptable carrier or diluent is suitable for oral administration. In some such embodiments, the methods may further comprise the step of formulating the composition into a tablet or capsule. In other embodiments, the pharmaceutically acceptable carrier or diluent is suitable for parenteral administration. In some such embodiments, the methods further comprise the step of lyophilizing the composition to form a lyophilized preparation.

B. Combinations. The compounds of the invention may be used in combination with i) one or more other JAK2 kinase inhibitors and/or ii) one or more other types of therapeutic agents, which may be administered orally in the same dosage form, in a separate oral dosage form (e.g., sequentially or non-sequentially) or by injection together or separately (e.g., sequentially or non-sequentially). Accordingly, in another aspect the invention provides combinations, comprising: a) a compound of the invention as described herein; and b) one or more compounds comprising: i) other compounds of the present invention ii) anticancer agents iii) anti-proliferative agents iv) immune modulators.

Combinations of the invention include mixtures of compounds from (a) and (b) in a single formulation and compounds from (a) and (b) as separate formulations. Some combinations of the invention may be packaged as separate formulations in a kit. In some embodiments, two or more compounds from (b) are formulated together while a compound of the invention is formulated separately. Combinations of the invention can further comprise a pharmaceutically acceptable carrier. In some embodiments, the compound of the invention is 90 wt % or more of a single diastereomer or single enantiomer. Alternatively, the compound of the invention can be 91, 92, 93, 94, 95, 96, 97, 98, or 99 wt % or more of a single diastereomer or single enantiomer.

The dosages and formulations for the other antiviral agent to be employed, where applicable, will be as set out in the latest edition of the Physicians' Desk Reference.

In carrying out the methods of the invention, a composition may be employed containing the compounds of the invention, with or without another antiviral agent and/or other type therapeutic agent, in association with a pharmaceutical vehicle or diluent. The composition can be formulated employing conventional solid or liquid vehicles or diluents and pharmaceutical additives of a type appropriate to the mode of desired administration. The compounds can be administered to mammalian species including humans, monkeys, dogs, etc. by an oral route, for example, in the form of tablets, capsules, granules or powders, or they can be administered by a parenteral route in the form of injectable preparations. The dose for adult humans is preferably between 10 and 1,000 mg per day, which can be administered in a single dose or in the form of individual doses from 1-4 times per day.

Examples

Synthetic Procedures for pyrazolo[l,5-a]pyrimidines:

LDA, THF Hydrazine hydrate

Ethylformate

Ethyl propiolate

5-10

Compound 2:

To a solution of diisopropyl amine (2.52 ml, 18 mmol) in tetrahydrofuran

(10 ml) at -78°C was added dropwise n-butyllithium (1.6M in hexanes, 11.2 ml,

18 mmol). The reaction was stirred at -78°C for 30 min. 1 (2.73 g, 18 mmol) was added and the reaction stirred for 10 min. The reaction mixture was added to a solution of ethyl formate (1.4 g, 19 mmol) in THF (5 ml) at -78°C. The reaction

was stirred at -78°C for 30min, then allowed to warm to r.t. and stirred for 16 h. The reaction was diluted with aqueous HCl (30 ml, IM) until the pH was approximately 3. The product was extracted with ethyl acetate(l 00ml*3). The combined organic fractions were washed with brine then dried over sodium sulphate and concentrated in vacuo. The product was purified on silica gel eluting with 1% MeOH in CH₂Cl₂, to give 2 as a white solid (2.9 g, 90%). LC- MS (ESI, positive): 180 [M+H]⁺.

Compound 3: To a solution of 2 (1.53 g, 8.52 mmol) in ethanol (25 ml) was added hydrazine hydrate (0.6 ml, 11.08 mmol) and acetic acid (0.83 ml, 14.48 mmol). The reaction was heated under reflux for 16 h. The solution was diluted with aqueous sodium bicarbonate (30 ml) and the product extracted with dichloromethane (60 mP2). The combined organic phase were washed with brine then dried over sodium sulphate and concentrated in vacuo to yield 3 as the white solid (1.6 g 97%). LC-MS (ESI, positive): 194 [M+H]⁺.

Compound 4:

To a stirred solution of 3 (3 g, 15.5 mmol) in 40ml of dioxane was added ethyl propiolate (1.65 g, 16.8 mmol) dropwise, and the resultant light yellow solution was warmed to reflux. After 1 Oh, the red solution was cooled to r.t. and 50 ml of toluene was added with stirred. The resultant precipitate was collected and purified on silica gel eluting with 3% MeOH in CH₂Cl₂ to afford 4 as a white solid (550mg, 14.4%). LC-MS (ESI, positive): 246 [M+H]⁺.

Compound 5:

To a stirred solution of 4 (50 mg 0.203 mmol) in 30 ml anhydrous CH₃CN in water-ice bath was added (CF₃SO₂)₂O (86 mg, 0.305 mmol) and TEA (41 mg, 0.406 mmol) dropwise respectively. After the total conversion of the start material detected by TLC, 2-(aminoethyl)pyridine(50mg, 0.406 mmol) and anhydrous K₂CO₃ (84 mg, 0.609 mmol) were added and the mixture was stirred at room temperature overnight. The solution was concentrated in vacuo and purified on silica gel eluting with 5% MeOH in CH₂Cl₂, to give the 5 (16mg,

22.6%). LC-MS (ESI, positive): 350 [M+H]⁺.; HNMR(CDCl₃): δ 1.684-1.661(d, 3H), 5.400-5.355(m, IH), 6.114-6.088(d, IH), 6.672-6.655(m, IH), 7.335- 7.130(m, 3H), 7.530-7.504(d, IH), 7.767-7.703(m, 2H), 8.201-8.130(m, 3H), 8.591-8.567(m, IH). Compound 6:

To a stirred solution of 4 (150 mg 0.609 mmol) in 50 ml anhydrous CH₃CN in water-ice bath was added (CF₃SO₂)₂O (258 mg, 0.914 mmol) and TEA (123 mg, 1.218 mmol) dropwise respectively. After the total conversion of the start material detected by TLC, S-2-(aminoethyl)fluorobenzene(169mg, 1.218 mmol) and anhydrous K₂CO₃ (252 mg, 1.827 mmol) were added and the mixture was stirred at room temperature overnight. The solution was concentrated in vacuo and purified on silica gel eluting with 3% MeOH in CH₂Cl₂, to give 6 (115mg, 51.6%). LC-MS (ESI, positive): 367 [M+H]⁺.; HNMR(CDCl₃): δ 1.540-1.517(d, 3H), 5.153(br. s, IH), 5.627-5.607(d, IH),

5.993-5.968(d, IH), 7.025-6.968(m, 2H), 7.131-7.105(m, IH), 7.271-7.219(m, IH), 7.408-7.364(m, 2H), 7.669-7.644(d, IH), 8.124-8.057(m, 3H).

Compound 7: To a stirred solution of 4 (150 mg 0.609 mmol) in 50 ml anhydrous

CH₃CN in water-ice bath was added (CF₃SO₂)₂O (258 mg, 0.914 mmol) and TEA (123 mg, 1.218 mmol) dropwise respectively. After the total conversion of the start material detected by TLC, R-2-(aminoethyl)fluorobenzene(169mg, 1.218 mmol) and anhydrous K₂CO₃ (252 mg, 1.827 mmol) were added and the mixture was stirred at room temperature overnight. The solution was concentrated in vacuo and purified on silica gel eluting with 3% MeOH in CH₂Cl₂, to give 7 (108mg, 48.5%). LC-MS (ESI, positive): 367 [M+H]⁺;

HNMR(CDCl₃): δ 1.568-1.545(d, 3H), 5.173(br. s, IH), 5.527-5.507(d, IH),

6.018-5.993(d, IH), 7.044-6.986(m, 2H), 7.143-7.113(m, IH), 7.284-7.232(m, IH), 7.429-7.383(m, 2H), 7.677-7.65 l(d, IH), 8.136-8.098(m, 3H).

Synthetic Procedures for 1,2,4-triazolopyridines:

11

12 13

14 15

Compound 11

2,6-dibromopyridine (4.0 g, 16.9 mmol) was dissolved in ammonia (30 ml) and the mixture was stirred in high-pressure autoclave at 120 ⁰C for 4 hours, the reaction mixture was allowed to cool, diluted with water, which was then extracted with CH₂Cl₂ (2x80 ml), then the organic phase was evaporated and the further material was chromatographed in ethylacetate : hexanes (1 :5) to afford 900mg of compound 11 as white solid. LC-MS (ESI, positive): 175 [M+H]⁺.

Compound 12

6-bromopyridin-2-amine (11, 900.0 mg, 5.2 mmol) was dissolved in 1,4- dioxane (20 ml) and the flask flushed with nitrogen. Ethoxycarbonyl isothiocyanate (682.3 mg, 5.2 mmol) was then added drop wise and the reaction mixture stirred at room temperature for 15 hours. The reaction solvent was removed under reduced pressure to give a solid, which was purified by silica gel (ethylacetate : hexanes 30%) to give 1.4 g of compound 12 as white solid. LC- MS (ESI, positive): 306 [M+H]⁺.

Compound 13

To a stirred suspension of hydroxylamine hydrochloride (959.1 g, 13.8 mmol) and DIPEA (1.2 g, 9.2 mmol) in methanol/ethanol (1 : 1 , 50 ml) was added compound 12 (1.4 g, 4.6 mmol) as solid. The mixture was stirred at room temperature for 2 hours, followed by 3hours at 60⁰C. The reaction mixture was

allowed to cool, diluted with CH₂Cl₂, which was then washed with water (2^χ80 ml), the organic phase was evaporated and the residue was chromatographed in MeOH/ CH₂Cl₂ (8% -10%) to afford 750 mg of compound 13 as a white solid. LC-MS (ESI, positive): 215 [M+H]⁺.

Compound 14

Compound 13 (750 mg, 3.5 mmol) was dissolved in toluene/ethanol (1 :1, 20 ml), to which was added 2M aqueous Na₂CO₃ (3.5 ml) and 3- chlorophenylboronicacid (1.1 g, 7.0 mmol). Pd (PPh₃) ₄ (202 mg) was added last, the mixture was flushed with nitrogen and refluxed for 5 hours in a sealed tube. The reaction mixture was allowed to cool.The reaction solvent was removed and the residue was purified on silica gel (ethylacetate : hexanes 50%) to give 514mg of compound 14 as white solid. LC-MS (ESI, positive): 247 [M+H]⁺.

Compound 15

Compound 14 (30mg, 0.12mmol) and pyridine (0.01ml, 0.12mmol 1) were dissolved in CH₂Cl₂ (8ml) and the flask flushed with nitrogen. Cyclobutanecarbonyl chloride (14.5mg, 0.2mmol) was then added dropwise and the reaction mixture stirred at 0 ⁰C for 8 hours. The reaction solvent was removed under reduced pressure to give a solid, which was purified by silica gel (ethylacetate : hexanes 50%) to yield 27mg of compound 15 as white solid. LC- MS (ESI, positive): 327 [M+H]⁺. Compounds 16-18 can be prepared by analogous methods to compound 15 using appropriately substituted reagents.

Table 1

/w Vitro TR-FRET JAK2 Kinase Assay

Compound activity to inhibit JAK2 mediated phosphorylation is determined by a time-resolved fluorescent resonance energy transfer (TR-FRET) assay using the LANCE® Eu-W 1024 labeled anti-phosphotyrosine (PT66) antibody as the donor and SureLight™ Allophycocyanin-Streptavidin (APC-SA) as the acceptor. The recombinant human JAK2 catalytic domain (826-1132 amino acids of accession # NP 004963.1) is expressed as an N-terminal His- tagged protein using a baculovirus expression system. A biotinylated Lyn peptide is used as the substrate. The JAK2 enzyme, substrate, and buffer were purchased from Carna Biosciences (Kobe, Japan). The PT66 antibody and the APC-SA fluorescent protein were purchased from Perkin Elmer (Waltham, MA).

Compounds were Vi logi₀ serially diluted in assay buffer containing 4% DMSO and are added to a 384- well black polystyrene assay plate with square wells. A substrate solution containing adenosine triphosphate (ATP) and the JAK2 enzyme were added to the assay plate to initiate the reaction. The final ATP concentration in the reaction mixture was 100 uM. The plate was shaken briefly, spun for 30-60 seconds at 1,000 rpm, and allowed to incubate at room temperature for 1 hour. A detection/stop solution was added to the plate containing 0.53 nM PT66, 33.3 nM APC-SA, 20 mM EDTA, 15 mM Tris-HCL (pH7.5), and 0.01% Tween20. The plate was incubated at room temperature, in the dark, for 30 minutes with gentle shaking. Phosphorylation of the substrate was detected by reading the plate in an Analyst HT 96.384 (Molecular Devices) microplate reader at an excitation wavelength of 360 nm and an emission of 665 nm. Percent inhibition values were calculated and graphed in a sigmoid dose response curve with a variable slope, using the non-linear regression program Prism (GraphPad Software, Inc.) to determine the 50% and 90% inhibitory concentrations (IC₅₀/IC₉₀).

Selected compounds of the invention have been found to have activity in this assay.

Claims

ClaimsWhat is claimed is:

1. A compound of formula (I):

(I) and hydrates, solvates, stereoisomers or mixtures thereof, tautomers, prodrugs, salts, crystalline forms, and mixtures thereof, wherein:

A, B, D, and E is each independently a carbon atom or a nitrogen atom, wherein R² is absent when A is a nitrogen atom and R³ is absent when E is a nitrogen atom, provided that least two of A, B, D, and E are nitrogen atoms and that one but not both of D and B is a nitrogen atom, and provided that neither D nor B comprises a quaternarized nitrogen atom; each dashed line represents a bond that can be present or absent, provided that bonds are present or absent so as to provide a bicyclic aromatic ring system; R¹ comprises hydrogen, alkyl, cycloalkyl, cycloalkylalkyl, aryl, aralkyl, heterocyclyl, heterocyclylalkyl, heteroaryl, heteroarylalkyl, C(O)R⁷, C(O)OR⁷, SO ₂R⁷ or C(O)NR⁸R⁹, wherein any alkyl, aryl, aralkyl, heterocyclyl, heterocyclylalkyl, heteroaryl or heteroarylalkyl is substituted with 0-3 J groups, or,

R¹ comprises -CH(R¹ ')(R¹²), wherein R^{1 1} is independently hydrogen, (C_rCi₂)-alkyl, (C₂-C ₁₂)-alkenyl, (C₂-C ₁₂)-alkynyl, (C₃-C i₀)-cycloalkyl or (C₃- Cio)-cycloalkenyl, [(C₃-C i₀)cycloalkyl or (C₃-Ci₀)-cycloalkenyl]-(C]-Ci₂)-alkyl, (C₆-C,o)-aryl, (C₆-Cio)-aiyl-(C,-C₁₂)-alkyl, (C₃-C ₁₀)-heterocyclyl, (C₃-C₁₀)- heterocyclyl-(C]-C₁₂)-alkyl, (C₅-C io)-heteroaryl, or (C₅-C]₀)-heteroaryl-(Ci-Ci₂)- alkyl, wherein R¹ ' other than hydrogen is substituted with 0-3 substituents selected independently from J, and R¹² is independently hydrogen or (Ci-C₆)- alkyl substituted with 0-3 substituents selected independently from J; R², R³ and R⁴ are independently hydrogen, alkyl, cycloalkyl, cycloalkylalkyl, aryl, aralkyl, heterocyclyl, heterocyclylalkyl, heteroaryl, heteroarylalkyl, C(O)R⁷, C(O)OR⁷, SO ₂R⁷ or C(O)NR⁸R⁹, wherein any alkyl, aryl, aralkyl, heterocyclyl, heterocyclylalkyl, heteroaryl or heteroarylalkyl is substituted with 0-3 J groups;

R⁵ and R⁶ are independently halogen, CN, CF₃, OCF₃, alkyl, cycloalkyl, cycloalkylalkyl, aryl, aralkyl, heterocyclyl, heterocyclylalkyl, heteroaryl or heteroarylalkyl, wherein any alkyl, cycloalkyl, cycloalkylalkyl, aryl, aralkyl, heterocyclyl, heterocyclylalkyl, heteroaryl or heteroarylalkyl is substituted with 0-3 J;

R⁷ is hydrogen, alkyl, cycloalkyl, cycloalkylalkyl, aryl, aralkyl, heterocyclyl, heterocyclylalkyl, heteroaryl or heteroarylalkyl, wherein any alkyl, cycloalkyl, cycloalkylalkyl, aryl, aralkyl, heterocyclyl, heterocyclylalkyl, heteroaryl or heteroarylalkyl is substituted with 0-3 J;

X and Y are each independently present or not present, and if present are independently C(R⁸)( R⁹), N(R¹⁰), or O, wherein

R and R are independently at each occurrence hydrogen, alkyl, cycloalkyl, cycloalkylalkyl, aryl, aralkyl, heterocyclyl, heterocyclylalkyl, heteroaryl or heteroarylalkyl, wherein any alkyl, cycloalkyl, cycloalkylalkyl, aryl, aralkyl, heterocyclyl, heterocyclylalkyl, heteroaryl or heteroarylalkyl is substituted with 0-3 J, or R and R , together with a carbon atom or a nitrogen atom to which they are bound, form a mono- or bicyclic ring system substituted with 0-3 J groups;

R is hydrogen, alkyl, aryl, aralkyl, alkoxy, cycloalkyl, cycloalkylidenyl, heterocyclyl, heterocyclylalkyl, heterocyclylalkylidenyl, or heteroaryl, wherein any R¹⁰ except hydrogen is substituted with 0-3 J groups; and,

J is halogen, alkyl, OR', CN, CF₃, OCF₃, R¹, O, S, C(O), S(O), methylenedioxy, ethylenedioxy, N(R')₂, SR', SOR', SO₂R', SO₂N(R')₂, SO₃R', C(O)R', C(O)C(O)R', C(O)CH₂C(O)R', C(S)R', C(O)OR', OC(O)R', C(0)N(R')₂, OC(O)N(R')₂, C(S)N(R')₂, (CH₂)_O-2NHC(O)R', N(R')N(R')C(0)R', N(R')N(R')C(0)0R', N(R')N(R')CON(R')₂, N(R')SO₂R', N(R')SO₂N(R')₂, N(R')C(0)0R', N(R')C(O)R', N(R')C(S)R^T, N(R')C(0)N(R')₂, N(R')C(S)N(R')₂, N(COR')COR', N(OR')R\ C(=NH)N(R')₂, C(O)N(OR')R', or C(=NOR')R' wherein; each R' is independently at each occurrence hydrogen, (Ci-Ci₂)-alkyl, (C₂-C_]2)alkenyl, (C₂-Ci₂)alkynyl, (C₃-C, ₀)-cycloalkyl, (C₃-C i₀)-cycloalkenyl, [(C₃-C,o)cycloalkyl, (C₃-C₁₀)-cycloalkenyl]-(C,-Ci₂)-alkyl, (C₆-C, ₀)-aryl, (C₆- C , o)-aryl-(C , -C , ₂)-alkyl, (C₃-C , ₀)-heterocyclyl, (C₃-C , ₀)-heterocyclyl-(C , -C , ₂)- alkyl, (C₅-C io)-heteroaryl, or (C₅-C, o)-heteroaryl-(C,-C,₂)-alkyl, wherein R' is substituted with 0-3 substituents selected independently from J, or, when two R' are bound to a nitrogen atom or to two adjacent nitrogen atoms, the two R' groups together with the nitrogen atom or atoms to which they are bound can form a 3- to 8-membered monocyclic ring, or an 8- to 20- membered bi- or tricyclic heterocyclic ring system, wherein any ring can further contain 1-3 additional heteroatoms selected from the group consisting of N, NR', NH, O, S, SO and SO₂ and wherein each ring is substituted with 0-3 substituents selected independently from J; wherein, in the bi- and tricyclic ring system, each ring is linearly fused, bridged, or spirocyclic, wherein each ring is either aromatic or nonaromatic, wherein each ring can be fused to a (C₆-C ,o)aryl, (C₅-C ,o)heteroaryl, (C₃- Cio)cycloalkyl or (C₃-C ,o)heterocyclyl.

2. A compound of claim 1, comprising a compound of formula (II):

wherein

X is present or not and if present can be independently C(R )( R ) or

N(R¹⁰); Y is C(R⁸X R⁹), N(R¹⁰), or O; R¹¹ is independently hydrogen, (Ci-Ci₂)-alkyl, (C₂-C i₂)-alkenyl, (C₂- Ci₂)-alkynyl, (C₃-C io)-cycloalkyl or (C₃-C io)-cycloalkenyl, [(C₃-C i₀)cycloalkyl or (C₃-C,o)-cycloalkenyl]-(C,-Ci₂)-alkyl, (C₆-C, ₀)-aryl, (C₆-C ₁₀)-aryl-(C ,-Ci₂)- alkyl, (C₃-C io)-heterocyclyl, (C₃-Ci₀)-heterocyclyl-(Ci-C,₂)-alkyl, (C₅-C₁₀)- heteroaryl, or (C₅-Ci₀)-heteroaryl-(Ci-Ci₂)-alkyl, wherein R¹ ' other than hydrogen is substituted with 0-3 substituents selected independently from J, and R¹² is independently hydrogen or (Ci-C₆)-alkyl substituted with 0-3 substituents selected independently from J¹, wherein each J¹ is independently halogen, OR', CN, CF₃, or OCF₃.

3. The compound of claim 1 , wherein B and E are each a nitrogen atom, and A and D are each a carbon atom.

4. The compound of claim 1, wherein A and D are each a nitrogen atom, and B and E are each a carbon atom.

5. The compound of any one of claims 1-4 wherein at least one of X and Y comprises NR¹⁰.

6. The compound of claim 3 wherein Y is absent and R¹ is hydrogen.

7. The compound of claim 3 or claim 6 wherein X is NR¹⁰.

8. The compound of claim 6 wherein R⁴ is C(O)R⁷.

9. The compound of claim 6 wherein R ° is H.

10. The compound of claim 8 wherein R⁷ is alkyl or cycloalkyl.

11. The compound of claim 3 wherein R² comprises an aryl group.

12. The compound of claim 11 wherein R² comprises a m-chlorophenyl group or a 2-methoxy-5-isopropylphenyl group.

13. The compound of claim 4 wherein X is absent and R⁴ is hydrogen.

14. The compound of claim 4 wherein Y is NR 10.

15. The compound of claim 14 wherein R¹ comprises an aryl, aralkyl, heteroaryl, or heteroarylalkyl group.

16. The compound of claim 1 comprising:

16. The compound of any one of claims 1-15 wherein the compound inhibits the activity of JAK2 with an effective ICj₀ of less than about 10 μM.

17. The compound of claim 16 wherein the JAK2 is a mutant JAK2.

18. The compound of claim 17 wherein the mutant JAK2 is JAK2V617F.

19. A method of treating a malcondition in a patient wherein inhibition of JAK2 is medically indicated, comprising providing a compound of any one of claims 1-15 to the patient in a dose, at a frequency, and for a duration sufficient to provide a beneficial effect to the patient.

20. The method of claim 19 wherein the JAK2 is a mutant JAK2.

21. The method of claim 20 wherein the mutant JAK2 is JAK2V617F.

22. The method of claim 19 wherein the malcondition comprises cancer or a myeloproliferative disease.

23. The method of claim 19 wherein the malcondition comprises an autoimmune disorder.

24. The method of claim 19 wherein modulation of cytokine signaling by inhibition of JAK2 is medically indicated.

25. A pharmaceutical composition comprising a compound of any one of claims 1-18 and a medically acceptable excipient.

26. A pharmaceutical combination comprising a compound of any one of claim 1-18 and a second medicament.

27. The combination wherein the second medicament comprises a second compound of formula (I), an anticancer agent, an anti-proliferative agent, or an immune modulator, or any combination thereof.

28. A pharmaceutical composition comprising the combination of claim 26 and a medically acceptable excipient.

29. A method of treating a malcondition in a patient wherein inhibition of JAK2 is medically indicated, comprising providing the combination of claim 26 to the patient in a dose, at a frequency, and for a duration for each medicament sufficient to provide a beneficial effect to the patient.

30. A method of inhibiting JAK2, comprising contacting a compound of any one of claims 1-18 with JAK2 at a concentration effective to bring about at least partial loss of activity of the JAK2.

31. A method of preparing a compound of claim 4, comprising contacting a compound of formula:

wherein Ar comprises an aryl group substituted with 0-3 J, with a compound of the formula R⁵-C≡C-C(O)OR" wherein R" comprises (C_rC₆)alkyl, to provide a compound of formula (III):

(HI), then, contacting the compound of formula (III) with a compound of formula (CF₃SO₂)₂O in the presence of an amine base to provide a compound of formula (IV):

(IV), then, contacting the compound of formula (IV) with a compound of formula R¹- YH, wherein -YH is -NH₂, to provide a compound of formula (I) of claim 4 wherein R⁶ is H and Y is NH.

32. A method of preparing a compound of claim 3, comprising contacting a compound of formula:

with hydroxylamine in the presence of an amine to provide a compound of formula (V):

(V) then, contacting the compound of formula (V) with an aryl boronic acid in the presence of a zero-valent palladium complex to provide a compound of formula:

wherein Ar comprises an aryl group substituted with 0-3 J.