WO2008115259A2

WO2008115259A2 - Derivatives of benzoxadiazole suitable for the treatment of cell proliferative diseases

Info

Publication number: WO2008115259A2
Application number: PCT/US2007/075511
Authority: WO
Inventors: Rajinder Singh; Usha Ramesh; Jianing Huang; Sarkiz D. Issakani; Lyuben Tsvetkov; Matthew David Petroski
Original assignee: Rigel Pharmaceuticals Inc
Current assignee: Rigel Pharmaceuticals Inc
Priority date: 2006-08-10
Filing date: 2007-08-08
Publication date: 2008-09-25
Anticipated expiration: 2009-02-10
Also published as: WO2008115259A3

Abstract

This invention describes compounds and pharmaceutical compositions useful as ubiquitin agent inhibitors, particularly ubiquitin ligase inhibitors. The compounds and pharmaceutical compositions of the invention are useful as inhibitors of the biochemical pathways of organisms in which ubiquitination is involved, such as signal transduction pathways. The invention also comprises the use of the compounds and pharmaceutical compositions of the invention for the treatment of conditions that require inhibition of ubiquitination. Furthermore, the invention comprises methods of inhibiting ubiquitination in a cell comprising contacting a cell in which inhibition of ubiquitination is desired with a compound or pharmaceutical composition according to the invention. Particularly, the compounds and pharmaceutical compositions are useful to inhibit the ubiquitin ligase activity of TRAF6.

Description

UBIQUITIN LIGASE INHIBITORS

BACKGROUND OF THE INVENTION Field of the Invention

[0001] This invention relates to the inhibition of ubiquitination. More particularly, the invention relates to compounds and methods for inhibiting ubiquitin ligase activity.

Summary of the Related Art

[0002] Ubiquitin is a 76 amino acid protein present throughout the eukaryotic kingdom. It is a highly conserved protein and is essentially the identical protein in diverse organisms ranging from humans to yeasts to fruit flies. In eukaryotes, ubiquitination is the key component of the ATP-dependent pathway for protein degradation and cellular regulatory processes. Proteins slated for degradation or that act as regulatory agents are covalently linked to ubiquitin via an ATP-dependent process catalyzed by three separate enzymes.

[0003] The ubiquitination of these target proteins is known to be mediated by the enzymatic activity of three ubiquitin agents. Ubiquitin is first activated in an ATP-dependent manner by a ubiquitin activating agent, for example, an El. The C-terminus of a ubiquitin forms a high energy thioester bond with the ubiquitin activating agent. The ubiquitin is then transferred to a ubiquitin conjugating agent, for example, an E2 (also called ubiquitin moiety carrier protein), also linked to this second ubiquitin agent via a thioester bond. The ubiquitin is finally linked to its target protein (e.g. substrate) to form a terminal isopeptide bond under the guidance of a ubiquitin ligating agent, for example, an E3. In this process, monomers or oligomers of ubiquitin are attached to the target protein. On the target protein, each ubiquitin is covalently ligated to the next ubiquitin through the activity of a ubiquitin ligating agent to form polymers of ubiquitin. [0004] Typically, the ubiquitination of target proteins by E3 in cells results in the formation of poly-ubiquitin chains. An isopeptide bond is formed between the carboxyl terminus of the ubiquitin and the ε-amino group of Lys in the target protein. The extension or formation of ubiquitin chains results from the formation of additional isopeptide bonds with the Lys⁴⁸ (and sometimes Lys⁶³) of a previously conjugated ubiquitin and the carboxyl-terminal GIy of an additional ubiquitin. The efficient recognition of a ubiquitinated target protein by a proteosome requires at least four ubiquitins linked in this configuration. However, in the case of Mdm2 -mediated ubiquitination of p53, neither Lys⁴⁸ or Lys⁶³ is involved in the formation of poly-ubiquitin chains. Recent studies show that human Mdm2 mediates multiple mono-ubiquitination of p53 by a mechanism requiring enzyme isomerization (Zhihong et al. (2001) J.Biol.Chem. 276:31,357-31,367). Further, in vitro, the transfer of ubiquitin to p53 can occur independent of El when using an E2 pre-conjugated with ubiquitin. These results suggest that the pre-conjugated E2 can bind to Mdm2 and thereafter transfer the ubiquitin to the Mdm2 in the absence of an E 1.

[0005] The enzymatic components of the ubiquitination pathway have received considerable attention (for a review, see Wong et al, DDT 8:746-754 (2003); Weissman, Nature Reviews 2:169-178 (2001)). The members of the El ubiquitin activating agents and E2 ubiquitin conjugating agents are structurally related and well characterized enzymes. There are numerous species of E2 ubiquitin conjugating agents, some of which act in preferred pairs with specific E3 ubiquitin ligating agents to confer specificity for different target proteins. [0006] The family of ubiquitin and ubiquitin-like modifiers includes ubiquitin, NEDD8, ISG 15, SUMOl, SUM02, SUM03, APG 12, and APG8. Further, genome mining efforts have identified at least 530 human genes that encode enzymes responsible for ubiquitin conjugation and deconjugation. Many of these genes encode multiple splice variants, thereby increasing the diversity of enzyme families regulating ubiquitin conjugation and deconjugation. There are a multitude of E3's, reflecting their role as specificity determinants, an intermediate number of E2's, and few El's, which are redundant to multiple pathways. Thus, the same E2 in conjunction with different E3's recognizes distinct substrates. For example, ubiquitin and ubiquitin-like enzymes are encodes by at least 11 genes that comprise at least 11 isoforms; El 's are encoded by at least 13 genes that include at least 15 isoforms; E2's, which include Ubc (ubiquitin carrier proteins) and Uev (ubiquitin enzyme variants), are encoded by at least 49 genes that comprise at least 77 isoforms; E3's, which include RING, PHD, HECT and U-box domain containing proteins, are encoded by at least 391 genes that comprise at least 631 isoforms; and DUB 's (de-ubiquitylating enzymes), which includes USP, ULP, JAMM and UCH proteases, are encoded by at least 86 genes that comprise at least 136 isoforms. Ubiquitin conjugation and deconjugation pathways regulate diverse biological pathways. For example, ubiquitin conjugation and deconjugation pathways play important roles in cancers, inflammation, metabolism, viral diseases and central nervous system disorders. Thus, compounds that can modulate ubiquitin conjugation and deconjugation processes would serve as important therapeutic agents. For a more detailed review of ubiquitin regulatory pathways see Wong et al. (DDT 8 (16), 746-754 (2003)).

[0007] Generally, ubiquitin ligating agents contain two separate activities: a ubiquitin ligase activity to attach, via an isopeptide bond, monomers or oligomers of ubiquitin to a target protein, and a targeting activity to physically bring the ligase and substrate together. The substrate specificity of different ubiquitin ligating agents is a major determinant in the selectivity of the ubiquitin-mediated protein degradation process.

[0008] In eukaryotes, some ubiquitin ligating agents contain multiple subunits that form a complex having ubiquitin ligating activity. Particularly well characterized examples are SCFs, which play an important role in regulating Gl progression and consists of at least three subunits: SKPl, Cullins (having at least seven family members) and an F-box protein (of which hundreds of species are known) which bind directly to and recruit the substrate to the complex. The combinatorial interactions between the SCF 's and a recently discovered family of RING finger proteins, the ROC/ APC 11 proteins, have been shown to be the key elements conferring ligase activity to ubiquitin ligating agents. Particular ROC/Cullin combinations can regulate specific cellular pathways, as exemplified by the function of APCl 1-APC2, involved in the proteolytic control of sister chromatid separation and exit from telophase into Gl in mitosis {see King et al, supra; Koepp et al, Cell 97:431-34 (1999)), and ROCl-Cullin 1, involved in the proteolytic degradation of 1KB in NF-KB/IKB mediated transcription regulation (Tan et al, Mol Cell 3(4):527-533 (1999); Laney et al, Cell 97:427-30 (1999)).

[0009] The best characterized ubiquitin ligating agent is the APC (anaphase promoting complex), which is multi-component complex that is required both for entry into anaphase as well as exit from mitosis {see King et al, Science 274:1652-59 (1996) for review). The APC plays a crucial role in regulating the passage of cells through anaphase by promoting ubiquitin-mediated proteolysis of many proteins. In addition to degrading the mitotic B-type cyclin for inactivation of CDC2 kinase activity, the APC is also required for degradation of other proteins for sister chromatid separation and spindle disassembly. Most proteins known to be degraded by the APC contain a conserved nine amino acid motif known as the "destruction box" that targets them for ubiquitin ubiquitination and subsequent degradation. However, proteins that are degraded during Gl, including Gl cyclins, CDK inhibitors, transcription factors and signaling intermediates, do not contain this conserved amino acid motif. Instead, substrate phosphorylation appears to play an important role in targeting their interaction with a ubiquitin ligating agent for ubiquitination (see Hershko et ah, Ann. Rev. Biochem. 67:429-75 (1998)). [0010] Two major classes of E3 ubiquitin ligating agents are known: the HECT (homologous to E6-AP carboxy terminus) domain E3 ligating agents; and the RING finger domain E3 ligating agents. E6AP is the prototype for the HECT domain subclass of E3 ligating agents and is a multi-subunit complex that functions as a ubiquitin ligating agent for the tumor suppressor p53 which is activated by papillomavirus in cervical cancer (Huang et al. (1999) Science 286:1321-1326). Members of this class are homologous to the carboxyl terminus of E6AP and utilize a Cys active site to form a thiolester bond with ubiquitin, analogous to the El activating agents and E2 conjugating agents. However, in contrast, the members of the RING finger domain class of E3 ligating agents are thought to interact with an ubiquitin-conjugated-E2 intermediate to activate the complex for the transfer of ubiquitin to an acceptor. Examples of the RING domain class of E3 ligating agents are TRAF6, involved in IKK activation; CbI, which targets insulin and EGF; Sina/Siah, which targets DCC; Itchy, which is involved in haematopoesis (B, T and mast cells); IAP, involved with inhibitors of apoptosis; and Mdm2 which is involved in the regulation of p53.

[0011] The RING finger domain subclass of E3 ligating agents can be further grouped into two subclasses. In one subclass, the RING finger domain and the substrate recognition domain are contained on different subunits of a complex forming the ubiquitin ligating agent {e.g., the RBxI and the F-box subunit of the SCF complex). In the second subclass of ubiquitin ligating agents, the ligating agents have the RING finger domain and substrate recognition domain on a single subunit. (e.g., Mdm2 and cbl) (Tyers et al. (1999) Science 284:601, 603-604; Joazeiro et al. (2000) 102:549-552). A further class of ligating agents are those having a "PHD" domain and are homo logs of the RING finger domain ligating agents (Coscoy et al. (2001) J. Cell Biol. 155(7): 1265-1273), e.g., MEKKl. The PHD domain ligating agents are a novel class of membrane-bound E3 ligating agents.

[0012] In addition, a new class of ubiquitin ligases has been characterized. These are the U-box-containing proteins. (Patterson, Sci STKE 2002(116:PE4 (220)). This class, for the present, represents a small number of ligases which have yet to be extensively characterized. [0013] Recently, an E2-E3 pair has been identified to consist of the UBC13-TRAF6 in conjuction with the Ubc-like protein Uevl A. This newly identified E2-E3 pair participates in many signal transduction pathways. TRAF6 (tumor necrosis factor (TNF) receptor associated factors) was first identified as intracellular proteins associated with TNF -R2 (reviewed by Wu et al, BioEssays 25, 1096-1105). Deng et al. (Cell 103, 351-361 (2000)) demonstrated that the RING domain of TRAF6, in conjunction with the ubiquitin-conjugating enzyme UBC 13 and the UBC-like protein Uevl A, exhibited ubiquitin ligase activity and catalyzed polyubiquitin chain formation linked by Lys-63, instead of Lys-48 of ubiquitin. It was later shown that the ubiquitin ligase activity of TRAF 6 promotes poly-ubiquitin chains formation that activates TAKl which in turn activates a number of important kinases such as IkB kinase and MAP kinases (reviewed by Wu et al., BioEssays 25, 1096-1105).

[0014] The ubiquitination of TRAF6 leads to activation of TAKl which then activates IkB kinase. IkB kinase in turn activates the NF-kB pathway as well as phosphorylates MKK6 in the JNK-p38 kinase pathway. The NF-kB pathway includes many important processes such as inflammation, LPS-induces septic shock, viral infection such as HIV, and cell survival among others. Thus, the ubiquitin ligase activity of TRAF 6 plays important regulatory roles in many cellular processes. Stem cells are characterized by their ability to self-renew as well as generate differentiated cells within each organ. There is increasing evidence that these cells or their immediate progeny may be targets for transformation and it has been hypothesized that an early event in carcinogenesis may involve dysregulation of stem cell self-renewal leading to a clonal expansion of initiated stem cells (see Dontu, et al. Cell Prolif 36:59-72 (2003); and Wicha et al., Cancer Res, 66:1883-90 (2006)). A number of developmental signaling pathways, such as Wnt, Notch, and hedgehog, have been found to play a role in regulating the self-renewal of normal stem cells in the hematopoietic system, the skin, the nervous system, and the breast (see Dontu, et al. Cell Prolif 36:59-72 (2003); Kopper and Hajdu Pathol Oncol ReslO:69-73 (2004); and Taipale and Beachy, Nature 411 :349-54 (2001)).

[0015] The "cancer stem cell hypothesis' ' postulates that tumors are driven by a cellular subpopulation retaining stem cell properties (see Wicha, et al. Cancer Res 66:1883-90, (2006); Kopper and Hajdu, Pathol Oncol Res, 10:69-73 (2004); and Reya, et al. Nature 414: 105-11, (2001)). Over the past several years, umorigenic stem cells have been detected in myeloma, brain cancers, sarcoma, and prostate cancers (see Pellat, et al. Blood Cells MoI Dis 32:293-301 (2004); Singh, et al. Cancer Res 63:5821-8 (2003); and Xin, et al. Proc Natl Acad Sci U S A 102:6942-7 (2005)), lending support to the cancer stem cell hypothesis. In vitro and mouse model systems have elucidated the role of hedgehog signaling and the polycomb gene Bmi-1 in regulating the self-renewal of normal human mammary stem cells. In normal mammary development, Hh and the downstream transcription factor Bmi-1 play an important role in regulating stem cell self renewal. These processes are tightly regulated by factors in the stem cell niche. Deregulation of these processes during carcinogenesis may result in stem cell expansion, a key event in carcinogenesis. The clinical importance of this is highlighted by a recent report demonstrating a strong correlation between the expression of an 11 -gene Bmi-1 stem cell signature and poor prognosis in patients with a wide variety of malignancies (see Glinsky, et al. J Clin Invest 115:1503-21 (2005)).

[0016] Bmi-1 was first isolated as an oncogene that cooperates with c-Myc in generating lymphomas in a murine model (see Haupt, et al. Cell 65:753-63 (1991); and van Lohuizen, et al. Cell 65:737-52 (1991)). It is a transcriptional repressor belonging to the Polycomb-group (PcG) family of proteins involved in axial patterning, hematopoiesis, regulation of proliferation, and senescence (see van der Lugt, et al. Genes Dev 8:757-69 (1994); and Pirrotta, Cell93:333-6 (1998)). Bmi-1 -deficient mouse embryonic fibroblasts (MEF) overexpress INK4a/ARF locus- encoded genes pl6INK4a and pi 9ARF (mouse homologue of human pl4ARF) and undergo premature senescence in culture (see Jacobs, et al. Nature 397:164-8 (1999)). Conversely, overexpression of Bmi-1 reduces expression of pl6 INK4a and pi 9ARF and immortalizes MEFs (see Jacobs, et al. Nature 397:164-8 (1999)). Recently, it has been found that Bmi-1 is overexpressed in a variety of human cancers, such as mantle cell lymphomas (see Bea, et al. Cancer Res 61 :2409-12 (2001)), non-small cell lung cancer (see Vonlanthen, et al. Br J Cancer 84:1372-6 (2001)), B-cell non-Hodgkin's lymphoma (see van Kemenade, et al. Blood 97:3896- 901, (2001)), breast cancer (see Kim, et al. Breast 13:383-8 (2004)), colorectal cancer (see Kim, et al. Cancer Lett 203:217-24 (2004)), and prostate cancer (see Glinsky, et al., J Clin Invest 115:1503-21 (2005)). It is has also been shown that Bmi-1 overexpression alone was able to immortalize post-selection pi 6INK4a-deficient human mammary epithelial cells (HMEC) and induced telomerase activity in these cells (see Dimri, et al. Cancer Res 62: 4736-45 (2002)). In human fibroblasts, which expresses pl6INK4a Bmi-1 overexpression results in extension of replicative life span but no immortalization (see Itahana, et al. MoI Cell Biol 23:389-401 (2003)). Bmi-1 was also reported to immortalize bone marrow stromal cells and cementoblast progenitor cells, albeit in combination with other oncogenes (see Saito, et al. J Bone Miner Res 20:50-7 (2005); Mori, et al. MoI Cell Biol 25:5183-905 (2005)).

[0017] Polycombgroup (PcG) proteins exist in at least twobiochemically distinct protein complexes, the EED-EZH2 complex and the PRCl complex, that respectively possess [0018] H3-K27 methyltransferase and H2A-K119 ubiquitin E3 ligase activities. Bmi-1 and Ring IA, two components of the PRCl complex, play important roles in H2A ubiquitylation and Hox gene silencing X-chromosome inactivation, tumorigenesis, and stem cell self-renewal; both proteins positively regulate H2A ubiquitylation. The RING finger protein Ring IB is an E3 ligase that participates in the ubiquitination of lysine 119 of histone H2A, and the binding of Bmi-1 stimulates the E3 ligase activity.

[0019] The PRCl complex has at least two biochemical functions. One of which is to bind chromatin and prevent it from being remodeled by ATP-dependent remodeling factors (see Shao, et al. Cell 98: 37-46, (1999)). Using an electron microscope, a Drosophila PRCl sub-complex has been seen to compact nucleosome arrays in vitro (see Francis, et al. Science 306: 1574-1577, (2004)). A recent study revealed that a human PRCl complex composed of Bmi-1, HPH2, PC3, and Ring proteins (RinglA and RinglB), which are homologs of Drosophila Posterior Sex Combs, Polyhomeotic, Polycomb, and dRing, respectively, is an E3 ubiquitin ligase complex that mono-ubiquitinates lysine 119 of nucleosomal histone H2A (seeWang, et al. Nature 431 : 873-878 (2004)). The catalytic subunit of the PRCl E3 ligase complex is RinglB. The E3 ligase activity has been shown to be important for the involvement of PRCl in X-chromosome inactivation and the control of Hox gene expression (see Fang, et al. J. Biol. Chem. 279: 52812- 52815 (2004); de Napoles, et al. Dev. Cell 7: 663-676 (2004); Hernandez-Munoz, et al. Proc. Natl. Acad. Sci. U. S. A. 102: 7635-7640 (2005); and Cao, et al. MoI. Cell 20: 845-854, (2005)). [0020] Chromatin immunoprecipitation (ChIP) assays demonstrate that Bmi-1 and other components of the two PcG complexes bind to the promoter of HoxC13. Knockout Bmi-1 results in significant loss of H2A ubiquitylation and upregulation of Hoxcl3 expression, whereas EZH2 -mediated H3-K27 methylation is not affected. Results suggest that EZH2- mediated H3- K27 methylation functions upstream of PRCl and establishes a critical role for Bmi-1 and RinglA in H2A ubiquitylation and Hox gene silencing.

[0021] Various types of DNA lesions generated by endogenous and environmental factors in living organisms can be repaired by various repair mechanisms (see Lindahl and Wood, Science, 286: 1897-1905, (1999)). However, if a lesion is not yet removed because of a limited repair capacity of cells and if the DNA replication machinery encounters this lesion before repair, then the replication machinery stalls at this lesion, causing a gap in the newly synthesized strand opposite to the damaged site. Further the gap could cause a secondary damage, much severer than the first one, such as DNA double-strand break (DSB), and thus could cause cell death unless the gap is filled. This gap filling is called as postreplication repair (PRR) and such gap filling function is conserved from bacteria through to human cells (see Friedberg, et al., Proc. Natl Acad. Sci. USA, 97: 5681-5683: (2000); and Lehmann Mutat. Res., 509: 23-34, (2002)). [0022] In the budding yeast Saccharomyces cerevisiae, genes belonging to the RAD6 epistasis group are involved in the PRR pathway (see Lawrence Bioessays, 16:253-258, (1994)). Among these genes, those encoding Rad6 (a ubiquitin-conjugating enzyme, E2) and Radl8 (a ubiquitin ligase, E3) play crucial roles in this pathway (see Bailly, et al. Genes Dev., 8: 811-820, (1994); and Bailly, et al. MoI. Cell. Biol, 17:4536-4543, (1997)). Yeast Radl8 was shown to bind to single-stranded DNA and also to form a tight complex with Rad6 (see Bailly, et al. Genes Dev., 8: 811-820, (1994); and Bailly,et al. J. Biol.Chem., 272: 23360-23365, (1997)) whereas Rad6 does not have any DNA-binding activity. Thus it was proposed that such Radl8 bound to the gap regions would recruit Rad6 to the replication-stalled sites, where Rad6 modulates the function of a stalled DNA replication machinery with its ubiquitin-conjugating activity (see Bailly et al. Genes Dev., 8: 811-820, (1994)).

[0023] Recently, in yeast cells, the proliferating cell nuclear antigen (PCNA), a DNA polymerase sliding clamp involved in DNA replication and repair, was shown to be monoubiquitinated at the lysine- 164 site in a Radl8/Rad6-dependent manner, which was necessary for carrying out translesion synthesis (TLS) by Rad30, a yeast homolog of polymerase η (poly η), which is a member of the RAD6 epistasis group (see Haracska, et al. MoI. Cell, 8: 407-415(2001); Hoege, et al. Nature, 419: 135-141, (2002); McDonald, et al. Genetics, 147: 1557-1568 (1997); and Stelter and Ulrich, Nature, 425, 188-191, (2003)). Similarly, monoubiquitination of human and mouse PCNAs was observed in a RAD18/RAD6-dependent manner at the sites of replication forks stalled by ultraviolet (UV) light-induced lesions, and caused a polymerase switch from a replicative to a translesion polymerase, pol η (see Kannouche, et al. MoI. Cell, 14: 491-500, (2004); and Watanabe, et al. EMBO J.,23: 3886-3896, (2004)). Thus, the mechanism of polymerase switching through the monoubiquitination of PCNA is conserved in various species.

[0024] Radl8 forms a tight complex with Rad6 both in yeast and vertebrate cells (see Tateishi, et al. Proc. Natl. Acad. Sci. U. S. A. 97: 7927-7932 (2000); and Bailly, V. et al, J. Biol. Chem. Ill: 23360-23365 (1997)). Rad6 is a member of the E2 and is also named UBC2. Yeast Rad6 is involved in amino end rule protein degradation through physical interaction with the E3 enzyme, Ubrl (see Dohmen, et al., Proc. Natl.

[0025] Acad. Sci. U. S. A. 88: 7351-7355 (1991)). Furthermore, Rad6 plays a role that regulates monoubiquitination of histone 2B with the help of Brel, an E3 protein containing one RING finger domain (see Joazeiro and Weissman, A. M. Cell 102: 549-552, (2000)). Like many other E3 enzymes containing the RING finger domain, both yeast and vertebrate Radl8 also contain it in the NH2 -terminal region, suggesting that Radl8 functions as an E3. Recently, it was found that Radl8 is involved in monoubiquitination of PCNA in cells treated with DNA- damaging agents and that PCNA monoubiquitination is critical for polymerase switching in translesion DNA synthesis (see Hoege, et al. Nature 419: 135-141(2002); Kannouche, et al., MoI. Cell 14 : 491-500(2004); and Stelter, and Ulrich Nature 425: 188-191(2003)). [0026] The mutation of either RAD 18 or RAD6 gene in yeast cells resulted in hypersensitivity to various DNA-damaging agents including UV light and methylmethanesulfonate (MMS) (see Hynes and Kunz, In Strathern, et al. (eds), The Molecular Biology of the Yeast Saccharomyces, Cold Spring Harbor Laboratory Press, Plainview, NY, pp. 371-414, (1981)), and caused the attenuation of the mutation-inducing ability following such exposure (see Broomfϊeld, et al. Mutat. Res., 486: 167-184, (2001)). In mammals, only a single homolog of yeast RAD18 has been identified (see Tateishi, et al. Proc. Natl Acad. Sci. USA, 97: 7927-7932, (2000)). The human and mouse RAD 18 proteins can interact with two forms of homo logs of yeast Rad6, RAD6A and RAD6B, both in vitro and in vivo (see Tateishi, et al. Proc. Natl Acad. Sci. USA, 97: 7927-7932, (2000); Tateishi, et al. MoI. Cell. Biol, 23: 474-481, (2003); and Xin, et al. Nucleic Acids Res., 28: 2847-2854, (2000)). Either overexpression of a dominant-negative RAD 18 mutant protein in human cells or targeted disruption of RAD 18 in mouse embryonic stem cells resulted in an increased sensitivity to various DNA damaging agents and also enhanced the genomic instability as determined by increases in sister-chromatic exchange (SCE) and also by stable transformation (see Tateishi, et al. Proc. Natl Acad. Sci. USA, 97: 7927-7932,(2000); and Tateishi, et al., MoL Cell. Biol, 23: 474-481 (2003)). [0027] Ubiquitination appears to switch PCNA activity from polymerization to repair mode. Inhibition of Radl8, preventing this switch, should make cancer cells more susceptible to radiation chemical DNA-damage cancer treatments.

[0028] Chu et al., have discussed the role of p27 phosphorylation of Src in regulating inhibition of cyclin E-Cdk2 (see Cell 128: 281-294 (2007)). Mammalian cell proliferation is tightly regulated by the sequential activation of cyclin-dependent kinases (Cdks) (see Sherr, et al. Genes Dev. 13: 1501-1512 (1999)). The Cdk inhibitor, p27, was initially identified in cells arrested by transforming growth_factor b (TGF -b), by contact inhibition, and by lovastatin (see Polyak et al., Genes Dev. 8 : 9-22 (1994); Hengst et al., Proc. Natl. Acad. Sci. USA 91 :5291- 5295 (1994); Slingerland et al., MoI. Cell. Biol. 14: 3683-3694. (1994); and Koff et al., Science 260: 536-539 (1993)). p27 is ubiquitously expressed and binds cyclin-Cdk2 to inhibit kinase activity.

[0029] p27 levels regulate cell proliferation and differentiation. p27 null animals exhibit multiorgan hyperplasia (see Fero et al., Nature 396: 177-180 (1996); Kiyokawa et al., Cell 85: 721-732 (1996); and Nakayama et al., Cell 85: 707-720 (1996)), and p27+/_ heterozygous animals are tumor prone (see Fero et al., Nature 396: 177-180 (1998)). While normal quiescent epithelial tissues express high p27 (see Catzavelos et al., Nat. Med. 3: 227-230 (1997)), reduced p27 protein is observed in up to 60% of primary human breast and other cancers in association with poor patient outcome (see Alkarain et al., J. Mammary Gland Biol. Neoplasia 9: 67-80 (2004)). Several oncogenic pathways activate p27 proteolysis in cancers (see Liang and Slingerland, Cell Cycle 2: 339-345 (2003)). Human breast cancers frequently show EGFR overexpression or Her2/ErbB2 amplification (see Tsutsui et al., Breast Cancer Res. Treat. 71: 67-75 (2002); Nicholson et al., J. Steroid Biochem. MoI. Biol. 37: 811-814 (1990); and Slamon et al., Science 235: 177-182 (1987)). Overexpression of EGFR or Her2 increases p27 proteolysis in cell lines (see Lane et al., MoI. Cell. Biol. 20: 3210-3223 (2000); Yang et al., J. Biol. Chem. 275: 24735-24739 (2000); and Lenferink et al., Proc. Natl. Acad. Sci. USA 97: 9609-9614 (2000)). Activated EGFR family receptor tyrosine kinases (RTK) recruit and activate cSrc, and cSrc in turn further activates RTKs, stimulating cell proliferation (see Ishizawar and Parsons, Cancer Cell 6: 209-214 (2004)). Drug-mediated cSrc inhibition blocks the effects of EGFR and Her2 on cell proliferation (see Belsches- Jablonski et al., Oncogene 20: 1465-1475 (2001); and Biscardi et al., J. Biol. Chem. 274:8335-8343 (1999)). cSrc is also activated by liganded estrogen receptor (ER) in human breast cancer cells. Estrogen:ER binding stimulates rapid, transient recruitment of cSrc, She activation, and MAPK signaling (see Migliaccio et al., EMBO J. 15: 1292-1300 (1996)). Estrogen:ER-stimulated Src further recruits RTKs, Her2, EGFR (see Chu et al., Cancer Res. 65: 18-25 (2005)), and IGF-IR (see Song et al., Proc. Natl. Acad. Sci. USA 101 : 2076-2081 (2004)), to promote cell cycle progression.

[0030] ER blockade by tamoxifen or estrogen deprivation by aromatase inhibitors is of therapeutic utility in a majority of breast cancers that express the ER. p27 is required for Gl arrest by tamoxifen or estrogen deprivation (see Cariou et al., Proc. Natl. Acad. Sci. USA 97: 9042-9046 (2000)). Despite initial responses, the development of antiestrogen resistance limits treatment efficacy. EGFRand Her2/ErbB2 overexpression can confer tamoxifen resistance in cultured lines (see Benz et al., Breast Cancer Res. Treat. 24: 85-95 (1992); Pietras et al., Oncogene 10: 2435-2446 (1995); and Donovan et al., J. Biol. Chem. 276: 40888-40895 (2001)). Since these RTKs activate Src and given observations that Src impairs the Cdk2 inhibitory action of p27, Src inhibition could restore antiestrogen responsiveness to resistant cells; AZD0530 is a potent, orally available dual inhibitor of AbI and Src family kinases (see Pie et al., J. Med. Chem. 47: 871-887 (2004)). It shows potent (<mM IC50) antiproliferative activity in 17 human cancer cell lines and inhibited growth of 6 of 13 xenograft tumor models tested. AZD0530 inhibits cell motility and invasion in vitro (see Hiscox et al., Breast Cancer Res. Treat. 97: 263-274 (2005)) and inhibits metastasis in animal models in vivo (see T. Green et al., 2005, American Association for Cancer Research, abstract). In three different Src-activated ER-positive breast cancer lines, AZD0530 or ER-saturating doses of tamoxifen each, when given alone, caused partial cell cycle inhibition. AZD0530 together with tamoxifen increased p27, inhibited cyclin E- Cdk2, and causedGl arrest. In contrast, in lines lacking Src activation, addition of AZD0530 did not enhance the antiproliferative effect of tamoxifen.

[0031] Analysis shows a statistical association between low nuclear p27 and Src activation in 482 primary human breast cancers (see Chu et al., Cell 128: 281-294 (2007)). Of 392 ER- positive cancers assayed, 127 of 339 (37%) showed Src activity scores of 2-3. This proportion is similar to the fraction of ER-positive breast cancers that manifest de novo antiestrogen resistance (see Chu, ibid.). These data raise the provocative possibility that Src inhibitors could restore response to tamoxifen in breast cancers with Src activation. The present data not only identify a novel mechanism whereby RTKs via Src promote p27 phosphorylation and proteolysis, they also provide a rationale for testing Src inhibitors together with antiestrogens in xenografts and to further explore their therapeutic potential in human breast cancer. Ubiquitin agents, such as the ubiquitin activating agents, ubiquitin conjugating agents, and ubiquitin ligating agents, are key determinants not only in ubiquitin-mediated proteolytic pathway that results in the degradation of targeted proteins, but also in regulation of cellular processes. Consequently, agents that modulate the activity of such ubiquitin agents may be used to up-regulate or down-regulate specific molecules involved in cellular signal transduction. Disease processes can be treated by such up or down regulation of signal transducers to enhance or dampen specific cellular responses. This principle has been used in the design of a number of therapeutics, including phosphodiesterase inhibitors for airway disease and vascular insufficiency, kinase inhibitors for malignant transformation and proteasome inhibitors for inflammatory conditions such as arthritis. [0032] Ubiquitin agents are involved in cancer, inflammation, adaptive immunity, innate immunity, bone metabolism, LPS-induced angiogenesis, osteoporosis, osteopinneal diseases, protozoan infection, viral infections, lymph node development, mammary gland development, skin development, and central nervous system development. Viral and protozoan infections involving ubiquitination include infections caused by variola viruses such as smallpox, HIV and related conditions, human papillomavirus, HSV, adenovirus, coxsackie virus, HCMV, KSHV, EBV, paramyxovirus, myxomavirus, ebola, retrovirus, rhabdovirus, and the malaria parasite. Other conditions in which ubiquitin is involved include aberrant cell growth, cancers, restenosis, psoriasis, and neoplastic cell proliferation. Ubiquitin is also involved in gene regulation, such as in bone metabolism, and in signal transduction pathways that involve IL-I, CD40, RANKL, LPS, IL-17, LMPl, NF-kB, AP-I and kinases, such as MAP kinases, JNK/SAPK, ERK, p38, IkB kinase and Src-family tyrosine kinases. Further, ubiquitin plays a critical role in the TNFR/IL-IR/TLR signal transduction pathways of inflammation, for example, in autoimmune diseases such as rheumatoid arthritis (RA), chronic obstructive pulmonary disease (COPD), inflammatory bowel disease (IBD) and graft rejection, bone-destructive diseases, such as osteoporosis and RA, allergies and infective disease such as bacterial sepsis and associated systemic inflammation. Finally, ubiquitin plays a role in diseases and conditions that involve non-degradative ubiquitination, for example, in diseases and conditions that involve activation of K-63 linked, non-degradative ubiquitination.

[0033] Ubiquitin has also been implicated as key components in other biochemical processes. Ubiquitination of the Gag structural protein of Rous Sarcoma virus has been linked to the targeting of Gag to the cell membrane of the host cell where it can assemble into spherical particles and bud from the cell surface. Production of HIV particles has also been associated with ubiquitination and may constitute an important cellular pathway for producing infectious particles. Thus, the ubiquitin pathway may be an important target for treatment of HIV positive patients.

[0034] Due to the importance of ubiquitin-mediated proteolysis and regulation of cellular processes, for example cell cycle regulation and developmental processes and, consequently, disease states, there is a need for compounds that inhibit ubiquitin ligases. Such inhibitors can be used to inhibit and treat such diseases, and as research tools to identify the physiological role of ubiquitin ligases in various regulatory pathways and disease states.

BRIEF SUMMARY OF THE INVENTION

[0035] The invention comprises compounds and compositions comprising the compounds for inhibiting ubiquitin agents. The compositions can further comprise a pharmaceutically acceptable carrier, diluent, and/or excipient and can be used in inhibiting and treating various conditions where ubiquitination is involved. They can also be used as research tools to study the role of ubiquitin in various natural and pathological processes.

[0036] In a first aspect, the invention comprises compounds that inhibit ubiquitination of target proteins.

[0037] In a second aspect, the invention comprises a pharmaceutical composition comprising an inhibitor of ubiquitination according to the invention and a pharmaceutically acceptable carrier, excipient, or diluent.

[0038] In a third aspect, the invention comprises methods of inhibiting ubiquitination in a cell, comprising contacting a cell in which inhibition of ubiquitination is desired with a pharmaceutical composition comprising a ubiquitin agent inhibitor according to the invention.

[0039] In a fourth aspect, the invention provides methods for treating cell proliferative diseases or conditions, comprising administering to a patient in need thereof a pharmaceutical composition comprising an effective amount of a ubiquitin agent inhibitor according to the invention.

[0040] In a fifth aspect, the invention provides methods for inhibiting TRAF6 activity in a cell, comprising administering to the cell a compound of the invention or a pharmaceutical composition comprising an effective amount of a compound according to the invention.

[0041] The foregoing only summarizes certain aspects of the invention and is not intended to be limiting in nature. These aspects and other aspects and embodiments are described more fully below. All patent applications and publications of any sort referred to in this specification are hereby incorporated by reference in their entirety. In the event of a discrepancy between the express disclosure of this specification and a patent application or publication incorporated by reference, the express disclosure of this specification shall control.

BRIEF DESCRIPTION OF THE DRAWINGS

[0042] Figure 1 describes the Protein Ubiquitination System

[0043] Figure 2 describes Phosphorylation- and ubiquitin-dependent degradation of p27

[0044] Figure 3 describes Ubiquitination of p27Kipl by SCF-Skp2

[0045] Figure 4 describes a Plate -based p27-SCF ubiquitination assay

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0046] The invention provides compounds, compositions and methods for inhibiting ubiquitin ligase activity. In particular, the invention provides compounds, compositions and methods for inhibiting TRAF6, APC as well as other enzymes that exhibit E3-like activity. The invention also provides methods and compositions for treating cell proliferative diseases and conditions in which TRAF6 is involved.

[0047] The compounds of the invention inhibit the ubiquitination of p27, a tumor suppressor, and biochemical and cell based assays. The compounds are expected to show selectivity over ligase counter assays such as p53-MDM2, APC, TRAF 6 and other biological targets and biochemical and cell based assays. The compounds of the invention are expected to have good oral bioavailability, good ADME properties and are readily amenable to oral or IV formulation. The compounds of the invention are expected to show efficacy in vivo graft animal models in diseases mediated via pathways involving p27/SCF. [0048] The invention provides compounds according Formula I:

and pharmaceutically acceptable salts, hydrates, solvates, polymorphs, atrophisomers, N-oxides, and prodrugs thereof, wherein

L' is a covalent bond or -SO₂-;

X is H or -OH

Ri and R₂ are independently -H, -NO₂, -OH, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, mono- to per-halogenated Ci-C₆ alkyl, heterocyclyl, heterocyclyl-O-aryl, -NH₂, mono- or (Ii-(C₁-C₆ alkyl) amino, -C(O)-OR₉, -N(R₈)-Z, -S-Z, -SO₂-Z, -N(R₈)-Z, aryl, or heteroaryl, wherein each of the aryl and heteroaryl is optionally substituted with 1 to 3 groups selected from -H, Ci-C₆ alkyl, Ci-C₆ alkoxy, mono- to per-halogenated Ci-C₆ alkyl, -NH₂, mono- or di-(Ci-C₆ alkyl) amino, -NO₂, halo, or -CN; or

Ri and R₂ together with the carbon atoms to which they are attached form an aryl group optionally substituted with 1 to 3 groups selected from -OH, halo, Ci-C₆-alkyl, Ci-C₆-alkoxy, -NH₂. and -NO₂;

Y is -H, -NO₂, -OH, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, -NH₂, mono- or (Ii-(C₁-C₆ alkyl) amino, -C(O)-OR₉, -C(O)R₉, -N(R₈)-Z, Ci-C₆ alkyl-aryl, Ci-C₆ alkyl-heterocyclyl, Ci-C₆ alkyl-heteroaryl, C₀-C₆ alkyl-C(O)-aryl, C₀-C₆ alkyl-C(O)-heterocyclyl, C₀-C₆ alkyl-C(O)-heteroaryl, -O-aryl, aryl, heterocyclyl, heteroaryl, or -Zi-S(O)₂-Z₂ where Zi and Z₂ are independently aryl or heteroaryl, wherein each of the aryl, heterocyclyl and heteroaryl is optionally substituted with 1 to 3 groups selected from -H, Ci-C₆ alkyl, Ci-C₆ alkoxy, mono- to per-halogenated Ci-C₆ alkyl, -C(O)-OR₉, -N(R₉)-C(O)R₉, -O-(halo C₁-C₆ alkyl), -NH₂, mono- or di-(Ci-C₆ alkyl) amino, -NO₂, halo, or -CN;

R₈ is -H, Ci-C₆ alkyl, or Ci-C₆ alkoxy;

R₉ is -H, Ci-C₆ alkyl or C₃-C₆ cycloalkyl; and

Z is -Ci-C₆ alkyl-O-C(O)-Ci-C₆ alkyl, aryl or heteroaryl wherein each of the aryl and heteroaryl is optionally substituted with -H, -NO₂, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, -NH₂, mono- or di-(Ci-C₆ alkyl) amino, or -C(O)-OR₉. [0049] In a particular embodiment, the compounds of formula I are compounds of formula Ia:

[0050] In a particular embodiment, the compounds of formula II are compounds of formulae III(i)-III(vi)

IΙI(i) X is H;

IΙI(ii) X is -OH;

IΙI(iii) X is H and Y is optionally substituted aryl;

IΙI(iv) X is OH and Y is optionally substituted aryl;

III(v) X is H and Y is aryl substituted with halo (preferably -Cl); or

IΙI(vi) X is OH and Y is aryl substituted with halo (preferably -Cl). [0051] In a particular embodiment, the compounds according to formula I, II, and III(i)-III(vi) are compounds of formulae IV(i)-IV(iii):

IV(i) Ri is -S-Z;

IV(ii) Ri is -SO₂-Z; or

IV(iii) Ri is -N(R₈)-Z.

[0052] In a particular embodiment, the compounds according to formula I, II, III(i)-III(vi), and IV(i)-IV(iii) are compounds of formula V, which are compounds in which Z is aryl or heteroaryl wherein each of the aryl and heteroaryl is optionally substituted with -H, -NO₂, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, -NH₂, mono- or di-(Ci-C₆ alkyl) amino, or -C(O)-ORg. In preferred embodiments of compounds of formula V, Z is phenyl, pyrimidinyl, quinolinyl, or piperidinyl each of which is unsubstituted or substituted as described in the definition of Z in the description of compounds of formula I. Of the compounds in which Z is pyrimidinyl, preferred are those compounds in which Z is pyrimidin-4-yl. Of the compounds in which Z is quinolinyl, preferred are those compounds in which Z is quinolinyl-2-yl or quinolinyl-8-yl. [0053] In a particular embodiment, the compounds according to formula I, II, III(i)-III(vi), IV(i)-IV(iii), and V are compounds of formula VI, which are compounds in which R₂ is H. [0054] Preferred compounds according to the invention for those of Table 1 :

[0055] Specifically excluded from the scope of the invention are compounds of Tables 2 and

2A:

Table 2A

Table 2A

Table 2A

Table 2A

Table 2A

Table 2A

Table 2A

Table 2A

Table 2A

Table 2A

Table 2A

Table 2A

[0056] For the purposes of the United States, the invention also comprises compounds of Table 1, but the compounds of Table 1 are specifically excluded from the scope of the compounds of formulae I-V.

[0057] The compounds according to the first aspect of the invention are also useful as general ubiquitin ligase inhibitors. For example, the compounds of the invention can be used as inhibitors of enzymes that exhibit ligase activity, including but not limited to TRAF6, APC and E3 enzymes. The compounds of the invention are also useful for regulating or inhibiting pathways in diseases and conditions that involve ubiquitin conjugation and deconjugation such as cancers, inflammation, metabolism, viral diseases and central nervous system disorders. For example, the compounds of the invention can be used to regulate or inhibit the products of genes that encode ubiquitin or ubiquitin- like enzymes described in Wong et al. (DDT 8 (16), 746-754 (2003)), which is incorporated by reference in its entirety.

[0058] In the second aspect, the invention provides for pharmaceutical compositions comprising, together with a pharmaceutically acceptable carrier, diluent, or excipient, a compound of according to the first aspect of the invention described above. [0059] In the third aspect, the invention provides methods of inhibiting ubiquitination in a cell comprising contacting the cell in which inhibition of ubiquitination is desired with a compound according to the first aspect of the invention or a pharmaceutical composition according to the second aspect of the invention. The compounds and formulations of the invention can inhibit ubiquitination in cells derived from animals, particularly, mammalian cells. The compounds and formulations of the invention can also be used to inhibit the ubiquitin ligase activity of TRAF6.

[0060] In the fourth aspect, the invention provides for methods of treating cell proliferative diseases or conditions comprising administering to a patient an effective amount of a compound of the first aspect of the invention or a pharmaceutical composition according to the second aspect of the invention. Cell proliferative diseases or conditions include, but are not limited to, psoriasis, keloid scarring, and cancers, such as cancers of the breast, immune system, bone, nervous system, brain, blood, lymphatic system, and skin. Particularly, the compounds and pharmaceutical compositions of the invention are useful for treating cell proliferative diseases or conditions that involve TRAF6.

[0061] In the fifth aspect, the invention provides for methods of inhibiting TRAF6 comprising administering to a patient an effective amount of a compound according to the first aspect of the invention or pharmaceutical composition according to the second aspect of the invention. Particularly, the compounds and pharmaceutical compositions are useful for treating conditions or diseases that involve TRAF6 such as those related to cancer, inflammation, adaptive immunity, innate immunity, bone metabolism, LPS-induced angiogenesis, osteoporosis, osteopinneal diseases, lymph node development, mammary gland development, skin development, and central nervous system development.

[0062] The compounds for use in the method according to the fifth aspect of the invention are also useful as general ubiquitin ligase inhibitors. For example, the compounds of the invention can be used as inhibitors of E3 enzymes that contain HECT and RING finger domains, Mdm2 with RING fingers and variants, and U-box-containing proteins. Accordingly, the compounds of the invention are useful as protein modulators, immunologic agents anti-inflammatory agents, anti-osteoporosis agents, anti-viral agents, for example, inhibitors of variola viruses such as smallpox, HIV and related conditions, human papillomavirus, HSV, adenovirus, coxsackie virus, HCMV, KSHV, EBV, paramyxovirus, myxomavirus, ebola, retrovirus, and rhabdovirus, anti-protozoan agents, for example, inhibitors of the malaria parasite. The compounds of the invention are also useful as oncologic and anti-pro liferative agents that inhibit aberrant cell growth, cancers, restenosis, psoriasis, and neoplastic cell proliferation.

[0063] Inhibition of TRAF 6 activity by the compounds and pharmaceutical compositions of the invention provides ways to regulate the expression of genes involved many biological processes. Such processes include but are not limited to bone metabolism and signal transduction pathways that involve IL-I, CD40, RANKL, LPS, IL-17, and LMPl. For example, the compounds and pharmaceutical compositions of the invention can be used to regulate the activities of transcription factors that activate the expression of genes, such as NF -kB and AP-I. The compounds and pharmaceutical compositions can also be used to regulate the activities of kinases, such as MAP kinases, JNK/SAPK, ERK, p38, IkB kinase and Src-family tyrosine kinases.

[0064] Inhibition of TRAF6 serves as a therapeutic target for inflammatory and autoimmune diseases. For example, TRAF6 plays a critical regulator role of the TNFR/IL-1R/TLR signal transduction pathways and can serve as a broad anti-inflammation target for inflammatory diseases such as RA, COPD, IBD. Further TRAF6 can also be a useful therapeutic target for treating autoimmune diseases, such as graft rejection because of its regulator role in the CD40 signaling cascade. TRAF6 is also a target for treating bone-destructive diseases, such as osteoporosis and rheumatoid arthritis because TRAF6 plays a critical regulator role of the RANK signal transduction that mediate osteoclast activation and function. Similarly, TRAF6 may also serve as a novel allergic and infective disease target for treating bacterial sepsis and associated systemic inflammation because TRAF6 plays critical mediator roles in the TLR signal transduction which is involved in the interaction between dentritic cells, T lymphocytes and mast cells.

[0065] Inhibition of TRAF 6 may also serve as a therapeutic target in diseases and conditions that involve non-degradative ubiquitination. For example, TRAF6 acts as an E3 ubiquitin ligase that mediates kinase activation by K-63 linked, non-degradative ubiquitination. Inhibiting TRAFβ's E3 ligase activity may provide novel anti-inflammation therapeutics. [0066] In a sixth aspect, the invention provides compounds according Formula X:

(X) and pharmaceutically acceptable salts, hydrates, solvates, polymorphs, atrophisomers,

N-oxides, and prodrugs thereof, wherein n is 0, 1, 2, 3, or 4; L' is a covalent bond or -SO₂-; A is -C(X)(Y)-, -N(Y)-, or -O-, wherein X is H, -C(O)R₈, or -OH; and

Y is -H, -NO₂, -OH, -CN, halo, -Z, Ci-C₆ alkyl, Ci-C₆ alkoxy, -NH₂, mono- or (Ii-(C₁-C₆ alkyl) amino, -C(O)-OR₉, -C(O)R₉, -N(R₈)-Z, Ci-C₆ alkyl-aryl, Ci-C₆ alkyl-heterocyclyl, C₁-C₆ alkyl-heteroaryl, C₀-C₆ alkyl-C(O)-aryl, C₀-C₆ alkyl-C(O)-heterocyclyl, C₀-C₆ alkyl-C(O)-heteroaryl, -O-aryl, C₃-C₈-cycloalkyl, heterocyclyl, , or -Zi-S(O)₂-Z₂ where

Zi and Z₂ are independently aryl or heteroaryl, wherein each of the aryl, heterocyclyl and heteroaryl is optionally substituted with 1 to 3 groups selected from -H, Ci-C₆ alkyl, Ci-C₆ alkoxy, mono- to per-halogenated Ci-C₆ alkyl, -C(O)-OR₉, -N(R₉)-C(O)R₉, -O-(halo Ci-C₆ alkyl), -NH₂, mono- or di-(Ci-C₆ alkyl) amino, -NO₂, halo, or -CN; or X and Y together with the atom to which they are attached, are oxo, a C₃-Cs cycloalkyl or heterocyclyl wherein the cycloalkyl or heterocyclyl is optionally substituted by one to four R⁴ groups;

Ri and R₂ are independently -H, -NO₂, -OH, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, mono- to per-halogenated Ci-C₆ alkyl, heterocyclyl, heterocyclyl-O-aryl, -NH₂, mono- or (Ii-(C₁-C₆ alkyl) amino, -C(O)-OR₉, -N(R₈)-Z, -0-Z, -S-Z, -SO₂-Z, -SO₂-NH-, -N(Rg)-Z, aryl, or heteroaryl, wherein each of the aryl and heteroaryl is optionally substituted with 1 to 3 groups selected from -H, Ci-C₆ alkyl, Ci-C₆ alkoxy, mono- to per-halogenated Ci-C₆ alkyl, -NH₂, mono- or di-(Ci-C₆ alkyl) amino, -NO₂, halo, or -CN; or Ri and R₂ together with the carbon atoms to which they are attached form an aryl group optionally substituted with 1 to 3 groups selected from -OH, halo, Ci-C₆-alkyl, Ci-C₆-alkoxy, -NH₂. and -NO₂; each R₃ is independently halo or Ci-C₆ alkyl, or two R₃ attached to adjacent carbon atoms, together with the atoms to which they are attached, form a fused aryl or heteroaryl wherein the aryl or heteroaryl is optionally substituted by one to four R⁴ groups; or Y and R₃ attached to adjacent carbon atoms, together with the atoms to which they are attached, form a fused aryl or heteroaryl and X is absent, wherein the aryl or heteroaryl is optionally substituted by one to four R⁴ groups; each R₄ is independently halo, Ci-C₆ alkyl, oxo, or Ci-C₆ alkoxy; Rs is -H, Ci-C₆ alkyl, or Ci-C₆ alkoxy; R₉ is -H, Ci-C₆ alkyl or C₃-C₆ cycloalkyl; and each Z is independently -Ci-C₆ alkyl-O-C(O)-Ci-C₆ alkyl, aryl, or heteroaryl wherein each of the aryl and heteroaryl is optionally substituted with one to four R₅ groups, wherein each R₅ is independently -NO₂, -CN, halo, C₁-C₆ alkyl, C₁-C₆ alkoxy, -NH₂, mono- or di-(Ci-C₆ alkyl) amino, oxo, or -C(O)ORg, provided the compound is not a compound listed in Tables 2 and 2A. [0067] In preferred embodiments of the compounds of formula X, L' is SO₂. [0068] In other preferred embodiments of the compounds of formula X, n is 0, 1, or 2. More preferably, n is 0. In other more preferred embodiments, n is 2.

[0069] In another preferred embodiment of compounds of formula X, A is -C(X)(Y)- wherein X is H or -OH; and Y is aryl, or heteroaryl wherein the aryl and heteroaryl is optionally substituted with one to four R5 groups, wherein each R5 is independently -NO₂, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, -NH₂, mono- or di-(Ci-C₆ alkyl) amino, or -C(O)ORg.

[0070] More preferred are compounds of formula X, wherein A is -CH(Y)- wherein

Y is aryl, or heteroaryl wherein the aryl and heteroaryl is optionally substituted with one to four R5 groups, wherein each R₅ is independently -NO₂, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, -NH₂, mono- or di-(Ci-C₆ alkyl) amino, or -C(O)OR₉.

[0071] Even more preferred are compounds of formula X, wherein A is -CH(Y)- wherein Y is aryl, optionally substituted with one or two R₅ groups, wherein each R₅ is independently -NO₂, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, -NH₂, mono- or di-(Ci-C₆ alkyl) amino, or -C(O)OR₉. [0072] In another preferred embodiment of compounds of formula X, A is -N(Y)-, wherein

[0073] More preferred are compounds of formula X, wherein A is -N(Y)- wherein

Y is aryl, optionally substituted with one or two R₅ groups, wherein each R₅ is independently -NO₂, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, -NH₂, mono- or di-(Ci-C₆ alkyl) amino, or -C(O)OR₉.

[0074] Other preferred compounds of formula X are those wherein A is -N(Y)- wherein Y is heteroaryl, optionally substituted with one or two R₅ groups, wherein each R₅ is independently

-NO₂, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, -NH₂, mono- or di-(Ci-C₆ alkyl) amino, or

-C(O)OR₉.

[0075] Even more preferred are compounds of formula X, wherein A is -N(Y)- wherein Y is phenyl, optionally substituted with one or two R₅ groups, wherein each R₅ is independently

-C(O)OR₉.

[0076] Other even more preferred are compounds of formula X, wherein A is -N(Y)- wherein Y is pyridyl, optionally substituted with one or two R₅ groups, wherein each R₅ is independently -NO₂, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, -NH₂, mono- or di-(Ci-C₆ alkyl) amino, or -C(O)OR₉.

[0077] In another preferred embodiment of compounds of formula X, Ri is -H, halo, -O-Z, -S-Z, -SO₂-Z, -N(Rg)-Z, aryl, or heteroaryl, wherein each of the aryl and heteroaryl is optionally substituted with 1 to 3 groups selected from -H, Ci-C₆ alkyl, Ci-C₆ alkoxy, mono- to per-halogenated Ci-C₆ alkyl, -NH₂, mono- or di-(Ci-C₆ alkyl) amino, -NO₂, halo, or -CN. [0078] More preferred are compounds of formula X, wherein Ri is -H, halo, -O-Z, -S-Z, or N(Rg)-Z. Even more preferred are compounds of formula X, wherein Ri is -O-Z, -S-Z, or N(Rg)-Z. Other more preferred are compounds of formula X, wherein Ri is -H, or halo. Other more preferred are compounds of formula X, wherein Ri is chloro.

[0079] In another preferred embodiment of compounds of formula X, Ri is -S-Z, wherein Z is aryl or heteroaryl (preferably, quinolinyl) wherein the aryl and heteroaryl is optionally substituted with one to four R5 groups, wherein each R5 is independently -NO₂, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, -NH₂, mono- or di-(Ci-C₆ alkyl) amino, or -C(O)OR₉. More preferred are compounds of formula X, wherein Ri is -S-Z, wherein Z is aryl optionally substituted with one or two R5 groups, wherein each R₅ is independently -NO₂, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, -NH₂, mono- or di-(Ci-C₆ alkyl) amino, or -C(O)OR₉. Even more preferred are compounds of formula X, wherein Ri is -S-Z, wherein Z is phenyl optionally substituted with one or two R₅ groups, wherein each R₅ is independently -NO₂, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, -NH₂, mono- or di-(Ci-C₆ alkyl) amino, or -C(O)OR₉.

[0080] In another preferred embodiment of compounds of formula X, Ri is -S-Z, wherein Z is heteroaryl (preferably, quinolinyl) optionally substituted with one or two R₅ groups, wherein each R₅ is independently -NO₂, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, -NH₂, mono- or di-(Ci-C₆ alkyl) amino, or -C(O)OR₉. Even more preferred are compounds of formula X, wherein Ri is -S-Z, wherein Z is pyridyl optionally substituted with one or two R₅ groups, wherein each R₅ is independently -NO₂, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, -NH₂, mono- or di-(Ci-C₆ alkyl) amino, or -C(O)OR₉. Even more preferred are compounds of formula X, wherein Ri is -S-Z, wherein Z is quinolinyl, optionally substituted with one or two R₅ groups, wherein each R₅ is independently -NO₂, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, -NH₂, mono- or CIi-(C₁-C₆ alkyl) amino, or -C(O)OR₉. [0081] In another preferred embodiment of compounds of formula X, R₂ is -H, -NO₂, -OH,

-CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, mono- to per-halogenated Ci-C₆ alkyl, -NH₂, mono- or

(Ii-(C₁-C₆ alkyl) amino, -C(O)-OR₉, -N(R₈)-Z, -O-Z, -S-Z, -SO₂-Z, or -N(R₈)-Z. More preferred are compounds of formula X, wherein R₂ is -H, -NO₂, -OH, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, mono- to per-halogenated Ci-C₆ alkyl, -NH₂, mono- or di-(Ci-C₆ alkyl) amino,

-C(O)-OR₉. Even more preferred are compounds of formula X, wherein R₂ is -H.

[0082] In another preferred embodiment of compounds of formula X, R₃ is methyl. More preferred compounds of formula X are those where R3 is methyl and n is O, 1, or 2.

[0083] In a particular embodiment, the compounds of formula X are compounds of formula

XI:

XL

[0084] In preferred embodiments of the compounds of formula XI, L' is SO₂. [0085] In other preferred embodiments of the compounds of formula XI, n is 0, 1, or 2. More preferably, n is 0. In other more preferred embodiments, n is 2. [0086] In another preferred embodiment of compounds of formula XI, A is -C(X)(Y)- wherein X is H or -OH; and Y is aryl, or heteroaryl wherein the aryl and heteroaryl is optionally substituted with one to four R₅ groups, wherein each R₅ is independently -NO₂, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, -NH₂, mono- or (Ii-(C₁-C₆ alkyl) amino, or -C(O)OR₉. [0087] More preferred are compounds of formula XI, wherein A is -CH(Y)- wherein

Y is aryl, or heteroaryl wherein the aryl and heteroaryl is optionally substituted with one to four R₅ groups, wherein each R₅ is independently -NO₂, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, -NH₂, mono- or di-(Ci-C₆ alkyl) amino, or -C(O)OR₉.

[0088] Even more preferred are compounds of formula XI, wherein A is -CH(Y)- wherein

[0089] In another preferred embodiment of compounds of formula XI, A is -N(Y)-, wherein Y is aryl, or heteroaryl wherein the aryl and heteroaryl is optionally substituted with one to four R₅ groups, wherein each R₅ is independently -NO₂, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, -NH₂, mono- or di-(Ci-C₆ alkyl) amino, or -C(O)ORg.

[0090] More preferred are compounds of formula XI, wherein A is -N(Y)- wherein

[0091] Other preferred compounds of formula XI are those wherein A is -N(Y)- wherein

Y is heteroaryl, optionally substituted with one or two R₅ groups, wherein each R₅ is independently -NO₂, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, -NH₂, mono- or di-(Ci-C₆ alkyl) amino, or -C(O)OR₉.

[0092] Even more preferred are compounds of formula XI, wherein A is -N(Y)- wherein

Y is phenyl, optionally substituted with one or two R₅ groups, wherein each R₅ is independently -NO₂, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, -NH₂, mono- or di-(Ci-C₆ alkyl) amino, or -C(O)OR₉.

[0093] Other even more preferred are compounds of formula XI, wherein A is -N(Y)- wherein Y is pyridyl, optionally substituted with one or two R₅ groups, wherein each R₅ is independently -NO₂, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, -NH₂, mono- or di-(Ci-C₆ alkyl) amino, or -C(O)OR₉.

[0094] In another preferred embodiment of compounds of formula XI, Ri is -H, halo, -O-Z, -S-Z, -SO₂-Z, -N(Rg)-Z, aryl, or heteroaryl, wherein each of the aryl and heteroaryl is optionally substituted with 1 to 3 groups selected from -H, Ci-C₆ alkyl, Ci-C₆ alkoxy, mono- to per-halogenated Ci-C₆ alkyl, -NH₂, mono- or di-(Ci-C₆ alkyl) amino, -NO₂, halo, or -CN. [0095] More preferred are compounds of formula XI, wherein Ri is -H, halo, -O-Z, -S-Z, or N(Rg)-Z. Even more preferred are compounds of formula XI, wherein Ri is -O-Z, -S-Z, or N(Rg)-Z. Other more preferred are compounds of formula XI, wherein Ri is -H, or halo. Other more preferred are compounds of formula XI, wherein Ri is chloro.

[0096] In another preferred embodiment of compounds of formula XI, Ri is -S-Z, wherein Z is aryl or heteroaryl (preferably, quinolinyl) wherein the aryl and heteroaryl is optionally substituted with one to four R₅ groups, wherein each R₅ is independently -NO₂, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, -NH₂, mono- or di-(Ci-C₆ alkyl) amino, or -C(O)OR₉. More preferred are compounds of formula XI, wherein Ri is -S-Z, wherein Z is aryl optionally substituted with one or two R₅ groups, wherein each R₅ is independently -NO₂, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, -NH₂, mono- or di-(Ci-C₆ alkyl) amino, or -C(O)ORg. Even more preferred are compounds of formula XI, wherein Ri is -S-Z, wherein Z is phenyl optionally substituted with one or two R₅ groups, wherein each R₅ is independently -NO₂, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, -NH₂, mono- or di-(Ci-C₆ alkyl) amino, or -C(O)ORg.

[0097] In another preferred embodiment of compounds of formula XI, Ri is -S-Z, wherein Z is heteroaryl (preferably, quinolinyl) optionally substituted with one or two R₅ groups, wherein each R₅ is independently -NO₂, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, -NH₂, mono- or di-(Ci-C₆ alkyl) amino, or -C(O)ORg. Even more preferred are compounds of formula XI, wherein Ri is -S-Z, wherein Z is pyridyl optionally substituted with one or two R₅ groups, wherein each R₅ is independently -NO₂, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, -NH₂, mono- or di-(Ci-C₆ alkyl) amino, or -C(O)ORg. Even more preferred are compounds of formula XI, wherein Ri is -S-Z, wherein Z is quinolinyl, optionally substituted with one or two R₅ groups, wherein each R₅ is independently -NO₂, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, -NH₂, mono- or di-(Ci-C₆ alkyl) amino, or -C(O)ORg.

[0098] In another preferred embodiment of compounds of formula XI, R₂ is -H, -NO₂, -OH, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, mono- to per-halogenated Ci-C₆ alkyl, -NH₂, mono- or di-(Ci-C₆ alkyl) amino, -C(O)-OR₉, -N(Rg)-Z, -0-Z, -S-Z, -SO₂-Z, or -N(Rg)-Z. More preferred are compounds of formula XI, wherein R₂ is -H, -NO₂, -OH, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, mono- to per-halogenated Ci-C₆ alkyl, -NH₂, mono- or di-(Ci-C₆ alkyl) amino, -C(O)-ORg. Even more preferred are compounds of formula XI, wherein R₂ is -H. [0099] In another preferred embodiment of compounds of formula XI, R₃ is methyl. More preferred compounds of formula XI are those where R₃ is methyl and n is O, 1 , or 2. [0100] In a particular embodiment, the compounds of formula XI are compounds of formula XII:

XII. [0101] In other preferred embodiments of the compounds of formula XII, n is 0, 1, or 2.

More preferably, n is 0. In other more preferred embodiments, n is 2.

[0102] In another preferred embodiment of compounds of formula XII, A is -C(X)(Y)- wherein X is H or -OH; and Y is aryl, or heteroaryl wherein the aryl and heteroaryl is optionally substituted with one to four R5 groups, wherein each R5 is independently -NO₂, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, -NH₂, mono- or di-(Ci-C₆ alkyl) amino, or -C(O)ORg.

[0103] More preferred are compounds of formula XII, wherein A is -CH(Y)- wherein Y is aryl, or heteroaryl wherein the aryl and heteroaryl is optionally substituted with one to four R₅ groups, wherein each R₅ is independently -NO₂, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, -NH₂, mono- or di-(Ci-C₆ alkyl) amino, or -C(O)OR₉.

[0104] Even more preferred are compounds of formula XII, wherein A is -CH(Y)- wherein Y is aryl, optionally substituted with one or two R₅ groups, wherein each R₅ is independently -NO₂,

-CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, -NH₂, mono- or di-(Ci-C₆ alkyl) amino, or -C(O)OR₉.

[0105] In another preferred embodiment of compounds of formula XII, A is -N(Y)-, wherein

Y is aryl, or heteroaryl wherein the aryl and heteroaryl is optionally substituted with one to four

R5 groups, wherein each R₅ is independently -NO₂, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, -NH₂, mono- or di-(Ci-C₆ alkyl) amino, or -C(O)OR₉.

[0106] More preferred are compounds of formula XII, wherein A is -N(Y)- wherein Y is aryl, optionally substituted with one or two R₅ groups, wherein each R₅ is independently -NO₂,

[0107] Other preferred compounds of formula XII are those wherein A is -N(Y)- wherein Y is heteroaryl, optionally substituted with one or two R₅ groups, wherein each R₅ is independently

-C(O)OR₉.

[0108] Even more preferred are compounds of formula XII, wherein A is -N(Y)- wherein Y is phenyl, optionally substituted with one or two R₅ groups, wherein each R₅ is independently

-C(O)OR₉.

[0109] Other even more preferred are compounds of formula XII, wherein A is -N(Y)- wherein Y is pyridyl, optionally substituted with one or two R₅ groups, wherein each R₅ is independently -NO₂, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, -NH₂, mono- or di-(Ci-C₆ alkyl) amino, or -C(O)OR₉.

[0110] In another preferred embodiment of compounds of formula XII, Ri is -H, halo, -O-Z, -S-Z, -SO₂-Z, -N(Rg)-Z, aryl, or heteroaryl, wherein each of the aryl and heteroaryl is optionally substituted with 1 to 3 groups selected from -H, Ci-C₆ alkyl, Ci-C₆ alkoxy, mono- to per-halogenated Ci-C₆ alkyl, -NH₂, mono- or di-(Ci-C₆ alkyl) amino, -NO₂, halo, or -CN. [0111] More preferred are compounds of formula XII, wherein Ri is -H, halo, -O-Z, -S-Z, or N(Rg)-Z. Even more preferred are compounds of formula XII, wherein Ri is -O-Z, -S-Z, or N(Rg)-Z. Other more preferred are compounds of formula XII, wherein Ri is -H, or halo. Other more preferred are compounds of formula XII, wherein Ri is chloro.

[0112] In another preferred embodiment of compounds of formula XII, Ri is -S-Z, wherein Z is aryl or heteroaryl (preferably, quinolinyl) wherein the aryl and heteroaryl is optionally substituted with one to four R5 groups, wherein each R5 is independently -NO₂, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, -NH₂, mono- or di-(Ci-C₆ alkyl) amino, or -C(O)OR₉. More preferred are compounds of formula XII, wherein Ri is -S-Z, wherein Z is aryl optionally substituted with one or two R5 groups, wherein each R₅ is independently -NO₂, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, -NH₂, mono- or di-(Ci-C₆ alkyl) amino, or -C(O)OR₉. Even more preferred are compounds of formula XII, wherein Ri is -S-Z, wherein Z is phenyl optionally substituted with one or two R₅ groups, wherein each R₅ is independently -NO₂, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, -NH₂, mono- or di-(Ci-C₆ alkyl) amino, or -C(O)OR₉.

[0113] In another preferred embodiment of compounds of formula XII, Ri is -S-Z, wherein Z is heteroaryl (preferably, quinolinyl) optionally substituted with one or two R₅ groups, wherein each R₅ is independently -NO₂, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, -NH₂, mono- or di-(Ci-C₆ alkyl) amino, or -C(O)OR₉. Even more preferred are compounds of formula XII, wherein Ri is -S-Z, wherein Z is pyridyl optionally substituted with one or two R₅ groups, wherein each R₅ is independently -NO₂, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, -NH₂, mono- or di-(Ci-C₆ alkyl) amino, or -C(O)OR₉. Even more preferred are compounds of formula XII, wherein Ri is -S-Z, wherein Z is quinolinyl optionally substituted with one or two R₅ groups, wherein each R₅ is independently -NO₂, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, -NH₂, mono- or di-(Ci-C₆ alkyl) amino, or -C(O)OR₉. [0114] In another preferred embodiment of compounds of formula XII, R₂ is -H, -NO₂, -OH,

(Ii-(C₁-C₆ alkyl) amino, -C(O)-OR₉, -N(R₈)-Z, -0-Z, -S-Z, -SO₂-Z, or -N(R₈)-Z. More preferred are compounds of formula XII, wherein R₂ is -H, -NO₂, -OH, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, mono- to per-halogenated Ci-C₆ alkyl, -NH₂, mono- or di-(Ci-C₆ alkyl) amino,

-C(O)-OR₉. Even more preferred are compounds of formula XII, wherein R₂ is -H.

[0115] In another preferred embodiment of compounds of formula XII, R₃ is methyl. More preferred compounds of formula XII are those where R3 is methyl and n is O, 1 , or 2.

[0116] In a particular embodiment, the compounds of formula XII are compounds of formula

XIII:

XIII.

[0117] In other preferred embodiments of the compounds of formula XIII, n is 0, 1, or 2. More preferably, n is 0. In other more preferred embodiments, n is 2. [0118] In another preferred embodiment of compounds of formula XIII, A is -C(X)(Y)- wherein X is H or -OH; and Y is aryl, or heteroaryl wherein the aryl and heteroaryl is optionally substituted with one to four R₅ groups, wherein each R₅ is independently -NO₂, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, -NH₂, mono- or di-(Ci-C₆ alkyl) amino, or -C(O)OR₉. [0119] More preferred are compounds of formula XIII, wherein A is -CH(Y)- wherein [0120] Y is aryl, or heteroaryl wherein the aryl and heteroaryl is optionally substituted with one to four R₅ groups, wherein each R₅ is independently -NO₂, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, -NH₂, mono- or di-(Ci-C₆ alkyl) amino, or -C(O)OR₉.

[0121] Even more preferred are compounds of formula XIII, wherein A is -CH(Y)- wherein Y is aryl, optionally substituted with one or two R₅ groups, wherein each R₅ is independently -NO₂, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, -NH₂, mono- or di-(Ci-C₆ alkyl) amino, or -C(O)OR₉.

[0122] In another preferred embodiment of compounds of formula XIII, A is -N(Y)-, wherein Y is aryl, or heteroaryl wherein the aryl and heteroaryl is optionally substituted with one to four R₅ groups, wherein each R₅ is independently -NO₂, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, -NH₂, mono- or di-(Ci-C₆ alkyl) amino, or -C(O)OR₉.

[0123] More preferred are compounds of formula XIII, wherein A is -N(Y)- wherein Y is aryl, optionally substituted with one or two R₅ groups, wherein each R₅ is independently -NO₂, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, -NH₂, mono- or (Ii-(C₁-C₆ alkyl) amino, or -C(O)OR₉. [0124] Other preferred compounds of formula XIII are those wherein A is -N(Y)- wherein Y is heteroaryl, optionally substituted with one or two R₅ groups, wherein each R₅ is independently -NO₂, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, -NH₂, mono- or di-(Ci-C₆ alkyl) amino, or -C(O)OR₉.

[0125] Even more preferred are compounds of formula XIII, wherein A is -N(Y)- wherein Y is phenyl, optionally substituted with one or two R₅ groups, wherein each R₅ is independently -NO₂, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, -NH₂, mono- or di-(Ci-C₆ alkyl) amino, or -C(O)OR₉.

[0126] Other even more preferred are compounds of formula XIII, wherein A is -N(Y)- wherein Y is pyridyl, optionally substituted with one or two R₅ groups, wherein each R₅ is independently -NO₂, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, -NH₂, mono- or di-(Ci-C₆ alkyl) amino, or -C(O)OR₉.

[0127] In another preferred embodiment of compounds of formula XIII, wherein Z is aryl or heteroaryl (preferably, quinolinyl) wherein the aryl and heteroaryl is optionally substituted with one to four R₅ groups, wherein each R₅ is independently -NO₂, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, -NH₂, mono- or di-(Ci-C₆ alkyl) amino, or -C(O)OR₉. More preferred are compounds of formula XIII, wherein Z is aryl optionally substituted with one or two R₅ groups, wherein each R₅ is independently -NO₂, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, -NH₂, mono- or di-(Ci-C₆ alkyl) amino, or -C(O)OR₉. Even more preferred are compounds of formula XIII, wherein Z is phenyl optionally substituted with one or two R₅ groups, wherein each R₅ is independently -NO₂, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, -NH₂, mono- or di-(Ci-C₆ alkyl) amino, or -C(O)OR₉. [0128] In another preferred embodiment of compounds of formula XIII, Z is heteroaryl (preferably, quinolinyl) optionally substituted with one or two R₅ groups, wherein each R₅ is independently -NO₂, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, -NH₂, mono- or di-(Ci-C₆ alkyl) amino, or -C(O)OR₉. Even more preferred are compounds of formula XIII, Z is pyridyl optionally substituted with one or two R₅ groups, wherein each R₅ is independently -NO₂, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, -NH₂, mono- or di-(Ci-C₆ alkyl) amino, or -C(O)ORg. Even more preferred are compounds of formula XIII, Z is quinolinyl optionally substituted with one or two R5 groups, wherein each R5 is independently -NO₂, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, -NH₂, mono- or di-(Ci-C₆ alkyl) amino, or -C(O)ORg.

[0129] In another preferred embodiment of compounds of formula XIII, R₂ is -H, -NO₂, -OH, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, mono- to per-halogenated Ci-C₆ alkyl, -NH₂, mono- or di-(Ci-C₆ alkyl) amino, -C(O)-OR₉, -N(R₈)-Z, -0-Z, -S-Z, -SO₂-Z, or -N(R₈)-Z. More preferred are compounds of formula XIII, wherein R₂ is -H, -NO₂, -OH, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, mono- to per-halogenated Ci-C₆ alkyl, -NH₂, mono- or di-(Ci-C₆ alkyl) amino, -C(O)-OR₉. Even more preferred are compounds of formula XIII, wherein R₂ is -H. [0130] In another preferred embodiment of compounds of formula XIII, R₃ is methyl. More preferred compounds of formula XIII are those where R3 is methyl and n is O, 1 , or 2. [0131] In a particular embodiment, the compounds of formula XIII are compounds of formula XIV, XV, XVI, XVII, or XVIII:

XIV XV

XVI XVII

XVIII.

wherein m is O, 1, 2, 3, or 4. [0132] In other preferred embodiments of the compounds of formula XIV, XV, or XVI, n is 0, 1, or 2. More preferably, n is 0. In other more preferred embodiments, n is 2. [0133] In other preferred embodiments of the compounds of formula XVIII, m is 0, 1 , or 2. More preferably, m is 1.

[0134] In another preferred embodiment of compounds of formula XV, XVI, or XVII, Y is aryl or heteroaryl wherein the aryl and heteroaryl is optionally substituted with one to four R₅ groups, wherein each R₅ is independently -NO₂, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, -NH₂, mono- or di-(Ci-C₆ alkyl) amino, or -C(O)OR₉.

[0135] Even more preferred are compounds of formula XV, XVI, OR XVII, wherein Y is aryl, optionally substituted with one or two R5 groups, wherein each R5 is independently -NO₂, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, -NH₂, mono- or (Ii-(C₁-C₆ alkyl) amino, or -C(O)OR₉. [0136] Other preferred compounds of formula XV, XVI, OR XVII are those wherein Y is heteroaryl, optionally substituted with one or two R₅ groups, wherein each R₅ is independently -NO₂, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, -NH₂, mono- or di-(Ci-C₆ alkyl) amino, or -C(O)OR₉.

[0137] Even more preferred are compounds of formula XV, XVI, OR XVII, wherein Y is phenyl, optionally substituted with one or two R₅ groups, wherein each R₅ is independently -NO₂, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, -NH₂, mono- or di-(Ci-C₆ alkyl) amino, or -C(O)OR₉.

[0138] Other even more preferred are compounds of formula XV, XVI, OR XVII, wherein Y is pyridyl, optionally substituted with one or two R₅ groups, wherein each R₅ is independently -NO₂, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, -NH₂, mono- or di-(Ci-C₆ alkyl) amino, or -C(O)OR₉.

[0139] In another preferred embodiment of compounds of formula XIV, XV, XVI, XVII, or XVIII, wherein Z is aryl or heteroaryl (preferably, quinolinyl) wherein the aryl and heteroaryl is optionally substituted with one to four R₅ groups, wherein each R₅ is independently -NO₂, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, -NH₂, mono- or di-(Ci-C₆ alkyl) amino, or -C(O)OR₉. More preferred are compounds of formula XIV, XV, XVI, XVII, or XVIII, wherein Z is aryl optionally substituted with one or two R₅ groups, wherein each R₅ is independently -NO₂, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, -NH₂, mono- or (Ii-(Ci-C₆ alkyl) amino, or -C(O)ORg. Even more preferred are compounds of formula XIV, XV, XVI, XVII, or XVIII, wherein Z is phenyl optionally substituted with one or two R5 groups, wherein each R5 is independently -NO₂, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, -NH₂, mono- or di-(Ci-C₆ alkyl) amino, or -C(O)ORg. [0140] In another preferred embodiment of compounds of formula XIV, XV, XVI, XVII, or XVIII, Z is heteroaryl (preferably, quinolinyl) optionally substituted with one or two R₅ groups, wherein each R₅ is independently -NO₂, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, -NH₂, mono- or di-(Ci-C₆ alkyl) amino, or -C(O)ORg. Even more preferred are compounds of formula XIV, XV,

XVI, XVII, or XVIII, Z is pyridyl optionally substituted with one or two R5 groups, wherein each R₅ is independently -NO₂, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, -NH₂, mono- or di-(Ci-C₆ alkyl) amino, or -C(O)ORg. Even more preferred are compounds of formula XIV, XV, XVI,

XVII, or XVIII, Z is quinolinyl optionally substituted with one or two R₅ groups, wherein each R₅ is independently -NO₂, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, -NH₂, mono- or di-(Ci-C₆ alkyl) amino, or -C(O)OR₉.

[0141] In another preferred embodiment of compounds of formula XIV, XV, XVI, XVII, or

XVIII, R₂ is -H, -NO₂, -OH, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, mono- to per-halogenated Ci-C₆ alkyl, -NH₂, mono- or (Ii-(C₁-C₆ alkyl) amino, -C(O)-OR₉, -N(Rg)-Z, -0-Z, -S-Z, -SO₂-Z, or -N(Rs)-Z. More preferred are compounds of formula XIV, XV, XVI, XVII, or XVIII, wherein R₂ is -H, -NO₂, -OH, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, mono- to per-halogenated Ci-C₆ alkyl, -NH₂, mono- or di-(Ci-C₆ alkyl) amino, -C(O)-OR₉. Even more preferred are compounds of formula XIV, XV, XVI, XVII, or XVIII, wherein R₂ is -H.

[0142] In another preferred embodiment of compounds of formula XIV, XV, or XVI, R3 is methyl. More preferred compounds of formula XIV, XV, or XVI are those where R₃ is methyl and n is O, 1, or 2.

[0143] In a particular embodiment, the compounds of formula XV are compounds of formula XIX or XX:

XIX XX. wherein p is 0, 1, 2, 3, or 4; and each R₆ is independently -NO₂, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, -NH₂, mono- or di-(Ci-C₆ alkyl) amino, or -C(O)OR₉.

[0144] In other preferred embodiments of the compounds of formula XIX or XX, n is 0, 1, or

2. More preferably, n is 0. In other more preferred embodiments, n is 2.

[0145] In other preferred embodiments of the compounds of formula XIX or XX, p is 0, 1, or

2. More preferably, p is 0 or 1. In other more preferred embodiments, p is 1.

[0146] In another preferred embodiment of compounds of formula XIX or XX, each R₆ is independently -NO₂, -CN, halo, Ci-C₆ alkyl, or Ci-C₆ alkoxy.

[0147] In another preferred embodiment of compounds of formula XIX or XX, each R₆ is independently -NO₂, -CN, halo, Ci-C₆ alkyl, or Ci-C₆ alkoxy, and p is 1 or 2.

[0148] In another preferred embodiment of compounds of formula XIX or XX, R₆ is -NO₂,

-CN, halo, Ci-C₆ alkyl, or Ci-C₆ alkoxy, and p is 1.

[0149] In another preferred embodiment of compounds of formula XIX or XX, wherein Z is aryl or heteroaryl (preferably, quinolinyl) wherein the aryl and heteroaryl is optionally substituted with one to four R₅ groups, wherein each R₅ is independently -NO₂, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, -NH₂, mono- or di-(Ci-C₆ alkyl) amino, or -C(O)OR₉. More preferred are compounds of formula XIX or XX, wherein Z is aryl optionally substituted with one or two R₅ groups, wherein each R₅ is independently -NO₂, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, -NH₂, mono- or di-(Ci-C₆ alkyl) amino, or -C(O)OR₉. Even more preferred are compounds of formula

XIX or XX, wherein Z is phenyl optionally substituted with one or two R₅ groups, wherein each

R₅ is independently -NO₂, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, -NH₂, mono- or di-(Ci-C₆ alkyl) amino, or -C(O)OR₉.

[0150] In another preferred embodiment of compounds of formula XIX or XX, Z is heteroaryl (preferably, quinolinyl) optionally substituted with one or two R₅ groups, wherein each R₅ is independently -NO₂, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, -NH₂, mono- or di-(Ci-C₆ alkyl) amino, or -C(O)OR₉. Even more preferred are compounds of formula XIX or XX, Z is pyridyl optionally substituted with one or two R₅ groups, wherein each R₅ is independently

-C(O)OR₉. Even more preferred are compounds of formula XIX or XX, Z is quinolinyl optionally substituted with one or two R₅ groups, wherein each R₅ is independently -NO₂, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, -NH₂, mono- or di-(Ci-C₆ alkyl) amino, or -C(O)OR₉. [0151] In another preferred embodiment of compounds of formula XIX or XX, R₂ is -H,

-NO₂, -OH, -CN, halo, C₁-C₆ alkyl, C₁-C₆ alkoxy, mono- to per-halogenated C₁-C₆ alkyl, -NH₂, mono- or (Ii-(C₁-C₆ alkyl) amino, -C(O)-OR₉, -N(Rg)-Z, -0-Z, -S-Z, -SO₂-Z, or -N(Rs)-Z. More preferred are compounds of formula XIX or XX, wherein R₂ is -H, -NO₂, -OH, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, mono- to per-halogenated Ci-C₆ alkyl, -NH₂, mono- or (Ii-(C₁-C₆ alkyl) amino, -C(O)-OR₉. Even more preferred are compounds of formula XIX or XX, wherein R₂ is

-H.

[0152] In another preferred embodiment of compounds of formula XIX or XX, R₃ is methyl.

More preferred compounds of formula XIX or XX, are those where R3 is methyl and n is O, 1, or

2.

[0153] In a particular embodiment, the compounds of formula XVI are compounds of formula XXI:

XXI

[0154] In another preferred embodiment of compounds of formula XXI, Y is aryl or heteroaryl wherein the aryl and heteroaryl is optionally substituted with one to four R₅ groups, wherein each R₅ is independently -NO₂, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, -NH₂, mono- or (Ii-(C₁-C₆ alkyl) amino, or -C(O)OR₉.

[0155] Even more preferred are compounds of formula XXI, wherein [0156] Y is aryl, optionally substituted with one or two R5 groups, wherein each R5 is independently -NO₂, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, -NH₂, mono- or CIi-(C₁-C₆ alkyl) amino, or -C(O)OR₉.

[0157] Other preferred compounds of formula XXI are those wherein Y is heteroaryl, optionally substituted with one or two R₅ groups, wherein each R₅ is independently -NO₂, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, -NH₂, mono- or (Ii-(C₁-C₆ alkyl) amino, or -C(O)OR₉. [0158] Even more preferred are compounds of formula XXI, wherein Y is phenyl, optionally substituted with one or two R₅ groups, wherein each R₅ is independently -NO₂, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, -NH₂, mono- or (Ii-(C₁-C₆ alkyl) amino, or -C(O)OR₉. [0159] Other even more preferred are compounds of formula XXI, wherein Y is pyridyl, optionally substituted with one or two R₅ groups, wherein each R₅ is independently -NO₂, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, -NH₂, mono- or di-(Ci-C₆ alkyl) amino, or -C(O)ORg. [0160] In another preferred embodiment of compounds of formula XXI, wherein Z is aryl or heteroaryl (preferably, quinolinyl) wherein the aryl and heteroaryl is optionally substituted with one to four R₅ groups, wherein each R₅ is independently -NO₂, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, -NH₂, mono- or di-(Ci-C₆ alkyl) amino, or -C(O)ORg. More preferred are compounds of formula XXI, wherein Z is aryl optionally substituted with one or two R₅ groups, wherein each R₅ is independently -NO₂, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, -NH₂, mono- or di-(Ci-C₆ alkyl) amino, or -C(O)ORg. Even more preferred are compounds of formula XXI, wherein Z is phenyl optionally substituted with one or two R₅ groups, wherein each R₅ is independently -NO₂, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, -NH₂, mono- or di-(Ci-C₆ alkyl) amino, or -C(O)ORg. [0161] In another preferred embodiment of compounds of formula XXI, Z is heteroaryl (preferably, quinolinyl)optionally substituted with one or two R₅ groups, wherein each R₅ is independently -NO₂, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, -NH₂, mono- or di-(Ci-C₆ alkyl) amino, or -C(O)ORg. Even more preferred are compounds of formula XXI, Z is pyridyl optionally substituted with one or two R₅ groups, wherein each R₅ is independently -NO₂, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, -NH₂, mono- or di-(Ci-C₆ alkyl) amino, or -C(O)OR₉. Even more preferred are compounds of formula XXI, Z is quinolinyl optionally substituted with one or two R₅ groups, wherein each R₅ is independently -NO₂, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, -NH₂, mono- or di-(Ci-C₆ alkyl) amino, or -C(O)ORg.

[0162] In another preferred embodiment of compounds of formula XXI, R₂ is -H, -NO₂, -OH, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, mono- to per-halogenated Ci-C₆ alkyl, -NH₂, mono- or (Ii-(C₁-C₆ alkyl) amino, -C(O)-OR₉, -N(Rg)-Z, -0-Z, -S-Z, -SO₂-Z, or -N(Rs)-Z. More preferred are compounds of formula XXI, wherein R₂ is -H, -NO₂, -OH, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, mono- to per-halogenated Ci-C₆ alkyl, -NH₂, mono- or di-(Ci-C₆ alkyl) amino, -C(O)-ORg. Even more preferred are compounds of formula XXI, wherein R₂ is -H. [0163] In another preferred embodiment of compounds of formula XXI, R₃ is methyl. [0164] In a particular embodiment, the compounds of formula XVII are compounds of formula XXII:

XXII wherein q is 0, 1, 2, 3, or 4; and each R₇ is independently -NO₂, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, -NH₂, mono- or di-(Ci-C₆ alkyl) amino, or -C(O)ORg.

[0165] In other preferred embodiments of the compounds of formula XXII, q is 0, 1, or 2.

More preferably, q is 0 or 1. In other more preferred embodiments,q is 1.

[0166] In another preferred embodiment of compounds of formula XXII, each R₇ is independently -NO₂, -CN, halo, Ci-C₆ alkyl, or Ci-C₆ alkoxy.

[0167] In another preferred embodiment of compounds of formula XXII, each R₇ is independently -NO₂, -CN, halo, Ci-C₆ alkyl, or Ci-C₆ alkoxy, and q is 1 or 2.

[0168] In another preferred embodiment of compounds of formula XXII, R₇ is -NO₂, -CN, halo, Ci-C₆ alkyl, or Ci-C₆ alkoxy, and q is 1.

[0169] In another preferred embodiment of compounds of formula XXII, wherein Z is aryl or heteroaryl (preferably, quinolinyl) wherein the aryl and heteroaryl is optionally substituted with one to four R₅ groups, wherein each R₅ is independently -NO₂, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, -NH₂, mono- or di-(Ci-C₆ alkyl) amino, or -C(O)OR₉. More preferred are compounds of formula XXII, wherein Z is aryl optionally substituted with one or two R₅ groups, wherein each

R₅ is independently -NO₂, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, -NH₂, mono- or di-(Ci-C₆ alkyl) amino, or -C(O)OR₉. Even more preferred are compounds of formula XXII, wherein Z is phenyl optionally substituted with one or two R₅ groups, wherein each R₅ is independently -NO₂, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, -NH₂, mono- or di-(Ci-C₆ alkyl) amino, or -C(O)OR₉.

[0170] In another preferred embodiment of compounds of formula XXII, Z is heteroaryl

(preferably, quinolinyl) optionally substituted with one or two R₅ groups, wherein each R₅ is independently -NO₂, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, -NH₂, mono- or (Ii-(C₁-C₆ alkyl) amino, or --C(O)OR₉. Even more preferred are compounds of formula XXII, Z is quinolinyl optionally substituted with one or two R₅ groups, wherein each R₅ is independently -NO₂, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, -NH₂, mono- or di-(Ci-C₆ alkyl) amino, or -C(O)OR₉. Even more preferred are compounds of formula XXII, Z is pyridyl optionally substituted with one or two R5 groups, wherein each R₅ is independently -NO₂, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy,

-NH₂, mono- or di-(Ci-C₆ alkyl) amino, or -C(O)OR₉.

[0171] In another preferred embodiment of compounds of formula XXII, R₂ is -H, -NO₂,

-OH, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, mono- to per-halogenated Ci-C₆ alkyl, -NH₂, mono- or (Ii-(C₁-C₆ alkyl) amino, -C(O)-OR₉, -N(R₈)-Z, -0-Z, -S-Z, -SO₂-Z, or -N(R₈)-Z. More preferred are compounds of formula XXII, wherein R₂ is -H, -NO₂, -OH, -CN, halo, Ci-C₆ alkyl,

Ci-C₆ alkoxy, mono- to per-halogenated Ci-C₆ alkyl, -NH₂, mono- or di-(Ci-C₆ alkyl) amino,

-C(O)-OR₉. Even more preferred are compounds of formula XXII, wherein R₂ is -H.

[0172] In a more preferred embodiment of the sixth aspect, the invention provides one of the compounds listed in the following table,

1 ■

[0173] In a seventh aspect, the invention provides a compound of formula (XL),

(XL) wherein, R is Ci-C₆ alkyl, Ci-C₆ haloalkyl, aryl, or heteroaryl, wherein R is optionally substituted with one to four R³ groups;

Li is L₃, -Ci-C₆ alkyl-, -Ci-C₆ alkyl-L₃-, -L₃-Ci-C₆ alkyl-, or -L₃-Ci-C₆ alkyl-L₃, wherein each L₃ is independently -C(O)N(R₄)-, -N(R₄)C(O)-, -OC(O)-, -C(O)O-, -N(R₄)C(O)N(R₄)-, -OC(O)N(R₄)-, -N(R₄)C(O)O-, -OC(O)O-, -C(O)-, -S(O)-, -S(O)₂-, -S(O)₂N(R₄)-, or -N(R₄)S(O)₂-; Ri is aryl or heteroaryl, wherein Ri is optionally substituted with one to four R groups; L₂ is -C(O)N(R₄)-, -N(R₄)C(O)-, -OC(O)-, -C(O)O-, -N(R₄)C(O)N(R₄)-, -OC(O)N(R₄)-, -N(R₄)C(O)O-, -OC(O)O-, -C(O)-, -S(O)-, -S(O)₂-, -S(O)₂N(R₄)-, or -N(R₄)S(O)₂-;

R₂ is aryl or heteroaryl, wherein R₂ is optionally substituted with one to four R₃ groups; and

R₃ is halo, -OR₆, -N(Re)₂, -S(R₆), -S(O)₂R₆, -S(O)₂N(Re)₂, -S(O)₂OR₆,

-N(R₆)S(O)₂R₆₅-OS(O)₂R₆, -C(O)R₆, -C(O)OR₆, -C(O)N(Re)₂, -OC(O)R₆, -OC(O)OR₆, -OC(O)N(Re)₂, -N(R₆)C(O)R₆, -N(R₆)C(O)OR⁶, -N(R₆)C(O)N(Re)₂, -CN, -NO₂, -Ci-C₆ alkyl, -C₁-C₆ haloalkyl, -Ci-C₆ alkyl-OR₆, or -Ci-C₆ alkyl-N(R₆)₂, wherein each R₆ is independently-H, Ci-C₆ alkyl, Ci-C₆ haloalkyl, C₃-Cg cycloalkyl, heterocyclyl, aryl, or heteroaryl; or any two R₃ on adjacent carbon atoms, taken together with the atoms to which they are attached, form a fused C₃-Cs cycloalkyl, heterocyclyl, aryl, or heteroaryl group; each R₄ is independently -H, Ci-C₆ alkyl, or Ci-C₆ haloalkyl, provided the compound is not

[0174] In a preferred embodiment of the seventh aspect, the compound of formula (XL)\ is of formula (XLI),

[0175] In preferred embodiments of the seventh aspect, the invention provides the compound of formula (XL) or (XLI), wherein R is Ci-C₆ alkyl. In other preferred embodiments of the seventh aspect, the invention provides the compound of formula (XL) or (XLI), wherein R is phenyl optionally substituted with one to four R₃ groups.

[0176] In another embodiment, the invention provides the compound of formula (XL) or (XLI), wherein Li is -Ci-C₆ alkyl-L₃- or -L₃-Ci-C₆ alkyl-, wherein each L₃ is independently -C(O)N(R₄)-, -N(R₄)C(O)-, -OC(O)-, -C(O)O-, -N(R₄)C(O)N(R₄)-, -OC(O)N(R₄)-, -N(R₄)C(O)O-, -OC(O)O-, -C(O)-, -S(O)-, -S(O)₂-, -S(O)₂N(R₄)-, or -N(R₄)S(O)₂-. In a more preferred embodiment, the invention provides the compound of formula (XL) or (XLI), wherein Li is -Ci-C₆ alkyl-L₃- or -L₃-Ci-C₆ alkyl-, wherein each L₃ is independently -C(O)N(R₄)-, -N(R₄)C(O)-, -S(O)₂N(R₄)-, or -N(R₄)S(O)₂-. In an even more preferred embodiment, the invention provides the compound of formula (XL) or (XLI), wherein Li is -Ci-C₆ alkyl-L₃- or -L₃-Ci-C₆ alkyl-, wherein each L₃ is independently -C(O)N(R₄)- or -N(R₄)C(O)-. [0177] In another embodiment, the invention provides the compound of formula (XL) or (XLI), wherein L₂ is -C(O)N(R₄)-, -N(R₄)C(O)-, -OC(O)-, -C(O)O-, -N(R₄)C(O)N(R₄)-, -OC(O)N(R₄)-, -N(R₄)C(O)O-, -OC(O)O-. In a more preferred embodiment, the invention provides the compound of formula (XL) or (XLI), wherein L₂ is -N(R₄)C(O)N(R₄)-, -OC(O)N(R₄)-, -N(R₄)C(O)O-, -OC(O)O-. In an even more preferred embodiment, the invention provides the compound of formula (XL) or (XLI), wherein L₂ is -N(R₄)C(O)N(R₄)-. [0178] In another embodiment, the invention provides the compound of formula (XL) or (XLI), wherein each R₃ is independently halo, -OR₆, -N(Re)₂, -S(O)₂R₆, -S(O)₂N(Rδ)₂, -C(O)R₆, -C(O)OR₆, -C(O)N(Re)₂, -CN, -C₁-C₆ alkyl, -C₁-C₆ haloalkyl, wherein each R₆ is independently-H, Ci-C₆ alkyl or Ci-C₆ haloalkyl. [0179] In a eighth aspect, the invention provides a compound of formula (LX),

(LX) wherein m and n are independently 0, 1, 2, 3, 4, or 5;

Ri and R₂ are independently -H, -Ci-C₆ alkyl, -aryl, -heteroaryl, -Ci-C₆ alkyl-aryl, or -Ci-C₆ alkyl-heteroaryl, wherein each is optionally substituted with one to four R₅ groups;

R₃, R₄, and R₅ are each independently halo, -OR₆, -N(Re)₂, -S(R₆), -S(O)₂R₆, -S(O)₂N(R6)₂, -S(O)₂OR₆, -N(R₆)S(O)₂R₆, -OS(O)₂R₆, -C(O)R₆, -C(O)OR₆, -C(O)N(Re)₂, -OC(O)R₆, -OC(O)OR₆, -OC(O)N(Rg)₂, -N(R₆)C(O)R₆, -N(R₆)C(O)OR⁶, -N(R₆)C(O)N(Rg)₂, -CN, -NO₂, -Ci-C₆ alkyl, -C₁-C₆ haloalkyl, -Ci-C₆ alkyl-ORg, or -Ci-C₆ alkyl-N(R₆)₂, wherein each R₆ is independently-H, Ci-C₆ alkyl, Ci-C₆ haloalkyl, C₃-Cg cycloalkyl, heterocyclyl, aryl, or heteroaryl, provided the compound is not

[0180] In preferred embodiments of the eighth aspect, the invention provides the compound of formula (LX), wherein m and n are independently 0, 1, or 2. More preferably, m and n are each 1.

[0181] In another embodiment, the invention provides the compound of formula (LX), wherein Ri and R₂ are independently -Ci-C₆ alkyl-aryl or -Ci-C₆ alkyl-heteroaryl, wherein each is optionally substituted with one to four R5 groups. More preferably, Ri and R₂ are independently -Ci-C₆ alkyl-aryl or -Ci-C₆ alkyl-heteroaryl, wherein each is optionally substituted with one to four groups which are independently halo, -ORs, -N(Rs)₂, -S(Rs), -S(O)₂Rs, -S(O)²N(Rs)₂, -C(O)R₈, -C(O)OR₈, -C(O)N(Rs)₂, -CN, -NO₂, -Ci-C₆ alkyl, or - Ci-C₆ haloalkyl, wherein each R₈ is independently -H, Ci-C₆ alkyl.

[0182] The compounds according to the sixth, seventh, and eighth aspects of the invention are also useful as general ubiquitin ligase inhibitors. For example, the compounds of the invention can be used as inhibitors of enzymes that exhibit ligase activity, including but not limited to p27/SCF, TRAF6, Radl8, BMI-I, APC and E3 enzymes. The compounds of the invention are also useful for regulating or inhibiting pathways in diseases and conditions that involve ubiquitin conjugation and deconjugation such as cancers, inflammation, metabolism, viral diseases and central nervous system disorders. For example, the compounds of the invention can be used to regulate or inhibit the products of genes that encode ubiquitin or ubiquitin-like enzymes described in Wong et al. (DDT 8 (16), 746-754 (2003)), which is incorporated by reference in its entirety.

[0183] In the ninth aspect, the invention provides for pharmaceutical compositions comprising, together with a pharmaceutically acceptable carrier, diluent, or excipient, a compound of according to the sixth, seventh, or eighth aspect of the invention described above or a compound selected from (2-((4-hydroxybenzyl)(4-methoxybenzyl)amino)-4-phenylthiazol-5- yl)(phenyl)methanone;

(2-((4-hydroxybenzyl)(4-methoxyphenethyl)amino)-4-(4-methoxyphenyl)thiazol-5-yl)(4- methoxyphenyl)methanone;

(2-((4-hydroxybenzyl)(4-methoxybenzyl)amino)-4-(4-methoxyphenyl)thiazol-5-yl)(4- methoxyphenyl)methanone;

(2-(butyl(4-hydroxybenzyl)amino)-4-phenylthiazol-5-yl)(phenyl)methanone; or

(2-amino-4-phenylthiazol-5-yl)(phenyl)methanone,

N-(benzo[d][l,3]dioxol-5-ylmethyl)-3-(tert-butylamino)-2-(4-(3-(4- (trifluoromethoxy)phenyl)ureido)phenyl)imidazo [ 1 ,2-a]pyridine-6-carboxamide; and

3-(2,6-dimethylphenylamino)-N-(4-methoxyphenethyl)-2-(2-(3-(4- methoxyphenyl)ureido)phenyl)imidazo[l,2-a]pyridine-8-carboxamide.

[0184] In the tenth aspect, the invention provides methods of inhibiting ubiquitination in a cell comprising contacting the cell in which inhibition of ubiquitination is desired with a compound according to the sixth, seventh, or eighth aspect of the invention or a pharmaceutical composition according to the ninth aspect of the invention. The compounds and formulations of the invention can inhibit ubiquitination in cells derived from animals, particularly, mammalian cells. The compounds and formulations of the invention can also be used to inhibit the ubiquitin ligase activity of p27/SCF, TRAF6, Radl8, or BMI-I.

[0185] In the eleventh aspect, the invention provides for methods of treating cell proliferative diseases or conditions comprising administering to a patient an effective amount of a compound of the sixth, seventh, or eighth aspect of the invention or a pharmaceutical composition according to the ninth aspect of the invention. Cell proliferative diseases or conditions include, but are not limited to, psoriasis, keloid scarring, and cancers, such as cancers of the breast, immune system, bone, nervous system, brain, blood, lymphatic system, and skin. Particularly, the compounds and pharmaceutical compositions of the invention are useful for treating cell proliferative diseases or conditions that involve p27/SCF, TRAF6, Radl8, or BMI-I.

[0186] In the twelfth aspect, the invention provides for methods of inhibiting TRAF6 comprising administering to a patient an effective amount of a compound according to the sixth, seventh, or eighth aspect of the invention or pharmaceutical composition according to the ninth aspect of the invention. Particularly, the compounds and pharmaceutical compositions are useful for treating conditions or diseases that involve TRAF6 such as those related to cancer, inflammation, adaptive immunity, innate immunity, bone metabolism, LPS-induced angiogenesis, osteoporosis, osteopinneal diseases, lymph node development, mammary gland development, skin development, and central nervous system development. [0187] In the thirteenth aspect, the invention provides for methods of inhibiting p27/SCF comprising administering to a patient an effective amount of a compound according to the sixth, seventh, or eighth aspect of the invention or pharmaceutical composition according to the ninth aspect of the invention. Particularly, the compounds and pharmaceutical compositions are useful for treating conditions or diseases that involve p27/SCF such as those related to cancer, inflammation, adaptive immunity, innate immunity, bone metabolism, LPS-induced angiogenesis, osteoporosis, osteopinneal diseases, lymph node development, mammary gland development, skin development, and central nervous system development. [0188] A variety of cellular proliferative disorders may be treated using the drug and prodrug compounds via the disclosed methods. In some embodiments, the drug or prodrug compounds are used to treat various cancers in afflicted subjects. Cancers are traditionally classified based on the tissue and cell type from which the cancer cells originate. Carcinomas are considered cancers arising from epithelial cells while sarcomas are considered cancers arising from connective tissues or muscle. Other cancer types include leukemias, which arise from hematopoietic cells, and cancers of nervous system cells, which arise from neural tissue. For non-invasive tumors, adenomas are considered benign epithelial tumors with glandular organization while chondomas are benign tumor arising from cartilage. In the present invention, the described compounds may be used to treat proliferative disorders encompassed by carcinomas, sarcomas, leukemias, neural cell tumors, and non-invasive tumors. [0189] Solid tumor cancers include malignant neoplastic masses of tissue or cancerous neoplasms characterized by the progressive or uncontrolled proliferation of cells. The cells involved in the neoplastic growth have an intrinsic heritable abnormality such that they are not regulated properly by normal methods. Malignant or cancerous neoplasms tend to grow rapidly, spread throughout the body, and recur if removed. The cells of malignant tumors may be well differentiated, but most have some degree of anaplasia. Anaplastic cells tend to be larger than normal and are abnormal, even bizarre, in shape. The nuclei tend to be very large, and irregular, and they often stain darkly. Malignant tumors may be partially encapsulated, but the cells of the cancer can infiltrate and destroy surrounding tissue. Thus, cells from the primary tumor can migrate (metastasize) from the original tumor site and colonize in other tissues. Tumors formed from cells that have spread are referred to as "secondary tumors" and contain cells that are similar to those in the original "primary" tumor. Metastatic tumors typically form by migration of tumor cells from the original tumor site through the blood and lymph system to other tissues. [0190] Specific properties of cancers, such as tissue invasiveness or metastasis, may be targeted using the methods described herein. In some embodiments, the drugs or prodrugs are used to treat solid tumors arising from various tissue types, including, but not limited to, cancers of the bone, breast, respiratory tract (e.g., bladder), brain reproductive organs, digestive tract, urinary tract, eye, liver, skin, head, neck, thyroid, parathyroid, and metastatic forms thereof. [0191] Specific proliferative disorders include the following: a) proliferative disorders of the breast include, but are not limited to, invasive ductal carcinoma, invasive lobular carcinoma, ductal carcinoma, lobular carcinoma in situ, and metastatic breast cancer; b) proliferative disorders of the skin include, but are not limited to, basal cell carcinoma, squamous cell carcinoma, malignant melanoma, and Karposi's sarcoma; c) proliferative disorders of the respiratory tract include, but are not limited to, small cell and non-small cell lung carcinoma, bronchial adema, pleuropulmonary blastoma, and malignant mesothelioma; d) proliferative disorders of the brain include, but are not limited to, brain stem and hyptothalamic glioma, cerebellar and cerebral astrocytoma, medullablastoma, ependymal tumors, oligodendroglial, meningiomas, and neuroectodermal and pineal tumors; e) proliferative disorders of the male reproductive organs include, but are not limited to, prostate cancer, testicular cancer, and penile cancer f) proliferative disorders of the female reproductive organs include, but are not limited to, uterine cancer (endometrial), cervical, ovarian, vaginal, vulval cancers, uterine sarcoma, ovarian germ cell tumor; g) proliferative disorders of the digestive tract include, but are not limited to, anal, colon, colorectal, esophageal, gallbladder, stomach (gastric), pancreatic cancer, pancreatic cancer- Islet cell, rectal, small-intestine, and salivary gland cancers; h) proliferative disorders of the liver include, but are not limited to, hepatocellular carcinoma, cholangiocarcinoma, mixed hepatocellular cholangiocarcinoma, and primary liver cancer; i) proliferative disorders of the eye include, but are not limited to, intraocular melanoma, retinoblastoma, and rhabdomyosarcoma; j) proliferative disorders of the head and cancers include, but are not limited to, laryngeal, hypopharyngeal, nasopharyngeal, oropharyngeal cancers, and lip and oral cancer, squamous neck cancer, metastatic paranasal sinus cancer; k) proliferative disorders of the lymphomas include, but are not limited to, various T cell and B cell lymphomas, non-Hodgkins lymphoma, cutaneous T cell lymphoma, Hodgkins disease, and lymphoma of the central nervous system; 1) leukemias include, but are not limited to, acute myeloid leukemia, acute lymphoblastic leukemia, chronic lymphocytic leukemia, chronic myelogenous leukemia, and hair cell leukemia, m) proliferative disorders of the thyroid include thyroid cancer, thymoma, and malignant thymoma; n) proliferative disorders of the urinary tract include, but are not limited to, bladder cancer; o) sarcomas include, but are not limited to, sarcoma of the soft tissue, osteosarcoma, malignant fibrous histiocytoma, lymphosarcoma, and rhabdomyosarcoma.

[0192] It is to be understood that the descriptions of proliferative disorders is not limited to the conditions described above, but encompasses other disorders characterized by uncontrolled growth and malignancy. It is further understood that proliferative disorders include various metastatic forms of the tumor and cancer types described herein. The drug and prodrug compounds of the described methods may be tested for effectiveness against these disorders, and a therapeutically effective regimen established. Effectiveness, as further described below, includes reduction or remission of the tumor, decreases in the rate of cell proliferation, or cytostatic or cytotoxic effect on cell growth.

[0193] In the fourteenth aspect, the invention provides for methods of inhibiting Radl8 comprising administering to a patient an effective amount of a compound according to the sixth, seventh, or eighth aspect of the invention or pharmaceutical composition according to the ninth aspect of the invention. Particularly, the compounds and pharmaceutical compositions are useful for treating conditions or diseases that involve Radl8 such as those related to cancer, inflammation, adaptive immunity, innate immunity, bone metabolism, LPS-induced angiogenesis, osteoporosis, osteopinneal diseases, lymph node development, mammary gland development, skin development, and central nervous system development. [0194] In the fifteenth aspect, the invention provides for methods of inhibiting BMI-I comprising administering to a patient an effective amount of a compound according to the sixth, seventh, or eighth aspect of the invention or pharmaceutical composition according to the ninth aspect of the invention. Particularly, the compounds and pharmaceutical compositions are useful for treating conditions or diseases that involve BMI-I such as those related to cancer, inflammation, adaptive immunity, innate immunity, bone metabolism, LPS-induced angiogenesis, osteoporosis, osteopinneal diseases, lymph node development, mammary gland development, skin development, and central nervous system development. [0195] The compounds for use in the method according to the twelfth aspect of the invention are also useful as general ubiquitin ligase inhibitors. For example, the compounds of the invention can be used as inhibitors of E3 enzymes that contain HECT and RING finger domains, Mdm2 with RING fingers and variants, and U-box-containing proteins. Accordingly, the compounds of the invention are useful as protein modulators, immunologic agents anti-inflammatory agents, anti-osteoporosis agents, anti-viral agents, for example, inhibitors of variola viruses such as smallpox, HIV and related conditions, human papillomavirus, HSV, adenovirus, coxsackie virus, HCMV, KSHV, EBV, paramyxovirus, myxomavirus, ebola, retrovirus, and rhabdovirus, anti-protozoan agents, for example, inhibitors of the malaria parasite. The compounds of the invention are also useful as oncologic and anti-pro liferative agents that inhibit aberrant cell growth, cancers, restenosis, psoriasis, and neoplastic cell proliferation.

[0196] In a sixteenth aspect, the invention provides a pharmaceutical composition comprising a pharmaceutically acceptable carrier, diluent, or excipient and a compound of formula (XXX),

(XXX) wherein, n is 0, 1, 2, or 3;

R¹ is -CN or -C(O)NH(R⁵), wherein R⁵ is -H, Ci-C₆ alkyl, or Ci-C₆ haloalkyl; R² is -H or -L-R⁷, wherein

L is -C(O)- or -S(O)₂-; and

R⁷ is Ci-C₆ alkyl, Ci-C₆ haloalkyl, C₃-C₈ cycloalkyl, heterocyclyl, aryl, or heteroaryl, wherein R⁷ is optionally substituted with one to four R⁴ groups; R is -Ci-C₆ alkyl, -Ci-C₆ haloalkyl, aryl, -Ci-C₆ alkyl-aryl, wherein R is optionally substituted with one to four R⁴ groups; and R⁴ is halo, -OR⁶, -N(R⁶)₂, -S(R⁶), -S(O)₂R⁶, -S(O)₂N(R⁶)₂, -S(O)₂OR⁶,

-N(R⁶)S(O)₂R⁶,-OS(O)₂R⁶, -C(O)R⁶, -C(O)OR⁶, -C(O)N(R⁶)₂, -OC(O)R⁶, -OC(O)OR⁶, -OC(O)N(R⁶)₂, -N(R⁶)C(O)R⁶, -N(R⁶)C(O)OR⁶, -N(R⁶)C(O)N(R⁶)², -CN, -NO₂, -Ci-C₆ alkyl, -Ci-C₆ haloalkyl, -C₁-C₆ alkyl-OR⁶, or -C₁-C₆ alkyl-N(R⁶)₂, wherein each R⁶ is independently-H, Ci-C₆ alkyl, Ci-C₆ haloalkyl, C₃-Cs cycloalkyl, heterocyclyl, aryl, or heteroaryl.

[0197] In a preferred embodiment of the seventh aspect, the compound of formula (XXX) is of formula (XXXI),

(XXXI).

[0198] In a preferred embodiment of the seventh aspect, the compound of formula (XXXI) is of formula (XXXII),

(XXXII).

[0199] In a preferred embodiment of the seventh aspect, the compound of formula (XXXII) is of formula (XXXIII),

[0200] In another preferred embodiment of the seventh aspect, the invention provides the pharmaceutical composition wherein the compound is one of formulas (XXX) - (XXXIII), and R⁷ is aryl or heteroaryl, wherein R⁷ is optionally substituted with one to four R⁴ groups. More preferred embodiments include those where the compound is one of formulas (XXX) - (XXXIII), and R⁷ is phenyl optionally substituted with one to four R⁴ groups. Even more preferred embodiments include those where the compound is one of formulas (XXX) - (XXXIII), and R⁷ is phenyl optionally substituted with one or two R⁴ groups. Other preferred embodiments include those where the compound is one of formulas (XXX) - (XXXIII), and R⁷ is furanyl, thienyl, pyrrolyl, thiazolyl, oxazolyl, isoxazoyl, isothiazoyl, imidazoyl, pyrazoyl, or triazolyl, wherein each R⁷ is optionally substituted with one to four R⁴ groups. Other more preferred embodiments include those where the compound is one of formulas (XXX) - (XXXIII), and R⁷ is furanyl, thienyl, or pyrrolyl, wherein each R⁷ is optionally substituted with one or two R⁴ groups. Other more preferred embodiments include those where the compound is one of formulas (XXX) - (XXXIII), and R⁷ is furanyl, thienyl, or pyrrolyl, wherein each R⁷ is optionally substituted with one or two halo groups. [0201] In another preferred embodiment of the seventh aspect, the invention provides the pharmaceutical composition wherein the compound is one of formulas (XXX) - (XXXIII), and [0202] R³ is -Ci-C₆ alkyl, phenyl, or benzyl, wherein R³ is optionally substituted with one t< four R⁴ groups. More preferred embodiments include those where the compound is one of formulas (XXX) - (XXXIII), and R³ is -Ci-C₆ alkyl, phenyl, or benzyl, wherein R³ is optionally substituted with one or two R⁴ groups. Even more preferred embodiments include those where the compound is one of formulas (XXX) - (XXXIII), and R³ is -Ci-C₆ alkyl optionally substituted with one R⁴ group. Even more preferred embodiments include those where the compound is one of formulas (XXX) - (XXXIII), and R³ is benzyl optionally substituted with one R⁴ group.

[0203] In a more preferred embodiment of the seventh aspect, the invention provides the pharmaceutical composition wherein the compound is one listed in the following table,

-

[0204] In the seventeenth aspect, the invention provides methods of inhibiting ubiquitination in a cell comprising contacting the cell in which inhibition of ubiquitination is desired with a pharmaceutical composition according to the sixteenth aspect of the invention. The compounds and formulations of the invention can inhibit ubiquitination in cells derived from animals, particularly, mammalian cells. The compounds and formulations of the invention can also be used to inhibit the ubiquitin ligase activity of p27/SCF, TRAF6, Radl8, or BMI-I. [0205] In the eighteenth aspect, the invention provides for methods of treating cell proliferative diseases or conditions comprising administering to a patient an effective amount of a pharmaceutical composition according to the sixteenth aspect of the invention. Cell proliferative diseases or conditions include, but are not limited to, psoriasis, keloid scarring, and cancers, such as cancers of the breast, immune system, bone, nervous system, brain, blood, lymphatic system, and skin. Particularly, the compounds and pharmaceutical compositions of the invention are useful for treating cell proliferative diseases or conditions that involve p27/SCF, TRAF6, Radl8, or BMI-I.

[0206] In the nineteenth aspect, the invention provides for methods of inhibiting TRAF6 comprising administering to a patient an effective amount of a pharmaceutical composition according to the sixteenth aspect of the invention. Particularly, the compounds and pharmaceutical compositions are useful for treating conditions or diseases that involve TRAF6 such as those related to cancer, inflammation, adaptive immunity, innate immunity, bone metabolism, LPS-induced angiogenesis, osteoporosis, osteopinneal diseases, lymph node development, mammary gland development, skin development, and central nervous system development.

[0207] In the twentieth aspect, the invention provides for methods of inhibiting p27/SCF comprising administering to a patient an effective amount of a pharmaceutical composition according to the sixteenth aspect of the invention. Particularly, the compounds and pharmaceutical compositions are useful for treating conditions or diseases that involve p27/SCF such as those related to cancer, inflammation, adaptive immunity, innate immunity, bone metabolism, LPS-induced angiogenesis, osteoporosis, osteopinneal diseases, lymph node development, mammary gland development, skin development, and central nervous system development.

[0208] In the twenty-first aspect, the invention provides for methods of inhibiting Radl8 comprising administering to a patient an effective amount of a pharmaceutical composition according to the sixteenth aspect of the invention. Particularly, the compounds and pharmaceutical compositions are useful for treating conditions or diseases that involve Radl8 such as those related to cancer, inflammation, adaptive immunity, innate immunity, bone metabolism, LPS-induced angiogenesis, osteoporosis, osteopinneal diseases, lymph node development, mammary gland development, skin development, and central nervous system development. [0209] In the twenty-second aspect, the invention provides for methods of inhibiting BMI-I comprising administering to a patient an effective amount of a pharmaceutical composition according to the sixteenth aspect of the invention. Particularly, the compounds and pharmaceutical compositions are useful for treating conditions or diseases that involve BMI-I such as those related to cancer, inflammation, adaptive immunity, innate immunity, bone metabolism, LPS-induced angiogenesis, osteoporosis, osteopinneal diseases, lymph node development, mammary gland development, skin development, and central nervous system development.

[0210] The pharmacetical compositions for use in the method according to the nineteenth aspect of the invention are also useful as general ubiquitin ligase inhibitors. For example, the compounds of the invention can be used as inhibitors of E3 enzymes that contain HECT and RING finger domains, Mdm2 with RING fingers and variants, and U-box-containing proteins. Accordingly, the compounds of the invention are useful as protein modulators, immunologic agents anti-inflammatory agents, anti-osteoporosis agents, anti-viral agents, for example, inhibitors of variola viruses such as smallpox, HIV and related conditions, human papillomavirus, HSV, adenovirus, coxsackie virus, HCMV, KSHV, EBV, paramyxovirus, myxomavirus, ebola, retrovirus, and rhabdovirus, anti-protozoan agents, for example, inhibitors of the malaria parasite. The compounds of the invention are also useful as oncologic and anti-proliferative agents that inhibit aberrant cell growth, cancers, restenosis, psoriasis, and neoplastic cell proliferation.

[0211] The compounds in the table above can be prepared using art recognized methods. All of the compounds in this application were named using ChemDraw Ultra version 8.0 or 10.0, which is available through Cambridgesoft.com, 100 Cambridge Park Drive, Cambridge, MA 02140, or Namepro version 6.0, which is available from ACD Labs, 90 Adelaide Street West, Toronto, Ontario, M5H, 3V9, Canada, or were derived therefrom.

[0212] For simplicity, chemical moieties are defined and referred to throughout primarily as univalent chemical moieties (e.g., alkyl, aryl, etc.). Nevertheless, such terms are also used to convey corresponding multivalent moieties under the appropriate structural circumstances clear to those skilled in the art. For example, while an "alkyl" moiety generally refers to a monovalent group (e.g. CH3-CH2-), in certain circumstances a bivalent linking moiety can be "alkyl," in which case those skilled in the art will understand the alkyl to be a divalent group (e.g. , -CH₂-CH₂-), which is equivalent to the term "alkylene." (Similarly, in circumstances in which a divalent moiety is required and is stated as being "aryl," those skilled in the art will understand that the term "aryl" refers to the corresponding divalent moiety, arylene.) All atoms are understood to have their normal number of valences for bond formation (i.e., 4 for carbon, 3 for N, 2 for O, and 2, 4, or 6 for S, depending on the oxidation state of the S). On occasion a moiety may be defined, for example, as (A)_a-B-, wherein a is 0 or 1. In such instances, when a is 0 the moiety is B- and when a is 1 the moiety is A-B-.

[0213] The term "absent" as used herein means that the element is not present in the structure of the compound.

[0214] The term "alkyl" as employed herein refers to straight and branched chain aliphatic groups having from 1 to 12 carbon atoms, preferably 1-8 carbon atoms, and more preferably 1-6 carbon atoms, which is optionally substituted with one, two or three substituents. Preferred alkyl groups include, without limitation, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl, and hexyl. A "Co" alkyl (as in "Co-C₃_alkyl") is a covalent bond. [0215] The term "alkenyl" as used herein means an unsaturated straight or branched chain aliphatic group with one or more carbon-carbon double bonds, having from 2 to 12 carbon atoms, preferably 2-8 carbon atoms, and more preferably 2-6 carbon atoms, which is optionally substituted with one, two or three substituents. Preferred alkenyl groups include, without limitation, ethenyl, propenyl, butenyl, pentenyl, and hexenyl.

[0216] The term "alkynyl" as used herein means an unsaturated straight or branched chain aliphatic group with one or more carbon-carbon triple bonds, having from 2 to 12 carbon atoms, preferably 2-8 carbon atoms, and more preferably 2-6 carbon atoms, which is optionally substituted with one, two or three substituents. Preferred alkynyl groups include, without limitation, ethynyl, propynyl, butynyl, pentynyl, and hexynyl.

[0217] An "alkylene," "alkenylene," or "alkynylene" group is an alkyl, alkenyl, or alkynyl group, as defined hereinabove, that is positioned between and serves to connect two other chemical groups. Preferred alkylene groups include, without limitation, methylene, ethylene, propylene, and butylene. Preferred alkenylene groups include, without limitation, ethenylene, propenylene, and butenylene. Preferred alkynylene groups include, without limitation, ethynylene, propynylene, and butynylene. [0218] The term "cycloalkyl" as employed herein includes saturated and partially unsaturated cyclic hydrocarbon groups having 3 to 12 carbons, preferably 3 to 8 carbons, and more preferably 3 to 6 carbons, wherein the cycloalkyl group additionally is optionally substituted. Preferred cycloalkyl groups include, without limitation, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cycloheptyl, and cyclooctyl.

[0219] The term "heteroalkyl" refers to an alkyl group, as defined hereinabove, wherein one or more carbon atoms in the chain are replaced by a heteroatom selected from the group consisting of O, S, and N.

[0220] An "aryl" group is a Ce-Cu aromatic moiety comprising one to three aromatic rings, which is optionally substituted. Preferably, the aryl group is a C₆-Ci_O aryl group. Preferred aryl groups include, without limitation, phenyl, naphthyl, anthracenyl, and fluorenyl. An "aralkyl" or "arylalkyl" group comprises an aryl group covalently linked to an alkyl group, either of which may independently be optionally substituted or unsubstituted. Preferably, the aralkyl group is (Ci-C₆)alkyl(C₆-Cio)aryl, including, without limitation, benzyl, phenethyl, and naphthylmethyl. Further, the term "bis-aryl" group comprises an aryl group covalently linked to an aryl or heteroaryl group, any one of which may independently be optionally substituted or unsubstituted. Preferably, the bis-aryl group is (C6-Cio)aryl(C6-Cio)aryl or (C6-C₁₀)aryl(C6-C₁₀)heteroaryl, including, without limitation, 2,1,3-benzoxadiazolyl substituted in the benzo portion with phenyl or quinolinyl.

[0221] A "heterocyclic" group (or "heterocyclyl") is an optionally substituted non-aromatic mono-, bi-, or tricyclic structure having from about 3 to about 14 atoms, wherein one or more atoms are selected from the group consisting of N, O, and S. One ring of a bicyclic heterocycle or two rings of a tricyclic heterocycle may be aromatic, as in indan and 9,10-dihydro anthracene. The heterocyclic group is optionally substituted on carbon with oxo or with one of the substituents listed above. The heterocyclic group is also optionally independently be substituted on nitrogen with alkyl, aryl, aralkyl, alkylcarbonyl, alkylsulfonyl, arylcarbonyl, arylsulfonyl, alkoxycarbonyl, aralkoxycarbonyl, or on sulfur with oxo or lower alkyl. Preferred heterocyclic groups include, without limitation, epoxy, aziridinyl, tetrahydrofuranyl, pyrrolidinyl, piperidinyl, piperazinyl, thiazolidinyl, oxazolidinyl, oxazolidinonyl, and morpholino. In certain preferred embodiments, the heterocyclic group is fused to an aryl, heteroaryl, or cycloalkyl group. Examples of such fused heterocycles include, without limitation, tetrahydroquinoline and dihydrobenzofuran. Specifically excluded from the scope of this term are compounds an annular O or S atom is adjacent to another O or S atom.

[0222] As used herein, the term "heteroaryl" refers to optionally substituted groups having 5 to 14 ring atoms, preferably 5, 6, 9, or 10 ring atoms; having 6, 10, or 14 pi electrons shared in a cyclic array; and having, in addition to carbon atoms, between one or more heteroatoms selected from the group consisting of N, O, and S. For example, a heteroaryl group may be pyrimidinyl, pyridinyl, benzimidazolyl, thienyl, benzothiazolyl, benzofuranyl and indolinyl. Preferred heteroaryl groups include, without limitation, thienyl, benzothienyl, furyl, benzofuryl, dibenzofuryl, pyrrolyl, imidazolyl, pyrazolyl, pyridyl, pyrazinyl, pyrimidinyl, indolyl, quinolyl, isoquinolyl, quinoxalinyl, tetrazolyl, oxazolyl, thiazolyl, and isoxazolyl.

[0223] A "heteroaralkyl" or "heteroarylalkyl" group comprises a heteroaryl group covalently linked to an alkyl group, either of which is independently optionally substituted or unsubstituted. Preferred heteroalkyl groups comprise a Ci-C₆ alkyl group and a heteroaryl group having 5, 6, 9, or 10 ring atoms. Specifically excluded from the scope of this term are compounds having adjacent annular O and/or S atoms. Examples of preferred heteroaralkyl groups include pyridylmethyl, pyridylethyl, pyrrolylmethyl, pyrrolylethyl, imidazolylmethyl, imidazolylethyl, thiazolylmethyl, and thiazolylethyl.

[0224] An "arylene," "heteroarylene," or "heterocyclylene" group is an aryl, heteroaryl, or heterocyclyl group, as defined hereinabove, that is positioned between and serves to connect two other chemical groups.

[0225] Preferred heterocyclyls and heteroaryls include, but are not limited to, acridinyl, azocinyl, benzimidazolyl, benzofuranyl, benzothiofuranyl, benzothiophenyl, benzoxazolyl, benzthiazolyl, benztriazolyl, benztetrazolyl, benzisoxazolyl, benzisothiazolyl, benzimidazolinyl, carbazolyl, 4aH-carbazolyl, carbolinyl, chromanyl, chromenyl, cinnolinyl, decahydroquinolinyl, 2H,6H-l,5,2-dithiazinyl, dihydrofuro[2,3-b]tetrahydrofuran, furanyl, furazanyl, imidazolidinyl, imidazolinyl, imidazolyl, lH-indazolyl, indolenyl, indolinyl, indolizinyl, indolyl, 3H-indolyl, isobenzofuranyl, isochromanyl, isoindazolyl, isoindolinyl, isoindolyl, isoquinolinyl, isothiazolyl, isoxazolyl, methylenedioxyphenyl, morpholinyl, naphthyridinyl, octahydroisoquinolinyl, oxadiazolyl, 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,5-oxadiazolyl, 1,3,4-oxadiazolyl, oxazolidinyl, oxazolyl, oxazolidinyl, pyrimidinyl, phenanthridinyl, phenanthrolinyl, phenazinyl, phenothiazinyl, phenoxathiinyl, phenoxazinyl, phthalazinyl, piperazinyl, piperidinyl, piperidonyl, 4-piperidonyl, piperonyl, pteridinyl, purinyl, pyranyl, pyrazinyl, pyrazolidinyl, pyrazolinyl, pyrazolyl, pyridazinyl, pyridooxazole, pyridoimidazole, pyridothiazole, pyridinyl, pyridyl, pyrimidinyl, pyrrolidinyl, pyrrolinyl, 2H-pyrrolyl, pyrrolyl, quinazolinyl, quinolinyl, 4H-quinolizinyl, quinoxalinyl, quinuclidinyl, tetrahydrofuranyl, tetrahydroisoquinolinyl, tetrahydroquinolinyl, tetrazolyl, 6H-l,2,5-thiadiazinyl, 1,2,3-thiadiazolyl, 1,2,4-thiadiazolyl, 1,2,5-thiadiazolyl, 1,3,4-thiadiazolyl, thianthrenyl, thiazolyl, thienyl, thienothiazolyl, thienooxazolyl, thienoimidazolyl, thiophenyl, triazinyl, 1,2,3-triazolyl, 1,2,4-triazolyl, 1,2,5-triazolyl, 1,3,4-triazolyl, and xanthenyl.

[0226] As employed herein, when a moiety (e.g., cycloalkyl, aryl, heteroaryl, heterocyclic, urea, etc.) is described as "optionally substituted" it is meant that the group optionally has from one to four, preferably from one to three, more preferably one or two, non-hydrogen substituents. Suitable substituents include, without limitation, halo, hydroxy, oxo (e.g., an annular -CH- substituted with oxo is -C(O)-) nitro, alkyl, alkenyl, alkynyl, cycloalkyl, haloalkyl, haloalkenyl, haloalkynyl, halocycloalkyl, aryl, aralkyl, alkoxy, aryloxy, amino, acylamino, alkylcarbamoyl, arylcarbamoyl, aminoalkyl, acyl, carboxy, hydroxyalkyl, alkanesulfonyl, arenesulfonyl, alkanesulfonamido, arenesulfonamido, aralkylsulfonamido, alkylcarbonyl, acyloxy, cyano, and ureido groups. Preferred substituents, which are themselves not further substituted (unless expressly stated otherwise) are:

(a) halo, cyano, oxo, carboxy, formyl, nitro, amino, amidino, guanidino,

(b) C1-C5 alkyl or alkenyl or arylalkyl imino, carbamoyl, azido, carboxamido, mercapto, hydroxy, hydroxyalkyl, alkylaryl, arylalkyl, Ci-Cs alkyl, Ci-Cs alkenyl, Ci-Cs alkoxy, C₁-C₈ alkoxycarbonyl, aryloxycarbonyl, C₂-Cs acyl, C₂-Cs acylamino, Ci-Cs alkylthio, arylalkylthio, arylthio, Ci-Cs alkylsulfmyl, arylalkylsulfmyl, arylsulfmyl, Ci-Cs alkylsulfonyl, arylalkylsulfonyl, arylsulfonyl, Co-C₆ N-alkyl carbamoyl, C₂-C 15 JV,iV-dialkylcarbamoyl, C3-C7 cycloalkyl, aroyl, aryloxy, arylalkyl ether, aryl, aryl fused to a cycloalkyl or heterocycle or another aryl ring, C3-C7 heterocycle, or any of these rings fused or spiro-fused to a cycloalkyl, heterocyclyl, aryl, or heteroaryl, wherein each of the foregoing is further optionally substituted with one more moieties listed in (a), above; and

(c) -(CH₂)_s-N(R^3o)(R³¹), wherein s is from 0 (in which case the nitrogen is directly bonded to the moiety that is substituted) to 6, and R³° and R³¹ are each independently hydrogen, cyano, oxo, carboxamido, amidino, Ci-Cg hydroxyalkyl, C1-C3 alkylaryl, aryl-Ci-C₃ alkyl, Ci-C₈ alkyl, Ci-C₈ alkenyl, Ci-C₈ alkoxy, Ci-C₈ alkoxycarbonyl, aryloxycarbonyl, aryl-Ci-C₃ alkoxycarbonyl, C₂-C₈ acyl, Ci-C₈ alkylsulfonyl, arylalkylsulfonyl, arylsulfonyl, aroyl, aryl, cycloalkyl, heterocyclyl, or heteroaryl, wherein each of the foregoing is further optionally substituted with one more moieties listed in (a), above; or

R³° and R³¹ taken together with the N to which they are attached form a heterocyclyl or heteroaryl, each of which is optionally substituted with from 1 to 3 substituents from (a), above.

[0227] In addition, substituents on cyclic moieties (i.e., cycloalkyl, heterocyclyl, aryl, heteroaryl) include 5-6 membered mono- and 9-14 membered bi-cyclic moieties fused to the parent cyclic moiety to form a bi- or tri-cyclic fused ring system. For example, an optionally substituted phenyl includes, but not limited to, the following:

[0228] A "haloalkyl," "haloalkenyl," "haloalkynyl," or "halocycloalkyl" is an alkyl, alkenyl, alkynyl, or cycloalkyl moiety in which from one to all hydrogens have been replaced with one or more halo.

[0229] The term "halogen" or "halo" as employed herein refers to chlorine, bromine, fluorine, or iodine. As herein employed, the term "acyl" refers to an alkylcarbonyl or arylcarbonyl substituent. The term "acylamino" refers to an amide group attached at the nitrogen atom (i.e., R-CO-NH-). The term "carbamoyl" refers to an amide group attached at the carbonyl carbon atom (i.e., NH₂-CO-). The nitrogen atom of an acylamino or carbamoyl substituent is additionally substituted. The term "sulfonamido" refers to a sulfonamide substituent attached by either the sulfur or the nitrogen atom. The term "amino" is meant to include NH₂, alkylamino, arylamino, and cyclic amino groups. The term "ureido" as employed herein refers to a substituted or unsubstituted urea moiety.

[0230] A moiety that is substituted is one in which one or more hydrogens have been independently replaced with another chemical substituent. As a non-limiting example, substituted phenyls include 2-flurophenyl, 3,4-dichlorophenyl, 3-chloro-4-fluoro-phenyl, 2-fluoro-3-propylphenyl. As another non-limiting example, substituted iV-octyls include 2,4 dimethyl-5-ethyl-octyl and 3-cyclopentyl-octyl. Included within this definition are methylenes (-CH₂-) substituted with oxygen to form carbonyl -CO-).

[0231] An "unsubstituted" moiety as defined above (e.g., unsubstituted cycloalkyl, unsubstituted heteroaryl, etc.) means that moiety as defined above that does not have any of the optional substituents for which the definition of the moiety (above) otherwise provides. Thus, for example, while an "aryl" includes phenyl and phenyl substituted with a halo, "unsubstituted aryl" does not include phenyl substituted with a halo.

[0232] Throughout the specification preferred embodiments of one or more chemical substituents are identified. Also preferred are combinations of preferred embodiments. Any compounds excluded from the scope of a particular genus of compounds of the invention (e.g., through phrases beginning "provided that when...") are intended to be excluded from all other genera of compounds.

[0233] Some compounds of the invention may have chiral centers and/or geometric isomeric centers (E- and Z- isomers), and it is to be understood that the invention encompasses all such optical, diastereoisomers and geometric isomers. The invention also comprises all tautomeric forms of the compounds disclosed herein.

[0234] "Atropisomers" are stereoisomers resulting from hindered rotation about single bonds where the barrier to rotation is high enough to allow for the isolation of the conformers (Eliel, E. L.; Wilen, S. H. Stereochemistry of Organic Compounds; Wiley & Sons: New York, 1994; Chapter 14, including pages 1150-1153 and the short definition on page 1193). Atropisomerism is significant because it introduces an element of chirality in the absence of stereogenic atoms. The invention is meant to encompass atropisomers, for example in cases of limited rotation around the single bonds emanating from the core 2,1,3-benzoxadiazole or 2,1,3-benzothiadiazole structure. Atropisomers are also possible and are also specifically included in the compounds and/or prodrugs of the invention.

[0235] Polymorphism in chemical substances is the ability of a single compound to exist in two or more solid phases, each having different arrangements and/or conformations of the individual molecules in the solid form (D. J. W. Grant, Theory and Origin of Polymorphism. In H. G. Brittain (ed.) Polymorphism in Pharmaceutical Solids. Marcel Dekker, Inc., New York, 1999, pp. 1-34). Generally, polymorphic solid forms can be crystalline or amorphous. Polymorphs of molecules or their solvates (for example, hydrates) can exist. Distinct polymorphic forms generally have different chemical and physical properties such as melting point, chemical reactivity, apparent solubility, dissolution rate, optical and electrical properties, vapor pressure, and density. The invention is meant to encompass in its scope, different polymorphic forms of the compounds of the invention.

[0236] The compounds of the invention may be administered in the form of an in vivo hydrolyzable ester or in vivo hydrolyzable amide. An in vivo hydrolyzable ester of a compound of the invention containing carboxy or hydroxy group is, for example, a pharmaceutically acceptable ester which is hydrolyzed in the human or animal body to produce the parent acid or alcohol. Suitable pharmaceutically acceptable esters for carboxy include Ci_6-alkoxymethyl esters {e.g., methoxymethyl), Ci_₆-alkanoyloxymethyl esters {e.g., for example pivaloyloxymethyl), phthalidyl esters, C₃_₈-cycloalkoxycarbonyloxyCi_₆-alkyl esters {e.g., 1-cyclohexylcarbonyloxyethyl); l,3-dioxolen-2-onylmethyl esters {e.g., 5-methyl-l,3-dioxolen-2-onylmethyl; and Ci_₆-alkoxycarbonyloxyethyl esters {e.g., 1-methoxycarbonyloxy ethyl) and may be formed at any carboxy group in the compounds of this invention.

[0237] An in vivo hydrolyzable ester of a compound of the invention containing a hydroxy group includes inorganic esters such as phosphate esters and a-acyloxyalkyl ethers and related compounds which as a result of the in vivo hydrolysis of the ester breakdown to give the parent hydroxy group. Examples of α-acyloxyalkyl ethers include acetoxymethoxy and 2,2-dimethylpropionyloxy-methoxy. A selection of in vivo hydrolyzable ester forming groups for hydroxy include alkanoyl, benzoyl, phenylacetyl and substituted benzoyl and phenylacetyl, alkoxycarbonyl (to give alkyl carbonate esters), dialkylcarbamoyl and

TV^TVTV-dialkylaminoethy^-TV-alkylcarbamoyl (to give carbamates), TV,TV-dialkylaminoacetyl and carboxyacetyl. Examples of substituents on benzoyl include morpholino and piperazino linked from a ring nitrogen atom via a methylene group to the 3- or 4- position of the benzoyl ring. A suitable value for an in vivo hydrolyzable amide of a compound of the invention containing a carboxy group is, for example, a TV-Ci_6-alkyl or N,N-di-Ci_6-alkyl amide such as TV-methyl, TV-ethyl, TV-propyl, TVTV-dimethyl, TV-ethyl-TV-methyl or TVTV-diethyl amide.

Pharmaceutical Compositions

[0238] In a second aspect, the invention provides pharmaceutical compositions comprising an inhibitor of histone deacetylase according to the invention and a pharmaceutically acceptable carrier, excipient, or diluent. Compounds of the invention may be formulated by any method well known in the art and may be prepared for administration by any route, including, without limitation, parenteral, oral, sublingual, transdermal, topical, intranasal, intratracheal, or intrarectal. In certain preferred embodiments, compounds of the invention are administered intravenously in a hospital setting. In certain other preferred embodiments, administration may preferably be by the oral route.

[0239] The characteristics of the carrier will depend on the route of administration. As used herein, the term "pharmaceutically acceptable" means a non-toxic material that is compatible with a biological system such as a cell, cell culture, tissue, or organism, and that does not interfere with the effectiveness of the biological activity of the active ingredient(s). Thus, compositions according to the invention may contain, in addition to the inhibitor, diluents, fillers, salts, buffers, stabilizers, solubilizers, and other materials well known in the art. The preparation of pharmaceutically acceptable formulations is described in, e.g., Remington's The Science and Practice of Pharmacy, 20th Edition, 2000.

[0240] As used herein, the term pharmaceutically acceptable salts refers to salts that retain the desired biological activity of the above-identified compounds and exhibit minimal or no undesired toxicological effects. Examples of such salts include, but are not limited to acid addition salts formed with inorganic acids (for example, hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, nitric acid, and the like), and salts formed with organic acids such as acetic acid, oxalic acid, tartaric acid, succinic acid, malic acid, ascorbic acid, benzoic acid, tannic acid, pamoic acid, alginic acid, polyglutamic acid, naphthalenesulfonic acid, naphthalenedisulfonic acid, and polygalacturonic acid. The compounds can also be administered as pharmaceutically acceptable quaternary salts known by those skilled in the art, which specifically include the quaternary ammonium salt of the formula -NR + W-, wherein R is R₃-R₇, and W is a counterion, including chloride, bromide, iodide, -O-alkyl, toluenesulfonate, methylsulfonate, sulfonate, phosphate, or carboxylate (such as benzoate, succinate, acetate, glycolate, maleate, malate, citrate, tartrate, ascorbate, benzoate, cinnamoate, mandeloate, benzyloate, and diphenylacetate). As used herein, the term "salt" is also meant to encompass complexes, such as with an alkaline metal or an alkaline earth metal.

[0241] The active compound is included in the pharmaceutically acceptable carrier or diluent in an amount sufficient to deliver to a patient a therapeutically effective amount without causing serious toxic effects in the patient treated. A preferred dose of the active compound for all of the above-mentioned conditions is in the range from about 0.01 to 300 mg/kg, preferably 0.1 to 100 mg/kg per day, more preferably 0.1 to about 50 mg per kilogram body weight of the recipient per day, and in some applications about 0.1 to about 25 mg per kilogram body weight of the recipient per day. A typical topical dosage will range from 0.01-3% wt/wt in a suitable carrier. The effective dosage range of the pharmaceutically acceptable derivatives can be calculated based on the weight of the parent compound to be delivered. If the derivative exhibits activity in itself, the effective dosage can be estimated as above using the weight of the derivative, or by other means known to those skilled in the art.

Synthetic Schemes and Experimental Procedures

[0242] The compounds of the invention may be synthesized according to the methods known to those of ordinary skill in the art. For example, methods that may be used to make the compounds of the invention are described in Mallory, F. B. (Organic Syntheses, Coll. Vol. IV: pp 74-75 (John Wiley & Sons, 1963)); Smith, P.A.S. and Boyer, J.H. (Organic Syntheses, Coll. Vol. IV: pp 75-78 (John Wiley & Sons, 1963)), and in Can. J. Chem., pp 2482-2484 (1969). These references are incorporated by references in their entirety.

[0243] Scheme I outlines the general approach taken to synthesize the 2,1,3-benzoxadiazole scaffold. There are also commercially available 2,1,3-benzoxadiazoles and 2,1,3-benzothiadiazoles that can be used as starting materials for compounds of the invention. Schemes H-V outline some more specific synthetic methods used to make particular compounds of the invention.

Scheme I Synthesis of 2,li3-Benzoxadiazoles

"benzofurazan oxide" 2,1 ,3-benzoxadiazole

"benzofurazan oxide"

2,1 ,3-benzoxadiazole

[0244] Referring to Scheme I, the 2,1,3-benzoxadiazole scaffold is prepared starting with a 1 ,2-dinitrogen substituted aryl system. In the first two reaction paths in Scheme I, a 1,2-bis aniline is used, for example. In the first reaction path, the 1,2-bis aniline is treated with NaOCl and NaOH in a ring closure reaction to give the corresponding benzofurazan oxide, which in turn is reduced to the corresponding 2,1,3-benzoxadiazole. Alternatively, the 1,2-bis aniline can be treated with NaNO₂ZHCl (Sandmeyer reaction chemistry) to make an intermediate diazonium salt, which when treated with sodium azide (NaN₃) gives the corresponding ortho-amino phenyl azide. Heating the aryl azide in toluene gives the corresponding benzofurazan oxide, which, as outlined above, is reduced to the corresponding 2,1,3-benzoxadiazole. The final reaction path in Scheme I shows that an ortho-amino nitrobenzene can be converted to the corresponding 2,1,3-benzoxadiazole by first converting the amino group to a carbamate and then heating to high temperature.

Scheme II

[0245] Scheme II outlines how compounds of the invention having a 4-sulfonamide group are made generally. A 4-amino-2,l,3-benzoxadiazole is treated with a desired sulfonyl chloride in the presence of an acid scavenger, preferably an amine base. Scheme II shows two preferred methods, one using pyridine as the base and heating to 95 ⁰C to effect the reaction, and the other using a resin-bound morpholine as the base and heating to 45 ⁰C to effect the reaction. Specific examples are described below and provide detailed synthetic procedures.

Scheme IV

[0246] Scheme IV outlines how compounds of the invention having a 7-nitrogen-4-sulfur substitution are made generally. As depicted in the first reaction path, the appropriate 4-choro-7-nitro-2,l,3-benzoxadiazole is reacted with for example a sulfide to give the corresponding 7-nitrogen-4-sulfur containing analog. The sulfur group can be further oxidized to the corresponding sulfoxide or sulfone to make additional compounds of the invention. Alternatively, as depicted in the second reaction path, compounds of the invention having a substituted 7-chloro-4-sulfur substitution are made via reaction of the appropriate benzoxadiazole sulfonyl chloride with, for example, an amine partner to form sulfonamides and the like. In the example shown, the 7-choro group on the product (with sulfonamide installed) can further be exchanged via substitution reactions with thiols and amines for example (as shown in the specific examples) to make the corresponding 4-sulfur-7-sulfur-containing-2, 1 ,3-benzoxadiazoles and 4-sulfur-7-nitrogen-containing-2, 1 ,3-benzoxadiazoles, respectively. Scheme V

[0247] Scheme V outlines how compounds of the invention having, for example, a bis-aryl substitution are made generally. As depicted, the appropriate choro -2,1,3-benzoxadiazole is reacted with, for example, an aryl boronic acid under palladium-catalysis conditions to give the corresponding bis-aryl analog.

Scheme VI

12

[0248] Scheme VI outlines how compounds of the invention having sulfonyl and a halo, thioaryl, or thioheteroaryl susbstituents on the benzoxadiazole core. Generally, 2,6-dichloroaniline can be oxidixzed with MCPBA to yield the corresponding nitroso compound. Reaction of the nitroso compound with sodium azide gives the

4-chlorobenzo[c][l,2,5]oxadiazole. Chlorosulfonation with chlorosulfonic acid produces 7-chlorobenzo[c][l,2,5]oxadiazole-4-sulfonyl chloride. The sulfonamide linkage is formed by further reaction with the appropriate nitrogen-containing heterocycle; in Scheme VI, a substituted piperidine was used to give l-(7-chlorobenzo[c][l,2,5]oxadiazol-4-ylsulfonyl)-4-(4-chlorophenyl)piperidin-4-ol. Finally, reaction with quinonline-8-thiol hydrochloride in the presence of a base, for example, KOH, gives the desired final product as illustrated in Scheme VI.

Scheme VII

[0249] Scheme VII outlines how compounds of the invention having sulfonyl and a Z group linked to the core through a 'Q' moiety. In Scheme VII, Q represents -O-, -S-, or -N(R8)-, wherein each variable is as defined for formula (XX).

Scheme VIII

808 [0250] Scheme VIII illustrates the synthesis of compounds of the invention of general formula (XL). First compounds 801 and 802 were reacted to give a Schiff base which is further reacted in the same pot with an isonitrile (803) to form compound 804 which contains the imidazo[l,2-a]pyridine core. Deprotection with LiOH in aqueous THF gave the carboxylic acid, 805. Compound 806 was formed by standard amide coupling chemistry, in this case, HBTU in DMF with DIPEA, to yield 806. The nitro group was reduced with 10% Pd/C in methanol to give 807, and finally, reaction of the free amine in 807 with a isocyanate produced the desired compound of the invention, 808.

Scheme IX

[0251] Scheme IX illustrates the synthesis of compounds of the invention of general formula (XXX). In Scheme IX, each variable is as defined for formula (XXX). Reaction of 2,3-dichloroquinoxaline, 901, with malononitrile in the presence of a base gives 902. Next, reaction of 902 with a primary amine, followed by an acidic workup gives compound of the form of 903 with a lH-pyrrolo[2,3-b]quinoxaline core. Finally, reaction of 903 with a substituted acid chloride yields compounds of the invention, 904.

Scheme X

[0252] Scheme X illustrates the synthesis of compounds of the invention of general formula (LX) one a solid support. Similar methods for preparation of compounds of the invention can be found in Katritzky, J. Comb. Chem. 2003, 392, which is hereby incorporated by reference. [0253] Schemes I - X in conjunction with the examples described below, will make it sufficiently clear to one of ordinary skill in the art how to make the compounds of the invention. Compounds in Table 1 were made using the techniques described herein and were isolated and characterized by either ¹H-NMR, LC/MS or both. Commercially available starting materials, for example, amines, thiols, aryl halides, sulfonyl chlorides, boronic acids, were used in most cases along with the chemistry described to make the compounds of the invention. Example 1

8-[(7-Nitro-2,l,3-benzoxadiazol-4-yl)thio]quinoline (Cpd No. 5)

[0254] To 40 mg (0.2 mmol) of 4-chloro-7-nitro-2,l,3-benzoxadiazole was added 2 ml of DMF and 60 mg of potassium carbonate. To this reaction mixture was added 128 mg (4 equivalents) of 8-mercaptoquinoline. The reaction turned bright red. The reaction mixture was heated at 60⁰C for 4 hrs. Examination of the reaction by TLC showed no starting material. The reaction mixture was pored into 100 ml of water and extracted with ethyl acetate and methylene chloride. The combined organic layers were dried and the solvent evaporated. The residue was purified by column chromatography on silica gel using 1 : 2 EtOAc :Hexane as eluant. The 8-[(7-nitro-2,l,3-benzoxadiazol-4-yl)thio]quinoline was obtained as a red solid. 48.9 mg (75.4% yield)

[0255] ¹H NMR (CDCl₃): δ 6.45 (d, IH, J = 7.8 Hz); 7.55 (m,lH); 7.7 (dd, IH, J = 7.8 and 1.5 Hz); 8.1 (dd, 2 H, J = 1.5 and 7.8 Hz); 8.2 (dd, IH, J = 1.5 and 7.8 Hz); 8.3 (dd, IH, J = 1.5 and 7.8 Hz); 8.95 (dd, J = 1.5 and 7.8 Hz); LC/MS purity 100%. MS 325 (M+l) seen.

Example 2

4-(3,5-Dimethylphenyl)-2,l,3-benzoxadiazole (Cpd. no. 285)¹

[0256] A vial containing a suspension of 4-chloro-2,l,3-benzoxadiazole (77 mg, 0.5 mmol), 3,5-dimethylphenylboronic acid (105 mg, 0.7 mmol), potassium fluoride (87 mg, 1.5 mmol), palladium (II) acetate (3 mg, 0.005 mmol), and 2-(dicyclohexylphosphino)biphenyl (3.5 mg, 0.01 mmol) in toluene (3 mL) was twice degassed and back-filled with argon and capped. The reaction mixture was heated at 60 C for 15 hours, cooled, diluted with ethyl acetate, washed successively with IN NaOH and brine, and dried over MgSO₄. The organic layer was filtered through a plug of celite and silica gel and eluted with ethyl acetate. The filtrate was concentrated and the residue was purified by radial silica gel chromatography to afford Cpd No. 285 (54 mg, 48%).

[0257] ¹H NMR (CDCl₃, 300 MHz) δ 7.77 (d, J = 8.7 Hz, IH), 7.58 (br s, 2H), 7.55-7.42 (m, 2H), 7.10 (s, IH) 2.43 (s, 6H); LCMS purity: 100%. MS (positive ion): Found 225 (MH⁺). [0258] * Reference for procedure: J. Am. Chem. Soc. 1999, 121, 9550.

Example 3

4-[3-(Trifluoromethyl)phenyl]-2,l,3-benzoxadiazole (Cpd. No. 305)

[0259] This compound was prepared according to the procedure for the preparation of Cpd.

No. 285 and was purified by reverse phase liquid chromatography.

[0260] ¹H NMR (CDCl₃, 300 MHz) δ 8.24 (m, IH), 8.21 (m, IH), 7.87 (, d, J = 9 Hz, IH),

7,74-7.65 (m, 2H), 7.64 (d, J = 6.9 Hz, IH), 7.56-7.51 (m, IH).; MS (negative ion): Found 263

(M-H⁺).

Example 4

4-(3-Methylphenyl)-2,l,3-benzoxadiazole (Cpd. No. 306)

[0261] This compound was prepared according to the procedure for the preparation of Cpd. No. 285 and was purified by reverse phase liquid chromatography.

[0262] ¹H NMR (CDCl₃, 300 MHz) δ 7.80-7.76 (m, 3H), 7.57-7.46 (m, 2H), 7.41 (app t, J = 14.4 Hz, IH), 7.29-7.25 (m, IH), 2.48 (s, 3H); MS (negative ion): Found 209 (M-H⁺). Example 5

4-(4-Tert-butylphenyl)-2,l,3-benzoxadiazole (Cpd. No. 307)

[0263] This compound was prepared according to the procedure for the preparation of Cpd. No. 285 and was purified by reverse phase liquid chromatography.

[0264] ¹H NMR (CDCl₃, 300 MHz) δ 7.93 (d, J = 6.6 Hz, 2H), 7.78 (d, J = 8.7 Hz, IH), 7.57-7.46 (m, 4H), 1.40 (s, 9H); MS (negative ion): Found 251 (M-H⁺).

Example 6

4-(3,5-Dimethylphenyl)-7-nitro-2,l,3-benzoxadiazole (Cpd. No. 286)

[0265] This compound was prepared according to the procedure for the preparation of Cpd. No. 285 and was purified by reverse phase liquid chromatography.

[0266] ¹H NMR (CDCl₃, 300 MHz) δ 8.56 (d, J = 7.8 Hz, IH), 7.71 (d, J = 7.5 Hz, IH), 7.64 (s, 2H), 7.22 (m, IH), 2.46 (s, 6H); MS (negative ion): Found 269 (M).

Example 7

4-[(3,5-Difluoro)phenyl]-7-nitro-2,l,3-benzoxadiazole (Cpd. No. 308)

[0267] This compound was prepared according to the procedure for the preparation of Cpd. No. 285 and was purified by reverse phase liquid chromatography. [0268] ¹H NMR (CDCl₃, 300 MHz) δ 8.58 (d, J = 7.5 Hz, IH), 7.78 (d, J = 7.9 Hz, IH), 7.66-7.59 (m, 2H), 7.04 (tt, J = 2.1 8.7 Hz, IH); MS (negative ion): Found 277 (M).

Example 8

4-Nitro-7-[3-(trifluoromethyl)phenyl]-2,l,3-benzoxadiazole (Cpd. No. 309)

[0269] This compound was prepared according to the procedure for the preparation of Cpd. No. 285 and was purified by reverse phase liquid chromatography.

[0270] ¹H NMR (CDCl₃, 300 MHz) δ 8.61 (d, J = 7.8 Hz, IH), 8.28-8.26 (m, 2H), 7.86-7.72 (m, 3H); MS (negative ion): Found 309 (M).

Example 9

4-(3-Methylphenyl)-7-nitro-2,l,3-benzoxadiazole (Cpd. No. 310)

[0271] This compound was prepared according to the procedure for the preparation of Cpd. No. 285 and was purified by reverse phase liquid chromatography.

[0272] ¹H NMR (CDCl₃, 300 MHz) δ 8.58 (d, J = 7.8 Hz, IH), 7.85-7.83 (m, 2H), 7.73 (, d, J = 7.5 Hz, IH), 7.48 (app t, J = 7.8 Hz, IH), 7.40-7.38 (m, IH), 2.51 (s, 3H); MS (negative ion): Found 255 (M). Example 9

4-Nitro-7-[4-(trifluoromethyl)phenyl]-2,l,3-benzoxadiazole (Cpd. No. 311)

[0273] This compound was prepared according to the procedure for the preparation of Cpd. No. 285 and was purified by reverse phase liquid chromatography.

[0274] ¹H NMR (CDCl₃, 300 MHz) δ 8.61 (d, J = 7.5 Hz, IH), 8.16 (d, J = 8.7 Hz, IH), 7.87-7.80 (m, 3H); MS (negative ion): Found 309 (M).

Example 10

4-[2,4-Bis(trifluoromethyl)phenyl]-7-nitro-2,l,3-benzoxadiazole (Cpd. No. 312)

[0275] This compound was prepared according to the procedure for the preparation of Cpd. No. 285 and was purified by reverse phase liquid chromatography.

[0276] ¹H NMR (CDCl₃, 300 MHz) δ 8.58 (d, J = 7.5 Hz, IH), 8.15 (s, IH), 8.00 (d, J = 8.1 Hz, IH), 7.67 (d, J = 8.1 Hz, IH), 7.58 (d, J = 7.2 Hz, IH); MS (negative ion): Found 377 (M).

Example 11

4-(4-Tert-butylphenyl)-7-nitro-2,l,3-benzoxadiazole (Cpd. No. 313)

[0277] This compound was prepared according to the procedure for the preparation of Cpd. No. 285 and was purified by reverse phase liquid chromatography.

[0278] ¹H NMR (CDCl₃, 300 MHz) δ 8.58 (d, J = 7.8 Hz, IH), 8.01 (d, J = 8.4 Hz, 2H), 7.73 (d, J = 7.8 Hz, IH), 7.61 (d, J = 8.1 Hz, 2H), 1,41 (s, 9H); MS (negative ion): Found 297 (M).

Example 12

4-(2-Fluoro-3-methoxyphenyl)-7-nitro-2,l,3-benzoxadiazole (Cpd. No. 314)

[0279] This compound was prepared according to the procedure for the preparation of Cpd. No. 285 and was purified by reverse phase liquid chromatography.

[0280] ¹H NMR (CDCl₃, 300 MHz) δ 8.58 (d, J = 7.8 Hz, IH), 7.79 (d, J = 7.8 Hz, IH), 7.44-7.40 (m, IH), 7.28-7.25 (m, IH), 7.19-7.09 (m, IH), 3.98 (s, 3H).

Example 13

7-[4-(Methylthio)phenyl]-4-nitro-2,l,3-benzoxadiazole (Cpd No. 318)

[0281] This compound was prepared according to the procedure for the preparation of Cpd No. 285 and was purified by reverse phase liquid chromatography.

[0282] ¹H NMR (CDCl₃, 300 MHz) δ 8.57 (d, J = 7.8 Hz, IH), 8.03 (d, J = 8.4 Hz, 2H), 7.72 (d, J = 7.5 Hz, IH), 7.41 (d, J = 8.4 Hz, 2H), 2.58 (s, 3H); MS (positive ion): Found 288 (MH⁺). Example 14

7-(3-Methoxyphenyl)-4-nitro-2,l,3-benzoxadiazole (Cpd No. 319)

[0283] This compound was prepared according to the procedure for the preparation of Cpd No. 285 and was purified by reverse phase liquid chromatography.

[0284] ¹H NMR (CDCl₃, 300 MHz) δ 8.57 (d, J = 7.8 Hz, IH), 7.75 (d, J = 7.8 Hz, IH), 7.62-7.59 (m, 2H), 7.50 (t, J = 8.1 Hz, IH), 7.14-7.11 (m, IH), 3.92 (s, 3H); MS (positive ion): Found 272 (MH⁺).

Example 15

7-(3-Fluorophenyl)-4-nitro-2,l,3-benzoxadiazole (Cpd No. 320)

[0285] This compound was prepared according to the procedure for the preparation of Cpd No. 285 and was purified by reverse phase liquid chromatography.

[0286] ¹H NMR (CDCl₃, 300 MHz) δ 8.59 (d, J = 7.8 Hz, IH), 7.87-7.84 (m, IH), 7.88 (dt, J = 2.4, 9.6 Hz, IH), 7.77 (d, J = 7.5 Hz, IH), 7.61-7.54 (m, IH), 7.29 (app ddt, J = 0.6, 2.4, 7.5 Hz, IH); MS (positive ion): Found 260 (MH⁺).

Example 16

4-Nitro-7-(quinolin-8-yl)- 2,1,3-benzoxadiazole (Cpd No. 321) [0287] This compound was prepared according to the procedure for the preparation of Cpd No. 285 and was purified by reverse phase liquid chromatography.

[0288] ¹H NMR (CDCl₃, 300 MHz) δ 8.88 (app. dd, J = 1.2, 5.1 Hz, IH), 8.66 (d, J = 7.8 Hz, IH), app. dd, J = 1.2, 8.1 Hz, IH), 8.14 (d, J = 6.9 Hz, IH), 8.06 (t, J = 7.8 Hz, 2H), 7.75 (t, J = 8.1 Hz, IH), 7.52 (dd, J = 4.2, 8.7 Hz, IH); MS (positive ion): Found 293 (MH⁺).

Example 17

4-Chloro-6-[3,5-(dimethyl)phenyl]- 2,1,3-benzoxadiazole (Cpd No. 322) and

Example 18

4-Chloro-6-[3,5-(difluoro)phenyl]- 2,1,3-benzoxadiazole (Cpd No. 324)

[0289] This compound was prepared according to the procedure for the preparation of Cpd. No. 285 and was purified by reverse phase liquid chromatography.

[0290] ¹H NMR (DMSO-de, 300 MHz) δ 8.39 (t, J = 1.5 Hz, IH), 8.05 (t, J = 1.2 Hz, IH), 7.88-7.84 (m, 2H), 7.45 (dtt, J = 0.9, 2.1, 8.4 Hz, IH); MS (positive ion): Found 267 (MH⁺).

Example 19

4-Chloro-6-[3-(trifluoromethyl)phenyl]- 2,1,3-benzoxadiazole (Cpd No. 325)

[0291] This compound was prepared according to the procedure for the preparation of Cpd. No. 285 and was purified by reverse phase liquid chromatography. [0292] ¹H NMR (DMSO-de, 300 MHz) δ 8.40 (br s, IH), 8.38-8.35 (m, 2H), 8.05 (d, J = 1.2 Hz, IH), 7.90-7.87 (m, IH), 7.82-7.78 (m, IH); MS (positive ion): Found 299 (MH⁺).

Example 20

4-Chloro-6-(3-methylphenyl)- 2,1,3-benzoxadiazole (Cpd No. 326)

[0293] This compound was prepared according to the procedure for the preparation of Cpd. No. 285 and was purified by reverse phase liquid chromatography.

[0294] ¹H NMR (DMSO-de, 300 MHz) δ 8.27 (t, J = 1.8 Hz, IH), 7.86 (br s, IH), 7.83-7.82 (m, 2H), 7.44 (t, J = 7.8 Hz, IH), 7.33 (d, J = 7.8 Hz, IH), 2.41 (s, 3H); MS (positive ion): Found 245 (MH⁺).

Example 21

4-Chloro-6-[4-(trifluoromethyl)phenyl]- 2,1,3-benzoxadiazole (Cpd No. 327) [0295] This compound was prepared according to the procedure for the preparation of Cpd. No. 285 and was purified by reverse phase liquid chromatography. [0296] ¹H NMR (DMSO-d₆, 300 MHz) δ 8.39-8.38 (m, IH), 8.28 (d, J = 8.1 Hz, 2H), 8.01-8.00 (m, IH), 7.93 (d, J = 8.7 Hz, 2H); MS (positive ion): Found 299 (MH⁺).

Example 22

4-Chloro-6-(3-methoxyphenyl)- 2,1,3-benzoxadiazole (Cpd No. 328) [0297] This compound was prepared according to the procedure for the preparation of Cpd. 285 and was purified by reverse phase liquid chromatography.

[0298] ¹H NMR (DMSO-d₆, 300 MHz) δ 8.29 (m, IH), 7.89 (m, IH), 7.65-7.62 (m, IH), 7.59 (t, J = 2.4 Hz, IH), 7.47 (t, J = 7.8 Hz, IH), 7.09 (dd, J = 2.7, 8.1 Hz, IH), 3.84 (s, 3H); MS (positive ion): Found 261 (MH⁺).

Example 23

4-Chloro-6-(4-(tørt-butyl)phenyl)- 2,1,3-benzoxadiazole (Cpd No. 329)

[0299] This compound was prepared according to the procedure for the preparation of Cpd.

No. 285 and was purified by reverse phase liquid chromatography.

[0300] ¹H NMR (DMSO-de, 300 MHz) δ 8.24 (d, J = 1.5 Hz, IH), 7.98 (d, J = 8.1 Hz, 2H),

7.80 (d, J = 1.5 Hz, IH), 7.56 (d, J = 8.7 Hz, 2H), 1.33 (s, 9H); MS (positive ion): Found 287

(MH⁺).

Example 24

4-Chloro-6-(2-methoxyphenyl)- 2,1,3-benzoxadiazole (Cpd No. 330)

[0301] This compound was prepared according to the procedure for the preparation of Cpd. No. 285 and was purified by reverse phase liquid chromatography.

[0302] ¹H NMR (DMSO-de, 300 MHz) δ 8.28-8.27 (m, IH), 7.60-7.59 (m, IH), 7.57-7.55 (m, IH), 7.52-7.46 (m, IH), 7.21 (d, J = 8.7 Hz, IH), 7.09 (t, J = 7.5 Hz, IH), 3.76 (s, 3H); MS (negative ion): Found 259 (M-H⁺). Example 25

4-Chloro-6-(3-nitrophenyl)- 2,1,3-benzoxadiazole (Cpd No. 331)

[0303] This compound was prepared according to the procedure for the preparation of Cpd. No. 285 and was purified by reverse phase liquid chromatography. [0304] ¹H NMR (DMSO-d₆, 300 MHz) δ 8.92 (t, J = 2.1 Hz, IH), 8.53-8.50 (m, IH), 8.41-8.40 (m, IH), 8.38-8.35 (m, IH), 8.12-8.11 (m, IH), 7.87 (t, J = 8.1 Hz, IH); MS positive ion): Found 276 (MH⁺).

Example 34

Analysis of Compounds by LC/MS

[0305] The compounds of the invention were characterized by LC/MS using methods with various conditions. All the methods comprised a mobile phase that included 0.05% formic acid in water (component A) and 0.05% formic acid in acetonitrile (component B) for making the gradient. Table 3 below list the various conditions of the methods.

Table 3

Table 3

Biological Examples

[0306] For the assays described below, the TRAF6/UevlA /UBC 13 assay is the biochemical plate-based assay of TRAF 6 ligase activity using Ubcl3 as the E2 enzyme. The gel-based assay is the biochemical solution-based TRAF6 ligase assay (by SDS-PAGE and Western blot) to confirm the result from the plate-based ELISA assay. The APC assay is an assay which is another E3 ligase (APC2/APC11) biochemical assay.

Biological Example 1 TRAF6/UevlA/Ubcl3 Assay

[0307] A 96-well Ni-plate was blocked with 100 μl of 1% casein in PBS for 1 hour at room temperature. The plates were then washed three times with 200 μl of Ix PBS and 80 μl of a reaction buffer was added per well which contained 50 ng of Flag-ubiquitin, 50ng of His-ubiquitin, 62.5 mM Tris pH 7.5, 6.25 mM MgCl₂, 1.0 mM DTT, and 2 μM ATP. To each well was added 10 μl of a solution of the compound in DMSO. The reaction was started by adding 10 μl of a solution consisting of IOng human El, 25ng each of Uevla and Ubcl3, and lOOng TRAF6 in the reaction buffer. The plates were shaken for 10 minutes and incubate at room temperature for 1 hour. After the incubation, the plates were washed three times with 200 μl Ix PBS in 0.05% Tween and lOOμl of an antibody mix consisting of anti-Flag (1 :30,000 dilution; Sigma F-3165) and anti-Mouse IgG-HRP (1/150,000 dilution; Jackson Immunoresearch #115-035-146) in Ix PBS with 0.25% BSA. The plates were then incubated for another 1 hour at room temperature, and after incubation the plate was washed three times with 200 μl Ix PBS with 0.05% Tween. The amount of ubiquitin was measured by adding 100 μl of Lumino substrate (1 :5 dilution) and reading the luminescence with a fluorimeter. Table 2 contains data for this assay when example compounds of the invention were tested.

The following compounds of the invention were assayed as above. In the following table, IC₅O values are listed by range, where A = < 5 μM, B= 5-20 μM, and C = > 20 μM.

Biological Example 2 ICAM Assay

[0308] The expression of intercellular adhesion molecule (ICAM)-I in endothelial cells is pivotal in supporting lymphocyte migration across the vascular endothelium. TNF/IL-1 induces the NF-kB pathway in which NF -kB acts as a transcription factor and activation of NF-kB induces ICAM expression. IkB inhibits NF-kB by binding to it and retaining NF-kB in the cytoplasm. Upon cytokine stimulation, several signaling molecule including TRAF6, IKKs, and TAKl are activated which lead to the phosphorylation of IkB. Phosphorylated IkB is then ubiquitinated by the SCF complex and degraded by proteasome, thereby releasing NF-kB to translocate into the nucleus. In the nucleus, NF-kB binds to DNA and activates transcription of various genes that are involved in inflammation, cell survival, and apoptosis. The ICAM assay is a primary assay for HTS as well as a cell based assay for TRAF6 inhibitors.

[0309] The following materials are used 96-well plates; F12K complete medium that includes 10% FBS and 1% P/S (penicillin/streptomycin); IX PBS; 5 ug/ml IL-I, Human CD54

(CALTAG Laboratory, Cat#MHCD5400-4); anti-mouse-IgG (Jackson Lab, Cat#l 15-035 146); and A549 cells.

[0310] Procedure. 10,000 A549 cells/well were seeded in lOOμl F12K complete medium in a

96-well white plate. The seeded plate was incubate at 37⁰C incubator in 5% CO₂ overnight. After the overnight incubation, 4μl of diluted test compound was added and incubated for 1 hour at

37⁰C. After the 1 hour incubation, 4μl of 25ng/ml IL-I was added to each well and the cells were stimulated at 37⁰C for 4 hours .

[0311] After stimulation, the cells were stained with CD54 (1 :1000 diluted with medium) and anti-mouse IgG (1 : 1100 diluted with medium) for 1 hour. Subsequently, the medium was removed and each well was washed with 200μl of PBS three times. Detection of activity was performed by addition of lOOμl/well of lumino substrate.

Biological Example 3 Gel-Based E3 Ligase Assay

[0312] E3 (TRAF6) auto-ubiquitination was measured as described below. Activity in the presence of compound was determined relative to a parallel control in which only DMSO is added. The IC50 values were typically determined using 6 or 8 different concentrations of compound, although as few as 2 concentrations may be used to approximate the IC₅₀ values. [0313] E&K 96_well plates (E&K-20201) were used for the solution based biochemical assay. 80 μl of the reaction buffer were added to each well that contained 100 ng/well of Flag ubiquitin. To this, 10 μl of the test compound diluted in DMSO were added. After the test compound was added, 10 μl of El (human), E2 (Ubcl3/UevlA) and TRAF6 in Protein Buffer were added to obtain a final concentration of 10 ng/well of El, 25 ng/well of E2 and 100 ng/well TRAF6. The plates were shaken for 10 minutes and incubated at room temperature for 1 hour. After incubation, the reaction was stopped by adding 33μl of 4x loading buffer (non-reducing) per well and the plates were heated at 95°C for 5 minutes. An aliquot of each well was run on a 4-12% Bis-Tris NuPage Gel and analyzed by Western Blot using anti-Flag as primary antibody, and HRP- conjugated anti-Mouse IgG as secondary antibody. [0314] The Blocking Buffer contained 1% Casein in PBS. It was stored at 4°C until used.

[0315] The reaction buffer consisted of 62.5 mM Tris pH 7.6 (Trizma Base - Sigma T-8524),

3 mM MgCl₂ (Magnesium Chloride - Sigma M-2393), 1 mM DTT (Sigma D-9779), 2.5 mM

ATP (Roche Boehringer Mann Corp. 635-316), 100 ng/well of Flag-ubiquitin, 0.1% BSA (Sigma

A-7906), and 0.05% Tween-20 (Sigma P-7949).

[0316] The Protein Buffer consisted of 20 mM Tris pH 7.6, 10% glycerol (Sigma G-5516) and 1 mM DTT.

[0317] The antibody mix consisted of 0.25% BSA (Sigma A-7906) in IX PBS, 1/50,000 anti-Flag (Sigma F-3165), 1/100,000 of anti-Mouse IgG-HRP (Jackson Immunoresearch

#115-035-146).

[0318] The substrate mix consisted of SuperSignal Substrate from Pierce (catalog number

37070ZZ) and was prepared by mixing 100 ml of the peroxide solution, 100 ml of the enhancer solution and 100 ml of Milli-Q water.

[0319] The data from these gel experiments was confirmatory and in agreement with the

TRAF6/UB13 plate-based data.

Biological Example 4 APC-I l/APC-2 Ligase Assay

[0320] E3 (His APCl 1/APC2 - "APC") auto ubiquitination was measured as described in US Patent Application No. 09/826,312 (Publication No. US 2002 0042083 Al), which is incorporated by reference in its entirety. Details of the protocol are described below. Activity in the presence of the compound was determined relative to a parallel control in which only DMSO was added. Values of the IC50 were typically determined using 6 or 8 different concentrations of the compound, although as few as 2 concentrations may be used to approximate the IC50 value.

[0321] Nickel coated 96 well plates (Pierce 15242) were blocked for 1 hour with 100 μl of blocking buffer at room temperature. The plates were washed 4 times with 225 μl of 1 DPBS and 80 μl of the reaction buffer were added that contained 100 ng/well of Flag ubiquitin. To this, 10 μl of the test compound diluted in DMSO were added. After the test compound was added, 10 μl of El (human), E2 (Ubch5c), and APC in Protein Buffer was added to obtain a final concentration of 5 ng/well of El, 20 ng/well of E2 and 100 ng/well of APC. The plates were shaken for 10 minutes and incubated at room temperature for 1 hour. After incubation, the plates were washed 4 times with 225 μl of IxPBS and 100 μl/well of Antibody Mix were added to each well. The plates were incubated at room temperature for another hour after which they were washed 4 times with 225 μl of IxPBS and 100 μl/well of Lumino substrate were added to each well. The luminescence was measured by using a BMG luminescence microplate reader. The

Blocking Buffer (1% Casein in IxPBS) was stored at 4°C until use.

[0322] The reaction buffer consisted of 62.5 mM Tris pH 7.6 (Trizma Base - Sigma T 8524),

3 mM MgC12 (Magnesium Chloride - Sigma M 2393), 1 mM DTT (Sigma D 9779), 2.5 mM

ATP (Roche Boehringer Mann Corp. 635 316), 100 ng/well of Flag ubiquitin, 0.1% BSA (Sigma

A 7906), and 0.05% Tween 20 (Sigma P 7949).

[0323] The Protein Buffer consisted of 20 mM Tris pH 7.6, 10% glycerol (Sigma G 5516) and 1 mM DTT.

[0324] The antibody mix consisted of 0.25% BSA (Sigma A 7906) in IX PBS, 1/50,000 anti

Flag (Sigma F 3165), 1/100,000 of anti Mouse IgG HRP (Jackson Immunoresearch #115 035

146).

[0325] The substrate mix consisted of SuperSignal Substrate from Pierce (catalog number

37070ZZ) and was prepared by mixing 100 ml of the peroxide solution, 100 ml of the enhancer solution and 100 ml of Milli Q® water.

Biological Example 5

ROC1/CUL1 Ubiquitin Ligase Assay (SCF Assay)

[0326] Inhibition of ubiquitin ligase activityof E1+E2+E3 was measured using the protocol as described in WO 01/75145 with E3 as the ROC1/CUL1, ROC1/CUL2, or ROC2/CUL5 complex.

[0327] Materials and Methods

[0328] The wells of nickel-substrate 96-well plates (Pierce Chemical) are blocked with 100 μl of 1 casein/phosphate buffered saline (PBS) for 1 hour at room temperature, then washed with 200 μl of PBST (0. 1% Tween-20 in PBS) 3 times. To each well is added the following Flag-ubiquitin (see above) reaction solution: 62.5mM Tris pH 7.5, 6.25 mm MgC12, 0.75 mM DTT, 2.5 mM ATP, 2.5 mM NaFl, 2.5 nM Okadaic acid, 100 ng Flag-ubiquitin (made as described above).

[0329] The buffer solution is brought to a final volume of80 μl with Milipore-filtered water, followed by the addition of 10 μl DMSO. [0330] To the above solution is then added 10 μl of ubiquitination enzymes in 20 mM Tris buffer, pH 7.5, and 5% glycerol. E2-Ubch5c and E3-His ROCl/Cull, ROC1/CUL2, and ROC2/CUL5 are made as described in WO 01/75145. El is obtained commercially (Affϊniti Research Products, Exeter, U. K.). The following amounts of each enzyme are used for these assays: 5 ng/well of El; 25 nl/well E2; and 100 ng/well His-E3. Varying amounts of compounds according to the invention are added and the reaction allowed to proceed at room temperature for 1 hour.

[0331] Following the ubiquitination reaction, the wells are washed with 200 μl of PBST 3 times. For measurement of the enzyme-bound ubiquitin, 100 gel of Mouse anti-Flag (1 : 10,000) and anti-Mouse Ig-HRP (1 :15, 000) in PBST are added to each well and allowed to incubate at room temperature for 1 hour. The wells are then washed with 200 μl of PBST 3 times, followed by the addition of 100 μl of luminol substrate (1/5 dilution). Luminescence for each well is then measured using a fluorimeter.

Biological Example 6 Cell Proliferation Assays

Cell Culture Preparation

[0332] Cell lines used are available from American Type Culture Collection (ATCC), for example, cell cultures containing A549 (ATCC# CCL-185), HeLa (ATCC# CCL-2), HCTl 16 (ATCC# CCL-247), and H1299 (ATCC# CRL-5803) cells were maintained in Tl 75 flasks following the ATCC recommended media and handling procedures. Flasks reaching approximately 70% confluency were trypsinized and resuspended in RPMI media (Cell-Gro catalog number 10-040-CM) modified to contain 5% FBS, lOOug/mlPen/Strep (Cell-Gro catalog number 30-002-CL), and 0.3mg/ml L-Glutamine (Cell-Gro catalog number 25 -003 -CL). A 20,000 cells/ml solution was made for plating. Cells were plated in black Packard 96 well plates by placing 100 μl per well (2,000 cells per well). Table 3 below shows these and additional cell line data. The definitions of the cell types used are as follows: A549 is lung carcinoma; H1299 is non-small cell lung carcinoma; Hl 155 is non-small cell lung carcinoma; AsPC-I is pancreatic adenocarcinoma; Caov-3 is ovarian adenocarcinoma; COLO 205 is colorectal adenocarcinoma; DLD-I is colorectal adenocarcinoma; HCTl 16 is colorectal carcinoma; DU 145 is prostate carcinoma; ES-2 is ovarian clear cell carcinoma; H460 is large cell lung carcinoma; HELA is cervical adenocarcinoma; MIA PaCa-2 is pancreatice carcinoma; OVCAR-3 is ovarian adenocarcinoma; OVCAR8 is ovarian carcinoma; PC3 is prostate adenocarcinoma; SK-OV-3 is ovarian adenocarcinoma; SU86.86 is pancreatic carcinoma; TOV-21G is avarian clear cell carcinoma; U2OS is bone osteosarcoma; is ASPC-I is pancreatic adenocarcinoma; BXPC-3 is pabcreatic adenocarcinoma; HL60 is promyeloblast promyelocyte leukemia; K562 is bone marrow chronic myelogenous leukemia; L1210 is mouse lymphocytic leukemia; MOLT3 is T lymphoblast acute lymphoblastic leukemia; MOLT4 is T lymphoblast acute lymphoblastic leukemia; SW620 is colorectal adenocarcinoma; THP-I is monocyte acute monocytic leukemia; U937 is histiocytic lymphoma; and UACC-257 is melanoma. Cell Treatment with Compounds

[0333] Compounds and additional media were added 24 hours after cell plating. A compound master plate was created with concentrations 500 times greater than the final concentration added to the cells. All compound testing was done in duplicate using 6.3 fold dilutions starting with 1OmM. All outside wells (and 4 internal wells) were DMSO controls. Taxol and at least one additional control were run on all plates. Three microliters of the compound master plate were added to deep well blocks containing 750 μl of RPMI media. One hundred microliters were transferred from the compound/media deep well blocks to the plated cells resulting in a 500 fold dilution of the compounds. Cells were grown at 37⁰C, 5% CO₂ for 48 hours.

Photographic Image Analysis of Proliferation, Apoptosis and Death (PAD Assay) Cells to be analyzed by photography were fixed and stained. One hundred microliters of media were removed and 100 μl of 9.3% formamide was added to each well. Plates were left on the benchtop for 45 minutes. A staining solution containing 1.55 μl of lmg/ml DAPI added to 18.75ml PBS was warmed for 15 minutes at 37⁰C. The cells were aspirated prior to washing with 100 μl of PBS. Seventy microliters of PBS were aspirated and 170 μl of the DAPI solution were added to each well of fixed cells. Plates were left at room temperature for one hour then aspirated and washed twice with 100 μl of PBS. The stained cells were left at 4⁰C for a minimum of 16 hours before photographic analysis with Array Scan II (Cellomics). Analysis of the photographic images to determine numbers of live cells (proliferation), apoptotic cells and dead cells, were according to the methods described in U.S. Utiltiy Patent Application Serial No. 10/652,440, which incorporated herein in its entirety. Non-photographic Proliferation Analysis

[0334] Some cell plates were treated with Promega Cell titer Aqueous 1 kit (promega - VWR catalog number G3580). In this case, 48 hours after the test compound were added, 100 μl of media were removed and 20 μl of cell titer reagent were added to all wells. Plates were incubated at 37⁰C for 45 minutes prior to absorbance reads on the Wallac plate reader at 490nm for 0.1 sec/well. Results were similar to those obtained via the photographic analysis (described above).

Biological Example 7 Ubiquitination of p27 by SCF

Ubiquitination of p27 by SCF was assayed in a ubiquitin ligase assay essentially as described in US Patent No. 6,919,184 for screening ubiquitination of a ligase substrate and for modulators thereof.

Briefly, high protein binding plates (Corning 3922; Fisher # 07-200-591) were coated with 100 ng GST-p27 in TBS (Sigma), then aspirated. Reaction Buffer

5OmM Tris pH 7.5 (Trizma Base-Sigma T-8524) 7.5mM MgCl₂ (Magnesium Chloride - Sigma M-2393) 1.OmM DTT (DTT- Sigma D-9779) 2mM ATP (Roche Boehringer Mann Corp.- 635-316) 0.01% Tween20 0.02% BSA

50 μL of reaction buffer with 20ng GST-CyclinE/Cdk2 was added to each well, and this was incubated for 30 minutes. lOul of compound in diluted in DMSO was added to each well, followed by a 20ul volume of reaction buffer with 20ng El, 180ng Ubc3 and 40ng Skpl/Skp2, and a 20ul volume of reaction buffer with 60ng Rocl/Cull, 20ng his-Cksl and 200ng FLAG-Ubiquitin. This was incubated for 90 minutes. The plates were then washed with IX TBST and lOOul TBST-0.25% BSA with anti-FLAG Ab (1 :30,000) and anti-mouse IgG-HRP Ab (1 : 150,000 GE Healthcare) were added and incubate for 60 minutes. The plates were washed again with IX TBST and Lumino substrate (Pierce) was added. Luminescence was read on a BMG Fluostar plate reader. The following compounds of the invention were assayed as above. In the following table, IC₅₀ values are listed by range, where A = < 5 μM, B= 5-20 μM, and C = > 20 μM.

Biological Example 8 Ubiquibitination of PCNA by Radl8

[0335] Ubiquitination of PCNA by Radl8 was assayed in a similar fashion as described above for ubiquitination of p27.

[0336] Briefly, high protein binding plates were coated with 200ng/well of His-PCNA in TBS (sigma T6664), then aspirated. 80 μL of reaction buffer and 200 ng FLAG-ubiquitin were added to each well, followed by 10 μL of compound in 100% DMSO (varying concentrations, 100 % DMSO used as a control). A 10 μL mixture of El (hEl- 10 ng/well), E2 (Rad6-25 ng/well) and E3 (Radl8-50 ng/well) were then added to the mixture and this was incubated for 1 hour. The plates were then washed 5 times with TBST and Incubated at room temp for 1 hour with lOOμl/well of anti-Flag-HRP at 1 :10,000 (Sigma A8592) in 0.25% BSA IXPBS, washed again and treated with Immobilon Western Chemiluminescence HRP Substrate (Millipore WKBLS0500). 100 % DMSO in place of compound solution and plates starting with TBS with no PCNA substrate were used as controls to show activity and specificity of compounds. Fluorescence was read on a BMG Fluostar plate reader.

[0337] The following compounds of the invention were assayed as above. In the following table, IC₅₀ values are listed by range, where A = < 5 μM, B= 5-20 μM, and C = > 20 μM.

Biological Example 9 Ubiquibitination of Oligonucleasome by BMI-1/Ringlb

Ubiquitination of Oligonucleasome by BMI-1/Ringlb was assayed in a similar fashion as described above.

Briefly, high protein binding plates were coated with oligonucleasome (isolated from HELA cells), then washed with TBST. 80 μl of reaction buffer with 200 ng FLAG-ubiquitin were added to each well, followed by 10 μl compound solution. A 10 μl aliquot containing 10 ng El, 25 ng E2(UbcH5c), and 50 ng Ring lb/Bmi-1 were then added to each well. After incubation, the plates were washed with TBST and lOOμl/well of antibody mix (1 :30,000 anti- Flag (Sigma) and 1/150,000 of anti-Mouse-HRP (GE Healthcare) in 0.25% BSA) was added to each well. Following antibody incubation, the plates were washed with TBST, Lumino substrate was added, and the plates were read.

The following compounds of the invention were assayed as above. In the following table, IC50 values are listed by range, where A = < 5 μM, B= 5-20 μM, and C = > 20 μM.

Claims

We Claim:

1. A compound according to the formula,

or pharmaceutically acceptable salts, hydrates, solvates, polymorphs, atrophisomers, N-oxides, and prodrugs thereof, wherein n is 0, 1, 2, 3, or 4; L' is a covalent bond or -SO₂-; A is -C(X)(Y)-, -N(Y)-, or -O-, wherein X is H, -C(O)R₈, or -OH; and

Y is -H, -NO₂, -OH, -CN, halo, -Z, Ci-C₆ alkyl, Ci-C₆ alkoxy, -NH₂, mono- or (Ii-(C₁-C₆ alkyl) amino, -C(O)-OR₉, -C(O)R₉, -N(R₈)-Z, Ci-C₆ alkyl-aryl, Ci-C₆ alkyl-heterocyclyl, C₁-C₆ alkyl-heteroaryl, C₀-C₆ alkyl-C(O)-aryl, C₀-C₆ alkyl-C(O)-heterocyclyl, C₀-C₆ alkyl-C(O)-heteroaryl, -O-aryl, C₃-C₈-cycloalkyl, heterocyclyl, or -Zi-S(O)₂-Z₂ where

Zi and Z₂ are independently aryl or heteroaryl, wherein each of the aryl, heterocyclyl and heteroaryl is optionally substituted with 1 to 3 groups selected from -H, Ci-C₆ alkyl, Ci-C₆ alkoxy, mono- to per-halogenated Ci-C₆ alkyl, -C(O)-OR₉, -N(R₉)-C(O)R₉, -O-(halo Ci-C₆ alkyl), -NH₂, mono- or di-(Ci-C₆ alkyl) amino, -NO₂, halo, or -CN; or X and Y together with the atom to which they are attached, are oxo, a C₃-C₈ cycloalkyl or heterocyclyl wherein the cycloalkyl or heterocyclyl is optionally substituted by one to four R⁴ groups;

Ri and R₂ are independently -H, -NO₂, -OH, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, mono- to per-halogenated Ci-C₆ alkyl, heterocyclyl, heterocyclyl-O-aryl, -NH₂, mono- or di-(Ci-C₆ alkyl) amino, -C(O)-OR₉, -N(R₈)-Z, -0-Z, -S-Z, -SO₂-Z, -SO₂-NH-Z, -N(R₈)-Z, aryl, or heteroaryl, wherein each of the aryl and heteroaryl is optionally substituted with 1 to 3 groups selected from -H, Ci-C₆ alkyl, Ci-C₆ alkoxy, mono- to per-halogenated Ci-C₆ alkyl, -NH₂, mono- or di-(Ci-C₆ alkyl) amino, -NO₂, halo, or -CN; or Ri and R₂ together with the carbon atoms to which they are attached form an aryl group optionally substituted with 1 to 3 groups selected from -OH, halo, Ci-C₆-alkyl, Ci-C₆-alkoxy, -NH₂, and -NO₂; each R₃ is independently halo or Ci-C₆ alkyl, or two R3 attached to adjacent carbon atoms, together with the atoms to which they are attached, form a fused aryl or heteroaryl wherein the aryl or heteroaryl is optionally substituted by one to four R⁴ groups; or Y and R3 attached to adjacent carbon atoms, together with the atoms to which they are attached, form a fused aryl or heteroaryl and X is absent, wherein the aryl or heteroaryl is optionally substituted by one to four R⁴ groups; each R₄ is independently halo, Ci-C₆ alkyl, oxo, or Ci-C₆ alkoxy; R₈ is -H, Ci-C₆ alkyl, or Ci-C₆ alkoxy; R9 is -H, Ci-C₆ alkyl or C₃-C₆ cycloalkyl; and each Z is independently -Ci-C₆ alkyl-O-C(O)-Ci-C₆ alkyl, aryl, or heteroaryl wherein each of the aryl and heteroaryl is optionally substituted with one to four R₅ groups, wherein each R₅ is independently -NO₂, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, -NH₂, mono- or di-(Ci-C₆ alkyl) amino, oxo, or -C(O)ORg, provided the compound is not a compound listed in Tables 2 and 2A.

2. The compound of claim 1, of the formula,

3. The compound of claim 2, of the formula,

4. The compound of claim 3, wherein Z is aryl or heteroaryl wherein the aryl and heteroaryl is optionally substituted with one to four R5 groups, wherein each R5 is independently -NO₂, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, -NH₂, mono- or (Ii-(C₁-C₆ alkyl) amino, or -C(O)OR₉.

5. The compound of claim 4, wherein Z is aryl optionally substituted with one or two R₅ groups.

6. The compound of claim 4, wherein Z is phenyl optionally substituted with one or two R₅ groups.

7. The compound of claim 4, wherein Z is quinolinyl optionally substituted with one or two R₅ groups.

8. The compound of claim 7, wherein Z is pyridyl optionally substituted with one or two R5 groups.

9. The compound of claim 3, of the formula,

10. The compound of claim 9, wherein

Y is aryl or heteroaryl wherein the aryl and heteroaryl is optionally substituted with one to four R₅ groups, wherein each R₅ is independently -NO₂, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, -NH₂, mono- or (Ii-(C₁-C₆ alkyl) amino, or -C(O)OR₉.

11. The compound of claim 9, wherein Z is aryl optionally substituted with one or two R5 groups.

12. The compound of claim 9, wherein Z is heteroaryl optionally substituted with one or two R₅ groups.

13. The compound of claim 9, of one of the formulas,

XIX XX. wherein p is 0, 1, 2, 3, or 4; and each R₆ is independently -NO₂, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, -NH₂, mono- or di-(Ci-C₆ alkyl) amino, or -C(O)ORg.

14. The compound of claim 13, wherein p is O or 1.

15. The compound of claim 13, wherein each R₆ is independently -NO₂, -CN, halo, Ci-C₆ alkyl, or Ci-C₆ alkoxy, and p is 1 or 2.

16. The compound of claim 13, wherein Z is phenyl optionally substituted with one or two R₅ groups.

17. The compound of claim 13, wherein Z is pyridyl or quinolinyl optionally substituted with one or two R₅ groups.

18. The compound of claim 2, of the formula,

19. The compound of claim 18, wherein Y is aryl or heteroaryl wherein the aryl and heteroaryl is optionally substituted with one to four R₅ groups, wherein each R₅ is independently -NO₂, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, -NH₂, mono- or di-(Ci-C₆ alkyl) amino, or -C(O)OR₉.

20. The compound of claim 18, wherein Z is aryl optionally substituted with one or two R₅ groups.

21. The compound of claim 18, wherein Z is heteroaryl optionally substituted with one or two R5 groups.

22. The compound of claim 2, of the formula,

23. The compound of claim 22, wherein

24. The compound of claim 22, wherein Z is aryl optionally substituted with one or two R₅ groups.

25. The compound of claim 22, wherein Z is heteroaryl optionally substituted with one or two R5 groups.

26. The compound of claim 22, of the formula,

XXII wherein q is O, 1, 2, 3, or 4; and each R₇ is independently -NO₂, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, -NH₂, mono- or (Ii-(C₁-C₆ alkyl) amino, or -C(O)OR₉.

27. The compound of claim 26, wherein each R₇ is independently -NO₂, -CN, halo, Ci-C₆ alkyl, or Ci-C₆ alkoxy, and q is 1 or 2.

28. The compound of claim 26, wherein Z is phenyl optionally substituted with one or two R5 groups, wherein each R₅ is independently -NO₂, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, -NH₂, mono- or di-(Ci-C₆ alkyl) amino, or -C(O)OR₉.

29. The compound of claim 26, wherein Z is quinolinyl optionally substituted with one or two R5 groups, wherein each R₅ is independently -NO₂, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, -NH₂, mono- or di-(Ci-C₆ alkyl) amino, or -C(O)OR₉.

30. The compound of claim 2, of the formula,

wherein m is O, 1, 2, 3, or 4.

31. The compound of claim 30, wherein m is 0, 1 , or 2.

32. The compound of claim 30, wherein Z is aryl or heteroaryl wherein the aryl and heteroaryl is optionally substituted with one to four R₅ groups, wherein each R₅ is independently -NO₂, -CN, halo, Ci-C₆ alkyl, Ci-C₆ alkoxy, -NH₂, mono- or di-(Ci-C₆ alkyl) amino, or -C(O)OR₉.

33. The compound of claim 32, wherein Z is phenyl optionally substituted with one or two R₅ groups.

34. The compound of claim 32, wherein Z is pyridyl or quinolinyl optionally substituted with one or two R₅ groups.

35. The compound of claim 1 which is,

Name

2-(4-(benzo[c][l,2,5]oxadiazol-4-ylsulfonyl)piperazin-l-yl)nicotinonitrile;

6-(4-(benzo[c][l,2,5]oxadiazol-4-ylsulfonyl)piperazin-l-yl)nicotinonitrile; 4-(4-(pyridin-4-yl)piperazin-l-ylsulfonyl)benzo[c][l,2,5]oxadiazole; 4-(4-(3,4-dimethylphenyl)piperazin-l-ylsulfonyl)benzo[c][l,2,5]oxadiazole; l-(benzo[c][l,2,5]oxadiazol-4-ylsulfonyl)-4-(4-chlorophenyl)piperidin-4-ol; l-(7-chlorobenzo[c][l,2,5]oxadiazol-4-ylsulfonyl)-4-(4-chlorophenyl)piperidin-4-ol;

4-(4-chlorophenyl)-l-(7-(quinolin-8-ylthio)benzo[c][l,2,5]oxadiazol-4-ylsulfonyl)piperidin-4- ol; l-(l-(7-chlorobenzo[c][l,2,5]oxadiazol-4-ylsulfonyl)piperidin-4-yl)-lH-benzo[d]imidazol-2(3 H)-one; l-(l-(7-(quinolin-8-ylthio)benzo[c][l,2,5]oxadiazol-4-ylsulfonyl)piperidin-4-yl)-lH-benzo[d]i midazol-2(3H)-one;

4-(3,5-dimethylpiperidin-l-ylsulfonyl)-7-(quinolin-8-ylthio)benzo[c][l,2,5]oxadiazole; 4-(2,6-dimethylmorpholinosulfonyl)-7-(quinolin-8-ylthio)benzo[c][l,2,5]oxadiazole; 4-(2,6-dimethylmorpholinosulfonyl)-5-(quinolin-8-ylthio)benzo[c][l,2,5]oxadiazole; 1 -(I -(7-chlorobenzo[c] [ 1 ,2,5]oxadiazol-4-ylsulfonyl)-4-phenylpiperidin-4-yl)ethanone;

4-(4-chlorophenyl)-l-(7-(quinolin-2-ylthio)benzo[c][l,2,5]oxadiazol-4-ylsulfonyl)piperidin-4- ol;

4-(2-methoxyphenylthio)-7-(4-phenylpiperazin- 1 -ylsulfonyl)benzo[c] [ 1 ,2,5]oxadiazole; 4-(4-phenylpiperazin- 1 -ylsulfonyl)-7-(quinolin-8-yloxy)benzo[c] [ 1 ,2,5]oxadiazole; 4-(3H-spiro[isobenzoflιran- 1 ,4'-piperidine]- 1 '-ylsulfonyl)-7-chlorobenzo[c] [1 ,2,5]oxadiazole;

4-(4-chlorophenyl)-l-(7-(pyridin-4-ylthio)benzo[c][l,2,5]oxadiazol-4-ylsulfonyl)piperidin-4-o i r-(7-chlorobenzo[c][l,2,5]oxadiazol-4-ylsulfonyl)-3H-spiro[isobenzofuran-l,4'-piperidin]-3- one;

4-(4-(4-methoxyphenyl)piperidin- 1 -ylsulfonyl)-7-(2-methoxyphenylthio)benzo[c] [ 1 ,2,5]oxadi azole;

4-(4-methoxyphenyl)-l-(7-(2-methoxyphenylthio)benzo[c][l,2,5]oxadiazol-4-ylsulfonyl)piper idin-4-ol;

4-(4-methoxyphenyl)-l-(7-(quinolin-8-ylthio)benzo[c][l,2,5]oxadiazol-4-ylsulfonyl)piperidin -4-ol;

4-(4-(4-chlorophenyl)piperazin-l-ylsulfonyl)-7-(2-methoxyphenylthio)benzo[c][l,2,5]oxadiaz ole; 4-(4-(4-chlorophenyl)piperidin-l-ylsulfonyl)-7-(l,2,3,4-tetrahydroquinolin-8-ylthio)benzo[c][ l,2,5]oxadiazole;

4-(4-chlorophenyl)-l-(7-(l,2,3,4-tetrahydroquinolin-8-ylthio)benzo[c][l,2,5]oxadiazol-4-ylsul fonyl)piperidin-4-ol;

4-(4-(4-chlorophenyl)piperidin-l-ylsulfonyl)-7-(4-methoxyphenylthio)benzo[c][l,2,5]oxadiaz ole;

4-(4-(4-chlorophenyl)piperidin-l-ylsulfonyl)-7-(3-methoxyphenylthio)benzo[c][l,2,5]oxadiaz ole;

4-(4-methoxyphenyl)-l-(7-(4-methoxyphenylthio)benzo[c][l,2,5]oxadiazol-4-ylsulfonyl)piper idin-4-ol;

4-(4-methoxyphenyl)-l-(7-(3-methoxyphenylthio)benzo[c][l,2,5]oxadiazol-4-ylsulfonyl)piper idin-4-ol;

4-(7-chloro-3,4-dihydroisoquinolin-2(lH)-ylsulfonyl)-7-(3-methoxyphenylthio)benzo[c][l,2,5 ]oxadiazole;

4-(6-chloro-3,4-dihydroisoquinolin-2(lH)-ylsulfonyl)-7-(2-methoxyphenylthio)benzo[c][l,2,5 Joxadiazole;

4-(2,6-dimethylmorpholinosulfonyl)-7-(4-methoxyphenylthio)benzo[c] [ 1 ,2,5]oxadiazole; 4-(2,6-dimethylmorpholinosulfonyl)-7-(3-methoxyphenylthio)benzo[c] [ 1 ,2,5]oxadiazole;

4-(7-chloro-3,4-dihydroisoquinolin-2(lH)-ylsulfonyl)-7-(4-methoxyphenylthio)benzo[c][l,2,5 Joxadiazole;

4-(6-chloro-3,4-dihydroisoquinolin-2(lH)-ylsulfonyl)-7-(4-methoxyphenylthio)benzo[c][l,2,5 ]oxadiazole;

4-(6-chloro-3,4-dihydroisoquinolin-2(lH)-ylsulfonyl)-7-(3-methoxyphenylthio)benzo[c][l,2,5 Joxadiazole;

4-(7-chloro-3,4-dihydroisoquinolin-2(lH)-ylsulfonyl)-7-(quinolin-8-ylthio)benzo[c][l,2,5]oxa diazole;

4-(6-chloro-3,4-dihydroisoquinolin-2(lH)-ylsulfonyl)-7-(quinolin-8-ylthio)benzo[c][l,2,5]oxa diazole;

4-(3,5-dimethylpiperidin-l-ylsulfonyl)-7-(4-methoxyphenylthio)benzo[c][l,2,5]oxadiazole; 4-(3,5-dimethylpiperidin-l-ylsulfonyl)-7-(3-methoxyphenylthio)benzo[c][l,2,5]oxadiazole; 4-(3,5-dimethylpiperidin- 1 -ylsulfonyl)-7-(3-ethoxyphenylthio)benzo[c] [ 1 ,2,5]oxadiazole;

4-(4-(4-chlorophenyl)piperazin-l-ylsulfonyl)-7-(4-methoxyphenylthio)benzo[c][l,2,5]oxadiaz ole; 4-(7-chloro-3,4-dihydroisoquinolin-2(lH)-ylsulfonyl)-7-(pyridin-2-ylthio)benzo[c][l,2,5]oxad iazole;

4-(6-chloro-3,4-dihydroisoquinolin-2(lH)-ylsulfonyl)-7-(pyridin-2-ylthio)benzo[c][l,2,5]oxad iazole;

4-(3,5-dimethylpiperidin- 1 -ylsulfonyl)-7-(4-ethoxyphenylthio)benzo[c] [ 1 ,2,5]oxadiazole;

4-(7-chloro-3,4-dihydroisoquinolin-2(lH)-ylsulfonyl)-7-(3-ethoxyphenylthio)benzo[c][l,2,5]o xadiazole;

4-(7-chloro-3,4-dihydroisoquinolin-2(lH)-ylsulfonyl)-7-(4-ethoxyphenylthio)benzo[c][l,2,5]o xadiazole;

4-(3,4-dihydro- 1 ,8-naphthyridin- 1 (2H)-ylsulfonyl)-7-(2-methoxyphenylthio)benzo[c] [ 1 ,2,5]o xadiazole;

4-(4-(4-chlorophenyl)piperidin- 1 -ylsulfonyl)-7-(3-ethoxyphenylthio)benzo[c] [ 1 ,2,5]oxadiazol e;

4-(2,6-dimethylmorpholinosulfonyl)-7-(3-ethoxyphenylthio)benzo[c][l,2,5]oxadiazole;

4-(4-(4-methoxyphenyl)piperidin- 1 -ylsulfonyl)-7-(4-methoxyphenylthio)benzo[c] [ 1 ,2,5]oxadi azole;

4-(4-(4-methoxyphenyl)piperidin- 1 -ylsulfonyl)-7-(3-methoxyphenylthio)benzo[c] [ 1 ,2,5]oxadi azole;

4-(4-(4-chlorophenyl)piperazin-l-ylsulfonyl)-7-(3-methoxyphenylthio)benzo[c][l,2,5]oxadiaz ole;

4-(4-(4-chlorophenyl)piperazin-l-ylsulfonyl)-7-(3-ethoxyphenylthio)benzo[c][l,2,5]oxadiazol e;

4-(4-(4-chlorophenyl)piperidin- 1 -ylsulfonyl)-7-(4-ethoxyphenylthio)benzo[c] [ 1 ,2,5]oxadiazol e; l-(7-(3-ethoxyphenylthio)benzo[c][l,2,5]oxadiazol-4-ylsulfonyl)-4-(4-methoxyphenyl)piperid in-4-ol; l-(7-(4-ethoxyphenylthio)benzo[c][l,2,5]oxadiazol-4-ylsulfonyl)-4-(4-methoxyphenyl)piperid in-4-ol;

4-(3,5-dimethylpiperidin-l-ylsulfonyl)-7-(pyridin-2-ylthio)benzo[c][l,2,5]oxadiazole;

4-(2,6-dimethylmorpholinosulfonyl)-7-(pyridin-2-ylthio)benzo[c][l,2,5]oxadiazole;

4-(2,6-dimethylmorpholinosulfonyl)-7-(4-ethoxyphenylthio)benzo[c][l,2,5]oxadiazole;

4-(7-chloro-3,4-dihydroisoquinolin-2(lH)-ylsulfonyl)-7-(p-tolylthio)benzo[c][l,2,5]oxadiazol e; l-(7-(4-methoxyphenylthio)benzo[c][l,2,5]oxadiazol-4-ylsulfonyl)piperidin-4-ol; 4-(3,5-dimethylpiperidin-l-ylsulfonyl)-7-(p-tolylthio)benzo[c][l,2,5]oxadiazole; l-(7-(p-tolylthio)benzo[c][l,2,5]oxadiazol-4-ylsulfonyl)piperidin-4-ol; 4-(3,5-dimethylpiperidin-l-ylsulfonyl)-7-(4-ethylphenylthio)benzo[c][l,2,5]oxadiazole;

4-(7-chloro-3,4-dihydroisoquinolin-2(lH)-ylsulfonyl)-7-(4-ethylphenylthio)benzo[c][l,2,5]ox adiazole;

4-(7-chloro-3,4-dihydroisoquinolin-2(lH)-ylsulfonyl)-7-(naphthalen-l-ylthio)benzo[c][l,2,5] oxadiazole;

4-(4-(4-chlorophenyl)piperazin-l-ylsulfonyl)-7-(4-ethoxyphenylthio)benzo[c][l,2,5]oxadiazol e;

4-(piperidin- 1 -ylsulfonyl)-7-(pyridin-2-ylthio)benzo[c] [ 1 ,2,5]oxadiazole; 4-(2-methoxyphenylthio)-7-(piperidin- 1 -ylsulfonyl)benzo[c] [ 1 ,2,5]oxadiazole; 4-(4-methoxyphenylthio)-7-(piperidin- 1 -ylsulfonyl)benzo[c] [ 1 ,2,5]oxadiazole; 4-(3-methoxyphenylthio)-7-(piperidin- 1 -ylsulfonyl)benzo[c] [ 1 ,2,5]oxadiazole; l-(7-(2-methoxyphenylthio)benzo[c][l,2,5]oxadiazol-4-ylsulfonyl)piperidin-4-ol; 4-(3,5-dimethylpiperidin- 1 -ylsulfonyl)-7-(naphthalen- 1 -ylthio)benzo[c] [1 ,2,5]oxadiazole; 4-(2,6-dimethylmorpholinosulfonyl)-7-(4-ethylphenylthio)benzo[c][l,2,5]oxadiazole; 4-chloro-7-(7-chloro-3,4-dihydroisoquinolin-2(lH)-ylsulfonyl)benzo[c][l,2,5]oxadiazole;

4-(7-chloro-3,4-dihydroisoquinolin-2(lH)-ylsulfonyl)-7-(2-methoxyphenylthio)benzo[c][l,2,5 ]oxadiazole;

4-chloro-7-(6-chloro-3,4-dihydroisoquinolin-2(lH)-ylsulfonyl)benzo[c][l,2,5]oxadiazole;

4-(4-(4-chlorophenyl)piperidin-l-ylsulfonyl)-7-(2-methoxyphenylthio)benzo[c][l,2,5]oxadiaz ole;

4-(3,5-dimethylpiperidin-l-ylsulfonyl)-7-(2-methoxyphenylthio)benzo[c][l,2,5]oxadiazole;

4-(4-chlorophenyl)-l-(7-(quinolin-8-yloxy)benzo[c][l,2,5]oxadiazol-4-ylsulfonyl)piperidin-4- oU

4-(4-chlorophenyl)-l-(7-(quinolin-8-yl)benzo[c][l,2,5]oxadiazol-4-ylsulfonyl)piperidin-4-ol;

4-(4-chlorophenyl)-l-(7-(quinolin-8-ylsulfonyl)benzo[c][l,2,5]oxadiazol-4-ylsulfonyl)piperidi n-4-ol; 4-(4-chlorophenyl)-l-(7-(quinolin-8-ylamino)benzo[c][l,2,5]oxadiazol-4-ylsulfonyl)piperidin -4-ol;

4-(4-(4-chlorophenyl)piperidin- 1 -ylsulfonyl)-7-(quinolin-8-ylthio)benzo[c] [ 1 ,2,5]oxadiazole;

4-phenyl-l-(7-(quinolin-8-ylthio)benzo[c][l,2,5]oxadiazol-4-ylsulfonyl)piperidin-4-ol;

4-(2-chlorophenyl)-l-(7-(quinolin-8-ylthio)benzo[c][l,2,5]oxadiazol-4-ylsulfonyl)piperidin-4- ol;

4-(3-chlorophenyl)-l-(7-(quinolin-8-ylthio)benzo[c][l,2,5]oxadiazol-4-ylsulfonyl)piperidin-4- ol;

4-(4-fluorophenyl)-l-(7-(quinolin-8-ylthio)benzo[c][l,2,5]oxadiazol-4-ylsulfonyl)piperidin-4- ol;

4-(3-fluorophenyl)-l-(7-(quinolin-8-ylthio)benzo[c][l,2,5]oxadiazol-4-ylsulfonyl)piperidin-4- ol;

4-(4-(4-chlorophenyl)piperidin- 1 -ylsulfonyl)-7-(pyridin-2-ylthio)benzo[c] [ 1 ,2,5]oxadiazole;

4-(4-chlorophenyl)-l-(7-(pyridin-2-ylthio)benzo[c][l,2,5]oxadiazol-4-ylsulfonyl)piperidin-4-o 1;

7-(4-(4-chlorophenyl)piperidin-l-yl)-N-(quinolin-8-yl)benzo[c][l,2,5]oxadiazole-4-sulfonami de;

4-(2,6-dimethylmorpholinosulfonyl)-7-(2-methoxyphenylthio)benzo[c] [ 1 ,2,5]oxadiazole;

7-(4-(4-chlorophenyl)-4-hydroxypiperidin-l-yl)-N-(quinolin-8-yl)benzo[c][l,2,5]oxadiazole-4 -sulfonamide;

4-(4-chlorophenyl)-l-(7-(2-methoxyphenylthio)benzo[c][l,2,5]oxadiazol-4-ylsulfonyl)piperid in-4-ol;

4-(4-(4-chlorophenyl)piperazin-l-ylsulfonyl)-7-(quinolin-8-ylthio)benzo[c][l,2,5]oxadiazole;

1 -(7-(quinolin-8-ylthio)benzo[c] [ 1 ,2,5]oxadiazol-4-ylsulfonyl)piperidin-4-ol;

4-(piperidin-l-ylsulfonyl)-7-(quinolin-8-ylthio)benzo[c][l,2,5]oxadiazole; or

4-(2-fluorophenyl)-l-(7-(quinolin-8-ylthio)benzo[c][l,2,5]oxadiazol-4-ylsulfonyl)piperidin-4- ol.

36. A pharmaceutical composition comprising, together with a pharmaceutically acceptable carrier, diluent, or excipient, a compound of claim 1.

37. A method of inhibiting ubiquitination in a cell comprising contacting the cell in which inhibition of ubiquitination is desired with a compound of claim 1.

38. A method of inhibiting ubiquitination in a cell comprising contacting the cell in which inhibition of ubiquitination is desired with a pharmaceutical composition of claim 36.

39. The method of claim 37 or 38, wherein ubiquitin ligase activity of p27/SCF, TRAF6, Radl8, or BMI-I is inhibited.

40. A method of treating cell proliferative diseases or conditions comprising administering to a patient an effective amount of a compound of claim 1.

41. A method of treating cell proliferative diseases or conditions comprising administering to a patient an effective amount of a pharmaceutical composition of claim 36.

42. The method of claim 40 or 41, wherein the cell proliferative disease or condition is psoriasis, keloid scarring, or cancers of the breast, immune system, bone, nervous system, brain, blood, lymphatic system, or skin.

43. A method of treating conditions or diseases that involve TRAF6 comprising administering to a patient an effective amount of a compound of claim 1.

44. A method of treating conditions or diseases that involve TRAF6 comprising administering to a patient an effective amount of a pharmaceutical composition of claim 36.

45. The method of claim 43 or 44, wherein the conditions or diseases that involve TRAF6 is cancer, inflammation, adaptive immunity, innate immunity, bone metabolism, LPS-induced angiogenesis, osteoporosis, osteopinneal diseases, lymph node development, mammary gland development, skin development, or central nervous system development.

46. A method of treating conditions or diseases that involve p27/SCF comprising administering to a patient an effective amount of a compound of claim 1.

47. A method of treating conditions or diseases that involve p27/SCF comprising administering to a patient an effective amount of a pharmaceutical composition of claim 36.

48. The method of claim 46 or 47, wherein the conditions or diseases that involve p27/SCF is cancer, inflammation, adaptive immunity, innate immunity, bone metabolism, LPS-induced angiogenesis, osteoporosis, osteopinneal diseases, lymph node development, mammary gland development, skin development, or central nervous system development.

49. A method of treating conditions or diseases that involve Radl8 comprising administering to a patient an effective amount of a compound of claim 1.

50. A method of treating conditions or diseases that involve Radl8 comprising administering to a patient an effective amount of a pharmaceutical composition of claim 36.

51. The method of claim 49 or 50, wherein the conditions or diseases that involve Radl 8 is cancer, inflammation, adaptive immunity, innate immunity, bone metabolism, LPS-induced angiogenesis, osteoporosis, osteopinneal diseases, lymph node development, mammary gland development, skin development, or central nervous system development.

52. A method of treating conditions or diseases that involve BMI-I comprising administering to a patient an effective amount of a compound of claim 1.

53. A method of treating conditions or diseases that involve BMI-I comprising administering to a patient an effective amount of a pharmaceutical composition of claim 36.

54. The method of claim 52 or 53, wherein the conditions or diseases that involve BMI-I is cancer, inflammation, adaptive immunity, innate immunity, bone metabolism, LPS-induced angiogenesis, osteoporosis, osteopinneal diseases, lymph node development, mammary gland development, skin development, or central nervous system development.

55. A compound of the formula,

(XL) wherein,

R is Ci-C₆ alkyl, Ci-C₆ haloalkyl, aryl, or heteroaryl, wherein R is optionally substituted with one to four R³ groups;

Li is L₃, -Ci-C₆ alkyl-, -Ci-C₆ alkyl-L₃-, -L₃-Ci-C₆ alkyl-, or -L₃-Ci-C₆ alkyl-L₃, wherein each L₃ is independently -C(O)N(R₄)-, -N(R₄)C(O)-, -OC(O)-, -C(O)O-, -N(R₄)C(O)N(R₄)-, -OC(O)N(R₄)-, -N(R₄)C(O)O-, -OC(O)O-, -C(O)-, -S(O)-, -S(O)₂-, -S(O)₂N(R₄)-, or -N(R₄)S(O)₂-;

Ri is aryl or heteroaryl, wherein Ri is optionally substituted with one to four R groups; L₂ is -C(O)N(R₄)-, -N(R₄)C(O)-, -OC(O)-, -C(O)O-, -N(R₄)C(O)N(R₄)-, -OC(O)N(R₄)-, -N(R₄)C(O)O-, -OC(O)O-, -C(O)-, -S(O)-, -S(O)₂-, -S(O)₂N(R₄)-, or -N(R₄)S(O)₂-;

-N(R₆)S(O)₂R₆₅-OS(O)₂R₆, -C(O)R₆, -C(O)OR₆, -C(O)N(Re)₂, -OC(O)R₆, -OC(O)OR₆, -OC(O)N(R₆)₂, -N(R₆)C(O)R₆, -N(R₆)C(O)OR⁶, -N(R₆)C(O)N(Re)₂, -CN, -NO₂, -Ci-C₆ alkyl, -C₁-C₆ haloalkyl, -Ci-C₆ alkyl-OR₆, or -Ci-C₆ alkyl-N(R₆)₂, wherein each R₆ is independently-H, Ci-C₆ alkyl, Ci-C₆ haloalkyl, C₃-Cg cycloalkyl, heterocyclyl, aryl, or heteroaryl; or any two R₃ on adjacent carbon atoms, taken together with the atoms to which they are attached, form a fused C₃-Cs cycloalkyl, heterocyclyl, aryl, or heteroaryl group; and each R₄ is independently -H, Ci-C₆ alkyl, or Ci-C₆ haloalkyl, provided the compound is not

56. The compound of claim 55, of the formula,

57. The compound of claim 56, wherein R is Ci-C₆ alkyl.

58. The compound of claim 56, wherein R is phenyl optionally substituted with one to four R3 groups.

59. The compound of claim 56, wherein Li is -Ci-C₆ alkyl-L₃- or -L₃-Ci-C₆ alkyl-, wherein each L₃ is independently -C(O)N(R₄)-, -N(R₄)C(O)-, -S(O)₂N(R₄)-, or -N(R₄)S(O)₂-.

60. The compound of claim 56, wherein L₂ is -N(R₄)C(O)N(R₄)-, -OC(O)N(R₄)-, -N(R₄)C(O)O-, -OC(O)O-.

61. A compound of the formula,

(LX) wherein m and n are independently O, 1, 2, 3, 4, or 5;

R₃, R₄, and R₅ are each independently halo, -OR₆, -N(Re)₂, -S(R₆), -S(O)₂R₆, -S(O)₂N(Rδ)₂, -S(O)₂OR₆, -N(R₆)S(O)₂R₆, -OS(O)₂R₆, -C(O)R₆, -C(O)OR₆, -C(O)N(R₆),, -OC(O)R₆, -OC(O)OR₆, -OC(O)N(Rg)₂, -N(R₆)C(O)R₆, -N(R₆)C(O)OR⁶, -N(R₆)C(O)N(Rg)₂, -CN, -NO₂, -Ci-C₆ alkyl, -Ci-C₆ haloalkyl, -Ci-C₆ alkyl-ORδ, or -Ci-C₆ alkyl-N(R₆)₂, wherein each R₆ is independently-H, Ci-C₆ alkyl, Ci-C₆ haloalkyl, C₃-Cg cycloalkyl, heterocyclyl, aryl, or heteroaryl, provided the compound is not Structure Name

(2-((4-hydroxybenzyl)(4-methoxybenzyl)amino)-4- phenylthiazol-5-yl)(phenyl)methanone;

(2-((4-hydroxybenzyl)(4-methoxyphenethyl)amino)-

4-(4-methoxyphenyl)thiazol-5-yl)(4- methoxyphenyl)methanone;

(2-((4-hydroxybenzyl)(4-methoxybenzyl)amino)-4-

(4-methoxyphenyl)thiazol-5-yl)(4- methoxyphenyl)methanone;

(2-(butyl(4-hydroxybenzyl)amino)-4-phenylthiazol-5- yl)(phenyl)methanone; or

(2-amino-4-phenylthiazol-5-yl)(phenyl)methanone.

62. The compound of claim 61, wherein Ri and R₂ are independently -Ci-C₆ alkyl-aryl or -Ci-C₆ alkyl-heteroaryl, wherein each is optionally substituted with one to four groups which are independenly halo, -OR₈, -N(Rg)₂, -S(R₈), -S(O)₂R₈, -S(O)²N(R₈)₂, -C(O)R₈, -C(O)OR₈, -C(O)N(Rs)₂, -CN, -NO₂, -Ci-C₆ alkyl, or - Ci-C₆ haloalkyl, wherein each R₈ is independently -H, Ci-C₆ alkyl.

63. A composition comprising, together with a pharmaceutically acceptable carrier, diluent, or excipient, a compound of claim 55 or 61 or a compound selected from

(2-((4-hydroxybenzyl)(4-methoxybenzyl)amino)-4-phenylthiazol-5- yl)(phenyl)methanone; (2-((4-hydroxybenzyl)(4-methoxyphenethyl)amino)-4-(4-methoxyphenyl)thiazol-5-yl)(4- methoxyphenyl)methanone;

(2-(butyl(4-hydroxybenzyl)amino)-4-phenylthiazol-5-yl)(phenyl)methanone; or

(2-amino-4-phenylthiazol-5-yl)(phenyl)methanone,

64. A method of inhibiting ubiquitination in a cell comprising contacting the cell in which inhibition of ubiquitination is desired with a compound of claim 55 or 61 or a compound selected from

(2-((4-hydroxybenzyl)(4-methoxybenzyl)amino)-4-phenylthiazol-5- yl)(phenyl)methanone;

(2-(butyl(4-hydroxybenzyl)amino)-4-phenylthiazol-5-yl)(phenyl)methanone; or

(2-amino-4-phenylthiazol-5-yl)(phenyl)methanone,

65. A method of inhibiting ubiquitination in a cell comprising contacting the cell in which inhibition of ubiquitination is desired with a pharmaceutical composition of claim 63.

66. The method of claim 64, wherein ubiquitin ligase activity of p27/SCF, TRAF6, Radl8, or BMI-I is inhibited.

67. The method of claim 65, wherein ubiquitin ligase activity of p27/SCF, TRAF6, Radl8, or BMI-I is inhibited.

68. A method of treating cell proliferative diseases or conditions comprising administering to a patient an effective amount of a compound of claim 55 or 61 or a compound selected from

(2-((4-hydroxybenzyl)(4-methoxyphenethyl)amino)-4-(4-methoxyphenyl)thiazol-5-yl)(4- methoxyphenyl)methanone; (2-((4-hydroxybenzyl)(4-methoxybenzyl)amino)-4-(4-methoxyphenyl)thiazol-5-yl)(4- methoxyphenyl)methanone;

(2-(butyl(4-hydroxybenzyl)amino)-4-phenylthiazol-5-yl)(phenyl)methanone; or

(2-amino-4-phenylthiazol-5-yl)(phenyl)methanone,

69. A method of treating cell proliferative diseases or conditions comprising administering to a patient an effective amount of a pharmaceutical composition of claim 63.

70. The method of claim 68, wherein the cell proliferative disease or condition is psoriasis, keloid scarring, or cancers of the breast, immune system, bone, nervous system, brain, blood, lymphatic system, or skin.

71. The method of claim 69, wherein the cell proliferative disease or condition is psoriasis, keloid scarring, or cancers of the breast, immune system, bone, nervous system, brain, blood, lymphatic system, or skin.

72. A method of treating conditions or diseases that involve TRAF6 comprising administering to a patient an effective amount of a compound of claim 55 or 61 or a compound selected from

(2-(butyl(4-hydroxybenzyl)amino)-4-phenylthiazol-5-yl)(phenyl)methanone; or

(2-amino-4-phenylthiazol-5-yl)(phenyl)methanone,

73. A method of treating conditions or diseases that involve TRAF6 comprising administering to a patient an effective amount of a pharmaceutical composition of claim 63.

74. The method of claim 72, wherein the conditions or diseases that involve TRAF6 is cancer, inflammation, adaptive immunity, innate immunity, bone metabolism, LPS-induced angiogenesis, osteoporosis, osteopinneal diseases, lymph node development, mammary gland development, skin development, or central nervous system development.

75. The method of claim 73, wherein the conditions or diseases that involve TRAF6 is cancer, inflammation, adaptive immunity, innate immunity, bone metabolism, LPS-induced angiogenesis, osteoporosis, osteopinneal diseases, lymph node development, mammary gland development, skin development, or central nervous system development.

76. A method of treating conditions or diseases that involve p27/SCF comprising administering to a patient an effective amount of a compound of claim 55 or 61 or a compound selected from

(2-(butyl(4-hydroxybenzyl)amino)-4-phenylthiazol-5-yl)(phenyl)methanone; or

(2-amino-4-phenylthiazol-5-yl)(phenyl)methanone,

N-CbenzoCdHl^dioxol-S-ylmethyO-S-ttert-butylamino^-^-β-^- (trifluoromethoxy)phenyl)ureido)phenyl)imidazo [ 1 ,2-a]pyridine-6-carboxamide; and

77. A method of treating conditions or diseases that involve p27/SCF comprising administering to a patient an effective amount of a pharmaceutical composition of claim 63.

78. The method of claim 76, wherein the conditions or diseases that involve p27/SCF is cancer, inflammation, adaptive immunity, innate immunity, bone metabolism, LPS-induced angiogenesis, osteoporosis, osteopinneal diseases, lymph node development, mammary gland development, skin development, or central nervous system development.

79. The method of claim 77, wherein the conditions or diseases that involve p27/SCF is cancer, inflammation, adaptive immunity, innate immunity, bone metabolism, LPS-induced angiogenesis, osteoporosis, osteopinneal diseases, lymph node development, mammary gland development, skin development, or central nervous system development.

80. A method of treating conditions or diseases that involve Radl8 comprising administering to a patient an effective amount of a compound of claim 55 or 61 or a compound selected from (2-((4-hydroxybenzyl)(4-methoxybenzyl)amino)-4-phenylthiazol-5- yl)(phenyl)methanone;

(2-(butyl(4-hydroxybenzyl)amino)-4-phenylthiazol-5-yl)(phenyl)methanone; or

(2-amino-4-phenylthiazol-5-yl)(phenyl)methanone,

81. A method of treating conditions or diseases that involve Radl 8 comprising administering to a patient an effective amount of a pharmaceutical composition of claim 63.

82. The method of claim 80, wherein the conditions or diseases that involve Radl 8 is cancer, inflammation, adaptive immunity, innate immunity, bone metabolism, LPS-induced angiogenesis, osteoporosis, osteopinneal diseases, lymph node development, mammary gland development, skin development, or central nervous system development.

83. The method of claim 81 , wherein the conditions or diseases that involve Radl 8 is cancer, inflammation, adaptive immunity, innate immunity, bone metabolism, LPS-induced angiogenesis, osteoporosis, osteopinneal diseases, lymph node development, mammary gland development, skin development, or central nervous system development.

84. A method of treating conditions or diseases that involve BMI-I comprising administering to a patient an effective amount of a compound of claim 55 or 61 or a compound selected from

(2-(butyl(4-hydroxybenzyl)amino)-4-phenylthiazol-5-yl)(phenyl)methanone; or

(2-amino-4-phenylthiazol-5-yl)(phenyl)methanone,

85. A method of treating conditions or diseases that involve BMI-I comprising administering to a patient an effective amount of a pharmaceutical composition of claim 63.

86. The method of claim 84, wherein the conditions or diseases that involve BMI-I is cancer, inflammation, adaptive immunity, innate immunity, bone metabolism, LPS-induced angiogenesis, osteoporosis, osteopinneal diseases, lymph node development, mammary gland development, skin development, or central nervous system development.

87. The method of claim 85, wherein the conditions or diseases that involve BMI-I is cancer, inflammation, adaptive immunity, innate immunity, bone metabolism, LPS-induced angiogenesis, osteoporosis, osteopinneal diseases, lymph node development, mammary gland development, skin development, or central nervous system development.

88. A pharmaceutical composition comprising a pharmaceutically acceptable carrier, diluent, or excipient and a compound of the formula,

wherein, n is 0, 1, 2, or 3;

R¹ is -CN or -C(O)NH(R⁵), wherein R⁵ is -H, Ci-C₆ alkyl, or Ci-C₆ haloalkyl;

R² is -H or -L-R⁷, wherein

L is -C(O)- or -S(O)₂-; and

R⁷ is Ci-C₆ alkyl, Ci-C₆ haloalkyl, C₃-Cg cycloalkyl, heterocyclyl, aryl, or heteroaryl, wherein R⁷ is optionally substituted with one to four R⁴ groups; R³ is -Ci-C₆ alkyl, -Ci-C₆ haloalkyl, aryl, -Ci-C₆ alkyl-aryl, wherein R³ is optionally substituted with one to four R⁴ groups; and R⁴ is halo, -OR⁶, -N(R⁶)₂, -S(R⁶), -S(O)₂R⁶, -S(O)₂N(R⁶)₂, -S(O)₂OR⁶,

-N(R⁶)S(O)₂R⁶,-OS(O)₂R⁶, -C(O)R⁶, -C(O)OR⁶, -C(O)N(R⁶)₂, -OC(O)R⁶, -OC(O)OR⁶, -OC(O)N(R⁶)₂, -N(R⁶)C(O)R⁶, -N(R⁶)C(O)OR⁶, -N(R⁶)C(O)N(R⁶)², -CN, -NO₂, -Ci-C₆ alkyl, -Ci -C₆ haloalkyl, -C₁-C₆ alkyl-OR⁶, or -C₁-C₆ alkyl-N(R⁶)₂, wherein each R⁶ is independently-H, Ci-C₆ alkyl, Ci-C₆ haloalkyl, C₃-Cg cycloalkyl, heterocyclyl, aryl, or heteroaryl.

89. The pharmaceutical composition of claim 88, wherein the compound is of the formula,

iition of claim 89, wherein

91. The pharmaceutical composition of claim 90, wherein R⁷ is aryl or heteroaryl, wherein R⁷ is optionally substituted with one to four R⁴ groups.

92. The pharmaceutical composition of claim 91, wherein R⁷ is phenyl optionally substituted with one or two R⁴ groups.

93. The pharmaceutical composition of claim 91, wherein R⁷ is furanyl, thienyl, pyrrolyl, thiazolyl, oxazolyl, isoxazoyl, isothiazoyl, imidazoyl, pyrazoyl, or triazolyl, wherein each R⁷ is optionally substituted with one to four R⁴ groups.

94. The pharmaceutical composition of claim 90, wherein R³ is -Ci-C₆ alkyl, phenyl, or benzyl, wherein R³ is optionally substituted with one or two R⁴ groups.

95. The pharmaceutical composition of claim 89, wherein the compound is

N-(l-butyl-3-cyano-lH-pyrrolo[2,3-b]quinoxalin-2-yl)-4-(N,N-dimethylsulfamoyl)benza mide;

5 -bromo-N-( 1 -butyl-3 -cyano- 1 H-pyrrolo [2,3 -b] quinoxalin-2-yl)furan-2-carboxamide;

N-(l-butyl-3-cyano-lH-pyrrolo[2,3-b]quinoxalin-2-yl)-4-fluorobenzamide;

N-(3-cyano-l-pentyl-lH-pyrrolo[2,3-b]quinoxalin-2-yl)-4-fluorobenzamide;

N-(l-butyl-3-cyano-lH-pyrrolo[2,3-b]quinoxalin-2-yl)benzamide;

N-(l-butyl-3-cyano-lH-pyrrolo[2,3-b]quinoxalin-2-yl)-4-chlorobenzamide;

N-(3 -cyano- 1 -(2 -methoxy ethyl)- 1 H-pyrrolo [2,3 -b] quinoxalin-2-yl)benzamide;

N-(l-butyl-3-cyano-lH-pyrrolo[2,3-b]quinoxalin-2-yl)thiophene-2-carboxamide; N-(l-allyl-3-cyano-lH-pyrrolo[2,3-b]quinoxalin-2-yl)benzamide; N-(l-sec-butyl-3-cyano-lH-pyrrolo[2,3-b]quinoxalin-2-yl)-3,4,5-trimethoxybenzamide; 5 -bromo-N-(3 -cyano- 1 -(4-methoxyphenyl)- 1 H-pyrrolo [2,3 -b] quinoxalin-2-yl)furan-2-car boxamide;

N-(l-benzyl-3-cyano-lH-pyrrolo[2,3-b]quinoxalin-2-yl)furan-2-carboxamide; 4-chloro-N-(3 -cyano- 1 -phenyl- lH-pyrrolo[2,3-b]quinoxalin-2-yl)benzamide; N-(3-cyano-l -(4-methoxyphenyl)- lH-pyrrolo[2,3-b]quinoxalin-2-yl)benzamide; 5-bromo-N-(l-(4-bromophenyl)-3-cyano-lH-pyrrolo[2,3-b]quinoxalin-2-yl)furan-2-carbo xamide;

5-bromo-N-(3-cyano-l-(3-hydroxypropyl)-lH-pyrrolo[2,3-b]quinoxalin-2-yl)furan-2-carb oxamide; or 5-bromo-N-(l-sec-butyl-3-cyano-lH-pyrrolo[2,3-b]quinoxalin-2-yl)furan-2-carboxamide.

96. A method of inhibiting ubiquitination in a cell comprising contacting the cell in which inhibition of ubiquitination is desired with a pharmaceutical composition of claim 88.

97. The method of claim 96, wherein ubiquitin ligase activity of p27/SCF, TRAF6, Radl8, or BMI-I is inhibited.

98. A method of treating cell proliferative diseases or conditions comprising administering to a patient an effective amount of a pharmaceutical composition of claim 88.

99. The method of claim 98, wherein the cell proliferative disease or condition is psoriasis, keloid scarring, or cancers of the breast, immune system, bone, nervous system, brain, blood, lymphatic system, or skin.

100. A method of treating conditions or diseases that involve TRAF6 comprising administering to a patient an effective amount of a pharmaceutical composition of claim 88.

101. The method of claim 100, wherein the conditions or diseases that involve TRAF6 is cancer, inflammation, adaptive immunity, innate immunity, bone metabolism, LPS-induced angiogenesis, osteoporosis, osteopinneal diseases, lymph node development, mammary gland development, skin development, or central nervous system development.

102. A method of treating conditions or diseases that involve p27/SCF comprising administering to a patient an effective amount of a pharmaceutical composition of claim 88.

103. The method of claim 102, wherein the conditions or diseases that involve p27/SCF is cancer, inflammation, adaptive immunity, innate immunity, bone metabolism, LPS-induced angiogenesis, osteoporosis, osteopinneal diseases, lymph node development, mammary gland development, skin development, or central nervous system development.

104. A method of treating conditions or diseases that involve Radl8 comprising administering to a patient an effective amount of a pharmaceutical composition of claim 88.

105. The method of claim 104, wherein the conditions or diseases that involve Radl8 is cancer, inflammation, adaptive immunity, innate immunity, bone metabolism, LPS-induced angiogenesis, osteoporosis, osteopinneal diseases, lymph node development, mammary gland development, skin development, or central nervous system development.

106. A method of treating conditions or diseases that involve BMI-I comprising administering to a patient an effective amount of a pharmaceutical composition of claim 88.

107. The method of claim 106, wherein the conditions or diseases that involve BMI-I is cancer, inflammation, adaptive immunity, innate immunity, bone metabolism, LPS-induced angiogenesis, osteoporosis, osteopinneal diseases, lymph node development, mammary gland development, skin development, or central nervous system development.