The invention relates in part to molecules having certain biological activities that include, but are not limited to, inhibiting cell proliferation, and modulating certain protein kinase activities. Molecules of the invention modulate, e.g., Protein Kinase CK2 (called CK2 herein) and are useful to treat conditions associated directly or indirectly with CK2 activities, e.g., cancers, inflammatory conditions, infectious disorders, pain, immunological disorders, and neurodegenerative disorders (such as Alzheimer's disease and Parkinson's disease). The invention also relates in part to the use of such compounds, and pharmaceutical compositions containing these compounds.

WO2004/076458 describes pyrazolo[1,5-a]pyrimidines having protein kinase inhibitory activity. These compounds are useful in treating disorders where a kinase is implicated, such as inflammatory disease and cancer.

WO2004/028535 describes a series of thiazolidines that are modulators of the Pin1 protein and Pin1-related proteins.

Protein kinase CK2 (formerly called Casein kinase II, referred to herein as "CK2") is a ubiquitous and highly conserved protein serine/threonine kinase. The holoenzyme is typically found in tetrameric complexes consisting of two catalytic (alpha and/or alpha') subunits and two regulatory (beta) subunits. CK2 has a number of physiological targets and participates in a complex series of cellular functions including the maintenance of cell viability. The level of CK2 in normal cells is tightly regulated, and it has long been considered to play a role in cell growth and proliferation. Inhibitors of CK2 that are useful for treating certain types of cancers are described in PCT/US2007/077464, PCT/US2008/074820, PCT/US2009/35609.

The prevalence and importance of CK2, as well as an evolutionary analysis of its sequence, suggest it is an ancient enzyme on the evolutionary scale; its longevity may explain why it has become important in so many biochemical processes, and why CK2 from hosts have even been co-opted by infectious pathogens (e.g., viruses, protozoa) as an integral part of their survival and life cycle biochemical systems. These same characteristics explain why inhibitors of CK2 are believed to be useful in a variety of medical treatments as discussed herein. Because CK2 is central to many biological processes, as summarized by Guerra & Issinger, Curr. Med. Chem., 2008, 15:1870-1886, inhibitors of CK2, including the compounds described herein, should be useful in the treatment of a variety of diseases and disorders.

Cancerous cells show an elevation of CK2, and recent evidence suggests that CK2 exerts potent suppression of apoptosis in cells by protecting regulatory proteins from caspase-mediated degradation. The anti-apoptotic function of CK2 may contribute to its ability to participate in transformation and tumorigenesis. In particular, CK2 has been shown to be associated with acute and chronic myelogenous leukemia, lymphoma and multiple myeloma. In addition, enhanced CK2 activity has been observed in solid tumors of the colon, rectum and breast, squamous cell carcinomas of the lung and of the head and neck (SCCHN), adenocarcinomas of the lung, colon, rectum, kidney, breast, and prostate. Inhibition of CK2 by a small molecule is reported to induce apoptosis of pancreatic cancer cells, and hepatocellular carcinoma cells (HegG2, Hep3, HeLa cancer cell lines); and CK2 inhibitors dramatically sensitized RMS (Rhabdomyosarcoma) tumors toward apoptosis induced by TRAIL. Thus an inhibitor of CK2 alone, or in combination with TRAIL or a ligand for the TRAIL receptor, would be useful to treat RMS, the most common soft-tissue sarcoma in children. In addition, elevated CK2 has been found to be highly correlated with aggressiveness ofneoplasias, and treatment with a CK2 inhibitor of the invention should thus reduce tendency of benign lesions to advance into malignant ones, or for malignant ones to metastasize.

Unlike other kinases and signaling pathways, where mutations are often associated with structural changes that cause loss of regulatory control, increased CK2 activity level appears to be generally caused by upregulation or overexpression of the active protein rather than by changes that affect activation levels. Guerra and Issinger postulate this may be due to regulation by aggregation, since activity levels do not correlate well with mRNA levels. Excessive activity of CK2 has been shown in many cancers, including SCCHN tumors, lung tumors, breast tumors, and others. Id.

Elevated CK2 activity in colorectal carcinomas was shown to correlate with increased malignancy. Aberrant expression and activity of CK2 have been reported to promote increase nuclear levels of NF-kappaB in breast cancer cells. CK2 activity is markedly increased in patients with AML and CIVIL during blast crisis, indicating that an inhibitor of CK2 should be particularly effective in these conditions. Multiple myeloma cell survival has been shown to rely on high activity of CK2, and inhibitors of CK2 were cytotoxic to MM cells.

The literature provides clear evidence that inhibition of CK2 correlates with efficacy against tumor cells. For example, a CK2 inhibitor inhibited growth of murine p190 lymphoma cells. Its interaction with Bcr/Abl has been reported to play an important role in proliferation of Bcr/Abl expressing cells, indicating inhibitors of CK2 may be useful in treatment of Bcr/Abl-positive leukemias. Inhibitors of CK2 have been shown to inhibit progression of skin papillomas, prostate and breast cancer xenografts in mice, and to prolong survival of transgenic mice that express prostate-promoters. Id.

The role of CK2 in various non-cancer disease processes has been recently reviewed. See Guerra & Issinger, Curr. Med. Chem., 2008, 15:1870-1886. Increasing evidence indicates that CK2 is involved in critical diseases of the central nervous system, including, for example, Alzheimer's disease, Parkinson's disease, and rare neurodegenerative disorders such as Guam-Parkinson dementia, chromosome 18 deletion syndrome, progressive supranuclear palsy, Kuf's disease, or Pick's disease. It is suggested that selective CK2-mediated phosphorylation of tau proteins may be involved in progressive neurodegeneration of Alzheimer's disease. In addition, recent studies suggest that CK2 plays a role in memory impairment and brain ischemia, the latter effect apparently being mediated by CK2's regulatory effect on the PI3K survival pathways.

CK2 has also been shown to be involved in the modulation of inflammatory disorders, for example, acute or chronic inflammatory pain, glomerulonephritis, and autoimmune diseases, including, e.g., multiple sclerosis (MS), systemic lupus erythematosus, rheumatoid arthritis, and juvenile arthritis. It positively regulates the function of the serotonin 5-HT3 receptor channel, activates heme oxygenase type 2, and enhances the activity of neuronal nitric oxide synthase. A selective CK2 inhibitor was reported to strongly reduce pain response of mice when administered to spinal cord tissue prior to pain testing. It phosphorylates secretory type IIA phospholipase A2 from synovial fluid of RA patients, and modulates secretion of DEK (a nuclear DNA-binding protein), which is a proinflammatory molecule found in synovial fluid of patients with juvenile arthritis. Thus, inhibition of CK2 is expected to control progression of inflammatory pathologies such as those described here, and the inhibitors disclosed herein have been shown to effectively treat pain in animal models.

Protein kinase CK2 has also been shown to play a role in disorders of the vascular system, such as, e.g., atherosclerosis, laminar shear stress, and hypoxia. CK2 has also been shown to play a role in disorders of skeletal muscle and bone tissue, such as cardiomyocyte hypertrophy, impaired insulin signaling and bone tissue mineralization. In one study, inhibitors of CK2 were effective at slowing angiogenesis induced by growth factor in cultured cells. Moreover, in a retinopathy model, a CK2 inhibitor combined with octreotide (a somatostatin analog) reduced neovascular tufts; thus, the CK2 inhibitors described herein would be effective in combination with a somatoslatin analog to treat retinopathy.

CK2 has also been shown to phosphorylate GSK, troponin and myosin light chain; thus, CK2 is important in skeletal muscle and bone tissue physiology, and is linked to diseases affecting muscle tissue.

Evidence suggests that CK2 is also involved in the development and life cycle regulation of protozoal parasites, such as, for example, Theileria parva, Trypanosoma cruzi, Leishmania donovani, Herpetomonas muscarum muscarum, Plasmodium falciparum, Trypanosoma bucei, Toxoplasma gondii and Schistosoma mansoni. Numerous studies have confirmed the role of CK2 in regulation of cellular mobility of protozoan parasites, essential to invasion of host cells. Activation of CK2 or excessive activity of CK2 has been shown to occur in hosts infected with Leishmania donovani, Herpetomonas muscarum muscarum, Plasmodium falciparum, Trypanosoma brucei, Toxoplasma gondii and Schistosoma mansoni. Indeed, inhibition of CK2 has been shown to block infection by T. cruzi.

CK2 has also been shown to interact with and/or phosphorylate viral proteins associated with human immunodeficiency virus type 1 (HIV-1), human papilloma virus, and herpes simplex virus, in addition to other virus types (e.g. human cytomegalovirus, hepatitis C and B viruses, Borna disease virus, adenovirus, coxsackievirus, coronavirus, influenza, and varicella zoster virus). CK2 phosphorylates and activates HIV-1 reverse transcriptase and proteases in vitro and in vivo, and promotes pathogenicity of simian-human immunodeficiency virus (SHIV), a model for HIV. Inhibitors of CK2 are thus able to reduce pathogenic effects of a model of HIV infection. CK2 also phosphorylates numerous proteins in herpes simplex virus and numerous other viruses, and some evidence suggests viruses have adopted CK2 as a phosphorylating enzyme for their essential life cycle proteins. Inhibition of CK2 is thus expected to deter infection and progression of viral infections, which rely upon the host's CK2 for their own life cycles.

CK2 is unusual in the diversity of biological processes that it affects, and it differs from most kinases in other ways as well: it is constitutively active, it can use ATP or GTP, and it is elevated in most tumors and rapidly proliferating tissues. In addition, while many kinase inhibitors affect multiple kinases, increasing the likelihood of off-target effects or variability between individual subjects, CK2's unique structural features enable discovery of highly CK2-specific inhibitors. For all of these reasons, CK2 is a particularly interesting target for drug development, and the invention provides highly effective inhibitors of CK2 that are useful in treating a variety of different diseases and disorders mediated by or associated with excessive, aberrant or undesired levels of CK2 activity.

Compounds of Formula I have been found to be active on CK2 as well as on one or more Pim proteins. It has now been found that compounds of Formula (II) and (II') are typically more active on CK2, and also have less activity on Pim kinases. Without being bound by the theory, it is believed that their physiological activities derive from their activity on CK2.

The current invention provides novel compounds of Formula (II) and (II'), as well as Formulae IIa, IIa', II-Th and II-Th', and pharmaceutical compositions containing these compounds. The novel compounds of Formula II, which are related to the compounds of Formula I, show surprisingly greater activity on CK2 and reduced Pim activity, and thus are advantageously used to treat conditions sensitive to CK2 inhibition such as those described herein. Compounds of Formula II are therefore useful to treat conditions mediated by or associated with excessive activity of CK2, with reduced likelihood of off-target effects caused by inhibition of other kinases.

The present invention in part provides chemical compounds having certain biological activities that include, but are not limited to, inhibiting cell proliferation, inhibiting angiogenesis, and modulating protein kinase activities. These molecules modulate protein kinase CK2 (CK2) and/or PIM activity, and are typically more selective for CK2 activity over other kinases than similar compounds that lack the amine group shown in Formula (II) or (II'). These compounds affect biological functions that include but are not limited to, inhibiting gamma phosphate transfer from ATP to a protein or peptide substrate, inhibiting angiogenesis, inhibiting cell proliferation and inducing cell apoptosis, for example. Also provided are compositions comprising these molecules in combination with other materials, including other therapeutic agents, and methods for using such compositions.
Prior art compounds of the general formula (I) have been shown to inhibit Pim and CK2 (PCT/US2010/035657):

• wherein the bicyclic ring system containing Z¹-Z⁴ is aromatic;
• one of Z¹ and Z² is C, the other of Z¹ and Z² is N;
• Z³ and Z⁴ are independently CR⁵ or N,
  • where R⁵ can be H or R¹;
• R¹ is H, halo, CN, optionally substituted C1-C4 alkyl, optionally substituted C2-C4 alkenyl, optionally substituted C2-C4 alkynyl, optionally substituted C1-C4 alkoxy, or -NR⁷R⁸,
• where R⁷ and R⁸ are each independently selected from H, optionally substituted C1-C10 alkyl, optionally substituted aryl, optionally substituted arylalkyl, optionally substituted heteroaryl, and optionally substituted heteroarylalkyl,
• or R⁷ and R⁸ taken together with the N of -NR⁷R⁸ form an optionally substituted 5-8 membered ring that optionally contains an additional heteroatom selected from N, O and S as a ring member;
• R² is H, halo, CN, or an optionally substituted group selected from C1-C4 alkyl, C2-C4 alkenyl, and C2-C4 alkynyl;
• R³ and R⁴ are independently selected from H and optionally substituted C1-C10 alkyl;
• X is NR⁶, O, or S, where R⁶ is H or an optionally substituted group selected from C1-C4 alkyl, C2-C4 alkenyl, and C2-C4 alkynyl;
• Y is O or S;
• W is optionally substituted aryl, optionally substituted heteroaryl, or -NR⁹R¹⁰,-OR⁹, S(O)_nR⁹, optionally substituted carbon-linked heterocyclyl, optionally substituted C3-C8 cycloalkyl, or CR⁹R¹⁰R¹¹,
• wherein n is 0, 1 or 2, and
• R⁹ and R¹⁰ are each independently selected from H, optionally substituted C1-C10 alkyl, optionally substituted aryl, optionally substituted arylalkyl, optionally substituted heteroaryl, and optionally substituted heteroarylalkyl,
  • or R⁹ and R¹⁰ taken together with the N of -NR⁹R¹⁰ form an optionally substituted 5-8 membered ring that optionally contains an additional heteroatom selected from N, O and S as a ring member, and
• R¹¹ is selected from H, optionally substituted C1-C10 alkyl, optionally substituted aryl, optionally substituted arylalkyl, optionally substituted heteroaryl, and optionally substituted heteroarylalkyl.

The compounds of Formula I inhibit Pim and CK2, and often inhibit other kinases as well. For use as pharmaceuticals, it can be advantageous to select compounds that inhibit one primary target enzyme or receptor with minimal affect on other pathways or targets, because off-target biochemical effects can cause unpredictable side effects. It has now been found that novel compounds of Formula (II) and (II'), which are related to the compounds of Formula I, retain high levels of CK2 activity, and indeed are often more potent on CK2 than other compounds like Formula I, yet they are typically selective for CK2 over Pim kinases. In addition, their selectivity for CK2 over other kinases in a broad array of kinases is also improved over that of the compounds of Formula I generally. Therefore, compounds of Formula (II) or (II') represent a particularly useful class of compounds for the methods of treatment described herein, because they are selective for CK2 and inhibit fewer other kinases, resulting in a reduced risk of side-effects. The invention accordingly provides a compound which is a heterocycle of Formula (II), (II'), (IIa) or (IIa'): or a pharmaceutically acceptable salt and/or solvate thereof;
wherein, in formula (II), (II'), (IIa) and (IIa')

• Z³ represents N or CR⁵, or CH;
• Z⁴ represents CH, N or CR⁵
• each R⁵ is independently selected from halo, -CN, -R, -OR, -S(O)_nR, -COOR, -CONR₂, and -NR₂,
• wherein each R is independently selected from H and optionally substituted C1-C4 alkyl, or alternatively, the two R groups, taken together with the nitrogen atom to which they are attached, form an optionally substituted 5 or 6 membered heterocyclic ring that optionally contains one or more additional heteroatom selected from N, O and S as a ring member;
• each n is independently is 0,1 or 2; and
• each m is independently 1,2,3 or 4; in formulae (II) and (II'):
  • R², R³ and R⁴ are each independently selected from H and optionally substituted C1-C10 alkyl;
  • X represents NR², O, or S;
  • Y is O or S;
  • where R¹⁰ is selected from H, CN, optionally substituted C1-C4 alkyl, optionally substituted C2-C4 alkenyl, optionally substituted C2-C4 alkynyl, optionally substituted C1-C4 alkoxy, and -NR⁷R⁸,
  • ZisO;
  • L is -NR⁷-, a bond, -CR⁷=CR⁸-, -C≡C-, -O-, -S(O)_n-, -(CR⁷R⁸)_m-, -(CR⁷R⁸)_m-NR⁷-,-(CR⁷R⁸)_m-O-, or -(CR⁷R⁸)_m-S(O)_n-;
  • W is optionally substituted aryl, optionally substituted C1-C10 alkyl, optionally substituted C1-C10 heteroalkyl, optionally substituted heteroaryl, -NR⁷R⁸, -OR⁷, -S(O)_nR⁷,-CONR⁷R⁸, optionally substituted heterocyclyl, optionally substituted carbocyclyl, optionally substituted C2-C10 alkenyl, optionally substituted C2-C10 alkynyl, or -CR⁷R⁸R⁹;
  • where each R⁷ and R⁸ and R⁹ is independently selected from H, optionally substituted C1-C10 alkyl, optionally substituted heteroalkyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted carbocyclylalkyl, optionally substituted heterocyclylalkyl, optionally substituted aryl, optionally substituted arylalkyl, optionally substituted heteroaryl, and optionally substituted heteroarylalkyl;
  • or R⁸ and R⁹, taken together with the carbon atom to which they are attached, form =O (oxo) or =N-OR⁷ or =N-CN;
  • or R⁷ and R⁸, taken together on a single carbon atom or on adjacent connected carbon atoms of (CR⁷R⁸)_m whether alone or as part of another group, form a 3 to 8 membered carbocyclic ring or heterocyclic ring;
  • or R⁷ and R⁸, taken together with the nitrogen atom to which they are attached, form an optionally substituted 5 to 10 membered heterocyclic or heteroaryl ring that optionally contains one or more additional heteroatom selected from N, O and S as a ring member;
  • provided that no more than one of or R⁷ and R⁸ in -NR⁷R⁸ is selected from the group consisting of alkoxy, alkylamino, dialkylamino and heterocyclyl; and
  • R^1B is selected from optionally substituted carbocyclyl, H, optionally substituted C1-C10 alkyl, optionally substituted heteroalkyl, optionally substituted heterocyclyl, optionally substituted carbocyclylalkyl, optionally substituted heterocyclylalkyl, optionally substituted aryl, optionally substituted arylalkyl, optionally substituted heteroaryl, or optionally substituted heteroarylalkyl;
  • R^1A is selected from H, optionally substituted C1-C10 alkyl,
  • and in formulae (IIa) and (IIa')
  • R² is H, CH₃ or CF₃;
  • R⁴ is H, CH₃ or CF₃;
  • X, is O, S or NH;
  • Y is O or S;
  • R^1B is selected from H, optionally substituted C1-C10 alkyl, optionally substituted heteroalkyl, optionally substituted heterocyclyl, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted heterocyclylalkyl, optionally substituted arylalkyl, or an optionally substituted heteroaryl;
  • L is a bond, -NR⁷-, -O-, -S(O)_n-, (CR⁷R⁸)_m, or -(CR⁷R⁸)_m-NR⁷-;
  • W is selected from optionally substituted aryl, optionally substituted heteroaryl, and -NR⁷R⁸,
  • where each R⁷ and R⁸ is independently selected from H, optionally substituted C1-C6 alkoxy, optionally substituted C1-C6 alkylamino, optionally substituted C1-C6 dialkylamino, optionally substituted heterocyclyl, optionally substituted C1-C10 alkyl, optionally substituted C3-C8 cycloalkyl, optionally substituted C4-C10 cycloalkylalkyl, optionally substituted aryl, optionally substituted arylalkyl, optionally substituted heteroaryl, and optionally substituted heteroarylalkyl;
  • and R⁷ and R⁸, taken together on a single carbon atom or on adjacent connected carbon atoms of (CR⁷R⁸)_m whether alone or as part of another group, form a 3- to 8-membered ring that contains one or more heteroatoms as ring members;
  • or R⁷ and R⁸, taken together with the nitrogen atom to which they are attached, form an optionally substituted 5- to 10-membered heterocyclic or heteroaryl ring system that optionally contains an additional heteroatom selected from N, O and S as a ring member; and
  • provided that no more than one of or R⁷ and R⁸ in -NR⁷R⁸ is selected from the group consisting of alkoxy, alkylamino, dialkylamino and heterocyclyl.

A favored class of compounds of Formula II are those of Formula (IIa) or (IIa'): where R² is H, Me or CF₃; R⁴ is H, Me or CF₃; X is O, S or NH; Y is O or S; R^1B is as described for Formula II; L is a bond, -NR⁷-, -O-, or -S(O)_n-, (CR⁷R⁸)_m, or it can be -(CR⁷R⁸)_m-NR⁷-; m is 1-4 and n is 0-2; and W is selected from optionally substituted aryl, optionally substituted heteroaryl, and -NR⁷R⁸, where R⁷ and R⁸ are as defined for Formula II.

Particular embodiments of the compounds of the invention include thiophene-containing compounds of Formula (II-Th) and (II-Th'):

• where R^Th is selected from H, halo, optionally substituted C1-C6 alkyl, CN, S(O)_0-2R, -SO₂NR₂, COOR, CONR₂, and C(O)R,
• where each R is independently H, halo, CN, or an optionally substituted member selected from the group consisting of C1-C6 alkyl, C1-C6 alkoxy, C1-C6 alkylamino, di(C1-C6)alkylamino, C3-C8 cycloalkyl, C4-C10 cycloalkylalkyl, C5-C8 heterocyclyl, C6-C10 heterocyclylalkyl, aryl, arylalkyl, C5-C6 heteroalkyl, and C6-C10 heteroalkylalkyl;
• and two R on the same atom or adjacent atoms can form an optionally substituted heterocyclic ring that can contain an additional heteroatom selected from N, O and S; and other structural features are as defined for Formula IIa above.

The invention includes pharmaceutically acceptable salts of compounds of Formula II, II', IIa, IIa', II-Th, and II-Th' as well as the neutral compounds.

The invention also provides pharmaceutical compositions containing such compounds plus one or more pharmaceutically acceptable carriers or excipients, and the use of these compounds and compositions for the treatment of specified conditions as further described herein.

Intermediates of the following Formula (III) are useful for the preparation of compounds described above:

• where R^1A, R^1B, R², R⁴, Z³, Z⁴, L and W are as defined for Formula (II) above, or in certain embodiments, these are the same as the corresponding features defined for Formula (IIa) above;
• one of Z¹ and Z² represents N, and the other of Z¹ and Z² represents C;
• and the circles inside the two rings indicate that the rings are both aromatic.

The method of using these intermediates to produce compounds of formula (II) comprises reacting a compound of Formula (III) with a hydantoin or similar 5-membered heterocyclic compound of Formula (IV):

• where R³, X, Y and Z are as defined for Formula (II) or (II'),
• under conditions that promote condensation of the two compounds.

Typically, the reaction conditions will include a suitable solvent and a base, optionally a catalytic amount of base, but stoichiometric or larger amounts of base can be used. Suitable bases are those capable of deprotonating the compound of Formula (IV) to promote condensation with the compound of Formula (III), and secondary amines that are capable of reacting with aldehydes of Formula (III) to form an iminium species. Suitable bases include C1-C4 alkoxides, metal hydrides, tertiary amines such as triethylamine or diisopropyl ethylamine, DABCO and DBU; and suitable secondary amine bases include piperidine, morpholine, piperazine, N-methylpiperazine and pyrrolidine. Suitable solvents include polar aprotic solvents such as NMP, DMF, DMSO, DMA, and dioxane; as well as protic solvents such as C1-C10 alcohols and diols, e.g., ethanol, propanol, isopropanol, ethylene glycol, propylene glycol and methoxyethanol. Mixtures of such solvents can also be used, as can mixtures of one or more of these solvents with a less polar organic solvent to promote solubility of the reactants. Selection of suitable solvents and bases for these reactions are well within the level of skill of an ordinary practitioner.

In some embodiments of the compounds of Formula (III), -L-W represents a group of the formula -S(O)_1-2R, where R is an alkyl, cycloalkyl, aryl, heteroaryl or similar group, and the product is a compound of Formula (II) or (II') having the same -L-W group. Such compounds are conveniently used for the preparation of other compounds of formula (II) or (II'), because the moiety of formula -S(O)_1-2R is a good leaving group, and can readily be displaced by nucleophiles such as primary or secondary amines, to introduce other -L-W groups. Thus another method for synthesizing the compounds of the invention is to react a compound of Formula (V),

• wherein -L-W represents a group of the formula -S(O)_1-2R, where R is an optionally substituted group selected from C1-C6 alkyl, C3-C8 cycloalkyl, C4-C10 cycloalkylalkyl, C6-C10 aryl, C5-C6 heteroaryl, C7-C12 arylalkyl, and C6-C12 heteroarylalkyl;
• and other variables are as defined for formulas (III) and (IV) above;
• with a nucleophilic compound of formula
  
          W'-L'-H
  
  
  • wherein L' is selected from NR⁷, O and S; and
  • W' is optionally substituted aryl, optionally substituted heteroaryl optionally substituted heterocyclyl, optionally substituted C3-C8 cycloalkyl, optionally substituted C2-C10 alkenyl, optionally substituted C2-C10 alkynyl, or CR⁷R⁸R⁹,
  • where R⁷, R⁸ and R⁹ are as defined above for Formula II
    under suitable conditions as described herein to provide a compound of Formula (V'):

The invention also provides a pharmaceutical composition comprising a compound of Formula (II). (II'), (IIa) or (IIa') as described herein and at least one pharmaceutically acceptable carrier or excipient, or two or more pharmaceutically acceptable carriers and/or excipients. Pharmaceutical compositions comprising at least one of these compounds can be utilized in methods of treatment such as those described herein.

In common with the known compounds of Formula (I), the compounds of Formulae (II), (II'), (IIa) and (IIa') as described herein bind to and inhibit certain kinase proteins, which is believed to be the basis for their pharmaceutical activity. In certain embodiments, the protein is a CK2 protein, such as a CK2 protein comprising the amino acid sequence of SEQ ID NO:1, 2 or 3 or a substantially identical variant thereof, for example.

• SEQ ID NO:1 (NP 001886; casein kinase II alpha 1 subunit isoform a [Homo sapiens])
• SEQ ID NO:2 (NP 808227; casein kinase II alpha 1 subunit isoform a [Homo sapiens])
• SEQ ID NO:3 (NP 808228; casein kinase II alpha 1 subunit isoform b [Homo sapiens])

Substantially identical variants of these include proteins having at least 90% sequence homology with one of these, preferably at least 90% sequence identity; and having at least 50% of the level of in vitro kinase activity of the specified sequence under typical assay conditions.

Compounds of the invention can modulate the activity of CK2 protein, either in vitro or ex vivo. Suitable methods of such modulation comprise contacting a system comprising the protein with a compound described herein in an amount effective for modulating the activity of the protein. The invention accordingly further provides an in vitro method for modulating casein kinase 2 activity and/or Pim kinase activity in a cell, which comprises contracting the cell with a compound of the invention as defined above.

In certain embodiments the activity of the protein is inhibited, and sometimes the protein is a CK2 protein comprising the amino acid sequence of SEQ ID NO: 1, SEQ ID NO:2 or SEQ ID NO:3 or a substantially identical variant thereof, for example. In certain embodiments the CK2 is in a cell or tissue; in other embodiments, it can be in a cell-free system.

Compounds of the invention may also be used in methods for inhibiting cell proliferation, which comprise contacting cells with a compound described herein in an amount effective to inhibit proliferation of the cells. The cells sometimes are in a cell line, such as a cancer cell line (e.g., breast cancer, prostate cancer, pancreatic cancer, lung cancer, hemopoietic cancer, colorectal cancer, skin cancer, ovary cancer cell line), for example. In some embodiments, the cancer cell line is a breast cancer, prostate cancer or pancreatic cancer cell line. The cells sometimes are in a tissue, can be in a subject, at times are in a tumor, and sometimes are in a tumor in a subject. In one aspect the invention also provides a method for inhibiting cell proliferation, which comprises contacting cells in a cell line with a compound of the invention as defined above.

In certain embodiments, the method further comprises inducing cell apoptosis. Cells sometimes are from a subject having macular degeneration.

A compound described herein may be used in methods for treating a condition related to aberrant cell proliferation, which comprise administering such a compound to a subject in need thereof in an amount effective to treat the cell proliferative condition. In certain embodiments the cell proliferative condition is a tumor-associated cancer, e.g., a solid or circulating tumor. The cancer sometimes is cancer of the breast, prostate, pancreas, lung, colorectum, skin, or ovary. In some embodiments, the cell proliferative condition is a non-tumor cancer, such as a hematopoietic cancer, for example, including leukemias, e.g., multiple myeloma and lymphomas. The cell proliferative condition is macular degeneration in some embodiments.

Compounds of the invention as described above may be used in methods for treating cancer or an inflammatory disorder or other disorders described herein that are mediated by excessive activity of one or more of these kinases, in a subject in need of such treatment. Such methods comprise administering to the subject a therapeutically effective amount of a therapeutic agent useful for treating such disorder; and administering to the subject a molecule described herein, e.g., a compound inhibits CK2 in an amount that is effective to enhance a desired effect of the therapeutic agent.

Accordingly, the invention further provides a compound of formula (II), (II'), (IIa) or (IIa') as defined above, or a pharmaceutically acceptable salt or solvate thereof, for use in

1. (a) treating a condition or disease associated with casein kinase 2 activity and/or Pim kinase activity in a patient by administering to the patient a therapeutically effective amount of the compound, wherein
   the condition or disease is selected from cancer, a vascular disorder, a inflammation, a pathogenic infection, a immunological disorder, a neurodegenerative disorder, and a combination thereof, and wherein the cancer is preferably selected from cancer of the colorectum, breast, lung, liver, pancreas, lymph node, colon, prostate, brain, head and neck, skin, liver, kidney, blood and heart, and wherein the compound is optionally administered in combination with one or more additional therapeutic agent, preferably an anticancer agent; or
2. (b) inhibiting cell proliferation, which comprises contacting cells with the compound in an amount effective to inhibit proliferation of the cells, wherein the cells are in a tumor in a subject; or
3. (c) inhibiting angiogenesis in a subject, which comprises administering to the subject the compound in an amount effective to inhibit the angiogenesis.

The molecule that inhibits CK2 is a compound of Formula II, or Formula II' or (IIa) or (IIa'), or a pharmaceutically acceptable salt or solvate thereof. In certain embodiments, the desired effect of the therapeutic agent that is enhanced by the molecule that inhibits CK2 is an increase in apoptosis in at least one type of cell. In certain embodiments, the cell is a cancer cell and the compound is a compound of Formula (II) or (IIa) that is a potent inhibitor (IC-50 less than about 100 nM, for example) of CK2. Preferably, the compound has an IC-50 on Pim of less than about 30 nM, and is selective for CK2 over Pim kinases. In certain embodiments, the IC-50 for inhibition of CK2 is lower by at least a factor of ten than activity on Pim; in preferred embodiments, the compound has an IC-50 for CK2 that is lower than its IC-50 for at least one of Pim-1, Pim-2 and Pim-3 by about 100-fold or more.

In some embodiments, the therapeutic agent and the molecule that inhibits CK2 are administered at substantially the same time. The therapeutic agent and molecule that inhibits CK2 sometimes are used concurrently by the subject. The therapeutic agent and the molecule that inhibits CK2 can be combined into one pharmaceutical composition in certain embodiments; in other embodiments that are admistered as separate compositions.

Also provided are compositions of matter comprising a compound of the invention as described herein and an isolated protein. The protein sometimes is a CK2 protein, such as a CK2 protein comprising the amino acid sequence of SEQ ID NO:1, SEQ ID NO:2 or SEQ ID NO:3 or a substantially identical variant thereof, for example. In some embodiments, the protein is a Pim protein. Certain compositions comprise a compound described herein in combination with a cell. The cell may be from a cell line, such as a cancer cell line. In the latter embodiments, the cancer cell line is sometimes a breast cancer, prostate cancer, pancreatic cancer, lung cancer, hematopoietic cancer, colorectal cancer, skin cancer, of ovary cancer cell line.

The invention is further described in the description that follows. Embodiments falling outside the scope of claim are included for reference purposes.

• Figure 1 depicts a compound of Formula I as described herein, and shows its IC50 on CK2 (7 nM) and on PIM1 (351 nM), and also shows a plot of inhibition of a panel of 108 kinases to illustrate its selectivity for these kinases relative to other kinases.
• Figure 2 depicts a compound of Formula II as described herein, and shows that it is more potent on CK2 (3 nM), less potent on PIM1 (1310 nM), and generally more selective towards various kinases than is the compound in Figure 1.
• Figure 3 shows a synthesis scheme for preparing certain compounds of the invention containing a thiophene ring.
• Figure 4 illustrates the syntheses of certain pyrazolotriazines of the invention.
• Figure 5 illustrates synthesis methods for introducing various nucleophilic groups onto a pyrazolo-triazine ring system for preparing compounds of the invention.
• Figure 6 illustrates general synthesis routes for making certain pyrazolo-triazine compounds of the invention.
• Figure 7 shows a general synthetic method for making various imidazo-pyrazine ring systems and for making certain compounds of the invention.
• Figure 8 depicts a number of variations of the pyrazolo-triazine compounds within the scope of the invention.
• Figure 9 depicts methods to make certain imidazo-pyridazine compounds within the scope of the invention.
• Figure 10 illustrates a general method for modifying certain substituted compounds of the invention to introduce additional features.
• Figure 11 depicts more methods for modifying substituents on compounds of the invention.
• Figure 12 illustrates alternative synthesis routes for making certain compounds of the invention.
• Figure 13 depicts formation of an amide compound of the invention from a corresponding carboxylic acid compound.
• Figure 14 depicts a reductive amination method for introducing certain groups onto the compounds of the invention.

Compounds of the present invention exert biological activities that include, but are not limited to, inhibiting cell proliferation, reducing angiogenesis, preventing or reducing inflammatory responses and pain, and modulating certain immune responses. Such compounds modulate CK2 activity, as demonstrated by the data herein. Such compounds therefore can be utilized in multiple applications by a person of ordinary skill in the art. For example, compounds described herein can be used, for example, for (i) modulation of protein kinase activity (e.g., CK2 activity), (ii) modulation of cell proliferation, (iii) modulation of apoptosis, and/or (iv) treatments of cell proliferation related disorders (e.g., administration alone or coadministration with another molecule). In particular, the compounds of Formula (II) and (IIa) can be used to modulate CK2 activity, in vitro or in vivo, and to treat disorders associated with excessive or undesirable levels of CK2 activity, including cancers, certain inflammatory disorders, vascular disorders, certain skeletal and muscle disorders, and infections such as protozoal parasite infections and some viral infections.

The terms "a" and "an" do not denote a limitation of quantity, but rather denote the presence of at least one of the referenced item. The terms "a" and "an" are used interchangeable with "one or more" or "at least one". The term "or" or "and/or" is used as a function word to indicate that two words or expressions are to be taken together or individually. The terms "comprising", "having", "including", and "containing" are to be construed as open-ended terms (i.e., meaning "including, but not limited to"). The endpoints of all ranges directed to the same component or property are inclusive and independently combinable.

The terms "compound(s) of the invention", "these compounds", "such compound(s)", "the compound(s)", and "the present compound(s)" refer generally to compounds of the structural Formula (II), (II'), (IIa), (IIa'), (IIb), (IIb'), (IIc), (II-Th), and (II-Th'), and includes any specific compounds within these formulae whose structure is disclosed herein. Compounds may be identified either by their chemical structure and/or chemical name. When the chemical structure and chemical name conflict, the chemical structure is determinative of the identity of the compound. Furthermore, the present compounds can modulate, i.e., inhibit or enhance, the biological activity of a CK2 protein, a Pim protein or both, and thereby is also referred to herein as a "modulator(s)" or "CK2 and/or Pim modulator(s)". Compounds of Formulae (I), (II), (II'), (IIa), (IIa'), (IIb), (IIb'), (IIc); (II-Th), and
(II-Th'), including any specific compounds, i.e., species, described herein are exemplary "modulator".

The compounds described herein may contain one or more chiral centers and/or double bonds and therefore, may exist as stereoisomers, such as double-bond isomers (i.e., geometric isomers such as E and Z), enantiomers or diastereomers. The invention includes each of the isolated stereoisomeric forms (such as the enantiomerically pure isomers, the E and Z isomers, and etc.) as well as mixtures of stereoisomers in varying degrees of chiral purity or percetange of E and Z, including racemic mixtures, mixtures of diastereomers, and mixtures of E and Z isomers. Accordingly, the chemical structures depicted herein encompass all possible enantiomers and stereoisomers of the illustrated compounds including the stereoisomencally pure form (e.g., geometrically pure, enantiomerically pure or diastereomerically pure) and enantiomeric and stereoisomeric mixtures. Enantiomeric and stereoisomeric mixtures can be resolved into their component enantiomers or stereoisomers using separation techniques or chiral synthesis techniques well known to the skilled artisan. The invention includes each of the isolated stereoisomeric forms as well as mixtures of stereoisomers in varying degrees of chiral purity, including racemic mixtures. It also encompasses the various diastereomers. Other structures may appear to depict a specific isomer, but that is merely for convenience, and is not intended to limit the invention to the depicted olefin isomer. When the chemical name does not specify the isomeric form of the compound, it denotes any one of the possible isomeric forms or a mixtures of those isomeric forms of the compound.

The compounds may also exist in several tautomeric forms, and the depiction herein of one tautomer is for convenience only, and is also understood to encompass other tautomers of the form shown. Accordingly, the chemical structures depicted herein encompass all possible tautomeric forms of the illustrated compounds. The term "tautomer" as used herein refers to isomers that change into one another with great ease so that they can exist together in equilibrium. For example, ketone and enol are two tautomeric forms of one compound. In another example, a substituted 1.2,4-triazole derivative may exist in at least three tautomeric forms as shown below:

The descriptions of compounds of the present invention are limited by principles of chemical bonding known to those skilled in the art. Accordingly, where a group may be substituted by one or more of a number of substituents, such substitutions are selected so as to comply with principles of chemical bonding and to give compounds which are not inherently unstable and/or would be known to one of ordinary skill in the art as likely to be unstable under ambient conditions, such as aqueous, neutral, and several known physiological conditions. For example, a heterocycloalkyl or heteroaryl is attached to the remainder of the molecule via a ring heteroatom in compliance with principles of chemical bonding known to those skilled in the art thereby avoiding inherently unstable compounds.

The compounds of the invention often have ionizable groups so as to be capable of preparation as salts. In that case, wherever reference is made to the compound, it is understood in the art that a pharmaceutically acceptable salt may also be used. These salts may be acid addition salts involving inorganic or organic acids or the salts may, in the case of acidic forms of the compounds of the invention be prepared from inorganic or organic bases. Frequently, the compounds are prepared or used as pharmaceutically acceptable salts prepared as addition products of pharmaceutically acceptable acids or bases. Suitable pharmaceutically acceptable acids and bases are well-known in the art, such as hydrochloric, sulphuric, hydrobromic, acetic, lactic, citric, or tartaric acids for forming acid addition salts, and potassium hydroxide, sodium hydroxide, ammonium hydroxide, caffeine and various amines, for forming basic salts. Methods for preparation of the appropriate salts are well-established in the art. In some cases, the compounds may contain both an acidic and a basic functional group, in which case they may have two ionized groups and yet have no net charge. Standard methods for the preparation of pharmaceutically acceptable salts and their formulations are well known in the art, and are disclosed in various references, including for example, "Remington: The Science and Practice of Pharmacy", A. Gennaro, ed., 20th edition, Lippincott, Williams & Wilkins, Philadelphia, PA.

"Solvate", as used herein, means a compound formed by solvation (the combination of solvent molecules with molecules or ions of the solute), or an aggregate that consists of a solute ion or molecule, i.e., a compound of the invention, with one or more solvent molecules. When water is the solvent, the corresponding solvate is "hydrate". Examples of hydrate include, but are not limited to, hemihydrate, monohydrate, dihydrate, trihydrate, hexahydrate, etc. It should be understood by one of ordinary skill in the art that the pharmaceutically acceptable salt, and/or prodrug of the present compound may also exist in a solvate form. The solvate is typically formed via hydration which is either part of the preparation of the present compound or through natural absorption of moisture by the anhydrous compound of the present invention.

The term "ester" means any ester of a present compound in which any of the -COOH functions of the molecule is replaced by a -COOR function, in which the R moiety of the ester is any carbon-containing group which forms a stable ester moiety, including but not limited to alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkylalkyl, aryl, arylalkyl, heterocyclyl, heterocyclylalkyl and substituted derivatives thereof. The hydrolysable esters of the present compounds are the compounds whose carboxyls are present in the form of hydrolysable ester groups. That is, these esters are pharmaceutically acceptable and can be hydrolyzed to the corresponding carboxyl acid in vivo. These esters may be conventional ones, including lower alkanoyloxyalkyl esters, e.g. pivaloyloxymethyl and 1-pivaloyloxyethyl esters; lower alkoxycarbonylalkyl esters, e.g., methoxycarbonyloxymethyl, 1-ethoxycarbonyloxyethyl, and 1-isopropylcarbonyloxyethyl esters; lower alkoxymethyl esters, e.g., methoxymethyl esters, lactonyl esters, benzofuran keto esters, thiobenzofuran keto esters; lower alkanoylaminomethyl esters, e.g., acetylaminomethyl esters. Other esters can also be used, such as benzyl esters and cyano methyl esters. Other examples of these esters include: (2,2-dimethyl-1-oxypropyloxy)methyl esters; (1RS)-1-acetoxyethyl esters, 2-[(2-methylpropyloxy)carbonyl]-2-pentenyl esters, 1-[[(1-methylethoxy)carbonyl]- oxy]ethyl esters; isopropyloxycarbonyloxyethyl esters, (5-methyl-2-oxo-1,3- dioxole-4-yl) methyl esters, 1-[[(cyclohexyloxy)carbonyl]oxy]ethyl esters; 3,3-dimethyl-2-oxobutyl esters. It is obvious to those skilled in the art that hydrolysable esters of the compounds of the present invention can be formed at free carboxyls of said compounds by using conventional methods. Representative esters include pivaloyloxymethyl esters, isopropyloxycarbonyloxyethyl esters and (5-methyl-2-oxo-1,3-dioxole-4-yl)methyl esters.

"Protecting group" refers to a grouping of atoms that when attached to a reactive functional group in a molecule masks, reduces or prevents reactivity of the functional group. Examples of protecting groups can be found in Green et al., "Protective Groups in Organic Chemistry", (Wiley, 2nd ed. 1991) and Harrison et al., "Compendium of Synthetic Organic Methods", Vols. 1-8 (John Wiley and Sons, 1971-1996). Representative amino protecting groups include, but are not limited to, formyl, acetyl, trifluoroacetyl, benzyl, benzyloxycarbonyl ("CBZ"), tert-butoxycarbonyl ("Boc"), trimethylsilyl ("TMS"), 2-trimethylsilyl-ethanesulfonyl ("SES"), trityl and substituted trityl groups, allyloxycarbonyl, 9-fluorenylmethyloxycarbonyl and ("FMOC"), nitro-veratryloxycarbonyl ("NVOC"). Representative hydroxy protecting groups include, but are not limited to, those where the hydroxy group is either acylated or alkylated such as benzyl, and trityl ethers as well as alkyl ethers, tetrahydropyranyl ethers, trialkylsilyl ethers and allyl ethers.

As used herein, "pharmaceutically acceptable" means suitable for use in contact with the tissues of humans and animals without undue toxicity, irritation, allergic response, and the like, commensurate with a reasonable benefit/risk ratio, and effective for their intended use within the scope of sound medical judgment.

"Excipient" refers to a diluent, adjuvant, vehicle, or carrier with which a compound is administered.

An "effective amount" or "therapeutically effective amount" is the quantity of the present compound in which a beneficial outcome is achieved when the compound is administered to a patient or alternatively, the quantity of compound that possesses a desired activity in vivo or in vitro. In the case of proliferative disorders, a beneficial clinical outcome includes reduction in the extent or severity of the symptoms associated with the disease or disorder and/or an increase in the longevity and/or quality of life of the patient compared with the absence of the treatment. For example, for a subject with cancer, a "beneficial clinical outcome" includes a reduction in tumor mass, a reduction in the rate of tumor growth, a reduction in metastasis, a reduction in the severity of the symptoms associated with the cancer and/or an increase in the longevity of the subject compared with the absence of the treatment. The precise amount of compound administered to a subject will depend on the type and severity of the disease or condition and on the characteristics of the patient, such as general health, age, sex, body weight and tolerance to drugs. It will also depend on the degree, severity and type of proliferative disorder. The skilled artisan will be able to determine appropriate dosages depending on these and other factors.

As used herein, the terms "alkyl," "alkenyl" and "alkynyl" include straight-chain, branched-chain and cyclic monovalent hydrocarbyl radicals, and combinations of these, which contain only C and H when they are unsubstituted. Examples include methyl, ethyl, isobutyl, cyclohexyl, cyclopentylethyl, and 2-propenyl, 3-butynyl. The total number of carbon atoms in each such group is sometimes described herein, e.g., when the group can contain up to ten carbon atoms it can be represented as 1-10C or as C1-C10 or C1-10. When heteroatoms (N, O and S typically) are allowed to replace carbon atoms as in heteroalkyl groups, for example, the numbers describing the group, though still written as e.g. C1-C6, represent the sum of the number of carbon atoms in the group plus the number of such heteroatoms that are included as replacements for carbon atoms in the backbone of the ring or chain being described. Where a ring is included, it is understood that the group contains at least three carbon atoms as a 3-membered ring is the smallest size for a ring.

Typically, the alkyl, alkenyl and alkynyl substituents of the invention contain 1-10C (alkyl) or 2-10C (alkenyl or alkynyl), or 3-10C when a ring is included. Preferably they contain 1-8C (alkyl) or 2-8C (alkenyl or alkynyl) or 3-8C when a ring is included. Sometimes they contain 1-4C (alkyl) or 2-4C (alkenyl or alkynyl). A single group can include more than one type of multiple bond, or more than one multiple bond; such groups are included within the definition of the term "alkenyl" when they contain at least one carbon-carbon double bond, and are included within the term "alkynyl" when they contain at least one carbon-carbon triple bond; provided, however, that the presence of multiple bonds does not produce an aromatic ring.

Alkyl, alkenyl and alkynyl groups are often optionally substituted to the extent that such substitution makes sense chemically.

"Optionally substituted" as used herein indicates that the particular group or groups being described may have no non-hydrogen substituents, or the group or groups may have one or more non-hydrogen substituents. If not otherwise specified, the total number of such substituents that may be present is equal to the number of H atoms present on the unsubstituted form of the group being described. Where an optional substituent is attached via a double bond, such as a carbonyl oxygen (=O), the group takes up two available valences, so the total number of substituents that may be included is reduced according to the number of available valences.

"Substituted," when used to modify a specified group or radical, means that one or more hydrogen atoms of the specified group or radical are each, independently of one another, replaced with the same or different substituent(s).

Substituent groups useful for substituting saturated carbon atoms in the specified group or radical include, but are not limited to -R^a, halo, -O^-, =O, -OR^b, -SR^b, -S^-, =S, -NR^cR^c, =NR^b, =N-OR^b, trihalomethyl, -CF₃, -CN, -OCN, -SCN, -NO, -NO₂, =N₂, -N₃, -S(O)₂R^b, -S(O)₂NR^b, -S(O)₂O^-, -S(O)₂OR^b, -OS(O)₂R^b, -OS(O)₂O^-, -OS(O)₂OR^b, -P(O)(O^-)₂, -P(O)(OR^b)(O^-), -P(O)(OR^b)(OR^b), -C(O)R^b, -C(S)R^b, -C(NR^b)R^b, -C(O)O^-, -C(O)OR^b, -C(S)OR^b, -C(O)NR^cR^c, -C(NR^b)NR^cR^c, -OC(O)R^b, -OC(S)R^b, -OC(O)O^-, -OC(O)OR^b, -OC(S)OR^b, -NR^bC(O)R^b, -NR^bC(S)R^b, -NR^bC(O)O^-, -NR^bC(O)OR^b, -NR^bC(S)OR^b, -NR^bC(O)NR^cR^c, -NR^bC(NR^b)R^b and -NR^bC(NR^b)NR^cR^c, where R^a is selected from the group consisting of alkyl, cycloalkyl, heteroalkyl, cycloheteroalkyl, aryl, arylalkyl, heteroaryl and heteroarylalkyl; each R^b is independently hydrogen or R^a; and each R^c is independently R^b or alternatively, the two R^cs may be taken together with the nitrogen atom to which they are bonded form a 4-, 5-, 6- or 7-membered cycloheteroalkyl which may optionally include from 1 to 4 of the same or different additional heteroatoms selected from the group consisting of O, N and S. As specific examples, -NR^cR^c is meant to include -NH₂, -NH-alkyl, N-pyrrolidinyl and N-morpholinyl. As another specific example, a substituted alkyl is meant to include -alkylene-O-alkyl, -alkylene-heteroaryl, -alkylene-cycloheteroalkyl, -alkylene-C(O)OR^b, -alkylene-C(O)NR^bR^b, and -CH₂-CH₂-C(O)-CH₃. The one or more substituent groups, taken together with the atoms to which they are bonded, may form a cyclic ring including cycloalkyl and cycloheteroalkyl.

Similarly, substituent groups useful for substituting unsaturated carbon atoms in the specified group or radical include, but are not limited to, -R^a, halo, -O^-, -OR^b, -SR^b, -S^-, -NR^cR^c, trihalomethyl, -CF₃, -CN, -OCN, -SCN, -NO, -NO₂, -N₃, -S(O)₂R^b, -S(O)₂O^-, -S(O)₂OR^b, -OS(O)₂R^b, -OS(O)₂O^-, -OS(O)₂OR^b, -P(O)(O^-)₂, -P(O)(OR^b)(O^-), -P(O)(OR^b)(OR^b), -C(O)R^b, -C(S)R^b, -C(NR^b)R^b, -C(O)O^-, -C(O)OR^b, -C(S)OR^b, -C(O)NR^cR^c, -C(NR^b)NR^cR^c, -OC(O)R^b, -OC(S)R^b, -OC(O)O^-, -OC(O)OR^b, -OC(S)OR^b, -NR^bC(O)R^b, -NR^bC(S)R^b, -NR^bC(O)O^-, -NR^bC(O)OR^b, -NR^bC(S)OR^b, -NR^bC(O)NR^cR^c, -NR^bC(NR^b)R^b and -NR^bC(NR^b)NR^cR^c, where R^a, R^b and R^c are as previously defined.

Substituent groups useful for substituting nitrogen atoms in heteroalkyl and cycloheteroalkyl groups include, but are not limited to, -R^a, -O^-, -OR^b, -SR^b, -S^-, -NR^cR^c, trihalomethyl, -CF₃, -CN, -NO, -NO₂, -S(O)₂R^b, -S(O)₂O^-, -S(O)₂OR^b, -OS(O)₂R^b, -OS(O)₂O^-, -OS(O)₂OR^b, -P(O)(O^-)₂, -P(O)(OR^b)(O^-), -P(O)(OR^b)(OR^b), -C(O)R^b, -C(S)R^b, -C(NR^b)R^b, -C(O)OR^b, -C(S)OR^b, -C(O)NR^cR^c, -C(NR^b)NR^cR^c, -OC(O)R^b, -OC(S)R^b, -OC(O)OR^b, -OC(S)OR^b, -NR^bC(O)R^b, -NR^bC(S)R^b, -NR^bC(O)OR^b, -NR^bC(S)OR^b, -NR^bC(O)NR^cR^c, -NR^bC(NR^b)R^b and -NR^bC(NH^b)NR^cR^c, where R^a, R^b and R^c are as previously defined.

Alkyl, alkenyl and alkynyl groups can alternatively or in addition be substituted by C1-C8 acyl, C2-C8 heteroacyl, C6-C10 aryl, C3-C8 cycloalkyl, C3-C8 heterocyclyl, or C5-C10 heteroaryl, each of which can be substituted by one or more R, halo, =O, =N-CN, =N-OR, =NR, OR, NR₂, SR, SO₂R, SO₂NR₂, NRSO₂R, NRCONR₂, NRCSNR₂, NRC(=NR)NR₂, NRCOOR, NRCOR, CN, C≡CR, COOR, CONR₂, OOCR, COR, and NO₂, wherein each R is independently H, C1-C8 alkyl, C2-C8 heteroalkyl, C1-C8 acyl, C2-C8 heteroacyl, C2-C8 alkenyl, C2-C8 heteroalkenyl, C2-C8 alkynyl, C2-C8 heteroalkynyl, C3-C8 heterocyclyl, C4-C10 heterocyclylalkyl, C6-C10 aryl, or C5-C10 heteroaryl, and each R is optionally substituted with one or more (typically up to three) halo, =O, =N-CN, =N-OR', =NR', OR', NR'₂, SR', SO₂R', SO₂NR'₂, NR'SO₂R', NR'CONR'₂, NR'CSNR'₂, NR'C(=NR')NR'₂, NR'COOR', NR'COR', CN, C'≡CR', COOR', CONR'₂, OOCR', COR', and NO₂, wherein each R' is independently H, C1-C8 alkyl, C2-C8 heteroalkyl, C1-C8 acyl, C3-C8 heterocyclyl, C2-C8 heteroacyl, C6-C10 aryl or C5-C10 heteroaryl.

Where any of these substituents contains two R or R' groups on the same or adjacent atoms (e.g., -NR₂, or -NR-C(O)R), the two R or R' groups can optionally be taken together with the atom(s) in the substituent group to which they are attached to form a ring having 5-8 ring members, which can include another heteroatom as a ring member (N, O or S) and can be substituted with one or more halo, =O, =N-CN, =N-OR, =NR, OR, NR₂, SR, SO₂R, SO₂NR₂, NRSO₂R, NRCONR₂, NRCSNR₂, NRC(=NR)NR₂, NRCOOR, NRCOR, CN, C≡CR, COOR, CONR₂, OOCR, COR, and NO₂, wherein each R is independently H, C1-C8 alkyl, C2-C8 heteroalkyl, C1-C8 acyl, C2-C8 heteroacyl, C2-C8 alkenyl, C2-C8 heteroalkenyl, C2-C8 alkynyl, C2-C8 heteroalkynyl, C3-C8 heterocyclyl, C4-C10 heterocyclylalkyl, C6-C10 aryl, or C5-C10 heteroaryl, and each R is optionally substituted with halo, =O, =N-CN, =N-OR', =NR', OR', NR'₂, SR', SO₂R', SO₂NR'₂, NR'SO₂R', NR'CONR'₂, NR'CSNR'₂, NR'C(=NR')NR'₂, NR'COOR', NR'COR', CN, C≡CR', COOR', CONR'₂, OOCR', COR', and NO₂, wherein each R' is independently H, C1-C8 alkyl, C2-C8 heteroalkyl, C1-C8 acyl, C3-C8 heterocyclyl, C2-C8 heteroacyl, C6-C10 aryl or C5-C10 heteroaryl, and each of the substitutable groups on R' can be substituted with one or more (e.g., up to three) halo, piperidinyl, pyrrolidinyl, piperazinyl, morpholinyl, CN, C1-C4 alkoxy, OH, OAc, NH₂, C1-C4 alkyl amine, di(C1-C4 alkyl)amine, NHAc, NHCOOMe, NHCOOEt, NHCOOtBu, NHSO₂Me, SMe, SO₂Me, SO₂NH₂, SO₂NMe₂, COOH, CONH₂, COOMe, COOEt, CONHMe, or CONMe₂.

"Acetylene" substituents are 2-10C alkynyl groups that contain at least one carbon-carbon triple bond and are optionally substituted with the groups described herein as suitable for alkyl groups; in some embodiments, the alkynyl groups are of the formula -C≡C-R^a, wherein R^a is H or C1-C8 alkyl, C2-C8 heteroalkyl, C2-C8 alkenyl, C2-C8 heteroalkenyl, C2-C8 alkynyl, C2-C8 heteroalkynyl, C1-C8 acyl, C2-C8 heteroacyl, C6-C10 aryl, C5-C10 heteroaryl, C7-C12 arylalkyl, or C6-C12 heteroarylalkyl.

Each R^a group is optionally substituted with one or more substituents selected from halo, =O, =N-CN, =N-OR', =NR', OR', NR'₂, SR', SO₂R', SO₂NR'₂, NR'SO₂R', NR'CONR'₂, NR'CSNR'₂, NR'C(=NR')NR'₂, NR'COOR', NR'COR', CN, COOR', CONR'₂, OOCR', COR', and NO₂, wherein each R' is independently H, C1-C6 alkyl, C2-C6 heteroalkyl, C1-C6 acyl, C2-C6 heteroacyl, C6-C10 aryl, C5-C10 heteroaryl, C7-12 arylalkyl, or C6-12 heteroarylalkyl, each of which is optionally substituted with one or more groups selected from halo, CN, C1-C4 alkyl, C2-C4 heteroalkyl, C1-C6 acyl, C1-C6 heteroacyl, C1-C4 alkoxy, C1-C4 alkylamino, di(C1-C4 alkyl)amino, hydroxy, amino, and =O; and wherein two R' can be linked to form a 3-7 membered ring optionally containing up to three heteroatoms selected from N, O and S. In some embodiments, R^a of -C≡C-R^a is H or Me.

"Heteroalkyl", "heteroalkenyl", and "heteroalkynyl" are defined similarly to the corresponding hydrocarbyl (alkyl, alkenyl and alkynyl) groups, but the 'hetero' terms refer to groups that contain 1-3 O, S or N heteroatoms or combinations thereof within the backbone residue; thus at least one carbon atom of a corresponding alkyl, alkenyl, or alkynyl group is replaced by one of the specified heteroatoms to form, respectively, a heteroalkyl, heteroalkenyl, or heteroalkynyl group. The typical and preferred sizes for heteroforms of alkyl, alkenyl and alkynyl groups are generally the same as for the corresponding hydrocarbyl groups, and the substituents that may be present on the heteroforms are the same as those described above for the hydrocarbyl groups. For reasons of chemical stability, it is also understood that, unless otherwise specified, such groups do not include more than two contiguous heteroatoms except where an oxo group is present on N or S as in a nitro or sulfonyl group.

While "alkyl" as used herein includes cycloalkyl and cycloalkylalkyl groups, the term "cycloalkyl" may be used herein to describe a carbocyclic non-aromatic group that is connected via a ring carbon atom, and "cycloalkylalkyl" may be used to describe a carbocyclic non-aromatic group that is connected to the molecule through an alkyl linker.

Similarly, "heterocyclyl" may be used to describe a non-aromatic cyclic group that contains at least one heteroatom (typically selected from N, O and S) as a ring member and that is connected to the molecule via a ring atom, which may be C (carbon-linked) or N (nitrogen-linked); and "heterocyclylalkyl" may be used to describe such a group that is connected to another molecule through a linker. The heterocyclyl can be fully saturated or partially saturated, but non-aromatic. The sizes and substituents that are suitable for the cycloalkyl, cycloalkylalkyl, heterocyclyl, and heterocyclylalkyl groups are the same as those described above for alkyl groups. The heterocyclyl groups typically contain 1, 2 or 3 heteroatoms, selected from N, O and S as ring members; and the N or S can be substituted with the groups commonly found on these atoms in heterocyclic systems. As used herein, these terms also include rings that contain a double bond or two, as long as the ring that is attached is not aromatic. The substituted cycloalkyl and heterocyclyl groups also include cycloalkyl or heterocyclic rings fused to an aromatic ring or heteroaromatic ring, provided the point of attachment of the group is to the cycloalkyl or heterocyclyl ring rather than to the aromatic / heteroaromatic ring.

Like alkyl groups, the cycloalkyl and heterocyclyl groups described herein can be substituted to the extent permitted by their valence and stability considerations, which are well understood by those of skill in the art. Substituents for the cycloalkyl and heterocyclyl rings or ring systems include those described herein as suitable for placement on alkyl groups.

As used herein, "acyl" encompasses groups comprising an alkyl, alkenyl, alkynyl, aryl or arylalkyl radical attached at one of the two available valence positions of a carbonyl carbon atom, and heteroacyl refers to the corresponding groups wherein at least one carbon other than the carbonyl carbon has been replaced by a heteroatom chosen from N, O and S. Thus heteroacyl includes, for example, -C(=O)OR and -C(=O)NR₂ as well as -C(=O)-heteroaryl.

Acyl and heteroacyl groups are bonded to any group or molecule to which they are attached through the open valence of the carbonyl carbon atom. Typically, they are C1-C8 acyl groups, which include formyl, acetyl, pivaloyl, and benzoyl, and C2-C8 heteroacyl, groups, which include methoxyacetyl, ethoxycarbonyl, and 4-pyridinoyl. The hydrocarbyl groups, aryl groups, and heteroforms of such groups that comprise an acyl or heteroacyl group can be substituted with the substituents described herein as generally suitable substituents for each of the corresponding component of the acyl or heteroacyl, group.

"Aromatic" moiety or "aryl" moiety refers to a monocyclic or fused bicyclic moiety having the well-known characteristics of aromaticity; examples include phenyl and naphthyl. Similarly, "heteroaromatic" and "heteroaryl," refer to such monocyclic or fused bicyclic ring systems which contain as ring members one or more heteroatoms selected from O, S and N. The inclusion of a heteroatom permits aromaticity in 5-membered rings as well as 6-membered rings. Typical heteroaromatic systems include monocyclic C5-C6 aromatic groups such as pyridyl, pyrimidyl, pyrazinyl, thienyl, furanyl, pyrrolyl, pyrazolyl, thiazolyl, oxazolyl, triazolyl, triazinyl, tetrazolyl, tetrazinyl, and imidazolyl and the fused bicyclic moieties formed by fusing one of these monocyclic groups with a phenyl ring or with any of the heteroaromatic monocyclic groups to form a C8-C10 bicyclic group such as indolyl, benzimidazolyl, indazolyl, benzotriazolyl, isoquinolyl, quinolyl, benzothiazolyl, benzofuranyl, pyrazolopyridyl, quinazolinyl, quinoxalinyl and cinnolinyl. Any monocyclic or fused ring bicyclic system which has the characteristics of aromaticity in terms of electron distribution throughout the ring system is included in this definition. It also includes bicyclic groups where at least the ring which is directly attached to the remainder of the molecule has the characteristics of aromaticity. Typically, the ring systems contain 5-12 ring member atoms and up to four heteroatoms selected from N, O and S. Frequently, the monocyclic heteroaryls contain 5-6 ring members and up to three such heteroatoms, and the bicyclic heteroaryls contain 8-10 ring members and up to four such heteroatoms. The number and placement of heteroatoms in such rings is in accordance with the well-known limitations of aromaticity and stability, where stability requires the heteroaromatic group to be stable enough to be exposed to water without rapid degradation.

Aryl and heteroaryl moieties may be substituted with a variety of substituents including C1-C8 alkyl, C2-C8 alkenyl, C2-C8 alkynyl, C5-C12 aryl, C1-C8 acyl, and heteroforms of these, each of which can itself be further substituted; other substituents for aryl and heteroaryl moieties include halo, OR, NR₂, SR, SO₂R, SO₂NR₂, NRSO₂R, NRCONR₂, NRCSNR₂, NRC(=NR)NR₂, NRCOOR, NRCOR, CN, C≡CR, COOR, CONR₂, OOCR, COR, and NO₂, wherein each R is independently H, C1-C8 alkyl, C2-C8 heteroalkyl, C2-C8 alkenyl, C2-C8 heteroalkenyl, C2-C8 alkynyl, C2-C8 heteroalkynyl, C3-C8 heterocyclyl, C4-C10 heterocyclylalkyl, C6-C10 aryl, C5-C10 heteroaryl, C7-C12 arylalkyl, or C6-C12 heteroarylalkyl, and each R is optionally substituted as described above for alkyl groups. The substituent groups on an aryl or heteroaryl group may of course be further substituted with the groups described herein as suitable for each type of such substituents or for each component of the substituent. Thus, for example, an arylalkyl substituent may be substituted on the aryl portion with substituents described herein as typical for aryl groups, and it may be further substituted on the alkyl portion with substituents described herein as typical or suitable for alkyl groups. Where a substituent group contains two R groups on the same or adjacent atoms (e.g.,-NR₂, or -NR-C(O)R), the two R groups can optionally be taken together with the atom(s) in the substituent group to which the are attached to form a ring having 5-8 ring members, which can be substituted as allowed for the R itself, and can contain an additional heteroatom (N, O or S) as a ring member.

Similarly, "arylalkyl" and "heteroarylalkyl" refer to aromatic and heteroaromatic ring systems which are bonded to their attachment point through a linking group such as an alkylene, including substituted or unsubstituted, saturated or unsaturated, cyclic or acyclic linkers. Typically the linker is C1-C8 alkyl or a hetero form thereof. These linkers may also include a carbonyl group, thus making them able to provide substituents as an acyl or heteroacyl moiety. An aryl or heteroaryl ring in an arylalkyl or heteroarylalkyl group may be substituted with the same substituents described above for aryl groups. Preferably, an arylalkyl group includes a phenyl ring optionally substituted with the groups defined above for aryl groups and a C1-C4 alkylene that is unsubstituted or is substituted with one or two C1-C4 alkyl groups or heteroalkyl groups, where the alkyl or heteroalkyl groups can optionally cyclize to form a ring such as cyclopropane, dioxolane, or oxacyclopentane. Similarly, a heteroarylalkyl group preferably includes a C5-C6 monocyclic heteroaryl group that is optionally substituted with the groups described above as substituents typical on aryl groups and a C1-C4 alkylene that is unsubstituted or is substituted with one or two C1-C4 alkyl groups or heteroalkyl groups, or it includes an optionally substituted C5-C6 monocyclic heteroaryl and a C1-C4 heteroalkylene that is unsubstituted or is substituted with one or two C1-C4 alkyl or heteroalkyl groups, where the alkyl or heteroalkyl groups can optionally cyclize to form a ring such as cyclopropane, dioxolane, or oxacyclopentane.

Where an arylalkyl or heteroarylalkyl group is described as optionally substituted, the substituents may be on either the alkyl or heteroalkyl portion or on the aryl or heteroaryl portion of the group. The substituents optionally present on the alkyl or heteroalkyl portion are the same as those described above for alkyl groups generally; the substituents optionally present on the aryl or heteroaryl portion are the same as those described above for aryl groups generally.

"Arylalkyl" groups as used herein are hydrocarbyl groups if they are unsubstituted, and are described by the total number of carbon atoms in the ring and alkylene or similar linker. Thus a benzyl group is a C7-arylalkyl group, and phenylethyl is a C8-arylalkyl.

"Heteroarylalkyl" as described above refers to a moiety comprising an aryl group that is attached through a linking group, and differs from "arylalkyl" in that at least one ring atom of the aryl moiety or one atom in the linking group is a heteroatom selected from N, O and S. The heteroarylalkyl groups are described herein according to the total number of atoms in the ring and linker combined, and they include heteroaryl groups linked through a hydrocarbyl linker such as an alkylene; and heteroaryl groups linked through a heteroalkyl linker. Thus, for example, C7-heteroarylalkyl would include pyridylmethyl and N-pyrrolylmethoxy.

"Alkylene" as used herein refers to a divalent hydrocarbyl group; because it is divalent, it can link two other groups together. Typically it refers to -(CH₂)_n- where n is 1-8 and preferably n is 1-4, though where specified, an alkylene can also be substituted by other groups, and can be of other lengths, and the open valences need not be at opposite ends of a chain. Thus -CH(Me)- and -C(Me)₂- may also be referred to as alkylenes, as can a cyclic group such as cyclopropan-1,1-diyl. Where an alkylene group is substituted, the substituents include those typically present on alkyl groups as described herein.

In general, any alkyl, alkenyl, alkynyl, acyl, or aryl or arylalkyl group or any heteroform of one of these groups that is contained in a substituent may itself optionally be substituted by additional substituents. The nature of these substituents is similar to those recited with regard to the primary substituents themselves if the substituents are not otherwise described. Thus, where an embodiment of, for example, R^x is alkyl, this alkyl may optionally be substituted by the remaining substituents listed as embodiments for R^x where this makes chemical sense, and where this does not undermine the size limit provided for the alkyl per se; e.g., alkyl substituted by alkyl or by alkenyl would simply extend the upper limit of carbon atoms for these embodiments, and is not included. However, alkyl substituted by aryl, amino, alkoxy, =O, and the like would be included within the scope of the invention, and the atoms of these substituent groups are not counted in the number used to describe the alkyl, alkenyl, etc. group that is being described. Where no number of substituents is specified, each such alkyl, alkenyl, alkynyl, acyl, or aryl group may be substituted with a number of substituents according to its available valences; in particular, any of these groups may be substituted with fluorine atoms at any or all of its available valences, for example.

"Heteroform" as used herein refers to a derivative of a group such as an alkyl, aryl, or acyl, wherein at least one carbon atom of the designated carbocyclic group has been replaced by a heteroatom selected from N, O and S. Thus the heteroforms of alkyl, alkenyl, alkynyl, acyl, aryl, and arylalkyl are heteroalkyl, heteroalkenyl, heteroalkynyl, heteroacyl, heteroaryl, and heteroarylalkyl, respectively. It is understood that no more than two N, O or S atoms are ordinarily connected sequentially, except where an oxo group is attached to N or S to form a nitro or sulfonyl group.

"Halo", as used herein includes fluoro, chloro, bromo and iodo. Fluoro and chloro are often preferred.

"Amino" as used herein refers to NH₂, but where an amino is described as "substituted" or "optionally substituted", the term includes NR'R" wherein each R' and R" is independently H, or is an alkyl, alkenyl, alkynyl, acyl, aryl, or arylalkyl group or a heteroform of one of these groups, and each of the alkyl, alkenyl, alkynyl, acyl, aryl, or arylalkyl groups or heteroforms of one of these groups is optionally substituted with the substituents described herein as suitable for the corresponding group. The term also includes forms wherein R' and R" are taken together with the N to which they are attached to form a 3-8 membered ring which may be saturated, unsaturated or aromatic and which contains 1-3 heteroatoms independently selected from N, O and S as ring members, and which is optionally substituted with the substituents described as suitable for alkyl groups or, if NR'R" is an aromatic group, it is optionally substituted with the substituents described as typical for heteroaryl groups.

As used herein, the term "carbocycle", "carbocyclyl", or "carbocyclic" refers to a cyclic ring containing only carbon atoms in the ring, whereas the term "heterocycle" or "heterocyclic" refers to a ring comprising a heteroatom. The carbocyclyl can be fully saturated or partially saturated, but non-aromatic. For example, the carbocyclyl encompasses cycloalkyl. The carbocyclic and heterocyclic structures encompass compounds having monocyclic, bicyclic or multiple ring systems; and such systems may mix aromatic, heterocyclic, and carbocyclic rings. Mixed ring systems are described according to the ring that is attached to the rest of the compound being described; for example, where W represents 1,2,3,4-tetrahydronaphth-1-yl, the group would be encompassed by an optionally substituted cycloalkyl, or carbocyclic group, while the group 1,2,3,4-tetrahydronaphth-6-yl would be included within optionally substituted aromatic groups.

As used herein, the term "heteroatom" refers to any atom that is not carbon or hydrogen, such as nitrogen, oxygen or sulfur. When it is part of the backbone or skeleton of a chain or ring, a heteroatom must be at least divalent, and will typically be selected from N, O, P, and S.

Illustrative examples of heterocycles and heteroaryls include but are not limited to tetrahydrofuran, 1,3-dioxolane, 2,3-dihydrofuran, pyran, tetrahydropyran, benzofuran, isobenzofuran, 1,3-dihydro-isobenzofuran, isoxazole, 4,5-dihydroisoxazole, piperidine, pyrrolidine, pyrrolidin-2-one, pyrrole, pyridine, pyrimidine, octahydro-pyrrolo[3,4 b]pyridine, piperazine, pyrazine, morpholine, thiomorpholine, imidazole, imidazolidine 2,4-dione, 1,3-dihydrobenzimidazol-2-one, indole, thiazole, benzothiazole, thiadiazole, thiophene, tetrahydro thiophene 1,1-dioxide, diazepine, triazole, guanidine, diazabicyclo[2.2.1]heptane, 2,5-diazabicyclo[2.2.1]heptane, 2,3,4,4a,9,9a-hexahydro-1H-β-carboline, oxirane, oxetane, tetrahydropyran, dioxane, lactones, aziridine, azetidine, piperidine, lactams, and may also encompass heteroaryls. Other illustrative examples of heteroaryls include but are not limited to furan, pyrrole, pyridine, pyrimidine, imidazole, benzimidazole and triazole.

The compounds of the invention have structural Formula (II) or (II') as shown above (including IIa, IIa', IIb, IIb', II-TH, and II-TH'). These compounds are typically more selective for CK2, and are highly potent on CK2.

In one embodiment of Formula (II) or (II'), the optionally substituted carbocyclyl is an optionally substituted C3-C8 cycloalkyl; the optionally substituted carbocyclylalkyl is an optionally substituted C4-C10 cycloalkylalkyl; and the optionally substituted heteroalkyl is an optionally substituted C1-C6 alkoxy, optionally substituted C1-C6 alkylamino, or optionally substituted C1-C6 dialkylamino.

In one embodiment of Formula (II) or (II'), -L-W is -NHR⁷, -OR⁷, or -S(O)_nR⁷; n is 0, 1, or 2; and R⁷ is optionally substituted C1-C10 alkyl, optionally substituted heteroalkyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted arylalkyl, optionally substituted heteroarylalkyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted carbocyclylalkyl, or optionally substituted heterocyclylalkyl.

In one embodiment of Formula (II) or (II'), -L-W is -NR⁷R⁸; and R⁷ and R⁸, taken together with the nitrogen atom to which they are attached, form an optionally substituted hetercyclyl which optionally contains one or more additional heteroatom as ring members.

In one embodiment of Formula (II) or (II'), -L-W is optionally substituted aryl, optionally substituted heteroaryl, optionally substituted carbocycyl, or optionally substituted heterocyclyl.

In one embodiment of Formula (II) or (II'), R^1A and R^1B are independently selected from H, optionally substituted C1-C10 alkyl, optionally substituted heterocyclyl, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted heterocyclylalkyl, optionally substituted arylalkyl, or an optionally substituted 5-6 membered aryl ring containing up to two heteroatoms as ring members. Preferably the amine group - NR^1AR^1B in compounds of Formulas (II) or (IIa) and II' or IIa' is not -NH₂, -NHMe, or -NMe₂.

Suitably, R^1A can be selected from H, C1-C4 alkyl, and C1-C6 acyl, where the alkyl and acyl are optionally substituted. In many embodiments, R^1A is H; in other embodiments, it is sometimes Me, or an optionally substituted C1-C4 alkyl. In some embodiments, R^1A is an optionally substituted C1-C6 acyl group, particularly one that can readily be cleaved under mild conditions, such as methoxyacetyl, hydroxyacetyl, or an alpha-amino acyl group, which can act as pro-drugs for the compounds where R^1A is H.

Often, R^1A in this amine group -NR^1AR^1B is H, and R^1B is a substituted or unsubstituted group selected from C2-C8 alkyl, C3-C8 cycloalkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, heterocyclyl, and heterocyclylalkyl. Typically, this aryl is phenyl; heteroaryl refers to a 5-6 membered ring containing up to three heteroatoms selected from N, O and S as ring members; and heterocyclyl refers to a 3-8 membered ring containing at least one heteroatom, and optionally two heteroatoms for 6-8 membered rings, as ring members, where the heteroatoms are selected from N, O and S; and the -alkyl- versions of these (arylalkyl, heteroarylalkyl, and heterocyclylalkyl) typically comprise the specified cyclic group linked via an alkylene linker such as (CH₂)_1-4 to the nitrogen atom of NR^1AR^1B. In certain embodiments, R^1B comprises at least one ring having 3-8 ring members.

Examples of suitable R^1B groups include ethyl, isopropyl, t-butyl, cyclopropyl, cyclobutyl, cyclopentyl, tetrahydrofuranyl, piperidinyl, pyrrolidinyl, cyclopropylmethyl, cyclobutylmethyl, phenyl, and the like, each of which can be unsubstituted or substituted with up to three substituents. Some preferred embodiments include cyclopropyl, isopropyl, t-butyl, and cyclobutyl.

Specific examples of substituted R^1B groups include 2,2,2-trifluoroethyl, 2-methoxyethyl, 2-ethoxyethyl, methoxymethyl, 2-aminoethyl, 2-(N-morpholino)ethyl, 3-hydroxypropyl, 3-dimethylaminopropyl, 3-methoxypropyl, 2-hydroxyethyl, 2-hydroxypropyl, acetyl, benzoyl, phenyl substituted with -COOH, -COOMe, -COOEt, -CONH₂, -CONMe₂, and

• where Q represents a functional group such as -OH, -OR, -COOH, -COOR, -NH₂, -NHR, -NR₂, -CONH₂, -CONHR, -CONR₂, -SR, -S(O)R, -SO₂R, -SONR₂, -C(O)R, -NRC(O)R, -NRC(O)OR, -OC(O)OR, -OC(O)NR₂,
• wherein each R is independently H or an optionally substituted C1-C4 alkyl, group, and two R present on the same functional group can be taken together to form a 5-8 membered optionally substituted ring, which can contain up to two heteroatoms selected from N, O and S as ring members.

Where one or more substituents are present on these R, R^1A, or R^1B groups, often the substituents are selected from halo, OR", N(R")₂, S(O)_mR", COOR", CON(R")₂, CN, phenyl, pyridinyl, pyrrolidinyl, and the like, where each R" is independently selected from H and C1-C4 alkyl, optionally substituted with one or more groups selected from OH, C1-C4 alkoxy, halo, NH₂, C1-C4 alkylamine, and di(C1-C4)alkyl amine, and piperidine, pyrrolidine, morpholine, or furan; and m is 0-2. Frequently, R^1B comprises at least one ring, such as a heterocyclyl or cycloalkyl or aryl ring. A preferred embodiment of R^1B in the amine group -NR^1AR^1B in Formulas (II) and (II') is cyclopropyl, and a preferred embodiment of R^1A is H.

In compounds of Formula (II) and (II') and (IIa) or (IIa'), L can be a bond, - CR⁷=CR⁸-, -C≡C-, -NR⁷-, -O-, -S(O)_n-, or (CR⁷R⁸)_n, or it can be -(CR⁷R⁸)_m-NR⁷-, -(CR⁷R⁸)_m-O-, or -(CR⁷R⁸)_m-S(O)_n-. Typically, where L is attached to W at a heteroatom of W, L will be a bond or one of the hydrocarbon linkers, such as (CR⁷R⁸)_m. However, embodiments of the invention include compounds wherein -L-W is a group of the formula -NR⁷-NR⁷R⁸ as well. Some examples of suitable groups for L include -CH=CH-, -C=C-, -NH-, NMe, -O-, -S-, - S(O)₂-, and -CH₂NH-. Where L is attached to W at a heteroatom of W, L is often CH₂ or (CH₂)₂.

Figures 1 and 2 illustrate the improved selectivity found for compounds of Formula II. Figure 1 depicts a prior art compound of Formula I that is a potent inhibitor of CK2. In assays for inhibition of a panel of 108 kinases, this compound at a comcentration of 1 micromolar is a potent inhibitor of many of the various kinases. By comparison, Figure 2 shows a similar compound of Formula II, having a substituted amine group as an additional substituent on the six-membered ring of the bicyclic core. This compound is more potent as an inhibitor of CK2 than the similar-looking compound in Figure 1; it is less potent as an inhibitor of PIM1; and as the kinase panel assay shows, it is less potent on many other kinases than the compound of Figure 1 is. Relatively few kinases are inhibited by more than 80% with the amine-substituted compound of Formula II, when compared to the proportion of kinase inhibitors showing similar levels of inhibition by the non-aminated compound of Formula I. This improved selectivity is observed for a wide array of amine substituent groups, as the data in Tables 1 and 2 and additional data throughout the application demonstrate.

Specific embodiments of the compounds of the invention include compounds of Formula IIa and/or IIa': wherein,

• R² is H, CH₃ or CF₃;
• Z³ and Z⁴ each independently represent N or CR⁵, or CH;
• where each R⁵ is independently selected from halo, -CN, -R, -OR, -S(O)_nR, -COOR,-CONR², and -NR₂,
• wherein each R is independently selected from H and optionally substituted C1-C4 alkyl, or the two R groups, taken together with the nitrogen atom to which they are attached, form an optionally substituted 5- or 6-membered heterocyclic ring which contains one or more additional heteroatom selected from N, O and S as a ring member;
• R⁴ is H, CH₃ or CF₃;
• X is O, S or NH;
• Y is O or S;
• R^1B is selected from H, optionally substituted C1-C10 alkyl, optionally substituted heteroalkyl, optionally substituted heterocyclyl, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted heterocyclylalkyl, optionally substituted arylalkyl, or an optionally substituted heteroaryl;
• L is a bond, -NR⁷-, -O-, -S(O)_n-, (CR⁷R⁸)_m, or -(CR⁷R⁸)_m-NR⁷-;
• m is 1, 2, 3, or 4;
• n is 0, 1, or 2;
• W is selected from optionally substituted aryl, optionally substituted heteroaryl, and -NR⁷R⁸,
• where each R⁷ and R⁸ is independently selected from H, optionally substituted C1-C6 alkoxy, optionally substituted C1-C6 alkylamino, optionally substituted C1-C6 dialkylamino, optionally substituted heterocyclyl, optionally substituted C1-C10 alkyl, optionally substituted C3-C8 cycloalkyl, optionally substituted C4-C10 cycloalkylalkyl, optionally substituted aryl, optionally substituted arylalkyl, optionally substituted heteroaryl, and optionally substituted heteroarylalkyl;
• and R⁷ and R⁸, taken together on a single carbon atom or on adjacent connected carbon atoms of (CR⁷R⁸)_m whether alone or as part of another group, form a 3- to 8-membered ring that contains one or more heteroatoms as ring members;
• or R⁷ and R⁸, taken together with the nitrogen atom to which they are attached, form an optionally substituted 5- to 10-membered heterocyclic or heteroaryl ring system that optionally contains an additional heteroatom selected from N, O and S as a ring member; and
• provided that no more than one of or R⁷ and R⁸ in -NR⁷R⁸ is selected from the group consisting of alkoxy, alkylamino, dialkylamino and heterocyclyl.

In the foregoing compounds of Formula (IIa) or (IIa'), R² and R⁴ are selected from H, CH₃ and CF₃. In some embodiments R² is H. In some embodiments, R⁴ is H.

In the foregoing compounds of Formula (IIa) or (IIa'), Y is O or S. In preferred embodiments, Y is O.

In the foregoing compounds of Formula (IIa) or (IIa'), X can be S, O or NH. Frequently, X is NH or S. In certain embodiments, X is NH.

In the foregoing compounds of Formula (IIa) or (IIa'), Z³ and Z⁴ are often selected from N and CH. In some embodiments, one of these ring members is N and the other is CH. In alternative embodiments, both Z³ and Z⁴ are N. In still other embodiments, Z³ and Z⁴ are both CH.

In certain compounds of Formula IIa, Z³ can be N while Z⁴ is CH; or Z³ can be N while Z⁴ is also N. In certain compounds of Formula IIa', Z³ can be CH while Z⁴ is N; alternatively, Z³ can be N while Z⁴ is N or CH.

In the foregoing compounds of Formula (IIa) or (IIa'), R³, when present, can be H or optionally substituted alkyl. Often, R³ is H.

Z can be O or S; in preferred embodiments, Z is O.

When present, m is frequently 1 or 2.

In these compounds of Formula IIa and/or IIa', R² and R⁴ are frequently both H.

In the foregoing compounds of Formula IIa and/or IIa', R^1B can be optionally substituted C1-C10 alkyl, optionally substituted heterocyclyl, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted heterocyclylalkyl, optionally substituted arylalkyl, or an optionally substituted 5-6 membered aryl ring containing up to two heteroatoms as ring members. In some embodiments, R^1B is a C3-C6 cycloalkyl, or a 3-6 membered heterocyclic group such as piperidine or a C1-C3 alkyl group substituted with one of these rings, and it is optionally substituted. Specific embodiments of R^1B include cyclopropyl, cyclopropylmethyl, 4-piperidinyl, and substituted 4-piperidinyl, e.g. 4-piperidinyl substituted with an acyl group, such as acetyl, at N-1. Other embodiments include optionally substituted phenyl.

In the foregoing compounds of Formula (IIa) or (IIa'), -L-M is -NHR⁷, -OR⁷, or-S(O)_nR⁷; n is 0, 1, or 2; and R⁷ is optionally substituted C1-C10 alkyl, optionally substituted heteroalkyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted arylalkyl, optionally substituted heteroarylalkyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted carbocyclylalkyl, or optionally substituted heterocyclylalkyl.

In the foregoing compounds of Formula (IIa) or (IIa'), -L-M is -NR⁷R⁸; and R⁷ and R⁸, taken together with the nitrogen atom to which they are attached, form an optionally substituted heterocyclyl which optionally contains one or more additional heteroatom as ring members.

In the foregoing compounds of Formula (IIa) or (IIa'), -L-M is optionally substituted aryl, optionally substituted heteroaryl, optionally substituted carbocycyl, or optionally substituted heterocyclyl.

In the foregoing compounds of Formula IIa and/or IIa', L is typically a bond or NH. When L is NH, W can be an optionally substituted group selected from phenyl, phenylalkyl, heterocyclyl, cycloalkyl and cycloalkylalkyl.

In the foregoing compounds of Formula IIa and/or IIa', W is frequently an optionally substituted phenyl, arylalkyl, cycloalkyl, heteroaryl, cycloalkylalkyl, or heterocyclic group. Specific examples include optionally substituted phenyl; optionally substituted phenylmethyl; optionally substituted 1-phenylethyl; cyclopropylmethyl; 1-cyclopropylethyl; piperidinyl; and morpholinyl. Some preferred substituents for the pheyl groups of W include halo, CN, Me, CF₃, OMe, OCF₃, and heteroaryl groups such as pyrazole or pyrrole or imidazole.

When L is a bond, W is frequently an optionally substituted aryl, heteroaryl or heterocyclyl group. Surprisingly high flexibility has been demonstrated among the groups that can be represented by W in Formula II, II', and (IIa) or (IIa'). Aryl and heteroaryl groups are suitable for W, and can be unsubstituted or substituted. Examples of suitable aromatic groups include phenyl, pyridinyl, pyrimidinyl, thienyl (thiophene ring), furanyl, oxazole, isoxazole, thiazole, isothiazole, oxadiazole, thiadiazole, triazole, and the like, as well as indole, benzimidazole, benzofuran, benzopyrazole, imidazole, pyrrole, pyrazole, and the like. Note that the latter group (indole, benzimidazole, benzofuran, benzopyrazole, imidazole, pyrrole, pyrazole) contain a 5-membered nitrogen heterocycle, and can be linked to L through either C or N as a result. In some embodiments, W represents one of these aromatic groups that comprises a 5-membered ring, and W is attached via N of the 5-membered ring to L, and L is a bond so that W is effectively attached directly to the ring containing Z³ and Z⁴. Suitable substituents for all of these aryl or heteroaryl groups include those described herein as suitable for such aromatic groups.

When W is an aromatic group, L is sometimes a bond, NH, or O. A particular embodiment of interest is a compound of Formula II, II', (IIa) or (IIa'), wherein L is a bond or NH, and W is an optionally substituted phenyl or optionally substituted thienyl ring. In embodiments where L is a bond, it is often desirable for the position of each ring atom of the aryl ring that is adjacent to the attachment point for L to be unsubstituted (i.e., any adjacent carbon(s) would be CH), so the optional substituents on W in such compounds are often, when present, located at positions 3, 4, or 5 of a phenyl ring (assuming position 1 attaches to L), or to positions 4 or 5 of a thienyl ring when L attaches to position 2, and at position 5 of the thienyl group when L attaches at position 3. Examples of these W groups include: where each A represents the presence of an optional substituent (or more than one where the ring valence permits more) on a carbon not having an explicit H attached.

Where W is an aromatic group, a wide array of substituents are well tolerated and provide high levels of kinase activity. Suitable substituents include those described herein as suitable for placement on aromatic groups in general. Some of the suitable substituents for these aromatic group W's include halo (especially F or Cl), alkyl, (e.g., C1-C4 alkyl, such as methyl, ethyl, isopropyl or cyclopropyl); alkoxy (especially C1-C4 alkyloxy); haloalkyl (e.g., CF³, -CH₂CF₃); haloalkoxy (e.g. -OCF₃, -OCF₂H, OCH₂CF₃, and the like); CN, -OH, alkynyl (e.g., -CCH, CCMe, and the like); heterocyclylmethyl (e.g., N-piperidinylmethyl. N-pyrrolidinylmethyl, N-morpholinylmethyl, etc.); hydroxymethyl, aminomethyl, dimethylaminomethyl, methylaminomethyl; substituted C1-C4 alkoxy such as methoxyethoxy, ethoxymethoxy, trifluoroethoxy, 2-(N-morpholino)ethoxy, 2-(N-pyrrolidinyl)ethoxy, 2-(piperidinyl)ethoxy, and the like; acyl groups of the formula -C(O)-X, where X represents -OR, -NR₂, or -R, where each R is independently selected from H or an optionally substituted member selected from C1-C4 alkyl, 3-8 membered cycloalkyl or heterocyclyl, and 5-6 membered aryl or heteroaryl containing up to 3 heteroatoms'selected from N, O and S as ring members, and where two R on one group (e.g., two R's of -NR₂) can be taken together to form an optionally substituted 5-8 membered ring containing up to two heteroatoms selected from N, O and S as ring members; heterocyclic groups such as morpholine, tetrahydrofuran, piperidine, pyrrolidine, 4-Me-N-piperazinyl, N-piperazinyl, 4-acetyl-N-piperazinyl, and the like.

Commonly, an aromatic group W will have 1-2 substituents, or it will be unsubstituted; and commonly the substituents, when present, are positioned as described above, so that the ring carbon(s) adjacent to where L is attached are unsubstituted (CH). When L is other than a bond, the substituents on W can be at any position, and often will be at the positions ortho and/or para to the point of attachment of W to L.

Alternatively, W can be a heterocyclic group such as piperidinyl, morpholinyl, pyrrolidinyl, tetrahydrofuranyl, tetrahydropyranyl, thiomorpholinyl, piperazinyl, thiolanyl, and the like, each of which can be unsubstituted or substituted with up to four substituents. Suitable substituents for these groups include those described herein as suitable for heterocyclic groups. Note that even when L is NR or NH, W can be a heterocyclic groups such as 1-piperidinyl or 4-morpholinyl where L links to a heteroatom (N) of the heterocyclic group as well as at C of the heterocyclic group.

Where L is NH, W can also be arylalkyl, or cycloalkylalkyl or heterocyclylalkyl, and the alkyl portion of W can be e.g. C1-C4. Where L comprises an alkyl portion, it can be a straight chain (e.g., ethylene, propylene, butylene), or it can be a substituted alkylene chain, resulting in formation of a potentially chiral carbon linker. Where L is a chiral group of this type, e.g. when L is -CH(R)- or -CH₂-CH(R)- where R is not H (e.g., R is Methyl or ethyl, L can be either in an R configuration or an S configuration, where those terms are used in their conventional stereochemical sense, or it can be present as a mixture of isomers, including a racemic mixture. In some embodiments, such a chiral center present in L will be in the S configuration. In other embodiments, it can be in the R configuration.

Alternatively, W can be a group of the formula or -NR⁷R⁸, -OR⁷, S(O)_nR⁷, CONR⁷R⁸, or CR⁷R⁸R⁹, where each R⁷ and R⁸ and R⁹ is independently selected from H, optionally substituted C1-C10 alkyl, optionally substituted aryl, optionally substituted arylalkyl, optionally substituted heteroaryl, and optionally substituted heteroarylalkyl; or R⁷ and R⁸ taken together with the N of -NR⁷R⁸ can form an optionally substituted 5-10 membered heterocyclic or heteroaromatic ring system that optionally contains an additional heteroatom selected from N, O and S as a ring member.

In embodiments where W is -NR⁷R⁸, L is frequently a bond, and R⁷ and R⁸ taken together with the N of NR⁷R⁸ can form an optionally substituted 5-10 membered heterocyclic or heteroaromatic ring system that optionally contains an additional heteroatom selected from N, O and S as a ring member. Suitable such rings include e.g., pyrrolidinyl, piperidinyl, piperazinyl, thiomorpholinyl, diazepinyl, and morpholinyl, each of which can be substituted to the extent substitution forms relatively water-stable structures. Suitable substituents include, for example, oxo (=O), C1-C4 alkyl, -OH, -CN, halo (especially F or Cl), COOR, CONR₂, SR, -S(O)R, -SO₂R, -NR₂, hydroxyalkyl, -OR, methoxyalkyl (e.g., methoxymethyl), where each R is independently H or optionally substituted C1-C4 alkyl, and where two R on one group can be taken together to form an optionally substituted 5-8 membered ring containing up to two heteroatoms selected from N, O and S as ring members.

In some embodiments of the compounds of Formula (II) and (II') and (IIa) or (IIa'),-L-W is a group of the formula -NH-Ar, where Ar represents an optionally substituted aromatic group. Suitable aromatic rings for this group include phenyl, naphthyl, pyridinyl, pyrimidinyl, thienyl (thiophene ring), furanyl, indolyl, benzofuranyl, benzothienyl, benzopyrazolyl, benzimidazolyl, benzoxazole and benzothiazole. Suitable substituents for these aryl or heteroaryl groups include those described herein as suitable for such aromatic groups.

In some embodiments, W is an optionally substituted cycloalkyl group, typically containing 3-8 ring atoms in a monocyclic structure, or 8-10 ring atoms in a bicyclic structure. Examples include 1,2,3,4-tetrahydronaphth-1-yl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and decalin. These groups are optionally substituted as described herein; in some embodiments, the cycloalkyl ring will be substituted with one or more (e.g., up to three) groups selected from halo, hydroxy, oxo (=O), COOR, CONR₂, SR, -S(O)R, -SO₂R, -NR₂, hydroxyalkyl, -OR, methoxyalkyl (e.g., methoxymethyl), C1-C4 alkyl, where each R is independently H or optionally substituted C1-C4 alkyl, and where two R on one group can be taken together to form an optionally substituted 5-8 membered ring containing up to two heteroatoms selected from N, O and S as ring members.

Particular embodiments of the compounds of the invention include thiophene-containing compounds of Formula (II-Th) and (II-Th'):

• where R^Th is selected from H, halo, optionally substituted C1-C6 alkyl, CN, S(O)_0-2R,-SO₂NR₂, COOR, CONR₂, and C(O)R,
• where each R is independently H, halo, CN, or an optionally substituted member selected from the group consisting of C1-C6 alkyl, C1-C6 alkoxy, C1-C6 alkylamino, di(C1-C6)alkylamino, C3-C8 cycloalkyl, C4-C10 cycloalkylalkyl, C5-C8 heterocyclyl, C6-C10 heterocyclylalkyl, aryl, arylalkyl, C5-C6 heteroalkyl, and C6-C10 heteroalkylalkyl;
  • and two R on the same atom or adjacent connected atoms can form an optionally substituted heterocyclic ring that can contain an additional heteroatom selected from N, O and S as a ring member:
• and other structural features are as defined for Formula IIa above.

The thienyl (thiophene) ring in Formulas II-Th and II-Th' can be attached to the bicyclic core at either position 2 or position 3 of the thiophene ring, when the position substituted with R^Th is defined as position 5, and the ring sulfur is position 1. In some embodiments, connection is at position 2 of the thienyl group, and in alternative embodiments, connection is at position 3 of the thienyl group.

In these compounds of Formulas II-Th and II-Th', R² and R⁴ are frequently both H.

In the foregoing compounds of Formulas II-Th and II-Th', X is preferably NH.

In the foregoing compounds of Formulas II-Th and II-Th', Y is frequently O.

In the foregoing compounds of Formulas II-Th and II-Th', Z³ is often N.

In the foregoing compounds of Formulas II-Th and II-Th', Z⁴ can be CH or N.

In the foregoing compounds of Formulas II-Th and II-Th', R^1B can be optionally substituted C1-C10 alkyl, optionally substituted heterocyclyl, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted heterocyclylalkyl, optionally substituted arylalkyl, or an optionally substituted 5-6 membered aryl ring containing up to two heteroatoms as ring members. In some embodiments, R^1B is a C3-C6 cycloalkyl or a 3-6 membered heterocyclic group such as piperidine or a C1-C3 alkyl, group substituted with one of these rings, and it is optionally substituted. Specific embodiments of R^1B include cyclopropyl, cyclopropylmethyl, 4-piperidinyl, and substituted 4-piperidinyl, e.g. 4-piperidinyl substituted with an acyl group, such as acetyl, at N-1. Other embodiments include optionally substituted phenyl.

In these compounds, R^TH can be halo (F, Cl, Br), CF₃, CN, C1-C6 alkyl, C1-C3 alkyl, substituted with heterocyclyl or heterocyclylamino, COOR, or COONR₂.

In one emobidment of the present invention, the compounds of Formula (IIa) or (IIa') have structural Formula (IIb) or (IIb'): wherein

• R² and R⁴ are independently H, CH₃ or CF₃;
• Z⁴ is N or CH;
• -L-M is -NR^8AR⁷, -NHR⁷, -OR⁷, or S(O)_nR⁷;
• n is 0, 1, or 2; and
• R⁷ is optionally substituted C1-C10 alkyl, optionally substituted heteroalkyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted arylalkyl, optionally substituted heteroarylalkyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted carbocyclylalkyl, or optionally substituted heterocyclylalkyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted carbocycyl, or optionally substituted heterocyclyl; or
• R⁷ and R^8A, taken together with the nitrogen atom to which they are attached, form an optionally substituted heterocyclyl which optionally contains one or more additional heteroatom as ring members.

In one emobidment of the present invention, the compounds of Formula (II) have structural Formula (1Ic): wherein,

• X is O, S, or NR²;
• R³ is -(CH₂)-X^C;
• X^C is hydroxyl or a group having structural formula (a), (b), (c), or (d):
  • L¹ and L² are each independently a covalent bond, -O-, or -NR^3a-;
  • R^1a and R^2a are each independently hydrogen, alkyl, heteroalkyl, heteroaryl, heterocyclyl, alkenyl, alkynyl, arylalkyl, heteroarylalkyl, heterocyclylalkyl, -alkylene-C(O)-O-R^4a, or -alkylene-O-C(O)-O-R^4a; and
  • R^3a and R^4a are each independently hydrogen, alkyl, heteroalkyl, cyclylalkyl, heterocyclyl, aryl, heteroaryl, alkenyl, alkynyl, arylalkyl, heterocyclylalkyl, or heteroarylalkyl;
• L³ is a covalent bond or alkylene;
• Y is OR^5a, NR^5aR^6a, or C(O)OR^7a, provided that when Y is C(O)OR^7a, then L³ is not a covalent bond; and
• R^5a, R^6a, and R^7a are each independently hydrogen, alkyl, arylalkyl, aryl, heteroalkyl, alkylheteroaryl, heterocyclyl, or heteroaryl; or alternatively, R^5a and R^6a, taken together with the nitrogen atom to which they are attached, form a hetercyclyl ring optionally containing one o r more additional heteroatom independently selected from N, O, and S.

In one embodiment of Formula (IIc), X is NR²; R³ is -(CH₂)-X^C; and X^C is hydroxyl or a group having structural formula (b):

In one embodiment of Formula (IIc), R² and R⁴ are hydrogen.

In one embodiment of Formula (IIc), R^1B is an optionally substituted C1-C10 alkyl, cycloalkyl, or cycloalkylalkyl,

In one embodiment of Formula (IIc), -L-W is OR⁷ or -NR⁷R⁸.

In one embodiment of Formula (IIc), R⁷ is optionally substituted aryl or optionally substituted heteroaryl; and R⁸ is H.

In one embodiment of Formula (IIc), R⁸ is optionally substituted phenyl.

In one embodiment of Formula (IIc), L³ is a covalent bond; and Y is OR^5a or NR^5aR^6a.

The compounds of the invention also include those enriched in isotopes of the atoms involved in the structures described herein. For example, the compounds as described are intended to include versions wherein one or more H atoms is preferentially enriched in a heavier hydrogen isotope (deuterium or tritium). In particular, where any of the foregoing compounds contains a methyl group (Me), an enriched methyl group containing deuterium at levels far above natural abundance can be used. For example, -CH₃ could be replaced by -CH₂D or-CHD₂ or -CD₃, where each D represents deuterium present in place of ¹H, and indicates that D is present instead of ¹H in at least about 50% of the molecules of a sample of the compound. Of particular interest are compounds comprising -N(R)Me or -NMe₂, where Me can be present as CD₃. This variation of the compounds described herein is particularly interesting because the presence of CD₃ in place of CH₃ can have a significant effect on rates of metabolism of an N-methyl group, thus a compound comprising CD₃ can have improved pharmacokinetic properties over a non-enriched compound. Accordingly, the alkyl groups described herein are intended to include ones enriched in deuterium, and compounds containing a methyl group on N are specifically considered to include a deuterium-enriched methyl group on N.

The compounds of the invention often have ionizable groups so as to be capable of preparation as salts. In that case, wherever reference is made to the compound, it is understood in the art that a pharmaceutically acceptable salt may also be used. These salts may be acid addition salts involving inorganic or organic acids or the salts may, in the case of acidic forms of the compounds of the invention be prepared from inorganic or organic bases. Frequently, the compounds are prepared or used as pharmaceutically acceptable salts prepared as addition products of pharmaceutically acceptable acids or bases. Suitable pharmaceutically acceptable acids and bases are well-known in the art, such as hydrochloric, sulphuric, hydrobromic, acetic, lactic, citric, or tartaric acids for forming acid addition salts, and potassium hydroxide, sodium hydroxide, ammonium hydroxide, caffeine and various amines for forming basic salts. Methods for preparation of the appropriate salts are well-established in the art. In some cases, the compounds may contain both an acidic and a basic functional group, in which case they may have two ionized groups and yet have no net charge.

In another aspect, the invention provides a pharmaceutical composition comprising any of the compounds of the invention as above-described, admixed with a pharmaceutically acceptable excipient.

The compounds of the invention are useful as medicaments, and are useful for the manufacture of medicaments, including medicaments to treat conditions disclosed herein, such as cancers, inflammatory conditions, infections, pain, and immunological disorders.

The terms "treat" and "treating" as used herein refer to ameliorating, alleviating, lessening, and removing symptoms of a disease or condition. A candidate molecule or compound described herein may be in a therapeutically effective amount in a formulation or medicament, which is an amount that can lead to a biological effect, such as apoptosis of certain cells (e.g., cancer cells), reduction of proliferation of certain cells, or lead to ameliorating, alleviating, lessening, or removing symptoms of a disease or condition, for example. The terms also can refer to reducing or stopping a cell proliferation rate (e.g., slowing or halting tumor growth) or reducing the number of proliferating cancer cells (e.g., removing part or all of a tumor).

These terms also are applicable to reducing a titre of a microorganism in a system (i.e., cell, tissue, or subject) infected with a microorganism, reducing the rate of microbial propagation, reducing the number of symptoms or an effect of a symptom associated with the microbial infection, and/or removing detectable amounts of the microbe from the system. Examples of microorganisms include but are not limited to virus, bacterium and fungus.

The compounds of the invention have activities to modulate protein kinases, in particular CK2 activity and/or Pim activity. In some embodiments, the compounds of the invention specifically inhibit the activity of CK2, but not Pim, e.g., more than 100, 90, 80, 70, 60, 50, 40, 30, 20, or 10 fold difference between CK2 inhibition vs. Pim inhibition. In some embodiments, the compounds of the invention specifically inhibit the acitivity of Pim, but not Ck2, e.g., more than 100, 90, 80, 70, 60, 50, 40, 30, 20, or 10 fold difference between Pim inhibition vs. CK2 inhibition. In some embodiments, the compounds of the invention inhibit the activity of CK2 as well as Pim.

The compounds of the invention can be used to modulate the activity of CK2 and/or Prim, e.g., inhibit the activity of CK2 and/or Pim in a cell, e.g., in vivo or in vitro. In some embodiments, compounds of the invention can be used to modulate the activity of CK2, e.g., inhibit the activity of CK2 without substantially interferring or changing the activity of Pim. In some embodiments, compounds of the invention can be used to modulate the activity of Pim, e.g., inhibit the activity of Pim without substantially interferring or changing the activity of CK2. In some embodiments, compounds of the invention can be used to modulate the activity of CK2 and Pim, e.g., inhibit the activity of CK2 and Pim.

The compounds of the invention are thus useful to treat infections by certain pathogens, including protozoans and viruses. The compounds may thus be used in methods for treating protozoal disorders such as protozoan parasitosis, including infection by parasitic protozoa responsible for neurological disorders such as schizophrenia, paranoia, and encephalitis in immunocompromised patients, as well as Chagas' disease. The compounds may also be used to treat various viral diseases, including human immunodeficiency virus type 1 (HIV-1), human papilloma viruses (HPVs), herpes simplex virus (HSV), Epstein-Barr virus (EBV), human cytomegalovirus, hepatitis C and B viruses, influenza virus, Borna disease virus, adenovirus, coxsackievirus, coronavirus and varicella zoster virus. The methods for treating these disorders comprise administering to a subject in need thereof an effective amount of a compound of Formula II or Formula II'.

As used herein, the term "apoptosis" refers to an intrinsic cell self-destruction or suicide program. In response to a triggering stimulus, cells undergo a cascade of events including cell shrinkage, blebbing of cell membranes and chromatic condensation and fragmentation. These events culminate in cell conversion to clusters of membrane-bound particles (apoptotic bodies), which are thereafter engulfed by macrophages.

Compounds of the invention as described above may also be used in methods for identifying a candidate molecule that interacts with a CK2. Such methods comprise contacting a composition containing a CK2 protein and a compound of the invention described herein with a candidate molecule and determining whether the amount of the compound of the invention described herein that interacts with the protein is modulated, whereby a candidate molecule that modulates the amount of the molecule described herein that interacts with the protein is identified as a candidate molecule that interacts with the protein.

Compounds of the invention as described above are also useful for modulating certain protein kinase activities. Protein kinases catalyze the transfer of a gamma phosphate from adenosine triphosphate to a serine or threonine amino acid (serine/threonine protein kinase), tyrosine amino acid (tyrosine protein kinase), tyrosine, serine or threonine (dual specificity protein kinase) or histidine amino acid (histidine protein kinase) in a peptide or protein substrate. A system comprising a protein kinase protein may thus be contacted with a compound of the invention as described herein in an amount effective for modulating (e.g., inhibiting) the activity of the protein kinase. In some embodiments, the activity of the protein kinase is the catalytic activity of the protein (e.g., catalyzing the transfer of a gamma phosphate from adenosine triphosphate to a peptide or protein substrate). In certain embodiments, methods for identifying a candidate molecule that interacts with a protein kinase comprise: contacting a composition containing a protein kinase and a compound-of the invention as described herein with a candidate molecule under conditions in which the compound and the protein kinase interact, and described herein that interacts with the protein kinase is modulated relative to a control interaction between the compound and the protein kinase without the candidate molecule, whereby a candidate molecule that modulates the amount of the compound interacting with the protein kinase relative to the control interaction is identified as a candidate molecule that interacts with the protein kinase. Systems in such embodiments can be a cell-free system or a system comprising cells (e.g., in vitro). The protein kinase, the compound or the molecule in some embodiments is in association with a solid phase. In certain embodiments, the interaction between the compound and the protein kinase is detected via a detectable label, where in some embodiments the protein kinase comprises a detectable label and in certain embodiments the compound comprises a detectable label. The interaction between the compound and the protein kinase sometimes is detected without a detectable label.

Provided also are compositions of matter comprising a protein kinase and a compound of the invention as described herein. In some embodiments, the protein kinase in the composition is a serine-threonine protein kinase. In some embodiments, the protein kinase in the composition is, or contains a subunit (e.g., catalytic subunit, SH2 domain, SH3 domain) of, CK2. In certain embodiments the composition is cell free and sometimes the protein kinase is a recombinant protein.

The protein kinase can be from any source, such as cells from a mammal, ape or human, for example. Examples of serine-threonine protein kinases that can be inhibited, or may potentially be inhibited, by compounds disclosed herein include without limitation human versions of CK2, or CK2α2. A serine-threonine protein kinase sometimes is a member of a subfamily containing one or more of the following amino acids at positions corresponding to those listed in human CK2: leucine at position 45, methionine at position 163 and isoleucine at position 174. Nucleotide and amino acid sequences for protein kinases and reagents are publicly available (e.g., World Wide Web URLs www.ncbi.nlm.nih.gov/sites/entrez/ and www.Invitrogen.com, each last visited December 2, 2009).

Compounds of the invention may also be used in methods for treating a condition related to aberrant cell proliferation. For example, methods of treating a cell proliferative condition in a subject comprise administering a compound of the invention as described herein to a subject in need thereof in an amount effective to treat the cell proliferative condition. The subject may be a research animal (e.g., rodent, dog, cat, monkey), optionally containing a tumor such as a xenograft tumor (e.g., human tumor), for example, or may be a human. A cell proliferative condition sometimes is a tumor, e.g., solid or circulating tumor or non-tumor cancer, including but not limited to, cancers of the colorectum, breast, lung, liver, pancreas, lymph node, colon, prostate, brain, head and neck, skin, liver, kidney, blood and heart (e.g., leukemia, lymphoma, carcinoma).

Compounds and compositions of the invention may be used alone or in combination with anticancer or other agents, such as a palliative agents, that are typically administered to a patient being treated for cancer, as further described herein.

Compounds of the invention are further used in methods for treating a condition related to inflammation or pain. For example, a method for treating pain in a subject comprises administering a compound of the invention described herein to a subject in need thereof in an amount effective to treat the pain. A method of treating inflammation in a subject comprises administering a compound of the invention described herein to a subject in need thereof in an amount effective to treat the inflammation. The subject may be a research animal (e.g., rodent, dog, cat, monkey), for example, or may be a human. Conditions associated with inflammation and pain include without limitation acid reflux, heartburn, acne, allergies and allergen sensitivities, Alzheimer's disease, asthma, atherosclerosis, bronchitis, carditis, celiac disease, chronic pain, Crohn's disease, cirrhosis, colitis, dementia, dermatitis, diabetes, dry eyes, edema, emphysema, eczema, fibromyalgia, gastroenteritis, gingivitis, heart disease, hepatitis, high blood pressure, insulin resistance, interstitial cystitis, joint pain/arthritis/rheumatoid arthritis, metabolic syndrome (syndrome X), myositis, nephritis, obesity, osteopenia, glomerulonephritis (GN), juvenile cystic kidney disease, and type I nephronophthisis (NPHP), osteoporosis, -Parkinson's disease, Guam-Parkinson dementia, supranuclear palsy, Kuf's disease, and Pick's disease, as well as memory impairment, brain ischemia, and schizophrenia, periodontal disease, polyarteritis, polychondritis, psoriasis, scleroderma, sinusitis, Sjögren's syndrome, spastic colon, systemic candidiasis,
tendonitis, urinary tract infections, vaginitis and inflammatory cancer (e.g., inflammatory breast cancer).

Methods for determining and monitoring effects of compounds herein on pain or inflammation are known. For example, formalin-stimulated pain behaviors in research animals can be monitored after administration of a compound described herein to assess treatment of pain (e.g., Li et al., Pain 115(1-2): 182-90 (2005)). Also, modulation of pro-inflammatory molecules (e.g., IL-8, GRO-alpha, MCP-1, TNFalpha and iNOS) can be monitored after administration of a compound described herein to assess treatment of inflammation (e.g., Parhar et al., Int J Colorectal Dis. 22(6): 601-9 (2006)), for example. Thus, methods for determining whether a compound of the invention as described herein reduces inflammation or pain comprises contacting a system with said compound in in an amount effective for modulating (e.g., inhibiting) the activity of a pain signal or inflammation signal.

Methods for identifying a compound that reduces inflammation or pain comprise: contacting a system with a compound of Formula II or Formula II'; and detecting a pain signal or inflammation signal, whereby a compound that modulates the pain signal relative to a control molecule is identified as a compound that reduces inflammation of pain. Non-limiting examples of pain signals are formalin-stimulated pain behaviors and examples of inflammation signals include without limitation a level of a pro-inflammatory molecule. Compounds of the invention may thus be used in methods for modulating angiogenesis in a subject, and methods for treating a condition associated with aberrant angiogenesis in a subject.

CK2 has also been shown to play a role in the pathogenesis of atherosclerosis, and may prevent atherogenesis by maintaining laminar shear stress flow. CK2 plays a role in vascularization, and has been shown to mediate the hypoxia-induced activation of histone deacetylases (HDACs). CK2 is also involved in diseases relating to skeletal muscle and bone tissue, including, e.g., cardiomyocyte hypertrophy, heart failure, impaired insulin signaling and insulin resistance, hypophosphatemia and inadequate bone matrix mineralization.

Thus in one aspect, compounds of the invention may be used to treat each of these conditions. Said treatment comprises administering to a subject in need thereof an effect amount of a CK2 inhibitor, such as a compound of Formula II or Formula II' as described herein.

Compounds of the invention may also be used in methods for modulating an immune response in a subject, and methods for treating a condition associated with an aberrant immune response in a subject. Thus, a method for determining whether a compound of the invention as described herein modulates an immune response comprises contacting a system with a compound of the invention as described herein in an amount effective for modulating (e.g., inhibiting) an immune response or a signal associated with an immune response. Signals associated with immunomodulatory activity include, e.g., stimulation of T-cell proliferation, suppression or induction of cytokines, including, e.g., interleukins, interferon-γ and TNF. Methods of assessing immunomodulatory activity are known in the art.

Methods for treating a condition associated with an aberrant immune response in a subject comprise administering a compound of the invention as described herein to a subject in need thereof in an amount effective to treat the condition. Conditions characterized by an aberrant immune response include without limitation, organ transplant rejection, asthma, autoimmune disorders, including rheumatoid arthritis, multiple sclerosis, myasthenia gravis, systemic lupus erythematosus, scleroderma, polymyositis, mixed connective tissue disease (MCTD), Crohn's disease, and ulcerative colitis. In certain embodiments, an immune response may be modulated by administering a compound herein in combination with a molecule that modulates (e.g., inhibits) the biological activity of an mTOR pathway member or member of a related pathway (e.g., mTOR, PI3 kinase, AKT). In certain embodiments the molecule that modulates the biological activity of an mTOR pathway member or member of a related pathway is rapamycin. In certain embodiments, a compound of the invention described herein may be provided in a composition in combination with a molecule that modulates the biological activity of an mTOR pathway member or member of a related pathway, such as rapamycin.

In another aspect, the invention provides pharmaceutical compositions (i.e., formulations). The pharmaceutical compositions can comprise a compound of any of Formulae (II), (II'), (IIa), (IIa'), (IIb), (IIb'), (II-Th), and (II-Th'), as described herein which is admixed with at least one pharmaceutically acceptable excipient or carrier. Frequently, the composition comprises at least two pharmaceutically acceptable excipients or carriers.

While the compositions of the present invention will typically be used in therapy for human patients, they may also be used in veterinary medicine to treat similar or identical diseases. The compositions may, for example, be used to treat mammals, including, but not limited to, primates and domesticated mammals. The compositions may, for example be used to treat herbivores. The compositions of the present invention include geometric and optical isomers of one or more of the drugs, wherein each drug is a racemic mixture of isomers or one or more purified isomers.

Pharmaceutical compositions suitable for use in the present invention include compositions wherein the active ingredients are contained in an effective amount to achieve the intended purpose. Determination of the effective amounts is well within the capability of those skilled in the art, especially in light of the detailed disclosure provided herein.

The compounds of the present invention may exist as pharmaceutically acceptable salts. The present invention includes such salts. The term "pharmaceutically acceptable salts" is meant to include salts of active compounds which are prepared with relatively nontoxic acids or bases, depending on the particular substituent moieties found on the compounds described herein. When compounds of the present invention contain relatively acidic functionalities, base addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of the desired base, either neat or in a suitable inert solvent. Included are base addition salts such as sodium, potassium, calcium, ammonium, organic amino, or magnesium salt, or a similar salt. When compounds of the present invention contain relatively basic functionalities, acid addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of the desired acid, either neat or in a suitable inert solvent. Examples of acceptable acid addition salts include those derived from inorganic acids like hydrochloric, hydrobromic, nitric, carbonic, monohydrogencarbonic, phosphoric, monohydrogenphosphoric, dihydrogenphosphoric, sulfuric, monohydrogensulfuric, hydriodic, or phosphorous acids, as well as the salts derived from relatively nontoxic organic acids, for example, acetic, propionic, isobutyric, maleic, malonic, benzoic, succinic, suberic, fumaric, lactic, mandelic, phthalic, benzenesulfonic, p-tolylsulfonic, citric, tartaric and methanesulfonic. Also included are salts of amino acids such as arginate, and salts of organic acids like glucuronic or galactunoric (see, for example, Berge et al., "Pharmaceutical Salts", Journal of Pharmaceutical Science, 1977, 66, 1-19). Certain specific compounds of the present invention contain both basic and acidic functionalities that allow the compounds to be converted into either base or acid addition salts.

Examples of applicable salt forms include hydrochlorides, hydrobromides, sulfates, methanesulfonates, nitrates, maleates, acetates, citrates, fumarates, tartrates (eg (+)-tartrates, (-)-tartrates or mixtures thereof, including racemic mixtures), succinates, benzoates and salts with amino acids such as glutamic acid. These salts may be prepared by methods known to those skilled in art.

The neutral forms of the compounds are preferably regenerated by contacting the salt with a base or acid and isolating the parent compound in the conventional manner. The parent form of the compound differs from the various salt forms in certain physical properties, such as solubility in polar solvents.

The pharmaceutically acceptable esters in the present invention refer to non-toxic esters, preferably the alkyl esters such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl or pentyl esters, of which the methyl ester is preferred. However, other esters such as phenyl-C_1-5 alkyl may be employed if desired. Ester derivatives of certain compounds may act as prodrugs which, when absorbed into the bloodstream of a warm-blooded animal, may cleave in such a manner as to release the drug form and permit the drug to afford improved therapeutic efficacy.

Certain compounds of the present invention can exist in unsolvated forms as well as solvated forms, including hydrated forms. In general, the solvated forms are equivalent to unsolvated forms and are encompassed within the scope of the present invention. Certain compounds of the present invention may exist in multiple crystalline or amorphous forms. In general, all physical forms are equivalent for the uses contemplated by the present invention and are intended to be within the scope of the present invention.

When used as a therapeutic the compounds described herein often are administered with a physiologically acceptable carrier. A physiologically acceptable carrier is a formulation to which the compound can be added to dissolve it or otherwise facilitate its administration. Examples of physiologically acceptable carriers include, but are not limited to, water, saline, physiologically buffered saline.

Unless otherwise stated, structures depicted herein are also meant to include compounds which differ only in the presence of one or more isotopically enriched atoms. For example, compounds having the present structures except for the replacement of a hydrogen by a deuterium or tritium, or the replacement of a carbon by ¹³C- or ¹⁴C-enriched carbon are within the scope of this invention. The compounds of the present invention may also contain unnatural proportions of atomic isotopes at one or more of atoms that constitute such compounds. For example, the compounds may be radiolabeled with radioactive isotopes, such as for example tritium (3H), iodine-125 (¹²⁵I) or carbon-14 (¹⁴C). All isotopic variations of the compounds of the present invention, whether radioactive or not, are encompassed within the scope of the present invention.

A compound of the present invention can be formulated as a pharmaceutical composition. Such a pharmaceutical composition can then be administered orally, parenterally, by inhalation spray, rectally, or topically in dosage unit formulations containing conventional nontoxic pharmaceutically acceptable carriers, adjuvants, and vehicles as desired. The amount of active ingredient that can be combined with the carrier materials to produce a single dosage form varies depending upon the mammalian host treated and the particular mode of administration. Topical administration can also involve the use of transdermal administration such, as transdermal patches or iontophoresis devices. The term parenteral as used herein includes subcutaneous injections, intravenous, intramuscular, intrasternal injection, or infusion techniques. Formulation of drugs is discussed in, for example, Hoover, John E., REMINGTON'S PHARMACEUTICAL SCIENCES, Mack Publishing Co., Easton, Pa.; 1975. Other examples of drug formulations can be found in Liberman, H. A. and Lachman, L., Eds., PHARMACEUTICAL DOSAGE FORMS, Marcel Decker, New York, N.Y., 1980.

Injectable preparations, for example, sterile injectable aqueous or oleaginous suspensions can be formulated according to the known art using suitable dispersing or wetting agents and suspending agents. The sterile injectable preparation can also be a sterile injectable solution or suspension in a nontoxic parenterally acceptable diluent or solvent, for example, as a solution in 1,3-butanediol. Among the acceptable vehicles and solvents that can be employed are water, Ringer's solution, and isotonic sodium chloride solution. In addition, sterile, fixed oils are conventionally employed as a solvent or suspending medium. For this purpose any bland fixed oil can be employed including synthetic mono- or diglycerides. In addition, fatty acids such as oleic acid find use in the preparation of injectables. Dimethyl acetamide, surfactants including ionic and non-ionic detergents, polyethylene glycols can be used. Mixtures of solvents and wetting agents such as those discussed above are also useful.

Suppositories for rectal administration of the drug can be prepared by mixing the drug with a suitable nonirritating excipient such as cocoa butter, synthetic mono- di- or triglycerides, fatty acids and polyethylene glycols that are sold at ordinary temperatures but liquid at the rectal temperature and will therefore melt in the rectum and release the drug.

Solid dosage forms for oral administration can include capsules, tablets, pills, powders, and granules. In such solid dosage forms, the compounds of this invention are ordinarily combined with one or more adjuvants appropriate to the indicated route of administration. If administered per os, a compound of the invention can be admixed with lactose, sucrose, starch powder, cellulose esters of alkanoic acids, cellulose alkyl esters, talc, stearic acid, magnesium stearate, magnesium oxide, sodium and calcium salts of phosphoric and sulfuric acids, gelatin, acacia gum, sodium alginate, polyvinylpyrrolidone, and/or polyvinyl alcohol, and then tableted or encapsulated for convenient administration. Such capsules or tablets can contain a controlled-release formulation as can be provided in a dispersion of active compound in hydroxypropylmethyl cellulose. In the case of capsules, tablets, and pills, the dosage forms can also comprise buffering agents such as sodium citrate, magnesium or calcium carbonate or bicarbonate. Tablets and pills can additionally be prepared with enteric coatings.

For therapeutic purposes, formulations for parenteral administration can be in the form of aqueous or non-aqueous isotonic sterile injection solutions or suspensions. These solutions and suspensions can be prepared from sterile powders or granules having one or more of the carriers or diluents mentioned for use in the formulations for oral administration. A compound of the invention can be dissolved in water, polyethylene glycol, propylene glycol, ethanol, corn oil, cottonseed oil, peanut oil, sesame oil, benzyl alcohol, sodium chloride, and/or various buffers. Other adjuvants and modes of administration are well and widely known in the pharmaceutical art.

Liquid dosage forms for oral administration can include pharmaceutically acceptable emulsions, solutions, suspensions, syrups, and elixirs containing inert diluents commonly used in the art, such as water. Such compositions can also comprise adjuvants, such as wetting agents, emulsifying and suspending agents, and sweetening, flavoring, and perfuming agents.

The dosage regimen utilizing the compounds of the present invention in combination with an anticancer agent is selected in accordance with a variety of factors including type, species, age, weight, sex and medical condition of the patient; the severity of the condition to be treated; the route of administration; the renal and hepatic function of the patient; and the particular compound or salt or ester thereof employed. A consideration of these factors is well within the purview of the ordinarily skilled clinician for the purpose of determining the therapeutically effective dosage amounts to be given to a person in need of the instant combination therapy.

Any suitable formulation of a compound described above can be prepared for administration by methods known in the art. Selection of useful excipients or carriers can be achieved without undue experimentation, based on the desired route of administration and the physical properties of the compound to be administered.

Any suitable route of administration may be used, as determined by a treating physician, including, but not limited to, oral, parenteral, intravenous, intramuscular, transdermal, topical and subcutaneous routes. Depending on the subject to be treated, the mode of administration, and the type of treatment desired -- e.g., prevention, prophylaxis, therapy; the compounds are formulated in ways consonant with these parameters. Preparation of suitable formulations for each route of administration are known in the art. A summary of such formulation methods and techniques is found in Remington's Pharmaceutical Sciences, latest edition, Mack Publishing Co., Easton, PA. The formulation of each substance or of the combination of two substances will frequently include a diluent as well as, in some cases, adjuvants, buffers and preservatives. The substances to be administered can be administered also in liposomal compositions or as microemulsions.

For injection, formulations can be prepared in conventional forms as liquid solutions or suspensions or as solid forms suitable for solution or suspension in liquid prior to injection or as emulsions. Suitable excipients include, for example, water, saline, dextrose and glycerol. Such compositions may also contain amounts of nontoxic auxiliary substances such as wetting or emulsifying agents, and pH buffering agents, such as, for example, sodium acetate and sorbitan monolaurate, and so forth.

Various sustained release systems for drugs have also been devised, and can be applied to compounds of the invention. See, for example, U.S. patent No. 5,624,677.

Systemic administration may also include relatively noninvasive methods such as the use of suppositories, transdermal patches, transmucosal delivery and intranasal administration. Oral administration is also suitable for compounds of the invention. Suitable forms include syrups, capsules, tablets, as is understood in the art.

For administration to animal or human subjects, the appropriate dosage of a compound described above often is 0.01-15 mg/kg, and sometimes 0.1-10 mg/kg. In some embodiments, a suitable dosage of the compound of the invention for an adult patient will be between 1 and 1000 mg per dose, frequently between 10 and 300 mg, and the dosage may be administered 1-4 times per day. Dosage levels are dependent on the nature of the condition, drug efficacy, the condition of the patient, the judgment of the practitioner, and the frequency and mode of administration; optimization of such parameters is within the ordinary level of skill in the art.

Compounds of the invention may be used alone or in combination with another therapeutic agent. The invention provides methods to treat conditions such as cancer, inflammation and immune disorders by administering to a subject in need of such treatment a therapeutically effective amount of a therapeutic agent useful for treating said disorder and administering to the same subject a therapeutically effective amount of a modulator of the present invention, i.e., a compound of the invention. The therapeutic agent and the modulator may be "co-administered", i.e, administered together, either as separate pharmaceutical compositions or admixed in a single Pharmaceutical composition. By "administered together", the therapeutic agent and the modulator may also be administered separately, including at different times and with different frequencies. The modulator may be administered by any known route, such as orally, intravenously, intramuscularly, nasally, and the like; and the therapeutic agent may also be administered by any conventional route. In many embodiments, at least one and optionally both of the modulator and the therapeutic agent may be administered orally. Preferably, the modulator is an inhibitor, and it may inhibit either one of CK2 and Pim, or both of them to provide the treatment effects described herein.

In certain embodiments, a "modulator" as described above may be used in combination with a therapeutic agent that can act by binding to regions of DNA that can form certain quadruplex structures. In such embodiments, the therapeutic agents have anticancer activity on their own, but their activity is enhanced when they are used in combination with a modulator. This synergistic effect allows the therapeutic agent to be administered in a lower dosage while achieving equivalent or higher levels of at least one desired effect.

A modulator may be separately active for treating a cancer. For combination therapies described above, when used in combination with a therapeutic agent, the dosage of a modulator will frequently be two-fold to ten-fold lower than the dosage required when the modulator is used alone to treat the same condition or subject. Determination of a suitable amount of the modulator for use in combination with a therapeutic agent is readily determined by methods known in the art.

Compounds and compositions of the invention may be used in combination with anticancer or other agents, such as palliative agents, that are typically administered to a patient being treated for cancer. Such "anticancer agents" include, e.g., classic chemotherapeutic agents, as well as molecular targeted therapeutic agents, biologic therapy agents, and radiotherapeutic agents.

When a compound or composition of the invention is used in combination with an anticancer agent to another agent, the present invention provides, for example, simultaneous, staggered, or alternating treatment. Thus, the compound of the invention may be administered at the same time as an anticancer agent, in the same pharmaceutical composition; the compound of the invention may be administered at the same time as the anticancer agent, in separate pharmaceutical compositions; the compound of the invention may be administered before the anticancer agent, or the anticancer agent may be administered before the compound of the invention, for example, with a time difference of seconds, minutes, hours, days, or weeks.

In examples of a staggered treatment, a course of therapy with the compound of the invention may be administered, followed by a course of therapy with the anticancer agent, or the reverse order of treatment may be used, and more than one series of treatments with each component may also be used. In certain examples of the present invention, one component, for example, the compound of the invention or the anticancer agent, is administered to a mammal while the other component, or its derivative products, remains in the bloodstream of the mammal. For example, the present compound may be administered while the anticancer agent or its derivative products remains in the bloodstream, or the anticancer agent may be administered while the present compound or its derivatives remains in the bloodstream. In other examples, the second component, is administered after all, or most of the first component, or its derivatives, have left the bloodstream of the mammal.

The compound of the invention and the anticancer agent may be administered in the same dosage form, e.g., both administered as intravenous solutions, or they may be administered in different dosage forms, e.g., one compound may be administered topically and the other orally. A person of ordinary skill in the art would be able to discern which combinations of agents would be useful based on the particular characteristics of the drugs and the cancer involved.

Anticancer agents useful in combination with the compounds of the present invention may include agents selected from any of the classes known to those of ordinary skill in the art, including, but not limited to, antimicrotubule agents such as diterpenoids and vinca alkaloids; platinum coordination complexes; alkylating agents such as nitrogen mustards, oxazaphosphorines, alkylsulfonates, nitrosoureas, and triazenes; antibiotic agents such as anthracyclins, actinomycins and bleomycins; topoisomerase II inhibitors such as epipodophyllotoxins; antimetabolites such as purine and pyrimidine analogues and anti-folate compounds; topoisomerase I inhibitors such as camptothecins; hormones and hormonal analogues; signal transduction pathway inhibitors; nonreceptor tyrosine kinase angiogenesis inhibitors; immunotherapeutic agents; pro-apoptotic agents; and cell cycle signaling inhibitors; and other agents described below.

Anti-microtubule or anti-mitotic agents are phase specific agents that are typically active against the microtubules of tumor cells during M or the mitosis phase of the cell cycle. Examples of anti-microtubule agents include, but are not limited to, diterpenoids and vinca alkaloids.

Plant alkaloid and terpenoid derived agents include mitotic inhibitors such as the vinca alkaloids vinblastine, vincristine, vindesine, and vinorelbine; and microtubule polymer stabilizers such as the taxanes, including, but not limited to paclitaxel, docetaxel, larotaxel, ortataxel, and tesetaxel.

Diterpenoids, which are derived from natural sources, are phase specific anti - cancer agents that are believed to operate at the G2/M phases of the cell cycle. It is believed that the diterpenoids stabilize the p-tubulin subunit of the microtubules, by binding with this protein. Disassembly of the protein appears then to be inhibited with mitosis being arrested and cell death following.

Examples of diterpenoids include, but are not limited to, taxanes such as paclitaxel, docetaxel, larotaxel, ortataxel, and tesetaxel. Paclitaxel is a natural diterpene product isolated from the Pacific yew tree Taxus brevifolia and is commercially available as an injectable solution TAXOL®. Docetaxel is a semisynthetic derivative of paclitaxel q. v., prepared using a natural precursor, 10-deacetyl-baccatin III, extracted from the needle of the European Yew tree. Docetaxel is commercially available as an injectable solution as TAXOTERE®.

Vinca alkaloids are phase specific anti-neoplastic agents derived from the periwinkle plant. Vinca alkaloids that are believed to act at the M phase (mitosis) of the cell cycle by binding specifically to tubulin. Consequently, the bound tubulin molecule is unable to polymerize into microtubules. Mitosis is believed to be arrested in metaphase with cell death following. Examples of vinca alkaloids include, but are not limited to, vinblastine, vincristine, vindesine, and vinorelbine. Vinblastine, vincaleukoblastine sulfate, is commercially available as VELBAN® as an injectable solution. Vincristine, vincaleukoblastine 22-oxo-sulfate, is commercially available as ONCOVIN® as an injectable solution. Vinorelbine, is commercially available as an injectable solution of vinorelbine tartrate (NA VELBINE®), and is a semisynthetic vinca alkaloid derivative.

Platinum coordination complexes are non-phase specific anti-cancer agents, which are interactive with DNA. The platinum complexes are believed to enter tumor cells, undergo, aquation and form intra- and interstrand crosslinks with DNA causing adverse biological effects to the tumor. Platinum-based coordination complexes include, but are not limited to cisplatin, carboplatin, nedaplatin, oxaliplatin, satraplatin, and (SP-4-3)-(cis)- amminedichloro-[2-methylpyridine] platinum(II). Cisplatin, cis-diamminedichloroplatinum, is commercially available as PLATINOL® as an injectable solution. Carboplatin, platinum, diammine [1,1-cyclobutane-dicarboxylate(2-)-0,0'], is commercially available as PARAPLATIN® as an injectable solution.

Alkylating agents are generally non-phase specific agents and typically are strong electrophiles. Typically, alkylating agents form covalent linkages, by alkylation, to DNA through nucleophilic moieties of the DNA molecule such as phosphate, amino, sulfhydryl, hydroxyl, carboxyl, and imidazole groups. Such alkylation disrupts nucleic acid function leading to cell death. Examples of alkylating agents include, but are not limited to, alkyl sulfonates such as busulfan; ethyleneimine and methylmelamine derivatives such as altretamine and thiotepa; nitrogen mustards such as chlorambucil, cyclophosphamide, estramustine, ifosfamide, mechlorethamine, melphalan, and uramustine; nitrosoureas such as carmustine, lomustine, and streptozocin; triazenes and imidazotetrazines such as dacarbazine, procarbazine, temozolamide, and temozolomide. Cyclophosphamide, 2-[bis(2-chloroethyl)-amino]tetrahydro-2H-1,3,2-oxazaphosphorine 2-oxide monohydrate, is commercially available as an injectable solution or tablets as CYTOXAN®. Melphalan, 4-[bis(2-chloroethyl)amino]-L-phenylalanine, is commercially available as an injectable solution or tablets as ALKERAN®. Chlorambucil, 4-[bis(2-chloroethyl)amino]-benzenebutanoic acid, is commercially available as LEUKERAN® tablets. Busulfan, 1,4-butanediol dimethanesulfonate, is commercially available as MYLERAN® TABLETS. Carmustine, 1,3-[bis(2-chloroethyl)-1-nitrosourea, is commercially available as single vials of lyophilized material as BiCNU®, 5-(3,3-dimethyl-1-triazeno)-imidazole-4-carboxamide, is commercially available as single vials of material as DTIC-Dome®. Furthermore, alkylating agents include (a) alkylating-like platinum-based chemotherapeutic agents such as cisplatin, carboptatin, nedaplatin, oxaliplatin, satraplatin, and (SP-4-3)-(cis)-amminedichloro-[2-methylpyridine] platinum(II); (b) alkyl, sulfonates such as busulfan; (c) ethyleneimine and methylmelamine derivatives such as altretamine and thiotepa; (d) nitrogen mustards such as chlorambucil, cyclophosphamide, estramustine, ifosfamide, mechlorethamine, trofosamide, prednimustine, melphalan, and uramustine; (e) nitrosoureas such as carmustine, lomustine, fotemustine, nimustine, ranimustine and streptozocin; (f) triazenes and imidazotetrazines such as dacarbazine, procarbazine, temozolamide, and temozolomide.

Anti-tumor antibiotics are non-phase specific agents which are believed to bind or intercalate with DNA. This may result in stable DNA complexes or strand breakage, which disrupts ordinary function of the nucleic acids, leading to cell death. Examples of anti-tumor antibiotic agents include, but are not limited to, anthracyclines such as daunorubicin (including liposomal daunorubicin), doxorubicin (including liposomal doxorubicin), epirubicin, idarubicin, and valrubicin; streptomyces-related agents such as bleomycin, actinomycin, mithranrycin, mitomycin, porfiromycin; and mitoxantrone. Dactinomycin, also know as Actinomycin D, is commercially available in injectable form as COSMEGEN®. Daunorubicin, (8S-cis-)-8-acetyl-10-[(3-amino-2,3,6-trideoxy-a-L-lyxohexopyranosyl)oxy]-7,8,9,10-tetrahydro-6,8,11-trihydroxy-1-methoxy-5,12-naphthacenedione hydrochloride, is commercially available as a liposomal injectable form as DAUNOXOME® or as an injectable as CERUBIDINE®. Doxorubicin, (8S, 10S)-10-[(3-amino-2,3,6-trideoxy-α-L-lyxohexopyranosyl)oxy]-8-glycoloyl, 7,8,9,10-tetrahydro-6,8,11-trihydroxy-3-methoxy-5,12-naphthacenedione hydrochloride, is commercially available in an injectable form as RUBEX® or ADRIAMYCIN RDF®. Bleomycin, a mixture of cytotoxic glycopeptide antibiotics isolated from a strain of Streptomyces verticil/us, is commercially available as BLENOXANE®.

Topoisomerase inhibitors include topoisomerase I inhibitors such as camptothecin, topotecan, irinotecan, rubitecan, and belotecan; and topoisomerase II inhibitors such as etoposide, teniposide, and amsacrine.

Topoisomerase II inhibitors include, but are not limited to, epipodophyllotoxins, which are phase specific anti-neoplastic agents derived from the mandrake plant. Epipodophyllotoxins typically affect cells in the S and G2 phases of the cell cycle by forming a ternary complex with topoisomerase II and DNA causing DNA strand breaks. The strand breaks accumulate and cell death follows. Examples of epipodophyllotoxins include, but are not limited to, etoposide, teniposide, and amsacrine. Etoposide, 4'-demethyl-epipadophyllotoxin 9[4,6-0-(R)-ethylidene-β-D-glucopyranoside], is commercially available as an injectable solution or capsules as VePESID® and is commonly known as VP-16. Teniposide, 4'-dimethyl-epipodophyllotoxin 9[4,6-0-(R)-thenylidene-β-D-glucopyranoside], is commercially available as an injectable solution as VUMON® and is commonly known as VM-26.

Topoisomerase I inhibitors including, camptothecin and camptothecin derivatives. Examples of topoisomerase I inhibitors include, but are not limited to camptothecin, topotecan, irinotecan, rubitecan, belotecan and the various optical forms (i.e., (R), (S) or (R,S)) of 7-(4-methylpiperazino-methylene)-10, 11-ethylenedioxy-camptothecin, as described in U.S. Patent Nos. 6,063,923; 5,342,947; 5,559,235; 5,491,237 and pending U.S. patent Application No. 08/977,217 filed November 24, 1997. Irinotecan HCl, (4S)-4,11-diethyl-4-hydroxy-9-[(4-piperidinopiperidino)-carbonyloxy]-1H-yrano[3',4',6,7]indolizino[1,2-b]quinoline-3,14(4H, 12H)-dione hydrochloride, is commercially available as the injectable solution CAMPT0SAR®. Irinotecan is a derivative of camptothecin which binds, along with its active metabolite 8N-38, to the topoisomerase I-DNA complex. Topotecan HCl, (S)-10-[(dimethylamino)methyl]-4-ethyl-4,9-dihydroxy-1H-pyrano[3',4',6,7]indolizino[1,2-b]quinoline-3,14-(4H,12H)-dione monohydrochloride, is commercially available as the injectable solution HYCAMTIN®.

Anti-metabolites include (a) purine analogs such as fludarabine, cladribine, chlorodeoxyadenosine, clofarabine, mercaptopurine, pentostatin, and thioguanine; (b) pyrimidine analogs such as fluorouracil, gemcitabine, capecitabine, cytarabine, azacitidine, edatrexate, floxuridine, and troxacitabine; (c) antifolates, such as methotrexate, pemetrexed, raltitrexed, and trimetrexate. Anti-metabolites also include thymidylate synthase inhibitors, such as fluorouracil, raltitrexed, capecitabine, floxuridine and permetrexed; and ribonucleotide reductase inhibitors such as claribine, clofarabine and fludarabine. Antimetabolite neoplastic agents are phase specific anti-neoplastic agents that typically act at S phase (DNA synthesis) of the cell cycle by inhibiting DNA synthesis or by inhibiting purine or pyrimidine base synthesis and thereby limiting DNA synthesis. Consequently, S phase does not proceed and cell death follows. Anti-metabolites, include purine analogs, such as fludarabine, cladribine, chlorodeoxyadenosine, clofarabine, mercaptopurine, pentostatin, erythrohydroxynonyladenine, fludarabine phosphate and thioguanine; pyrimidine, analogs such as fluorouracil, gemcitabine, capecitabine, cytarabine, azacitidine, edatrexate, floxuridine, and troxacitabine; antifolates, such as methotrexate, pemetrexed, raltitrexed, and trimetrexate. Cytarabine, 4-amino-1-p-D-arabinofuranosyl-2 (1H)-pyrimidinone, is commercially available as CYTOSAR-U® and is commonly known as Ara-C. Mercaptopurine, 1,7-dihydro-6H-purine-6-thione monohydrate, is commercially available as PURINETHOL®. Thioguanine, 2-amino-1, 7-dihydro-6H-purine-6-thione, is commercially available as TABLOID®. Gemcitabine, 2'-deoxy-2', 2'-difluorocytidine monohydrochloride (p-isomer), is commercially available as GEMZAR®.

Hormonal therapies include (a) androgens such as fluoxymesterone and testolactone; (b) antiandrogens such as bicalutamide, cyproterone, flutamide, and nilutamide; (c) aromatase inhibitors such as aminoglutethiniide, anastrozole, exemestane, formestane, and letrozole; (d) corticosteroids such as dexamethasone and prednisone; (e) estrogens such as diethylstilbestrol; (f) antiestrogens such as fulvestrant, raloxifene, tamoxifen, and toremifine; (g) LHRH agonists and antagonists such as buserelin, goserelin, leuprolide, and triptorelin; (h) progestins such as medroxy-progesterone acetate and megestrol acetate; and (i) thyroid hormones such as levothyroxine and liothyronine. Hormones and hormonal analogues are useful compounds for treating cancers in which there is a relationship between the hormone(s) and growth and/or lack of growth of the cancer. Examples of hormones and hormonal analogues useful in cancer treatment include, but are not limited to, androgens such as fluoxymesterone and testolactone; antiandrogens such as bicalutamide, cyproterone, flutamide, and nilutamide; aromatase inhibitors such as aminogtutethimide, anastrozole, exemestane, formestane, vorazole, and letrozole; corticosteroids such as dexamethasone, prednisone and prednisolone; estrogens such as diethylstilbestrol; antiestrogens such as fulvestrant, raloxifene, tamoxifen, toremifine, droloxifene, and iodoxyfene, as well as selective estrogen receptor modulators (SERMS) such those described in U.S. Patent Nos. 5,681,835, 5,877,219, and 6,207,716; 5α-reductases such as finasteride and dutasteride; gonadotropin-releasing hormone (GnRH) and analogues thereof which stimulate the release of leutinizing hormones (LH) and/or follicle stimulating hormones (FSH), for example LHRH agonists and antagonists such as buserelin, goserelin, leuprolide, and triptorelin; progestins such as medroxyprogesterone acetate and megestrol acetate; and thyroid hormones such as levothyroxine and liothyronine.

Signal transduction pathway inhibitors are those inhibitors, which block or inhibit a chemical process which evokes an intracellular change, such as cell proliferation or differentiation. Signal tranduction inhibitors useful in the present invention include, e.g., inhibitors of receptor tyrosine kinases, non-receptor tyrosine kinases, SH2/SH3 domain blockers, serine/threonine kinases, phosphotidyl inositol-3 kinases, myo-inositol signaling, and Ras oncogenes.

Molecular targeted agents include (a) receptor tyrosine kinase ('RTK') inhibitors, such as inhibitors of EGFR, including erlotinib, gefitinib, and neratinib; inhibitors of VEGFR including vandetanib, semaxinib, and cediranib; and inhibitors of PDGFR; further included are RTK inhibitors that act at multiple receptor sites such as lapatinib, which inhibits both EGFR and HER2, as well as those inhibitors that act at each of C-kit, PDGFR and VEGFR, including but not limited to axitinib, sunitinib, sorafenib and toceranib; also included are inhibitors of BCR-ABL, c-kit and PDGFR, such as imatinib; (b) FKBP binding agents, such as an immunosuppressive macrolide antibiotic, including bafilomycin, rapamycin (surolimus) and everolimus; (c) gene therapy agents, antisense therapy agents, and gene expression modulators such as the retinoids and rexinoids, e.g. adapalene, bexarotene, trans-retinoic acid, 9-cis-retinoic acid, and N-(4-hydroxyphenyl)retinamide; (d) phenotype-directed therapy agents, including monoclonal antibodies such as alemtuzumab, bevacizumab, cetuximab, ibritumomab tiuxetan, rituximab, and trastuzumab; (e) immunotoxins such as gemtuzumab ozogamicin; (f) radioimmunoconjugates such as 131I-tositumomab; and (g) cancer vaccines.

Several protein tyrosine kinases catalyse the phosphorylation of specific tyrosyl residues in various proteins involved in the regulation of cell growth. Such protein tyrosine kinases can be broadly classified as receptor or non-receptor kinases. Receptor tyrosine kinases are transmembrane proteins having an extracellular ligand binding domain, a transmembrane domain, and a tyrosine kinase domain. Receptor tyrosine kinases are involved in the regulation of cell growth and are sometimes termed Growth factor receptors.

Inapproppriate or uncontrolled activation of many of these kinases, for example by over-expression or mutation, has been shown to result in uncontrolled cell growth. Accordingly, the aberrant activity of such kinases has been linked to malignant tissue Growth. Consequently, inhibitors of such kinases could provide cancer treatment methods.

Growth factor receptors include, for example, epidermal growth factor receptor (EGFr), platelet derived growth factor receptor (PDGFr), erbB2, erb84, vascular endothelial growth factor receptor (VEGFr), tyrosine kinase with immunoglobulin-like and epidermal growth factor homology domains (TIE-2), insulin growth factor -I (IGFI) receptor, macrophage colony stimulating factor (cfms), BTK, ckit, cmet, fibroblast growth factor (FGF) receptors, Trk receptors (TrkA, TrkB, and TrkC), ephrin (eph) receptors, and the RET protooncogene.

Several inhibitors of growth receptors are under development and include ligand antagonists, antibodies, tyrosine kinase inhibitors and anti-sense oligonucleotides. Growth factor receptors and agents that inhibit growth factor receptor function are described, for instance, in Kath, John C., Exp. Opin. Ther. Patents (2000) 10(6):803-818; Shawver et al., Drug Discov. Today (1997), 2(2):50-63; and Lofts, F. J. et al., "Growth factor receptors as targets", New Molecular Targets for Cancer Chemotherapy, ed. Workman, Paul and Kerr, David, CRC press 1994, London. Specific examples of receptor tyrosine kinase inhibitors include, but are not limited to, sunitinib, erlotinib, gefitinib, and imatinib.

Tyrosine kinases which are not growth factor receptor kinases are termed non-receptor tyrosine kinases. Non-receptor tyrosine kinases useful in the present invention, which are targets or potential targets of anti-cancer drugs, include cSrc, Lck, Fyn, Yes, Jak, cAbl, FAK (Focal adhesion kinase), Brutons tyrosine kinase, and Bcr-Abl. Such non-receptor kinases and agents which inhibit non-receptor tyrosine kinase function are described in Sinh, S. and Corey, S.J., J. Hematotherapy & Stem Cell Res. (1999) 8(5): 465 - 80; and Bolen, J.B., Brugge, J.S., Annual Review of Immunology. (1997) 15: 371-404.

SH2/SH3 domain blockers are agents that disrupt SH2 or SH3 domain binding in a variety of enzymes or adaptor proteins including, PI3-K p85 subunit, Src family kinases, adaptor molecules (Shc, Crk, Nck, Grb2) and Ras-GAP. SH2/SH3 domains as targets for anti-cancer drugs are discussed in Smithgall, T.E., J. Pharmacol. Toxicol. Methods. (1995), 34(3): 125-32. Inhibitors of Serine/Threonine Kinases including MAP kinase cascade blockers which include blockers of Raf kinases (rafk), Mitogen or Extracellular Regulated Kinase (MEKs), and Extracellular Regulated Kinases (ERKs); and Protein kinase C family member blockers including blockers of PKCs (alpha, beta, gamma, epsilon, mu, lambda, iota, zeta). IkB kinase family (IKKa, IKKb), PKB family kinases, AKT kinase family members, and TGF beta receptor kinases. Such Serine/Threonine kinases and inhibitors thereof are described in Yamamoto, T., Taya, S., Kaibuchi, K., J.Biochemistry. (1999) 126 (5): 799-803; Brodt, P, Samani, A, & Navab, R, Biochem. Pharmacol. (2000) 60:1101-1107; Massague, J., Weis-Garcia, F., Cancer Surv. (1996) 27:41-64; Philip, P.A, and Harris, AL, Cancer Treat. Res. (1995) 78: 3-27; Lackey, K. et al. Bioorg. Med. Chem. Letters, (2000) 10(3): 223-226; U.S. Patent No. 6,268,391; and Martinez-Lacaci, I., et al., Int. J. Cancer (2000),88(1): 44-52. Inhibitors of Phosphotidyl inositol-3 Kinase family members including blockers of PI3-kinase, ATM, DNA-PK, and Ku are also useful in the present invention. Such kinases are discussed in Abraham, RT. Current Opin. Immunol. (1996), 8(3): 412-8; Canman, C.E., Lim, D.S., Oncogene (1998) 17(25): 3301-8; Jackson, S.P., Int. J. Biochem. Cell Biol. (1997) 29(7):935-8; and Zhong, H. et al., Cancer Res. (2000) 60(6):1541-5. Also useful in the present invention are Myo-inositol signaling inhibitors such as phospholipase C blockers and Myoinositol analogues. Such signal inhibitors are described in Powis, G., and Kozikowski A, (1994) New Molecular Targets for Cancer Chemotherapy, ed., Paul Workman and David Kerr, CRC Press 1994, London.

Another group of signal transduction pathway inhibitors are inhibitors of Ras Oncogene. Such inhibitors include inhibitors of farnesyltransferase, geranyl-geranyl transferase, and CAAX proteases as well as anti-sense oligonucleotides, ribozymes and immunotherapy. Such inhibitors have been shown to block ras activation in cells containing wild type mutant ras , thereby acting as antiproliferation agents. Ras oncogene inhibition is discussed in Scharovsky, O.G., Rozados, V.R, Gervasoni, SI, Matar, P.,J. Biomed. Sci. (2000) 7(4): 292-8; Ashby, M.N., Curr. Opin. Lipidol. (1998) 9(2): 99 -102; and Oliff, A., Biochim. Biophys. Acta, (1999) 1423(3):C19-30.

As mentioned above, antibody antagonists to receptor kinase ligand binding may also serve as signal transduction inhibitors. This group of signal transduction pathway inhibitors includes the use of humanized antibodies to the extracellular ligand binding domain of receptor tyrosine kinases. For example Imclone C225 EGFR specific antibody (see Green, M.C. et al., Cancer Treat. Rev., (2000) 26(4): 269-286); Herceptin® erbB2 antibody (see Stern, DF, Breast Cancer Res. (2000) 2(3):176-183); and 2CB VEGFR2 specific antibody (see Brekken, R.A. et al., Cancer Res. (2000) 60(18):5117-24).

Non-receptor kinase angiogenesis inhibitors may also find use in the present invention. Inhibitors of angiogenesis related VEGFR. and TIE2 are discussed above in regard to signal transduction inhibitors (both receptors are receptor tyrosine kinases). Angiogenesis in general is linked to erbB2/EGFR signaling since inhibitors of erbB2 and EGFR have been shown to inhibit angiogenesis, primarily VEGF expression. Thus, the combination of an erbB2/EGFR inhibitor with an inhibitor of angiogenesis makes sense. Accordingly, non-receptor tyrosine kinase inhibitors may be used in combination with the EGFR/erbB2 inhibitors of the present invention. For example, anti-VEGF antibodies, which do not recognize VEGFR (the receptor tyrosine kinase), but bind to the ligand; small molecule inhibitors of integrin (alphav beta3) that will inhibit angiogenesis; endostatin and angiostatin (non-RTK) may also prove useful in combination with the disclosed erb family inhibitors. (See Bruns, CJ et al., Cancer Res. (2000), 60(11): 2926-2935; Schreiber AB, Winkler ME, & Derynck R., Science (1986) 232(4755:1250-53; Yen L. et al., Oncogene (2000) 19(31): 3460-9).

Agents used in immunotherapeutic regimens may also be useful in combination with the compounds of formula (1). There are a number of immunologic strategies to generate an immune response against erbB2 or EGFR. These strategies are generally in the realm of tumor vaccinations. The efficacy of immunologic approaches may be greatly enhanced through combined inhibition of erbB2/EGFR signaling pathways using a small molecule inhibitor. Discussion of the immunologic/tumor vaccine approach against erbB2/EGFR are found in Reilly RT, et al., Cancer Res. (2000) 60(13):3569-76; and Chen Y, et al., Cancer Res. (1998) 58(9):1965-71.

Agents used in pro-apoptotic regimens (e.g., bcl-2 antisense oligonucleotides) may also be used in the combination of the present invention. Members of the Bcl-2 family of proteins block apoptosis. Upregulation of bcl-2 has therefore been linked to chemoresistance. Studies have shown that the epidermal growth factor (EGF) stimulates anti-apoptotic members of the bcl-2 family. Therefore, strategies designed to downregulate the expression of bcl-2 in tumors have demonstrated clinical benefit and are now in Phase II/III trials, namely Genta's G3139 bcl-2 antisense oligonucleotide. Such pro-apoptotic strategies using the antisense oligonucleotide strategy for bcl-2 are discussed in Waters JS, et al., J. Clin. Oncol. (2000) 18(9): 1812-23; and Kitada S, et al. Antisense Res. Dev. (1994) 4(2): 71-9.

Cell cycle signalling inhibitors inhibit molecules involved in the control of the cell cycle. A family of protein kinases called cyclin dependent kinases (CDKs) and their interaction with a family of proteins termed cyclins controls progression through the eukaryotic cell cycle. The coordinate activation and inactivation of different cyclin/CDK complexes is necessary for normal progression through the cell cycle. Several inhibitors of cell cycle signaling are under development. For instance, examples of cyclin dependent kinases, including CDK2, CDK4, and CDK6 and inhibitors for the same are described in, for instance, RosaniaGR & Chang Y-T., Exp. Opin. Ther. Patents (2000) 10(2):215-30.

Other molecular targeted agents include FKBP binding agents, such as the immunosuppressive macrolide antibiotic, rapamycin; gene therapy agents, antisense therapy agents, and gene expression modulators such as the retinoids and rexinoids, e.g. adapalene, bexarotene, traus-retinoic acid, 9-cisretinoic acid, and N-(4 hydroxyphenyl)retinamide; phenotype-directed therapy agents, including: monoclonal antibodies such as alemtuzumab, bevacizumab, cetuximab, ibritumomab tiuxetan, rituximab, and trastuzumab; immunotoxins such as gemtuzumab ozogamicin, radioimmunoconjugates such as 131-tositumomab; and cancer vaccines.

Anti-tumor antibiotics include (a) anthracyclines such as daunorubicin (including liposomal daunorubicin), doxombicin (including liposomal doxorubicin), epirubicin, idarubicin, and valrubicin; (b) streptomyces-related agents such as bleomycin, actinomycin, mithramycin, mitomycin, portiromycin; and (c) anthracenediones, such as mitoxantrone and pixantrone. Anthracyclines have three mechanisms of action: intercalating between base pairs of the DNA/RNA strand; inhibiting topoisomerase II enzyme; and creating iron-mediated free oxygen radicals that damage the DNA and cell membranes. Anthracyclines are generally characterized as topoisomerase II inhibitors.

Monoclonal antibodies include, but are not limited to, murine, chimeric, or partial or fully humanized monoclonal antibodies. Such therapeutic antibodies include, but are not limited to antibodies directed to tumor or cancer antigens either on the cell surface or inside the cell. Such therapeutic antibodies also include, but are not limited to antibodies directed to targets or pathways directly or indirectly associated with CK2. Therapeutic antibodies may further include, but are not limited to antibodies directed to targets or pathways that directly interact with targets or pathways associated with the compounds of the present invention. In one variation, therapeutic antibodies include, but are not limited to anticancer agents such as Abagovomab, Adecatumumab, Afutuzumab, Alacizumab pegol, Alemtuzumab, Altumomab pentetate, Anatumomab mafenatox, Apolizumab, Bavituximab, Belimumab, Bevacizumab, Bivatuzumab mertansine, Blinatumomab, Brentuximab vedotin, Cantuzumab mertansine, Catumaxomab, Cetuximab, Citatuzumab bogatox, Cixutumumab, Clivatuzumab tetraxetan, Conatumumab, Dacetuzumab, Detumomab, Ecromeximab, Edrecolomab, Elotuzumab, Epratuzumab, Ertumaxomab, Etaracizumab, Farletuzumab, Figitumumab, Fresolimumab, Galiximab, Glembatumumab vedotin, Ibritumomab tiuxetan, Intetumumab, Inotuzumab oxogamicin, Ipilimumab, Iratumumab, Labetuzumab, Lexatumumab, Lintuzumab, Lucatumumab, Lumiliximab, Mapatumumab, Matuzumab, Milatuzumab, Mitumomab, Nacolomab tafenatox, Naptumomab estafenatox, Necitumumab, Nimotuzumab, Ofatumumab, Olaratumab, Oportuzumab monatox, Oregovomab, Panitumumab, Pemtumomab, Pertuzumab, Pintumomab, Pritumumab, Ramucirumab, Rilotumumab, Rituximab, Robatumumab, Sibrotuzumab, Tacatuzumab tetraxetan, Taplitumomab paptox, Tenatumomab, Ticilimumab, Tigatuzumab, Tositumomab, Trastuzumab, Tremelimumab, Tucotuzumab celmoleukin, Veltuzumab, Volociximab, Votumumab, Zalutumumab, and Zanolimumab. In some embodiments, such therapeutic antibodies include, alemtuzumab, bevacizumab, cetuximab, daclizumab, gemtuzumab, ibritumomab tiuxetan, pantitumumab, rituximab, tositumomab, and trastuzumab; in other embodiments, such monoclonal antibodies include alemtuzumab, bevacizumab, cetuximab, ibritumomab tiuxetan, rituximab, and trastuzumab; alternately, such antibodies include daclizumab, gemtuzumab, and pantitumumab. In yet another embodiment, therapeutic antibodies useful in the treatment of infections include but are not limited to Afelimomab, Efungumab, Exbivirumab, Felvizumab, Foravirumab, Ibalizumab, Libivirumab, Motavizumab, Nebacumab, Pagibaximab, Palivizumab, Panobacumab, Rafivirumab, Raxibacumab, Regavirumab, Sevirumab, Tefibazumab, Tuvirumab, and Urtoxazumab. In a further embodiment, therapeutic antibodies can be useful in the treatment of inflammation and/or autoimmune disorders, including, but are not limited to, Adalimumab, Atlizumab, Atorolimumab, Aselizumab, Bapineuzumab, Basiliximab, Benralizumab, Bertilimumab, Besilesomab, Briakinumab, Canakinumab, Cedelizumab, Certolizumab pegol, Clenoliximab, Daclizumab, Denosumab, Eculizumab, Edobacomab, Efalizumab, Erlizumab, Fezakinumab, Fontolizumab, Fresolimumab, Gantenemmab, Gavilimomab, Golimumab, Gomiliximab, Infliximab, Inolimomab, Keliximab, Lebrikizumab, Lerdelimumab, Mepolizumab, Metelimumab, Muromonab-CD3, Natalizumab, Ocrelizumab, Odulimomab, Omalizumab, Otelixizumab, Pascolizumab, Priliximab, Reslizumab, Rituximab, Rontalizumab, Rovelizumab, Ruplizumab, Sifalimumab, Siplizumab, Solanezumab, Stamulumab, Talizumab, Tanezumab, Teplizumab, Tocilizumab, Toralizumab, Ustekinumab, Vedolizumab, Vepaliznoznab, Visilizumab, Zanolimumab, and Zolimomab aritox. In yet another embodiment, such therapeutic antibodies include, but are not limited to adalimumab, basiliximab, certolizumab pegol, eculizumab, efalizurnab, infliximab, muromonab-CD3, natalizumab, and omalizumab. Alternately the therapeutic antibody can include abciximab or ranibizumab. Generally a therapeutic antibody is non-conjugated, or is conjugated with a radionuclide, cytokine, toxin, drug-activating enzyme or a drug-filled liposome.

Akt inhibitors include 1L6-Hydroxymethyl-chiro-inositol-2-(R)-2-O-methyl-3-O-octadecyl-sn-glycerocarbonate, SH-5 (Calbiochem Cat. No. 124008), SH-6 (Calbiochem Cat. No. Cat. No. 124009), Calbiochem Cat. No. 124011, Triciribine (NSC 154020, Calbiochem Cat. No. 124012), 10-(4'-(N-diethylamino)butyl)-2-chlorophenoxazine, Cu(II)Cl₂(3-Formylchromone thiosemicarbazone), 1,3-dihyrdro-1-(1-((4-(6-phenyl-1H-imidazo[4,5-g]quinoxalin-7-yl)phenyl)methyl)-4-piperidinyl)-2H-benzimidazol-2-one, GSK690693 (4-(2-(4-amino-1,2,5-oxadiazol-3-yl)-1-ethyl-7-{[(3S)-3-piperidinylmethyl]oxy}-1H-imidazo[4,5-c]pyridin-4-yl)-2-methyl-3-butyn-2-ol), SR13668 ((2,10-dicarbethoxy-6-methoxy-5,7-dihydro-indolo[2,3-b] carbazole), GSK2141795, Perifosine, GSK21110183, XL418, XL147, PF-04691502, BEZ-235 [2-Methyl-2-[4-(3-methyl-2-oxo-8-quinolin-3-yl-2,3-dihydro-imidazo[4,5-c]quinolin-1-yl)-phenyl]-propionitrile], PX-866 ((acetic acid (1S,4E,10R,11R,13S,14R)-[4-diallylaminomethylene-6-hydroxy-1-methoxymethyl-10,13-dimethyl-3,7,17-trioxo-1,3,4,7,10,11,12,13,14,15,16,17-dodecahydro-2-oxa-cyclopenta[a]phenanthren-11-yl ester)), D-106669, CAL-101, EDC0941 (2-(1H-indazol-4-yl)-6-(4-methanesulfonyl-piperazin-1-ylmethyl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidine), SF1126, SF1188, SF2523, TG100-115 [3-[2,4-diamino-6-(3-hydroxyphenyl)pteridin-7-yl]phenol]. A number of these inhibitors, such as, for example, BEZ-235, PX-866, D 106669, CAL-101, GDC0941, SF1126, SF2523 are also identified in the art as PI3K/mTOR inhibitors; additional examples, such as PI-103 [3-[4-(4-morpholinylpyrido[3',2':4,5]furo[3,2-d]pyrimidin-2-yl]phenol hydrochloride] are well-known to those of skill in the art. Additional well-known PI3K inhibitors include LY294002 [2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one] and wortmannin. mTOR inhibitors known to those of skill in the art include temsirolimus, deforolimus, sirolimus, everolimus, zotarolimus, and biolimus A9. A representative subset of such inhibitors includes temsirolimus, deforolimus, zotarolimus, and biolimus A9.

HDAC inhibitors include (i) hydroxamic acids such as Trichostatin A, vorinostat (suberoylanilide hydroxamic acid (SAHA)), panobinostat (LBH589) and belinostat (PXD101) (ii) cyclic peptides, such as trapoxin B, and depsipeptides, such as romidepsin (NSC 630176), (iii) benzamides, such as MS-275 (3-pyridylmethyl-N-{4-[(2-aminophenyl)-carbamoyl]-benzyl}-carbamate), CI994 (4-acetylamino-N-(2aminophenyl)-benzamide) and MGCD0103 (N-(2-aminophenyl)-4-((4-(pyridin-3-yl)pyrimidin-2-ylamino)methyl)benzamide), (iv) electrophilic ketones, (v) the aliphatic acid compounds such as phenylbutyrate and valproic acid.

Hsp90 inhibitors include benzoquinone ansamycins such as geldanamycin, 17-DMAG (17-Dimethylamino-ethylamino-17-demethoxygeldanamycin), tanespimycin (17-AAG, 17-allylamino-17-demethoxygeldanamycin), EC5, retaspimycin (IPI-504, 18,21-didehydro-17-demethoxy-18,21-dideoxo-18,21-dihydroxy-17-(2-propenylamino)-geldanamycin), and herbimycin; pyrazoles such as CCT 018159 (4-[4-(2,3-dihydro-1,4-benzodioxin-6-yl)-5-methyl-1H-pyrazol-3-yl]-6-ethyl-1,3-benzenediol); macrolides, such as radicocol; as well as BHB021 (CNF2024), SNX-5422, STA-9090, and AUY922.

Miscellaneous agents include altretamine, arsenic trioxide, gallium nitrate, hydroxyurea, levamisole, mitotane, octreotide, procarbazine, suramin, thalidomide, lenalidomide, photodynamic compounds such as methoxsalen and sodium porfimer, and proteasome inhibitors such as bortezomib.

Biologic therapy agents include: interferons such as interferon-α2a and interferon-α2b, and interleukins such as aldesleukin, denileukin diftitox, and oprelvekin.

In addition to these anticancer agents intended to act against cancer cells, combination therapies including the use of protective or adjunctive agents, including: cytoprotective agents such as armifostine, dexrazonxane, and mesna, phosphonates such as parmidronate and zoledronic acid, and stimulating factors such as epoetin, darbepoetin, filgrastim, PEG-filgrastim, and sargramostim, are also envisioned.

The following examples illustrate the invention. Compounds outside the scope of the present claims are included as reference embodiments.

To the reaction flask, 5,7-dichloropyrazolo[1,5-a]pyrimidine (3 g, 16 mmol) was added along with ammonium hydroxide solution (48 mL). The heterogeneous reaction was refluxed at 85°C for 12 hours. After cooling to room temperature, the mixture was filtered, washed with water, and dried under vacuum overnight. The product, 5-chloropyrazolo[1,5-a]pyrimidin-7-amine, was collected as an off-white solid in 88% yield. LCMS (M+1=169)

To the reaction flask, 5-chloropyrazolo[1,5-a]pyrimidin-7-amine (2.4 g, 14.1 mmol) was added to dichloromethane (35 mL) along with di-tert-butyl dicarbonate (3.7 g, 17 mmol), triethylamine (2.4 mL, 17 mmol) and DMAP (100 mg, 0.8 mmol). The reaction was stirred at room temperature for 6 hours then diluted with DCM, washed with saturated NaHCO₃ solution (3x) followed by washing with brine. The organic layer was isolated, dried over anhydrous MgSO₄, filtered, and evaporated to dryness. The product, tert-butyl 5-chloropyrazolo[1,5-a]pyrimidin-7-ylcarbamate, was collected as an off-white solid in 98% yield. LCMS (M-t-Butyl =213)

To tert-butyl 5-chloropyrazolo[1,5-a]pyrimidin-7-ylcarbamate (3.7 g, 13.8 mmol)) in DMF (36 mL), POCl₃ (7.7 mL, 82.9 mmol) was added dropwise at 0°C. After the addition was complete, the reaction was allowed to warm to room temperature and stirred for 8 hours. Then, the reaction was quenched by slow addition to ice cold 6N NaOH. The mixture was diluted with water then the solid was collected by filtration. The solid was washed several more times with water and dried under vacuum overnight. The product, tert-butyl 5-chloro-3-formylpyrazolo[1,5-a]pyrimidin-7-ylcarbamate, was collected as a solid in 27% yield. The product did not ionize on LCMS unless first deprotected using TFA/DCM (1:1). LCMS (M+1=197)

Tert-butyl 5-chloro-3-formylpyrazolo[1,5-a]pyrimidin-7-ylcarbamate (1.1 g, 3.8 mmol) was added to 1,4-dioxane (15 mL) along with 3-chloroaniline (2.4 mL, 22.6 mmol) and p-toluenes ulfonic acid monohydrate (73 mg, 0.4 mmol). The reaction was heated at 95°C for 12 hours then cooled to room temperature, diluted with water and filtered. Analysis of the recovered solid by LCMS showed product mass (M+1 = 288), as well as, product with chloro aniline imine mass (M+1 = 397). To completely convert this mixture to the desired product, the solid was dissolved in 6 mL of MeOH/conc. HCl solution (1:1) and heated at 60°C for 1.5 hours. The reaction was quenched by slow addition to ice cold 6N NaOH. The mixture was diluted with water then the solid was collected by filtration. The solid was washed several more times with water then dried under vacuum overnight. The product, 7-amino-5-(3-chlorophenylamino)pyrazolo[1,5-a]pyrimidine-3-carbaldehyde, was collected as orange-red solid in 38% yield. LCMS(M+1=288)

To the reaction vial, 7-amino-5-(3-chlorophenylamino)pyrazolo[1,5-a]pyrimidine-3-carbaldehyde (411 mg, 1.4 mmol) was added to ethanol (5.2 mL) along with Hydantoin (143 mg, 1.4 mmol) and piperidine (141 µL, 1.4 mmol). The reaction was heated at 80°C for 60 minutes in the microwave. The reaction was then cooled to room temperature and diluted with water. The solid was collected by filtration, washed with water and cold ethanol. The material was dried under vacuum overnight. The product, 5-((7-amino-5-(3-chlorophenylamino)pyrazolo[1,5-a]pyrimidin-3-yl)methylene)imidazolidine-2,4-dione, was recovered as a red solid in 54% yield. LCMS (M+1 = 370)

To the reaction vial, 5-((7-amino-5-(3-chlorophenylamino)pyrazolo[1,5-a]pyrimidin-3-yl)methylene)imidazolidine-2,4-dione (15 mg, 0.04 mmol) was added to DMF (0.2 mL) along with HBTU (30 mg, 0.08 mmol), DIEA (28 µL, 0.16 mmol) and 1-(tert-butoxycarbonyl)piperidine-4-carboxylic acid (18 mg, 0.08 mmol). The reaction was stirred at room temperature for 8 hours then heated at 95°C for 4 hours. The reaction was then cooled to room temperature and diluted with water. The solid was collected by filtration, washed with water, 1N HCl solution, and more water. The material was then dissolved in 5% DCM/MeOH and purified by prep HPLC. The isolated fractions were combined and evaporated to dryness. The material was dissolved in 1 mL of TFA/DCM (1:1) and stirred at room temperature for 1 hour. The solvent was removed by evaporation under a stream of nitrogen and the crude material was washed with 1N NaOH followed by water. The solid was collected by filtration and dried under vacuum overnight. The product, N-(5-(3-chlorophenylamino)-3-((2,5-dioxoimidazolidin-4-ylidene)methyl)pyrazolo[1,5-a]pyrimidin-7-yl)piperidine-4-carboxamide, was recovered as a solid in 2% yield. LCMS (M+1 = 481).

Table 1 below shows the biological acvities of Examples 5 and 6 as listed as Compounds A1 and B1. Compound CK2: IC50 (µM) PIM2: IC50 (5 µM ATP) AB: MDAMB453 (µM) AB: BxPC3 (µM) A1 <0.1 1.1779 2.464 16.599 B1 <1.0 2.5000

To 5,7-dichloropyrazolo[1.5-a]pyrimidine (200 mg, 1.06 mmol) in ACN was added Et₃N (148 µl, 1.06 mmol) and cyclopropylamine (75µl, 1.06 mmol). The reaction was refluxed at 80°C overnight. The mixture was concentrated under reduced pressure, dissolved in DCM, and washed with water. The resulting organic layer was dried over Na₂SO₄ and concentrated under reduced pressure to afford 156 mg of 5-chloro-N-cyclopropylpyrazolo[1,5-a]pyrimidin-7-amine (70% yield). LCMS (M+1=209)

To 5-chloro-N-cyclopropylpyrazolo[1,5-a]pyrimidin-7-amine (156 mg, 0.75 mmol) in DMF was added POCl₃ (205 µl, 2.25 mmol). The mixture was stirred at room temperature for 3 hours. Ice was added to quench excess POCl₃ then the mixture was neutralized with 1M NaOH. DCM was added and the product was extracted three times. The organic layer was dried over Na₂SO₄ and concentrated under reduced pressure to yield 5-chloro-7-(cyclopropylamino)pyrazolo[1,5-a]pyrimidine-3-carbaldehyde. Some residual DMF could not be removed. LCMS (M+1=237)

To 5-chloro-7-(cyclopropylamino)pyrazolo[1,5-a]pyrimidine-3-carbaldehyde (177 mg, 0.75 mmol) in 1,4-dioxane was added 3-chloroaniline (397 µl, 3.75 mmol). The mixture was heated in microwave at 120°C for 60 minutes. The precipitate was filtered off, and the filtrate was purified by prep TLC (1%MeOH/DCM) to yield 26 mg (11% yield) of 5-(3-chlorophenylamino)-7-(cyclopropylamino)pyrazolo[1,5-a]pyrimidine-3-carbaldehyde. LCMS (M+1=328)

To 5-(3-chlorophenylamino)-7-(cyclopropylamino)pyrazolo[1,5-a]pyrimidine-3-carbaldehyde (26 mg, 0.08 mmol) in EtOH was added hydantoin (8 mg, 0.08 mmol) and piperidine (8 µl, 0.08 mmol). The mixture was stirred at 70°C over the weekend. Insolubilities were filtered off, and filtrate was concentrated under reduced pressure. Filtrate was then dissolved in MeOH and isolated by prep HPLC to yield 5-((5-(3-chlorophenylamino)-7-(cyclopropylamino)pyrazolo[1,5-a]pyrimidin-3-yl)methylene)imidazolidine-2,4-dione. LCMS (M+1=410)

Tert-butyl 5-chloro-3-formylpyrazolo[1,5-a]pyrimidin-7-yl(cyclopropyl)carbamate (0.2 g, 0.59 mmol) was suspended in ethanol (2 mL). 3-fluoroaniline (189 mg, 1.48 mmol) was added, followed by 4M HCl/dioxane (0.3 mL, 1.18 mmol). The reaction was heated to 80 °C for 6 h, and then the volatiles were removed in vacuo. The residue was diluted with water (10 mL) and the pH was adjusted to 12 by the addition of 6M NaOH. The solution was stirred for 0.5 h, then the precipitate, which is a mixture of tert-butyl cyclopropyl(5-(3-fluorophenylamino)-3-formylpyrazolo[1,5-a]pyrimidin-7-yl)carbamate and the corresponding imine, were isolated by filtration and dried in vacuo. The imine was hydrolyzed by dissolving in methanol (9 mL), 1,4-dioxane (3.6 mL) and 6M HCl (9 mL) and heating at 60 °C for 5 h. The solution was poured onto ice (50 mL) and the pH was adjusted to 12 by addition of 6M NaOH. The precipitate was isolated by filtration and dried in vacuo to provide tert-butyl cyclopropyl(5-(3-fluorophenylamino)-3-formylpyrazolo[1,5-a]pirimidin-7-yl)carbamate (172 mg, 93%). LCMS (M+1=312)

Hydantoin (69 mg, 0.69 mmol) and piperidine (69 µL, 0.69 mmol) were added to 7-(cyclopropylamino)-5-(3-fluorophenylamino)pyrazolo[1,5-a]pyrimidine-3-carbaldehyde (72 mg, 0.23 mmol) dissolved in ethanol (1.1 mL). The reaction was heated at 80 °C. After 15 h, the reaction was cooled to r.t., diluted with water (5 mL), and the precipitate was collected and washed with 1:1 ethanol:water (5 mL). The bright yellow solid was dried in vacuo to give (Z)-5-((7-(cyclopropylamino)-5-(3-fluorophenylamino)pyrazolo[1,5-a]pyrimidin-3-yl)methylene)imidazolidine-2,4-dione (25 mg, 10% over 3 steps). LCMS (M+1=507)

The compounds described in the following Table 2A were prepared using chemistries similar to those exemplified in Example 11 and Example 12. All compounds were characterized by LCMS. Table 2B shows the biological activities of the compounds listed in Table 2A. Compound CK2: IC50 (µM) PIM2: IC50 (µM) PIM2: IC50 (5 µM ATP) AB: MDAMB453 (µM) AB: BxPC3 (µM) C1 <0.01 1.3 >2.5000 1.038 15.029 D1 <0.01 2.2002 1.34 4.067 E1 <0.01 >2.5000 17.713 16.835 F1 <0.01 >2.5000 2.037 5.763

3,5-Difluoroaniline (29 mg, 0.22 mmol), Cs₂CO₃ (67 mg, 0.21 mmol) were added to Tert-butyl 5-chloro-3-formylpyrazolo[1,5-a]pyrimidin-7-yl(cyclopropyl)carbamate (50 mg, 0.15 mmol) dissolved in 1,4-dioxane (1 mL). Racemic BINAP (6 mg, 0.06 mmol) and palladium(II) acetate (4 mg, 0.04 mmol) were then added. The mixture was sealed and irradiated at 110 °C for 20 min in the microwave. Et₂O (3 mL) was added and the solution was filtered. The filtrate was concentrated in vacuo. The crude residue was dissolved in dichloromethane (1.5 mL) and trifluoroacetic acid (1.5 mL). After 1 h, the solution was concentrated under a stream of air. The residue was triturated with 20% 2-propanol/hexanes. The product was filtered to yield 7-(cyclopropylamino)-5-(3,5-difluorophenylamino)pyrazolo[1,5-a]pyrimidine-3-carbaldehyde (39 mg, 80%). LCMS (M+1=330)

The compounds described in the following Table 3 were prepared using chemistries similar to those exemplified in Example 13. All compounds were characterized by LCMS. Structure MW LCMS m/z [M+1]+ Structure MW LCMS m/z [M+1]+ 395.7 396 345.8 346 294.3 295 345.8 346 327.7 328 351:4 352 357.7 358 368.1 369 361.3 362 294.3 295 345.8 346 347.4 348 359.4 360 267.3 268

Hydantoin (28 mg, 0.28 mmol) and piperidine (42 µL, 0.42 mmol) were added to 7-(cyclopropylamino)-5-(3,5-difluorophenylamino)pyrazolo[1,5-a]pyrimidine-3-carbaldehyde (52 mg, 0.16 mmol) dissolved in ethanol (1 mL). The reaction was heated at 80°C. After 12 h, the reaction was cooled to r.t., diluted with water (2 mL), and the precipitate was collected and washed with 1:1 ethanol:water (5 mL). The solid was dried in vacuo to give (Z)-5-((7-(cyclopropylamino)-5-(3,5-difluorophenylamino)pyrazolo[1,5-a]pyrimidin-3-yl)methylene)imidazolidine-2,4-dione (18 mg, 28% over 3 steps). LCMS (M+1=440)

The compounds described in the following tables were prepared using chemistries similar to those exemplified in Example 14. All compounds were characterized by LCMS. Table 4B shows the biological activities of the compounds listed in Table 4A. Compound CK2: IC50 (µM) PIM2: IC50 (5 µM ATP) AB: MDAMB453 (µM) AB: BxPC3 (µM) G1 <0.01 > 2.5000 10.006 >30 H1 <0.01 > 2.5000 0.991 3.209 I1 <0.01 > 2.5000 11.121 15.148 J1 <0.01 2.4799 7.116 3.924 K1 <0.01 > 2.5000 7.711 5.66 L1 <0.01 > 2.5000 0.5 1.354 M1 <0.01 > 2.5000 N1 <.0.01 1.9706 O1 <0.01 > 2.5000 P1 <0.01 > 2.5000 Q1 <0.01 > 2.5000 R1 <0.01 > 2.5000 S1 <0.01 2.041

The compounds described in the following Table 5 were prepared following the general scheme below using chemistries similar to Example 13 and Example 14.

To tert-butyl 5-chloro-3-formylpyrazolo[1, 5-a]pyrimidin-7-yl (cyclopropylmethyl) carbamate (50 mg, 0.14 mmol) in 1 mL of 1,4 dioxane was added cesium carbonate (65 mg, 0.2 mmol), Pd(OAc)₂ (4 mg, 0.006 mmol), (±)-BINAP (5 mg, 0.009 mmol), 3-chloro-4-fluoroaniline (31 mg, 0.21 mmol). The reaction mixture was heated in microwave at 110°C for 20 minutes. The mixture was then cooled to room temperature, water was added, and the product was extracted with ether. The organic layer was then concentrated under reduced pressure and the crude product was dissolved in 1:1 mixture of dichloromethane and trifluoroacetic acid at room temperature for 1hour. The reaction mixture was concentrated with 10 mL of dichloromethane. To the reaction mixture, ether/ hexanes (1:1) was added and the flask was sonicated for 10 minutes then filtered to obtain the yellow precipitate. The precipitate was washed with hexane to yield 5-(3-chloro-4-fluorophenylamino)-7-(cyclopropylmethylamino) pyrazolo [1, 5-a] pyrimidine-3-carbaldehyde. LCMS (M+1=460)

To 5-(3-chloro-4-fluorophenylamino)-7-(cyclopropylmethylamino) pyrazolo [1, 5-a] pyrimidine-3-carbaldehyde (40 m.g, 0.09 mmol) in 1.0 mL of ethanol was added hydantoin (9 mg, 0.09 mmol) and piperdine (8 ul). The reaction was heated at 80°C overnight, cooled to room temperature, filtered, and washed with ethanol to yield 20 mg (31% yield) (Z)-5-((5-(3-chloro-4-fluorophenylamino)-7-(cyclopropylmethylamino) pyrazolo [1, 5-a] pyrimidin-3-yl) methylene) imidazolidine-2,4-dione as a yellow powder. LCMS (M+1=442)

The compounds described in the following tables were prepared using chemistries similar to those exemplified in Example 15 and Example 16. All compounds were characterized by LCMS. Table 6B shows the biological activities of the compounds listed in Table 6A. Compound CK2: IC50 (µM) PIM2: IC50 (5 µM ATP) AB: MDAMB453 (µM) AB: BxPC3 (µM) T1 <0.01 0.5087 2.196 12.315 U1 <0.1 > 2.5000 V1 <0.1 > 2.5000 5.086 6.188 W1 <0.1 > 2.5000 7.424 9.207 X1 <0.1 > 2.5000 6.935 7.986 Y1 <0.1 > 2.5000 Z1 <0.1 A2 <0.1 B2 <0.1 C2 <0.1 D2 <0.1 E2 <0.01 F2 <0.1 G2 <0.01 H2 <0.01 I2 <0.1

To tert-butyl 5-chloro-3-formylpyrazolo [1,5-a] pyrimidin-7-yl (cyclopropyl) carbamate (500 mg 1.5 mmol) in dimethylformamide was added sodium azide (150 mg, 2.3 mmol) then the reaction mixture was stirred at room temperature for 30 minutes. The reaction mixture was then partitioned between ethyl acetate/water. The organic layer was collected, dried over sodium sulfate, filtered, and concentrated under high vacuum to yield tert-butyl 5-azido-3-formylpyrazolo[1,5-a]pyrimidin-7-yl(cyclopropyl)carbamate. The crude product was taken to next step without further purification. LCMS (M+1=344)

The crude product, tert-butyl 5-azido-3-formylpyrazolo [1, 5-a] pyrimidin-7-yl (cyclopropyl) carbamate was subjected to hydrogenation using 10% wt palladium on carbon in ethanol. The reaction was stirred under hydrogen for 3 hours. The mixture was filtered through celite and sonicated with 1:1 mixture of ethyl acetate and hexane. The yellow solid was filtered and dried under reduced pressure and was dissolved in (1:1) DCM/TFA at room temperature for 1 hour. The reaction mixture was washed with aqueous sodium bicarbonate solution and extracted with dichloromethane. The organic layer was concentrated and dried under high vacuum to yield 5-amino-7-(cyclopropylamino) pyrazolo [1, 5-a] pyrimidine-3-carbaldehyde as product 310 mg (95% yield on three steps). LCMS (M+1=218)

To 5-amino-7-(cyclopropylamino) pyrazolo [1, 5-a] pyrimidine-3-carbaldehyde (75 mg, 0.34 mmol) in 1.0 mL ethanol was added hydantoin (34 mg, 0.34 mmol) and piperdine (33 ul). The reaction was heated at 80°C overnight. The reaction mixture was cooled to room temperature and yellow precipitate was filtered, washed with ethanol to yield 45 mg (44% yield) Z)-5-((5-amino-7-(cyclopropylamino)pyrazolo[1,5-a]pyrimidin-3-yl)methylene)imidazolidine-2,4-dione. LCMS (M+1=300)

To 5-amino-7-(cyclopropylamino)pyrazolo[1,5-a]pyrimidine-3-carbaldehyde (50 mg, 0.23 mmol) in 1.0 mL tetrahydrofuran was added methyl chloroformate (35 ul, 0.46 mmol) and DIEA (39 ul). The reaction mixture was heated at 60°C for one hour. The reaction was partitioned between ethyl acetate/water. The organic layer was col lected, dried over sodium sulfate, and concentrated under high vacuum to yield methyl 7-(cyclopropylamino)-3-formylpyrazolo[1,5-a]pyrimidin-5-ylcarbamate. The crude product was taken to next step without further purification. LCMS (M+1=276)

To methyl 7-(cyclopropylamino)-3-formylpyrazolo [1, 5-a] pyrimidin-5-ylcarbamate (33 mg, 0.12 mmol) in 1.0 mL ethanol was added Hydantoin (12 mg, 0.12 mmol) and piperdine (11 ul). The reaction was heated at 80°C for two hours. The reaction was cooled to room temperature, the yellow precipitate was filtered, and washed with ethanol to yield 15 mg (40% yield) (Z)-methyl 7-(cyclopropylamino)-3-((2, 5-dioxoimidazolidin-4-ylidene) methyl) pyrazolo [1, 5-a] pyrimidin-5-ylcarbamate. LCMS (M+1=358)

To 5-amino-7-(cyclopropylamino) pyrazolo [1, 5-a] pyrimidine-3-carbaldehyde (step b) (58 mg, 0.266 mmol) in 1.0 mL tetrahydrofuran was added cyclopropane carbonyl chloride (38 ul, 0.419mmol) and DIPEA (39.0 ul). The reaction mixture was heated at 60°C for one hour. The reaction was partitioned between ethyl acetate and water, the organic layer was dried under sodium sulfate concentrated on high vac to yield N-(7-(cyclopropylamino)-3-formylpyrazolo[1,5-a]pyrimidin-5-yl) cyclopropanecarboxamide. The crude product was taken to next step without further purification. LCMS (M+1=286)

To N-(7-(cyclopropylamino)-3-formylpyrazolo [1, 5-a] pyrimidin-5-yl) cyclopropanecarboxamide (74 mg, 0.26 mmol) in 1.0 mL ethanol was added hydantoin (26 mg, 0.26 mmol) and piperdine (24 ul). The reaction was heated at 80°C for two hours. The reaction mixture was cooled to room temperature, concentrated and diluted with MeOH. The product was purified by prep HPLC to yield 14 mg (20% yield) (Z)-N-(7-(cyclopropylamino)-3-((2,5-dioxoimidazolidin-4-ylidene)methyl)pyrazolo[1,5-a]pyrimidin-5-yl)cyclopropanecarboxamide. LCMS (M+1=368)

The compounds described in the following table were prepared using chemistries similar to those exemplified in Example 20 and Example 16. All compounds were characterized by LCMS. Table 7B shows the biological activities of the compounds listed in Table 7A. Compound CK2: IC50 (µM) PIM2: IC50 (5 µM ATP) AB: MDAMB453 (µM) AB: BxPC3 (µM) J2 <0.1 > 2.5000 K2 <0.01 > 2.5000 11.768 9.93 L2 <0.01 4.17 10.986

Tert-butyl 5-chloro-3-formylpyrazolo[1,5-a]pyrimidin-7-yl(cyclopropyl)carbarmate (0.5 g, 1.48 mmol) was dissolved in glacial acetic acid (5 mL). NaOAc (1.21g, 14.8 mmol) and Hydantoin (356 mg, 3.56 mmol) were added and the reaction was placed in a 110 °C bath for 4 d. The solution was cooled to r.t. and water (15 mL) was added. The precipitate was filtered and then triturated with ethanol (5 mL) and then CH₂Cl₂ (5 mL) to give (Z)-5-((5-(chloro-7-(cyclopropylamino)pyrazolo[1,5-a]pyrimidin-3-yl)methylene)imidazolidine-2,4-dione (202 mg, 43%). LCMS (ES): >85% pure, m/z 319 [M+1]⁺.

(Z)-5-((5-(Chloro-7-(cyclopropylamino)pyrazolo[1,5-a]pyrimidin-3-yl)melhylene)imidazolidine-2,4-dione (25 mg, 0.08 mmol) was suspended in NMP (0.2 mL). 4,4-difluoropiperidine hydrochloride (60 mg, 0.38 mmol) and diisopropylethylamine (67 µL, 0.38 mmol) were added and the reaction was irradiated in the microwave at 140°C for 20 min. The product was purified by preparative HPLC to afford (Z)-5-((7-(cyclopropylamino)-5-(4,4-difluoropiperidin-1-yl)pyrazolo[1,5-a]pyrimidin-3-yl)methylene)imidazolidine-2,4-dione (5.9 mg, 14%). LCMS (ES): >95% pure, m/z 404 [M+1]⁺.

Tert-butyl 5-chloro-3-formylpyrazolo[1,5-a]pyrimidin-7-yl(cyclopropyl)carbamate (1.0 eq, 105 mg, 0.312 mmol) was dissolved in DMF (1 ml) in a vial. K₂CO₃ (1.5 eq, 64 mg, 0.463 mmol) and cyclopentylamine (1.5 eq, 46 ul, 0.465 mmol) were added and the mixture was stirred at 50°C for one hour. An additional amount of cyclopentylamine (1.5 eq, 46 ul, 0.465 mmol) was added and the mixture stirred at 70°C for 2 hours. Water was added and the resulting precipitate was filtered and dried to provide 130 mg of solid. This solid was stirred in HCl 4N in dioxane (4 ml) at room temperature for 4 hours. Methanol (1 ml) and aqueous 6N HCl (2 ml) were added and the mixture stirred at room temperature overnight. The reaction was subsequently stirred overnight at 60°C to complete the imine hydrolysis. The reaction was neutralized with 6N NaOH and the compound extracted with methylene chloride. After drying over Na₂SO₄, and evaporation of-the volatiles, the material was triturated in ethylacetate to form a solid. 5-(cyclopentylamino)-7-(cyclopropylamino)pyrazolo[1,5-a]pyrimidine-3-carbaldehyde was isolated as a solid by filtration (21mg). LCMS (ES):>95% pure, m/z 286 [M+H]⁺.

5-(Cyclopentylamino)-7-(cyclopropylamino)pyrazolo[1,5-a]pyrimidine-3-carbaldehyde (1.0 eq, 20 mg, 0.070 mmol) was mixed in a vial with hydantoin (2.8 eq, 20 mg, 0.20 mmol) in Ethanol (0.3 ml). Piperidine (2.9 eq, 20 ul, 0.202 mmol) was added and the mixture was stirred at 90°C for 3 hours. The mixture was cooled down, the precipitate was filtered, washed with ethanol and dried. (Z)-5-((5-(cyclopentylamino)-7-(cyclopropylamino)pyrazolo[1,5-a]pyrimidin-3-yl)methylene)imidazolidine-2,4-dione was isolated as a solid (25 mg, 100%). LCMS (ES):>95% pure, m/z 368 [M+H]⁺.

The compounds listed in the following Table 6 are Example 25, Example 26, and Example 27. Compound CK2: IC50 (µM) PIM2: IC50 (5 µM ATP) AB: MDAMB453 (µM) AB: BxPC3 (µM) M2 <0.01. 0.359 6.676 4.872 N2 <0.01 0.8453 3.042 9.185 O2 <0.01 > 2.5000 4.514 10.417

(R,Z)-5-((7-(Cyclopropylamino)-5-(3-fluoropyrrolidin-1-yl)pyrazolo[1,5-a]pyrimidin-3-yl)methylene)imidazolidine-2,4-dione was prepared using chemistries similar to the ones used to prepare (Z)-5-((5-(cyclopentylamino)-7-(cyclopropylamino)pyrazolo[1,5-a]pyrimidin-3-yl)methylene)imidazolidine-2,4-dione. The compound was isolated as a solid (84 mg, 75% yield). LCMS (ES):>95% pure, m/z 372 [M+H]⁺.

5-Chloro-3-formylpyrazolo[1,5-a]pyrimidin-7-yl(cyclopropyl)carbamate (1.0 eq, 113 mg, 0.335 mmol) was mixed in a vial with trans-tert-butyl 4-aminocyclohexylcarbamate (1.0 eq, 81 mg, 0.335 mmol) and K₂CO₃ (5.0 eq, 232 mg, 1.68 mmol) in DMF (1 ml). The mixture was stirred at 70°C for 2.5 hours. Water was added, the solid was filtered and dried. The compound was treated with hydantoin (3.0 eq, 100 mg), piperidine (3.0 eq, 100 ul) in ethanol (2 ml) at 85-90°C for 4.5 hours. Water was added and the solid was filtered and dried. The crude solid was suspended in methylene chloride (5 ml) and trifluoroacetic acid (1 ml) and the mixture was stirred at room temperature for 1 hour. The volatiles were evaporated. The residue was dissolved in methanol and water and subjected to purification by preparative HPLC. After genevac evaporation (Z)-5-((5-((1r,4r)-4-aminocyclohexylamino)-7-(cyclopropylamino)pyrazolo[1,5-a]pyrimidin-3-yl)methylene)imidazolidine-2^.4-dione 2,2,2-trifluoroacetate was isolated a yellow solid (96 mg, 56% yield). LCMS (ES):>99% pure, m/z 397 [M+H]⁺. Two isomers detected (ratio: 97.5% and 2.5%).

The following compounds were prepared using chemistries similar to the one used in Example 26, Example 27, Example 28 and Example 29. Compounds were characterized by LCMS. Table 9B shows the biological activities of the compounds listed in Table 9A. Compounds CK2: IC50 (µM) PIM2: IC50 (5 µM ATP) AB: MDAMB453 (µM) AB: BxPC3 (µM) P2 <0.01 0.7076 0.66 2.12 Q2 <0.01 0.2841 0.154 5.371 R2 <0.1 0.8783 S2 <0.01 1.2984 3.507 3.876 T2 <0.01 0.9978 2.494 6.901 U2 <0.01 0.7659 12.773 > 30 V2 <0.01 1.3652 1.483 1.626 W2 <0.01 0.8771 X2 <0.01 1.952

(Z)-5-((5-((1r,4r)-4-Aminocyclohexylamino)-7-(cyclopropylamino)pyrazolo[1,5-a]pyrimidin-3-yl)methylene)imidazolidine-2,4-dione 2,2,2-trifluoroacetate (1.0 eq, 10 mg, 0.0196 mmol) and DIEA (1.2 eq, 4 ul, 0.0229 mmol) were dissolved in NMP (0.1 ml). Acetic anhydride (1.0 eq, 2 ul, 0.0211 mmol) was added and the mixture stirred at room temperature overnight. Water was added and the resulting precipitate was filtered and dried to provide N-((1r,4r)-4-(7-(cyclopropylamino)-3-((Z)-(2,5-dioxoimidazolidin-4-ylidene)methyl)pyrazolo[1,5-a]pyrimidin-5-ylamino)cyclohexyl)acetamide as a solid (8 mg). LCMS (ES):>95% pure, m/z 439 [M+H]⁺.

(Z)-5-((5-((1r,4r)-4-Aminocyclohexylamino)-7-(cyclopropylamino)pyrazolo[1,5-a]pyrimidin-3-yl)methylene)imidazolidine-2,4-dione 2,2,2-trifluoroacetate (10 mg) and DIEA (1.2 eq, 4.1 ul) were mixed in dry NMP (0.1 ml). Dimethylcarbamic chloride (1.0 eq, 1.8 ul) was added and the mixture stirred at room temperature overnight. The reaction was diluted with NMP (1.5 ml) and a few drops of water. The compound was purified by preparative HPLC and was isolated after evaporation at the genevac. 3-((1r,4r)-4-(7-(cyclopropylamino)-3-((Z)-(2,5-dioxoimidazolidin-4-ylidene)methyl)pyrazolo[1,5-a]pyrimidin-5-ylamino)cyclohexyl)-1,1-dimethylurea 2,2,2-trifluoroacetate. LCMS (ES):>95% pure, m/z 468 [M+H]⁺. Z:E ratio: 86:13

The following molecules were prepared using similar chemistries to the ones described in Example 30 and Example 31 using the appropriate amines and anhydrides or acyl chlorides, sulfamoyl chlorides, sulfonyl chlorides or chloroformates. Compounds were purified by preparative HPLC, isolated after genevac evaporation, and characterized by LCMS. Table 10B shows the biological activities of the compounds listed in Table 10A. Compound CK2: IC50 (µM) PIM2: IC50 (5 µM ATP) AB: MDAMB453 (µM) AB: BxPC3 (µM) A3 <0.01 >2.5000 >30 >30 B3 <0.1 0.5385 C3 <0.01 0.6791 D3 <0.01 0.476 E3 <0.01 >2.5000

The following molecules in Table 10 were prepared using chemistries similar to the ones in Example 26, Example 27, Example 28 and Example 29.

Diisopropylethylamine (256 µL, 1.48 mmol) was added to tert-butyl 5-chloro-3-formylpyrazolo[1,5-a]pyrimidin-7-yl(cyclopropyl)carbamate (250 mg, 0.74 mmol) suspended in ethanol (2.5 mL). Benzyl mercaptan (191 µL, 1.48 mmol) was added and the reaction was homogeneous after ∼2 min. After 10 min, the reaction was diluted with ethanol (3 mL) and the precipitate was filtered and washed with ethanol (10 mL) to yield tert-butyl 5-(benzylthio)-3-formylpyrazolo[1,5-a]pyrimidin-7-yl(cyclopropyl)carbamate (148 mg, 47%). LCMS (ES): >95% pure, m/z 425 [M+1]⁺.

Hydantoin (67 mg, 0.67 mmol) and piperidine (66 µL, 0.67 mmol) were added to tert-butyl 5-(benzylthio)-3-formylpyrazolo[1,5-a]pyrimidin-7-yl(cyclopropyl)carbamate (95 mg, 0.22 mmol) dissolved in ethanol (1.1 mL). The reaction was heated at 80 °C. After 15 h, the reaction was cooled to r.t., diluted with water (5 mL), and the precipitate was collected and washed with 1:1 ethanol:water (10 mL) and then ethanol (3 mL). The bright yellow solid was dried in vacuo to give (Z)-5-((5-(benzylthio)-7-(cyclopropylamino)pyrazolo[1,5-a]pyrimidin-3-yl)methylene)imidazolidine-2,4-dione (75 mg, 66%). LCMS (ES): >95% pure, m/z 507 [M+1]⁺.

5-((5-(Benzylthio)-7-(cyclopropylamino)pyrazolo[1,5-a]pyrimidin-3-yl)methylene)imidazolidine-2,4-dione (60 mg, 0.15 mmol) was dissolved in dichloromethane (1 mL) and trifluoroacetic acid (1 mL). After 1 h, the solution was concentrated under a stream of air. The residue was triturated with Et₂O (3 mL) and the precipitate was collected to provide (Z)-5-((5-(benzylthio)-7-(cyclopropylamino)pyrazolo[1,5-a]pyrimidin-3-yl)methylene)imidazolidine-2,4-dione (59 mg, 98%). LCMS (ES): >95% pure, m/z 407 [M+1]⁺ Structure LCMS m/z [M+1]+ CK2: IC50 (uM) PIM2: IC50 (5um ATP) AB: MDAMB453 (uM) AB: BxPC3 (uM) 407 <0.01 > 2.5000 > 30 11.037

To 5-chloro-7-(cyclopropylamino)pyrazolo[1,5-a]pyrimidine-3-carbaldehyde (440 mg, 1.86 mmol) in EtOH was added thiazolidine-2,4-dione (458 mg, 3.91 mmol) and piperidine (208 µl, 2.05 mmol). The reaction was heated at 80°C overnight. To the reaction mixture, isopropanol (3 mL) was added in the along with 218 mg thiazolidine-2,4-dione and 94µL of piperidine. The temperature was increased to 90°C and the reaction was stirred at that temperature overnight. The precipitate was filtered while hot and dissolved in MeOH. To the reaction mixture, 1M HCl (1 mL) was added and the mixture sonicated. The precipitate was filtered and washed with MeOH to yield 340 mg (54% yield) 5-((5-chloro-7-(cyclopropylamino)pyrazolo[1,5-a]pyrimidin-3-yl)methylene)thiazolidine-2,4-dione as a yellow powder. LCMS (M+1=336)

To 5-((5-chloro-7-(cyclopropylamino)pyrazolo[1,5-a]pyrimidin-3-yl)methylene)thiazolidine-2,4-dione (20 mg, 0.06 mmol) in NMP was added 3-chloroaniline (38 µL, 0.36 mmol) and a few granules of p-toluenesulfonic acid. The reaction was heated in microwave at 180°C 1.5 hours. The reaction mixture was filtered and purified by prep HPLC then prep TLC (1%MeOH/DCM) to yield 5-((5-(3-chlorophenylamino)-7-(cyclopropylamino)pyrazolo[1,5-a]pyrimidin-3-yl)methylene)thiazolidine-2,4-dione as a yellow solid. LCMS (M+1=427)

To 5-((5-chloro-7-(cyclopropylamino)pyrazolo[1,5-a]pyrimidin-3-yl)methylene)thiazolidine-2,4-dione (30 mg, 0.09 mmol) in NMP was added 2-methylpropan-1-amine (20 mg, 0.268 mmol). The reaction was heated at 130°C overnight. The reaction mixture was diluted with MeOH and purified by prep HPLC to yield 5-((7-(cyclopropylamino)-5-(isobutylamino)pyrazolo[1,5-a]pyrimidin-3-yl)methylene)thiazolidine-2,4-dione. LCMS (M+1=373)

The compounds described in the following table were prepared using chemistries similar to those exemplified in Example 36. All compounds were characterized by LCMS. Table 12B shows the biological activities of the compounds listed in Table 12A. Compound CK2: IC50 (µM) PIM1: IC50 (30 µM ATP) PIM2: IC50 (5 µM ATP) AB: MDAMB453 (µM) AB: BxPC3 (µM) F3 <1.0 2.0638 1.6438 G3 <0.1 2.0575 1.8456 7.7 9.45 H3 <1.0 2.3875 > 2.5000 I3 <0.1 > 2.5000 0.8759 J3 <1.0 > 2.5000 > 2.5000 K3 <1.0 0.9177 1.3934 L3 <1.0 > 2.5000 1.4327 M3 <0.1 1.4455 1.4379 N3 <1.0 > 2.5000 > 2.5000 O3 <0.1 1.2533 > 2.5000 P3 <1.0 > 2.5000 > 2.5000 Q3 <0.1 2.0461 R3 <1.0 1.82

To 5-chloro-7-(cyclopropylamino)pyrazolo[1,5-a]pyrimidine-3-carbaldehyde (4.52 g, 19.15 mmol) in methylene chloride (80 mL) was added triethylamine (3.2 mL, 23 mmol), dimethylaminopyridine (350 mg, 2.87 mmol), and di-t-butyldicarbonate (12.53 g, 57.44 mmol) The mixture was stirred at room temperature for 60 minutes. The reaction mixture was transferred to a separatory funnel, washed 1X with H₂O, 2X with brine, dried over MgSO₄, filtered, and evaporated to dryness to provide an oily residue. The crude material was purified by silica gel chromatography (0%-20% ethyl acetate/hexanes) to yield 5.68 g (88% yield) of tert-butyl 5-chloro-3-formylpyrazolo[1,5-a]pyrimidin-7-yl (cyclopropyl)carbamate. LCMS (M+1=337)

To 5 tert-butyl 5-chloro-3-formylpyrazolo[1,5-a]pyrimidin-7-yl (cyclopropyl)carbamate (650 mg, 1.93 mmol) in 14 mL of a 2:1 mixture of 1,2-dimethoxyethane/ EtOH was added 3-hydroxyphenyl boronic acid (399 mg, 2.89 mmol), tetrakis(triphenylphosphine)palladium(0) (112 mg, 0.096 mmol), and 2M aqueous solution of Na₂CO₃ (2.9 mL, 5.79 mmol). The mixture was stirred at 85°C for 1h. The volatiles were removed by rotary evaporation and the residue was purified by silica gel chromatography (0%-30% EtOAc/Hexanes) to provide 400 mg of tert-butyl cyclopropyl(3-formyl-5-(3-hydroxyphenyl)pyrazolo[1,5-a]pyrimidin-7-yl)carbamate. (52%). LCMS (M+1=395)

To tert-butyl cyclopropyl(3-formyl-5-(3-hydroxyphenyl)pyrazolo[1,5-a]pyrimidin-7-yl)carbamate (400 mg, 1.01 mmol) in methylene chloride (20 mL) was added TFA (10 mL). The reaction mixture was stirred at room temperature for 2 hours. The volatiles were removed by rotary evaporation and the residue was purified by silica gel chromatography (0%-40% EtOAc/hexanes) to provide 103 mg of 7-(cyclopropylamino)-5-(3hydroxyphenyl)pyrazolo [1,5-a]pyrimidine-3-carbaldehyde. (35%). LCMS (M+1=295)

To 7-(cyclopropylamino)-5-(3hydroxyphenyl)pyrazolo [1,5-a]pyrimidine-3-carbaldehyde (100 mg, 0.340 mmol) in EtOH (2 mL) was added piperidine (67 µL, 0.680 mmol), and hydantoin (34 mg, 0.34 mmol). The reaction was stirred at 50°C overnight. The solid formed was isolated by filtration to provide 70 mg of 5-((7-(cyclopropylamino)-5-(3-hydroxyphenyl)pyrazolo[1,5-a]pyrimidin-3-yl)methylene)imidazolidine-2,4-dione. (55%). LCMS (M+1=377)

The compounds described in the following table were prepared using chemistries similar to those exemplified in Example 38, Example 39, and Example 40. All compounds were characterized by LCMS. Table 13B shows the biological activities of the compounds listed in Table 13A. Compound CK2: IC50 (µM) PIM2: IC50 (5 µM ATP) AB: MDAMB453 (µM) AB: BxPC3 (µM) S3 <0.01 0.6135 0.33 10.627 T3 <0.01 0.8908 0.402 1.541 U3 <0.01 > 2.5000 3.743 3.68 V3 <0.01 0.6492 0.477 5.171 W3 <0.01 > 2.5000 1.709 2.054 X3 <0.01 > 2.5000 0.517 13.111 Y3 <0.01 > 2.5000 Z3 <0.01 2.2144 0.284 2.916 A4 <0.01 > 2.5000 > 30 > 30 B4 <0.01 2.1685 7.866 7.907 C4 <0.1 2.4032 1.494 3.279 D4 <0.01 > 2.5000 1.054 23.617 E4 <0.01 > 2.5000 1.174 11.78 F4 <0.01 > 2.5000 1.298 6.592 G4 <0.01 > 2.5000 1.153 1.191 H4 <0.01 > 2.5000 1.964 14.486 I4 <0.1 > 2.5000 0.683 1.898 J4 <0.01 0.867 4.746 >30 K4 <0.1 1.3082 1.938 2.578 L4 <0.0.1 1.4748 1.79 0.725 M4 <0.01 1.2497 >30 14.437 N4 <0.01 > 2.5000 > 30 12.535 O4 <0.01 > 2.5000 17.123 1.232 P4 <0.01 0.0754 5.276 0.549 Q4 <0.01 0.2562 1.068 0.745 R4 <0.01 0.0487 14.882 10.61 S4 <0.01 > 2.5000 20.012 4.608 T4 <0.1 > 2.5000 1.706 2.744 U4 <0.01 > 2.5000 1.263 8.129 V4 <0.1 > 2.5000 12.417 > 30 W4 <0.01 2.084 12.278 > 30 X4 <0.01 1.7271 > 30 > 30 Y4 <0.01 > 2.5000 1.979 2.253 Z4 <0.01 > 2.5000 15.69 29.035 A5 <0.01 0.948 1.742 B5 <0.01 > 2.5000 26.74 5.426 C5 <1.0 > 2.5000

Same procedure as [Example 38]. LCMS (M+1=441)

Same procedure as [Example 39]. LCMS (M+1=341)

To 5-(7-(cyclopropylamino)-3-formylpyrazolo[1,5-a]pyrimidin-5-yl)-2-fluorobenzoic acid (75 mg, 0.22 mmol 0 in DMF (3 mL) was added EDCI (46 mg, 0.24 mmol), HOBt (33 mg, 0.24 mmol), and morpholine (21 mg, 0.24 mmol). The reaction mixture was stirred at room temperature for 2 hours. The reaction mixture was diluted with ethyl acetate and washed 1X with saturated sodium bicarbonate, 2X with brine, dried over MgSO₄, filtered and evaporated to dryness to provide 92 mg of 7-(cyclopropylamino)-5-(4-fluoro-3-(morpholine-4-carbonyl)phenyl)pyrazolo[1,5-a]pyrimidine-3-carbaldehyde. LCMS (M+1=410)

Same procedure as [Example 40]. LCMS (M+1=492)

The compounds described in the following table were prepared using chemistries similar to those exemplified in Example 42 and Example 43. All compounds were characterized by LCMS. Table 14B shows the biological activities of the compounds listed in Table 14A. Compound CK2: IC50 (µM) PIM2: IC50 (5 µM ATP) AB: MDAMB453 (µM) AB: BxPC3 (µM) D5 <0.1 > 2.5000 2.443 1.208 E5 <0.1 > 2.5000 F5 <0.01 > 2.5000 0.948 1.808 G5 <0.01 > 2.5000 0.435 1.841

To tert-butyl 5-chloro-3-formylpyrazolo[1,5-a]pyrimidin-7-yl(cyclopropyl)carbamate (1 g, 3 mmol) in 29 niL of a 2:1 mixture of 1,2-dimethoxyethane/ EtOH was added 2-Fluoropyridine-4-boronic acid (500 mg, 3.55 mmol), tetrakis(triphenylphosphine)palladium(0) (173 mg, 0.15 mmol), and 2M aqueous solution of Na₂CO₃ (4.4 mL, 8.9 mmol). The mixture was stirred at 85°C for 8 hours. The volatiles were removed by rotary evaporation and the residue was purified by silica gel chromatography (35% EtOAc/Hexanes) to provide 324 mg tert-butyl cyclopropyl(5-(2-fluoropyridin-4-yl)-3-formylpyrazolo[1,5-a]pyrimidin-7-yl)carbamate (28% yield). LCMS (M+1=398)

To tert-butyl cyclopropyl(5-(2-fluoropyridin-4-yl)-3-formylpyrazolo[1,5-a]pyrimidin-7-yl)carbamate (320 mg, 0.82 mmol) in methylene chloride (3 mL) was added TFA (3 mL). The reaction mixture was stirred at room temperature for 1.5 hours. The volatiles were removed by rotary evaporation and 1N NaOH was added to the residue to make basic. The precipitate was collected by filtration, washed with water, and dried under vacuum to provide 180 mg of 7-(cyclopropylamino)-5-(2-fluoropyridin-4-yl)pyrazolo[1,5-a]pyrimidine-3-carbaldehyde (74%). LCMS (M+1=298)

To 7-(cyclopropylamino)-5-(2-fluoropyridin-4-yl)pyrazolo[1,5-a]pyrimidine-3-carbaldehyde 30 mg, 0.1 mmol) in EtOH (1 mL) was added piperdine (13 µL, 0.1 mmol), and thiazolidine-2,4-dione (12 mg, 0.1 mmol). The reaction was stirred at 80°C for 2 hours. The solid formed was isolated by filtration, washed with water then ethanol. The recovered solid was further purified by washing with 20% methanol/dichloromethan to provide 9 mg of 5-((7-(cyclopropylamino)-5-(2-fluoropyridin-4-yl)pyraxolo[1,5-a]pyrimidin-3-yl)methylene)thiazolidine-2,4-dione (23%). LCMS (M+1=397)

The compounds described in the following table were prepared using chemistries similar to those exemplified in Example 44, Example 45, and Example 46. All compounds were characterized by LCMS. Table 15B shows the biological activities of the compounds listed in Table 15A. Compound CK2: IC50 (µM) PIM2: IC50 (5 µM ATP) AB: MDAMB453 (µM) AB: BxPC3 (µM) H5 <0.1 0.8152 I5 <1.0 0.2176 J5 <1.0 0.1978 > 30 18.951

To 7-(cyclopropylamino)-5-(2-fluoropyridin-4-yl)pyrazolo[1,5-a]pyrimidine-3-carbaldehyde 30 mg, 0.1 mmol) in EtOH (1 mL) was added piperdine (13 µL, 0.1 mmol), and rhodanine (13 mg, 0.1 mmol). The reaction was stirred at 80°C for 2 hours. The solid formed was isolated by filtration, washed with water then ethanol. The recovered solid was further purified by washing with 20% methanol/dichloromethan to provide 15 mg of 5-((7-(cyclopropylamino)-5-(2-fluoropyridin-4-yl)pyrazolo[1,5-a]pyrimidin-3-yl)methylene)-2-thioxothiazolidin-4-one (35%). LCMS (M+1=413) Structure LCMS m/z [M+1]+ CK2: IC50 (uM) PIM2: IC50 (5um ATP) AB: MDAMB453 (uM) AB: BxPC3 (uM) 413 <1.0 1.5908 > 30 22.671

To the reaction flask, 5,7-dichloropyrazolo[1,5-a]pyrimidine (896 mg, 4.8 mmol) was added along with tert-butyl 4-aminopiperidine-1-carboxylate (954 mg, 4.8 mmol), triethylamine (664 µL, 4.8 mmol), and acetonitrile(16 mL). The reaction was heated at 100°C for 12 hours then cooled to room temperature, diluted with water, filtered and washed with water. The product, tert-butyl 4-(5-chloropyrazolo[1,5-a]pyrimidin-7-ylamino)piperidine-1-carboxylate, was collected as a solid in quantitative yield and dried under vacuum overnight. LCMS (M+1=352)

To tert-butyl 4-(5-chloropyrazolo[1,5-a]pyrimidin-7-ylamino)piperidine-1-carboxylate (1.7 g, 4.8 mmol) in DMF (36 mL), POCl₃ (7.7 mL, 82.9 mmol) was added dropwise at room temperature. After the addition was complete, the reaction was stirred for 8 hours. Then, the reaction was quenched by slow addition to ice cold 6N NaOH. The mixture was diluted with water and the solid was collected by filtration. The solid was washed several more times with water then dried under vacuum overnight. The product, tert-butyl 4-(5-chloro-3-formylpyrazolo[1,5-a]pyrimidin-7-ylamino)piperidine-1-carboxylate, was collected as a solid in 48% yield. LCMS (M+1=380)

Tert-butyl 4-(5-chloro-3-formylpyrazolo[1,5-a]pyrimidin-7-ylamino)piperidine-1-carboxylate (876 mg, 2.3 mmol) was added to 1,4-dioxane (6 mL) along with 3-chloroaniline (1.5 mL, 13.9 mmol) and p-toluenesulfonic acid monohydrate (44 mg, 0.23 mmol). The reaction was heated at 95°C for 12 hours then cooled to room temperature, diluted with water, and filtered. The solid was washed with 1N NaOH followed with water then dried under vacuum overnight. The product, tert-butyl 4-(5-(3-chlorophenylamino)-3-formylpyrazolo[1,5-a]pyrimidin-7-ylamino)piperidine-1-carboxylate, was collected after further purification by recrystallization from ethyl acetate/hexanes (74% yield). LCMS (M+1=471)

To the reaction flask, tert-butyl 4-(5-(3-chlorophenylamino)-3-formylpyrazolo[1,5-a]pyrimidin-7-ylamino)piperidine-1-carboxylate (811 mg, 1.7 mmol) was added to ethanol (6.3 mL) along with hydantoin (172 mg, 1.7 mmol) and piperidine (170 µL, 1.7 mmol). The reaction was heated at 80°C for 12 hours then cooled to room temperature and diluted with water. The solid was collected by filtration, washed with water and cold ethanol. The material was dried under vacuum overnight. The product, tert-butyl 4-(5-(3-chlorophenylamino)-3-((2,5-dioxoimidazolidin-4-ylidene)methyl)pyrazolo[1,5-a]pyrimidin-7-ylamino)piperidine-1-carboxylate, was recovered as a red solid in 67% yield after further purification by recrystallization from ethyl acetate/hexanes. LCMS (M+1= 553)

Tert-butyl 4-(5-(3-chlorophenylamino)-3-((2,5-dioxoimidazolidin-4-ylidene)methyl)pyrazolo[1,5-a]pyrimidin-7-ylamino)piperidine-1-carboxylate (640 mg, 1.2 mmol) was dissolved in 10 mL of TFA/DCM (1:1) and stirred at room temperature for 1 hour then quenched by addition to ice cold 6N NaOH. The mixture was diluted with water then the aqueous layer was decanted. The organic layer was diluted with hexanes and filtered. The product, 5-((5-(3-chlorophenylamino)-7-(piperidin-4-ylamino)pyrazolo[1,5-a]pyrimidin-3-yl)methylene)imidazolidine-2,4-dione, was collected as a solid in quantitative yield. LCMS (M+1 = 453)

To 5-((5-(3-chlorophenylamino)-7-(piperidin-4-ylamino) pyrazolo [1, 5-a] pyrimidin-3-yl) methylene) imidazolidine-2,4-dione (30 mg, 0.066 mmol) in THF was added cyclopropyl carbonyl chloride (5 µL, 0.04 mmol). The mixture was stirred at room temperature for ten minutes. The reaction mixture was then concentrated, diluted with MeOH, and purified by prep HPLC to yield 5-((5-(3-chlorophenylamino)-7-(1-(cyclopropanecarbonyl)piperidin-4-ylamino)pyrazolo[1,5-a]pyrimidin-3-yl)methylene)imidazolidine-2,4-dione. LCMS (M+1=521)

Same procedure as [Example 53]. LCMS(M+1=537)

Same procedure as [Example 53]. LCMS (M+1=551)

Same procedure as [Example 53]. LCMS (M+1=524).

Same procedure as [Example 53] except DME is used as solvent. LCMS (M+1=511)

Same procedure as [Example 53] except DMF is used as solvent. LCMS (M+1=525)

To 5-((5-(3-chlorophenylamino)-7-(piperidin-4-ylamino)pyrazolo[1,5-a]pyrimidin-3-yl)methylene)imidazolidine-2,4-dione (30 mg, 0.066 mmol) in DMF was added 1-chloro2-propanol (7 µL, 0.13 mmol) and potassium iodide (11.0 mg,0.066 mmol). The mixture was heated to 120°C and stirred for overnight. The reaction mixture was concentrated , diluted with MeOH, and purified by prep HPLC to yield 5-((5-(3-chlorophenylamino)-7-(1-(2-hydroxypropyl)piperidin-4-ylamino)pyrazolo[1,5-a]pyrimidin-3-yl)methylene)imidazolidine-2,4-done. LCMS (M+1=511)

Same procedure as [Example 59]. LCMS (M+1=497)

To 5-((5-(3-chlorophenylamino)-7-(piperidin-4-ylamino)pyrazolo [1,5-a] pyrimidin-3-yl) methylene) imidazolidine-2,4-dione (30 mg, 0.066 mmol) in DMF was added 2-(bromomethyl) pyridine hydrogen bromide (26.0 mg, 0103 mmol). The mixture was stirred at room temperature for 0.5 hour. The reaction mixture was concentrated, diluted with MeOH, and purified by prep HPLC to yield 5-((5-(3-chlorophenylamino)-7-(1-(pyridin-2-ylmethyl)piperidin-4-ylamino)pyrazolo[1,5-a]pyrimidin-3-yl)methylene;)imidazolidine-2,4-dione. LCMS (M+1=544)

To 5-((5-(3-chlorophenylamino)-7-(piperidin-4-ylamino) pyrazolo [1, 5-a] pyrimidin-3-yl) methylene) imidazolidine-2,4-dione (20 mg, 0.04 mmol) in THF and AcOH (4.8 mg, 0.08 mmol) was added acetone (2.0 mL, 0.2 mmol) and sodium triacetoxy borohydride (85.0 mg, 0.4 mmol)). The mixture was heated at 60°C for one hour. Saturated sodium bicarbonate solution was added to the reaction mixture. The mixture was extracted with ethyl acetate and dried over sodium sulfate. Then the mixture was , diluted with MeOH, and purified by prep HPLC to yield 5-((5-(3-chlorophenylamino)-7-(1-isopropylpiperidin-4-ylammo) pyrazolo [1, 5-a] pyrimidin-3-yl) methylene) imidazolidine-2, 4-dione. LCMS (M+1=495)

To 5-((5-(3-chlorophenylamino)-7-(piperidin-4-ylamino) pyrazolo [1, 5-a] pyrimidin-3-yl) methylene) imidazolidine-2,4-dione (30 mg, 0.06 mmol) in THF and AcOH (4.8 mg, 0.08 mmol) was added acetaldehyde (2.0 mL, 0.2 mmol) and sodium triacetoxy borohydride (85.0 mg, 0.4 mmol). The mixture was stirred at room temperature for 0.5 hour. The reaction mixture was concentrated , diluted with MeOH, and purified by prep HPLC to yield 5-((5-(3-chlorophenylamino)-7-(1-ethylpiperidin-4-ylamino)pyrazolo[1,5-a] pyrimidin-3-yl) methylene) imidazolidine-2, 4-dione. LCMS (M+1=481)

To 5-((5-(3-chlorophenylamino)-7-(piperidin-4-ylamino)pyrazolo [1, 5-a] pyrimidin-3-yl) methylene) imidazolidine-2,4-dione (30 mg, 0.06 mmol) in THF and AcOH (4.8 mg, 0.08 mmol) was added isobutryldehyde (2.2 mL, 0.2 mmol) and sodium triacetoxy borohydride (85.0 mg, 0.4 mmol). The mixture was stirred at room temperature for 0.5 hour. The mixture was concentrated, diluted with MeOH, and purified by prep HPLC to yield 5-((5-(3-chlorophenylamino)-7-(1-isobutylpiperidin-4-ylamino) pyrazolo [1, 5-a] pyrimidin-3-yl) methylene) imidazolidine-2, 4-dione. LCMS (M+1=509)

The compounds described in the following table were prepared using chemistries similar to those exemplified in the Examples described above. All compounds were characterized by LCMS. Table 17B shows the biological activities of the compounds listed in Table 17A. Compound CK2: IC50 (µM) PIM2: IC50 (5 µM ATP) AB: MDAMB453 (µM) AB: BxPC3 (µM) K5 <0.1 2.4491 1.078 10.984 L5 <0.01 > 2.5000 0.993 2.399 M5 <0.1 > 2.5000 0.874 10.023 N5 <0.1 > 2.5000 1.569 17.368 O5 <0.1 2.4076 1.115 4.263 P5 <0.1 > 2.5000 0.825 16.513 Q5 <0.1 > 2.5000 R5 <0.1 2.2046 S5 <0.1 > 2.5000 T5 <0.1 > 2.5000 U5 <1.0 > 2.5000 V5 <1.0 > 2.5000 W5 <1.0 > 2.5000

To the reaction flask, 5,7-dichloropyrazolo[1,5-a]pyrimidine (4.1 g, 22 mmol) was added along with benzyl mercaptan (2.8 mL, 22 mmol), triethylamine (3.1 mL, 22 mmol), and acetonitrile (71 mL). The reaction was stirred at room temperature for 3 hours then diluted with water, filtered and washed with water. The product, 7-(benzylthio)-5-chloropyrazolo[1,5-a]pyrimidine, was collected as a solid in 96% yield after drying under vacuum overnight.
LCMS(M+1-276)

To the reaction flask, 7-(benzylthio)-5-chloropyrazolo[1,5-a]pyrimidine (3.45 g, 12.5 mmol) was added along with 3-chloroaniline (3.3 mL, 31.3 mmol), 4N HCl in dioxane (3.1 mL, 1.2.5 mmol), and ethanol (42 mL). The reaction was stirred at reflux for 12 hours then cool to room temperature. Excess solvent was removed under vacuum and the residue was diluted with water. The mixture was made basic with 3N NaOH, filtered and washed with water. The product, 7-(benzylthio)-N-(3-chlorophenyl)pyrazolo[1,5-a]pyrimidin-5-amine, was collected, as a solid in 90% yield after drying under vacuum overnight. LCMS (M+1=376)

To 7-(benzylthio)-N-(3-chlorophenyl)pyraxolo[1,5-a]pyrimidin-5-amine (4.1. g, 11.3 mmol) in DMF (42 mL), POCl₃ (6.3 mL, 67.6 mmol) was added dropwise at room temperature. After the addition was complete, the reaction was stirred for 3 hours at room temperature. Then, the reaction was quenched by slow addition to ice cold 6N NaOH. The mixture was diluted with water and the solid was collected by filtration. The solid was washed several more times with water then dried under vacuum overnight. The product, 7-(benzylthio)-5-(3-chlorophenylamino)pyrazolo[1,5-a]pyrimidine-3-carbaldehyde, was collected as a solid in 83% yield. LCMS (M+1=395)

To the reaction flask, 7-(benzylthio)-5-(3-chlorophenylamino)pyrazolo[1,5-a]pyrimidine-3-carbaldehyde (3.7 g, 9.3 mmol) was added to ethanol (31 mL) along with hydantoin (933 mg, 9.3 mmol) and piperidine (920 µL, 9.3 mmol). The reaction was heated at 80°C for 3 days then cooled to room temperature and diluted with water. The solid was collected by filtration, washed with water, 50% ethanol/water, and then 100% ethanol. The material was dried under vacuum overnight. The product, 5-((7-(benzylthio)-5-(3-chlorophenylamino)pyrazolo[1,5-a]pyrimidin-3-yl)methylene)imidazolidine-2,4-dione, was recovered as a yellow solid in 92% yield. LCMS (M+1 = 477)

To the reaction flask, 5-((7-(benzylthio)-5-(3-chlorophenylamino)pyrazolo[1,5-a]pyimidin-3-yl)methylene)imidazolidine-2,4-dione (4.1 g, 8.6 mmol) was added to dichloromethane (86 mL) along with m-chloroperbenzoic acid (5.9g, 34.4 mmol). The mixture was allowed to stir at room temperature for 12 hours. The solid was collected by filtration, washed with dichloromethane then dried under vacuum overnight. The product, 5-((7-(benzylsulfinyl)-5-(3-chlorophenylamino)pyrazolo[1,5-a]pyrimidin-3-yl)methylene)imidaxolidine-2,4-dione, was recovered as a bright yellow solid in quantitative yield. LCMS (M+1= 493)

The compounds described in the following table were prepared using chemistries similar to those exemplified in the Examples described above. All compounds were characterized by LCMS. Table 18B shows the biological activities of the compounds listed in Table 18A. Compound CK2: IC50 (µM) PIM2: IC50 (5 µM ATP) AB: MDAMB453 (µM) AB: BxPC3 (µM) X5 <1.0 2.2449 Y5 <1.0 0.2913 Z5 <1.0 0.2968 A6 <0.1 0.209

To 5-((7-(benzylsulfinyl)-5-(3-chlorophenylamino)pyrazolo [1,5-a] pyrimidin-3-yl) methylene) imidazolidine-2,4-dione (15 mg, 0.0304 mmol) in NMP was added 2-aminoethanol (14.6 µL, 0.242 mmol). The mixture was heated in the microwave at 120 °C for 20 minutes. Water was added to the reaction mixture and the precipitate was collected by filtration. The precipitate was washed with methanol to yield 5-((5-(3-chlorophenylamino)-7-(2-hydroxyethylamino) pyrazolo [1, 5-a] pyrimidim-3-yl) methylene) imidazolidine-2,4-dione LCMS (M+1=414). Similar products (shown bellow) were also obtained as precipitates by addition of water while other reactions were purified by prep HPLC to yield corresponding products.

Same procedure as [Example 70]. LCMS (M+1=461)

Same procedure as [Example 70]. LCMS (M+1=461)

Same procedure as [Example 70]. LCMS (M+1=441)

Same procedure as [Example 70]. LCMS (M+1=412)

Same procedure as [Example 70]. LCMS (M+1=428)

Same procedure as [Example 70]. LCMS (M+1=424)

Same procedure as [Example-70]. LCMS (M+1=483)

Same procedure as [Example 70]. LCMS (M+1=461)

Same procedure as [Example 70]. LCMS (M+1=483)

Same procedure as [Example 70]. LCMS (M+1=553)

Same procedure as [Example 70]. LCMS (M+1=568)

Same procedure as [Example 70]. LCMS (M+1=452)

Same procedure as [Example 70]. LCMS (M+1=436)

Same procedure as Example 70. LCMS (M+1=439)

The compounds described in the following table were prepared using chemistries similar to those exemplified in Example 70. All compounds were characterized by LCMS. Table 19B shows the biological activities of the compounds listed in Table 19A. Compound CK2: IC50 (µM) PIM2: IC50 (5 µM ATP) AB: MDAMB453 (µM) AB: BxPC3 (µM) B6 <1.0 0.3426 10.519 >30 C6 <1.0 0.469 1.144 3.803 D6 <1.0 0.4747 0.717 1.923 E6 <1.0 1.3345 F6 <0.1 > 2.5000 G6 <1.0 > 2.5000 H6 <1.0 0.4211 I6 <1.0 0.7041 0.708 2.559 J6 <1.0 > 2.5000 K6 <1.0 0.3165 0.936 3.159 L6 <2.0 2.1905 M6 <1.0 > 2.5000 N6 <0.1 > 2.5000 O6 <0.01 > 2.5000 1.145 >30 P6 <1.0 0.7008 0.569 1.618 Q6 <1.0 1.2876 R6 <1.0 1.1213 S6 <0.1 T6 <0.1 U6 <1.0 0.5149 V6 <0.1 W6 <1.0 X6 <1.0 Y6 <0.1 Z6 <0.1 A7 <0.1

To the reaction flask, 5,7-dichloropyrazolo[1,5-a]pyrimidine (1.6 g, 8.2 mmol) was added along with tert-butyl 3-aminobenzoale (1.7 g, 8.7 mmol), triethylamine (1.2 mL, 8.6 mmol), and t-butyl alcohol (22 mL). The reaction was heated at 100°C for 6 hours then diluted with water, filtered and washed with water. The product, tert-butyl 3-(5-chloropyrazolo[1,5-a]pyrimidin-7-ylamino)benzoate, was collected as a solid in quantitative yield after drying under vacuum overnight. LCMS (M+1= 345)

To the reaction flask, tert-butyl 3-(5-chloropyrazolo[1,5-a]pyrimidin-7-ylamino)benzoate (2.9 g, 8.2 mmol) was added along with 3-chloroaniline (2.2 mL, 20.6 mmol), 4N HCl in dioxane (2.6 mL, 10.4 mmol), and t-butyl alcohol (41 mL). The reaction was stirred at 100°C for 2 days then cooled to room temperature. The mixture was diluted with water, made basic with 3N NaOH, filtered and washed with water. The product, tert-butyl 3-(5-(3-chlorophenylamino)pyrazolo[1,5-a]pyrimidin-7-ylamino)benzoate, was collected as a solid in 55% yield after drying under vacuum overnight. LCMS (M+1= 436)

To tert-butyl 3-(5-(3-chlorophenylamino)pyrazolo[1,5-a]pyrimidin-7-ylamino)benzoate (965 mg, 2.2 mmol) in DMF (8.2 mL), POCl₃ (1.2 mL, 13.3 mmol) was added dropwise at room temperature. After the addition was complete, the reaction was stirred for 3 days at room temperature. Then, the reaction was quenched by slow addition to ice cold 6N NaOH. The mixture was diluted with water and the solid was collected by filtration. The solid was washed several more times with water then dried under vacuum overnight. The product, tert-butyl 3-(5-(3-chlorophenylamino)-3-formylpyrazolo[1,5-a]pyrimidin-7-ylamino)benzoate, was collected as a solid in 12% yield after purification by column chromatography on silica using 5% acetone/dichloromethane as the eluent. LCMS (M+1= 464)

To the reaction flask, tert-butyl. 3-(5-(3-chlorophenylamino)-3-formylpyrazolo[1,5-a]pyrimidin-7-ylamino)benzoate (122 mg, 0.3 mmol) was added to ethanol (1.3 mL) along with hydantoin (26 mg, 0.3 mmol) and piperidine (26 µL, 0.3 mmol). The reaction was heated at 80°C for 2 hours in the microwave then cooled to room temperature and diluted with water. The solid was collected by filtration, washed with water, 50% ethanol/water, and then 100% ethanol. The material was dried under vacuum overnight. The product, tert-butyl 3-(5-(3-chlorophenylamino)-3-((2,5-dioxoimidazolidin-4-ylidene)methyl)pyrazolo[1,5-a]pyrimidin-7-ylamino)benzoate, was recovered as a solid in 69% yield. LCMS (M+1= 546)

Tert-butyl 3-(5-(3-chlorophenylamino)-3-((2,5-dioxoimidazolidin-4-ylidene)methyl)pyrazolo[1,5-a]pyrimidin-7-ylamino)benzoate (97 mg, 0.2 mmol) was dissolved in 2 mL of TFA/DCM (1:1) and stirred at room temperature for 1 hour. Excess solvent and TFA were removed by evaporation under a stream of nitrogen. The residue was diluted with water then the mixture was filtered. The product, 3-(5-(3-chlorophenylamino)-3-((2,5-dioxoimidazolidin-4-ylidene)methyl)pyrazolo[1,5-a]pyrimidin-7-ylamino)benzoic acid, was collected as a solid in 85% yield. LCMS (M+1= 490)

To the reaction flask, 3-(5-(3-chlorophenylamino)-3-((2,5-dioxoimidazolidin-4-ylidene)methyl)pyrazolo[1,5-a]pyrimidin-7-ylamino)benzoic acid (30 mg, 0.06 mmol) was added to DMF (0.5 mL) along with HOBt (9.2 mg, 0.06 mmol), triethylamine (8.4 µL, 0.06 mmol) and tert-butyl piperazine-1 -carboxylate (11.2 mg, 0.06 mmol). The reaction mixture was stirred at room temperature for 5 minutes then EDC (11.5 mg, 0.06 mmol) was added. The reaction was allowed to stir for an additional hour then diluted with water and filtered. The recovered solid was washed with more water followed by ethanol. The product, tert-butyl 4-(3-(5-(3-chlorophenylamino)-3-((2,5-dioxoimidazolidin-4-ylidene)methyl)pyrazolo[1,5-a]pyrimidin-7-ylamino)benzoyl)piperazine-1-carboxylate, was collected as a solid in 73% yield.
LCMS (M+1 = 658)

Tert-butyl 4-(3-(5-(3-chlorophenylamino)-3-((2,5-dioxoimidazolidin-4-ylidene)methyl)pyrazolo[1,5-a]pyrimidin-7-ylamino)benzoyl)piperazine-1-carboxylate (27 mg, 0.04 mmol) was dissolved in 2 mL of TFA/DCM (1:1) and stirred at room temperature for 1 hour. Excess solvent and TFA were removed by evaporation under a stream of nitrogen. The residue was diluted with water then the mixture was filtered. The recovered solid was washed with water followed by 50% ethanol. The product, 5-((5-(3-chlorophenylamino)-7-(3-(piperazine-1-carbonyl)phenylamino)pyrazolo[1,5-a]pyrimidin-3-yl)methylene)imidazolidine-2;4-dione, was collected as a solid in 21% yield. LCMS (M+1 = 558)

Same procedure as [Example 90]. LCMS (M+1=586)

The compounds described in the following table were prepared using chemistries similar to those exemplified in the Examples described above. All compounds were characterized by LCMS. Table 20B shows the biological activities of the compounds listed in Table 20A. Compound CK2: IC50 (µM) PIM2: IC50 (5 µM ATP) AB: MDAMB453 (µM) AB: BxPC3 (µM) B7 <0.01 > 2.5000 7.773 10.204 C7 <0.01 1.4446 6.435 > 30 D7 <0.1 > 2.5000 14.229 > 30 E7 <0.01 2.0193 0.364 3.006 F7 <0.01 1.2348 1.587 13.969

To the reaction flask, 5,7-dichloropyrazolo[1,5-a]pyrimidine (452 mg, 2.4 mmol) was added along with (3-cyanobenzyl)zinc(II) bromide (6 mL, 3.75 mmol, 0.625M in DMF), Pd(PPh₃)₄ (110 mg, 0.1 mmol), and DMF (10 mL). The reaction was heated at 60°C for 4 hours then cooled to room temperature. The reaction mixture was poured into saturated aqueous NH₄Cl solution and ice and extracted with ethyl acetate. The combined extracts were washed with water, saturated NaCl solution, and then dried over Na₂SO₄. The solvent was removed in vacuo and the residue was purified by column chromatography on silica using 35% ethyl acetate/hexanes as the eluent. The product, 3-((7-chloropyrazolo[1,5-a]pyrimidin-5-yl)methyl)benzonitrile, was recovered in 64% yield. LCMS (M+1= 269)

To the reaction flask, 3-((7-chloropyrazolo[1,5-a]pyrimidin-5-yl)methyl)benzonitrile (400 mg, 1.5 mmol) was added along with cyclopropylamine (115 µL, 1.6 mmol), triethylamine (230 µL, 1.6 mmol), and acetonitrile (3 mL). The reaction was stirred at room temperature for 8 hours at 80°C then cooled to room temperature, diluted with water, filtered and washed with water. The product, 3-((7-(cyclopropylamino)pyrazolo[1,5-a]pyrimidin-5-yl)methyl)benzonitrile, was collected as a solid in 83% yield after drying under vacuum overnight. LCMS (M+1=290)

To 3-((7-(cyclopropylamino)pyrazolo[1,5-a]pyrimidin-5-yl)methyl)benzonitrile (69 mg, 0.24 mmol) in DMF (0.6 mL), POCl₃ (130 µL, 1.4 mmol) was added at room temperature. After the addition was complete, the reaction was stirred for 1 hour at room temperature. Then, the reaction was quenched by addition to ice cold 6N NaOH. The mixture was diluted with water and the solid was collected by filtration. The solid was washed several more times with water then dried under vacuum overnight. The product, 3-((7-(cyclopropylamino)-3-formylpyrazolo[1,5-a]pyrimidin-5-yl)methyl)benzonitrile, was collected as a solid in 37% yield. LCMS (M+1= 318)

To the reaction flask, 3-((7-(cyclopropylamino)-3-formylpyrazolo[1,5-a]pyrimidin-5-yl)methyl)benzonitrile (28 mg, 0.09 mmol) was added to ethanol (0.5 mL) along with hydantoin (9 mg, 0.09 mmol) and piperidine (9 µL, 0.09 mmol). The reaction was heated at 80°C for 30 minutes in the microwave then cooled to room temperature and diluted with water. The solid was collected by filtration, washed with water, 50% ethanol/water, and then 1000% ethanol. The material was dried under vacuum overnight. The product, 3-((7-(cyclopropylamino)-3-((2,5-dioxoimidazolidin-4-ylidene)methyl)pyrazolo[1,5-a]pyrimidin-5-yl)methyl)benzonitrile, was recovered as a solid in 34% yield. LCMS (M+1 = 400) Structure LCMS m/z [M+1]+ CK2: IC50 (uM) PIM2: IC50 (Sum ATP) AB: MDAMB453 (uM) AB: BxPC3 (uM) 400 <0.01 > 2.5000 > 30 19.66

To tert-butyl. 5-chloro-3-formylpyrazolo[1,5-a]pyrimidin-7-yl (cyclopropyl)carbamate (650 mg, 1.93 mmol) in 14 mL of a 2:1 mixture of 1,2-dimethoxyethane/EtOH was added 3-hydroxyphenyl boronic acid (399 mg, 2.89 mmol), tetrakis(triphenylphosphine)palladium(0) (112 mg, 0.096 mmol), and 2M aqueous solution of Na₂CO₃ (2.9 mL, 5.79 mmol). The mixture was stirred at 85°C for 1h. The volatiles were removed by rotary evaporation and the residue was purified by silica gel chromatography (0%-30% EtOAc/Hexanes) to provide 400 mg of tert-butyl cyclopropyl(3-formyl-5-(3-hydroxyphenyl)pyrazolo[1,5-a]pyrimidin-7-yl)carbamate. (52%). LCMS (M+1=395)

To tert-butyl cyclopropyl(3-formyl-5-(3-hydroxyphenyl)pyrazolo[1,5-a]pyrimidin-7-yl)carbamate (400 mg, 1.01 mmol) in methylene chloride (20 mL) was added TFA (10 mL). The reaction mixture was stirred at room temperature for 2h. The volatiles were removed by rotary evaporation and the residue was purified by silica gel chromatography (0%-40% EtOAc/hexanes) to provide 103 mg of 7-(cyclopropylamino)-5-(3hydroxyphenyl)pyrazolo [1,5-a]pyrimidine-3-carbaldehyde. (35%). LCMS (M+1=295)

To 7-(cyclopropylamino)-5-(3hydroxyphenyl)pyrazolo [1,5-a]pyrimidine-3-carbaldehyde (100 mg, 0.34 mmol) in EtOH (2 mL) was added piperidine (67 µL, 0.68 mmol), and Hydantoin (34 mg, 0.34 mmol)). The reaction was stirred at 50°C overnight. The solid formed was isolated by filtration to provide 70 mg of 5-((7-(cyclopropylamino)-5-(3-hydroxyphenyl)pyrazolo[1,5-a]pyrimidin-3-yl)methylene)imidazolidine-2,4-dione (55%). LCMS (M+1=377)

Same procedure as [Example 97]. LCMS (M+1=463)

Same procedure as [Example 98]. LCMS (M+1=363)

Same procedure as [Example 99]. LCMS (M+1=445)

Same procedure as [Example 97]. LCMS (M+1 = 409)

Same procedure as [Example 98]. LCMS (M+1= 309)

Same procedure as [Example 99]. LCMS (M+1= 391)

Same procedure as [Example 97]. LCMS (M+1= 437)

Same procedure as [Example 98]. LCMS (M+1= 337)

Same procedure as [Example 99]. LCMS (M+1=419)

Same procedure as [Example 97] . LCMS (M+1= 457)

Same procedure as [Example 98]. LCMS (M+1= 357)

Same procedure as [Example 99]. LCMS (M+1= 439)

Same procedure as [Example 97]. LCMS (M+1= 472)

Same procedure as [Example 98]. LCMS (M+1= 372)

Same procedure as [Example 99]. LCMS (M+1= 454)

Same procedure as [Example 97]. LCMS (M+1= 472)

Same procedure as Example 98]. LCMS (M+1= 372.)

Same procedure as [Example 99]. LCMS (M+1 = 454)

Same procedure as [Example 97]. LCMS (M+1= 422)

Same procedure as [Example 98]. LCMS (M+1= 322.)

Same procedure as [Example 99]. LCMS (M+1= 404)

Same procedure as [Example 97]. LCMS (M+1= 404)

Same procedure as [Example 98]. LCMS (M+1= 304)

Same procedure as [Example 99]. LCMS (M+1=386)

Same procedure as [Example 99]. LCMS (M+1= 397)

Same procedure as [Example 98]. LCMS (M+1= 297)

Same procedure as [Example 99]. LCMS (M+1= 379)

Same procedure as [Example 97]. LCMS (M+1= 380)

Same procedure as [Example 98]. LCMS (M+1=280)

Same procedure as [Example 99]. LCMS (M+1= 362)

Same procedure as [Example 97]. LCMS (M+1=380)

Same procedure as [Example 98]. LCMS (M+1= 280)

Same procedure as [Example 99]. LCMS (M+1= 362)

Same procedure as [Example 97]. LCMS (M+1 = 398)

Same procedure as [Example 98]. LCMS (M+1=298)

Same procedure as [Example 99]. LCMS (M+1= 380)

Same procedure as [Example 97]. LCMS (M+1=395)

Same procedure as [Example 98]. LCMS (M+1=295)

Same procedure as [Example 99]. LCMS (M+1=377)

Same procedure as [Example 97]. LCMS (M+1= 478)

Same procedure as [Example 98]. LCMS (M+1= 378)

Same procedure as [Example 99]. LCMS (M+1= 460)

Same procedure as [Example 97]. LCMS (M+1=491)

Same procedure as [Example 98]. LCMS (M+1=391)

Same procedure as [Example 99]. LCMS (M+1= 473)

Same procedure as [Example 97]. LCMS (M+1= 450)

Same procedure as [Example 98]. LCMS (M+1 = 350)

Same procedure as [Example 99]. LCMS (M+1= 432)

To tert-butyl 5-(3-(acetamidomethyl)phenyl)-3-formylpyrazolo[1,5-a]pyrimidin-7-yl(cyclopropyl)carbamate (50 mg, 0.111 mmol) was added 1 mL of 4M HCl in 1,4-dioxane and 1 ml of H₂O. The reaction mixture was stirred at 80° C for 16 hours then cooled to room temperature and diluted with H₂O. To the reaction mixture, 5M NaOH was added to adjust pH to >10 then the mixture was extracted with CH₂Cl₂. The organic layer was collected, dried over MgSO₄, filtered and evaporated to dryness to provide 24 mg of 5-(3-(aminomethyl)phenyl)-7-(cyclopropylamino)pyrazolo[1,5-a]pyrimidine-3-carbaldehyde (70%). LCMS (M+1=308)

Same procedure as [Example 99]. LCMS (M+1 = 390)

The compounds described in the following table were prepared using chemistries similar to those exemplified in Example 98 and Example 99. All compounds were characterized by LCMS. Table 22B shows the biological activities of the compounds listed in Table 22A. Compound CK2: IC50 (µM) PIM2: IC50 (5 µM ATP) AB: MDAMB453 (µM) AB: BxPC3 (µM) G7 <0.01 0.6135 0.33 10.627 H7 <0.01 0.8908 0.402 1.541 I7 <0.01 > 2.5000 3.743 3.68 J7 <0.01 0.6492 0.477 5.171 K7 <0.01 > 2.5000 1.709 2.054 L7 <0.01 > 2.5000 0.517 13.111 M7 <0.1 > 2.5000 N7 <0.01 2.2144 0.284 2.916 O7 <0.01 > 2.5000 > 30 > 30 P7 <0.01 2.1685 7.866 7.907 Q7 <0.1 2.4032 1.494 3.279 R7 <0.01 > 2.5000 1.054 23.617 S7 <0.01 > 2.5000 1.174 11.78 T7 <0.01 > 2.5000 1.298 6.592 U7 <0.01 > 2.5000 1.153 1.191 V7 <0.01 > 2.5000 1.964 14.486 W7 <0.1 > 2.5000 0.683 1.898 X7 <0.01 0.867 4.746 > 30 Y7 <0.1 1.3082 1.938 2.578 Z7 <0.01 1.4748 1.79 0.725 A8 <0.01 1.2497 > 30 14.347 B8 <0.01 > 2.5000 > 30 12.535 C8 <0.01 > 2.5000 17.123 1.232 D8 <0.01 0.0754 5.276 0.549 E8 <0.01 > 2.5000 11.733 > 30 F8 <0.01 0.2562 1.068 0.745 G8 <0.01 0.0487 14.882 10.61 H8 <0.01 > 2.5000 20.012 4.608 I8 <0.1 > 2.5000 1.706 2.744 J8 <0.01 > 2.5000 1.263 8.129 K8 <0.1 > 2.5000 12.417 > 30 L8 <0.01 2.084 12.278 > 30 M8 <0.01 1.7271 > 30 > 30 N8 <0.01 > 2.5000 1.979 2.253 O8 <0.01 > 2.5000 15.69 29.035 P8 <0.01 0.948 1.742 Q8 <0.01 > 2.5000 26.74 5.426 R8 <1.0 > 2.5000

Same procedure as [Example 97]. LCMS (M+1= 443)

Same procedure as [Example 98]. LCMS (M+1=343)

Same procedure as [Example 99]. LCMS (M+1=425)

Same procedure as [Example 97]. LCMS (M+1= 410)

Same procedure as [Example 98]. LCMS (M+1= 310)

Same procedure as [Example 99]. LCMS (M+1 = 392)

To tert-butyl 5-chloro-3-formylpyrazolo[1,5-a]pyrimidin-7-yl (cyclopropyl)carbamate (1 g, 2.97 mmol) in 30 mL of a 2:1 mixture of 1,2-dimethoxyethane/ EtOH was added 2-carboxythiophene-5- boronic acid (766 mg, 4.45 mmol), tetrakis(triphenylphosphine)palladium(0) (171 mg, 0.148 mmol), and 2M aqueous solution of Na₂CO₃ (4.45 mL, 8.91 mmol). The mixture was stirred at 95°C for 3 hours then cooled to room temperature and partitioned between 2N NaOH and ethyl acetate. The layers were separated and the aqueous layer was acidified to pH<3 with conc. HCl. The aqueous layer was extracted (3x) with methylene chloride. The combined organic layers was washed with brine, dried over MgSO₄, filtered, and evaporated to dryness to provide 450 mg of 5-(7-(tert-butoxycarbonyl(cyclopropyl)amino)-3-formylpyrazolo[1,5-a]pyrimidin-5-yl)thiophene-2-carboxylic acid. Some additional material which was in the first ethyl acetate layer was purified by silica gel chromatography (0%-20% MeOH/CH₂Cl₂) to provide another 550 mg of 5-(7-(tert-butoxycarbonyl(cyclopropyl)amino)-3-formylpyrazolo[1,5-a]pyrimidin-5-yl)thiophene-2-carboxylic acid (79%). LCMS (M+1=429)

To 5-(7-(tert-butoxycarbonyl(cyclopropyl)amino)-3-formylpyrazolo[1,5-a]pyrimidin-5-yl)thiophene-2-carboxylic acid (1 g, 2.33 mmol) was added 8 mL of 4M HCl in dioxane and another 5 mL of dioxane. The reaction mixture was stirred at 80° C for 2 hours, cooled to room temperature and partitioned between CH₂Cl₂ and H₂O. The emulsion that formed between the layers was filtered off and rinsed with H₂O. The recovered solid was dried under vacuum to provide 627 mg of 5-(7-(cyclopropylamino)-3-formylpyrazolo[1,5-a]pyrimidin-5-yl)thiophene-2-carboxylic acid as a red solid (82%). LCMS (M+1=329)

To 5-(7-(cyclopropylamino)-3-formylpyrazolo[1,5-a]pyrimidin-5-yl)thiophene-2-carboxylic acid (30 mg, 0.091 mmol), EDCI (19 mg, 0.10 mmol), Et₃N (14 µL, 0.10 mmol), and HOBt (14 mg, 0.10 mmol) in 2 mL of DMF pre-stirred for 5 minutes was added 3-methoxypropylamine (10 µL, 0.10 mmol). The reaction mixture was stirred at room temperature for 1 hour. The reaction was diluted with ethyl acetate, washed with H₂O, brine, dried over MgSO₄, filtered, and evaporated to dryness to provide 30 mg of 5-(7-(cyclopropylamino)-3-formylpyrazolo[1,5-a]pyrimidin-5-yl)-N-(3-methoxypropyl)thiophene-2-carboxamide (83%). LCMS (M+1=400)

To 5-(7-(cyclopropylamino)-3-formylpyrazolo[1,5-a]pyrimidin-5-yl)-N-(3-methoxypropyl)thiophene-2-carboxamide (30 mg, 0.075 mmol) in EtOH (1 mL) was added piperdine (20 µL, 0.150 mmol), and hydantoin (10 mg, 0.075 mmol). The reaction mixture was stirred at 85°C for 3 hours. The solid formed was isolated by filtration to provide 5-(7-(cyclopropylamino)-3-((2,5-dioxoimidazolidin-4-ylidene)methyl)pyrazolo[1,5-a]pyrimidin-5-yl)-N-(3-methoxypropyl)thiophene-2-carboxamide. LCMS (M+1=482)

To 5-(7-(cyclopropylamino)-3-formylpyrazolo[1,5-a]pyrimidin-5-yl)thiophene-2-carboxylic acid (40 mg, 0.122 mmol.), HATU (70 mg, 0.183 mmol), HOBt (4mg, 0.024 mmol) and DIEA (85 µL, 0.488 mmol) in 2 mL of DMF was added ammonium chloride (20 mg, 0.366 mmol). The reaction mixture was stirred at room temperature for 1 hour. The reaction mixtue was diluted with ethyl acetate washed with saturated NaHCO₃ solution, brine, dried over MgSO4, filtered, and evaporated to dryness to provide 42 mg of 5-(7-(cyclopropylamino)-3-formylpyrazolo[1,5-a]pyrimidin-5-yl)thiophene-2-carboxamide (100%). LCMS (M+1=328).

Same procedure as [Example 159]. LCMS (M+1=410)

The compounds described in the following table were prepared using chemistries similar to those exemplified in Example 158 and Example 159. All compounds were characterized by LCMS. Table 23B shows the biological activities of the compounds listed in Table 23A. Compound CK2: IC50 (µM) PIM2: IC50 (5 µM ATP) AB: MDAMB453 (µM) AB: BxPC3 (µM) S8 <0.01 1.1336 0.648 0.99 T8 <0.01 > 2.5000 0.141 0.837 U8 <0.01 1.0091 10.145 6.668 V8 <0.01 > 2.5000 0.416 > 30 W8 <0.01 > 2.5000 0.425 0.417 X8 <0.01 > 2.5000 0.689 > 30 Y8 <0.01 > 2.5000 0.358 0.642 Z8 <0.01 > 2.5000 0.162 1.15 A9 <0.01 > 2.5000 0.542 1 B9 <0.01 > 2.5000 0.49 3.925 C9 <0.01 > 2.5000 0.171 0.822 D9 <0.01 > 2.5000 0.869 1.983 E9 <0.01 > 2.5000 0.397 0.496 F9 <0.01 > 2.5000 0.312 0.643 G9 <0.01 > 2.5000 0.31 0.657 H9 <0.01 > 2.5000 0.251 10.512 I9 <0.01 0.7137 > 30 > 30 J9 <0.01 > 2.5000 0.795 1.736 K9 <0.01 > 2.5000 9.378 11.666 L9 <0.01 > 2.5000 2.066 3.829 M9 <0.01 > 2.5000 1.266 1.469 N9 <0.01 1.134 5.413 O9 <0.01 0.621 12.558 P9 <0.01 0.596 0.5 Q9 <0.01 1.044 2.134 R9 <0.01 1.554 1.555 S9 <0.01 5.882 5.532 T9 <0.01 0.444 0.956 U9 <0.01 1.479 4.863 V9 <0.01 1.567 2.905 W9 <0.01 1.145 0.885 X9 <0.01 > 2.5000 1.391 > 30 Y9 <0.01 > 2.5000 0.389 0.438 Z9 <0.01 > 2.5000 0.762 1.337 A10 <0.01 > 2.5000 0.408 2.115 B10 <0.01 > 2.5000 0.895 1.167 C10 <0.01 0.7939 0.66 2.399 D10 <0.01 > 2.5000 1.529 6.508 E10 <0.01 > 2.5000 0.557 0.624 F10 <0.01 > 2.5000 0.251 0.323 G10 <0.01 > 2.5000 1.038 0.995 H10 <0.01 > 2.5000 0.294 2.968 I10 <0.01 > 2.5000 0.813 1.386 J10 <0.01 > 2.5000 0.613 0.324 K10 <0.01 > 2.5000 0.579 0.451 L10 <0.01 > 2.5000 2.275 0.792 M10 <0.01 1.7758 5.94 0.677 N10 <0.01 > 2.5000 0.958 0.455 O10 <0.01 1.8944 0.537 0.297 P10 <0.01 > 2.5000 0.394 0.451 Q10 <0.01 > 2.5000 1.782 16.637 R10 <0.01 > 2.5000 1.641 5.729 S10 <0.01 > 2.5000 10.053 18.645 T10 <0.1 > 2.5000 > 30 >30 U10 <0.01 > 2.5000 13.297 21.203 V10 <0.01 2.1321 > 30 >30 W10 <0.01 1.3653 0.236 0.63 X10 <0.01 > 2.5000 0.937 0.917 Y10 <0.01 > 2.5000 0.79 > 30 Z10 <0.01 > 2.5000 2.336 22.798 A11 <0.01 > 2.5000 0.458 0.724 B11 <0.01 > 2.5000 > 30 1.262 C11 <0.0.1 > 2.5000 27.783 3.302 D11 <0.01 > 2.5000 1.445 2.265 E11 <0.01 > 2.5000 1.298 2.948 F11 <0.01 > 2.5000 0.567 0.903 G11 <0.01 2.0441 0.231 0.494 H11 <0.01 > 2.5000 1.11 2.705 I11 <0.01 > 2.5000 1.232 0.591 J11 <0.01 > 2.5000 0.833 1.234 K11 <0.01 > 2.5000 0.546 1.257 L11 <0.01 > 2.5000 1.004 0.816 M11 <0.01 > 2.5000 1.016 0.745 N11 <0.01 > 2.5000 1.266 2.261 O11 <0.01 > 2.5000 0.887 4.986 P11 <0.01 > 2.5000 0.487 0.517 Q11 <0.01 > 2.5000 0.621 0.564 R11 <0.01 > 2.5000 0.845 2.309 S11 <0.01 > 2.5000 1.935 >30 T11 <0.01 > 2.5000 0.193 >30 U11 <0.01 > 2.5000 0.618 5.349 V11 <0.01 > 2.5000 0.892 1.6 W11 <0.01 > 2.5000 0.156 3.435 X11 <0.01 1.7245 3.806 0.225 Y11 <0.01 > 2.5000 1.402 0.352 Z11 <0.01 1.2434 2.251 0.355 A12 <0.01 1.4396 1.151 0.445 B12 <0.01 > 2.5000 0.399 2.764 C12 <0.01 > 2.5000 >30 >30 D12 <0.01 > 2.5000 0.683 0.854 E12 <0.01 > 2.5000 29.518 2.348 F12 <0.01 > 2.5000 >30 >30 G12 <0.01 0.8106 0.658 0.352 H12 <0.01 > 2.5000 0.449 0.418 I12 <0.01 > 2.5000 1.282 1,516 J12 <0.01 > 2.5000 0.52 0.94 K12 <0.01 1.872 1.338 0.379 L12 <0.01 > 2.5000 0.498 >30 M12 <0.01 1.2604 7.403 8.736 N12 <0.01 3.52 >30 O12 <0.01 > 2.5000 1.077 2.509 P12 <0.01 > 2.5000 1.014 3.421 Q12 <0.01 > 2.5000 0.942 7.084 R12 <0.01 > 2.5000 0.846 14.096 S12 <0.01 > 2.5000 1.034 4.897 T12 <0.01 > 2.5000 0.767 2.662 U12 <0.01 > 2.5000 0.525 > 30 V12 <0.01 1.759 > 30 W12 <0.01 1.041 1.184 X12 <0.01 7.54 > 30 Y12 <0.01 0.692 1.706 Z12 <0.01 2.17 9.892 A13 <0.01 > 2.5000 0.534 0.996 B13 <0.01 > 2.5000 0.388 2.584

Tert-butyl 5-chloro-3-formylpyrazolo[1,5-a]pyrimidine-7-yl)(cyclopropyl)carbamate (0.5 g, 1.48 mmol) and commercially available (Combi-Blocks) 2-carboxythiophene-4-boronic acid pinacol ester (754 mg, 2.97 mmol) were dissolved in acetonitrile. 2M Na₂CO₃ (1 mL) was added and the solution was degassed with a stream of N₂ for 10 min. PdCl₂dppf·CH₂Cl₂ (60 mg, 0.07 mmol) was added and the reaction was heated to 100°C for 1.5 h. The solution was diluted with 1.5N NaOH (80 mL) and filtered over celite. The pH of the filtrate was adjusted to pH=3 by the addition of 6M HCl. The resulting precipitate was filtered and dried in vacuo to afford 4-(7-(tert-butoxycarbonyl(cyclopropyl)amino)-3-formylpyrazolo[1,5-a]pyrimidin-5-yl)thiophene-2-carboxylic acid (473 mg, 74%) as a tan solid. LCMS (ES): >90% pure, m/z 429 [M+1]⁺.

4-(7-(Tert-butoxycarbonyl(cyclopropyl)amino)-3-formylpyrazolo[1,5-a]pyrimidin-5-yl)thiophene-2-carboxylic acid (473 mg, 1.10 mmol) was dissolved in dichloromethane (5 mL) and trifluoroacetic acid (3 mL). After 1 h, the dark red solution was concentrated under a stream of air. The red oil was triturated with Et₂O (5 mL) and the precipitate was filtered to provide 4-(7-(cyclopropyl)amino)-3-formylpyrazolo[1,5-a]pyrimidin-5-yl)thiophene-2-carboxylic acid (321 mg, 88%). LCMS (ES): >95% pure, m/z 329 [M+1]⁺.

Hydantoin (292 mg, 2.92 mmol) and piperidine (285 µL, 2.89 mmol) were added to 4-(7-(cyclopropyl)amino)-3-formylpyrazolo[1,5-a]pyrimidin-5-yl)thiophene-2-carboxylic acid (315 mg, 0.96 mmol) dissolved in ethanol (5 mL). The reaction was heated at 80°C. After 15 h, the reaction was cooled to r.t., then diluted with water (10 mL). The pH was adjusted to pH=3 by addition of 1N HCl. The yellow precipitate was collected and washed with 1:1 ethanol:water (10 mL) and then ethanol (10 mL). The solid was dried in vacuo to give (Z)-4-(7-(cyclopropyl)amino)-3-((2,5-dioxoimidazolidin-4-ylidene)methyl)pyrazolo[1,5-a]pyrimidin-5-yl)thiophene-2-carboxylic acid (362 mg, 92%). LCMS (ES): >95% pure, m/z 411 [M+1]⁺.

(Z)-4-(7-(Cyclopropyl)amino)-3-((2,5-dioxoimidazolidin-4-ylidene)methyl)pyrazolo[1,5-a]pyrimidin-5-yl)thiophene-2-carboxylic acid (1.0 eq, 34 mg, 0.0828 mmol) was mixed in a vial with HOBt.H₂O (2.0 eq, 22 mg, 0.163 mmol), 2,6 dimethylmorpholine (isomer mixture, 4.0 eq, 41 ul, 0.333 mmol), DIEA (2.0 eq, 29 ul, 0.166 mmol) in NMP (0.5 ml). EDCI (2.0 eq, 32 mg, 0.166 mmol) was added and the mixture was stirred at 70°C for 1 hour. Water was added and the resulting precipitate was filtered and dried. The material was triturated in a mixture of ethyl acetate and hexanes, filtered and dried in vacuo to give (Z)-5-((7-(cyclopropylamino)-5-(5-(2,6-dimethylmorpholine-4-carbonyl)thiophen-3-yl)pyrazolo[1,5-a]pyrimidin-3-yl)methylene)imidazolidine-2,4-dione as a yellow solid (26 mg, 62% yield). LCMS (ES): >95% pure, m/z 508 [M+1]⁺.

The following compounds were prepared using conditions similar to the chemistries described in Example 165. All compounds were characterized by LCMS. Table 24B shows the biological activities of the compounds listed in Table 24A. Compound CK2: IC50 (µM) PIM2: IC50 (5 µM ATP) AB: MDAMB453 (µM) AB: BxPC3 (µM) C13 <0.01 2.382 >30 >30 D13 <0.01 > 2.5000 0.605 0.591 E13 <0.01 0.982 1.469 3.466 F13 <0.01 0.6084 1.641 0.943 G13 <0.01 0.888 0.845 1.251 H13 <0.01 0.654 3.98 10.149 I13 <0.01 1.8781 15.19 > 30 J13 <0.01 0.548 0.266 1.348 K13 <0.01 > 2.5000 4.31 9.291 L13 <0.01 1.9547 1.548 0.767 M13 <0.01 > 2.5000 10.179 4.429 N13 <0.01 1.9848 3.335 4.142 O13 <0.01 > 2.5000 6.095 19.358 P13 <0.01 0.8133 2.772 8.499 Q13 <0.01 >30 6.578 R13 <0.01 > 2.5000 1.657 2.293

5-(Hydroxymethyl)thiophen-2-boronic acid was prepared from the commercially available 5-formylthiophen-2-boronic acid (Combi-Blocks) according to the procedure described in patent application WO2007/118137.

Note: DME and 2M Na₂CO₃ were degassed with a stream of N₂ in separate flasks prior to addition. Tert-butyl 5-chloro-3-formylpyrazolo[1,5-a]pyrimidine-7-yl)(cyclopropyl)carbamate (1.5 g, 4.45 mmol) was dissolved in DME (40 mL). Crude 5-(hydroxymethyl)thiophen-3-boronic acid (1.4 g, 8.9 mmol) was added, followed by Pd(PPh₃)₄ (510 mg, 0.45 mmol) and finally 2M Na₂CO₃ (6.7 ml, 13.3 mmol). The reaction was heated to 90°C for 2 h. The solution was partitioned between EtOAc (100 mL) and 0.5N HCl (100 mL). The aqueous layer was extracted with EtOAc (2 x 75 mL). The organics were washed with brine (250 mL), dried over MgSO₄ filtered and concentrated in vacuo. The residue was purified via flash column chromatography (30-45% EtOAc/hexanes) and then triturated with hexanes (3 x 10 mL) to yield tert-butyl cyclopropyl(3-formyl-5-(5-(hydroxymethyl)thiophen-2-yl)pyrazolo[1,5-a]pyrimidine-7-yl)carbamate (984 mg, 53%) as an off white solid. LCMS (ES): >95% pure, m/z 415 [M+1]⁺.

Hydrogen bromide (48% in water, 5 mL) was added dropwise to tert-butyl cyclopropy)(3-formyl-5-(5-(hydroxymethyl)thiophen-2-yl)pyrazolo[1,5-a]pyrimidine-7-yl)carbamate (980 mg, 2.4 mmol) suspended in dichloromethane (5 mL). The solution immediately became dark brown and homogeneous upon addition. The reaction was heated to 40°C for 4 hours, then diluted with dichloromethane (10 mL). The liquid was decanted and the gummy residue was washed with dichloromethane (3x10 mL). The combined liquids were washed successively with sat. NaHCO₃ (20 mL) and brine (20 mL), and then dried over MgSO₄, filtered and concentrated in vacuo. The residue was triturated with hexanes and then purified via flash column chromatography (10-20% EtOAc/hexanes) to provide 5-(5-(bromomethyl)thiophen-2-yl)-7-(cyclopropylamino)pyrazolo[1,5-a]pyrimidine-3-carbaldehyde (300 mg, 34%) as a yellow solid. LCMS (ES): >95% pure, m/z 378 [M+1]⁺.

Potassium carbonate (30 mg, 0.20 mmol) was added to 5-(5-(bromomethyl)thiophen-2-yl)-7-(cyclopropylamino)pyrazolo[1,5-a]pyrimidine-3-carbaldehyde (25 mg, 0.07 mmol) dissolved in DMF (0.7 mL). Pyrrolidone (6 µL, 0.07 mmol) was added and the reaction was heated to 60°C for 4 h. Water (3 mL) was added and the orange precipitate was filtered and dried in vacuo to give 7-(cyclopropylamino)-5-(5-(pyrrolidin-1-ylmethyl)thiophen-2-yl)pyrazolo[1,5-a]pyrimidine-3-carbaldehyde (13 mg, 54%) which was used without further purification. LCMS (ES): >85% pure, m/z 368 [M+1]⁺.

Hydantoin (3 mg, 0.03 mmol) and piperidine (3 µL, 0.03 mmol) were added to 7-(cyclopropylamino)-5-(5-(pyrrolidin-1-ylmethyl)thiophen-2-yl)pyrazolo[1,5-a]pyrimidine-3-carbaldehyde (12 mg, 0.03 mmol) dissolved in ethanol (0.5 mL). The reaction was heated at 80 °C. After 15h, the reaction was cooled to room temperature then diluted with water (3 mL). The precipitate was collected and washed with 1:1 ethanol:water (3 mL) and dried in vacuo to furnish (Z)-5-((7-(cyclopropylamino)-5-(5-pyrrolidin-1-ylmethyl)thiophen-2-yl)pyrazolo[1,5-a]pyrimidin-3-yl)methylene)imidazolidine-2,4-dione (2.8 mg, 9% over two steps). LCMS (ES): >95% pure, m/z 450 [M+1]⁺.

The compounds described in the following table were prepared using chemistries similar to those exemplified in Example 168 and Example 169. All compounds were characterized by LCMS. Table 25B shows the biological activities of the compounds listed in Table 25A. Compound CK2: IC50 (µM) PIM2: IC50 (5 µM ATP) AB: MDAMB453 (µM) AB: BxPC3 (µM) S13 <0.01 > 2.5000 1.266 2.261 T13 <0.01 0.8106 0.658 0.352 U13 <0.01 > 2.5000 0.449 0.418 V13 <0.01 > 2.5000 1.282 1.516 W13 <0.01 > 2.5000 0.52 0.94 X13 <0.01 1.872 1.338 0.379

5-(Hydroxymethyl)thiophen-3-boronic acid was prepared from the commercially available 5-formylthiophen-3-boronic acid (Combi-Blocks) according to the procedure described in patent application WO2007/118137.

Note: DME and 2M Na₂CO₃ were degassed with a stream of N₂ in separate flasks prior to addition. Tert-butyl 5-chloro-3-formylpyrazolo[1,5-a]pyrimidine-7-yl)(cyclopropyl)carbamate (750 mg, 2.22 mmol) was dissolved in DME (22 mL). Crude 5-(hydroxymethyl)thiophen-3-boronic acid (880 mg, 5.57 mmol) was added, followed by Pd(PPh₃)₄ (256 mg, 0.22 mmol) and finally 2M Na₂CO₃ (3.3 mL, 6.60 mmol). The reaction was heated to 90°C for 2h. The solution was partitioned between EtOAc (100 mL) and 0.5N HCl (100 mL). The aqueous layer was extracted with EtOAc (2 x 75 mL). The organics were washed with brine (250 mL), dried over MgSO₄, filtered and concentrated in vacuo. The residue was purified via flash column chromatography (30-45% EtOAc/hexanes) and then triturated with hexanes (3 x 10 mL) to yield tert-butyl cyclopropyl(3-formyl-5-(5-(hydroxymethyl)thiophen-3-yl)pyrazolo[1,5-a]pyrimidine-7-yl)carbamate (638 mg, 69%) as an off white solid. ¹H NMR (CDCl₃, 400 MHz) δ: 10.34 (s, 1H, 8.55 (s, 1H), 8.11 (d, 1H, J = 1.6 Hz), 7.76 (d, 1H, J = 1.6 Hz), 7.18 (s, 1H), 4.93 (bs, 2H), 3.30 (dddd, 1H, J = 6.8, 6.8, 3.6, 3.6 Hz), 2.15 (bs, 1H), 1.42 (s, 9H), 0.85-0.92 (m, 2H), 0.63-0.70 (m, 2H). LCMS (ES): >95% pure, m/z 415 [M+1]⁺.

Tert-butyl cyclopropyl(3-formyl-5-(5-(hydroxymethyl)thiophen-3-yl)pyrazolo[1,5-a]pyrimidine-7-yl)carbamate (20 mg, 0.05 mmol) was dissolved in dichloromethane (0.5 mL) and trifluoroacetic acid (0.5 mL). After 1 h, the solution was concentrated under a stream of air. The residue was purified via preparative HPLC to furnish 7-(cyclopropylamino)-5-(5-(hydroxymethyl)thiophen-3-yl)pyrazolo[1,5-a]pyrimidine-3-carbaldehyde 2,2,2-trifluoroacetate (4.8 mg, 23%).

Hydrogen bromide (48% in water, 2.5 mL) was added dropwise to tert-butyl cyclopropyl(3-formyl-5-(5-(hydroxymethyl)thiophen-3-yl)pyrazolo[1,5-a]pyrimidine-7-yl)carbamate (561 mg, 1.35 mmol) suspended in dichloromethane (3.5 mL). The solution immediately became dark brown and homogeneous upon addition. The reaction was heated to 40°C for 3 h, then diluted with dichloromethane (10 mL). The liquid was decanted and the gummy residue was washed with dichloromethane (3 x 10 mL). The combined liquids were washed successively with sat. NaHCO₃ (20 mL) and brine (20 mL), and then dried over MgSO₄ filtered and concentrated in vacuo. The residue was triturated with hexanes and then purified via flash column chromatography (15-40% EtOAc/hexanes) to provide 5-(5-(bromomethyl)thiophen-3-yl)-7-(cyclopropylamino)pyrazolo[1,5-a]pyrimidine-3-carbaldehyde (105 mg, 20%) as a yellow solid. ¹H NMR (CDCl₃, 400 MHz) δ: 10.26 (s, 1H), 8.44 (s, 1H), 8.12 (d, 1H, J = 1.6 Hz), 7.80 (s, 1H), 6.73 (s, 1H), 6.65 (bs, 1H), 4.80 (s, 2H), 2.81 (m, 1H), 1.03-1.09 (m, 2H), 0.84-0.89 (m, 2H). LCMS (ES): >95% pure, m/z 378 [M+1]⁺.

Potassium carbonate (30 mg, 0.20 mmol) was added to 5-(5-(bromomethyl)thiophen-3-yl)-7-(cyclopropylamino)pyrazolo[1,5-a]pyrimidine-3-carbaldehyde (25 mg, 0.07 mmol) dissolved in DMF (0.7 mL). Pyrrolidine (6 µL, 0.07 mmol) was added and the reaction was heated to 50°C for 1.25 h. Water (3 mL) was added and the orange precipitate was filtered and dried in vacuo to give 7-(cyclopropylamino)-5-(5-(pyrrolidin-1-ylmethyl)thiophen-3-yl)pyrazolo[1,5-a]pyrimidine-3-carbaldehyde (13 mg, 54%) which was used without further purification. LCMS (ES): >85% pure, m/z 368 [M+1]⁺.

Hydantoin (3 mg, 0.03 mmol) and piperidine (3 µL, 0.03 mmol) were added to 7-(cyclopropylamino)-5-(5-(pyrrolidin-1-ylmethyl)thiophen-3-yl)pyrazolo[1,5-a]pyrimidine-3-carbaldehyde (12 mg, 0.03 mmol) dissolved in ethanol (0.5 mL). The reaction was heated at 80 °C. After 15 h, the reaction was cooled to r.t., then diluted with water (3 mL). The precipitate was collected and washed with 1:1 ethanol:water (3 mL) and dried in vacuo to furnish (Z)-5-((7-(cyclopropylamino)-5-(5-pyrrolidin-1-ylmethyl)thiophen-3-yl)pyrazolo[1,5-a]pyrimidin-3-yl)methylene)imidazolidine-2,4-dione (2.8 mg, 9% over two steps). LCMS (ES): >95% pure, m/z 450 (M+1]⁺.

The compounds described in the following table were prepared using chemistries similar to those exemplified in Example 174. All compounds were characterized by LCMS. Table 26B shows the biological activities of the compounds listed in Table 26A. Compound CK2: IC50 (µM) PIM2: IC50 (5 µM ATP) AB: MDAMB453 (µM) AB: BxPC3 (µM) Y13 <0.01 > 2.5000 1.386 0.929 Z13 <0.01 > 2.5000 2.498 10.153 A14 <0.01 1.5722 1.614 1.758 B14 <0.01 1.4451 1.614 1.003

The chemistry depicted in Figure 3 can be used to prepare analogs 7 substituted by a methyl group. Commercially available boronic acid 1 can be reacted with tert-butyl 5-chloropyrazolo[1,5-a]pyrimidin-7-yl(cyclopropyl)carbamate 2 under Suzuki reaction conditions to for methyl ketone 3. This compound 3 can be reacted with various substituted amines 4 under reductive amination conditions such as the conditions described in US2007/244094 or reaction conditions described in European Journal of Medicinal Chemistry, vol 32, 1997, 143-150 to prepare compounds 5. Compound 5 can be converted to aldehyde 6 under Vilsmeier conditions. Compound 6 can be converted to compound 7 by reacting with hydantoin and piperidine in ethanol.

The molecules in the following Table 27 can be prepared using similar chemistries.

Diisopropylethylamine (2.4 mL, 13.62 mmol) and cyclopropylamine (943 µL, 13.62 mmol) were added to commercially available (Ark Pharm, Inc.) 6,8-dibromoimidazo[1,2-a]pyrazine (2.51 g, 9.08 mmol) dissolved in 2-propanol (9 mL). The solution was placed in an 80 °C oil bath. After 4.5 h, the volatiles were removed in vacuo. The brown residue was partitioned between dichloromethane (50 mL) and water (50 mL). The organic layer was washed further with water (50 mL) and then brine (50 mL). The organic layer was dried over MgSO₄, filtered and concentrated in vacuo. The residue was purified via a filtration over a short plug of silica gel (40% EtOAc/hexanes) and the filtrate was concentrated in vacuo to afford 6-bromo-N-cyclopropylimidazo[1,2-a]pyrazin-8-amine (2.19 g, 95%) as a light brown solid. ¹H NMR (CDCl₃, 400 MHz) δ: 7.61 (s, 1H), 7.46 (d, 1H, J = 1.2 Hz), 7.44 (d, 1H, J = 1.2 Hz), 6.26 (bs, 1H), 3.02 (dddd, 1H, J = 7.2, 7.2, 7.2, 3.6 Hz), 0.89-0.95 (m, 2H), 0.64-0.69 (m, 2H). LCMS (ES): >95% pure, m/z 254 [M+1]⁺.

6-Bromo-N-cyclopropylimidazo[1,2-a]pyrazin-8-amine (0.5 g, 1.98 mmol) was dissolved in dichloromethane (8 mL). Di-tert-butyl dicarbonate (733 mg, 3.35 mmol), DMAP (5 mg, 0.02 mmol) and pyridine (0.4 mL) were added sequentially. After 12 h, the solution was diluted with EtOAc (50 mL) and then washed sequentially with 1N HCl (50 mL), 1N NaOH (50 mL), and brine (50 mL). The organic layer was dried over MgSO₄ filtered and concentrated in vacuo. The residue was triturated with hexanes (5 mL) to yield tert-butyl 6-bromoimidazo[1,2-a]pyrazin-8-yl(cyclopropyl) carbamate (337 mg, 48%) as an off white solid. ¹H NMR (CDCl₃, 400 MHz) δ: 8.18 (s, 1H), 7.78 (d, 1H, J = 0.8 Hz), 7.67 (d, 1H, J = 0.8 Hz), 3.25 (dddd, 1H, J = 6.8, 6.8, 3.6, 3.6 Hz), 1.20 (s, 9H), 0.78-0.86 (m, 2H), 0.71-0.77 (m, 2H). LCMS (ES): >90% pure, m/z 354 [M+1]⁺.

Phosphorus(V) oxychloride (3.9 mL, 42.68 mmol) was added dropwise to anhydrous DMF (16 mL) at 0 °C. 6-bromo-N-cyclopropylimidazo[1,2-a]pyrazin-8-amine (900 mg, 3.56 mmol) was dissolved in anhydrous DMF (24 mL) and added over two minutes. The solution was place in an 85°C oil bath for 5 h. The solution was cooled to 0°C and conc. HCl (30 mL) was added. The mixture was basified to pH=10w/ 3N NaOH (∼175 mL). The mixture was extracted with dichloromethane (3 x 250 mL), and the organics were washed with brine (500 mL). The organic layer was dried over MgSO₄, filtered and concentrated in vacuo. The residue was purified via flash column chromatography (30% EtOAc/hexanes) to furnish 6-bromo-8-(cyclopropylamino)imidazo[1,2-a]pyrazine-3-carbaldehyde (490 mg, 49%). LCMS (ES): >95% pure, m/z 282 [M+H]⁺.

Di-tert-butyl dicarbonate (1.16 g, 5.30 mmol) and DMAP (21 mg, 0.18 mmol) were added to a solution of 6-bromo-8-(cyclopropylamino)imidazo[1,2-a]pyrazine-3-carbaldehyde (994 mg, 3.50 mmol) in dichloromethane (15 mL). After 2.5 h, the solution was partitioned between EtOAc (100 mL) and water (100 mL). The aqueous layer was further extracted with EtOAc (2 x 75 mL). The organics were washed with brine (250 mL), dried over MgSO₄ filtered and concentrated in vacuo. The residue was purified via flash column chromatography (30% EtOAc/hexanes) to provide tert-butyl 6-bromo-3-formylimidazo[1,2-a]pyrazine-8-yl(cyclopropyl)carbamate (1.17 g, 87%) as a brown foam. ¹H NMR (CDCl₃, 400 MHz) δ: 10.05 (s, 1H), 9.42 (s, 1H), 8.37 (s, 1H), 3.25 (dddd, 1H, J = 6.8, 6.8, 4.0, 4.0 Hz), 1.22 (s, 9H), 0.85-0.90 (m, 2H), 0.69-0.75 (m, 2H). LCMS (ES): >95% pure, m/z 382 [M+1]⁺.

Tert-butyl 6-bromo-3-formylimidazo[1,2-a]pyrazine-8-yl(cyclopropyl)carbamate (130 mg, 0.34 mmol), 3-(trifluoromethoxy)phenyl boronic acid (105 mg, 0.51 mmol), 3M Na₂CO₃ (1.1 mL, 3.4 mmol) and DME (4.5 mL) were combined. The solution was degassed with a stream of N₂ for 10 min. Pd(PPh₃)₄ was added and the solution was refluxed for 2 h. The solution was partitioned between dichloromethane (25 mL) and water (25 mL). The aqueous layer was further extracted with dichloromethane (2 x 25 mL). The organics were washed with brine (50 mL), dried over MgSO₄, filtered and concentrated in vacuo. The residue was purified via flash column chromatography (30-45% EtOAc/hexanes) to provide tert-butyl cyclopropyl(3-formyl-6-(3-trifluoromethoxy)phenyl)imidazo[1,2-a]pyrazin-8-yl)carbamate (96 mg, 61%) as a bright yellow solid. LCMS (ES): >95% pure, m/z 463 [M+1]⁺.

Tert-butyl cyclopropyl(6-(3-trifluoromethoxy)phenyl)imidazo[1,2-a]pyrazin-8-yl)carbamate (77%) was synthesized in a manner analogous to Example 179. LCMS (ES): >95% pure, m/z 435 [M+1]⁺.

Tert-butyl cyclopropyl(6-(3-fluorophenyl)-3-formylimidazo[1,2-a]pyrazin-8-yl)carbamate (28%) was synthesized in a manner analogous to Example 183. LCMS (ES): >95% pure, m/z 435 [M+1]⁺.

Tert-butyl 6-bromo-3-formylimidazo[1,2-a]pyrazine-8-yl(cyclopropyl)carbamate (100 mg, 0.26 mmol), 4-[3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzyl]morpholine (118 mg, 0.39 mmol), 3M Na₂CO₃ (1.3 mL, 2.60 mmol) and DME (3.5 mL) were combined. The solution was degassed with a stream of N₃ for 10 min. Pd(PPh₃)₄ was added and the solution was refluxed for 2 h. The solution was partitioned between dichloromethane (25 mL) and water (25 mL). The aqueous layer was further extracted with dichloromethane (2 x 25 mL). The organics were washed with brine (50 mL), dried over MgSO₄, filtered and concentrated in vacuo. The residue was purified via preparative TLC (5% MeOH/dichloromethane) to afford tert-butyl cyclopropyl(3-formyl-6-(3-(morpholinomethyl)phenyl)imidazo[1,2-a]pyrazin-8-yl)carbamate (60 mg, 48%). LCMS (ES): >95% pure, m/z 478 [M+1]⁺

Triethylamine (9.12 µL, 6.56 mmol) was added to tert-butyl 6-bromo-3-formylimidazo[1,2-a]pyrazine-8-yl(cyclopropyl)carbamate (250 mg, 0.66 mmol) dissolved in anhydrous DMF (2.2 mL) in a 15 mL pressure tube. The solution was degassed with a stream of N₂ for 10 min. Trimethylsilylacetylene (927 µL, 6.56 mmol), Pd(PPh₃)₄ (76 mg, 0.07 mmol), and copper(I) iodide (25 mg, 0.13 mmol) were added and the reaction was sealed and heated to 65 °C for 24 h. The reaction was diluted with EtOAc (50 mL) and then washed with 10% brine (4 x 50 mL) and brine (50 mL). The organics were dried over MgSO₄, filtered and concentrated in vacuo. The residue was purified via flash column chromatography (30% EtOAc/hexanes) to give tert-butyl cyclopropyl(3-formyl-6-((trimethylsilyl)ethynyl)imidazo[1,2-a]pyrazin-8-yl)carbamate (186 mg, 71%) as a brown foamy solid. LCMS (ES): >95% pure, m/z 400 [M+1]⁺.

Tert-butyl cyclopropyl(3-formyl-6-((phenylethynyl)imidazo[1,2-a]pyrazin-8-y])carbamate (64%) was synthesized in a manner analogous to Example 183. LCMS (ES): >95% pure, m/z 403 [M+1]⁺.

Potassium carbonate (86 mg, 0.63 mmol) was added to tert-butyl cyclopropyl(3-formyl-6-((trimethylsilyl)ethynyl)imidazo[1,2-a]pyrazin-8-yl)carbamate (50 mg, 0.13 mmol) dissolved in methanol (2.5 mL). After 2 h, the volatiles were removed in vacuo. The residue was partitioned between dichloromethane (10 mL) and water (10 mL). The aqueous layer was further extracted with dichloromethane, (2 x 10 mL). The organics were washed with brine (30 mL), dried over MgSO₄, filtered and concentrated in vacuo. The residue was purified via flash column chromatography (30% EtOAc/hexanes) to provide tert-butyl cyclopropyl(6-ethynyl-3-formylimidazo[1,2-a]pyrazin-8-yl)carbamate (20 mg, 50%) as a yellow foamy solid. LCMS (ES): >95% pure, m/z 327 [M+1]⁺.

Hydantoin (33 mg, 0.33 mmol) and piperidine (33 µL, 0.33 mmol) were added to tert-butyl cyclopropyl(3-formyl-6-(3-trifluoromethoxy)phenyl)imidazo[1,2-a]pyrazin-8-yl)carbamate (50 mg, 0.11 mmol) suspended in ethanol (0.5 mL). The reaction was sealed and irradiated in the microwave at 80 °C for 12 h. The precipitate was filtered off and washed with ethanol (3 mL) to give (Z)-tert-butyl cyclopropyl(3-((2,5-dioxoimidazolidin-4-ylidene)methyl)-6-(3-trifluoromethoxy)phenyl)imidazo[1,2-a]pyrazin-8-yl)carbamate (18 mg, 30%) as a bright yellow solid. LCMS (ES): >90% pure, m/z 545 [M+1]⁺.

Tert-butyl cyclopropyl(3-((2,5-dioxoimidazolidin-4-ylidene)methyl)-6-(3-trifluoromethoxy)phenyl)imidazo[1,2-a]pyrazin-8-yl)carbamate (15 mg, 0.03 mmol) was dissolved in dichloromethane (0.5 mL) and trifluoroacetic acid (0.5 mL). After 1 h, the solution was concentrated under a stream of air. The residue was purified via preparative HPLC to furnish (Z)-5-((8-cyclopropylamino)-6-(3-(trifluoromethoxy)phenyl) imidazo[1,2-a]pyrazin-3-yl)methylene)imidazolidine-2,4-dione (0.9 mg, 8%).

Hydantoin (24 mg, 0.24 mmol) and piperidine (24 µL, 0.24 mmol) were added to tert-butyl cyclopropyl(3-formyl-6-((phenylethynyl)imidazo[1,2-a]pyrazin-8-yl)carbamate (24 mg, 0.06 mmol) dissolved in ethanol (1 mL). The reaction was heated at 80°C for 12 h, and then cooled to r.t. The precipitate was filtered off and washed with ethanol (3 mL) to give (Z)-tert-butyl cyclopropyl(3-((2,5-dioxoimidazolidin-4-ylidene)methyl)-6-(phenylethynyl)imidazo[1,2-a]pyrazin-8-yl)carbamate (12 mg, 43%) as an orange/yellow solid. LCMS (ES): >90% pure, m/z 485 [M+1]⁺.

(Z)-Tert-butyl cyclopropyl(3-((2,5-dioxoimidazolidin-4-ylidene)methyl)-6-(phenylethynyl)imidazo[1,2-a]pyrazin-8-yl)carbamate (12 mg, 0.03 mmol) was dissolved in dichloromethane, (0.3 mL) and trifluoroacetic acid (0.3 mL). After 1 h, the solution was concentrated under a stream of air. The residue was triturated with Et₂O and filtered to yield (Z)-5-((8-(cyclopropylamino)-6-(phenylethynyl)imidazo[1,2-a]pyrazin-3-yl)methylene)imidazolidine-2,4-dione (6 mg, 63%) as a bright yellow solid.

Hydantoin (152 mg, 1.50 mmol) and piperidine (150 µL, 1.50 mmol) were added to tert-butyl cyclopropyl(3-formyl-6-(3-(morpholinomethyl)phenyl)imidazo[1,2-a]pyrazin-8-yl)carbamate (60 mg, 0.13 mmol) dissolved in ethanol (1 mL). The reaction was heated at 80°C for 4 d, and then diluted with water (10 mL). The supernatant was decanted and extracted with dichloromethane (2 x 5 mL). The organics were washed with brine (30 mL), dried over MgSO₄, filtered and concentrated in vacuo to a yellow solid. LCMS (ES): >95% pure, m/z 560 [M+1]⁺.

The crude solid was dissolved in dichloromethane (0.5 mL) and trifluoroacetic acid (0.5 mL). After 1 h, the solution was concentrated under a stream of air. The residue was purified via preparative HPLC to furnish (Z)-5-((8-(cyclopropylamino)-6-(3-morpholinomethyl)phenyl)imidazo[1,2-a]pyrazin-3-yl)methylene)imidazolidine-2,4-dione 2,2,2-trifluoroacetate (5.5 mg, 8% over two steps).

Hydantoin (18 mg, 0.17 mmol) and piperidine (17 µL, 0.17 mmol) were added to tert-butyl cyclopropyl(6-fluorophenyl)-3-formyllimidazo[1,2-a]pyrazin-8-yl)carbamate (23 mg, 0.06 mmol) dissolved in ethanol (0.3 mL). The reaction was heated at 80°C for 18 h, and then concentrated in vacuo to a yellow solid. The crude solid was dissolved in dichloromethane, (0.5 mL) and trifluoroacetic acid (1.5 mL). After 1 h, the solution was concentrated under a stream of air. The residue was triturated with ethanol and filtered to provide (Z)-5-((8-cyclopropylamino)-6-(3-fluorophenyl)imidazo[1,2-a]pyrazin-3-yl)methylene)imidazolidine-2,4-dione as an orange/yellow solid (2.4 mg, 10% over two steps).

The compounds in the following table were prepared by the methods described above, by selecting appropriate starting materials as is apparent to the person of ordinary skill. Table 28B shows the biological activities of the compounds listed in Table 28A. Compound CK2: IC50 (µM) PIM2: IC50 (5 µM ATP) AB: MDAMB453 (µM) AB: BxPC3 (µM) C14 > 5.0000 > 2.5000 D14 <0.1 > 2.5000 4.531 3.69 E14 > 5.0000 > 2.5000 F14 > 5.0000 > 2.5000 G14 > 5.0000 > 2.5000

The chemistries described on Figure 7 can be used to prepare analogs substituted by a trifluoromethyl group. Commercially available 2-amino-3,5-dibromopyrazine and commercially available 3-bromo-1,1,1-trifluoroacetone can be reacted together at 50°C in a solvent such as dioxin (conditions previously described in WO2003/82817), to prepare compound 3 Compound 3 can be reacted with amine R₁NH₂ to obtain 4. This material can be protected by a boc group by reacting 4 with a reagent like Boc₂O to obtain 5. This material can be further transformed into 6 under vilsmeir conditions in the presence of POCl₃. Compound 6 can be reacted with various reagents such as boronic acids or esters W-B(OR₃)₂ under Suzuki conditions to form molecule 7.

Other analogs of 7 can be prepared by heating 6 with amines or anilines R₅R₆NH, alcohols or phenols R₅OH, thiols or thiophenols R₅SH, in the presence of a base or an acid. Compound 8 can be prepared by heating 7 with hydantoin in ethanol in the presence of a base such as piperidine.

Unless otherwise specified, the various substituents of the compounds are defined in the same manner as the formulae II/II' compound of the invention.

The chemistry described in Figure 4 and Figure 5 can be used to prepare various substituted compounds of formula II.

Substituted aminopyrazole 1 can react with isothiocyanate 2 to form intermediate 3. Compound 3 can be cyclized to 4 in the presence of a base such as sodium hydroxide. Compound 4 can be alkylated by with R7Halo in the presence of a base. Compound 5 can be converted to compound 6 using phosphorus oxychloride. Molecule 7 can be prepared by addition of amine R₇R₈NH to molecule 6 in a solvent like NMP or DMF. Compound 8 can be obtained by reacting compound 7 with DMF and Phosphorus oxychloride under Vilsmeier reaction conditions. Aldehyde 8 can be converted in two steps to substituted ketone 8b by reacting with a Grignard reagent R₄MgX, followed by reaction with an oxidant such as DCC or using Swern reaction conditions.

Compound 8 and 8a, or 8b and 8a can react upon heating in a solvent such as ethanol and in the presence of a base such as piperidine to form compound 9. Oxidation of 9 by an oxidant such as meta-chloroperbenzoic acid or oxone can provide compound 10, which can contain variable quantities of sulfide (n = 0), sulfoxide (n = 1) or sulfone (n = 2).

The chemistry depicted in Figure 5 can be used to prepare various substituted analogs of formula II compounds.

Compound 10 can be mixed at room temperature or heated with amines R₇R₈NH to form compound 11. Compound 10 can be reacted with hydrazines R₇R₈N-NH₂ to form compound 12. Compound 10 can be reacted with alcohols or phenols R₇OH in the presence of a base such as NaH or K₂CO₃ to form compound 13. Compound 10 can be reacted with thiols or thiophenols R₇SH with or without a base to form compound 14.

The chemistry described in Figure 6 can be used to prepare analogs substituted by aryl or heteroaryls groups. Compound 7 can be reacted with boronic esters or acids W-B(OR⁷)₂ or organo tin compounds W-Sn(R⁷)₃ in the presence of tri(2-furyl)phosphine, copper(I)thiophene-2-carboxylate and Pd₂dba₃ or using conditions previously desbribed in Organic Letters 2002, vol 4(6), pp. 979-981. Compound 15 can be converted to compound 18 using chemistries similar to the one described in Figure 4.

The material was prepared according to a procedure published in patent US 3,846,423. Characterized by LCMS (ES):>95% pure, m/z 183 [M+H]⁺.

In a round bottom flask equipped with a magnetic stirbar, 2-methylthio)pyrazolo[1,5-a][1,3,5]triazin-4(3H)-one (1.0 eq, 10.43 g, 57.24 mmol) was suspended in acetonitrile (100 ml). Phosphorus oxychloride (4.0 eq, 21 ml, 229.4 mmol) and triethylamine (1.05 eq, 8.4 ml, 60.27 mmol) were added and the mixture stirred at reflux for 3.5 hours, at which time LCMS indicated completion of the reaction. The mixture was cooled down and slowly poured into crushed ice (final total volume of about 600 ml). The solid was filtered, washed with water and dried in a vacuum oven to afford 4-chloro-2-(methylthio)pyrazolo[1,5-a][1,3,5]triazine as a tan solid (8.15 g, 71% yield). LCMS (ES):>97% pure, m/z 201 [M+1]⁺.

4-Chloro-2-(methylthio)pyrazolo[1,5-a][1,3,5]triazine (1.0 eq, 6.26 g, 31.19 mmol) was suspended in anhydrous NMP (50 ml). Cyclopropylamine (1.5 eq, 3.2 ml, 46.26 mmol) was added through syringe dropwise. Internal temperature rose to 47°C. The mixture was stirred without any external cooling for one hour. An additional amount of cypropylamine (1ml) was added and the mixture stirred for another 1.5 hours. The mixture was slowly poured into water (500 ml) under stirring. The resulting solid was filtered, washed with water and dried in a vacuum oven to give N-cyclopropyl-2-(methylthio)pyrazolo[1,5-a][1,3,5]triazin-4-amine as a tan solid (5.44 g, 79% yield). LCMS (S):>95% pure, m/z 222 [M+H]⁺.

The following molecules were prepared using chemistries similar to Example 195. Compounds were characterized by LCMS. Structure MW LCMS m/z |M+1]+ 253.32 254 271.34 272 263.24 264 235.31 236

N-Cyclopropyl-2-(methylthio)pyrazolo[1,5-a][1,3,5]triazin-4-amine (1.0 eq, 3.10 g, 14.00 mmol) was dissolved in anhydrous DMF (50 ml) under nitrogen atmosphere. Phosphorus oxychloride (5.0 eq, 6.4 ml, 69.9 mmol) was added dropwise over 5 minutes. Internal temperature rose to 45°C. The reaction was stirred in an oil bath at 70°C for 4.5 hours. The mixture was cooled down and added dropwise into a solution of 6N NaOH (150 ml) chilled with an ice bath. The rate of addition was adjusted to maintain the internal temperature of the aqueous NaOH below 16°C. At the end of the addition, the mixture was neutralized by slow addition of 6N HCl to reach pH = 5-6. The resulting solid was filtered, washed with water and dried in a vacuum oven overnight. 4-(cyclopropylamino)-2-(methylthio)pyrazolo[1,5-a][1,3,5]tnazine-8-carbaldehyde was isolated as tan solid (9.26 g, 93%). LCMS (ES):>95% pure, m/z 250 [M+H]⁺.

The following molecules were prepared using chemistries similar to Example 196. Compounds were characterized by LCMS. Structure MW LCMS m/z [M+1]+ 281.3341 282 299.3509 300 291.26 292 263.32 264

4-(Cyclopropylamino)-2-(methylthio)pyrazolo[1,5-a][1,3,5]triazine-8-carbaldehyde (1.0 eq, 3.00 g, 12.03 mmol) was suspended in ethanol (40 ml). Hydantoin (1.5 eq, 1.81 g, 18.08 mmol) and piperidine (1.5 eq, 1.78 ml, 18.01 mmol) were added. The mixture was heated at reflux under vigorous magnetic stirring for 3 hours. After cooling of the reaction mixture, the precipitate was filtered, washed with ethanol, then with a mixture of ethanol and water (1:1). After drying in vacuo, (Z)-5-((4-(cyclopropylamino)-2-(methylthio)pyrazolo[1,5-a][1,3,5]triazin-8-yl)methylene)imidazolidine-2,4-dione was isolated as a yellow solid (3.80 g, 95%). LCMS (ES):>85% pure, m/z 332 [M+H]⁺.

The following molecules were prepared using chemistries similar to Example 197. Compounds were characterized by LCMS. Structure MW LCMS m/z [M+1]+ 363.39 364 373.32 374 381.42 382

(Z)-5-((4-(Cyclopropylamino)-2-(methythio)pyrazolo[1,5-a][1,3,5]triazin-8-yl)methylene)imidazolidine-2,4-dione (1.0 eq, 3.00 g, 9.05 mmol) was suspended in dichloromethane (150 ml). m-cpba (77% purity grade, 5.0 eq, 10.1 g, 45.06 mmol) was added and the mixture stirred at room temperature for 4 hours. The reaction was diluted by addition of dichloromethane (500 ml). The solid was filtered and washed with dichloromethane. After drying a (1:1) mixture of (Z)-5-((4-(cyclopropylamino)-2-(methylsulfonyl)pyrazolo[1,5-a][1,3,5]triazin-8-yl)methylene)imidazolidine-2,4-dione and (Z)-5-((4-(cyclopropylamino)-2-(methylsulfinyl)pyrazolo[1,5-a][1,3,5]triazin-8-yl)methylene)imidazolidine-2,4-dione was isolated as a yellow solid (2.67 g, 81%). LCMS (ES):>85% pure, m/z 364 [M+H]⁺ and m/z 398 [M+H]⁺. The mixture was used for next step without any separation of the molecules.

The following mixtures of sulfones and sulfoxides were prepared using chemistries similar to Example 198. Compounds were characterized by LCMS. Structure MW LCMS m/z [M+1]⁺ Structure MW LCMS m/z [M+1]+ 405.32 406 389.32 390 395.4 396 379.4 380

A (1:1) mixture of (Z)-5-((4-(cyclopropylamino)-2-(methylsulfonyl)pyrazolo[1,5-a][1,3,5]triazin-8-yl)methylene)imidazolidine-2,4-dione and (Z)-5-((4-(cyclopropylamino)-2-(methylsulfinyl)pyrazolo[1,5-a][1,3,5]triazin-8-yl)methylene)imidazolidine-2,4-dione (15 mg) was mixed with 3-chloroaniline (0.1 ml) in NMP (0.2 ml) and the mixture heated in a microwave oven at 120°C for 15 min. Methanol was added and the resulting solid filtered and dried to provide (Z)-5-((2-(3-chlorophenylamino)-4-(cyclopropylamino)pyrazolo[1,5-a][1,3,5]triazin-8-yl)methylene)imidazolidine-2,4-dione as a solid (7 mg). LCMS (E):>95% pure, m/z 411 [M+H]⁺.

A (1:1) mixture of (Z)-5-((4-(cyclopropylamino)-2-(methylsulfonyl)pyrazolo[1,5-a][1,3,5]triazin-8-yl)methylene)imidazolidine-2,4-dione and (Z)-5-((4-(cyclopropylamino)-2-(methylsulfinyl)pyrazolo[1,5-a][1,3,5]triazin-8-yl)methylene)imidazolidine-2,4-dione (36 mg) was suspended in NMP (0.2 ml). Cyclopropylmethylamine (88 uL) was added and the mixture stirred at room temperature for 15 minutes. Water and methylene chloride were added and the resulting precipitate was filtered. After triturating in a mixture of ethyl acetate and hexanes, (Z)-5-((4-(cyclopropylamino)-2-(cyclopropylmethylamino)pyrazolo[1,5-a][1,3,5]triazin-8-yl) methylene) imidazolidine-2,4-dione was isolated as a yellow solid.). LCMS (ES):>95% pure, m/z 355 [M+H]⁺.

A (1:1) mixture of (Z)-5-((4-(cyclopropylamino)-2-(methylsulfonyl)pyrazolo[1,5-a][1,3,5]triazin-8-yl)methylene)imidazolidine-2,4-dione and (Z)-5-((4-(cyclopropylamino)-2-(methylsulfinyl)pyrazolo[1,5-a][1,3,5]triazin-8-yl)methylene)imidazolidine-2,4-dione (1.0 eq, 25 mg, 0.0704 mmol) was combined in a vial with 3-chlorophenol (5.0 eq, 45 mg, 0.35 mmol) and K₂CO₃ (5.0 eq, 48 mg, 0.347 mmol) in NMP (0.2 ml). The mixture was stirred at 90°C for 1 hour. Water was added and the resulting solid was filtered and dried. Trituration in a mixture of ethyl acetate and hexanes followed by filtration provided (Z)-5-((2-(3-chlorophenoxy)-4-(cyclopropylamino)pyrazolo[1,5-a][1,3,5]triazin-8-yl)methylene)imidazolidine-2,4-dione as a tan solid (20 mg, 69%). LCMS (E):>95% pure, m/z 412 [M+H]⁺.

(1r,4r)-4-(4-(Cyclopropylamino)-8-((Z)-(2,5-dioxoimidazolidin-4-ylidene)methyl)pyrazolo[1,5-a][1,3,5]triazin-2-ylamino)cyclohexanecarboxylic acid (1 eq, 12 mg, 0.028 mmol) was mixed in NMP (0.4 ml) with methyl amine hydrochloride (8 eq, 15 mg, 0.225 mmol), HOBt.H₂O (2 eq, 8 mg, 0.056 mmol), DIEA (4 eq, 14 uL, 0.113 mmol) and EDCI (4 eq, 22 mg, 0.113 mmol). The mixture was stirred at 70°C for 2.5 hours. Water was added and the precipitate filtered to afford (1r,4r)-4-(4-(cyclopropylamino)-8-((Z)-(2,5-dioxoimidazolidin-4-ylidene)methyl)pyrazolo[1,5-a][1,3,5]triazin-2-ylamino)-N-methylcyclohexanecarboxamide. LCMS (E):>95% pure, m/z 440 [M+H]⁺.

(1r,4r)-4-(4-(Cyclopropylamino)-8-((Z)-(2,5-dioxoimidazolidin-4-ylidene)methyl)pyrazolo[1,5-a][1,3,5]triazin-2-ylamino)cyclohexanecarboxylic acid (1 eq, 12 mg, 0.028 mmol) was mixed in NMP (0.4 ml) with ammonium chloride (8 eq, 12 mg, 0.225 mmol), HOBt.H₂O (2 eq, 8 mg, 0.056 mmol), DIEA (4 eq, 14 uL, 0.113 mmol) and EDCI (4 eq, 22 mg, 0.113 mmol). The mixture was stirred at 70°C for 2.5 hours. Water was added and the precipitate filtered to afford (1r,4r)-4-(4-(cyclopropylamino)-8-((Z)-(2,5-dioxoimidazolidin-4-ylidene)methyl)pyrazolo[1,5-a][1,3,5]triazin-2-ylamino)cyclohexanocarboxamide. LCMS (ES):>95% pure, m/z 426 [M+H]⁺.

A (1:1) mixture of (Z)-5-((4-(cyclopropylamino)-2-(methylsulfonyl)pyrazolo[1,5-a][1,3,5]triazin-8-yl)methylene)imidazolidine-2,4-dione and (Z)-5-((4-(cyclopropylamino)-2-(methy)sulfinyl)pyrazolo[1,5-a][1,3,5]triazin-8-yl)methylene)imidazolidine-2,4-dione (1.0 eq, 16 mg, 0.0451 mmol) was reacted with trans-1,4-diaminocyclohexane (20.0 eq, 103 mg, 0.902 mmol) in NMP (0.4 ml) at room temperature for 3 hours. Water and methanol was added and the material was purified by preparative HPLC. Genevac evaporation provided (Z)-5-((2-((1r,4r)-4-aniinocyclohoxylamino)-4-(cyclopropylamino)pyrazolo[1,5-a][1,3,5]triazin-8-yl)methylene)imidazolidine-2,4-dione 2,2,2-trifluoroacetate (15 mg). LCMS (ES):>95% pure, m/z 398 [M+H]⁺.

Di-tert-butyl dicarbonate (327 mg, 1.50 mmol) and DMAP (6 mg, 0.05 mmol) were added to N-cyclopropyl2-(methylthio)pyrazolo[1,5-a][1,3,5]triazin-4-amine (221 mg, 1 mmol) dissolved in dichloromethane (4 mL). After 15 h, the solution was diluted with EtOAc (100 mL) and washed successively with water (3 x 100 mL) and brine (100 mL). The organic layer was dried over MgSO₄, filtered and concentrated in vacuo to an orange oil. The residue was purified via flash column chromatography (10% EtOAc/hexanes) to afford tert-butyl cyclopropyl(2-(methylthio)pyrazolo[1,5-a][1,3,5]triazin-4-yl)carbamate (368 mg, 79%). LCMS (ES): >95% pure, m/z 322 [M+1]⁺.

Note: THF was degassed with a stream of N₂ for 10 min. in a separate flask. Tert-butyl cyclopropyl(2-(methylthio)pyrazolo[1,5-a][1,3,5]triazin-4-yl)carbamate (100 mg, 0.31 mmol), 3-(trifluoromethoxy)phenyl boronic acid (154 mg, 0.74 mmol), tri(2-furyl)phosphine (86 mg, 0.37 mmol), copper(I) thiophene-2-carboxylate (167 mg, 0.88 mmol), Pd₂dba₃ (24 mg, 0.03 mmol) were combined. The flask was evacuated and backfilled with N₂. THF (3.7 mL) was added and the reaction was heated to 50°C for 5 d. The solution was diluted with Et₂O (40 mL) and washed with 10% NH₄OH (3 x 30 mL). The organic layer was dried over MgSO₄, filtered and concentrated in vacuo. The solid residue was triturated with H₂O and filtered. The filtrate was concentrated in vacuo and purified via flash column chromatography (2.5-5% EtOAc/hexanes) to afford tert-butyl cyclopropyl(2-(3-(trifluoromethoxy)phenyl)pyrazolo[1,5-a][1,3,5]triazin-4-yl)carbamate (116 mg, 85%). LCMS (ES): >95% pure, m/z 436 [M+1]⁺.

Tert-butyl cyclopropyl(2-(3-(trifluoromethoxy)phenyl)pyrazolo[1,5-a][1,3,5]triazin-4-yl)carbamate was dissolved in dichloromethane (0.7 mL) and trifluoroacetic acid (0.7 mL). After 1 h, the solution was concentrated under a stream of air to give crude 4-(cyclopropylamino)-2-(3-(trifluoromethoxy)phenyl)pyrazolo[1,5-a][1,3,5]triazine-8-carbaldehyde which was used without further purification. LCMS (ES): >90% pure, m/z 336 [M+1]⁺. 4-(cyclopropylamino)-2-(3-(trifluoromethoxy)phenyl)pyrazolo[1,5-a][1,3,5]triazine-8-carbaldehyde (87 mg, 0.26 mmol) was dissolved in DMF (0.8 mL). Phosphorus(V) oxychloride (318 µL, 3.47 mmol) was added was added dropwise and the reaction was heated to 70 °C. After 6 h, the solution was added dropwise to 6M NaOH (∼10 mL) cooled to 0 °C. The pH was adjusted to 7 by the addition of 12N HCl. The precipitate was filtered off and dried in vacuo to furnish 4-(cyclopropylamino)-2-(3-(trifluoromethoxy)phenyl)pyrazolo[1,5-a][1,3,5]triazine-8-carbaldehyde (71 mg, 75%) as a tan solid. LCMS (ES): >95% pure, m/z 364 [M+1]⁺.

(Z)-5-((4-(cyclopropylamino)-2-(3-(trifluoromethoxy)phenyl)pyzazolo[1,5-a][1,3,5]triazine-8-yl)methylene)imidazolidine-2,4-dione was prepared using chemistries similar to those exemplified in Example 197. LCMS (ES): >90% pure, m/z 446 [M+1]⁺.

The following compounds were prepared using chemistries described in Example 199, Example 200, Example 201, Example 202, Example 203, Example 204, Example 205, Example 206, Example 207 and Example 208; using appropriate reagents. General methods for preparation of such compounds are included in Figures 3-14 herein. Reagents bearing two reactive amino groups were generally used as mono-Boc protected. The protecting group was removed by reaction with trifluoroacetic acid in methylene chloride prior to purification. Compounds were isolated by filtration after addition of water or methanol. Some compounds were purified by preparative HPLC and isolated as TFA salts after evaporation at the Genevac. Compounds were characterized by LCMS. Table 33B shows the biological activities of the compounds listed in Table 33.A. Compound CK2: IC50 (µM) PIM2: IC50 (5 µM ATP) AB: MDAMD453 (µM) AB: BxPC3 (µM) H14 <1.0 1.3064 25.377 22.762 I14 <0.01 > 2.5000 >30 >30 J14 <0.01 > 2.5000 1.193 5.747 K14 <0.1 > 2.5000 L14 <0.01 > 2.5000 1.003 11.197 M14 <0.01 > 2.5000 0.77 2.905 N14 <0.01 2.3344 1.998 1.766 O14 <0.01 > 2.5000 0.753 2.178 P14 <0.01 > 2.5000 3.129 >30 Q14 <0.01 > 2.5000 1.06 2.671 R14 <0.01 2.4058 1.602 15.41 S14 <0.1 > 2.5000 11.279 11.668 T14 <0.1 1.25 U14 <0.1 > 2.5000 V14 <1.0 > 2.5000 W14 <0.1 > 2.5000 6.386 23.384 X14 <0.01 1.1205 0.559 1.517 Y14 <0.01 > 2.5000 3.325 6.748 A15 >1 > 2.5000 B15 <0.01 > 2.5000 2.214 3.394 C15 <1.0 > 2.5000 D15 <0.1 1.5464 E15 <1.0 > 2.5000 F15 <1.0 > 2.5000 G15 <0.1 > 2.5000 H15 <0.1 > 2.5000 I15 <0.1 > 2.5000 J15 <0.01 > 2.5000 1.936 11.99 K15 <0.1 1.6558 5.319 23.688 L15 <0.1 > 2.5000 M15 >1 > 2.5000 N15 >1 > 2.5000 O15 >1 > 2.5000 P15 <1.0 > 2.5000 Q15 <1.0 > 2.5000 R15 <1.0 > 2.5000 S15 >1 > 2.5000 T15 <1.0 > 2.5000 U15 <1.0 > 2.5000 V15 <1.0 > 2.5000 W15 <1.0 > 2.5000 X15 <1.0 18.336 14.152 Y15 <0.1 > 2.5000 6.603 10.731 Z15 <0.01 > 2.5000 6.283 15.114 A16 <0.1 > 2.5000 17.82 > 30 B16 <0.01 > 2.5000 2.627 5.528 C16 <0.1 1.7695 1.859 1.841 D16 <0.1 > 2.5000 5.03 17.83 E16 <0.01 > 2.5000 7.856 11.797 F16 <0.01 > 2.5000 3.545 17.187 G16 <0.01 > 2.5000 5.177 19.446 H16 <0.1 > 2.5000 18.952 >30 I16 <0.1 > 2.5000 12.508 12.673 J16 <0.1 > 2.5000 11.685 18.88 K16 <1.0 > 2.5000 9.2 12.494 L16 <0.1 > 2.5000 9.964 11.216 M16 <0.01 5.059 9.052 N16 <1.0 25.644 > 30 O16 <0.1 7.829 >30 P16 <0.1 10.353 > 30 Q16 <0.1 17.611 > 30 R16 <0.1 13.114 19.2 S16 <0.01 > 2.5000 4.697 8.049 T16 <0.01 1.9391 2.491 1.712 U16 <0.0.1 > 2.5000 4.313 8.232 V16 <0.01 2.438 5.738 5.492 W16 <0.1 > 2.5000 27.524 9.27 X16 <1.0 > 2.5000 17.619 9.053 Y16 <0.1 > 2.5000 Z16 <0.01 <1.0 14.666 2.909 A17 <0.1 1.8629 12.569 5.872 B17 <0.01 > 2.5000 12.517 6.862 C17 <0.1 > 2.5000 11.841 2.389 D17 <0.01 1.884 3.353 8.144 E17 <1.0 > 2.5000 F17 <0.01 > 2.5000 3.97 15.62 G17 <0.1 <1.0 H17 <0.1 <1.0 6.919 6.611 I17 <0.01 1.1903 0.442 7.667 J17 <0.01 > 2.5000 1.416 9.445 K17 <0.1 > 2.5000 L17 <0.1. 2.0781 20.449 5.948 M17 <0.01 > 2.5000 27.014 > 30 N17 <0.1 > 2.5000 13.419 19.135 O17 <0.1 > 2.5000 12.459 17.049 P17 <0.1 > 2.5000 Q17 <0.1 1.6281 > 30 2.009 R17 <1.0 > 2.5000 S17 <0.1 > 2.5000 2.594 6.849 T17 <0.01 <1.0 2.751 4.95 U17 <0.1 > 2.5000 4.554 10.817 V17 <0.1 > 2.5000 13.033 1.55 W17 <0.1 > 2.5000 X17 <0.1 > 2.5000 2.477 5.229 Y17 <0.1 > 2.5000 8.283 3.409 Z17 <0.1 > 2.5000 4.486 9.071 A18 <0.01 > 2.5000 1.478 4.729 B18 <0.01 1.304 2.64 4.027 C18 <0.01 > 2.5000 1.245 1.388 D18 <0.01 > 2.5000 2.286 4.684 E18 <0.1 > 2.5000 10.575 9.784 F18 <0.01 > 2.5000 >30 >30 G18 <0.1 > 2.5000 13.472 14.435 H18 <0.01 > 2.5000 1.907 4.537 I18 <0.1 > 2.5000 8.221 14.522 J18 <0.1 > 2.5000 K18 <0.1 > 2.5000 L18 <0.1 > 2.5000 M18 <0.1 > 2.5000 N18 <0.1 > 2.5000 26.829 25.022 O18 <0.01 > 2.5000 13.2 > 30 P18 <0.1 > 2.5000 21.799 > 30 Q18 <0.1 > 2.5000 1.368 8.975 R18 <0.1 > 2.5000 8.555 13.435 S18 <0.1 > 2.5000 T18 <0.01 > 2.5000 3.136 4.168 U18 <0.1 > 2.5000 V18 <0.1 > 2.5000 W18 <0.01 > 2.5000 11.668 > 30 X18 <0.01 > 2.5000 13.548 18.585 Y18 <0.1 > 2.5000 18.328 5.013 Z18 <0.01 > 2.5000 11.97 >30 A19 <0.1 > 2.5000 B19 <0.1 > 2.5000 C19 <0.01 > 2.5000 D19 <0.1 > 2.5000 E19 <0.1 > 2.5000 F19 <0.01 > 2.5000 G19 <0.01 > 2.5000 H19 <0.1 > 2.5000 I19 <0.1 > 2.5000 J19 <0.01 > 2.5000 K19 <0.1 > 2.5000 L19 <0.1 > 2.5000 M19 <0.01 N19 <0.1 O19 <0.1 P19 <0.01 Q19 <0.01 R19 <0.01 S19 <0.1

The compounds described in the following table can be prepared using chemistries described on Figure 7.

The following molecules can be prepared using chemistries similar to Example 206, Example 207 and Example 208.

The molecules described in Figure 8 were prepared using chemistries described in Example 201, using bases such as K₂CO₃ or sodium hydride.

The chemistry described on Figure 9 can be used to prepare analogs of formula 11. 4-bromo-6-chloropyridazin-3-amine 1 can be reacted with 2 using conditions analogous to the preparation described in the patent application WO2009/100375 to form compound 3. Compound 3 can react with amine R₈R₇NH to form compound 4. Compound 4 can be transformed to compound 5 by nucleophilic substitutions with amines, anilines, alcohols, phenols or thiophenols, in the presence of a base, or by transition metal catalyzed conversions such as Suzuki coupling with boronic acid or esters of formula WB(OR.)₂. Compound 5 can be transformed to compound 6 by reduction with LiAlH₄. Alcohol 6 can be converted to aldehyde 7 by oxidation with DCC or under Swern conditions. Compound 5 can react with an organometallic reagent exemplified by Grignard reagent R⁴MgX to form secondary alcohol 8. This compound can be converted to alkylketone 9 under conditions analogous to the conditions used to convert 6 into 7. Compounds 7 and 9 can both be converted to compound 11 by condensation with 10 in a solvent such as ethanol and in the presence of a base such as piperidine.

The compounds described in the following table can be prepared using chemistry described on Figure 9.

Figures 10-14 illustrate other synthesis methods that can be used to prepare compounds of the invention.

Step A. To 5,7-dichloropyrazolo[1,5-a]pyrimidine (200 mg, 1.06 mmol) in acetonitrile was added Et₃N (148 µl, 1.06 mmol) and cyclopropylamine (75 µl, 1.06 mmol). The reaction was heated at 80°C overnight. The mixture was concentrated under reduced pressure, dissolved in dichloromethane, and washed with water. The resulting organic layer was dried over Na₂SO₄ and concentrated under reduced-pressure to afford 156 mg of 5-chloro-N-cyclopropylpyrazolo[1,5-a]pyrimidin-7-amine (70% yield). LCMS (M+1=209)

Step B. To 5-chloro-N-cyclopropylpyrazolo[1,5-a]pyrimidin-7-amine (156 mg, 0.75 mmol) in DMF was added POCl₃ (205 µl, 2.25 mmol). The mixture was stirred at room temperature for 3 hours. Ice was added to quench POCl₃, and then the mixture was neutralized with 1 M NaOH. Dichloromethane was added and the product was extracted three times. The organic layer was dried over Na₂SO₄ and concentrated under reduced pressure to yield 5-chloro-7-(cyclopropylamino)pyrazolo[1,5-a]pyrimidine-3-carbaldehyde. Some residual DMF could not be removed. LCMS (M+1=237)

Step C. To 5-chloro-7-(cyclopropylamino)pyrazolo[1,5-a]pyrimidine-3-carbaldehyde (177 mg, 0.75 mmol) in 1,4-dioxane was added 3-chloroaniline (397 µl, 3.75 mmol). The mixture was heated in microwave at 1.20°C for 60 minutes. Precipitate was filtered off, and the filtrate was prepared by TLC (1% methanol/dichloromethane) to yield 26 mg (11% yield) of 5-(3-chloxophenylamino)-7-(cyclopropylamino)pyrazolo[1,5-a]pyrimidine-3-carbaldehyde. LCMS (M+1=328)

Step D. To 5-(3-chlorophenylamino)-7-(cyclopropylamino)pyrazolo[1,5-a]pyrimidine-3-carbaldehyde (26 mg, 0.08 mmol) in EtOH was added hydantoin (8 mg, 0.08 mmol) and piperdine (8 µl, 0.08 mmol). The mixture was stirred at 70°C for 3 days. Insolubles were filtered off, and filtrate was concentrated under reduced pressure. Filtrate was then dissolved in methanol and purified by HPLC to yield 5-(5-(3-chlorophenylamino)-7-(cyclopropylamino)pyrazolo[1,5-a]pyrimidin-3-yl)methylene)imidazolidine-2,4-dione. LCMS (M+1=410)

Step A. To 5-chloro-7-(cyclopropylamino)pyrazolo[1,5-a]pyrimidine-3-carbaldehyde (440 mg, 1.86 mmol) in EtOH was added thiazolidine-2,4-dione (458 mg, 3.91 mmol) and piperidine (208 µl, 2.05 mmol). The reaction was heated at 80°C overnight. 3 mL Isopropanol was added in the morning, along with 218 mg thiazolidine-2,4-dione, 94 µL piperdine. Temperature was increased to 90°C and left overnight. Precipitate was filtered while hot and dissolved in methanol. 1 mL of 1M HCl was added and the mixture sonicated. Precipitate was filtered and washed with methanol to yield 340 mg (54% yield) 5-((5-chloro-7-(cyclopropylamino)pyrazolo[1,5-a]pyrimidin-3-yl)methylene)thiazolidine-2,4-dione as a yellow powder. LCMS (M+1=336)

Step B. To 5-((5-chloro-7-(cyclopropylamino)pyrazolo[1,5-a]pyrimidin-3-yl)methylene)thiazolidine-2,4-dione (30 mg, 0.09 mmol) in N-methylpyrrolidinone (NMP) was added 2-methylpropan-1-amine (20 mg, 0.268 mmol). The reaction was heated at 130°C overnight. Mixture was diluted with methanol and prepared by HPLC to yield 5-((7-(cyclopropylamino)-5-(isobutylamino)pyrazolo[1,5-a]pyrimidin-3-yl)methylene)thiazolidine-2,4-dione. LCMS (M+1=373)

The titled compound was prepared using a method analogous to that described for Example 210. LCMS (M+1=375)

The titled compound was prepared using a method analogous to that described for Example 210. LCMS (M+1=373)

The titled compound was prepared using a method analogous to that described for Example 210. LCMS (M+1=345)

The titled compound was prepared using a method analogous to that described for Example 210. LCMS (M+1=414)

The titled compound was prepared using a method analogous to that described for Example 210. LCMS (M+1=363)

The titled compound was prepared using a method analogous to that described for Example 210. LCMS (M+1=414)

The titled compound was prepared using a method analogous to that described for Example 210. LCMS (M+1=416)

To 5-((5-chloro-7-(cyclopropylamino)pyrazolo[1,5-a]pyrimidin-3-yl)methylene)thiazolidine-2,4-dione (20 mg, 0.06 mmol) in NMP was added 3-chloroaniline (38 µL, 0.36 mmol) and few granules p-toluenesulfonic acid. The reaction was heated in microwave at 180°C for 1.5 hours. Mixture was filtered and prepared by HPLC then preparative TLC (1% methanol/dichloromethane) to yield 5-((5-(3-chlorophenylamino)-7-(cyclopropylamino)pyrazolo[1,5-a]pyrimidin-3-yl)methylene)thiazolidine-2,4-dione as a yellow solid. LCMS (M+1=427)

Step A. 7-(Cyclopropylamino)-2,5-dimethylpyrazolo[1,5-a]pyrimidine-3-carbaldehyde was prepared from N-cyclopropyl-2,5-dimethylpyrazolo[1,5-a]pyrimidin-7-amine using methods analogous to those described in Example 209, Step B. LCMS (M+1=231)

Step B. To 7-(cyclopropylamino)-2,5-dimethylpyrazolo[1,5-a]pyrimidine-3-carbaldehyde (0.25 mmol) in DMF was added thiazolidine-2,4-dione (88 mg, 0.75 mmol) and piperidine (25 µl, 0.25 mmol). The mixture was stirred at room temperature overnight. Mixture was prepared by HPLC to yield 5-((7-(cyclopropylamino)-2,5-dimethy)pyrazolo[1,5-a]pyrimidin-3-yl)methylene)thiazolidine-2,4-dione. LCMS (M+1=330)

Step A. 7-(Cyclopropylamino)pyrazolo[1,5-a]pyrimidine-3-carbaldehyde was prepared from N-cyclopropylpyrazolo[1,5-a]pyrimidin-7-amine using the methods described in Example 219, Step A. LCMS (M+1=203)

Step B. The titled compound was prepared from 7-(cyclopropylamino)pyrazolo[1,5-a]pyrimidine-3-carbaldehyde using methods analogous to those described in Example 219, Step B, except the product was isolated by filtration, washed with methanol, and air dried. LCMS (M+1=302)

Step A. To 5-chloro-7-(cyclopropylamino)pyrazolo[1,5-a]pyrimidine-3-carbaldehyde (400 mg, 1.70 mmol) in EtOH was added hydantoin (86mg, 1.86mmol) and pyrrolidine (14µL, 0.17mmol). The reaction was stirred at 70°C over weekend. Precipitate was filtered and air dried to yield 180mg (33% yield) 5-((5-chloro-7-(cyclopropylamino)pyrazolo-[1,5-a]pyrimidin-3-yl)methylene)imidazolidine-2,4-dione. LCMS (M+1=319)

Step B. To 5-((5-chloro-7-(cyclopropylamino)pyrazolo[1,5-a]pyrimidin-3-yl)methylene)imidazolidine-2,4-dione (30 mg, 0.09 mmol) in 1,4-dioxane was added 1-(pyridin-2-yl)piperazine (58 µL, 4.10 mmol) and Et₃N (13µL, 0.09 mmol). Reaction was then heated 120°C for 35 minutes in microwave. Solvent was removed under reduced pressure, and mixture was dissolved in methanol. Solid was isolated by filtration, then air dried to yield 11 mg (26% yield) 5-((7-(cyclopropylamino)-5-(4-(pyridin-2-yl)piperazin-1-yl)pyrazolo[1,5-a]pyrimidin-3-yl)methylene)imidazolidine-2,4-dione. LCMS (M+1=446)