HSP90 MODULATING COMPOUNDS, COMPOSITIONS, METHODS AND USES

Cross-Reference to Related Application

This application claims the benefit of and priority to U.S. Provisional Patent Application No. 61/245,951 filed September 25, 2009 and U.S. Provisional Patent Application No. 61/371 ,482 filed August 6, 2010.

The content of the above patent application is hereby expressly incorporated by reference into the detailed description hereof.

Background of the Invention

The heat shock protein 90 family, including HSP90 alpha, HSP90 beta, HSP90 N,

GRP94 and TRAP- 1 (herein generically referred to as HSP90), are essential molecular chaperones that participate in folding of newly synthesized proteins and stabilization and refolding of stress-denatured client proteins. A large number of HSP90 client proteins have been identified many of which are involved in cellular signaling. Consequently, inhibition of HSP90 is widely regarded as a promising therapeutic approach for the treatment of diseases and conditions characterized by inappropriate cellular signaling and stress responses including cancer, neurodegeneration, inflammation and autoimmune disease.

HSP90 Structure, Sub-cellular Localization and Function

HSP90 and analogues are relatively large proteins that have characteristic

Bergerat ATP binding folds and belong to the GHKL protein superfamily of ATPases and protein kinases (Dutta, R. and Inouye, M., Trends Biochem Sci., 2000, 25( 1 ): 24-8). Crystallography studies revealed a pocket in the N-terminal domain with sequence homology to type II topoisomerases and MutL mismatch repair proteins which co- crystallized with ATP/ADP and established that the N-terminal region is a binding site for adenine nucleotides (Prodromou, C. et al., Cell, 1997, 90( 1 ): 65-75). Additionally, HSP90 has a middle segment that participates in client protein binding and a c-terminal domain that is responsible for dimerization and binding of some natural products including novobiocin (Pearl, L. H. and Prodromou, C, Annu Rev Biochem, 2006, 75: 271 - 94). HSP90 is one of the most abundant cellular proteins comprising 1 -2% of total cellular protein (Iwasaki, M. et al., Biochim Biophys Acta, 1 89, 992( 1 ): 1 -8). HSP90 alpha and HSP90 beta are dimeric cytosolic proteins whereas GRP94 and TRAP- 1 reside in the endoplasmic reticulum and mitochondria respectively. HSP90 binds to a series of co-chaperones in an ATP dependent manner, thereby modulating the structure of client proteins. ATP binding regulates the conformation of HSP90 modulating interactions with client proteins (Richter, K., et al., ./ Biol Chem, 2008, 283(26): 17757-65).

HSP90 exerts its cellular function through selectively chaperoning, or promoting conformational changes and domain rearrangements in a wide range of client proteins including nuclear hormone receptors (Pratt, W. B. and Toft, D. O., Exp Biol Med

(Maywood), 2003, 228(2): 1 1 1 -33) and protein kinases (Pearl, L. H., Curr. Opin. Genet. Dev. , 2005, 15( 1 ): 55-61 ). Pharmacological inhibition of HSP90 results in induction of a heat shock response as well as destabilization of HSP90 client proteins. In contexts where pathology is driven by proteins that are HSP90 clients it is expected that HSP90 inhibition will result in destabilization of key proteins leading to therapeutic benefit. It is also expected that induction of a heat shock response, including enhanced synthesis of heat shock protein 70 (HSP70) (Lu, A. et al., ./. Neurochem., 2002, 81 (2): 355-64), following inhibition of HSP90 may protect normal cells from inappropriate toxicity.

Various natural product small molecules have been identified which bind to the N-terminal ATP binding site of HSP90, including geldanamycin (Stebbins, C. E. et al., Cell, 1997, 89(2): 239-50 and Whitesell, L. et al., Proc. Natl. Acad. Sci. U S A, 1994, 91 ( 18): 8324-8) and related benzoquinone ansamycins.

HSP90 and the immune system

Inflammation and autoimmune diseases are characterized by cellular events that are mediated through signaling proteins many of which are HSP90 clients. The NF- kappa B pathway plays an important role in inflammation and autoimmune disease (Vallabhapurapu, S. and Karin, Μ., Αηηιι. Rev. Immunol. , 2009, 27: 693-733).

Components of the NF- kappa B pathway, including RIP, IKK and NIK, have been shown to be HSP90 client proteins and inhibition of HSP90 results in degradation of these proteins and blocks activation of the canonical and non-cannonical NF-kappaB pathways (Broemer, M., D. et al., Oncogene, 2004, 23(31 ): 5378-86; Lewis, J. et al., ./. Biol. Chem. , 2000, 275( 14): 10519-26; and Qing, G. and Xiao, G., ./. Biol. Chem., 2005, 280( 1 1 ): 9765-8).

MAP kinases, such as p38 and JNK, have been shown to play key regulatory roles in the production of pro-inflammatory cytokines and downstream signaling events leading to joint inflammation and destruction in arthritis (Thalhamer, T. et al., Rheumatology (Oxford), 2008, 47(4): 409- 14). Inhibition of HSP90 leads to deactivation and degradation of these HSP90 client proteins (Vasilevskaya, I. A. et al., Mol. Pharmacol., 2004, 65( 1 ): 235-43; Salehi, A.H. et al., Chem. Biol. , 2006, 13(2): 213-23; and Wax, S. et al., Arthritis Rheum. , 2003, 48(2): 541 -50). Regulation of mast cell function by the tyrosine kinase KIT is also sensitive to HSP90 inhibition (Fumo, G. et al., Blood, 2004, 103(3): 1078-84).

HSP90 plays a role in the function of both the innate and adaptive immune systems. Data indicate that HSP90 alpha and beta are important for MHC class I and II presentation (Houlihan, J. L. et al., J. Immunol. , 2009, 182( 12): 7451 -7458; Kunisawa, J. and Shastri, N., Immunity, 2006, 24(5): p. 523-34). With respect to the innate immune system, GRP94, which is elevated in rheumatoid arthritis (Huang, Q. Q. et al., J.

Immunol., 2009, 182(8): 4965-73), is a chaperone for Toll-like receptors and is important for the innate function of macrophages (Yang, Y. et al., Immunity, 2007, 26(2): 215-26). Furthermore, inhibition of HSP90 blocks interleukin- 1 receptor associated kinase mediated Toll-like receptor signaling (De Nardo, D. et al., J. Biol. Chem. , 2005, 280( 1 1 ): 9813-22).

Several studies provide in vivo support for the role of HSP90 in inflammation and autoimmune disease. Pharmacological inhibition of HSP90 has been effective in preclinical models of sepsis, multiple sclerosis and uveitis (Dello Russo, C. et al., J.

Neurochem. , 2006, 99(5): 1351 -62; Poulaki, V. et al., Faseh. J., 2007, 21 (9): 21 13-23;

Chatterjee, A. et al., Am. J. Respir. Crit. Care Med., 2007, 176(7): 667-675; and Sugita, T. et al., Biochem. Mol. Biol. Int., 1999, 47(4): 587-95).

The induction of HSP70 has been implicated in inflammatory bowel disease (IBD). Compounds inducing HSP70, such as inhibitors of HSP90, may be useful in the prevention and treatment of inflammatory bowel diseases (IBD) such as Crohn's disease and colitis (see Hu, S.; et al Am. J. Physiol. Gastrointest. Liver Physiol. (2009) G 1003- C 101 1 ; and Tanaka, K-T, et al J. Biol. Chem. (2007) 282; 32, 23240-23252). HSP90 in Cancer

HSP90 has been closely investigated as a target in oncology due to it's central role in the function of key protein networks that become dysregulated in cancer (for reviews see Pearl, L. H., Curr. Opin. Genet. Dev. , 2005, 15( 1 ): 55-61 ; Neckers, L., Trends Mol. Med. , 2002, 8(4 Suppl): S55-61 ; Arslan, M.A. et al., Curr. Cancer Drug Targets, 2006, 6(7): 623-34; and Li, Y. et al., Drug resistance updates : reviews and commentaries in antimicrobial and anticancer chemotherapy, 2009, 12( 1 ): 17-27). In tumor cells, HSP90 is required for pathways that promote cell survival and proliferation. HSP90 regulates the function of oncogenic protein kinases such as AKT, ErbB2, Cdk4 and B-raf . Inhibition of HSP90 results in rapid dephosphorylation and inactivation of kinases such as AKT leading to cancer cell death (Georgakis, G. V. et al., Clin. Cancer Res. , 2006, 12(2): 584- 90). Cytotoxicity studies have demonstrated that HSP90 inhibitors have anti-cancer properties against a wide range of tumor cell lines. HSP90 inhibitors have been shown to be effective in multiple preclinical xenograft cancer models (Bao, R. et al., Clin. Cancer Res., 2009, 15( 12): 4046-4057; Chandarlapaty, S. et al., Clin. Cancer Res., 2008, 14( 1 ): 240-248; Vilenchik, M. et al., Chem. Biol., 2004, 1 1 (6): 787-97; Yin, X. et al., Int. J. Cancer, 2009, Chem. Biol., 2004, 1 1 (6): 787-97; and Leow, C. C. et al., Mol. Cancer Titer., 2009, 8(8): 2131 -41 ). The geldanamycin-related ansamycins, 17-AAG and 17- DMAG, have been tested in clinical trials for the treatment of cancer (Ramanathan, R. K. et al., Clin. Cancer Res., 2005, 1 1 (9): 3385-91 ; Goetz, M. P. et al., ./. Clin. Oncol , 2005, 23(6): 1078-87; and Ronnen, E. A. et al., Invest. New Drugs, 2006, 24(6): 543-6).

Neurology

Inhibition of HSP90 and induction of heat shock proteins is of interest in the treatment of numerous other pathologies. HSP90 inhibitors may have utility in CNS disease. Inhibition of HSP90 has been shown to protect neurons from amyloid beta- induced toxicity (Ansar, S. et al., Bioorg. Med. Chem. Letters, 2007, 17(7): 1984- 1990) as well as reducing the extent of aberrant mutant Tau phosphorylation in neurons (Luo, W. et al., Proc. Natl. Acad. Sci. U S A, 2007, 104(22): 951 1 -6); which suggests that HSP90 inhibitors may be useful in the treatment of Alzheimer's disease. In Parkinson' s disease, HSP90 inhibition may be active through the modulation of disease-related proteins such as alpha-synuclein and LRRK2 (Putcha, P. et al. JPET 2010, 332:849-857; Hurtado- Lorenzo, A. et al. J Neurosci. 2008, 28:6757-6759). HSP90 inhibition has been shown to decrease the release of inflammatory signals from microglia suggesting a possible therapeutic benefit in diseases characterized by neuroinflammation such as multiple sclerosis, stroke, Parkinson' s disease, Alzheimer' s disease, Huntington' s disease and ALS (Dello Russo et al. Neurochem., 2006, 99: 1351 - 1362). HSP90 inhibition may be useful in treatment of stroke; Geldanamycin, a prototypic HSP90 inhibitor, has been shown to be protective in rat models of focal cerebral ischemia. Geldanamycin is also neuroprotective in a cellular model of Amyotropic Lateral Sclerosis (Batulan, Z. et al., Neurobiol. Dis. , 2006, 24(2): 213-25). Additionally, improved cellular processing of peripheral myelin protein 22 in response to HSP90 inhibitors suggests that HSP90 inhibition may be useful in the treatment of hereditary demyelinating neuropathies such as Charcot-Marie-Tooth disease (Rangaraju, S. et al., Neurobiology of Disease, 2008, 32( 1 ): 105- 1 15). HSP90 has also been implicated in retinitis pigmentosa (Tam et al. Hum. Mol. Gen, 2010).

Other indications

In vitro data suggests that HSP90 may be a useful target in the treatment of viral and parasitic infections including rotavirus (Dutta, D. et al., Virology, 2009, 391 (2): 325- 33), influenza (Chase, G. et al., Virology, 2008, 377(2): 431 -9), HCV (Nakagawa, S. et al., Biochem. Biophys. Res. Commun. , 2007, 353(4): 882-8) as well as plasmodium falciparum (Kumar, R. et al., Malar. J., 2003. 2: 30) and filarial nematodes (Devaney, E. et al., Int. ./. Parasitol , 2005, 35(6): 627-36).

Preclinical data demonstrates that inhibition of fungal HSP90 may be

therapeutically useful for the treatment of fungal infections (Bastididas, RJ. et al Cur. Opin. Investig. Drugs, 2008, 9(8): 856-864).

Summary of the Invention

In certain embodiments, the invention relates to compounds having a structure of Formula 1 , or a pharmaceutically acceptable salt thereof

wherein A is selected from Al

each X is independently selected from CR and N, preferably selected such that no more than two occurrences, and more preferably no more than one occurrence, of X are N;

R is selected from -CN and C(0)NH ₂ ;

R ¹ is NH ₂ ;

each R is independently selected from hydrogen, halogen, -N0 ₂ , -CN, alkyl, alkenyl, alkynyl, -OR ³ , -NR ⁴ R ⁵ , -S(0) _m R ³ , -C(0)R ³ , -C(0)OR ³ , -C(0)NR ⁴ R ⁵ , - S(0) ₂ NR ⁴ R ^' \ aryl, heteroaryl, carbocyclyl, and heterocyclyl;

each R', R ⁴ , and R is independently selected from hydrogen, -alkyl-R ⁶ , carbocyclyl, heterocyclyl, heteroaryl, and aryl; or

R ⁴ and R together are alkylene, thereby forming a ring;

R ⁶ is selected from hydrogen, hydroxy, alkoxy, -NHC(0)alkyl, -NHS0 ₂ alkyl, amino, and heterocyclyl;

m is an integer from 0 to 2;

D is selected from heteroaryl, heterocyclyl, and carbocyclyl; and

E is selected from carbocyclyl and heterocyclyl.

In certain embodiments, the invention relates to pharmaceutical compositions comprising a compound of Formula 1 and a pharmaceutically acceptable carrier or diluent.

In certain embodiments, the invention relates to methods for the treatment of a disease or condition selected from autoimmune and inflammatory diseases (e.g., arthritis, multiple sclerosis, lupus and uveitis), sepsis, cancer, neurological diseases (e.g., Alzheimer' s disease, Huntington's disease, Parkinson's disease, Amyotrophic Lateral Sclerosis (ALS), Gerstmann-Straussler-Scheinker syndrome, fatal familial insomnia, Kuru, Creutzfeldt-Jakob disease (CJD), peripheral neuropathies and Charcot-Marie-Tooth disease), inflammatory bowel disease (IBD) such as Crohn's disease and colitis, viral infection (e.g., rotavirus, influenza, and hepatitis C), and parasitic infections (e.g.,

Plasmodium falciparum and filarial nematodes) comprising administering a compound of Formula 1.

Detailed Description of the Invention

In certain embodiments, the invention relates to compounds having a structure of Formula 1 , or a pharmaceutically acceptable salt thereof

wherein

A is selected from A 1 (wherein 1 -6 and 1 -8, respectively refer to positions on the ring);

each X is independently selected from Cir and N;

R is selected from -CN and -C(0)NH ₂ ;

R ¹ is NH ₂ ;

each R ² is independently selected from hydrogen, halogen, -N0 ₂ , -CN, alkyl, alkenyl, alkynyl, -OR ³ , -NR ⁴ R ⁵ , -S(0) _m R ³ , -C(0)R ³ , -C(0)OR ³ , -C(0)NR ⁴ R ⁵ , - S(0) ₂ NR ⁴ R ^' \ aryl, heteroaryl, carbocyclyl, and heterocyclyl;

each R', R ⁴ , and R is independently selected from hydrogen, -alkyl-R ⁶ , carbocyclyl, heterocyclyl, heteroaryl, and aryl; or

R ⁴ and R together are alkylene, thereby forming a ring;

R ⁶ is selected from hydrogen, hydroxy, alkoxy, -NHC(0)alkyl, -NHSC^alkyl, amino, and hetero v lvl : m is an integer from 0 to 2;

D is selected from heteroaryl, heterocyclyl, and carbocyclyl; and

E is selected from carbocyclyl and heterocyclyl.

In certain embodiments, A is Al , wherein each occurrence of X is independently

CR ²

In certain alternative embodiments, A is Al , wherein one or two occurrences of X are N.

In certain preferred embodiments, wherein A is A l and X' (i.e., X at the 3-

2 2

position of Al ) is CR , R is other than hydrogen.

In certain preferred embodiments, X' is CR ² wherein R ² is -NR ⁴ R\ R ⁴ is hydrogen and R is carbocyclyl or heterocyclyl.

In certain preferred embodiments, X' is CR ² wherein R ² is -NR ⁴ R\ R ⁴ is hydrogen and R is heterocyclyl.

In certain preferred embodiments, X ' is CR ² wherein R ² is -NR ⁴ R\ R ⁴ is hydrogen and R is carbocyclyl-R ⁷ wherein

R ⁷ is selected from -OR ¹⁴ , -NR ¹² R ^{1 3} , -C(0)NR ¹² R ¹³ , -S0 ₂ NR ¹² R ¹³ ,

-NR ¹⁴ C(0)R ¹⁵ , -NR ¹⁴ S(0) ₂ R ¹⁵ , -NR ¹⁴ C(0)NR ¹² R ¹³ , -OC(0)NR ¹² R ¹³ , -NR ¹⁴ C(0)OR ¹⁵ , -C(0)OR ¹⁵ , -OC(0)R ¹⁵ or heterocyclyl;

each R ¹² and R ¹ ' is independently selected from hydrogen, alkyl, carbocyclyl, heterocyclyl, aryl and heteroaryl; or

R ¹² and R ¹ ' together form a substituted or unsubstituted heterocyclyl ring system; and

R and R ' are selected from hydrogen, alkyl, carbocyclyl, heterocyclyl, aryl and heteroaryl.

In preferred embodiments, the carbocyclyl group of carbocyclyl-R is a trans 1 -4 disubstituted 6-membered ring.

In certain embodiments as described above, the carbocyclyl group of carbocyclyl-

7 7

R is a six-membered ring. In certain such embodiments, R is preferably located at the 4- position of the carbocyclyl ring relative to the point of attachment to A. In certain embodiments as described above where X' is CR ² , wherein R ² is - NR ⁴ R\ R ⁴ is hydrogen and R is carbocyclyl-R ⁷ , other occurrences of R ² in Formula 1 are independently selected from hydrogen or halogen.

In certain embodiments as described above where X' is CR ² , wherein R ² is - NR ⁴ R\ R ⁴ is hydrogen and R is heterocyclyl, other occurrences of R ² in Formula 1 are independently selected from hydrogen or halogen.

In certain embodiments, A is selected from

In certain embodiments, A is A2, wherein each occurrence of X is independently

CR ² .

In certain embodiments, A is A2, wherein one or two occurrences of X are N. In certain preferred embodiments, R ² when X ⁶ (i.e., X at the 6-position of A2), X ⁷

8 2

(i.e., X at the 7-position of A2) or X (i.e., X at the 8-position of A2) of A2 is CR ,

2 8 2

preferably R when X is CR , is other than hydrogen.

In certain embodiments A is selected from

In certain embodiments, R ² is selected from hydrogen, halogen, -CN, alkyl, -OR', -NR ⁴ R\ -SCOj _m R', and -C(0)NR ⁴ R\ In certain such embodiments, R ² is selected from hydrogen and NR ⁴ R\

In certain embodiments wherein A is Al and X' is CR ² , R ² is selected from In certain embodiments wherein A is Al and X' is CR ² , R ² is selected from

In certain embodiments wherein A is Al and X' is CR ² , R ² is selected from

In certain embodiments wherein A is Al and X' is CR ² , R ² is selected from

In certain embodiments wherein A is Al and X' is CR ² , R ² is selected from

In certain embodiments wherein A is Al and X' is CR ² , R ² is selected from n certain embodiments wherein A is Al and X' is CR ² , R ² is selected from

In certain embodiments wherein A is Al and X' is CR ² , R ² is selected from

In certain embodiments wherein A is A 1 and X '

In certain embodiments wherein A is Al and X' is CR ² , R ² is selected from

In certain embodiments wherein A is Al and X' is CR ² , R ² is selected from

In certain embodiments wherein A is Al and X' is CR ² , R ² is selected from

In certain embodiments wherein A is Al and X ³ is CR ² , R ² is selected irom

In certain embodiments, when A is A2, at least one occurrence of R ² , preferably R ² when

X 6", X 7'or X 8° is CR 2% more preferably R 2 when X 8 is CR 2 , is selected from alkyl, halogen, OR', and -S(0) _m R\ In certain such embodiments, R ² is alkyl, preferably methyl, ethyl, propyl, isopropyl, butyl, or isobutyl. In certain alternative such embodiments, at least one occurrence of R is halogen, preferably Br or F. In certain alternative such embodiments, at least one occurrence of R ² are -OR'. In certain such embodiments, R ² is selected from -0(CH ₂ ) ₂ OCH ₃ , -0(CH ₂ )2N(CH ₃ )2, -0(CH ₂ ) ₂ OH, and OEt. In certain alternative such embodiments, at least one occurrence of R ² is -S(0) _m R\ In certain such embodiments, R ² is selected from

In certain embodiments, B is

wherein Q is selected from -C(O)-, and -S(0) _n -; and n is an integer from 1 to 2. In certain embodiments D is heteroaryl, preferably pyridyl, thienyl, thiazolyl, oxazole, or isoxazole. In certain other embodiments D is carbocyclyl, preferably cyclopentyl or cyclohexyl. In certain embodiments D is heterocyclyl.

In certain embodiments, B is selected from

In certain embodiments, B is selected from

In certain embodiments, B is selected from

In certain embodiments, B is selected from

In certain embodiments, B is selected such that the D ring is one of the D rings specified in the previous paragraph and the E ring is one of the E rings specified in the paragraph prior to that.

Another aspect of the present invention provides a synthetic intermediate compound represented by Formula 2:

wherein X is halogen.

Another aspect of the present invention provides a synthetic intermediate compound represented by Formula 3:

Another aspect of the present invention provides a synthetic intermediate compound represented by Formula 4:

Another aspect of the present invention provides a pharmaceutical composition comprising a compound of Formula 1 and a pharmaceutically acceptable carrier, diluent or excipient.

Another aspect of the present invention provides a method of treating a proliferative disorder or a disease state, the method comprising administering to a subject in need thereof an amount of a compound or pharmaceutical composition as described above sufficient to treat the proliferative disorder or disease state.

Another aspect of the present invention provides a method of modulating HSP function, the method comprising contacting a cell with a compound of the present invention in an amount sufficient to modulate the binding of a HSP client protein to HSP90, thereby modulating the HSP function.

Another aspect of the present invention provides a method of modulating HSP function, the method comprising a) contacting a cell with a compound of the present invention in an amount sufficient to modulate HSP90 function, thereby b) affecting protein folding, stability and aggregation.

Another aspect of the present invention provides a probe, the probe comprising a compound of Formula 1 labeled with a detectable label or an affinity tag. In other words, the probe comprises a residue of a compound of Formula 1 covalently conjugated to a detectable label. Such detectable labels include, but are not limited to, a fluorescent moiety, a chemiluminescent moiety, a paramagnetic contrast agent, a metal chelate, a radioactive isotope-containing moiety, or biotin. As used herein, the term "affinity tag" means a ligand or group, linked either to a compound of the present invention or to an HSP domain, that allows the conjugate to be extracted from a solution.

As used herein, the term "probe" means a compound of the invention which is labeled with either a detectable label or an affinity tag, and which is capable of binding, either covalently or non-covalently, to an HSP domain. When, for example, the probe is non-covalently bound, it may be displaced by a test compound. When, for example, the probe is bound covalently, it may be used to form cross-linked adducts, which may be quantified and inhibited by a test compound.

Uses

In certain embodiments, the invention relates to methods for the treatment of a disease or condition selected from autoimmune diseases and inflammation (e.g., arthritis, multiple sclerosis, lupus, and uveitis), sepsis, cancer, neurological diseases (including neurodegenerative diseases, such as Alzheimer's disease, Huntington' s disease,

Parkinson' s disease, Amyotrophic Lateral Sclerosis (ALS), Gerstmann-Straussler-

Scheinker syndrome, fatal familial insomnia, peripheral neuropathies and Charcot-Marie- Tooth disease, and prion-related diseases such as Kuru, Creutzfeldt-Jakob disease (CJD), and bovine spongiform encephalopathy), inflammatory bowel disease (IBD) such as Crohn's disease and colitis, viral infection (e.g., rotavirus, influenza, and hepatitis C), and parasitic infections (e.g., plasmodium falciparum and filarial nematodes) comprising administering a compound of Formula 1 .

In certain embodiments, the invention relates to a method for treating cancer comprising administering a compound of Formula 1 . In certain such embodiments, the cancer is selected from

Combination Therapy

Another aspect of the invention provides a conjoint therapy wherein one or more other therapeutic agents are therapies are administered with compounds as described herein. Such conjoint treatment may be achieved by way of the simultaneous, sequential, or separate dosing of the individual components of the treatment.

The present invention also relates to the use of the compounds of the present invention in combination with radiation therapy and/or one or more additional agents such as those described in WO 03/09921 1 , which is hereby incorporated by reference. Examples of such additional agents include, but are not limited to the following:

a) an estrogen receptor modulator; b) an androgen receptor modulator; c) a retinoid receptor modulator; d) a cytotoxic agent; e) an antiproliferative agent; f) a prenyl-protein transferase inhibitor; g) an HMG-CoA reductase inhibitor; h) an HIV protease inhibitor; i) a reverse transcriptase inhibitor; j) an angiogenesis inhibitor; k) a PPAR-γ agonist; 1) a ΡΡΑΡν-δ agonist; m) an inhibitor of inherent multidrug resistance; n) an anti-emetic agent; o) an agent useful in the treatment of anemia; p) an agent for the treatment of neutropenia; q) an immunologic-enhancing drug; r) a proteasome inhibitor (e.g., Velcade or MG 132); s) an HDAC inhibitor (e.g., sodium butyrate, phenyl butyrate, hydroxamic acids, cyclin tetrapeptide and the like); t) an inhibitor of the chymotrypsin-like activity in the proteasome; u) an E3 ligase inhibitor; v) a modulator of the immune system (e.g., interferon-alpha and ionizing radiation (UVB) that can induce the release of cytokines, such as the interleukins, TNF, or induce release of Death receptor Ligands, such as TRAIL); w) a modulator of death receptors, including, but not limited to, TRAIL and TRAIL receptor agonists (e.g., humanized antibodies HGS-ETR1 and HGS-ETR2); x) acetylcholinesterase inhibitors; and y) NMD A receptor antagonists.

Additional combinations may also include agents which reduce the toxicity of the aforesaid agents, such as hepatic toxicity, neuronal toxicity, nephrotoxicity and the like. Vinca Alkaloids and Related Compounds

Vinca alkaloids that can be used in combination with the compounds of the invention to treat cancer and other neoplasms include, but are not limited to, vincristine, vinblastine, vindesine, vinflunine, vinorelbine, and anhydrovinblastine.

Taxanes and Other Microtubule Stabilizing Compounds

Taxanes, including, but not limited to, paclitaxel, doxetaxel, RPR 109881 A, SB- T- 1213, SB-T- 1250, SB-T- 101 187, BMS-275183, BRT 216, DJ-927, MAC-321 , IDN5109, and IDN5390, may be used in combination with the compounds of the invention to treat cancer and other neoplasms. Taxane analogs (e.g., BMS- 184476, BMS- 188797) and functionally related non-taxanes (e.g., epothilones (e.g., epothilone A, epothilone B (EPO906), deoxyepothilone B, and epothilone B lactam (BMS-247550)), eleutherobin, discodermolide, 2-epi-discodermolide, 2-des-methyldiscodermolide, 5- hydroxymethyldiscodermolide, 19-des-aminocarbonyldiscodermolide, 9( 13)- cyclodiscodermolide, and laulimalide) can also be used in the methods and compositions of the invention.

Other microtubule stabilizing compounds that can be used in combination with the compounds of the invention to treat cancer and other neoplasms are described in U.S. Pat.

Nos. 6,624,317; 6,610,736; 6,605,599; 6,589,968; 6,583,290; 6,576,658; 6,515,017;

6,531 ,497; 6,500,858; 6,498,257; 6,495,594; 6,489,314; 6,458,976; 6,441 , 186; 6,441 ,025; 6,414,015; 6,387,927; 6,380,395; 6,380,394; 6,362,217; 6,359, 140; 6,306,893; 6,302,838;

6,300,355; 6,291 ,690; 6,291 ,684; 6,268,381 ; 6,262, 107; 6,262,094; 6, 147,234; 6, 136,808;

6, 127,406; 6, 100,41 1 ; 6,096,909; 6,025,385; 6,01 1 ,056; 5,965,718; 5,955,489; 5,919,815;

5,912,263; 5,840,750; 5,821 ,263; 5,767,297; 5,728,725; 5,721 ,268; 5,719, 177; 5,714,513;

5,587,489; 5,473,057; 5,407,674; 5,250,722; 5,010,099; and 4,939, 168; and U.S. patent application Publication Nos. 2003/0186965 Al ; 2003/0176710 Al ; 2003/0176473 Al ;

2003/0144523 Al ; 2003/0134883 Al ; 2003/0087888 Al ; 2003/0060623 Al ;

2003/004571 1 Al ; 2003/0023082 Al ; 2002/0198256 Al ; 2002/0193361 Al ;

2002/0188014 Al ; 2002/0165257 Al ; 2002/01561 10 Al ; 2002/0128471 Al ;

2002/0045609 Al ; 2002/0022651 Al ; 2002/0016356 Al ; 2002/0002292 Al , each of which is hereby incorporated by reference for the compounds disclosed therein.

Other chemotherapeutic agents that may be administered with a compound of the present invention are listed in the following Table: Additional combinations may also include agents which reduce the toxicity of the aforesaid agents, such as hepatic toxicity, neuronal toxicity, nephrotoxicity and the like.

Additional combinations may be used in the treatment of RA such as non-steroidal anti-inflammatory drugs (NSAIDs), analgesics, corticosteroids and disease-modifying antirheumatic drugs. Further combinations may include Kineret, Actemra,

Hydroxychloroquine (Plaquenil™), Sulfasalazine (Azulfidine™), Leflunomide

(Arava™), Tumor Necrosis Factor Inhibitors such as etanercept (Enbrel™), adalimumab (Humira™), and infliximab (Remicade™), T-cell costimulatory blocking agents such as abatacept (Orencia™), B cell depleting agents such as rituximab (Rituxan™), Interleukin- 1 (IL- 1 ) receptor antagonist therapy such as anakinra (Kineret™), intramuscular gold and other immunomodulatory and cytotoxic agents such as azathioprine (Imuran™), cyclophosphamide and cyclosporine A (Neoral™, Sandimmune™).

Other cotherapies for the treatment of RA include Methotrexate, Campath (alemtuzumab), anti-RANKL MAb (denosumab), anti-Blys MAb LymphoStat-B™ (belimumab), Cimzia (certolizumab pegol), p38 inhibitors, JAK inhibitors, SYK inhibitors, ERK inhibitors, FMS inhibitors, cKIT inhibitors, anti-TNF agents, anti-CD2() MAbs, anti-IL/ILR targeting agents such as those which target IL- 1 , IL-5, IL-6

(toclizumab), 11-4, IL- 13, and IL-23.

Additional combinations may be used in the treatment of MS such as Remicade™, Enbrel™, Humira™, Kineret™, Orencia™, Rituxan™ and TYSABRI™ (natalizumab) and Copaxone™ (glatiramer acetate).

i) Administration

Compounds prepared as described herein can be administered in various forms, depending on the disorder to be treated and the age, condition, and body weight of the patient, as is well known in the art. For example, where the compounds are to be administered orally, they may be formulated as tablets, capsules, granules, powders, or syrups; or for parenteral administration, they may be formulated as injections

(intravenous, intramuscular, or subcutaneous), drop infusion preparations, or

suppositories. For application by the ophthalmic mucous membrane route, they may be formulated as eye drops or eye ointments. These formulations can be prepared by conventional means, and if desired, the active ingredient may be mixed with any conventional additive or excipient, such as a binder, a disintegrating agent, a lubricant, a corrigent, a solubilizing agent, a suspension aid, an emulsifying agent, a coating agent, a cyclodextrin, and/or a buffer. Although the dosage will vary depending on the symptoms, age and body weight of the patient, the nature and severity of the disorder to be treated or prevented, the route of administration and the form of the drug, in general, a daily dosage of from 0.01 to 2000 mg of the compound is recommended for an adult human patient, and this may be administered in a single dose or in divided doses. The amount of active ingredient which can be combined with a carrier material to produce a single dosage form will generally be that amount of the compound which produces a therapeutic effect.

The precise time of administration and/or amount of the composition that will yield the most effective results in terms of efficacy of treatment in a given patient will depend upon the activity, pharmacokinetics, and bioavailability of a particular compound, physiological condition of the patient (including age, sex, disease type and stage, general physical condition, responsiveness to a given dosage, and type of medication), route of administration, etc. However, the above guidelines can be used as the basis for fine- tuning the treatment, e.g., determining the optimum time and/or amount of administration, which will require no more than routine experimentation consisting of monitoring the subject and adjusting the dosage and/or timing.

The phrase "pharmaceutically acceptable" is employed herein to refer to those ligands, materials, compositions, and/or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit/risk ratio.

The phrase "pharmaceutically acceptable carrier" as used herein means a pharmaceutically acceptable material, composition, or vehicle, such as a liquid or solid filler, diluent, excipient, solvent or encapsulating material. Each carrier must be

"acceptable" in the sense of being compatible with the other ingredients of the formulation and not injurious to the patient. Some examples of materials which can serve as pharmaceutically acceptable carriers include: ( 1 ) sugars, such as lactose, glucose, and sucrose; (2) starches, such as corn starch, potato starch, and substituted or unsubstituted β-cyclodextrin; (3) cellulose, and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose, and cellulose acetate; (4) powdered tragacanth; (5) malt; (6) gelatin; (7) talc; (8) excipients, such as cocoa butter and suppository waxes; (9) oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and soybean oil; ( 10) glycols, such as propylene glycol; ( 1 1 ) polyols, such as glycerin, sorbitol, mannitol, and polyethylene glycol; ( 12) esters, such as ethyl oleate and ethyl laurate; ( 13) agar; ( 14) buffering agents, such as magnesium hydroxide and aluminum hydroxide; ( 15) alginic acid; ( 16) pyrogen-free water; ( 17) isotonic saline; ( 18) Ringer's solution; ( 19) ethyl alcohol; (20) phosphate buffer solutions; and (21 ) other non-toxic compatible substances employed in pharmaceutical formulations. In certain embodiments, pharmaceutical compositions of the present invention are non-pyrogenic, i.e., do not induce significant temperature elevations when administered to a patient.

The term "pharmaceutically acceptable salt" refers to the relatively non-toxic, inorganic and organic acid addition salts of the compound(s). These salts can be prepared in situ during the final isolation and purification of the compound(s), or by separately reacting a purified compound(s) in its free base form with a suitable organic or inorganic acid, and isolating the salt thus formed. Representative salts include the hydrobromide, hydrochloride, sulfate, bisulfate, phosphate, nitrate, acetate, valerate, oleate, palmitate, stearate, laurate, benzoate, lactate, phosphate, tosylate, citrate, maleate, fumarate, succinate, tartrate, naphthylate, mesylate, glucoheptonate, lactobionate, laurylsulphonate salts, and amino acid salts, and the like. (See, for example, Berge et al. ( 1 77)

"Pharmaceutical Salts", J Pharm. Sci. 66: 1 - 19.)

In other cases, the compounds useful in the methods of the present invention may contain one or more acidic functional groups and, thus, are capable of forming pharmaceutically acceptable salts with pharmaceutically acceptable bases. The term "pharmaceutically acceptable salts" in these instances refers to the relatively non-toxic inorganic and organic base addition salts of a compound(s). These salts can likewise be prepared in situ during the final isolation and purification of the compound(s), or by separately reacting the purified compound(s) in its free acid form with a suitable base, such as the hydroxide, carbonate, or bicarbonate of a pharmaceutically acceptable metal cation, with ammonia, or with a pharmaceutically acceptable organic primary, secondary, or tertiary amine. Representative alkali or alkaline earth salts include the lithium, sodium, potassium, calcium, magnesium, and aluminum salts, and the like. Representative organic amines useful for the formation of base addition salts include ethylamine, diethylamine, ethylenediamine, ethanolamine, diethanolamine, piperazine, and the like (see, for example, Berge et al., supra). Wetting agents, emulsifiers, and lubricants, such as sodium lauryl sulfate and magnesium stearate, as well as coloring agents, release agents, coating agents, sweetening, flavoring, and perfuming agents, preservatives and antioxidants can also be present in the compositions.

Examples of pharmaceutically acceptable antioxidants include: ( 1 ) water soluble antioxidants, such as ascorbic acid, cysteine hydrochloride, sodium bisulfate, sodium metabisulfite, sodium sulfite, and the like; (2) oil-soluble antioxidants, such as ascorbyl palmitate, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), lecithin, propyl gallate, alpha-tocopherol, and the like; and (3) metal chelating agents, such as citric acid, ethylenediamine tetraacetic acid (EDTA), sorbitol, tartaric acid, phosphoric acid, and the like.

Formulations suitable for oral administration may be in the form of capsules, cachets, pills, tablets, lozenges (using a flavored basis, usually sucrose and acacia or tragacanth), powders, granules, or as a solution or a suspension in an aqueous or non- aqueous liquid, or as an oil-in-water or water-in-oil liquid emulsion, or as an elixir or syrup, or as pastilles (using an inert matrix, such as gelatin and glycerin, or sucrose and acacia) and/or as mouthwashes, and the like, each containing a predetermined amount of a compound(s) as an active ingredient. A composition may also be administered as a bolus, electuary, or paste.

In solid dosage forms for oral administration (capsules, tablets, pills, dragees, powders, granules, and the like), the active ingredient is mixed with one or more pharmaceutically acceptable carriers, such as sodium citrate or dicalcium phosphate, and/or any of the following: ( 1 ) fillers or extenders, such as starches, cyclodextrins, lactose, sucrose, glucose, mannitol, and/or silicic acid; (2) binders, such as, for example, carboxymethylcellulose, alginates, gelatin, polyvinyl pyrrolidone, sucrose, and/or acacia; (3) humectants, such as glycerol; (4) disintegrating agents, such as agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates, and sodium carbonate; (5) solution retarding agents, such as paraffin; (6) absorption accelerators, such as quaternary ammonium compounds; (7) wetting agents, such as, for example, acetyl alcohol and glycerol monostearate; (8) absorbents, such as kaolin and bentonite clay; (9) lubricants, such a talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, and mixtures thereof; and ( 10) coloring agents. In the case of capsules, tablets, and pills, the pharmaceutical compositions may also comprise buffering agents. Solid compositions of a similar type may also be employed as fillers in soft and hard-filled gelatin capsules using such excipients as lactose or milk sugars, as well as high molecular weight polyethylene glycols, and the like.

A tablet may be made by compression or molding, optionally with one or more accessory ingredients. Compressed tablets may be prepared using binder (for example, gelatin or hydroxypropylmethyl cellulose), lubricant, inert diluent, preservative, disintegrant (for example, sodium starch glycolate or cross-linked sodium carboxymethyl cellulose), surface-active or dispersing agent. Molded tablets may be made by molding in a suitable machine a mixture of the powdered compound(s) moistened with an inert liquid diluent.

Tablets, and other solid dosage forms, such as dragees, capsules, pills, and granules, may optionally be scored or prepared with coatings and shells, such as enteric coatings and other coatings well known in the pharmaceutical-formulating art. They may also be formulated so as to provide slow or controlled release of the active ingredient therein using, for example, hydroxypropylmethyl cellulose in varying proportions to provide the desired release profile, other polymer matrices, liposomes, and/or microspheres. They may be sterilized by, for example, filtration through a bacteria- retaining filter, or by incorporating sterilizing agents in the form of sterile solid compositions which can be dissolved in sterile water, or some other sterile injectable medium immediately before use. These compositions may also optionally contain opacifying agents and may be of a composition that they release the active ingredient(s) only, or preferentially, in a certain portion of the gastrointestinal tract, optionally, in a delayed manner. Examples of embedding compositions which can be used include polymeric substances and waxes. The active ingredient can also be in micro-encapsulated form, if appropriate, with one or more of the above-described excipients.

Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups, and elixirs. In addition to the active ingredient, the liquid dosage forms may contain inert diluents commonly used in the art, such as, for example, water or other solvents, solubilizing agents, and emulsifiers such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1 ,3-butylene glycol, oils (in particular, cottonseed, groundnut, corn, germ, olive, castor, and sesame oils), glycerol, tetrahydrofuryl alcohol, polyethylene glycols, and fatty acid esters of sorbitan, and mixtures thereof. Besides inert diluents, the oral compositions can also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, coloring, perfuming, and preservative agents.

Suspensions, in addition to the active compound(s), may contain suspending agents as, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar-agar and tragacanth, and mixtures thereof.

Formulations for rectal or vaginal administration may be presented as a suppository, which may be prepared by mixing one or more compound(s) with one or more suitable nonirritating excipients or carriers comprising, for example, cocoa butter, polyethylene glycol, a suppository wax or a salicylate, which is solid at room

temperature, but liquid at body temperature and, therefore, will melt in the rectum or vaginal cavity and release the active agent.

Formulations which are suitable for vaginal administration also include pessaries, tampons, creams, gels, pastes, foams, or spray formulations containing such carriers as are known in the art to be appropriate.

Dosage forms for the topical or transdermal administration of a compound(s) include powders, sprays, ointments, pastes, creams, lotions, gels, solutions, patches, and inhalants. The active component may be mixed under sterile conditions with a pharmaceutically acceptable carrier, and with any preservatives, buffers, or propellants which may be required or beneficial.

The ointments, pastes, creams, and gels may contain, in addition to compound(s), excipients, such as animal and vegetable fats, oils, waxes, paraffins, starch, tragacanth, cellulose derivatives, polyethylene glycols, silicones, bentonites, silicic acid, talc, and zinc oxide, or mixtures thereof.

Powders and sprays can contain, in addition to a compound(s), excipients such as lactose, talc, silicic acid, aluminum hydroxide, calcium silicates, and polyamide powder, or mixtures of these substances. Sprays can additionally contain customary propellants, such as chlorofluorohydrocarbons and volatile unsubstituted hydrocarbons, such as butane and propane.

The compound(s) can be alternatively administered by aerosol. This is arromnlished hv nrenarin ^p _an aqueous aerosol, liposomal preparation, or solid particles containing the composition. A nonaqueous (e.g., fluorocarbon propellant) suspension could be used. Sonic nebulizers are preferred because they minimize exposing the agent to shear, which can result in degradation of the compound.

Ordinarily, an aqueous aerosol is made by formulating an aqueous solution or suspension of the agent together with conventional pharmaceutically acceptable carriers and stabilizers. The carriers and stabilizers vary with the requirements of the particular composition, but typically include nonionic surfactants (Tweens, Pluronics, sorbitan esters, lecithin, Cremophors), pharmaceutically acceptable co-solvents such as polyethylene glycol, innocuous proteins like serum albumin, oleic acid, amino acids such as glycine, buffers, salts, sugars, or sugar alcohols. Aerosols generally are prepared from isotonic solutions.

Transdermal patches have the added advantage of providing controlled delivery of a compound(s) to the body. Such dosage forms can be made by dissolving or dispersing the agent in the proper medium. Absorption enhancers can also be used to increase the flux of the compound(s) across the skin. The rate of such flux can be controlled by either providing a rate controlling membrane or dispersing the compound(s) in a polymer matrix or gel.

Pharmaceutical compositions of this invention suitable for parenteral

administration comprise one or more compound(s) in combination with one or more pharmaceutically acceptable sterile aqueous or nonaqueous solutions, dispersions, suspensions or emulsions, or sterile powders which may be reconstituted into sterile injectable solutions or dispersions just prior to use, which may contain antioxidants, buffers, bacteriostats, solutes which render the formulation isotonic with the blood of the intended recipient or suspending or thickening agents.

Examples of suitable aqueous and nonaqueous carriers which may be employed in the pharmaceutical compositions of the invention include water, ethanol, polyols (such as glycerol, propylene glycol, polyethylene glycol, and the like), and suitable mixtures thereof, vegetable oils, such as olive oil, and injectable organic esters, such as ethyl oleate. Proper fluidity can be maintained, for example, by the use of coating materials, such as lecithin, by the maintenance of the required particle size in the case of dispersions, and by the use of surfactants. These compositions may also contain adjuvants such as preservatives, wetting agents, emulsifying agents, and dispersing agents. Prevention of the action of microorganisms may be ensured by the inclusion of various antibacterial and antifungal agents, for example, paraben, chlorobutanol, phenol sorbic acid, and the like. It may also be desirable to include tonicity-adjusting agents, such as sugars, sodium chloride, and the like into the compositions. In addition, prolonged absorption of the injectable pharmaceutical form may be brought about by the inclusion of agents which delay absorption such as aluminum monostearate and gelatin.

In some cases, in order to prolong the effect of a drug, it is desirable to slow the absorption of the drug from subcutaneous or intramuscular injection. For example, delayed absorption of a parenterally administered drug form is accomplished by dissolving or suspending the drug in an oil vehicle.

Injectable depot forms can be made by forming microencapsulated matrices of compound(s) in biodegradable polymers such as polylactide-polyglycolide. Depending on the ratio of drug to polymer, and the nature of the particular polymer employed, the rate of drug release can be controlled. Examples of other biodegradable polymers include poly(orthoesters) and poly(anhydrides). Depot injectable formulations are also prepared by entrapping the drug in liposomes or microemulsions which are compatible with body tissue.

The preparations of agents may be given orally, parenterally, topically, or rectally.

They are, of course, given by forms suitable for each administration route. For example, they are administered in tablets or capsule form, by injection, inhalation, eye lotion, ointment, suppository, infusion; topically by lotion or ointment; and rectally by suppositories. Oral administration is preferred.

The phrases "parenteral administration" and "administered parenterally" as used herein means modes of administration other than enteral and topical administration, usually by injection, and includes, without limitation, intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal,

intraperitoneal, transtracheal, subcutaneous, subcuticular, intraarticular, subcapsular, subarachnoid, intraspinal and intrasternal injection, and infusion.

The phrases "systemic administration," "administered systemically," "peripheral administration" and "administered peripherally" as used herein mean the administration of a ligand, drug, or other material other than directly into the central nervous system, such that it enters the patient' s system and thus, is subject to metabolism and other like processes, for example, subcutaneous administration.

These compound(s) may be administered to humans and other animals for therapy by any suitable route of administration, including orally, nasally, as by, for example, a spray, rectally, intravaginally, parenterally, intracisternally, and topically, as by powders, ointments or drops, including buccally and sublingually.

Regardless of the route of administration selected, the compound(s), which may be used in a suitable hydrated form, and/or the pharmaceutical compositions of the present invention, are formulated into pharmaceutically acceptable dosage forms by conventional methods known to those of skill in the art.

Actual dosage levels of the active ingredients in the pharmaceutical compositions of this invention may be varied so as to obtain an amount of the active ingredient which is effective to achieve the desired therapeutic response for a particular patient, composition, and mode of administration, without being toxic to the patient.

The concentration of a disclosed compound in a pharmaceutically acceptable mixture will vary depending on several factors, including the dosage of the compound to be administered, the pharmacokinetic characteristics of the compound(s) employed, and the route of administration. In general, the compositions of this invention may be provided in an aqueous solution containing about 0.1 - 10% w/v of a compound disclosed herein, among other substances, for parenteral administration. Typical dose ranges are from about 0.01 to about 50 mg/kg of body weight per day, given in 1 -4 divided doses. Each divided dose may contain the same or different compounds of the invention. The dosage will be an effective amount depending on several factors including the overall health of a patient, and the formulation and route of administration of the selected compound(s).

Definitions

As used herein, the term "affinity tag" means a ligand or group, linked either to a compound of the present invention or to an HSP domain, that allows the conjugate to be extracted from a solution. The term "alkyl" refers to substituted or unsubstituted saturated hydrocarbon groups, including straight-chain alkyl and branched-chain alkyl groups, including haloalkyl groups such as trifluoromethyl and 2,2,2-trifluoroethyl, etc. Representative alkyl groups include methyl, ethyl, n-propyl, isopropyl, n-butyl, t-butyl, isobutyl, sec- butyl, (cyclohexyl)methyl, cyclopropylmethyl, n-pentyl, n-hexyl, n-heptyl, n-octyl, and the like. The terms "alkenyl" and "alkynyl" refer to substituted or unsubstituted unsaturated aliphatic groups analogous in length and possible substitution to the alkyls described above, but that contain at least one double or triple bond respectively.

Representative alkenyl groups include vinyl, propen-2-yl, crotyl, isopenten-2-yl, 1 ,3- butadien-2-yl), 2,4-pentadienyl, and 1 ,4-pentadien-3-yl. Representative alkynyl groups include ethynyl, 1 - and 3-propynyl, and 3-butynyl. In certain preferred embodiments, alkyl substituents are lower alkyl groups, e.g., having from 1 to 6 carbon atoms.

Similarly, alkenyl and alkynyl preferably refer to lower alkenyl and alkynyl groups, e.g., having from 2 to 6 carbon atoms. As used herein, "alkylene" refers to an alkyl group with two open valencies (rather than a single valency), such as -(CH ₂ )i_io- and substituted variants thereof.

The term "alkoxy" refers to an alkyl group having an oxygen attached thereto. Representative alkoxy groups include methoxy, ethoxy, propoxy, tert-butoxy and the like. An "ether" is two hydrocarbons covalently linked by an oxygen. Accordingly, the substituent of an alkyl that renders that alkyl an ether is or resembles an alkoxy.

The term "alkoxyalkyl" refers to an alkyl group substituted with an alkoxy group, thereby forming an ether.

The terms "amide" and "amido" are art-recognized as an amino- substituted carbonyl and includes a moiety that can be represented by the general formula:

wherein R ⁹ , R ¹⁰ are as defined above. Preferred embodiments of the amide will not include imides, which may be unstable.

The terms "amine" and "amino" are art-recognized and refer to both unsubstituted and substituted amines and salts thereof, e.g., a moiety that can be represented by the general formulae:

wherein R ⁹ , R ¹⁰ and R ¹⁰ each independently represent a hydrogen, an alkyl, an alkenyl, -(CH ₂ ) _m -R ⁸ , or R ⁹ and R ¹⁰ taken together with the N atom to which they are attached complete a heterocycle having from 4 to 8 atoms in the ring structure; R represents an aryl, a cycloalkyl, a cycloalkenyl, a heterocyclyl or a polycyclyl; and m is zero or an integer from 1 to 8. In preferred embodiments, only one of R or R" can be a carbonyl, e.g., R ⁹ , R ¹⁰ , and the nitrogen together do not form an imide. In even more preferred embodiments, R ⁹ and R ¹⁰ (and optionally R ¹⁰ ) each independently represent a hydrogen, an alkyl, an alkenyl, or -(CH ₂ ) _m -R■ In certain embodiments, the amino group is basic, meaning the protonated form has a pK _a > 7.00.

The term "aralkyl", as used herein, refers to an alkyl group substituted with an aryl group.

The term "aryl" as used herein includes 5-, 6-, and 7-membered substituted or unsubstituted single-ring aromatic groups in which each atom of the ring is carbon. The term "aryl" also includes polycyclic ring systems having two or more cyclic rings in which two or more carbons are common to two adjoining rings wherein at least one of the rings is aromatic, e.g., the other cyclic rings can be cycloalkyls, cycloalkenyls, cycloalkynyls, aryls, heteroaryls, and/or heterocyclyls. Aryl groups include benzene, naphthalene, phenanthrene, phenol, aniline, anthracene, and phenanthrene.

The terms "carbocycle" and "carbocyclyl", as used herein, refer to a non-aromatic substituted or unsubstituted ring in which each atom of the ring is carbon. The terms "carbocycle" and "carbocyclyl" also include polycyclic ring systems having two or more cyclic rings in which two or more carbons are common to two adjoining rings wherein at least one of the rings is carbocyclic, e.g., the other cyclic rings can be cycloalkyls, cycloalkenyls, cycloalkynyls, aryls, heteroaryls, and/or heterocyclyls. Representative carbocyclic groups include cyclopentyl, cyclohexyl, 1 -cyclohexenyl, and 3- cyclohexen- 1 -yl, cycloheptyl.

The term "carbonyl" is art-recognized and includes such moieties as can be represented by the general formula:

wherein X is a bond or represents an oxygen or a sulfur, and R ^{1 1} represents a hydrogen, an alkyl, an alkenyl, -(CH ₂ ) _m - or a pharmaceutically acceptable salt. Where X is an oxygen and R ^{1 1} is not hydrogen, the formula represents an "ester". Where X is an oxygen, and R ^{1 1} is a hydrogen, the formula represents a "carboxylic acid".

The terms "heteroaryl" includes substituted or unsubstituted aromatic 5- to 7- membered ring structures, more preferably 5- to 6-membered rings, whose ring structures include one to four heteroatoms. The term "heteroaryl" also includes polycyclic ring systems having two or more cyclic rings in which two or more carbons are common to two adjoining rings wherein at least one of the rings is heteroaromatic, e.g., the other cyclic rings can be cycloalkyls, cycloalkenyls, cycloalkynyls, aryls, heteroaryls, and/or heterocyclyls. Heteroaryl groups include, for example, pyrrole, furan, thiophene, imidazole, isoxazole, oxazole, thiazole, triazole, pyrazole, pyridine, pyrazine, pyridazine and pyrimidine, and the like.

The term "heteroatom" as used herein means an atom of any element other than carbon or hydrogen. Preferred heteroatoms are nitrogen, oxygen, and sulfur.

The terms "heterocyclyl" or "heterocyclic group" refer to substituted or unsubstituted non-aromatic 3- to 10-membered ring structures, more preferably 3- to 7- membered rings, whose ring structures include one to four heteroatoms. The term terms "heterocyclyl" or "heterocyclic group" also include polycyclic ring systems having two or more cyclic rings in which two or more carbons are common to two adjoining rings wherein at least one of the rings is heterocyclic, e.g., the other cyclic rings can be cycloalkyls, cycloalkenyls, cycloalkynyls, aryls, heteroaryls, and/or heterocyclyls.

Heterocyclyl groups include, for example, tetrahydrofuran, tetrahydropyran, piperidine, piperazine, pyrrolidine, morpholine, thiomorpholine, diazapine, lactones, and lactams.

The term "hydrocarbon", as used herein, refers to a group that is bonded through a carbon atom that does not have a =0 or =S substituent, and typically has at least one carbon-hydrogen bond and a primarily carbon backbone, but may optionally include heteroatoms. Thus, groups like methyl, ethoxyethyl, 2-pyridyl, and trifluoromethyl are considered to be hydrocarbyl for the purposes of this application, but substituents such as acetyl (which has a =0 substituent on the linking carbon) and ethoxy (which is linked through oxygen, not carbon) are not. Hydrocarbyl groups include, but are not limited to aryl, heteroaryl, carbocycle, heterocycle, alkyl, alkenyl, alkynyl, and combinations thereof.

The terms "polycyclyl" or "polycyclic" refer to two or more rings (e.g., cycloalkyls, cycloalkenyls, cycloalkynyls, aryls, heteroaryls, and/or heterocyclyls) in which two or more carbons are common to two adjoining rings, e.g., the rings are "fused rings". Each of the rings of the polycycle can be substituted or unsubstituted.

The term "preventing" is art-recognized, and when used in relation to a condition, such as a local recurrence (e.g., pain), a disease such as cancer, a syndrome complex such as heart failure or any other medical condition, is well understood in the art, and includes administration of a composition which reduces the frequency of, or delays the onset of, symptoms of a medical condition in a subject relative to a subject which does not receive the composition. Thus, prevention of cancer includes, for example, reducing the number of detectable cancerous growths in a population of patients receiving a prophylactic treatment relative to an untreated control population, and/or delaying the appearance of detectable cancerous growths in a treated population versus an untreated control population, e.g., by a statistically and/or clinically significant amount. Prevention of an infection includes, for example, reducing the number of diagnoses of the infection in a treated population versus an untreated control population, and/or delaying the onset of symptoms of the infection in a treated population versus an untreated control population. Prevention of pain includes, for example, reducing the magnitude of, or alternatively delaying, pain sensations experienced by subjects in a treated population versus an untreated control population.

As used herein, the term "probe" means a compound of the invention which is labeled with either a detectable label or an affinity tag, and which is capable of binding, either covalently or non-covalently, to an HSP90 domain. When, for example, the probe is non-covalently bound, it may be displaced by a test compound. When, for example, the probe is bound covalently, it may be used to form cross-linked adducts, which may be quantified and inhibited by a test compound.

The term "substituted" refers to moieties having substituents replacing a hydrogen on one or more carbons of the backbone. It will be understood that "substitution" or "substituted with" includes the implicit proviso that such substitution is in accordance with permitted valence of the substituted atom and the substituent, and that the substitution results in a stable compound, e.g., which does not spontaneously undergo transformation such as by rearrangement, cyclization, elimination, etc. As used herein, the term "substituted" is contemplated to include all permissible substituents of organic compounds. In a broad aspect, the permissible substituents include acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic and non-aromatic substituents of organic compounds. The permissible substituents can be one or more and the same or different for appropriate organic compounds. For purposes of this invention, the heteroatoms such as nitrogen may have hydrogen substituents and/or any permissible substituents of organic compounds described herein which satisfy the valences of the heteroatoms. Substituents can include, for example, a halogen, a hydroxyl, a carbon yl (such as a carboxyl, an alkoxycarbonyl, a formyl, or an acyl), a thiocarbonyl (such as a thioester, a thioacetate, or a thioformate), an alkoxyl, a phosphoryl, a phosphate, a phosphonate, a phosphinate, an amino, an amido, an amidine, an imine, a cyano, a nitro, an azido, a sulfhydryl, an alkylthio, a sulfate, a sulfonate, a sulfamoyl, a sulfonamido, a sulfonyl, a heterocyclyl, an aralkyl, or an aromatic or heteroaromatic moiety. It will be understood by those skilled in the art that the moieties substituted on the hydrocarbon chain can themselves be substituted, if appropriate.

As used herein, the term "treating" or "treatment" includes reversing, reducing, or arresting the symptoms, clinical signs, and underlying pathology of a condition in manner to improve or stabilize a subject's condition.

Compounds of the invention also include all isotopes of atoms present in the intermediates and/or final compounds. Isotopes include those atoms having the same atomic number but different mass numbers. For example, isotopes of hydrogen include deuterium and tritium.

Exemplification

Illustrated below are general synthetic procedures for the preparation of compounds of general formula 1 .

Condensation of intermediate 1 -a, wherein X is halogen, with dione 1 -b provides the enamine intermediate 1 -c. Intramolecular Heck condensation of 1 -c provides intermediate 1 -d. Intermediate 1 -d is treated with a base and 2-bromo-4- fluorobenzonitrile to provide intermediate 1 -e. Buchwald coupling of intermediate 1 -e using a catalyst, a ligand, a base and R R NH provides intermediate 1 -f. Nitrile hydrolysis of intermediate 1 -f provides compounds of formula 1.

Scheme 1

Treatment of commercially available aryl halide 2-a with an amine of formula R R NH provides nitrile intermediate 2-b. Condensation of amino alcohol 2-f with dione 1 -b provides β-hydroxy-enamine intermediate 2-g. Intermediate 2-g is then treated with a catalyst, a base and mesityl bromide to provide intermediate 2-e. intermediate 2-e can be prepared using an alternative approach; oxime intermediate 2-d is prepared from the corresponding β-keto ester 2-c and then treated with dione 1 -b to provide intermediate 2- e. Intermediate 2-b and intermediate 2-e are then treated in the presence of N, N'- dimethylethane- 1 ,2-diamine, a base and copper (I) iodide to provide intermediate 2-d. Nitrile hydrolysis of intermediate 2-d provide compounds of formula 1 .

Scheme 2

Synthesis of Compound 2

Scheme 3

Step 1: 3-b A mixture of 3-bromopyridin-2-amine ( 1 .85 g, 10.69 mmol), 5,5- dimethylcyclohexane- l ,3-dione ( 1 .95 g, 13.90 mmol) and TsOH ( 102 mg, 0.54 mmol) in toluene was refluxed in a Dean-Stark apparatus for 24 hours. The reaction was then cooled to room temperature and concentrated under reduced pressure. Saturated aqueous NaHCO and CH2CI2 were added, the organic layer was separated, the aqueous layer was extracted with CH2CI2, and the combined organic layers were dried over anhydrous MgS0 ₄ , filtered and concentrated in vacuo. Purification by silica gel chromatography provided intermediate 3-b as a white solid. MS (m/z) M+H= 296.8.

Step 2: 3-c

To a solution of intermediate 3-b ( 1.28 g, 4.34 mmol) in anhydrous pyridine were sequentially added Cy ₂ NMe ( 1 .1 1 raL, 5.20 mmol) and Pd(PPh ₃ ) ₄ (251 mg, 0.21 mmol). The reaction mixture was stirred at 150 °C for 30 minutes, cooled to room temperature, diluted with toluene and then concentrated under reduced pressure. The residue was dissolved in CH2CI2, water was added, and the organic layer was separated, dried over anhydrous MgS0 ₄ , filtered and concentrated in vacuo. Purification by silica gel chromatography provided intermediate 3-c as a yellow solid. MS (m/z) M+H= 215.1.

Step 3: 3-d

NaH (54.3 mg, 1 .35 mmol) was added to a solution of intermediate 3-c (291 mg, 1 .35 mmol) in DMF cooled to 0 °C and the mixture was stirred for 5 minutes. 2-bromo- 4-fluorobenzonitrile (380 mg, 1.90 mmol) was added and the reaction was then stirred at 67 °C for 4 hours and then cooled to room temperature. Water and ethyl acetate were added, the organic layer was separated, washed with brine, dried over anhydrous MgSC>4, filtered and concentrated in vacuo. Purification by silica gel chromatography provided intermediate 3-d as a yellow solid. MS (m/z) M+H= 395.8

Step 4: 3-e

To a suspension of intermediate 3-d ( 128 mg, 0.32 mmol) and trans-4- aminocyclohexanol ( 150 mg, 1 .29 mmol) in toluene were added palladium(II) acetate (3.64 mg), l , -bis(diphenylphosphino)ferrocene ( 18.19 mg, 0.03 mmol) and sodium tert- butoxide (62.4 mg, 0.64 mmol). The reaction mixture was heated to 1 15 °C for 1.5 hours. The reaction was cooled to room temperature and diethyl ether was added. The mixture was filtered, the precipitate was washed twice with ethyl acetate and the filtrate was dried over anhydrous MgS0 ₄ , filtered and concentrated in vacuo. Purification by silica gel chromatography provided intermediate 3-e as a yellow solid. MS (m/z) M+H= 429.5

Step 5: Compound 2

To a solution of intermediate 3-e (40 mg, 0.09 mmol) in MeOH ( 1 mL) and DMSO (333 uL) was added NaOH I N (467 μΐ _^ , 0.46 mmol) and 30% hydrogen peroxide (95 μΐ _^ , 0.93 mmol) and the reaction was stirred for 1 hour at room temperature. 10% Na ₂ S ₂ 0 ₃ ( 1 mL) was added, volatiles were removed under reduced pressure and ethyl acetate was added. The organic layer was separated, the aqueous phase was extracted with ethyl acetate, the combined organic layers were dried over anhydrous MgSC>4, filtered and concentrated in vacuo. Purification by silica gel chromatography provided compound 2 as a yellow solid. MS (m/z) M+H= 447.4.

Synthesis of compound 4

Scheme 4

Step 1: 4-b

A mixture of tert-butyl 3-bromothiophen-2-yl carbamate 4-a ( 1.00 g, 3.59 mmol), 5,5-dimethylcyclohexane- l ,3-dione (750 mg, 5.39 mmol) and TsOH ( 124 mg, 0.71 mmol) in toluene was refluxed in a Dean-Stark apparatus for 16 hours. The reaction was then cooled to room temperature, saturated aqueous NaHCC^ and ethyl acetate were added, and the organic layer was separated, dried over anhydrous MgS0 ₄ , filtered and concentrated in vacuo. Purification by silica gel chromatography provided intermediate 4- b as a white solid. MS (m/z) M+H= 301.7

Step 2: 4-c

To a solution of intermediate 4-b (600 mg, 1.99 mmol) in anhydrous toluene were sequentially added Cy ₂ NMe ( 1 .28 mL, 6.00 mmol), tri-tert-butylphosphonium

tetrafluoroborate ( 174 mg, 0.60 mmol), tris(dibenzylideneacetone) dipalladium- chloroform adduct ( 155 mg, 0.15 mmol). The reaction was stirred at 100 °C for 1 hour, cooled to room temperature and diluted with ethyl acetate. Saturated aqueous ammonium chloride was added, the organic layer was separated, washed with brine, dried over anhydrous MgS0 ₄ , filtered and concentrated in vacuo. Purification by silica gel chromatography provided intermediate 4-c as a yellow solid. MS (m/z) M+H= 220.2

Step 3: 4-d

NaH (88 mg, 2.18 mmol) was added to a solution of intermediate 4-c (400 mg,

1 .82 mmol) in DMF cooled to 0 °C and the mixture was stirred for 5 minutes. 2-Bromo- 4-fluorobenzonitrile (51 1 mg, 2.55 mmol) was added and the reaction was then stirred at 50 °C overnight and then cooled to room temperature. Saturated aqueous ammonium chloride and ethyl acetate were added, the organic layer was separated, washed with brine, dried over anhydrous MgSC>4, filtered and concentrated in vacuo. Purification by silica gel chromatography provided intermediate 4-d as a yellow solid.

Step 4: Compound 4

To a suspension of intermediate 4-d (200 mg, 0.50 mmol) and trans-4- aminocyclohexanol (231 mg, 2.00 mmol) in toluene were sequentially added

palladium(II) acetate ( 1 1 mg), 1 , 1 '-Bis(diphenylphosphino)ferrocene (56.0 mg, 0.10 mmol) and sodium tert-butoxide (96 mg, 1.00 mmol), the reaction was heated to 1 15 °C for 5 hours and then cooled to room temperature. Methanol/water 1 : 1 was added and volatiles were removed in vacuo. Purification of the residue by silica gel chromatography provided compound 4 as a yellow solid. MS (m/z) M+H= 452.4 Synthesis of compound 9

Scheme 5

Step 1: 5-b

To a solution of NaOH ( 14.09 g, 352.0 mmol) in water (200 mL) cooled to 0°C were sequentially added ethyl 2-oxocyclopentanecarboxylate (50.0 g, 320.0 mmol) and a solution of sodium nitrite (24.30 g, 352.0 mmol) in water ( 120.0 mL) dropwise over a period of 20 minutes and the reaction mixture was then stirred at room temperature for 4 days. Diethyl ether was added, the organic layer was separated and the aqueous phase was acidified to pH=3 with I N HC1. Diethyl ether was added, the organic layer was separated, the aqueous phase was extracted with diethyl ether, the combined organic extracts were washed with brine, dried over anhydrous MgS0 ₄ , filtered and concentrated in vacuo to provide intermediate 5-b as yellow solid.

Step 2: 5-c

To a solution of 5,5-dimethylcyclohexane- l ,3-dione (26.8 g, 191.0 mmol) and intermediate 5-b (21.6 g, 191.0 mmol) in 70% acetic acid (220 mL) was added zinc dust (37.5 g, 573.0 mmol) portion wise over a period of 30 minutes, the reaction was then stirred at reflux for 2 hours and then cooled to room temperature. Water (400 mL) was added and intermediate 5-c was collected by filtration as a white solid. MS (m/z) M+H=204.1

Step 3: 5-e

A solution of 4-bromo-2,6-difluorobenzonitrile (3.0 g, 13.76 mmol), trans-4- aminocyclohexanol ( 1.58 g, 13.76 mmol) and triethylamine ( 1 .93 mL, 13.76 mmol) in DMSO was stirred at room temperature for 72 hours. Saturated aqueous ammonium chloride and ethyl acetate were added; the organic layer was separated, washed with saturated aqueous ammonium chloride and brine, dried over anhydrous MgSC>4, filtered and concentrated in vacuo. Purification by silica gel chromatography provided intermediate 5-e as a white solid.

Step 4: 5-f

To a solution of intermediate 5-e (616 mg, 1.96 mmol) and intermediate 5-c (400 mg, 1 .96 mmol) in dioxane were sequentially added potassium carbonate ( 1 .36 g, 22.83 mmol), copper (I) iodide (562 mg, 2.95 mmol) and Nl , N2-dimethylethane- 1 ,2-diamine ( 195 L, 1.96 mmol), the reaction was then heated at 100°C overnight and then cooled to room temperature. Ethyl acetate was added; the reaction was filtered through celite and the filtrate was reduced in vacuo. Purification by silica gel chromatography provided intermediate 5-f as a yellow solid. MS (m/z) M+H=436.4

Step 5: Compound 9

To a solution of intermediate 5-f (320 mg, 0.73 mmol) in ethanol (8.40 mL) and DMSO (2.0 mL) were sequentially added NaOH I N (8.82 mL, 8.82 mmol) and 30% aqueous hydrogen peroxide (300 L, 2.94 mmol), the reaction was stirred at 50°C for 4 hours and then cooled to room temperature. Water and ethyl acetate were added; the organic layer was separated, washed with saturated aqueous ammonium chloride and brine, dried over anhydrous MgSC>4, filtered and concentrated in vacuo. Purification by silica gel chromatography provided compound 9 as a white solid. MS (m/z) M+H= 454.4 Synthesis of compound 16

Scheme 6

Step 1: 6-b

A mixture of intermediate 6-a ( 17.97 g, 174.0 mmol) and 5,5- dimethylcyclohexane- l ,3-dione (24.43 g, 174.0 mmol) in toluene was refluxed for 4 hours in a dean stark apparatus and then cooled to room temperature. A precipitate formed, the solid was collected by filtration, washed with toluene and diethyl ether, and dried in vacuo to provide intermediate 6-b as white solid. MS (m/z) M+H=226.1

Step 2: 6-c

To a solution of intermediate 6-b ( 10.0 g, 44.4 mmol) in DMF were sequentially added potassium carbonate ( 12.58 g, 91.0 mmol), Pd(PPh ₃ ) ₄ ( 1 .16 g, 4.44 mmol), triphenylphosphine ( 1.16 g, 4.44 mmol) and mesityl bromide (8.84 g, 44.4 mmol) and the reaction was stirred at 150 °C overnight and then cooled to room temperature. Saturated aqueous ammonium chloride and ethyl acetate were added; the organic layer was separated, washed with saturated aqueous ammonium chloride and brine, dried over anhydrous MgS0 ₄ , filtered and concentrated in vacuo. Purification by silica gel chromatography provided intermediate 6-c as a yellow solid. MS (m/z) M+H=2()6.1

Step 3: 6-d

To a solution of intermediate 5-e (5.20 g, 16.61 mmol) and intermediate 6-c (3.10 g, 15.10 mmol) in dioxane were sequentially added potassium carbonate ( 10.44 g, 76.0 mmol), copper (I) iodide (4.31 g, 22.65 mmol) and Nl , N2-dimethylethane- 1 ,2-diamine (2.1 1 mL, 1 .63 mmol) and the reaction was then heated at 125°C for 2 days and then cooled to room temperature. Saturated aqueous ammonium chloride and ethyl acetate was added; the organic layer was separated, washed with brine, dried over MgS04, filtered and concentrated in vacuo. Purification by silica gel chromatography provided intermediate 6-d as a yellow solid. MS (m/z) M+H= 438.5

Step 4: Compound 16

To a solution of intermediate 6-d (3.60 g, 8.23 mmol) in methanol (82 mL) and DMSO (82 mL) were sequentially added NaOH I N (8.23 mL, 8.23 mmol) and 30% aqueous hydrogen peroxide ( 1.26 mL, 12.34 mmol) and the reaction was stirred at room temperature for 1 hour. Water and ethyl acetate were added; the organic layer was separated, washed with saturated aqueous ammonium chloride and brine, dried over MgSC>4, filtered and concentrated in vacuo. Purification by silica gel chromatography provided compound 16 as a white solid. MS (m/z) M+H= 456.4

Synthesis of compound 17

Scheme 7

Step 1: 7-b

A solution of 2-aminocyclohexanol (4.22 g, 36.6 mmol) and 5,5- dimethylcyclohexane- l ,3-dione (5.14 g, 36.6 mmol) in toluene was stirred under reflux for 2 hours and then cooled to room temperature. A precipitate formed and intermediate 7-b was isolated by filtration as a yellow solid.

Step 2: 7-c

To a solution of intermediate 7-b ( 1.18 g, 4.97 mmol) in DMF were sequentially added potassium carbonate ( 1.40 g, 10.18 mmol), Pd(PPh ₃ ) ₄ ( 144 mg, 0.12 mmol) and mesityl bromide ( 1.0 g, 5.07 mmol) and the reaction was stirred at 150 °C overnight and then cooled to room temperature. Water was added, a precipitate formed and intermediate 7-c was collected by filtration as an off-white solid. MS (m/z) M+H=218.1

Step 3: 7-e A solution of 4-bromo-2-fluorobenzonitrile 7-d (25.0 g, 125.0 mmol), trans-4- aminocyclohexanol ( 14.40 g, 125.0 mmol) and triethylamine (52.3 raL, 375.0 mmol) in DMSO was heated at 150 °C overnight and then cooled to room temperature. Saturated aqueous ammonium chloride and ethyl acetate were added; the organic layer was separated, washed with saturated aqueous ammonium chloride and brine, dried over anhydrous MgS0 ₄ , filtered and concentrated in vacuo. Purification by silica gel chromatography provided intermediate 7-e as a white solid.

Step 4: 7-f

To a solution of intermediate 7-e (244 mg, 0.82 mmol) and intermediate 7-c ( 150 mg, 0.69 mmol) in 1 ,4-dioxane were sequentially added potassium carbonate (572 mg, 4.14 mmol), copper (I) iodide (237 mg, 1.24 mmol) and Nl , N2-dimethylethane- l ,2- diamine ( 195 L, 1 .96 mmol) and the reaction was then heated at 100°C overnight and then cooled to room temperature. Ethyl acetate was added; the reaction was filtered through celite and the filtrate was reduced in vacuo. Purification by silica gel chromatography provided intermediate 7-f as a yellow solid. MS (m/z) M+H= 432.6

Step 5: Compound 17

To a solution of intermediate 7-f (320 mg, 0.73 mmol) in ethanol ( 12 raL) and DMSO (3.0 raL) were sequentially added NaOH I N ( 12.0 raL, 12.0 mmol) and 30% aqueous hydrogen peroxide (8.0 mL, 78.0 mmol) and the reaction was stirred at room temperature for 2 hours. Water was added, a precipitate formed and compound 17 was isolated by filtration as a white solid. MS (m/z) M+H= 450.5

Synthesis of compound 11

Scheme 8

Step 1: 8-a

A solution of 4-bromo-2,6-difluorobenzonitrile 5-d ( 1.17 g, 5.39 mmol), 4- aminotetrahydro-2H-thiopyran 1 , 1 -dioxyde ( 1.17 g, 5.39 mmol) and TEA ( 1.87 mL, 13.46 mmol) in DMSO was stirred at room temperature overnight. Saturated aqueous ammonium chloride and ethyl acetate were added; the organic layer was separated, washed with saturated aqueous ammonium chloride and brine, dried over anhydrous MgS0 ₄ , filtered and concentrated in vacuo. Purification by silica gel chromatography provided intermediate 8-a as a white solid.

Step 2: 8-b

To a solution of intermediate 8-a (307 mg, 0.88 mmol) and intermediate 5-c ( 150 mg, 0.74 mmol) in dioxane were sequentially added potassium carbonate (510 mg, 3.69 mmol), copper (I) iodide ( 141 mg, 0.74 mmol) and Nl , N2-dimethylethane- 1 ,2-diamine (79 L, 0.73 mmol) and the reaction was then heated at 100°C for 2 days and then cooled to room temperature. Ethyl acetate was added; the reaction was filtered through celite and the filtrate was reduced in vacuo. Purification by silica gel chromatography provided intermediate 8-b as an off-white solid. MS (m/z) M+H= 470.4

Step 3: Compound 11 To a solution of intermediate 8-b ( 150 mg, 0.32 mmol) in ethanol (4.0 mL) and DMSO ( 1 .0 mL) were sequentially added NaOH I N (2.56 mL, 2.56 mmol) and 30% aqueous hydrogen peroxide ( 131 L, 1 .27 mmol) and the reaction was stirred at 50°C overnight and then cooled to room temperature. Water was added, a precipitate formed and compound 1 1 was isolated by filtration as a white solid. MS (m/z) M+H= 488.5

Synthesis of compound 13

Scheme 9

Step 1: 9-a

To a solution of N-Boc-glycine ( 130 mg, 0.74 mmol) in CH2CI2 cooled to 0°C were sequentially added EDC ( 1 0 mg, 1.0 mmol), and DMAP (catalytic). After stirring for 10 minutes compound 9 ( 150 mg, 0.33 mmol) was added and the reaction mixture was then stirred overnight at room temperature. Water and ethyl acetate were added; the organic layer was separated, washed with brine, dried over anhydrous MgSC>4, filtered and concentrated in vacuo. Purification by silica gel chromatography provided intermediate 9-a as a white solid. MS (m/z) M+H= 61 1 .5

Step 2: Compound 13

To a solution of intermediate 9-a (200 mg, 0.32 mmol) in CH2CI2 (2 raL) was added 2,2,2-trifluoroacetic acid ( 1 .5 raL) and the reaction was then stirred at room temperature for 2 hours. Volatiles were removed under reduced pressure and the residue was triturated with diethyl ether to provide compound 13.TFA as a white solid. MS (m/z) M+H= 51 1 .7

Synthesis of compound 12

Scheme 10

Compound 12

To a solution of compound 9 ( 100 mg, 0.22 mmol) in THF were sequentially added DIPEA ( 193 uL, 1 .10 mmol) and 1 , 1 '-carbonyldiimidazole ( 107 mg, 0.66 mmol) and the reaction was stirred at 50°C overnight. N,N-dimethylethylenediamine (97 uL, 0.88 mmol) was then added, the reaction was refluxed overnight and then cooled to room temperature. Volatiles were removed in vacuo. Purification by silica gel chromatography provided compound 12 as a yellow solid. MS (m/z) M+H= 568.5 Synthesis of compound 27

Scheme 11

Step 1: 11-b

To a solution of ( l r,4r)-4-aminocyclohexanecarboxylic acid (2.0 g, 13.97 mmol) 2N NaOH ( 14 mL) was added dropwise benzyl chloroformate (2.12 mL, 14.88 mmol) and the reaction was then stirred at room temperature for 1 .5 hours. The mixture was acidified to pH=3 with 1.0 N HC1 and then diluted with water. Intermediate 1 1 -b was collected by filtration as a white solid.

Step 2: 11-c

To a solution of intermediate 1 1 -b (2.0 g, 7.21 mmol) in DMF were sequentially added tBuOH (2.76 mL, 28.8 mmol), DMAP (352 mg, 2.88 mmol) and 1 ,3- diisopropylcarbodiimide (2.48 mL, 15.87 mmol) and the reaction was stirred at room temperature for 4 days. Water and ethyl acetate were added; the organic layer was separated, washed with 10% citric acid, saturated aqueous NaHCO^ and brine, dried over anhydrous MgS0 ₄ , filtered and concentrated in vacuo. Purification by silica gel chromatography provided intermediate 1 1 -c as a white solid.

Step 3: 11-d

To a solution of intermediate 1 1 -c ( 1 .2 g, 3.60 mmol) in MeOH and stirred under N2 was added 10 % Pd/C (38 mg). The reaction mixture was purged with ¾ and stirred for 2 hours. The reaction was then filtered through ceiite and the filtrate was concentrated in vacuo to provide intermediate 1 1 -d as a white solid. MS (m/z) M+H= 200.0

Step 4: 11-e

To a suspension of intermediate 1 1 -d (357 mg, 1.79 mmol) and 4-bromo-2,6- diflurobenzonotrile 5-d (391 mg, 1.79 mmol) in DMSO was added triethylamine (553 uL, 3.94 mmol) and the reaction was then stirred at room temperature for 2 days. Saturated aqueous ammonium chloride and ethyl acetate were added; the organic layer was separated, dried over anhydrous MgSC>4, filtered and concentrated in vacuo. Purification by silica gel chromatography provided intermediate 1 1 -e as a white solid.

Step 5: 11-f

To a solution of intermediate 1 1 -e (2.20 g, 5.54 mmol) and intermediate 5-c ( 1 .12 g, 5.54 mmol) in dioxane were sequentially added potassium carbonate (3.83 g, 27.7 mmol), copper (I) iodide ( 1 .58 g, 8.31 mmol) and Nl , N2-dimethylethane- l ,2-diamine (596 L, 5.54 mmol) and the reaction was then heated at 1 10°C for 2 days and then cooled to room temperature. Ethyl acetate was added; the reaction was filtered through celite and the filtrate was reduced in vacuo. MeOH was added to the residue and intermediate 1 1 -f was isolated by filtration as yellow solid. MS (m/z) M+H= 520.6

Step 6: 11-g

To a solution of intermediate 1 1 -f ( 180 mg, 0.34 mmol) in methanol ( 10.0 mL) and DMSO ( 10 mL) were sequentially added NaOH I N (346 L, 0.34 mmol) and 30% aqueous hydrogen peroxide (53 L, 0.52 mmol) and the reaction was stirred at room temperature for 1 hour. Saturated aqueous ammonium chloride and ethyl acetate were added, the organic layer was separated, washed with brine, dried over anhydrous MgSC>4, filtered and concentrated in vacuo to provide intermediate 1 1 -g as a white solid. MS (m/z) M+H= 538.5

Step 7: 11-h

To a solution of intermediate 1 1 -g ( 180 mg, 0.33 mmol) in dichloromethane (5 mL) was added 2,2,2-trifluoroacetic acid (5 mL) and the reaction was then stirred at room temperature for 1 hour. Volatiles were removed under reduced pressure and the residue was triturated with diethyl ether to provide intermediate 1 1 -h as a white solid. MS (m/z) M+H= 482.5

Step 9: Compound 27

To a solution of intermediate 1 1 -h (40 mg, 0.08 mmol) in DMF were sequentially added HATU (32 mg, 0.08 mmol), 2-morpholinoethanamine (22 mg, 0.16 mmol), HO AT ( 14 L, 0.08 mmol) and triethylamine (46 L, 0.33 mmol) and the reaction was then stirred for 30 minutes at room temperature. Saturated aqueous ammonium chloride and ethyl acetate were added; the organic layer was separated, washed with saturated

NaHCC and brine, dried over anhydrous MgSC>4, filtered and concentrated in vacuo. Purification by silica gel chromatography provided compound 27 as a white solid. MS (m/z) M+H=594.5 Synthesis of compound 20

Scheme 12

Step 1: 12-a

A solution of 4-bromo-2-fluorobenzonitrile 7-d (2.91 g, 14.56 mmol), tert-butyl 4- aminocyclohexylcarbamate (3.12 g, 14.56 mmol) and triethylamine (6.13 mL, 43.70 mmol) in DMSO was heated at 130°C for 5 hours and then cooled to room temperature. Saturated aqueous ammonium chloride and ethyl acetate were added; the organic layer was separated, washed with saturated aqueous ammonium chloride and brine, dried over anhydrous MgS0 ₄ , filtered and concentrated in vacuo. Diethyl ether was added to the residue and intermediate 12-a was collected by filtration as a white solid.

Step 2: 12-b

To a solution of intermediate 12-a (4.34 g, 9.84 mmol) and intermediate 5-c (2.0 g, 9.84 mmol) in 1 ,4-dioxane were sequentially added potassium carbonate (6.80 g, 49.2 mmol), copper (I) iodide (2.81 g, 14.76 mmol) and Nl , N2-dimethylethane- 1 ,2-diamine ( 1.05 raL, 9.84 mmol) and the reaction was then heated at 1 10°C for 2 days and then cooled to room temperature. Ethyl acetate was added; the reaction was filtered through celite and the filtrate was reduced in vacuo. Diethyl ether was added to the residue and intermediate 12-b was isolated by filtration as yellow solid. MS (m/z) M+H= 517.6

Step 3: 12-c

To a solution of intermediate 12-b (2.44 g, 4.72 mmol) in ethanol (47.2 mL) and DMSO ( 1 1.8 mL) were sequentially added NaOH IN (9.45 mL, 9.45 mmol) and 30% aqueous hydrogen peroxide ( 1.44 mL, 14.17 mmol) and the reaction was stirred at 50°C for 1 hour and then cooled to room temperature. Water was added and intermediate 12-c was isolated by filtration as a white solid. MS (m/z) M+H= 535.6

Step 4: 12-d

To a solution of intermediate 12-c (2.0 g, 3.74 mmol) in DCM ( 1 1.5 mL) was added 2,2,2-trifluoroacetic acid ( 1 1.5 mL) and the reaction was then stirred at room temperature for 1 hour. Volatiles were removed under reduced pressure and the residue was triturated with diethyl ether to provide intermediate 12-d TFA as a yellow solid. MS (m/z) M+H= 435.6

Step 5: Compound 20

To a solution of intermediate 12-d ( 100 mg, 0.23 mmol) in THF were sequentiaaly added DIPEA ( 121 L, 0.69 mmol) and l , l '-carbonyk1iimidazole (75 mg, 0.46 mmol) and the mixture was stirred at 50°C overnight. 2-(Pyrrolidin- l -yl)ethanamine ( 146 L, 1 .15 mmol) was added and the reaction was stirred at 50°C for an additional 3 hours and then cooled to room temperature. Volatiles were removed under reduced pressure.

Purification by silica gel chromatography provided compound 20 as a white solid. MS (m/z) M+H= 575.5 Synthesis of compound 30

To a solution of intermediate 12-d TFA ( 100 mg, 0.18 mmol) in DCM were sequentially added 3-(dimethylamino)propanoic acid (32 mg, 0.27 mmol), EDC (52 mg, 0.27 mmol), DMAP (2 mg) and triethylamine (51 L, 0.36 mmol) and the reaction was then stirred at room temperature overnight. Saturated aqueous ammonium chloride and ethyl acetate were added; the organic layer was separated, washed with saturated aqueous NaHCC and brine, dried over anhydrous MgSC>4, filtered and concentrated in vacuo. Purification by silica gel chromatography provided compound 30 as a white solid. MS (m/z) M+H= 534.6

Synthesis of compound 40

Scheme 14

Step 1: 14-b

A solution of intermediate 4-bromo-2,6-difluorobenzonitrile 1 1 -a (5.04 g, 23.14 mmol), (s)-3-aminotetrahydrofuran tosylate (6.0 g, 23.14 mmol) and DIPEA ( 12.12 raL, 69.4 mmol) in DMSO was stirred at room temperature for 2 days. Saturated aqueous ammonium chloride and ethyl acetate were added; the organic layer was separated, washed with saturated aqueous ammonium chloride and brine, dried over anhydrous MgS0 ₄ , filtered and concentrated in vacuo. Hexane was added and intermediate 14-b was collected by filtration as yellow solid.

Step 2: 14-c

To a solution of intermediate 14-b ( 1 .33 g, 4.69 mmol) and intermediate 6-c ( 1 .0 g, 4.92 mmol) in toluene were sequentially added potassium phosphate (2.08 g, 9.84 mmol), copper (I) iodide ( 178 mg, 0.93 mmol) and Nl , N2-dimethylethane- 1 ,2-diamine ( 195 L, 1.96 mmol), the reaction was then heated at 1 10°C overnight and then cooled to room temperature. Saturated aqueous ammonium chloride and ethyl acetate were added; the organic layer was separated, washed with saturated aqueous ammonium chloride and brine, dried over anhydrous MgSC>4, filtered and concentrated in vacuo. Purification by silica gel chromatography provided intermediate 14-c as a yellow solid. MS (m/z) M+H= 408.4 Step 3: Compound 40

To a solution of intermediate 14-c (450 mg, 1 .10 mmol) in MeOH ( 10.6 raL) and DMSO ( 10.6 raL) were sequentially added NaOH IN ( 1.10 raL, 1.10 mmol) and 30% aqueous hydrogen peroxide ( 169 L, 1 .65 mmol), the reaction was stirred at room temperature for 1 hour. Water and ethyl acetate were added; the organic layer was separated, washed with saturated aqueous ammonium chloride and brine, dried over anhydrous MgS0 ₄ , filtered and concentrated in vacuo. Purification by silica gel chromatography provided compound 40 as a white solid. MS (m/z) M+H= 426.5

Step 1 : 15-a

To a solution of intermediate 14-b ( 1 .65 g, 5.80 mmol) and intermediate 5-c ( 1 .25 g, 6.09 mmol) in toluene were sequentially added potassium phosphate (2.59 g, 12.18 mmol), copper (I) iodide (221 mg, 1.16 mmol) and Nl , N2-dimethylethane- l ,2-diamine (499 L, 4.64 mmol), the reaction was then heated at 1 10°C for 2 days and then cooled to room temperature. Saturated aqueous ammonium chloride and ethyl acetate were added; the organic layer was separated, washed with saturated aqueous ammonium chloride and brine, dried over anhydrous MgSC>4, filtered and concentrated in vacuo. Purification by silica gel chromatography provided intermediate 15-a as a yellow solid. MS (m/z) M+H= 410.4 Step 2: Compound 40

To a solution of intermediate 15-a (560 mg, 1 .36 mmol) in MeOH ( 13.1 mL) and DMSO ( 13.1 mL) were sequentially added NaOH IN ( 1.36 mL, 1.36 mmol) and 30% aqueous hydrogen peroxide (210 L, 2.05 mmol), the reaction was stirred at room temperature for 1 hour. Water and ethyl acetate were added; the organic layer was separated, washed with saturated aqueous ammonium chloride and brine, dried over anhydrous MgS0 ₄ , filtered and concentrated in vacuo. Purification by silica gel chromatography provided compound 40 as a white solid. MS (m/z) M+H= 428.5

Other compounds of the instant invention include the following:

Cancer Cell Survival Assay - HCT116 cell

Colorectal carcinoma HCT1 16 cells (ATCC# CCL-247) were cultured as monolayers in 96 well plates at a density of 2000 cells per well in McCoy's 5a medium (HyClone) supplemented with 2.2 g/L sodium bicarbonate (Gibco), 10% FBS (HyClone) and 1 % penecillin/streptomycin (HyClone). After 24 hour incubation, triplicate wells were treated with various concentrations of compound. Cells were incubated in the presence of compound for 72 hours at 37 °C, 5% CO ₂ . Metabolic viability of remaining cells was assessed by MTT (thiazolyl blue tetrazolium bromide, Sigma) assay.

Determination of EC50 Values from Cell Survival Curves

EC50 values (50% cell survival in the presence of compound as compared to untreated controls) were calculated from survival curves using BioAssay software (CambridgeSoft). Fluorescence polarization HSP90 binding assay

Fluorescence polarization based HSP90 binding assay was performed using modifications to previously described methods using full length HSP90 and a

geldanamycin-FITC probe (see Llauger-Bufi, L. et al., Bioorg. Med. Chem. Lett. 13

(2003) 3975-3978). Briefly, geldanamycin-FITC probe was diluted into HFB buffer [20 mM HEPES (K) pH 7.3, 50 mM KC1, 1 mM DTT, 5 mM MgCl ₂ , 20 mM Na ₂ Mo0 ₄ , 0.01 % NP40, 0.1 mg/mL of Bovine gamma-globuline] to obtain a working concentration of 8 nM.. HSP90 protein (Hsp9() Native Protein, Stressgen, SPP-770) was diluted in order to obtain a 4X stock protein solution into HFB buffer. The final amount of protein used in the assay corresponds to the amount of protein necessary to obtain 80% of the maximum FP value in a 2 nM probe saturation experiment. Assay were carried out in duplicates, into not treated black 96-well plate (Corning #3915), in a total volume of 100 μΐ, for a final concentration of 2 nM of geldanamycin-FITC probe, various concentrations of compound and Hsp-90 protein into HFB buffer. Buffer only (blank) or probe only in buffer (G-factor) were also added to be used as controls for calibration. The plate was left on a shaker at 4 °C for 3 hours and the FP values in mP were recorded using Genios Pro FP reader (TECAN). The measured FP values (mP) were then plotted against compound concentration and EC ₅₀ , corresponding to the competitor concentrations where 50% of the tracer was displaced, calculated based on a sigomoidal dose-response (variable-slope) curve fit using GraphPad Prism version 4.02 for Windows, GraphPad Software, San Diego California USA, www.graphpad.com.

Several representative compounds of the invention are listed below and their respective magnitude of potency is provided.

HCT1 16 EC ₅ o and HSP90 binding IC50 A: less than 100 nM; B between 100 and

1000 nM; C greater than 1000 nM.

Western Blot Analysis of HSP70 Levels

RAW-Blue mouse macrophage cells (InvivoGen, grown in DMEM, 10% heat inactivated FBS, 1 % penecillin/streptomycin, HyClone) were seeded in 6 well plates one day prior to treatment. Cells were treated with various concentrations of compound for 18 hours at 37° C, 5% CO ₂ . Lysates were prepared in RIPA buffer. Equal amounts of clarified extracts were resolved on 10% SDS-PAGE gels, electrophoretically transferred to PVDF membranes and blocked with 5% milk prior to incubation with the following antibodies: anti-HSP70 (W27 Clone AB-2, Neomarkers, 1/2000) or GAPDH (clone 6C5, Advanced Immunochemical, 1/25,000) antibodies. Immunoreactive signals were detected by incubation with horseradish peroxidase-conjugated secondary antibody (Chemicon) followed by chemiluminescent detection of immunoreactive proteins.

Compound 4 induced HSP70 in a dose dependent manner at concentration of less than 10 μΜ. EC50 determination for inhibition of LPS triggered TNF-alpha release in human whole blood in- vitro

Human whole blood was collected in Vacutainer Lithium Heparin blood collection tubes (BD Biosciences) from healthy donors. Various concentrations of compound (25 uL) diluted in RPMI media containing 10% FBS was added to 5()() L aliquot of whole blood in 2 mL micro-tubes (Sarstedt) and incubated in rotation at 37 °C for 4 hours. LPS (25 uL, 2 ug/mL, Sigma) was added to blood aliquots and incubated for 2 hours in the same conditions. At the end of incubation, plasma was prepared from blood aliquots by centrifugation in microfuge at 5()()g for 10 minutes. TNF-alpha content of plasma samples was measured using commercial TNF-alpha ELISA kit (BD Biosciences).

Compounds 16 and 37 demonstrated EC ₅₀ of less than 10 uM demonstrating that compounds of formula 1 are capable of reducing an inflammatory response in human whole blood.

EC50 determination for inhibition of LPS triggered TNF-alpha release in human PBMC in- vitro

Peripheral blood mononuclear cells (PBMCs) were isolated from Lithium Heparin blood from healthy donor with a standard density gradient centrifugation (Histopack 1 .077; Sigma-Aldrich). The isolated PBMCs were washed two times in phosphate buffered saline (PBS; pH 7.4). PBMC were resuspended in 37 °C RPMI media supplemented with 10% FBS and 0.1 % penicillin/streptavidin. Viable PBMC cells were counted manually with a hemacytometer in presence of trypan blue and diluted to 1() ⁶ cells per ml in supplemented RPMI media. PBMC are seeded at όχ ΐ θ cells per well in 48-well cell culture plate. Various concentrations of compound diluted in RPMI media containing 10% FBS (30 uL) were added to PBMC and incubated 18 hours at 37 °C in humidified atmosphere in cell incubator. LPS (25 uL, 2 ug/mL, Sigma) prepared in RPMI media supplemented with 10% FBS and 0.1 % P/S was added to PBMC and incubated at 37 °C for two hours. At the end of the incubation, 250 uL of media from each sample well were collected and TNF-alpha content was measured using commercial TNF-alpha ELISA kit (BD Biosciences). Compounds 16 and 37 demonstrated EC ₅₀ of less than 10 uM demonstrating that compounds of formula 1 are capable of reducing an inflammatory response in human PBMCs.

Induction of HSP70 in Peripheral Blood Mononuclear Cells (PBMCs)

Female Swiss mice (5 per group) received 5 consecutive daily PO doses ( 10, 30, 60 or 100 mg/kg) of compound. Trunk blood was collected in vacutainer containing EDTA anticoagulant 24h after the final dose. Blood from mice of the same group was pooled and peripheral blood mononuclear cells (PBMC) were isolated with a density separation medium (Lympholyte-Mammal, Cedarlane) and lysed in RIPA buffer. HSP70 protein levels were measured using a commercial sandwich ELISA (R&D Systems).

Compound 16, administered as compound 37, was shown to induce a dose dependent increase in HSP70 protein in PBMCs (up to a 15 fold) demonstrating that compounds of formula 1 are capable of inducing HSP70 in vivo.

Induction of HSP70 in brain

Female Swiss mice (3 per group) received 5 consecutive daily PO doses of compound (60 or 100 mg/kg). Brains were collected 6 hours after the final dose, homogenized using a motorized pellet pestle and lysed in RIPA buffer. HSP70 protein levels were measured using a commercial sandwich ELISA (R&D Systems).

Compound 16, administered as compound 37, was shown to induce a dose dependent increase in HSP70 protein in brain homogenate (up to a 2 fold) demonstrating that compounds of formula 1 are capable of inducing HSP70 in the brain.

Pharmacokinetics (PK) of Compound in Plasma and Brain

The concentrations of compounds of formula 1 , such as compound 16, administered as compound 37, were determined in the plasma and brain homogenate of mice after treating CD- I mice with compound via IV or oral administration. Compound concentrations were determined using acetonitrile extraction of compound from the appropriate matrix and calculation of drug concentrations using a spiked standard curve from the same matrix (ie. plasma or brain homogenate).

Compounds of formula 1 demonstrated good oral absorption and exposure in brain. These results indicate that compounds of formula 1 are orally available and can cross the blood brain barrier. Therefore compounds of formula 1 may be useful in the treatment of both peripheral and CNS related diseases and conditions.

Equivalents

Those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, numerous equivalents to the compounds and methods of use thereof described herein. Such equivalents are considered to be within the scope of this invention and are covered by the following claims.

All of the above-cited references and publications are hereby incorporated by reference.