THIOZOLIDINEDIONE DERIVATIVES AS PI3 KINASE INHIBITORS

Field of the invention

This invention relates to the use of thiozolidinedione derivatives for the modulation, notably the inhibition of the activity or function of the phosphoinositide 3' OH kinase family (hereinafter PB kinases), suitably, PBKα, PBKδ, PBKβ, and/or PBKγ, particularly PBKα. Suitably, the present invention relates to the use of thiozolidinedione derivatives in the treatment of one or more disease states selected from: autoimmune disorders, inflammatory diseases, cardiovascular diseases, neurodegenerative diseases, allergy, asthma, pancreatitis, multiorgan failure, kidney diseases, platelet aggregation, cancer, sperm motility, transplantation rejection, graft rejection and lung injuries, particularly cancer.

Background of the invention

Cellular membranes represent a large store of second messengers that can be enlisted in a variety of signal transduction pathways. In regards function and regulation of effector enzymes in phospholipids signaling pathways, these enzymes generate second messengers from the membrane phospholipid pools (class I PB kinases (e.g. PBKalpha) are dual-specificity kinase enzymes, meaning they display both: lipid kinase (phosphorylation of phosphoinositides) as well as protein kinase activity, shown to be capable of phosphorylation of protein as substrate, including auto-phosphorylation as intramolecular regulatory mechanism. These enzymes of phospholipids signaling are activated in response to a variety of extra-cellular signals such as growth factors, mitogens, integrins (cell-cell interactions) hormones, cytokines, viruses and neurotransmitters such as described in Scheme I hereinafter and also by intracellular regulation by other signaling molecules (cross-talk, where the original signal can activate some parallel pathways that in a second step transmit signals to PBKs by intra-cellular signaling events), such as small GTPases, kinases or phosphatases for example. Intracellular regulation can also occur as a result of aberrant expression or lack of expression of cellular oncogenes or tumor suppressors. The inositol phospholipid (phosphoinositides) intracellular signaling pathways begin with activation of signaling molecules (extra cellular ligands, stimuli, receptor dimerization, transactivation by heterologous receptor (e.g.

receptor tyrosine kinase) and the recruitment and activation of PBK including the involvement of G-protein linked transmembrane receptor integrated into the plasma membrane.

PBK converts the membrane phospholipid PI(4,5)P ₂ into PI(3,4,5)P ₃ that functions as a second messenger. PI and PI(4)P are also substrates of PBK and can be phosphorylated and converted into PBP and PI(3,4)P ₂ , respectively. In addition, these phosphoinositides can be converted into other phosphoinositides by 5'- specific and 3 '-specific phophatases, thus PBK enzymatic activity results either directly or indirectly in the generation of two 3' -phosphoinositide subtypes that function as 2 ^nd messengers in intra-cellular signal transduction pathways (Trends Biochem. Sci. 22(7) p.267-72 (1997) by Vanhaesebroeck et al: Chem. Rev. 101(8) p.2365-80 (2001) by Leslie et al (2001); Annu. Rev. Cell.Dev. Biol. 17p, 615-75 (2001) by Katso et al. and Cell. MoI. Life Sci. 59(5) p.761-79 (2002) by Toker et al.). Multiple PBK isoforms categorized by their catalytic subunits, their regulation by corresponding regulatory subunits, expression patterns and signaling-specific functions (pi 10a, β, δ and γ) perform this enzymatic reaction (Exp. Cell. Res. 25 (1) p. 239-54 (1999) by Vanhaesebroeck and Katso et al., 2001, above).

The closely related isoforms pi 10a and β are ubiquitously expressed, while δ and γ are more specifically expressed in the haematopoietic cell system, smooth muscle cells, myocytes and endothelial cells (Trends Biochem. Sci. 22(7) p.267-72 (1997) by Vanhaesebroeck et al.). Their expression might also be regulated in an inducible manner depending on the cellular, tissue type and stimuli as well as disease context. Inducibility of protein expression includes synthesis of protein as well as protein stabilization that is in part regulated by association with regulatory subunits.

To date, eight mammalian PBKs have been identified, divided into three main classes (I, II, and III) on the basis of sequence homology, structure, binding partners, mode of activation, and substrate preference. In vitro, class I PBKs can phosphorylate phosphatidylinositol (PI), phosphatidylinositol-4-phosphate (PI4P), and phosphatidylinositol-4,5-bisphosphate (PI(4,5)P ₂ ) to produce phosphatidylinositol-3-phosphate (PBP), phosphatidylinositol-3,4-bisphosphate (PI(3,4)P ₂ , and phosphatidylinositol-3,4,5-trisphosphate (PI(3,4,5)P ₃ , respectively. Class II PBKs phosphorylate PI and phosphatidylinositol-4-phosphate. Class III PBKs can only phosphorylate PI (Vanhaesebrokeck et al., 1997, above;

Vanhaesebroeck et al., 1999, above and Leslie et al, 2001, above)

Scheme I: Conversion of PI(4,5)P2 to PIP3

PtdIns(4,5)P ₂

PtdIns(3,4,5)P ₃

As illustrated in Scheme A above, phosphoinositide 3-kinases (PBKs) phosphorylate the hydroxyl of the third carbon of the inositol ring. The phosphorylation of phosphoinositides that generate Ptdlns to 3,4,5-trisphosphate (PtdIns(3,4,5)P3), PtdIns(3,4)P2 and PtdIns(3)P produce second messengers for a variety of signal transduction pathways, including those essential to cell proliferation, cell differentiation, cell growth, cell size, cell survival, apoptosis, adhesion, cell motility, cell migration, chemotaxis, invasion, cytoskeletal rearrangement, cell shape changes, vesicle trafficking and metabolic pathway (Katso et al., 2001, above and MoL Med. Today 6(9) p. 347-57 (2000) by Stein). G-protein coupled receptors mediate phosphoinositide 3'OH-kinase activation via small

GTPases such as Gβγ and Ras, and consequently PBK signaling plays a central role in establishing and coordinating cell polarity and dynamic organization of the cytoskeleton - which together provides the driving force of cells to move. Chemotaxis - the directed movement of cells toward a concentration gradient of chemical attractants, also called chemokines is involved in many important diseases such as inflammation/auto-immunity, neuro degeneration, antiogenesis, invasion/metastasis and wound healing (Immunol. Today 21(6) p. 260-4 (2000) by Wyman et al.; Science 287(5455) p. 1049-53 (2000) by Hirsch et al; FASEB J. 15(11) p. 2019-21 (2001) by Hirsch et al. and Nat. Immunol. 2(2) p. 108-15 (2001) by Gerard et al.).

Advances using genetic approaches and pharmacological tools have provided insights into signalling and molecular pathways that mediate chemotaxis in response to chemoattractant activated G-protein coupled receptors. PB -Kinase, responsible for generating these phosphorylated signalling products, was originally identified as an activity associated with viral oncoproteins and growth factor receptor tyrosine kinases that phosphorylates phosphatidylinositol (PI) and its phosphorylated derivatives at the 3'-hydroxyl of the inositol ring (Panayotou et al., Trends Cell Biol. 2 p. 358-60 (1992)). However, more recent biochemical studies revealed that class I PB kinases (e.g. class IB isoform PBKγ) are dual-specific kinase enzymes, meaning they display both lipid kinase and protein kinase activity, shown to be capable of phosphorylation of other proteins as substrates, as well as auto-phosphorylation as an intra-molecular regulatory mechanism.

PB -kinase activation, is therefore believed to be involved in a range of cellular responses including cell growth, differentiation, and apoptosis (Parker et al., Current Biology, 5 p. 577-99 (1995); Yao et al., Science, 267 p. 2003-05 (1995)). PB -kinase appears to be involved in a number of aspects of leukocyte activation. A p85-associated PB-kinase activity has been shown to physically associate with the cytoplasmic domain of CD28, which is an important costimulatory molecule for the activation of T-cells in response to antigen (Pages et al., Nature, 369 p. 327-29 (1994); Rudd, Immunity 4 p. 527-34 (1996)). Activation of T cells through CD28 lowers the threshold for activation by antigen and increases the magnitude and duration of the proliferative response. These effects are linked to increases in the transcription of a number of genes including interleukin-2 (IL2), an important T cell growth factor (Fraser et al., Science 251 p. 313-16 (1991)). Mutation of CD28 such that it can no longer interact with PB-kinase leads to a failure to initiate IL2

production, suggesting a critical role for PI3-kinase in T cell activation. PBKγ has been identified as a mediator of G beta-gamma-dependent regulation of JNK activity, and G beta-gamma are subunits of heterotrimeric G proteins (Lopez -Ilasaca et al, J. Biol. Chem. 273(5) p. 2505-8 (1998)). Cellular processes in which PBKs play an essential role include suppression of apoptosis, reorganization of the actin skeleton, cardiac myocyte growth, glycogen synthase stimulation by insulin, TNFα- mediated neutrophil priming and superoxide generation, and leukocyte migration and adhesion to endothelial cells.

Recently, (Laffargue et al., Immunity 16(3) p. 441-51 (2002)) it has been described that PBKγ relays inflammatory signals through various G(i)-coupled receptors and its central to mast cell function, stimuli in context of leukocytes, immunology includes cytokines, chemokines, adenosines, antibodies, integrins, aggregation factors, growth factors, viruses or hormones for example (J. Cell. Sci. 114(Pt 16) p. 2903-10 (2001) by Lawlor et al.; Laffargue et al., 2002, above and Curr. Opinion Cell Biol. 14(2) p. 203-13 (2002) by Stephens et al.).

Specific inhibitors against individual members of a family of enzymes provide invaluable tools for deciphering functions of each enzyme. Two compounds, LY294002 and wortmannin (cf. hereinafter), have been widely used as PB-kinase inhibitors. These compounds are non-specific PBK inhibitors, as they do not distinguish among the four members of Class I PB -kinases. For example, the IC50 values of wortmannin against each of the various Class I PB -kinases are in the range of 1-10 nM. Similarly, the IC50 values for LY294002 against each of these PB-kinases is about 15-20 μM (Fruman et al., Ann. Rev. Biochem., 67, p. 481-507 (1998)), also 5-10 microM on CK2 protein kinase and some inhibitory activity on phospholipases. Wortmannin is a fungal metabolite which irreversibly inhibits PBK activity by binding covalently to the catalytic domain of this enzyme. Inhibition of PBK activity by wortmannin eliminates subsequent cellular response to the extracellular factor. For example, neutrophils respond to the chemokine fMet-Leu- Phe (fMLP) by stimulating PBK and synthesizing Ptdlns (3, 4, 5)P3. This synthesis correlates with activation of the respirators burst involved in neutrophil destruction of invading microorganisms. Treatment of neutrophils with wortmannin prevents the fMLP-induced respiratory burst response (Thelen et al., Proc. Natl. Acad. Sci. USA, 91, p. 4960-64 (1994)). Indeed, these experiments with wortmannin, as well as other experimental evidence, shows that PBK activity in cells of hematopoietic lineage, particularly neutrophils, monocytes, and other types of leukocytes, is

involved in many of the non-memory immune response associated with acute and chronic inflammation.

LY294002 Wortmannin

Based on studies using wortmannin, there is evidence that PB -kinase function is also required for some aspects of leukocyte signaling through G-protein coupled receptors (Thelen et al., 1994, above). Moreover, it has been shown that wortmannin and LY294002 block neutrophil migration and superoxide release. Cyclooxygenase inhibiting benzofuran derivatives are disclosed by John M. Janusz et al., in J. Med. Chem. 1998; Vol. 41, No. 18.

It is now well understood that deregulation of onocogenes and tumour- suppressor genes contributes to the formation of malignant tumours, for example by way of increase cell growth and proliferation or increased cell survival. It is also now known that signaling pathways mediated by the PBK family have a central role in a number of cell processes including proliferation and survival, and deregulation of these pathways is a causative factor a wide spectrum of human cancers and other diseases (Katso et al., Annual Rev. Cell Dev. Biol. 2001, j/7: 615-617 and Foster et al, J. Cell Science. 2003, JJ6: 3037-3040). Class I PBK is a heterodimer consisting of a pi 10 catalytic subunit and a regulatory subunit, and the family is further divided into class Ia and Class Ib enzymes on the basis of regulatory partners and mechanism of regulation. Class Ia enzymes consist of three distinct catalytic subunits (pi 10a, pi lOβ, and pi lOδ) that dimerise with five distinct regulatory subunits (p85α, p55α, p50α, p85β, and p55γ), with all catalytic subunits being able to interact with all regulatory subunits to form a variety of heterodimers. Class Ia PBK are generally activated in response to growth factor-stimulation of receptor tyrosine kinases, via interaction of the regulatory subunit SH2 domains with specific phospho-tyrosine residues of the

activated receptor or adaptor proteins such as IRS-I. Small GTPases (ras as an example) are also involved in the activation of PBK in conjunction with receptor tyrosine kinase activation. Both pi 10a and pi lOβ are constitutively expressed in all cell types, whereas pi lOδ expression is more restricted to leukocyte populations and some epithelial cells. In contrast, the single Class Ib enzyme consists of a pi lOγ catalytic subunit that interacts with a plOl regulatory subunit. Furthermore, the Class Ib enzyme is activated in response to G-protein coupled receptor (GPCR) systems and its expression appears to be limited to leukocytes.

There is now considerable evidence indicating that Class Ia PBK enzymes contribute to tumourigenesis in a wide variety of human cancers, either directly or indirectly (Vivanco and Sawyers, Nature Reviews Cancer, 2002, 2, 489-501). For example, the pi 10a subunit is amplified in some tumours such as those of the ovary (Shayesteh, et al, Nature Genetics. 1999, 21 : 99-102) and cervix (Ma et al, Oncogene, 2000, 19: 2739-2744). More recently, activating mutations within pi 10a (PIK3CA gene) have been associated with various other tumors such as those of the colon and of the breast and lung (Samuels, et al., Science, 2004, 304, 554). Tumor- related mutations in p85α have also been identified in cancers such as those of the ovary and colon (Philp et al, Cancer Research, 2001, 61_, 7426-7429). In addition to direct effects, it is believed that activation of Class Ia PBK contributes to tumourigenic events that occur upstream in signaling pathways, for example by way of ligand-dependent or ligand-independent activation of receptor tyrosine kinases, GPCR systems or integrins (Vara et al, Cancer Treatment Reviews, 2004, 30, 193- 204). Examples of such upstream signaling pathways include over-expression of the receptor tyrosine kinase Erb2 in a variety of tumors leading to activation of PBK- mediated pathways (Harari et al., Oncogene, 2000, \9_, 6102-6114) and over- expression of the oncogene Ras (Kauffmann-Zeh et al., Nature, 1997, 385, 544- 548). In addition, Class Ia PBKs may contribute indirectly to tumourigenesis caused by various downstream signaling events. For example, loss of function of the PTEN tumor-suppressor phosphatase that catalyses conversion of PI(3,4,5)P3 back to PI(4,5)P2 is associated with a very broad range of tumors via deregulation of PBK- mediated production of PI(3,4,5)P3 (Simpson and Parsons, Exp. Cell Res., 2001, 264, 29-41). Furthermore, augmentation of the effects of other PBK-mediated signaling events is believed to contribute to a variety of cancers, for example by activation of AKT (Nicholson and Andeson, Cellular Signaling, 2002, U, 381-395).

In addition to a role in mediating proliferative and survival signaling in tumor cells, there is also good evidence that class Ia PBK enzymes also contributes

to tumourigenesis via its function in tumor-associated stromal cells. For examples, PBK signaling is known to play an important role in mediating angiogenic events in endothelial cells in response to pro-angiogenic factors such as VEGF (abid et al., Arterioscler. Thromb. Vase. Biol.. 2004, 24, 294-300). As Class I PBK enzymes are also involved in motility and migration (Sawyer, Expert Opinion investing. Drugs, 2004, j_3, 1-19), PBK inhibitors are anticipated to provide therapeutic benefit via inhibition of tumor cell invasion and metastasis.

Summary of the Invention

This invention relates to novel compounds of Formula (I):

(I)

wherein

in which Rl is selected from: hydrogen, Cl-C6alkyl, substituted Cl-C6alkyl, aryl, substituted aryl, arylalkyl, substituted arylalkyl;

R2 and R3 are independently selected from: hydrogen, halogen, Cl-C6acyl, amino, substituted Cl-C6alkyl, Cl-C6alkyl, C3-C7cycloalkyl, substituted C3- C7cycloalkyl, C3-C7heterocycloalkyl, substituted C3-C7heterocycloalkyl, aminoalkyl, substituted aminoalkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, arylalkyl, substituted arylalkyl, arylcycloalkyl, substituted arylcycloalkyl, heteroarylalkyl, substituted heteroarylalkyl, arylaminoalkyl, arylaminoheterocycloalkyl(CH2) _n -, cyano, hydroxyl, alkoxy, aryloxy, N- acylamino, acyloxy, arylamino, nitro, CO2R15, and CONR20R25, where R15, R20 and R25 are independently selected from: hydrogen and alkyl; n is 0-3, m is 0-2;

A, B, D, E, and G together form a ring containing from 1 to 2 double bonds and from 1 to 4 nitrogens; and

X, Y, Z are each independently selected from CH, CR3, or N wherein R3 is as defined above; provided that one and only one of A and B is N; and/or a pharmaceutically acceptable salt, hydrate, solvate or pro-drug thereof.

This invention also relates to a method of treating cancer, which comprises administering to a subject in need thereof an effective amount of a compound of Formula (I).

This invention also relates to a method of treating one or more disease states selected from: autoimmune disorders, inflammatory diseases, cardiovascular diseases, neurodegenerative diseases, allergy, asthma, pancreatitis, multiorgan failure, kidney diseases, platelet aggregation, sperm motility, transplantation rejection, graft rejection and lung injuries, which comprises administering to a subject in need thereof an effective amount of a compound of Formula (I).

Also invented is a novel process of making thiazolidineione derivatives. Included in the present invention are methods of co-administering the present PB kinase inhibiting compounds with further active ingredients.

Detailed Description of the Invention

Present compounds of Formula (I) inhibit PB kinases. Suitably, the compounds of Formula (I) inhibit one or more PB kinases selected from: PBKα, PBKδ, PBKβ and PBKγ.

This invention also relates to a compound of Formula (I), wherein Rl is hydrogen, or Cl-C6alkyl.

This invention also relates to a compound of Formula (I), wherein Rl is hydrogen.

This invention also relates to a compound of Formula (I), wherein X and Y are, CH or CR3; G and E are, CH or CR2; and/or a pharmaceutically acceptable salt, hydrate, solvate or pro-drug thereof.

This invention also relates to a compound of Formula (I), wherein X, Y, and E are CH; G is CR2, A is C, and B is N.

This invention also relates to a compound of Formula (I), wherein X, Y, and E are CH; G is CR2, A is C, and B and D are N.

This invention also relates to a compound of Formula (I), wherein X, Y, and E are CH; G is CR2, A is C, and B, D and Z are N.

This invention also relates to a compound of Formula (I), wherein X, Y, and E are CH; G is CR2, A is C, B and D are N; R2 is selected from a group consisting of: Cl-C6acyl, aminoalkyl, substituted aminoalkyl, arylaminoalkyl, arylaminoheterocycloalkyl(CH2) _n -, heteroaryl, substituted heteroaryl, CO ₂ R ₁₅ , and CONR20R25; and Ri ₅ , where R2, R ₂ o and R25 are defined according to claiml, and n is 0-3.

This invention also relates to a compound of Formula (I), wherein Y and E are CH; G is CR2, and A is N.

This invention also relates to a compound of Formula (I), wherein Y and E are CH; G is CR2, and A, D, and Z are N.

This invention also relates to a compound of Formula (I), wherein Y and E are CH; G is CR2, and A and D are N. This invention also relates to a compound of Formula (I), wherein Y and E are CH; G is CR2, and A, D and X are N.

This invention also relates to a compound of Formula (I), wherein Y and E are CH; G is CR2, R2 is Cl-C6acylalkyl, substituted or unsubstituted heteroaryl, CO ₂ Ri ₅ , and CONR ₂₀ R ₂₅ and A is N. Unless otherwise defined, R2, R3 as used herein are as decribed for compounds of Formula (I) above.

Included among the compounds of Formula (I) are compounds useful as inhibitors of PI3 kinase activity selected from:

Ethyl 6-[(Z)-(2,4-dioxo-l,3-thiazolidin-5-ylidene)methyl]imidazo[l ,2- α]pyridine-3-carboxylate,

6-[(Z)-(2,4-dioxo-l,3-thiazolidin-5-ylidene)methyl]-λ/-e thylimidazo[l,2- α]pyridine-3-carboxamide,

(5Z)-5- {[3-(4-pyridinyl)imidazo[ 1 ,2-α]pyridin-6-yl]methylidene} -1,3- thiazolidine-2,4-dione sodium salt,

(5Z)-5-(imidazo[ 1 ,2-α]pyridin-6-ylmethylidene)- 1 ,3-thiazolidine-2,4- dione,

(5Z)-5-[(3-acetylimidazo[l ,2-α]pyridin-6-yl)methylidene]-l ,3- thiazolidine-2,4-dione,

(5Z)-5-(imidazo[ 1 ,2-α]pyrimidin-6-ylmethylidene)- 1 ,3-thiazolidine-2,4- dione,

(5Z)-5-[(3-acetyl-2-methylimidazo[l,2-α]pyridin-6-yl)met hylidene]-l,3- thiazolidine-2,4-dione,

(5Z)-5 - { [3 -(trifluoromethyl)imidazo [ 1 ,2-α]pyridin-6-yl]methylidene } - 1 ,3 - thiazolidine-2,4-dione,

6-[(Z)-(2,4-dioxo-l,3-thiazolidin-5-ylidene)methyl]imidaz o[l,2- α]pyridine-3-carbonitrile,

(5Z)-5-{[3-(2,2-dimethylpropanoyl)imidazo[l,2-α]pyridin- 6- yl]methylidene} - 1 ,3-thiazolidine-2,4-dione,

(5Z)-5-[(3-propanoylimidazo[l,2-α]pyridin-6-yl)methylide ne]-l,3- thiazolidine-2,4-dione,

2-{6-[(Z)-(2,4-dioxo-l,3-thiazolidin-5-ylidene)methyl]imi dazo[l,2- α]pyridin-3-yl} -2-oxoethyl acetate,

(5Z)-5 - { [3 -( 1 -hydroxypropyl)imidazo [ 1 ,2-α]pyridin-6-y l]methylidene} - 1 ,3-thiazolidine-2,4-dione,

(5Z)-5- {[3-(hydroxyacetyl)imidazo[ 1 ,2-α]pyridin-6-yl]methylidene} -1,3- thiazolidine-2,4-dione,

(5Z)-5- {[3 -(5 -methyl- 1 ,2,4-oxadiazol-3-yl)imidazo[ 1 ,2-α]pyridin-6- yljmethylidene} - 1 ,3-thiazolidine-2,4-dione,

(5Z)-5-[(3-bromoimidazo[l,2-α]pyridin-6-yl)methylidene]- l,3- thiazolidine-2,4-dione,

(5Z)-5-[(3-fluoroimidazo[ 1 ,2-α]pyridin-6-yl)methylidene]- 1 ,3- thiazolidine-2,4-dione,

(5Z)-5-[(3-chloroimidazo[l,2-α]pyridin-6-yl)methylidene] -l,3- thiazolidine-2,4-dione,

(5Z)-5-{[3-(4-isoquinolinyl)imidazo[l,2-α]pyridin-6-yl]m ethylidene}-l,3- thiazolidine-2,4-dione,

(5Z)-5-{[3-(4-isoquinolinyl)imidazo[l,2-α]pyridin-6-yl]m ethylidene}-l,3- thiazolidine-2,4-dione,

(5Z)-5- {[3-(3-pyridinyl)imidazo[ 1 ,2-α]pyridin-6-yl]methylidene} -1,3- thiazolidine-2,4-dione,

ethyl 5- {6-[(Z)-(2,4-dioxo- 1 ,3-thiazolidin-5-ylidene)methyl]imidazo[ 1 ,2- α]pyridin-3-yl}-3-pyridinecarboxylate,

5Z)-5-{[3-(6-methyl-2-pyridinyl)imidazo[l,2-α]pyridin-6- yljmethylidene} - 1 ,3-thiazolidine-2,4-dione,

(5Z)-5-[(3-{[(l,l-dioxido-l-benzothien-6-yl)amino]methyl} imidazo[l,2- α]pyridin-6-yl)methylidene]-l,3-thiazolidine-2,4-dione,

(5 Z)S- { [3 -( { [3 -(methylsulfonyl)phenyl] amino } methyl)imidazo [ 1 ,2- α]pyridin-6-yl]methylidene}-l ,3-thiazolidine-2,4-dione, and

(5Z)-5 - [(3 - { [4-( {3 - [(trifluoromethyl)sulfonyl]phenyl} amino)- 1 - piperidinyl]methyl}imidazo[l,2-α]pyridin-6-yl)methylidene]- l,3- thiazolidine-2,4-dione; and a pharmaceutically acceptable salt thereof.

This invention also relates to a method of treating cancer, which comprises co-administering to a subject in need thereof an effective amount of a compound of Formula (I), and/or a pharmaceutically acceptable salt thereof; and at least one antineoplastic agent such as one selected from the group consisting of: anti-microtubule agents, plantinum coordination complexes, alkylating agents, antibiotic agents, topoisomerase II inhibitors, antimetabolites, topoisomerase I hinibitors, hormones and hormonal anlogues, signal transduction pathway inhibitors, non-receptor tyrosine kinase angiogenesis inhibitors, immunotherapeutic agents, proapoptotic agents, and cell cycle signaling inhibitors.

This invention also relates to a method of treating cancer, which comprises co-administering to a subject in need thereof an effective amount of a compound of Formula (I), and/or a pharmaceutically acceptable salt thereof; and at least one signal transduction pathway inhibitor such as one selected from the group consisting of: receptor tyrosine kinase inhibitor, non-receptor tyrosine kinase inhibitor, SH2/SH3 domain blocker, serine/threonine kinase inhibitor, phosphotidyl inositol-3 kinase inhibitor, myo-inositol singaling inhibitor, and Ras oncogene inhibitor.

As used herein, the term "effective amount" means that amount of a drug or pharmaceutical agent that will elicit the biological or medical response of a tissue, system, animal or human that is being sought, for instance, by a researcher or clinician. Furthermore, the term "therapeutically effective amount" means any amount which, as compared to a corresponding subject who has not received such amount, results in improved treatment, healing, prevention, or amelioration of a disease, disorder, or side effect, or a decrease in the rate of advancement of a disease or disorder. The term also includes within its scope amounts effective to enhance normal physiological function.

Compounds of Formula (I) are included in the pharmaceutical compositions of the invention. Definitions

By the term "substituted amino" as used herein, is meant -NR30R40 wherein each R30 and R40 is independently selected from a group including hydrogen, Cl- βalkyl, acyl, C3-C7cycloalkyl, wherein at least one of R30 and R40 is not hydrogen.

By the term "acyl" as used herein, unless otherwise defined, is meant

-C(O)(alkyl), -C(O)(cycloalkyl).

By the term "aryl" as used herein, unless otherwise defined, is meant aromatic, hydrocarbon, ring system. The ring system may be monocyclic or fused polycyclic (e.g. bicyclic, tricyclic, etc.). In various embodiments, the monocyclic aryl ring is C5-C10, or C5-C7, or C5-C6, where these carbon numbers refer to the number of carbon atoms that form the ring system. A C6 ring system, i.e. a phenyl ring is a suitable aryl group. In various embodiments, the polycyclic ring is a bicyclic aryl group, where suitable bicyclic aryl groups are C8-C12, or C9-C10. A naphthyl ring, which has 10 carbon atoms, is a suitable polycyclic aryl group.

By the term "heteroaryl" as used herein, unless otherwise defined, is meant

an aromatic ring system containing carbon(s) and at least one heteroatom. Heteroaryl may be monocyclic or polycyclic. A monocyclic heteroaryl group may have 1 to 4 heteroatoms in the ring, while a polycyclic heteroaryl may contain 1 to 10 hetero atoms. A polycyclic heteroaryl ring may contain fused, spiro or bridged ring junctions, for example, bicyclic heteroaryl is a polycyclic heteroaryl. Bicyclic heteroaryl rings may contain from 8 to 12 member atoms. Monocyclic heteroaryl rings may contain from 5 to 8 member atoms (carbons and heteroatoms). Exemplary heteroaryl groups include but are not limited to: benzofuran, benzothiophene, furan, imidazole, indole, isothiazole, oxazole, pyrazine, pyrazole, pyridazine, pyridine, pyrimidine, pyrrole, quinoline, quinazoline, quinoxaline, thiazole, and thiophene.

By the term "monocyclic heteroaryl" as used herein, unless otherwise defined, is meant a monocyclic heteroaryl ring containing 1-5 carbon atoms and 1-4 hetero atoms.

By the term "alkylcarboxy" as used herein, unless otherwise defined, is meant -(CH ₂ ) _n COOR ₈ o, wherein R80 is hydrogen or C 1 -Cβalkyl, n is 0-6.

By the term "alkoxy" as used herein is meant -O(alkyl) including -OCH3, - OCH ₂ CH ₃ and -OC(CH3)3 where alkyl is as described herein.

By the term "alkylthio" as used herein is meant -S(alkyl) including -SCH3, - SCH ₂ CH ₃ where where alkyl is as described herein. The term "cycloalkyl" as used herein unless otherwise defined, is meant a nonaromatic, unsaturated or saturated, cyclic or polycyclic Cβ-C^-

Examples of cycloalkyl and substituted cycloalkyl substituents as used herein include: cyclohexyl, aminocyclohexyl, cyclobutyl, aminocyclobutyl, 4- hydroxy-cyclohexyl, 2-ethylcyclohexyl, propyl4-methoxycyclohexyl, 4- methoxycyclohexyl, 4-carboxycyclohexyl, cyclopropyl, aminocyclopentyl, and cyclopentyl.

By the term "heterocycloalkyl" as used herein is meant a non-aromatic, unsaturated or saturated, monocyclic or polycyclic, heterocyclic ring containing at least one carbon and at least one heteroatom. Exemplary monocyclic heterocyclic rings include: piperidine, piperazine, pyrrolidine, and morpholine. Exemplary polycyclic heterocyclic rings include quinuclidine.

By the term "substituted" as used herein, unless otherwise defined, is meant that the subject chemical moiety has one to five substituents, suitably from one to three, selected from the group consisting of: hydrogen, halogen, Cl -Cβalkyl, amino, trifluoromethyl, -(CH ₂ ) _n COOH, C3-C7cycloalkyl, substituted amino, aryl, heteroaryl, arylalkyl, arylcycloalkyl, heteroarylalkyl, heterocycloalkyl, cyano, hydroxyl, alkoxy, alkylthio, aryloxy, acyloxy, acyl, acylamino, arylamino, nitro,

oxo, -CO ₂ R ₅₀ , -SO ₂ R ₇ O ₅ -NR ₅ OSO ₂ R ₇ C NR ₅ OC(O)R ₇₅ and -CONR ₅₅ R ₆₀ , wherein R50 and R55 are each independently selected from: hydrogen, alkyl, and C3- C7cycloalkyl; R55 and R60 can optionally form a heterocycloalkyl ring; n is 0 to 6; R75 is selected from a group consisting of: Cl-C6alkyl, aryl, substituted aryl, heteroaryl, substituted heteraryl, amino, substituted amino, arylamino, Cl- Cβheterocycloalkyl, substituted Cl-C6heterocycloalkyl; each R60 and R70 is independently selected from a group consisting of: Cl-C6alkyl, C3-C7cycloalkyl, substituted Cl-C6heterocycloalkyl, Cl-C6heterocycloalkyl, halogen, amino, substituted amino, arylamino, trifluoromethyl, cyano, hydroxyl, alkoxy, oxo, - (CH ₂ ) _n COOH, aryl optionally fused with a fϊve-membered ring or substituted with one to five groups selected from a group consisting of: Cl-C6alkyl, C3- C7cycloalkyl, halogen, amino, substituted amino, trifluoromethyl, cyano, hydroxyl, alkoxy, oxo, or -(CH ₂ ) _n COOH, or heteroaryl optionally fused with a fϊve-membered ring or substituted with one to five groups selected from a group consisting of: Cl- Cβalkyl, C3-C7cycloalkyl, halogen, amino, trifluoromethyl, cyano, hydroxyl, alkoxy, oxo, or -(CH ₂ ) _n COOH.

By the term "substituted", when referred in the definition of R60, R70, R75, "arylamino", and "aryloxy", is meant that the subject chemical moiety has one to five substituents, suitably from one to three, selected from the group consisting of: hydrogen, Cl -Cβalkyl, halogen, trifluoromethyl, -(CH ₂ ) _n COOH, amino, substituted amino, cyano, hydroxyl, alkoxy, alkylthio, aryloxy, acyloxy, acyl, acylamino, and nitro, n is 0-6.

By the term "acyloxy" as used herein is meant -OC(O)alkyl where alkyl is as described herein. Examples of acyloxy substituents as used herein include: - OC(O)CH ₃ , -OC(O)CH(CH ₃ ) ₂ and -OC(O)(CH ₂ )3CH ₃ .

By the term "acylamino" as used herein is meant -N(H)C(O)alkyl, - N(H)C(O)(cycloalkyl) where alkyl is as described herein. Examples of N- acylamino substituents as used herein include: -N(H)C(O)CH ₃ , -N(H)C(O)CH(CH ₃ ) ₂ and -N(H)C(O)(CH ₂ ) ₃ CH ₃ . By the term "aryloxy" as used herein is meant -O(aryl), -O(substituted aryl),

-O(heteroaryl) or -O(substituted heteroaryl).

By the term "arylamino" as used herein is meant -NRgo(aryl), - NR ₈ o(substituted aryl), -NRgo(heteroaryl) or -NR ₈ o(substituted heteroaryl), wherein R80 is H, Cl-6alkyl or C3-C7cycloalkyl. By the term "heteroatom" as used herein is meant oxygen, nitrogen or sulfur.

By the term "halogen" as used herein is meant a substituent selected from bromide, iodide, chloride and fluoride.

By the term "alkyl" and derivatives thereof and in all carbon chains as used herein, including alkyl chains defined by the term "-(CH2) _n ", "-(CH2) _m " and the like, is meant a linear or branched, substituted or unsubstituted, saturated or unsaturated hydrocarbon chain, and unless otherwise defined, the carbon chain will contain from 1 to 12 carbon atoms; when an alkyl group is substituted, the substituent is selected from a group consisting of: halogen, trifluoromethyl, alkylcarboxy, amino, substituted amino, cyano, hydroxyl, alkoxy, alkylthio, aryloxy, acyloxy, acyl, acylamino, and nitro, n is 0-6..

Examples of alkyl and substituted alkyl substituents as used herein include:

-CH ₃ , -CH ₂ -CH ₃ , -CH ₂ -CH ₂ -CH ₃ , -CH(CH ₃ ) ₂ , -CH ₂ -CH ₂ -C(CH ₃ ) ₃ , -CH ₂ -CF ₃ , -C≡C-C(CH ₃ ) ₃ , -C≡C-CH ₂ -OH, cyclopropylmethyl, -CH ₂ -C(CH ₃ ) ₂ -CH ₂ -NH ₂ , -C≡C-C ₆ H ₅ , -C≡C-C(CH ₃ ) ₂ -OH, -CH ₂ -CH(OH)-CH(OH)-CH(OH)-CH(OH)- CH ₂ -OH, piperidinylmethyl, methoxyphenylethyl, -C(CH ₃ ) ₃ , -(CH ₂ ) ₃ -CH ₃ , -CH ₂ - CH(CH ₃ ) ₂ , -CH(CH ₃ )-CH ₂ -CH ₃ , -CH=CH ₂ , and -C≡C-CH ₃ . By the term "treating" and derivatives thereof as used herein, is meant prophylatic and therapeutic therapy. Prophylatic therapy is meant the institution of measures to protect a person from a disease to which he or she has been, or may be, exposed. Also called preventive treatment.

By the term "co-administering" and derivatives thereof as used herein is meant either simultaneous administration or any manner of separate sequential administration of a PB kinase inhibiting compound, as described herein, and a further active ingredient or ingredients. The term further active ingredient or ingredients, as used herein, includes any compound or therapeutic agent known to or that demonstrates advantageous properties when administered to a patient in need of treatment. Suitably, if the administration is not simultaneous, the compounds are administered in a close time proximity to each other. Furthermore, it does not matter if the compounds are administered in the same dosage form, e.g. one compound may be administered topically and another compound may be administered orally. The term "compound" as used herein includes all isomers of the compound.

Examples of such isomers include: enantiomers, tautomers, rotamers.

In formulas where a "dotted" bond is drawn between two atoms, it is meant that such bond can be either single or double bond. A ring system containing such bonds can be aromatic or non-aromatic. Certain compounds described herein may contain one or more chiral atoms, or may otherwise be capable of existing as two enantiomers, or two or more

diastereoisomers. Accordingly, the compounds of this invention include mixtures of enantiomers/diastereoisomers as well as purified enantiomers/diastereoisomers or enantiomerically/diastereoisomerically enriched mixtures. Also included within the scope of the invention are the individual isomers of the compounds represented by formula I or II above as well as any wholly or partially equilibrated mixtures thereof. The present invention also covers the individual isomers of the compounds represented by the formulas above as mixtures with isomers thereof in which one or more chiral centers are inverted. Further, an example of a possible tautomer is an oxo substituent in place of a hydroxy substituent. Also, as stated above, it is understood that all tautomers and mixtures of tautomers are included within the scope of the compounds of Formula I or II.

Compounds of Formula (I) are included in the pharmaceutical compositions of the invention. Where a -COOH or -OH group is present, pharmaceutically acceptable esters can be employed, for example methyl, ethyl, pivaloyloxymethyl, and the like for -COOH, and acetate maleate and the like for -OH, and those esters known in the art for modifying solubility or hydrolysis characteristics, for use as sustained release or prodrug formulations.

It has now been found that compounds of the present invention are inhibitors of the Phosphatoinositides 3-kinases (PBKs), particularly PBKα. When the phosphatoinositides 3-kinase (PBK) enzyme is inhibited by a compound of the present invention, PBK is unable to exert its enzymatic, biological and/or pharmacological effects. The compounds of the present invention are therefore useful in the treatment of autoimmune disorders, inflammatory diseases, cardiovascular diseases, neurodegenerative diseases, allergy, asthma, pancreatitis, multiorgan failure, kidney diseases, platelet aggregation, cancer, sperm motility, transplantation rejection, graft rejection and lung injuries, particularly cancer.

Compounds according to Formula (I) are suitable for the modulation, notably the inhibition of the activity of phosphatoinositide 3-kinases (PBK), suitably phosphatoinositides 3-kinase (PBKα). Therefore the compounds of the present invention are also useful for the treatment of disorders which are mediated by PBKs. Said treatment involves the modulation - notably the inhibition or the down regulation - of the phosphatoinositides 3-kinases.

Suitably, the compounds of the present invention are used for the preparation of a medicament for the treatment of a disorder selected from multiple sclerosis, psoriasis, rheumatoid arthritis, systemic lupus erythematosis, inflammatory bowel disease, lung inflammation, thrombosis or brain infection/inflammation, such as meningitis or encephalitis, Alzheimer's disease, Huntington's disease, CNS trauma, stroke or ischemic conditions, cardiovascular diseases such as athero-sclerosis, heart hypertrophy, cardiac myocyte dysfunction, elevated blood pressure or vasoconstriction.

Suitably, the compounds of Formula (I) are useful for the treatment of autoimmune diseases or inflammatory diseases such as multiple sclerosis, psoriasis, rheumatoid arthritis, systemic lupus erythematosis, inflammatory bowel disease, lung inflammation, thrombosis or brain infection/inflammation such as meningitis or encephalitis.

Suitably, the compounds of Formula (I) are useful for the treatment of neurodegenerative diseases including multiple sclerosis, Alzheimer's disease, Huntington's disease, CNS trauma, stroke or ischemic conditions.

Suitably, the compounds of Formula (I) are useful for the treatment of cardiovascular diseases such as atherosclerosis, heart hypertrophy, cardiac myocyte dysfunction, elevated blood pressure or vasoconstriction.

Suitably, the compounds of Formula (I) are useful for the treatment of chronic obstructive pulmonary disease, anaphylactic shock fibrosis, psoriasis, allergic diseases, asthma, stroke, ischemic conditions, ischemia-reperfusion, platelets aggregation/activation, skeletal muscle atrophy/hypertrophy, leukocyte recruitment in cancer tissue, angiogenesis, invasion metastasis, in particular melanoma, Karposi's sarcoma, acute and chronic bacterial and virual infections, sepsis, transplantation rejection, graft rejection, glomerulo sclerosis, glomerulo nephritis, progressive renal fibrosis, endothelial and epithelial injuries in the lung,

and lung airway inflammation.

Because the pharmaceutically active compounds of the present invention are active as PB kinase inhibitors, particularly the compounds that inhibit PBKα, either selectively or in conjunction with one or more of PBKδ, PBKβ, and/or PBKγ, they exhibit therapeutic utility in treating cancer.

Suitably, the invention relates to a method of treating cancer in a mammal, including a human, wherein the cancer is selected from: brain (gliomas), glioblastomas, leukemias, Bannayan-Zonana syndrome, Cowden disease, Lhermitte- Duclos disease, breast, inflammatory breast cancer, Wilm's tumor, Ewing's sarcoma, Rhabdomyosarcoma, ependymoma, medulloblastoma, colon, head and neck, kidney, lung, liver, melanoma, ovarian, pancreatic, prostate, sarcoma, osteosarcoma, giant cell tumor of bone and thyroid. Suitably, the invention relates to a method of treating cancer in a mammal, including a human, wherein the cancer is selected from: Lymphoblastic T cell leukemia, Chronic myelogenous leukemia, Chronic lymphocytic leukemia, Hairy- cell leukemia, acute lymphoblastic leukemia, acute myelogenous leukemia, Chronic neutrophilic leukemia, Acute lymphoblastic T cell leukemia, Plasmacytoma, Immunoblastic large cell leukemia, Mantle cell leukemia, Multiple myeloma

Megakaryoblastic leukemia, multiple myeloma, Acute megakaryocytic leukemia, promyelocytic leukemia and Erythroleukemia.

Suitably, the invention relates to a method of treating cancer in a mammal, including a human, wherein the cancer is selected from: malignant lymphoma, hodgkins lymphoma, non-hodgkins lymphoma, lymphoblastic T cell lymphoma, Burkitt's lymphoma and follicular lymphoma.

Suitably, the invention relates to a method of treating cancer in a mammal, including a human, wherein the cancer is selected from: neuroblastoma, bladder cancer, urothelial cancer, lung cancer, vulval cancer, cervical cancer, endometrial cancer, renal cancer, mesothelioma, esophageal cancer, salivary gland cancer, hepatocellular cancer, gastric cancer, nasopharangeal cancer, buccal cancer, cancer of the mouth, GIST (gastrointestinal stromal tumor) and testicular cancer.

When a compound of Formula (I) is administered for the treatment of cancer, the term "co-administering" and derivatives thereof as used herein is meant either simultaneous administration or any manner of separate sequential administration of

a PB kinase inhibiting compound, as described herein, and a further active ingredient or ingredients, known to be useful in the treatment of cancer, including chemotherapy and radiation treatment. The term further active ingredient or ingredients, as used herein, includes any compound or therapeutic agent known to or that demonstrates advantageous properties when administered to a patient in need of treatment for cancer. Preferably, if the administration is not simultaneous, the compounds are administered in a close time proximity to each other. Furthermore, it does not matter if the compounds are administered in the same dosage form, e.g. one compound may be administered topically and another compound may be administered orally.

Typically, any anti-neoplastic agent that has activity versus a susceptible tumor being treated may be co-administered in the treatment of cancer in the present invention. Examples of such agents can be found in Cancer Principles and Practice of Oncology by V. T. Devita and S. Hellman (editors), 6 ^th edition (February 15, 2001), Lippincott Williams & Wilkins Publishers. 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. Typical antineoplastic agents useful in the present invention include, but are not limited to, anti- microtubule 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; non-receptor tyrosine kinase angiogenesis inhibitors; immunotherapeutic agents; proapoptotic agents; and cell cycle signaling inhibitors.

Examples of a further active ingredient or ingredients (anti-neoplastic agent) for use in combination or co-administered with the presently invented AKT inhibiting compounds are chemotherapeutic agents.

Anti-microtubule or anti-mitotic agents are phase specific agents 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. Diterpenoids, which are derived from natural sources, are phase specific anti

-cancer agents that operate at the G ₂ ZM phases of the cell cycle. It is believed that

the diterpenoids stabilize the β-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, paclitaxel and its analog docetaxel. Paclitaxel, 5β,20-epoxy-l,2α,4,7β,10β,13α-hexa-hydroxytax-l l-en-9-one

4,10-diacetate 2-benzoate 13-ester with (2R,3S)-N-benzoyl-3-phenylisoserine; is a natural diterpene product isolated from the Pacific yew tree Taxus brevifolia and is commercially available as an injectable solution TAXOL®. It is a member of the taxane family of terpenes. It was first isolated in 1971 by Wani et al. J. Am. Chem, Soc, 93:2325. 1971), who characterized its structure by chemical and X-ray crystallographic methods. One mechanism for its activity relates to paclitaxel's capacity to bind tubulin, thereby inhibiting cancer cell growth. Schiff et al., Proc. Natl, Acad, Sci. USA, 77:1561-1565 (1980); Schiff et al., Nature, 277:665-667 (1979); Kumar, J. Biol, Chem, 256: 10435-10441 (1981). For a review of synthesis and anticancer activity of some paclitaxel derivatives see: D. G. I. Kingston et al., Studies in Organic Chemistry vol. 26, entitled "New trends in Natural Products Chemistry 1986", Attaur-Rahman, P.W. Le Quesne, Eds. (Elsevier, Amsterdam, 1986) pp 219-235.

Paclitaxel has been approved for clinical use in the treatment of refractory ovarian cancer in the United States (Markman et al., Yale Journal of Biology and Medicine, 64:583, 1991; McGuire et al., Ann. Intern, Med., 111 :273,1989) and for the treatment of breast cancer (Holmes et al., J. Nat. Cancer Inst., 83:1797,1991.) It is a potential candidate for treatment of neoplasms in the skin (Einzig et. al., Proc. Am. Soc. Clin. Oncol., 20:46) and head and neck carcinomas (Forastire et. al., Sem. Oncol., 20:56, 1990). The compound also shows potential for the treatment of polycystic kidney disease (Woo et. al., Nature, 368:750. 1994), lung cancer and malaria. Treatment of patients with paclitaxel results in bone marrow suppression (multiple cell lineages, Ignoff, R.J. et. al, Cancer Chemotherapy Pocket Guide. _! 1998) related to the duration of dosing above a threshold concentration (5OnM) (Kearns, CM. et. al., Seminars in Oncology, 3(6) p.16-23, 1995).

Docetaxel, (2R,3S)- N-carboxy-3-phenylisoserine,N-te/t-butyl ester, 13-ester with 5β-20-epoxy-l,2α,4,7β,10β,13α-hexahydroxytax-l l-en-9-one 4-acetate 2- benzoate, trihydrate; is commercially available as an injectable solution as TAXOTERE®. Docetaxel is indicated for the treatment of breast cancer. 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. The dose limiting toxicity of docetaxel is neutropenia.

Vinca alkaloids are phase specific anti-neoplastic agents derived from the periwinkle plant. Vinca alkaloids 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, and vinorelbine.

Vinblastine, vincaleukoblastine sulfate, is commercially available as VELB AN® as an injectable solution. Although, it has possible indication as a second line therapy of various solid tumors, it is primarily indicated in the treatment of testicular cancer and various lymphomas including Hodgkin's Disease; and lymphocytic and histiocytic lymphomas. Myelosuppression is the dose limiting side effect of vinblastine.

Vincristine, vincaleukoblastine, 22-oxo-, sulfate, is commercially available as ONCOVIN® as an injectable solution. Vincristine is indicated for the treatment of acute leukemias and has also found use in treatment regimens for Hodgkin's and non-Hodgkin's malignant lymphomas. Alopecia and neurologic effects are the most common side effect of vincristine and to a lesser extent myelosupression and gastrointestinal mucositis effects occur.

Vinorelbine, 3',4'-didehydro -4'-deoxy-C'-norvincaleukoblastine [R- (R*,R*)-2,3-dihydroxybutanedioate (1 :2)(salt)], commercially available as an injectable solution of vinorelbine tartrate (NA VELB INE®), is a semisynthetic vinca alkaloid. Vinorelbine is indicated as a single agent or in combination with other chemotherapeutic agents, such as cisplatin, in the treatment of various solid tumors, particularly non-small cell lung, advanced breast, and hormone refractory prostate cancers. Myelosuppression is the most common dose limiting side effect of vinorelbine.

Platinum coordination complexes are non-phase specific anti-cancer agents, which are interactive with DNA. The platinum complexes enter tumor cells, undergo, aquation and form intra- and interstrand crosslinks with DNA causing adverse biological effects to the tumor. Examples of platinum coordination complexes include, but are not limited to, cisplatin and carboplatin.

Cisplatin, cis-diamminedichloroplatinum, is commercially available as PLATINOL® as an injectable solution. Cisplatin is primarily indicated in the treatment of metastatic testicular and ovarian cancer and advanced bladder cancer. The primary dose limiting side effects of cisplatin are nephrotoxicity, which may be controlled by hydration and diuresis, and ototoxicity.

Carboplatin, platinum, diammine [l,l-cyclobutane-dicarboxylate(2-)-O,O'],

is commercially available as PARAPLATIN® as an injectable solution. Carboplatin is primarily indicated in the first and second line treatment of advanced ovarian carcinoma. Bone marrow suppression is the dose limiting toxicity of carboplatin.

Alkylating agents are non-phase anti-cancer specific agents and 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, nitrogen mustards such as cyclophosphamide, melphalan, and chlorambucil; alkyl sulfonates such as busulfan; nitrosoureas such as carmustine; and triazenes such as dacarbazine.

Cyclophosphamide, 2-[bis(2-chloroethyl)amino]tetrahydro-2H-l,3,2- oxazaphosphorine 2-oxide monohydrate, is commercially available as an injectable solution or tablets as CYTOXAN®. Cyclophosphamide is indicated as a single agent or in combination with other chemotherapeutic agents, in the treatment of malignant lymphomas, multiple myeloma, and leukemias. Alopecia, nausea, vomiting and leukopenia are the most common dose limiting side effects of cyclophosphamide.

Melphalan, 4-[bis(2-chloroethyl)amino]-L-phenylalanine, is commercially available as an injectable solution or tablets as ALKERAN®. Melphalan is indicated for the palliative treatment of multiple myeloma and non-resectable epithelial carcinoma of the ovary. Bone marrow suppression is the most common dose limiting side effect of melphalan.

Chlorambucil, 4-[bis(2-chloroethyl)amino]benzenebutanoic acid, is commercially available as LEUKERAN® tablets. Chlorambucil is indicated for the palliative treatment of chronic lymphatic leukemia, and malignant lymphomas such as lymphosarcoma, giant follicular lymphoma, and Hodgkin's disease. Bone marrow suppression is the most common dose limiting side effect of chlorambucil. Busulfan, 1,4-butanediol dimethanesulfonate, is commercially available as MYLERAN® TABLETS. Busulfan is indicated for the palliative treatment of chronic myelogenous leukemia. Bone marrow suppression is the most common dose limiting side effects of busulfan.

Carmustine, 1, 3 -[bis(2-chloroethyl)-l -nitrosourea, is commercially available as single vials of lyophilized material as BiCNU®. Carmustine is indicated for the palliative treatment as a single agent or in combination with other agents for brain tumors, multiple myeloma, Hodgkin's disease, and non-Hodgkin's lymphomas. Delayed myelosuppression is the most common dose limiting side effects of

carmustine.

Dacarbazine, 5-(3,3-dimethyl-l-triazeno)-imidazole-4-carboxamide, is commercially available as single vials of material as DTIC-Dome®. Dacarbazine is indicated for the treatment of metastatic malignant melanoma and in combination with other agents for the second line treatment of Hodgkin's Disease. Nausea, vomiting, and anorexia are the most common dose limiting side effects of dacarbazine.

Antibiotic anti-neoplasties are non-phase specific agents, which bind or intercalate with DNA. Typically, such action results in stable DNA complexes or strand breakage, which disrupts ordinary function of the nucleic acids leading to cell death. Examples of antibiotic anti-neoplastic agents include, but are not limited to, actinomycins such as dactinomycin, anthrocyclins such as daunorubicin and doxorubicin; and bleomycins.

Dactinomycin, also know as Actinomycin D, is commercially available in injectable form as COSMEGEN®. Dactinomycin is indicated for the treatment of

WiIm' s tumor and rhabdomyosarcoma. Nausea, vomiting, and anorexia are the most common dose limiting side effects of dactinomycin.

Daunorubicin, (8S-cis-)-8-acetyl-10-[(3-amino-2,3,6-trideoxy-α-L-lyxo- hexopyranosyl)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®. Daunorubicin is indicated for remission induction in the treatment of acute nonlymphocytic leukemia and advanced HIV associated Kaposi's sarcoma. Myelosuppression is the most common dose limiting side effect of daunorubicin. Doxorubicin, (8S, 10S)-10-[(3-amino-2,3,6-trideoxy-α-L-lyxo- hexopyranosyl)oxy]-8-glycoloyl, 7,8,9, 10-tetrahydro-6,8, 11 -trihydroxy- 1 -methoxy-

5,12 naphthacenedione hydrochloride, is commercially available as an injectable form as RUBEX® or ADRI AMYCIN RDF®. Doxorubicin is primarily indicated for the treatment of acute lymphoblastic leukemia and acute myeloblasts leukemia, but is also a useful component in the treatment of some solid tumors and lymphomas. Myelosuppression is the most common dose limiting side effect of doxorubicin.

Bleomycin, a mixture of cytotoxic glycopeptide antibiotics isolated from a strain of Streptomyces verticillus, is commercially available as BLENOXANE®. Bleomycin is indicated as a palliative treatment, as a single agent or in combination with other agents, of squamous cell carcinoma, lymphomas, and testicular carcinomas. Pulmonary and cutaneous toxicities are the most common dose limiting

side effects of bleomycin.

Topoisomerase II inhibitors include, but are not limited to, epipodophyllotoxins .

Epipodophyllotoxins are phase specific anti-neoplastic agents derived from the mandrake plant. Epipodophyllotoxins typically affect cells in the S and G ₂ 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 and teniposide. Etoposide, 4'-demethyl-epipodophyllotoxin 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. Etoposide is indicated as a single agent or in combination with other chemotherapy agents in the treatment of testicular and non-small cell lung cancers. Myelosuppression is the most common side effect of etoposide. The incidence of leucopenia tends to be more severe than thrombocytopenia.

Teniposide, 4'-demethyl-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. Teniposide is indicated as a single agent or in combination with other chemotherapy agents in the treatment of acute leukemia in children. Myelosuppression is the most common dose limiting side effect of teniposide. Teniposide can induce both leucopenia and thrombocytopenia.

Antimetabolite neoplastic agents are phase specific anti-neoplastic agents that 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. Examples of antimetabolite anti-neoplastic agents include, but are not limited to, fluorouracil, methotrexate, cytarabine, mecaptopurine, thioguanine, and gemcitabine. 5 -fluorouracil, 5-fluoro-2,4- (1H,3H) pyrimidinedione, is commercially available as fluorouracil. Administration of 5 -fluorouracil leads to inhibition of thymidylate synthesis and is also incorporated into both RNA and DNA. The result typically is cell death. 5 -fluorouracil is indicated as a single agent or in combination with other chemotherapy agents in the treatment of carcinomas of the breast, colon, rectum, stomach and pancreas. Myelosuppression and mucositis are dose limiting side effects of 5-fluorouracil. Other fluoropyrimidine analogs include 5-fluoro deoxyuridine (floxuridine) and 5-fluorodeoxyuridine monophosphate.

Cytarabine, 4-amino-l-β-D-arabinofuranosyl-2 (lH)-pyrimidinone, is commercially available as CYTOSAR-U® and is commonly known as Ara-C. It is believed that cytarabine exhibits cell phase specificity at S-phase by inhibiting DNA chain elongation by terminal incorporation of cytarabine into the growing DNA chain. Cytarabine is indicated as a single agent or in combination with other chemotherapy agents in the treatment of acute leukemia. Other cytidine analogs include 5-azacytidine and 2', 2 '-difluorodeoxy cytidine (gemcitabine). Cytarabine induces leucopenia, thrombocytopenia, and mucositis.

Mercaptopurine, l,7-dihydro-6H-purine-6-thione monohydrate, is commercially available as PURINE THOL®. Mercaptopurine exhibits cell phase specificity at S-phase by inhibiting DNA synthesis by an as of yet unspecified mechanism. Mercaptopurine is indicated as a single agent or in combination with other chemotherapy agents in the treatment of acute leukemia. Myelosuppression and gastrointestinal mucositis are expected side effects of mercaptopurine at high doses. A useful mercaptopurine analog is azathioprine.

Thioguanine, 2-amino-l,7-dihydro-6H-purine-6-thione, is commercially available as TABLOID®. Thioguanine exhibits cell phase specificity at S-phase by inhibiting DNA synthesis by an as of yet unspecified mechanism. Thioguanine is indicated as a single agent or in combination with other chemotherapy agents in the treatment of acute leukemia. Myelosuppression, including leucopenia, thrombocytopenia, and anemia, is the most common dose limiting side effect of thioguanine administration. However, gastrointestinal side effects occur and can be dose limiting. Other purine analogs include pentostatin, erythrohydroxynonyladenine, fludarabine phosphate, and cladribine. Gemcitabine, 2'-deoxy-2', 2'-difluorocytidine monohydrochloride (β- isomer), is commercially available as GEMZAR®. Gemcitabine exhibits cell phase specificity at S-phase and by blocking progression of cells through the Gl/S boundary. Gemcitabine is indicated in combination with cisplatin in the treatment of locally advanced non-small cell lung cancer and alone in the treatment of locally advanced pancreatic cancer. Myelosuppression, including leucopenia, thrombocytopenia, and anemia, is the most common dose limiting side effect of gemcitabine administration.

Methotrexate, N- [4 [ [(2,4-diamino-6-pteridinyl) methyljmethylamino] benzoyl]-L-glutamic acid, is commercially available as methotrexate sodium. Methotrexate exhibits cell phase effects specifically at S-phase by inhibiting DNA synthesis, repair and/or replication through the inhibition of dyhydro folic acid reductase which is required for synthesis of purine nucleotides and thymidylate.

Methotrexate is indicated as a single agent or in combination with other chemotherapy agents in the treatment of choriocarcinoma, meningeal leukemia, non- Hodgkin's lymphoma, and carcinomas of the breast, head, neck, ovary and bladder. Myelosuppression (leucopenia, thrombocytopenia, and anemia) and mucositis are expected side effect of methotrexate administration.

Camptothecins, including, camptothecin and camptothecin derivatives are available or under development as Topoisomerase I inhibitors. Camptothecins cytotoxic activity is believed to be related to its Topoisomerase I inhibitory activity. Examples of camptothecins include, but are not limited to irinotecan, topotecan, and the various optical forms of 7-(4-methylpiperazino-methylene)- 10,11- ethylenedioxy-20-camptothecin described below.

Irinotecan HCl, (4S)-4,1 l-diethyl-4-hydroxy-9-[(4-piperidinopiperidino) carbonyloxy]-lH-pyrano[3',4',6,7]indolizino[l,2-b]quinoline- 3,14(4H,12H)-dione hydrochloride, is commercially available as the injectable solution CAMPTOSAR®. Irinotecan is a derivative of camptothecin which binds, along with its active metabolite SN-38, to the topoisomerase I - DNA complex. It is believed that cytotoxicity occurs as a result of irreparable double strand breaks caused by interaction of the topoisomerase I : DNA : irintecan or SN-38 ternary complex with replication enzymes. Irinotecan is indicated for treatment of metastatic cancer of the colon or rectum. The dose limiting side effects of irinotecan HCl are myelosuppression, including neutropenia, and GI effects, including diarrhea.

Topotecan HCl, (S)-10-[(dimethylamino)methyl]-4-ethyl-4,9-dihydroxy-lH- pyrano[3',4',6,7]indolizino[l,2-b]quinoline-3,14-(4H,12H)-di one monohydrochloride, is commercially available as the injectable solution HYCAMTIN®. Topotecan is a derivative of camptothecin which binds to the topoisomerase I - DNA complex and prevents religation of singles strand breaks caused by Topoisomerase I in response to torsional strain of the DNA molecule. Topotecan is indicated for second line treatment of metastatic carcinoma of the ovary and small cell lung cancer. The dose limiting side effect of topotecan HCl is myelosuppression, primarily neutropenia.

Also of interest, is the camptothecin derivative of formula A following, currently under development, including the racemic mixture (R, S) form as well as the R and S enantiomers:

known by the chemical name "7-(4-methylpiperazino-methylene)-10,l 1- ethylenedioxy-20(R,S)-camptothecin (racemic mixture) or "7-(4-methylpiperazino- methylene)- 10,11 -ethylenedioxy-20(R)-camptothecin (R enantiomer) or "7-(4- methylpiperazino-methylene)- 10,11 -ethylenedioxy-20(S)-camptothecin (S enantiomer). Such compound as well as related compounds are described, including methods of making, 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. 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, adrenocorticosteroids such as prednisone and prednisolone which are useful in the treatment of malignant lymphoma and acute leukemia in children; aminoglutethimide and other aromatase inhibitors such as anastrozole, letrazole, vorazole, and exemestane useful in the treatment of adrenocortical carcinoma and hormone dependent breast carcinoma containing estrogen receptors; progestrins such as megestrol acetate useful in the treatment of hormone dependent breast cancer and endometrial carcinoma; estrogens, androgens, and anti-androgens such as flutamide, nilutamide, bicalutamide, cyproterone acetate and 5α-reductases such as finasteride and dutasteride, useful in the treatment of prostatic carcinoma and benign prostatic hypertrophy; anti-estrogens such as tamoxifen, toremifene, raloxifene, droloxifene, 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, useful in the treatment of hormone dependent breast carcinoma and other susceptible cancers; and gonadotropin-releasing hormone (GnRH) and analogues thereof which stimulate the release of leutinizing hormone (LH) and/or follicle stimulating hormone (FSH) for the treatment prostatic carcinoma, for instance, LHRH agonists and antagagonists such as goserelin acetate and luprolide.

Signal transduction pathway inhibitors are those inhibitors, which block or inhibit a chemical process which evokes an intracellular change. As used herein this change is cell proliferation or differentiation. Signal tranduction inhibitors useful in the present invention include inhibitors of receptor tyrosine kinases, non-receptor tyrosine kinases, SH2/SH3domain blockers, serine/threonine kinases, phosphotidyl inositol-3 kinases, myo-inositol signaling, and Ras oncogenes.

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 generally termed growth factor receptors. Inappropriate or uncontrolled activation of many of these kinases, i.e. aberrant kinase growth factor receptor activity, 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, erbB4, 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 DDT VoI 2, No. 2 February 1997; 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.

Tyrosine kinases, which are not growth factor receptor kinases are termed non-receptor tyrosine kinases. Non-receptor tyrosine kinases for use 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., (1999) Journal of Hematotherapy and Stem Cell Research 8 (5): 465 - 80; and Bolen, J.B., Brugge, J.S., (1997) Annual review of Immunology. 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 (She, Crk, Nek, Grb2) and Ras-GAP. SH2/SH3 domains as targets for anti-cancer drugs are discussed in Smithgall, T. E. (1995), Journal of Pharmacological and Toxicological Methods. 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., (1999), Journal of Biochemistry. 126 (5) 799-803; Brodt, P, Samani, A., and Navab, R. (2000), Biochemical Pharmacology, 60. 1101-1107; Massague, J., Weis-Garcia, F. (1996) Cancer Surveys. 27:41-64; Philip, P.A., and Harris, A.L. (1995), Cancer Treatment and Research. 78: 3-27, Lackey, K. et al

Bioorganic and Medicinal Chemistry Letters, (10), 2000, 223-226; U.S. Patent No. 6,268,391; and Martinez-Iacaci, L., 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 may also be useful in the present invention. Such kinases are discussed in Abraham, R.T. (1996), Current Opinion in Immunology. 8 (3) 412-8; Canman, C.E., Lim, D.S. (1998), Oncogene 17 (25) 3301- 3308; Jackson, S.P. (1997), International Journal of Biochemistry and Cell Biology. 29 (7):935-8; and Zhong, H. et al, Cancer res, (2000) 60(6), 1541-1545.

Also of interest in the present invention are Myo-inositol signaling inhibitors such as phospho lipase 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, S.I. Matar, P. (2000), Journal of Biomedical Science. 7(4) 292-8; Ashby, M.N. (1998), Current Opinion in Lipidology. 9 (2) 99 - 102; and BioChim. Biophys. Acta, (19899) 1423(3):19-30.

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, Monoclonal Antibody Therapy for Solid Tumors, Cancer Treat. Rev., (2000), 26(4), 269-286); Herceptin ® erbB2 antibody (see Tyrosine Kinase Signalling in Breast cancer:erbB Family Receptor Tyrosine Kniases, Breast cancer Res., 2000, 2(3), 176-183); and 2CB VEGFR2 specific antibody (see Brekken, R. A. et al, Selective Inhibition of VEGFR2 Activity by a monoclonal Anti-VEGF antibody blocks tumor growth in mice, Cancer Res. (2000) 60, 5117-5124).

Non-receptor kinase angiogenesis inhibitors may also be useful 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. Accordingly, non-receptor tyrosine kinase inhibitors may be used in combination with the compounds 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 (alpha _v beta ₃ ) that will inhibit angiogenesis; endostatin and angiostatin (non-RTK) may also prove useful in combination with the disclosed compounds. (See Bruns CJ et al (2000), Cancer Res., 60: 2926-2935; Schreiber AB, Winkler ME, and Derynck R. (1986), Science, 232: 1250-1253; Yen L et al. (2000), Oncogene 19: 3460-3469).

Agents used in immunotherapeutic regimens may also be useful in combination with the compounds of formula (I). There are a number of immunologic strategies to generate an immune response. These strategies are generally in the realm of tumor vaccinations. The efficacy of immunologic approaches may be greatly enhanced through combined inhibition of signaling pathways using a small molecule inhibitor. Discussion of the immuno logic/tumor vaccine approach against erbB2/EGFR are found in Reilly RT et al. (2000), Cancer

Res. 60: 3569-3576; and Chen Y, Hu D, Eling DJ, Robbins J, and Kipps TJ. (1998), Cancer Res. 58: 1965-1971.

Agents used in proapoptotic 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 (i.e., mcl-1). 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 proapoptotic strategies using the antisense oligonucleotide strategy for bcl-2 are discussed in Water JS et al. (2000), J. Clin. Oncol. 18: 1812-1823; and Kitada S et al. (1994), Antisense Res. Dev. 4: 71-79.

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 signalling 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, Rosania et al, Exp. Opin. Ther. Patents (2000) 10(2):215-230.

In one embodiment, the cancer treatment method of the claimed invention includes the co -administration a compound of formula I and/or a pharmaceutically acceptable salt thereof and at least one anti-neoplastic agent, such as one selected from the group consisting of anti-microtubule agents, platinum coordination complexes, alkylating agents, antibiotic agents, topoisomerase II inhibitors, antimetabolites, topoisomerase I inhibitors, hormones and hormonal analogues, signal transduction pathway inhibitors, non-receptor tyrosine kinase angiogenesis inhibitors, immunotherapeutic agents, proapoptotic agents, and cell cycle signaling inhibitors.

Because the pharmaceutically active compounds of the present invention are active as PI3 kinase inhibitors, particularly the compounds that modulate/inhibit PI3Kα, it is useful in treating cancer. Because the pharmaceutically active compounds of the present invention are also active against one or more of PI3Kδ, PI3Kβ, and/or PI3Kγ, they exhibit therapeutic utility in treating a disease state selected from: autoimmune disorders, inflammatory diseases, cardiovascular

diseases, neurodegenerative diseases, allergy, asthma, pancreatitis, multiorgan failure, kidney diseases, platelet aggregation, sperm motility, transplantation rejection, graft rejection and lung injuries.

When a compound of Formula (I) is administered for the treatment of a disease state selected from: autoimmune disorders, inflammatory diseases, cardiovascular diseases, neurodegenerative diseases, allergy, cancer, asthma, pancreatitis, multiorgan failure, kidney diseases, platelet aggregation, sperm motility, transplantation rejection, graft rejection or lung injuries, the term "co- administering" and derivatives thereof as used herein is meant either simultaneous administration or any manner of separate sequential administration of a PB kinase inhibiting compound, as described herein, and a further active ingredient or ingredients, known to be useful in the treatment of such autoimmune disorder, cancer, inflammatory diseases, cardiovascular disease, neurodegenerative disease, allergy, asthma, pancreatitis, multiorgan failure, kidney diseases, platelet aggregation, sperm motility, transplantation rejection, graft rejection and/or lung injuries.

Biological assays PI3K alpha Leadseeker SPA Assay

Compounds of the present invention were tested according to the following assays and found as inhibitors of PB kinases, particularly PBKα. The activities (ICso) of exemplified compounds range from 1 nM to 10 μM against PBKα. Assay principle SPA imaging beads are microspheres containing scintillant which emit light in the red region of the visible spectrum. As a result, these beads are ideally suited to use with a CCD imager such as the Viewlux. The Leadseeker beads used in this system are polystyrene beads that have been coupled with polyethyleneimine. When added to the assay mixture, the beads absorb both the substrate (PIP2) and product (PIP3). Adsorbed P ³³ -PIP3 will cause an increase in signal, measured as ADUs (analog to digital units). This protocol details the use of the PEI-PS Leadseeker beads for assays using His-pl 10/p85 PBK alpha.

Assay protocol Solid compounds are typically plated with 0.1 μl of 100% DMSO in all wells (except column 6 and 18) of a 384-well, flat bottom, low volume plate (Greiner 784075). The compounds are serially diluted (3-fold in 100% DMSO) across the

plate from column 1 to column 12 and column 13 to column 24 and leave column 6 and 18 containing only DMSO to yield 1 lconcentraions for each test compound.

The assay buffer contains MOPS (pH 6.5), CHAPS, and DTT. PBK alpha and PIP2 (L-alpha-D-myo-Phosphatidylinositol 4,5-bisphosphate [PI(4,5)P2]3-O-phospho linked, D(+)-sn-l,2-di-O-octanoylglyceryl, CellSignals # 901) are mixed and incubated in the plate with compound for 30min prior to starting the reaction with the addition of P ³³ -ATP and MgCl ₂ (reagents added using Zoom). Enzyme-free wells (column 18) are typically done to determine the low control. PEI-PS Leadseeker beads in PBS/EDTA/CHAPS are added (by Multidrop) to quench the reaction, and the plates are allowed to incubate for at least one hour (typically overnight) before centrifugation. The signal is determined using a Viewlux detector and is then imported into curve fitting software (Activity Base) for construction of concentration response curves. The percent inhibition of activity was calculated relative to high controls (Cl, 0.1 μl DMSO in column 6, rows A-P)) and low controls (C2, 5 μl of 40 uM PIP2 in buffer in column 18, rows A-P) using, 100*(l-(Ul- C2)/(C1-C2)). The concentration of test compound yielding 50% inhibition was determined using the equation, y = ((Vmax*x) / (K+x)) + Y2, where "K" was equal to the IC50. The IC50 values were converted to pIC50 values, i.e., -log IC50 in Molar concentration.

Celluar assays:

DAY l

• Plate cells before noon o 1 OK cells/well in clear flat-bottomed 96-well plates if. v. 105ul) o Last four wells in last column receive media only o Place in 37degC incubator overnight

• Compound plate o Prepare in polypropylene round-bottomed 96-well plates; 8 compounds per plate, 11-pt titrations of each (3x serial dilution), DMSO in last column

(0.15% f.c. on cells) o 15ul in first well, lOul DMSO in the rest; take 5ul from first well and mix in next, continue across plate {excluding last column); seal with foil lid and place at 4degC

DAY 2

• Take out Lysis buffer inhibitors (4degC/-20degC) and compound plates (4degC), thaw on bench top; make Ix Tris wash buffer (WB) to fill reservoir on plate washer and top off bench supply (use MiIiQ), turn on centrifuge to allow it to cool

• Block MSD plates o Make 20ml 3% blocking solution/plate (600 mg blocker A in 20ml WB), add 150ul/well and incubate at RT for at least 1 hr

• Add compound (while blocking) o Add 300ul growth media (RPMI w/ Q, 10% FBS) per well (682x dil of compound) to each compound plate o Add 5ul compound dilution into each well (f.v. 11 OuI) on duplicate plates o Place in 37degC incubator for 30min • Make lysates o Prepare MSD Lysis buffer; for 10ml add 200ul protease inhibitor solution, and lOOul each of Phosphatase inhibitors I & II {Keep on ice until ready for use) o Remove plates post-incubation, aspirate media with plate washer, wash Ix with cold PBS, and add 80ul MSD Lysis buffer per well; incubate on shaker at 4degC for >30min o Spin cold at 2500rpm for lOmin; leave plates in 4degC centrifuge until ready for use

• AKT duplex assay o Wash plates (4x with 200ul/well WB in plate washer); tap plates on paper towel to blot o Add 60ul of lysates/well, incubate on shaker at RT for 1 hr o During incubation prepare detection Ab (3 ml/plate; 2 ml WB and 1 ml blocking solution w/ Ab at 1OnM); repeat wash step as above o Add 25ul of Ab/well, incubate on shaker at RT for 1 hr; repeat wash step as above o Add 150ul/well Ix Read Buffer (dilute 4x stock in ddH2O, 20ml/plate), read immediately

• Analysis o Observe all the data points at each compound concentration. o The data point from highest inhibitor concentration must be equal or greater than 70% of DMSO control. o IC50 for duplicate runs must be within 2-fold of each other (not flagged in summary template). o Y min must be greater than zero; if both mins are red flagged (>35) then compound is listed as inactive (IC50= > highest dose). If only one min is red flagged, but still <50 then call IC50 as listed. o Any data points equal or greater than 30% off the curve will not be considered. Cell Growth/Death Assay:

BT474, HCC 1954 and T-47D (human breast) were cultured in RPMI- 1640 containing 10% fetal bovine serum at 37 ⁰ C in 5% CO ₂ incubator. Cells were split into T75 flask (Falcon #353136) two to three days prior to assay set up at density which yields approximately 70-80% confluence at time of harvest for assay. Cells

were harvested using 0.25% trypsin-EDTA (Sigma #4049). Cell counts were performed on cell suspension using Trypan Blue exclusion staining. Cells were then plated in 384 well black flat bottom polystyrene (Greiner #781086) in 48 μl of culture media per well at 1,000 cells/well. All plates were placed at 5% CO ₂ , 37 ⁰ C overnight and test compounds were added the following day. One plate was treated with CellTiter-Glo (Promega #G7573) for a day 0 (t=0) measurement and read as described below. The test compounds were prepared in clear bottom polypropylene 384 well plates (Greiner#781280) with consecutive two fold dilutions. 4 μl of these dilutions were added to 105 μl culture media, after mixing the solution, 2 μl of these dilutions were added into each well of the cell plates. The final concentration of

DMSO in all wells was 0.15%. Cells were incubated at 37 ⁰ C, 5% CO ₂ for 72 hours. Following 72 hours of incubation with compounds each plate was developed and read. CellTiter-Glo reagent was added to assay plates using a volume equivalent to the cell culture volume in the wells. Plates were shaken for approximately two minutes and incubated at room temperature for approximately 30 minutes and chemiluminescent signal was read on the Analyst GT (Molecular Devices) reader. Results were expressed as a percent of the t=0 and plotted against the compound concentration. Cell growth inhibition was determined for each compound by fitting the dose response with a 4 or 6 parameter curve fit using XLfit software and determining the concentration that inhibited 50% of the cell growth (gIC50) with the Y min as the t=0 and Y max as the DMSO control. Value from wells with no cells was subtracted from all samples for background correction. Additional references:

The compounds of the present invention can also be tested to determine their inhibitory activity at PI3Kα, PI3Kδ, PI3Kβ and PI3Kγ according to the assays in the following references: For all PI3K iso forms:

1. Cloning, expression, purification, and characterization of the human Class Ia phosphoinositide 3-kinase isoforms: Meier, T.I.; Cook, J.A.; Thomas, J.E.; Radding, J.A.; Horn, C; Lingaraj, T.; Smith, M. C. Protein Expr. Purif,

2004, 35(2), 218.

2. Competitive fluorescence polarization assays for the detection of

phosphoinositide kinase and phosphatase activity: Drees, B. E.; Weipert, A.; Hudson, H.; Ferguson, C. G.; Chakravarty, L.; Prestwich, G.D. Comb. Chem. High Throughput.Screen., 2003, 6(4), 321. For PI3Kγ: WO 2005/011686 Al The pharmaceutically active compounds within the scope of this invention are useful as PB Kinase inhibitors in mammals, particularly humans, in need thereof.

The present invention therefore provides a method of treating diseases associated with PB kinase inhibition, particularly: autoimmune disorders, inflammatory diseases, cardiovascular diseases, neurodegenerative diseases, allergy, asthma, pancreatitis, multiorgan failure, kidney diseases, platelet aggregation, cancer, sperm motility, transplantation rejection, graft rejection and lung injuries and other conditions requiring PB kinase modulation/inhibition, which comprises administering an effective compound of Formula (I) or a pharmaceutically acceptable salt thereof. The compounds of Formula (I) also provide for a method of treating the above indicated disease states because of their ability to act as PB inhibitors. The drug may be administered to a patient in need thereof by any conventional route of administration, including, but not limited to, intravenous, intramuscular, oral, subcutaneous, intradermal, and parenteral. The pharmaceutically active compounds of the present invention are incorporated into convenient dosage forms such as capsules, tablets, or injectable preparations. Solid or liquid pharmaceutical carriers are employed. Solid carriers include, starch, lactose, calcium sulfate dihydrate, terra alba, sucrose, talc, gelatin, agar, pectin, acacia, magnesium stearate, and stearic acid. Liquid carriers include syrup, peanut oil, olive oil, saline, and water. Similarly, the carrier or diluent may include any prolonged release material, such as glyceryl monostearate or glyceryl distearate, alone or with a wax. The amount of solid carrier varies widely but, preferably, will be from about 25 mg to about 1 g per dosage unit. When a liquid carrier is used, the preparation will be in the form of a syrup, elixir, emulsion, soft gelatin capsule, sterile injectable liquid such as an ampoule, or an aqueous or nonaqueous liquid suspension.

The pharmaceutical preparations are made following conventional techniques of a pharmaceutical chemist involving mixing, granulating, and compressing, when necessary, for tablet forms, or mixing, filling and dissolving the ingredients, as appropriate, to give the desired oral or parenteral products.

Doses of the presently invented pharmaceutically active compounds in a

pharmaceutical dosage unit as described above will be an efficacious, nontoxic quantity preferably selected from the range of 0.001 - 100 mg/kg of active compound, preferably 0.001 - 50 mg/kg. When treating a human patient in need of a PBK inhibitor, the selected dose is administered preferably from 1-6 times daily, orally or parenterally. Preferred forms of parenteral administration include topically, rectally, transdermally, by injection and continuously by infusion. Oral dosage units for human administration preferably contain from 0.05 to 3500 mg of active compound. Oral administration, which uses lower dosages is preferred. Parenteral administration, at high dosages, however, also can be used when safe and convenient for the patient.

Optimal dosages to be administered may be readily determined by those skilled in the art, and will vary with the particular PB kinase inhibitor in use, the strength of the preparation, the mode of administration, and the advancement of the disease condition. Additional factors depending on the particular patient being treated will result in a need to adjust dosages, including patient age, weight, diet, and time of administration.

The method of this invention of inducing PB kinase inhibitory activity in mammals, including humans, comprises administering to a subject in need of such activity an effective PB kinase modulating/inhibiting amount of a pharmaceutically active compound of the present invention.

The invention also provides for the use of a compound of Formula (I) in the manufacture of a medicament for use as a PB kinase inhibitor.

The invention also provides for the use of a compound of Formula (I) in the manufacture of a medicament for use in therapy. The invention also provides for the use of a compound of Formula (I) in the manufacture of a medicament for use in treating autoimmune disorders, inflammatory diseases, cardiovascular diseases, neurodegenerative diseases, allergy, asthma, pancreatitis, multiorgan failure, kidney diseases, platelet aggregation, cancer, sperm motility, transplantation rejection, graft rejection and lung injuries. The invention also provides for a pharmaceutical composition for use as a

PB inhibitor which comprises a compound of Formula (I) and a pharmaceutically acceptable carrier.

The invention also provides for a pharmaceutical composition for use in the treatment of autoimmune disorders, inflammatory diseases, cardiovascular diseases, neurodegenerative diseases, allergy, asthma, pancreatitis, multiorgan failure, kidney diseases, platelet aggregation, cancer, sperm motility, transplantation rejection, graft rejection and lung injuries, which comprises a compound of Formula (I) and a

pharmaceutically acceptable carrier.

No unacceptable toxicological effects are expected when compounds of the invention are administered in accordance with the present invention.

In addition, the pharmaceutically active compounds of the present invention can be co-administered with further active ingredients, including compounds known to have utility when used in combination with a PB kinase inhibitor.

Without further elaboration, it is believed that one skilled in the art can, using the preceding description, utilize the present invention to its fullest extent. The following examples are, therefore, to be construed as merely illustrative and not a limitation of the scope of the present invention in any way.

Experimental Details

The compounds of the following examples are readily made according to the following scheme(s) or by analogous methods. SCHEMES:

General Scheme I:

I I I I l I V

Conditions: a) NaHCO ₃ , i-PrOH, 80 ⁰ C or cone. H ₂ SO ₄ , EtOH, 90 ⁰ C; b) CH ₂ CHBF ₃ K, PdCI ₂ (dppf)-CH ₂ CI ₂ , TEA, n-PrOH, 100 ⁰ C; c) OsO ₄ , NaIO ₄ , 2,6- lutidine, f-BuOH, dioxane, H ₂ O; d) 2,4-thiazolidinedione, piperidine, AcOH, EtOH, 90 ⁰ C or β-alanine, AcOH, 100 ⁰ C.

General Scheme II:

Conditions: a) NaHCO ₃ , i-PrOH, 80 ⁰ C; b) NIS, MeCN, rt; c) Het-B(OR) ₂ , Pd(PPh ₃ ) ₄ , K ₂ CO ₃ (aq), dioxane, 100 ⁰ C; d) CH ₂ CHBF ₃ K, PdCI ₂ (dppf)-CH ₂ CI ₂ , TEA, n-PrOH, 100 ⁰ C; e) OsO ₄ , NaIO ₄ , 2,6-lutidine, f-BuOH, dioxane, H ₂ O, rt; f) 2,4- thiazolidinedione, piperidine, AcOH, EtOH, 90 ⁰ C or β-alanine, AcOH, 100 ⁰ C.

General Scheme III:

X = Br or Cl

Conditions: a) MeOH, sealed tube, 70 ⁰ C; b) NaHCO ₃ , i-PrOH, sealed tube , 100 ⁰ C.

General Scheme IV:

Conditions: a) Pd(PPh ₃ ) ₄ , 1,4-dioxane, 100 ⁰ C, sealed tube; b) HONH ₂ -HCl, TEA, EtOH; c) Ac ₂ O, toluene, 100 ⁰ C, sealed tube.

General Scheme V:

X = Br, F, Cl

Conditions: a) NBS or NCS, MeCN or Selectfluor™, MeCN, water, 0 °C-rt; b) MnO ₂ , CHCl ₃ , heat.

General Scheme VI:

Conditions: a) Pd(OAc) ₂ , PPh ₃ , Cs ₂ CO ₃ , l,4-dioxane:EtOH (2: 1), 140 ⁰ C; b) 2,4-thiazolidinedione, DIEA, AcOH, MW, 170 ⁰ C.

General Scheme VII:

Conditions: a) 2,4-thiazolidinedione, NaOAc, i-PrOH, 170 ⁰ C, MW; b) NHR'R", paraformaldehyde, DIEA, DMF, 95 ⁰ C; then A, Sc(SO ₃ CF ₃ ) ₃ , 95 ⁰ C.

EXAMPLES:

Example 1 :

Preparation of ethyl 6-|YZ)-(2,4-dioxo-l,3-thiazoridin-5- ylidene)methyl]imidazo[ 1 ,2-α ^" |pyridine-3-carboxylate a) Ethyl 6-bromoimidazo[l,2-α]pyridine-3-carboxylate

To a solution of cone. H ₂ SO ₄ (2.7 mL) in EtOH (140 mL) were added 2- amino-5-bromopyridine (Aldrich, 5.92 g, 34.2 mmol) and ethyl 2-chloro-3- oxopropanoate potassium salt (20.0 g, 106.0 mmol; see Tetrahedron, 56, 7915-7921

(2000)) in a sealed tube. The reaction vessel was purged with N ₂ , sealed and heated to 90 ⁰ C. After 18 h, the resultant slurry was cooled to rt, filtered and concentrated under reduced pressure. The residue was diluted with EtOAc, washed with IN NaOH followed by brine and the combined aqueous layers were back-extracted with 95:5 EtOAc/MeOH. The combined extracts were dried over Na ₂ SO ₄ and decolorizing carbon, filtered through a pad of Celite ^® and concentrated under reduced pressure. The residue was recrystallized from 1 : 1 EtOAc/hexanes (100 mL) to give 5.09 g (55%) of the title compound as an off- white solid. MS(ES)+ m/e 271.2 [M+H] ⁺ .

b) Ethyl 6-ethenylimidazo[l,2-α]pyridine-3-carboxylate

Ethyl 6-bromoimidazo[l,2-α]pyridine-3-carboxylate (1.50 g, 5.58 mmol), potassium vinyltrifiuoroborate (1.49 g, 11.2 mmol), PdCl ₂ (dppf)-CH ₂ Cl ₂ (228 mg, 0.28 mmol), triethylamine (1.60 mL, 11.2 mmol) and n-PrOH (50 mL) were combined in a sealed tube, purged with N ₂ and heated to 100 ⁰ C. After 18 h, the reaction mixture was cooled to rt and concentrated under reduced pressure. The residue was diluted with EtOAc, washed with water and brine and the aqueous layers were back-extracted with EtOAc. The combine extracts were dried over

Na ₂ SO ₄ and decolorizing carbon, filtered through a pad of Celite ^® and concentrated under reduced pressure. The residue was purified by silica gel chromatography (EtOAc) to give 1.11 g (92%) of the title compound as a red solid. MS(ES)+ m/e 217.0 [M+H] ⁺ .

c) Ethyl 6-formylimidazo[l,2-α]pyridine-3-carboxylate

Ethyl 6-ethenylimidazo[l,2-α]pyridine-3-carboxylate (500 mg, 2.31 mmol), OsO ₄ (1.2 mL of a 2.5 wt % solution in t-BuOU, 0.116 mmol), NaIO ₄ (1.98 g, 9.24 mmol), 2,6-lutidine (0.53 mL, 4.62 mmol) and 3:1 dioxane/water (20 mL) were combined and stirred at rt. After 2 h, the resultant slurry was diluted with CHCI ₃ , filtered and the filtrate was washed with water. The aqueous layer was extracted with CHCI ₃ and the combined extracts were dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The residue was purified by silica gel chromatography (97:3 CHCl ₃ /MeOH) to give 417 mg (82%) of the title compound as a black solid. MS(ES)+ m/e 219.0 [M+H] ⁺ .

d) Ethyl 6-[(Z)-(2,4-dioxo-l,3-thiazolidin-5-ylidene)methyl]imidazo[l ,2-α]pyridine- 3-carboxylate

Ethyl 6-formylimidazo[l,2-α]pyridine-3-carboxylate (200 mg, 0.917 mmol), 2,4- thiazolidinedione (107 mg, 0.917 mmol), piperidine (91 uL, 0.917 mmol), glacial AcOH (52 uL, 0.917 mmol) and EtOH (5 mL) were combined in a sealed tube, purged with N ₂ and heated to 90 ⁰ C. After 5 h, the reaction was cooled to rt and the product was collected by filtration, giving 82 mg (28%) of the title compound as a tan solid. MS(ES)+ m/e 317.9 [M+H] ⁺ .

Preparation of 6-IYZ)-(2,4-dioxo-l ,3-thiazolidin-5-ylidene)methvH--/V- ethylimidazol ^" 1 ,2-αlpyridine-3-carboxamide

a) 6-Bromo-λ/-ethylimidazo[l ,2-α]pyridine-3-carboxamide

A suspension of ethyl 6-bromoimidazo[l,2-α]pyridine-3-carboxylate (described above, 590 mg, 2.19 mmol) in EtNH ₂ (70% solution in water) was heated to 50 ⁰ C in a sealed tube. After 20 h, the reaction mixture (a clear yellow solution) was cooled to rt and water was added. The resultant white precipitate was collected by filtration and dried under vacuum, providing 475 mg (80%) of the title compound which required no further purification. MS(ES)+ m/e 269.9 [M+H] ⁺ .

b) 6-Ethenyl-JV-ethylimidazo[l ,2-α]pyridine-3-carboxamide

6-Bromo-λ/-ethylimidazo[l,2-α]pyridine-3-carboxamide (402 mg, 1.50 mmol), potassium vinyltrifluoroborate (402 mg, 3.0 mmol), PdCl2(dppf)-CH ₂ Cl2 (61 mg, 0.08 mmol), triethylamine (0.42 mL (3.0 mmol) and n-PrOH (15 mL) were combined in a sealed tube, purged with N ₂ and heated to 100 ⁰ C. After 4 h, the reaction mixture was cooled to rt and concentrated under reduced pressure. The residue was purified by silica gel chromatography (96.5:3.0:0.5

CH ₂ Cl ₂ /MeOH/conc. NH ₄ OH) to give 287 mg (89%) of the title compound as a tan foam. MS(ES)+ m/e 216.0 [M+H] ⁺ .

c) λ/-Ethyl-6-formylimidazo[ 1 ,2-α]pyridine-3-carboxamide

6-Ethenyl-λ/-ethylimidazo[l,2-α]pyridine-3-carboxamide (287 mg, 1.33 mmol), OsO ₄ (0.68 mL of a 2.5 wt % solution in t-BuOU, 0.116 mmol), NaIO ₄ (1.14 g, 9.24 mmol), 2,6-lutidine (0.31 mL, 4.62 mmol) and 3:1 dioxane/water (12 mL) were combined and stirred at rt. After 72 h, the resultant slurry was diluted with CHCI ₃ filtered and the filtrate was washed with water. The aqueous layer was extracted with CHCI ₃ (3X) followed by EtOAc (3X) and the combined extracts were dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The residue was purified by silica gel chromatography (1.5 to 3.0% MeOH in CHCI ₃ ) to give 220 mg (76%) of the title compound as a tan solid. MS(ES)+ m/e 218.0 [M+H] ⁺ .

d) 6-[(Z)-(2,4-Dioxo-l,3-thiazolidin-5-ylidene)methyl]-N-ethyli midazo[l,2- α]pyridine-3-carboxamide

λ/-Ethyl-6-formylimidazo[l,2-α]pyridine-3-carboxamide (160 mg, 0.0.74 mmol), 2,4-thiazolidinedione (130 mg, 1.11 mmol), piperidine (110 uL, 1.11 mmol), glacial AcOH (64 uL, 1.11 mmol) and EtOH (4 mL) were combined in a sealed tube, purged with N ₂ and heated to 90 ⁰ C. After 18 h, the reaction was cooled to rt and the product was collected by filtration. The residue was purified by reverse phase HPLC (MeCN/water) to give 15 mg (6%) of the title compound as an off- white solid. MS(ES)+ m/e 316.8 [M+H] ⁺ .

Preparation of (5 Z)-5-{ [3 -(4-pyridinyl)imidazo[l,2-αlpyridin-6-yllmethylidene I- l,3-thiazoridine-2,4-dione sodium salt

a) 6-Bromoimidazo[l,2-α]pyridine

2-Amino-5-bromopyridine (Aldrich, 18.8 g, 108.7 mmol), chloroacetaldehyde (34 mL of 50% solution in water, 217.4 mmol), NaHCO ₃ (20.1 g, 239.1 mmol) and i-PrOH (150 mL) were combined in a sealed tube, purged with N ₂ , sealed and heated to 80 ⁰ C. After 72 h, the mixture was cooled to rt and concentrated under reduced pressure. The residue was diluted with EtOAc, washed with water and brine and the combined aqueous layers were back-extracted with EtOAc. The combined extracts were dried over Na ₂ SO ₄ and decolorizing carbon, filtered through a pad of Celite ^® and concentrated under reduced pressure. The residue was purified by silica gel chromatography (75 to 85% EtOAc/hexanes to give 17.85 g (83%) of the title compound as a tan solid. MS(ES)+ m/e 198.7 [M+H] ⁺ .

b) 6-Bromo-3-iodoimidazo[l ,2-α]pyridine

To a slightly cloudy mixture of 6-bromoimidazo[l,2-α]pyridine (15.4 g, 78.0 mmol) in dry MeCN (500 mL) was added jV-iodosuccinimide (17.6 g, 78.0 mmol) portionwise over ~5 min (a precipitate immediately formed). After 16 h, the crude product was collected by filtration and triturated with boiling MeCN to provide 19.56 g (77%) of the title compound as an off-white solid. MS(ES)+ m/e 198.7 [M+H] ⁺ .

c) 6-Bromo-3-(4-pyridinyl)imidazo[l ,2-α]pyridine

6-Bromo-3-iodoimidazo[l,2-α]pyridine (6.57 g, 20.3 mmol), pyridine 4- boronic acid (2.50 g, 20.3 mmol), Pd(PPh ₃ ) ₄ (1.17 g, 1.01 mmol) aq. K ₂ CO ₃ (8.42 g, 61.0 mmol in 50 mL of water) and 1,4-dioxane (150 mL) were combined in a sealed tube and heated to 100 ⁰ C. After 16 h, the reaction was cooled to rt and concentrated under reduced pressure. The solid residue was triturated with water, collected by filtration and azeotroped with CH ₂ Cl ₂ . The crude product dissolved in boiling CHCl ₃ , and insoluble impurities were removed by filtration. Concentration of the filtrate followed by silica gel chromatography (0 to 3 to 5% MeOH/CHCl ₃ ) provided 2.12 g (38%) of the title compound as a yellow solid. MS(ES)+ m/e 275.9 [M+H] ⁺ .

d) 6-Ethenyl-3-(4-pyridinyl)imidazo[ 1 ,2-α]pyridine

6-Bromo-3-(4-pyridinyl)imidazo[l,2-α]pyridine (1.50 g, 5.47 mmol), potassium vinyltrifluoroborate (1.46 g, 10.9 mmol), PdCl ₂ (dppf)-CH ₂ Cl ₂ (223 mg, 0.274 mmol), triethylamine (1.5 mL, 10.9 mmol) and n-PrOH (50 mL) were combined in a sealed tube, purged with N ₂ and heated to 100 ⁰ C. After 18 h, the reaction mixture was cooled to rt and concentrated under reduced pressure. The residue was diluted with EtOAc, washed with water and brine and the aqueous layers were back-extracted with EtOAc. The combine extracts were dried over Na ₂ SO ₄ and decolorizing carbon, filtered through a pad of Celite ^® and concentrated under reduced pressure. The residue was purified by silica gel chromatography (96.5:3.0:0.5 CH ₂ Cl ₂ /MeOH/conc. NH ₄ OH) to give 1.04 g (85%) of the title compound as a grey solid. MS(ES)+ m/e 221.9 [M+H] ⁺ .

e) 3-(4-Pyridinyl)imidazo[l ,2-α]pyridine-6-carbaldehyde

6-Ethenyl-3-(4-pyridinyl)imidazo[l,2-α]pyridine (476 mg, 1.74 mmol), OsO ₄ (0.88 mL of a 2.5 wt % solution in t-BuOU, 0.0.09 mmol), NaIO ₄ (1.49 g, 6.96 mmol), 2,6-lutidine (0.40 mL, 3.48 mmol) and 3:1 dioxane/water (16 mL) were combined and stirred at 70 ⁰ C in a sealed tube. After 5 h, the resultant slurry was cooled to rt, diluted with CHCI ₃ , filtered and the filtrate was washed with water and brine. The aqueous layer was extracted with CHCI ₃ and the combined extracts were dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The residue was purified by silica gel chromatography (97:3 CHCl ₃ /MeOH) to give 255 mg (53%) of the title compound as an off-white solid. MS(ES)+ m/e 223.8 [M+H] ⁺ .

f) (5Z)-5-{[3-(4-Pyridinyl)imidazo[l,2-α]pyridin-6-yl]methylid ene}-l,3- thiazolidine-2,4-dione sodium salt

3-(4-Pyridinyl)imidazo[l,2-α]pyridine-6-carbaldehyde (200 mg, 0.897 mmol), 2,4- thiazolidinedione (158 mg, 1.35 mmol), piperidine (134 uL, 1.35 mmol), glacial AcOH (77 uL, 1.35 mmol) and EtOH (4 mL) were combined in a sealed tube, purged with N ₂ and heated to 90 ⁰ C. After 16 h, the reaction was cooled to rt and the crude precipitate was collected by filtration and recrystallized from DMSO. A portion of the pure free acid (40 mg, 0.124 mmol) was suspended in THF (4 mL) and treated with 6N NaOH (21 uL, 0.124 mmol). After stirring overnight, the precipitate was collected by filtration to give 35 mg of the title compound as an orange solid. MS(ES)+ m/e 323.0 [M+H] ⁺ .

Example 4

Preparation of (5Z)-5-(Imidazo[ 1 ,2-α]pyridin-6-ylmethylidene)- 1 ,3-thiazolidine-2,4- dione

The title compound was prepared as described for example 1, substituting 6- bromoimidazo[l,2-α]pyridine for ethyl 6-bromoimidazo[l,2-α]pyridine-3- carboxylate, to give the title compound as an off- white solid. MS(ES)+ m/e 245.8 [M+H] ⁺ .

Preparation of (5 Z)-5- [(3 -acetylimidazori^-αlpyridin-ό-vDmethylidenel- 1,3- thiazolidine-2,4-dione

a) 1 -(6-Bromoimidazo[ 1 ,2-α]pyridin-3-yl)ethanone

A mixture of 6-bromo-3-iodoimidazo[l,2-α]pyridine (3.0 g, 9.29 mmol), 1- ethoxyvinyltri-n-butyltin (3.3 mL, 9.75 mmol) and dichlorobis(triphenylphosphine)- palladium(II) (325 mg, 0.46 mmol) in 1,4-dioxane (90 mL) was heated to 100 ⁰ C in a sealed tube for 16 h. The reaction mixture was cooled to rt, diluted with ethyl acetate, filtered through a pad of silica gel (elute with ethyl acetate) and concentrated under reduced pressure. The residue was taken up in THF (50 mL) and treated with 6N HCl (3.1 mL, 18.6 mmol). After 1 h, the mixture was concentrated under reduced pressure, triturated with water and adjusted to pH ~12 with IN NaOH. The mixture was extracted 3 X with ethyl acetate and the combined extracts were dried (Na ₂ SO ₄ ), filtered and concentrated under reduced pressure. The residue was purified by silica gel chromatography (97:3 EtOAc/MeOH) to give 1.41 g (64%) of l-(6-bromoimidazo[l,2-α]pyridin-3-yl)ethanone as a pale yellow solid. MS(ES)+ m/e 240.7 [M+H] ⁺ .

b) (5Z)-5-(Imidazo[ 1 ,2-α]pyridin-6-ylmethylidene)- 1 ,3-thiazolidine-2,4-dione.

The title compound was prepared as described for example X, substituting l-(6- bromoimidazo[l,2-α]pyridin-3-yl)ethanone for ethyl 6-bromoimidazo[l,2- α]pyridine-3-carboxylate, to give the title compound as light yellow solid after recrystallization from glacial AcOH. MS(ES)+ m/e 287.8 [M+H] ⁺ .

Example 6

Preparation of (5 Z)-5-(imidazo[l,2-αlpyrimidin-6-ylmethylidene)-l,3-thiazoli dine- 2,4-dione

a) 6-Bromoimidazo[ 1 ,2-α]pyrimidine

A suspension of 2-amino-5-bromopyrimidine (3.0 g, 17.2 mmol), bromoacetaldehyde diethyl acetal (3.2 mL, 20.7 mmol) and 48% HBr (1.7 mL) in EtOH was heated was heated to 80 ⁰ C in a sealed tube for 16 h. After cooling to rt, the reaction was adjusted to pH ~12 with 6N NaOH and the resultant precipitate was collected by filtration, rinsed with water followed by hexanes and dried to a constant weight to give 2.12 g (62%) of 6-bromoimidazo[l,2-α]pyrimidine as a white solid. MS(ES)+ m/e 199.7 [M+H] ⁺ .

b) (5Z)-5-(Imidazo[ 1 ,2-α]pyrimidin-6-ylmethylidene)- 1 ,3-thiazolidine-2,4-dione.

The title compound was prepared as described for example X, substituting 6- bromoimidazo[l,2-α]pyrimidine for ethyl 6-bromoimidazo[l,2-α]pyridine-3- carboxylate, to give the title compound as tan solid. MS(ES)+ m/e 246.8 [M+H] ⁺ .

Preparation of (5Z)-5-[(3-acetyl-2-methylimidazo[ 1 ,2-α ^" |pyridin-6-yl)methylidenel- 1 ,3-thiazoridine-2,4-dione

To a 25-mL microwave vial were added 2-amino-5-bromopyridine (2.0 g, 11.6 mmol), 3-chloroacetylacetone (1.4 mL, 12.7 mmol) and n-propanol (12 mL). The vial was purged with nitrogen, sealed and heated in a microwave reactor at 200 ⁰ C for 30 min. The reaction mixture was filtered and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel chromatography (EtOAc) followed by trituration with boiling 1 : 1 EtOAc/hexanes. Upon cooling to room temperature, the precipitate was collected to give 620 mg (21%) of l-(6- bromo-2-methylimidazo[l,2-α]pyridin-3-yl)ethanone as a tan solid. MS(ES)+ m/e 255 [M+H] ⁺ . l-(6-Bromo-2-methylimidazo[l,2-α]pyridin-3-yl)ethanone was converted to the title compound utilizing procedures detailed for example 1, thereby providing (5Z)-5- [(3 -acetyl-2-methylimidazo[l,2-α]pyridin-6-yl)methylidene]- 1,3- thiazolidine-2,4-dione as a yellow solid. MS(ES)+ m/e 302 [M+H] ⁺ .

Preparation of (5 Z)S- { [3 -(trifluoromethyl)imidazo [1, 2-α]pyridin-6- yl]methyridene|-l,3-thiazoridine-2,4-dione

Example 8 was prepared as described in example 3, substituting 3-bromo-l,l,l- trifluoro-2-propanone for chloroacetaldehyde in step (a), to give the title compound as an off-white solid. MS(ES)+ m/e 313.9 [M+H] ⁺ .

Preparation of 6-[(Z)-(2,4-dioxo-l,3-thiazolidin-5-ylidene)methyllimidazo[ 1,2- αlpyridine-3-carbonitrile acetate

a) 7V-(5 -bromo-2-pyridinyl)-N,λ/-dimethylimidoformamide

To a solution of 2-amino-5-bromopyridine (5.0 g, 28.9 mmol), in dry MeOH was added DMF-DMA (12.7 mL, 95.4 mmol) in a sealable reaction tube. The

reaction was purged with nitrogen, sealed and heated to 70 ⁰ C. After 5.5 h, the reaction mixture was cooled to room temperature and concentrated under reduced pressure. The resulting solid was recrystallized from hexanes to give 4.1 g (62%) of ^-(S-bromo-l-pyridiny^-^N-dimethylimidoformamide as a yellow solid. MS(ES)+ m/e 228 [M+H] ⁺ .

b) 6-bromoimidazo[ 1 ,2-α]pyridine-3-carbonitrile

To a mixture of λ/'-(5-bromo-2-pyridinyl)-N,λ/-dimethylimidoformamide (3.8 g, 16.7 mmol) in i-PrOH (80 mL) was added bromoacetonitrile (2.3 mL, 33.4 mmol) followed by NaHCO ₃ (3.5 g, 41.8 mmol) in a sealable reaction tube. The reaction was purged with nitrogen, sealed and heated to 100 ⁰ C. After 1 h, the reaction mixture was concentrated under reduced pressure and the residue was suspended in water (100 mL). The precipitate was collected by filtration and triturated with boiling acetonitrile to give 1.39 g (37%) of 6-bromoimidazo[l,2-α]pyridine-3- carbonitrile as a brown solid. MS(ES)+ m/e 222.8 [M+H] ⁺ .

Example 9 was prepared as described in example 1 , substituting 6- bromoimidazo[l,2-α]pyridine-3-carbonitrile in place of ethyl 6-bromoimidazo[l,2- α]pyridine-3-carboxylate in step (b), to give the title compound as an acetic acid solvate (off-white solid). MS(ES)+ m/e 270.9 [M+H] ⁺ .

Example 10:

Preparation of (5Z)-5- {[3-(2,2-dimethylpropanoyl)imidazo[l ,2-α]pyridin-6- yl]methyridene|-l,3-thiazoridine-2,4-dione

Example 10 was prepared as described for example 9, substituting l-bromo-3,3- dimethyl-2-butanone in place of bromoacetonitrile in step (b) and following the remainder of the reaction sequence detailed for example 1, to give the title compound as a pale yellow solid. MS(ES)+ m/e 330.1 [M+H] ⁺ .

Example 11 :

Preparation of (5Z)-5-[(3-propanoyrimidazo| ^" 1 ,2-α1pyridin-6-yl)methylidene]- 1 ,3- thiazolidine-2,4-dione

Example 11 was prepared as described for example 9, substituting l-bromo-2- butanone in place of bromoacetonitrile in step (b) and following the remainder of the reaction sequence detailed for example 1 , to give the title compound as a tan solid. MS(ES)+ m/e 302.1 [M+H] ⁺ .

Example 12:

Preparation of 2- {6-r(Z)-(2,4-dioxo-l,3-thiazolidin-5-ylidene)methyllimidazor i, 2- αlpyridin-3-vU -2-oxoethyl acetate

Example 12 was prepared as described for example 9, substituting 3-chloro-2- oxopropyl acetate in place of bromoacetonitrile in step (b) and following the remainder of the reaction sequence detailed for example 1, to give the title compound as an off-white solid. MS(ES)+ m/e 345.9 [M+H] ⁺ .

Example 13:

Preparation of (5 Z)-5-{ [3 -(I -hvdroxypropyl)imidazori,2-αlpyridin-6-

yl1methyridene|-l,3-thiazoridine-2,4-dione

(5Z)-5- {[3 -(I -Hydroxypropyl)imidazo[ 1 ,2-α]pyridin-6-yl]methylidene} - 1 ,3- thiazolidine-2,4-dione (example 13) was prepared through reduction of (5Z)-5-[(3- propanoylimidazo[l,2-α]pyridin-6-yl)methylidene]-l,3-thiazo lidine-2,4-dione using the following procedure: To a suspension of (5Z)-5-[(3-propanoylimidazo[l,2- α]pyridin-6-yl)methylidene]-l,3-thiazolidine-2,4-dione (200 mg, 0.66 mmol) in MeOH (14 niL) was added IN NaOH (0.66 mL, 0.66 mmol), followed by NaBH ₄ (12.6 mg, 0.33 mmol). After 18 h, an additional portion OfNaBH ₄ (6.3 mg, 0.17 mmol) was added. After 2 h, the reaction mixture was concentrated under reduced pressure and the residue was triturated with water and adjusted to pH 7 with IN HCl. Concentration of the filtrate provided a yellow solid, which was taken up in water, acidified to pH 6.5 with IN HCl and allowed to cool to room temperature. The resulting precipitate was collected to give 14 mg (7%) of the title compound as a yellow solid. MS(ES)+ m/e 303.9 [M+H] ⁺ .

Preparation of (5 Z)-5-{ [3 -(hvdroxyacetyl)imidazori,2-alpyridin-6-yllmethylidene I - 1 ,3-thiazoridine-2,4-dione

(5Z)-5-([3-(Hydroxyacetyl)imidazo[l,2-α]pyridin-6-yl]methyl idene}-l,3- thiazolidine-2,4-dione (example 14) was prepared through hydrolysis of 2-(6-[(Z)- (2,4-dioxo- 1 ,3-thiazolidin-5-ylidene)methyl]imidazo[ 1 ,2-α]pyridin-3-yl} -2-oxoethyl acetate using the following procedure: To a suspension of 2-{6-[(Z)-(2,4-dioxo-l,3- thiazolidin-5-ylidene)methyl]imidazo[ 1 ,2-α]pyridin-3-yl} -2-oxoethyl acetate (60 mg, 0.17 mmol) in MeOH (2 mL) was added IN NaOH (0.35 mL, 0.35 mmol). The reaction mixture briefly became clear, then a fine suspension formed. After 1 h, the reaction mixture was allowed to cool to 0 ⁰ C and the precipitate was collected and rinsed with a minimal amount of cold MeOH to provide 48 mg (84%) of the title compound as a sodium salt. MS(ES)+ m/e 303.9 [M+H] ⁺ .

Example 15:

Preparation of (5 Z)-5-{[3-(5-methyl-l,2,4-oxadiazol-3-yl)imidazo[l,2-αlpyrid in-6- yl]methyridene|-l,3-thiazoridine-2,4-dione

a) 6-ethenylimidazo[l ,2-α]pyridine-3-carbonitrile

A sealable reaction tube was charged with 6-bromoimidazo[l,2-α]pyridine-

3-carbonitrile (1.39 g, 6.26 mmol), tri-n-butylvinylstannane (1.9 mL, 6.57 mmol), Pd(PPhs)4 (362 mg, 0.31 mmol) and 1,4-dioxane (60 mL). The reaction was purged with nitrogen, sealed and heated to 100 ⁰ C. After 3.5 h, the reaction was allowed to cool to room temperature, filtered and concentrated under reduced pressure. The residue was purified by silica gel chromatography (35% EtOAc in hexanes) to give 903 mg (85%) of 6-ethenylimidazo[l,2-α]pyridine-3-carbonitrile as a white solid. MS(ES)+ m/e 169.8 [M+H] ⁺ .

b) 6-ethenyl-JV-hydroxyimidazo[l ,2-α]pyridine-3-carboximidamide

To a suspension of 6-ethenylimidazo[l,2-α]pyridine-3-carbonitrile (340 mg, 2.01 mmol) in EtOH (20 mL), was added triethylamine (0.42 mL, 3.02 mmol) followed by hydroxylamine hydrochloride (153 mg, 2.21 mmol). After several minutes, the suspension became a solution. After 18 h, the reaction mixture was concentrated to dryness and the resulting solid was suspended in water, stirred vigorously and the precipitate was collected to give 324 mg (80%) of 6-ethenyl-JV- hydroxyimidazo[l,2-α]pyridine-3-carboximidamide as an off-white solid. MS(ES)+ m/e 203.1 [M+H] ⁺ .

c) 6-ethenyl-3-(5-methyl-l,2,4-oxadiazol-3-yl)imidazo[l,2-α]py ridine

To a suspension of 6-ethenyl-λ/-hydroxyimidazo[l,2-α]pyridine-3- carboximidamide (295 mg, 1.46 mmol) in toluene (15 mL) was added acetic

anhydride (1.4 mL, 14.6 mmol) in a sealbale reaction tube. The reaction mixture was purged with nitrogen, sealed and heated to 120 ⁰ C. After 7 h, the reaction mixture was allowed to cool to room temperature, filtered and concentrated under reduced pressure. The resulting residue was purified by silica gel chromatography (EtOAc) to provide 238 mg (72%) of 6-ethenyl-3-(5-methyl-l,2,4-oxadiazol-3- yl)imidazo[l,2-α]pyridine as an off-white solid. MS(ES)+ m/e 226.9 [M+H] ⁺ .

Example 15 was prepared as described for example 1, substituting 6-ethenyl-3-(5- methyl-l,2,4-oxadiazol-3-yl)imidazo[l,2-α]pyridine in place of ethyl 6- formylimidazo[l,2-α]pyridine-3-carboxylate in step (c) and following the remainder of the reaction sequence detailed for example 1, to give the title compound as whitesolid. MS(ES)+ m/e 328.0 [M+H] ⁺ .

Preparation of (5Z)-5-r(3-bromoimidazor 1 ,2-αlpyridin-6-yl)methylidenel- 1,3- thiazolidine-2,4-dione acetate

a) (3-bromoimidazo[ 1 ,2-α]pyridin-6-yl)methanol

To a suspension of imidazo[l,2-α]pyridin-6-ylmethanol (1.0 g, 6.75 mmol,

Maybridge) in acetonitrile (30 mL) was added NBS (1.20 g, 6.75 mmol) in one portion. The mixture became clear, then a precipitate formed. After 1 h, the reaction mixture was concentrated under reduced pressure and the residue was triturated with acetonitrile (20 mL). The precipitate was collected by filtration to give 1.03 g (67%) of (3-bromoimidazo[l,2-α]pyridin-6-yl)methanol as a red solid. MS(ES)+ m/e 226.8 [M+H] ⁺ .

b) 3-bromoimidazo[ 1 ,2-α]pyridine-6-carbaldehyde

To a suspension of (3-bromoimidazo[l,2-α]pyridin-6-yl)methanol (300 mg, 1.32 mmol) in CHCl ₃ (25 mL) was added MnO ₂ (1.15 g, 13.2 mmol). The reaction was heated to 70 ⁰ C for 2 h, filtered through a pad of Celite ^® and concentrated under reduced pressure to give 242 mg (81%) of 3-bromoimidazo[l,2-α]pyridine-6-

carbaldehyde as a white solid. MS(ES)+ m/e 224.8 [M+H] ⁺ .

Example 16 was prepared as described for example 1, substituting 3- bromoimidazo[l,2-α]pyridine-6-carbaldehyde in place of ethyl 6- formylimidazo[l,2-α]pyridine-3-carboxylate in step (d) to give the title compound as an off-white solid. MS(ES)+ m/e 323.9 [M+H] ⁺ .

Preparation of (5Z)-5-[(3-fluoroimidazo[ 1 ,2-α1pyridin-6-yl)methylidene ^" |- 1,3- thiazolidine-2,4-dione

a) (3-fluoroimidazo[ 1 ,2-α]pyridin-6-yl)methanol

To a cold (0 ⁰ C) slurry of imidazo[l,2-α]pyridin-6-ylmethanol (300 mg, 2.02 mmol) in acetonitrile (20 mL) was added a solution of Selectfluor™ (358 mg, 1.01 mmol) in 1 : 1 v/v THF/water (7 mL) dropwise over 30 min. The resulting solution was allowed to warm to rt, stirred for 2 h, and concentrated under reduced pressure. The residue was purified by silica gel chromatography (95:5 EtOAc/MeOH) to give 92 mg (27%) of (3-fluoroimidazo[l,2-α]pyridin-6-yl)methanol as an off- white solid. MS(ES)+ m/e 166.8 [M+H] ⁺ .

Example 17 was prepared as described for example 16, susbstituting (3- fluoroimidazo[ 1 ,2-α]pyridin-6-yl)methanol in place of (3-bromoimidazo[ 1 ,2- α]pyridin-6-yl)methanol in step (b), then following the procedure described for example 1, substituting 3-fluoroimidazo[l,2-α]pyridine-6-carbaldehyde in place of ethyl 6-formylimidazo[l,2-α]pyridine-3-carboxylate in step (d) to give the title compound as a dark tan solid. MS(ES)+ m/e 263.8 [M+H] ⁺ .

Example 18:

Preparation of (5Z)-5-[(3-chloroimidazo[ 1 ,2-αlpyridin-6-yl)methylidene ^" |- 1 ,3- thiazolidine-2,4-dione

a) (3-chloroimidazo[l,2-α]pyridin-6-yl)methanol

To a suspension of imidazo[l,2-α]pyridin-6-ylmethanol (410 mg, 2.77 mmol, Maybridge) in acetonitrile (14 mL) was added NCS (388 mg, 2.91 mmol) in one portion. After 4 h, the reaction mixture was concentrated under reduced pressure and the residue was triturated with boiling 95:5 v/v EtOAc/MeOH and hot filtered. The filtrate was purified directly by silica gel chromatography (95:5 EtOAc/MeOH) to give 343 mg (68%) of (3-chloroimidazo[l,2-α]pyridin-6-yl)methanol as a pale yellow solid. MS(ES)+ m/e 182.7 [M+H] ⁺ .

b) 3-chloroimidazo[l ,2-α]pyridine-6-carbaldehyde

To a suspension of (3-chloroimidazo[l,2-α]pyridin-6-yl)methanol (340 mg, 1.86 mmol) in CHCl ₃ (10 mL) was added MnO ₂ (1.62 g, 18.6 mmol). The reaction was heated to 50 ⁰ C for 2.5 h, filtered through a pad of Celite ^® and concentrated under reduced pressure to give 272 mg (80%) of 3-chloroimidazo[l,2-α]pyridine-6- carbaldehyde as a white solid.

Example 18 was prepared as described for example 16, substituting 3- chloroimidazo[l,2-α]pyridine-6-carbaldehyde in place of ethyl 6- formylimidazo[l,2-α]pyridine-3-carboxylate in step (d) to give the title compound as a yellow solid. MS(ES)+ m/e 279.8 [M+H] ⁺ .

Example 19:

Preparation of (5Z)-5-{r3-(4-isoquinolinyl)imidazori,2-α1pyridin-6- yl1methyridene|-l,3-thiazoridine-2,4-dione

a) 3-(4-isoquinolinyl)imidazo[ 1 ,2-α]pyridine-6-carbaldehyde

A suspension of imidazo[l,2-α]pyridine-6-carbaldehyde (Maybridge, 0.1 g, 0.68 mmol), 4-bromoisoquinoline (Oakwood, 0.214 g, 1.03 mmol), triphenylphosphine (0.013g, 0.05 mmol), Pd(OAc) ₂ (0.006g, 0.03 mmol), Cs ₂ CO ₃ (0.454g, 1.39 mmol), and 1,4-dioxane: EtOH (2:1) (2 mL) was capped and heated to 140 ⁰ C in a sealed vial for 2 days. Additional Dioxane (1 mL), DMF (1 mL), 4-bromoisoquinoline (Oakwood, 0.214 g, 1.03 mmol), triphenylphosphine (0.013g, 0.05 mmol), and Pd(OAc) ₂ (0.006g, 0.03 mmol) were added and the mixture was again heated between 90-110 ⁰ C until judged not proceed any further. Cooled the reaction to rt and diluted with EtOAc, filtered through a pad of celite eluding with DMF, then concentrated in vacuo. The crude mixture was taken on without further purification

b) (5Z)-5-{[3-(4-isoquinolinyl)imidazo[l,2-α]pyridin-6-yl]meth ylidene}-l,3- thiazolidine-2,4-dione.

The crude 3-(4-isoquinolinyl)imidazo[l,2-α]pyridine-6-carbaldehyde, 2,4- thiazolidinedione (83 mg, 0.71 mmol), sodium acetate ( 174 mg, 2.12 mmol), and glacial AcOH ( 2 mL) were combined in a 2-5 mL Biotage microwave vial, then sealed and heated in a Biotage Initiator microwave system (0-400 W) to 170 ⁰ C for Ih. Filtered the resulting suspension after cooling to rt washing the solid with glacial AcOH and MeOH before HPLC purification afforded (5Z)-5- {[3-(4- isoquinolinyl)imidazo[l,2-α]pyridin-6-yl]methylidene}-l,3-t hiazolidine-2,4-dione as a brown solid (0.058 g, 22%). MS(ES)+ m/e 373 [M+H] ⁺ .

Example 20:

Preparation of (5 Z)-5-{ [3 -(4-isoquino linyl)imidazo[ l,2-αlpyridin-6- vHmethyridene|-l,3-thiazoridine-2,4-dione

The title compound was prepared as described for example 19, substituting 2- bromopyridine for 4-bromoisoquinoline, to give the title compound as a yellow solid (0.01 g, 4%). MS(ES)+ m/e 323 [M+H] ⁺ .

Example 21 :

Preparation of (5Z)-5-{r3-(3-pyridinyl)imidazori,2-αlpyridin-6-yllmethylid ene|- 1 ,3-thiazoridine-2,4-dione

The title compound was prepared as described for example 19, substituting 3- bromopyridine for 4-bromoisoquinoline, to give the title compound as an orange solid (0.025 g, 11%). MS(ES)+ m/e 323 [M+H] ⁺ .

Preparation of ethyl 5-{6-r(Z)-(2,4-dioxo-l,3-thiazolidin-5- ylidene)methyllimidazor 1 ,2-αlpyridin-3-yl| -3-pyridinecarboxylate

The title compound was prepared as described for example 19, substituting methyl 5-bromonicotinate for 4-bromoisoquinoline, to give, after transesterifϊcation with the EtOH in the first step, the title compound as a yellow solid (0.012 g, 4%). MS(ES)+ m/e 395 [M+H] ⁺ .

Example 23:

Preparation of (5 Z)-5-{ [3 -(6-methyl-2-pyridinyl)imidazoπ,2-αlpyridin-6- yl1methyridene|-l,3-thiazoridine-2,4-dione

The title compound was prepared as described for example 19, substituting 2- bromo-6-methylpyridine for 4-bromoisoquinoline, to give the title compound as a pale yellow solid (0.004 g, 2%). MS(ES)+ m/e 337 [M+H] ⁺ .

Preparation of (5Z)-5-\(3- {\(l , 1 -dioxido- 1 -benzothien-6- yl)aminolmethyUimidazo[l,2-αlpyridin-6-yl)methylidenel-l,3- thiazolidine-2,4- dione

a) (5Z)-5-(Imidazo[l,2-α]pyridin-6-ylmethylidene)-l,3-thiazoli dine-2,4-dione.

A suspension of imidazo[l,2-α]pyridine-6-carbaldehyde (Maybridge, 1.04 g, 7.1 mmol), thiazolidinedione (0.98 g, 8.3 mmol), sodium acetate ( 1.82 g, 22.2 mmol), and glacial AcOH ( 5 mL) were combined in a 20 mL Biotage microwave vial, then sealed and heated in a Biotage Initiator microwave system (0-400 W) to 170 ⁰ C for Ih. After cooling to rt, added an additional 5 mL glacial acetic acid, sonicated the mixture and filtered the resulting suspension. Washed the solid with glacial AcOH and MeOH to afford, after drying the solid in vacuo, (5Z)-5-(Imidazo[l,2-α]pyridin- 6-ylmethylidene)-l,3-thiazolidine-2,4-dione as a yellow solid (1.72 g, 82%). MS(ES)+ m/e 246 [M+H] ⁺ .

b) (5Z)-5-[(3-{[(l,l-dioxido-l-benzothien-6-yl)amino]methyl}imi dazo[l,2- α]pyridin-6-yl)methylidene]- 1 ,3-thiazolidine-2,4-dione.

1,1 -dioxido- l-benzothien-6-yl)amine (Frontier, 0.073 g, 0.4 mmol), paraformaldehyde (0.03g, 1 mmol), diisoproplyethylamine (0.2 mL, 3.73 mmol), and DMF (1 mL) were combined in a 2 dram vial, capped, and heated to 95 ⁰ C

overnight. The reaction was cooled to rt then (5Z)-5-(imidazo[l,2-α]pyridin-6- ylmethylidene)-l,3-thiazolidine-2,4-dione (0.06g, 0.24 mmol) and scandium trifluoromethanesulfonate (0.01 mg 0.02 mmol) were added and the vial was sealed and heated again to 95 ⁰ C overnight. When judged to be complete, the reaction was cooled to rt, water was added (2 mL) and the resulting suspension was filtered and washed with water. The solid purified by RP HPLC to afford (5Z)-5-[(3-{[(l,l- dioxido-l-benzothien-6-yl)amino]methyl}imidazo[l,2-α]pyridi n-6-yl)methylidene]- l,3-thiazolidine-2,4-dione as an orange solid ( 0.013 g, 7%). MS(ES)+ m/e 439 [M+H] ⁺ .

Preparation of (5Z)-5 - { [3 -( { [3 -(methylsulfonvDphenvH amino I methvDimidazo [1,2- α1pyridin-6-yl1methylidene|-l,3-thiazoridine-2,4-dione

The title compound was prepared as similarly to the procedure described for example 24, substituting 3-(methylsulfonyl)aniline (Aldrich) for 1,1-dioxido-l- benzothien-6-yl)amine, to give the title compound as an orange solid (0.010 g, 5%). MS(ES)+ m/e 429 [M+H] ⁺ .

Preparation of (5Z)-5 - |Y3 - { \4-( (3 - [(trifluoromethyl)sulfonyl]phenyU amino)- 1 - piperidinyllmethyl|imidazo[l,2-αlpyridin-6-yl)methylidenel- l,3-thiazolidine-2,4- dione

The title compound was prepared as similarly to the procedure described for

example 24, substituting N-(3-[(trifluoromethyl)sulfonyl]phenyl)-4-piperidinamine for l,l-dioxido-l-benzothien-6-yl)amine, and after RP HPLC purification, dissolving in dichloromethane, washing with 1 N NaHCC> ₃ , and concentrating in vacuo the organics to afford (5Z)-5-[(3-{[4-({3- [(trifluoromethyl)sulfonyl]phenyl} amino)- 1 -piperidinyljmethyl} imidazo[ 1 ,2- α]pyridin-6-yl)methylidene]-l,3-thiazolidine-2,4-dione as an off- white solid (0.01 g, 4%). MS(ES)+ m/e 566 [M+H] ⁺ .

Exemplary capsule composition An oral dosage form for administering the present invention is produced by filing a standard two piece hard gelatin capsule with the ingredients in the proportions shown in Table I, below.

Table I

INGREDIENTS AMOUNTS compound of example 1 25 mg

Lactose 55 mg

Talc 16 mg

Magnesium Stearate 4 mg

Exemplary Injectable Parenteral Composition

An injectable form for administering the present invention is produced by stirring 1.5% by weight of compound of example 1 in 10% by volume propylene glycol in water.

Exemlary Tablet Composition

The sucrose, calcium sulfate dihydrate and an PI3K inhibitor as shown in

Table II below, are mixed and granulated in the proportions shown with a 10% gelatin solution. The wet granules are screened, dried, mixed with the starch, talc and stearic acid;, screened and compressed into a tablet.

Table II

INGREDIENTS AMOUNTS compound of example 1 20 mg calcium sulfate dehydrate 30 mg

Sucrose 4 mg

Starch 2 mg Talc 1 mg stearic acid 0.5 mg

While the preferred embodiments of the invention are illustrated by the above, it is to be understood that the invention is not limited to the precise instructions herein disclosed and that the right to all modifications coming within the scope of the following claims is reserved.