# Patent

## Title
Heteroaryl-substituted bicyclic Smac mimetics and the uses thereof

## Abstract

The invention relates to heteroaryl-substituted bicyclic mimetics of Smac which function as inhibitors of Inhibitor of Apoptosis Proteins. The invention also relates to the use of these mimetics for inducing apoptotic cell death and for sensitizing cells to inducers of apoptosis.

## Background
CROSS REFERENCE TO RELATED APPLICATIONS
The present application is a U.S. 371 National Stage Entry of pending International Patent Application No. PCT/US2009/040312, International Filing Date Apr. 13, 2009, which claims priority to expired U.S. Provisional Patent Application No. 61/044,330 filed Apr. 11, 2008, and expired U.S. Provisional Patent Application No. 61/106,887 filed Oct. 20, 2008, both of which are herein incorporated by reference in their entireties.


STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT
This invention was made with government support under R01CA109025 awarded by the National Institutes of Health. The government has certain rights in the invention.


FIELD OF THE INVENTION
This invention is in the field of medicinal chemistry. In particular, the invention relates to heteroaryl-substituted bicyclic Smac mimetics of the N-terminal sequence of Smac which function as inhibitors of Inhibitor of Apoptosis Proteins. The invention also relates to the use of these mimetics for inducing or sensitizing cells to the induction of apoptotic cell death.
RELATED ART
The aggressive cancer cell phenotype is the result of a variety of genetic and epigenetic alterations leading to deregulation of intracellular signaling pathways (Ponder, Nature 411:336 (2001)). The commonality for all cancer cells, however, is their failure to execute an apoptotic program, and lack of appropriate apoptosis due to defects in the normal apoptosis machinery is a hallmark of cancer (Lowe et al., Carcinogenesis 21:485 (2000)). Most current cancer therapies, including chemotherapeutic agents, radiation, and immunotherapy, work by indirectly inducing apoptosis in cancer cells. The inability of cancer cells to execute an apoptotic program due to defects in the normal apoptotic machinery is thus often associated with an increase in resistance to chemotherapy, radiation, or immunotherapy-induced apoptosis. Primary or acquired resistance of human cancer of different origins to current treatment protocols due to apoptosis defects is a major problem in current cancer therapy (Lowe et al., Carcinogenesis 21:485 (2000); Nicholson, Nature 407:810 (2000)). Accordingly, current and future efforts towards designing and developing new molecular target-specific anticancer therapies to improve survival and quality of life of cancer patients must include strategies that specifically target cancer cell resistance to apoptosis. In this regard, targeting crucial negative regulators that play a central role in directly inhibiting apoptosis in cancer cells represents a highly promising therapeutic strategy for new anticancer drug design.
Two classes of central negative regulators of apoptosis have been identified. The first class of regulators is the Bcl-2 family of proteins, as exemplified by two potent anti-apoptotic molecules, Bcl-2 and Bcl-XL proteins (Adams et al., Science 281:1322 (1998); Reed, Adv. Pharmacol. 41:501 (1997); Reed et al., J. Cell. Biochem. 60:23 (1996)). Therapeutic strategies for targeting Bcl-2 and Bcl-XL in cancer to restore cancer cell sensitivity and overcome resistance of cancer cells to apoptosis have been extensively reviewed (Adams et al., Science 281:1322 (1998); Reed, Adv. Pharmacol. 41:501 (1997); Reed et al., J. Cell. Biochem. 60:23 (1996)). Several laboratories are interested in designing small molecule inhibitors of Bcl-2 and Bcl-XL.
The second class of central negative regulators of apoptosis is the inhibitor of apoptosis proteins (IAPs) (Deveraux et al., Genes Dev. 13:239 (1999); Salvesen et al., Nat. Rev. Mol. Cell. Biol. 3:401 (2002)). This class includes proteins such as XIAP, cIAP-1, cIAP-2, ML-IAP, HIAP, KIAP, TSIAP, NAIP, survivin, livin, ILP-2, apollon, and BRUCE. IAP proteins potently suppress apoptosis induced by a large variety of apoptotic stimuli, including chemotherapeutic agents, radiation, and immunotherapy in cancer cells.
X-linked IAP(XIAP) is the most potent inhibitor in suppressing apoptosis among all of the IAP members (Holcik et al., Apoptosis 6:253 (2001); LaCasse et al., Oncogene 17:3247 (1998); Takahashi et al., J. Biol. Chem. 273:7787 (1998); Deveraux et al., Nature 388:300 (1997); Sun et al., Nature 401:818 (1999); Deveraux et al., EMBO J. 18:5242 (1999); Asselin et al., Cancer Res. 61:1862 (2001)). XIAP plays a key role in the negative regulation of apoptosis in both the death receptor-mediated and the mitochondria-mediated pathways. XIAP functions as a potent endogenous apoptosis inhibitor by directly binding and potently inhibiting three members of the caspase family of enzymes, caspase-3, -7, and -9 (Takahashi et al., J. Biol. Chem. 273:7787 (1998); Deveraux et al., Nature 388:300 (1997); Sun et al., Nature 401:818 (1999); Deveraux et al., EMBO J. 18:5242 (1999); Asselin et al., Cancer Res. 61:1862 (2001); Riedl et al., Cell 104:791 (2001); Chai et al., Cell 104:769 (2001); Huang et al., Cell 104:781 (2001)). XIAP contains three baculovirus inhibitor of apoptosis repeat (BIR) domains as well as a C-terminal RING finger. The third BIR domain (BIR3) selectively targets caspase-9, the initiator caspase in the mitochondrial pathway, whereas the linker region between BIR1 and BIR2 inhibits both caspase-3 and caspase-7 (Salvesen et al., Nat. Rev. Mol. Cell. Biol. 3:401 (2002)). While binding to XIAP prevents the activation of all three caspases, it is apparent that the interaction with caspase-9 is the most critical for its inhibition of apoptosis (Ekert et al., J. Cell Biol. 152:483 (2001); Srinivasula et al., Nature 410:112 (2001)). Because XIAP blocks apoptosis at the down-stream effector phase, a point where multiple signaling pathways converge, strategies targeting XIAP may prove to be especially effective to overcome resistance of cancer cells to apoptosis (Fulda et al., Nature Med. 8:808 (2002); Arnt et al., J. Biol. Chem. 277:44236 (2002)).
Although the precise role of XIAP in each type of cancer is far from completely understood, evidence is mounting to indicate that XIAP is widely overexpressed in many types of cancer and may play an important role in the resistance of cancer cells to a variety of current therapeutic agents (Holcik et al., Apoptosis 6:253 (2001); LaCasse et al., Oncogene 17:3247 (1998)).
XIAP protein was found to be expressed in most of the NCI 60 human cancer cell lines (Tamm et al., Clin. Cancer Res. 6:1796 (2000)). Analysis of tumor samples in 78 previously untreated patients showed that those with lower levels of XIAP had significantly longer survival (Tamm et al., Clin. Cancer Res. 6:1796 (2000)). XIAP was found to be expressed in human malignant glioma (Wagenknecht et al., Cell Death Differ. 6:370 (1999); Fulda et al., Nature Med. 8:808 (2002)). XIAP was found to be expressed in human prostate cancer cells and blocks Apo2 ligand/tumor necrosis factor-related apoptosis inducing ligand-mediated apoptosis of prostate cancer cells in the presence of mitochondrial activation (McEleny et al., Prostate 51:133 (2002); Ng et al., Mol. Cancer. Ther. 1:1051 (2002)). XIAP is overexpressed in non-small cell lung cancer (NSCLC) in patients and has been implicated in pathogenesis of NSCLC (Hofmann et al., J. Cancer Res. Clin. Oncol. 128:554 (2002)). Expression of XIAP and lack of down-regulation of XIAP upon treatment with cisplatin have been implicated in cisplatin resistance of human ovarian cancer (Li et al., Endocrinology 142:370 (2001); Cheng et al., Drug Resist. Update 5:131 (2002)). Taken together, these data suggest that XIAP may play an important role in resistance of several human cancers to current therapeutic agents.
Integrity of the blood vessel wall is essential for vascular homeostasis and organ function. A dynamic balance between endothelial cell survival and apoptosis contributes to this integrity during vascular development and pathological angiogenesis. It has been shown that cIAP-1 is essential for maintaining endothelial cell survival and blood vessel homeostasis during vascular development (Santoro et al., Nature Genetics 39:1397 (2007). As such, cIAP-1 may play an important role in the control of angiogenesis and blood vessel homeostasis during embryogenesis, regeneration and tumorigenesis.
Apoptosis is not a single process, rather, it is involved with a number of different, sometimes interconnected, signaling pathways leading to cell degradation. The pathways involved in a particular form of apoptosis depend on many factors, such as the insult or insults that initiate the process. Other factors include the activation or overactivation of specific receptors, such as the activation of “death” receptors by tumor necrosis factor alpha (TNFα), tumor necrosis factor-related apoptosis-inducing ligand (TRAIL or Apo2L), or FAS ligand. Another determining factor is the type of cell which is involved, since different signaling pathways are shown for so called type I and type II cells after Fas or TNFα receptor activation.
TRAIL (Apo2L) has been shown to be a selective and potent inducer of apoptosis in cancer cells (but not normal cells) upon binding to either of two pro-apoptotic TRAIL receptors, TRAIL-R1 (or DR4) (Pan et al., Science 276:111 (1997)) or TRAIL-R2 (KILLER, or DR5) (Wu et al., Nat. Genet. 17:141-143 (1997); Pan et al., Science 277:815 (1997); Walczak et al., EMBO J. 16:5386 (1997)). Activation of the pro-apoptotic death receptors by TRAIL induces the formation of death inducing signaling complex (DISC), which consists of receptor FADD as an adaptor (Kischkel et al., Immunity 12:611 (2000); Kuang et al., J. Biol. Chem. 275:25065 (2000)), and caspase-8 as an initiator caspase. Once DISC is formed, caspase-8 is auto-processed and activated by induced proximity (Medema et al., EMBO J. 16:2794 (1997); Muzio et al., J. Biol. Chem. 273:2926 (1998)).
TRAIL has generated significant interest as a potential cancer therapeutic (French et al., Nat. Med. 5:146 (1999)) because of its selective targeting of cancer cells, whereas most normal cells appear to be resistant to TRAIL (Ashkenazi et al., Science 281:1305 (1998); Walczak et al., Nat. Med. 5:157 (1999)). Systemic administration of TRAIL has proven to be safe and effective at killing breast or colon xenografted tumors and prolonging survival in mice (Walczak et al., Nat. Med. 5:157 (1999)). Although TRAIL can specifically kill many types of cancer cells, many others display TRAIL-resistance (Kim et al., Clin. Cancer Res. 6:335 (2000); Zhang et al., Cancer Res. 59:2747 (1999)). In addition, cancer cells have been killed by application of antibodies (monoclonal or polyclonal) that specifically recognize either TRAIL-R1 or TRAIL-R2.
Numerous mechanisms have been identified as potential factors responsible for TRAIL-resistance. Such mechanisms exist at a number of levels, including at the receptor level, mitochondria level, post-mitochondria level, and at the DISC level. For example, loss of caspase-8 expression (Teitz et al., Nat. Med. 6:529 (2000); Griffith et al., J. Immunol. 161:2833 (1998)), or high expression of the cellular FLICE inhibitor protein (cFLIP) (Kim et al., Clin. Cancer Res. 6:335 (2000); Zhang et al., Cancer Res. 59:2747 1999; Kataoka et al., J. Immunol. 161:3936 (1998)) make cancer cells resistant to TRAIL. Yeh et al. have shown that cFLIP-deficient embryonic mouse fibroblasts are particularly sensitive to receptor-mediated apoptosis (Yeh et al., Immunity 12:533 (2000)). Several splice variants of cFLIP are known, including a short splice variant, cFLIP-S, and a longer splice variant, cFLIP-L. It has been shown that cFLIP-deficient embryonic mouse fibroblasts become resistant to TRAIL-induced apoptosis as a result of retroviral-mediated transduction of cFLIP-S (Bin et al., FEBS Lett. 510:37 (2002)).
Although TRAIL represents a potentially promising candidate for tumor-selective death receptor activation (i.e., it induces apoptosis preferentially in tumor cells but not in normal tissues), many cancer cells are resistant to apoptosis-inducing drugs, as discussed above. As a result, treatment with such drugs often requires co-treatment with irradiation and/or cytotoxic chemicals to achieve a therapeutic effect. However, both radiation and chemotherapy have significant side effects, and are generally avoided if possible.
Thus, a need exists for an agent that can selectively and efficiently sensitize tumor cells to selective, apoptosis-inducing drugs such as TRAIL or TRAIL receptor antibodies, without also sensitizing surrounding normal cells. Such an agent would also be useful for reducing or preventing the drug resistance commonly associated with the use of receptor-mediated apoptotic cancer drugs, thus improving their effectiveness and eliminating the need for combination therapies.
Recently, Smac/DIABLO (second mitochondria-derived activator of caspases) was identified as a protein released from mitochondria into the cytosol in response to apoptotic stimuli (Budihardjo et al., Annu. Rev. Cell Dev. Biol. 15:269 (1999); Du et al., Cell 102:33 (2000)). Smac is synthesized with an N-terminal mitochondrial targeting sequence that is proteolytically removed during maturation to the mature polypeptide. Smac was shown to directly interact with XIAP and other IAPs and to disrupt their binding to caspases and facilitate caspase activation. Smac is a potent endogenous inhibitor of XIAP.
High resolution, experimental three-dimensional (3D) structures of the BIR3 domain of XIAP in complex with Smac protein and peptide have recently been determined (Sun et al., J. Biol. Chem. 275:36152 (2000); Wu et al., Nature 408:1008 (2000)) (FIG. 1). The N-terminal tetrapeptide of Smac (Ala-Val-Pro-Ile, or AVPI (SEQ ID NO:1)) recognizes a surface groove on the BIR3 domain of XIAP through several hydrogen-bonding interactions and van der Waals contacts. The interaction between BIR3 and caspase-9 has also been shown to involve four residues (Ala-Thr-Pro-Phe, or ATPF (SEQ ID NO:2)) on the amino terminus of the small subunit of caspase-9 to the same surface groove on the BIR3 domain. Several recent studies have convincingly demonstrated that Smac promotes the catalytic activity of caspase-9 by competing with caspase-9 for the same binding groove on the surface of the BIR3 domain (Ekert et al., J. Cell Biol. 152:483 (2001); Srinivasula et al., Nature 410:112 (2001)).
Unlike most protein-protein interactions, the Smac-XIAP interaction is mediated by only four amino acid residues on the Smac protein and a well-defined surface groove on the BIR3 domain of XIAP. The Kd value of Smac peptide AVPI (SEQ ID NO:1) to XIAP BIR3 (Kd=0.4 μM) is essentially the same as the mature Smac protein (Kd=0.42 μM). This well-defined interaction site is ideal for the design of non-peptide, drug-like small molecules that mimic the binding of Smac to XIAP.
A cell permeable Smac peptide, which consists of the first four amino acid residues (AVPI (SEQ ID NO:1)) of the N-terminus of Smac tethered to a carrier peptide to facilitate intracellular delivery, was recently shown to sensitize various tumor cells in vitro and malignant glioma cells in vivo to apoptosis induced by death receptor ligation or cytotoxic drugs (Fulda et al., Nature Med. 8:808 (2002)). Importantly, this Smac peptide strongly enhanced the anti-tumor activity of Apo2L/TRAIL in an intracranial malignant glioma xenograft model in vivo. Complete eradication of established tumors and survival of mice was only achieved upon combined treatment with Smac peptides and Apo2L/TRAIL. Of significance, Smac peptide does not have detectable toxicity to normal brain tissue.
A second recent independent study also showed that peptides consisting of the first four to eight amino acid residues of the N-terminus of Smac tethered to a different carrier peptide enhanced the induction of apoptosis and the long term anti-proliferative effects of diverse chemotherapeutic drugs, including paclitaxel, etoposide, SN-38, and doxorubicin in MCF-7 and other human breast cancer cell lines (Arnt et al., J. Biol. Chem. 277:44236 (2002). This study conclusively showed that XIAP and cIAP-1 are the primary molecular targets for these peptides in cells.
A third study showed that a Smac peptide of the first seven N-terminal residues tethered to polyarginine restored the apoptosome activity and reversed the apoptosis resistance in non-small cell lung cancer H460 cells (Yang et al., Cancer Res. 63:831 (2003)). XIAP was shown to be responsible for the defect in apoptosome activity and suppression of caspase activity in H460 cells. When used in combination with chemotherapy, the cell-permeable Smac peptide regressed tumor growth in vivo with little murine toxicity. Taken together, these recent independent studies strongly suggest that a potent, stable, cell-permeable Smac mimetic may have great therapeutic potential for the treatment of human breast cancer and other types of cancer.
Peptide-based inhibitors are useful tools to elucidate the anti-apoptotic function of IAPs and the role of IAPs in response of cancer cells to chemotherapeutic agents. But peptide-based inhibitors in general have intrinsic limitations as potentially useful therapeutic agents. These limitations include their poor cell-permeability and poor in vivo stability. Indeed, in these three published studies using Smac-based peptide inhibitors, the peptides had to be fused to carrier peptides to make them relatively cell-permeable.
To overcome the intrinsic limitations of peptide-based inhibitors, the present invention provides conformationally constrained Smac mimetics having heteroaromatic substitution on the bicyclic scaffold.
SUMMARY OF THE INVENTION
It is generally accepted that the inability of cancer cells or their supporting cells to undergo apoptosis in response to genetic lesions or exposure to inducers of apoptosis (such as anticancer agents and radiation) is a major factor in the onset and progression of cancer. The induction of apoptosis in cancer cells or their supporting cells (e.g., neovascular cells in the tumor vasculature) is thought to be a universal mechanism of action for virtually all of the effective cancer therapeutic drugs or radiation therapies on the market or in practice today. One reason for the inability of a cell to undergo apoptosis is increased expression and accumulation of IAPs.
The present invention contemplates that exposure of animals suffering from cancer or other hyperproliferative disorders or diseases associated with dysregulation of apoptosis to therapeutically effective amounts of drug(s) (e.g., small molecules) that inhibit the function(s) of IAPs will kill the diseased cells or supporting cells outright (those cells whose continued survival is dependent on the overactivity or overexpression of IAPs) and/or render such cells as a population more susceptible to the cell death-inducing activity of cancer therapeutic drugs or radiation therapies. The present invention contemplates that inhibitors of IAPs satisfy an unmet need for the treatment of multiple cancer types, either when administered as monotherapy to induce apoptosis in cancer cells dependent on IAP function, or when administered in a temporal relationship with other cell death-inducing cancer therapeutic drugs or radiation therapies so as to render a greater proportion of the cancer cells or supportive cells susceptible to executing the apoptosis program compared to the corresponding proportion of cells in an animal treated only with the cancer therapeutic drug or radiation therapy alone.
The present invention also contemplates that treatment of animals suffering from endothelial cell-associated diseases (e.g., tumor angiogenesis, retinopathies and atherosclerosis) with therapeutically effective amounts of drug(s) (e.g., small molecules) that inhibit the function(s) of IAPs (e.g., cIAP-1) may prevent or inhibit angiogenesis and disrupt blood vessel homeostasis during vascular development in pathological conditions. Particular disorders that may be treated with the compounds of the invention include macular degeneration, rheumatoid arthritis, psoriasis, diabetic retinopathy, retinopathy of prematurity, corneal graft rejection, neovascular glaucoma, retrolental fibroplasia, rubeosis, Osler-Webber Syndrome, myocardial angiogenesis, plaque neovascularization, telangiectasia, hemophiliac joints, angiofibroma, wound granulation, intestinal adhesions, atherosclerosis, scleroderma and hypertrophic scars.
Applicants have found that certain Smac mimetics having heteroaryl substitution on the bicyclic scaffold display unexpected in vitro potency in cancer cell lines. Thus, the compounds of the invention are expected to be useful for the treatment of a wide variety of diseases responsive to the induction of apoptotic cell death.
In certain embodiments of the invention, combination treatment of animals with a therapeutically effective amount of a compound of the present invention and a course of an anticancer agent or radiation is expected to produce a greater tumor response and clinical benefit in such animals compared to those treated with the compound or anticancer drugs/radiation alone. Put another way, because it is believed the compounds of the present invention lower the apoptotic threshold of all cells that express IAPs, the proportion of cells that successfully execute the apoptosis program in response to the apoptosis inducing activity of anticancer drugs/radiation is increased. Alternatively, the compounds of the present invention are expected to allow administration of a lower, and therefore less toxic and more tolerable, dose of an anticancer agent and/or radiation to produce the same tumor response/clinical benefit as the conventional dose of the anticancer agent/radiation alone. Since the doses for all approved anticancer drugs and radiation treatments are known, the present invention contemplates the various combinations of them with the compounds of the present invention. Also, since the compounds of the present invention act at least in part by inhibiting IAPs, the exposure of cancer cells and supporting cells to therapeutically effective amounts of the compounds can be temporally linked to coincide with the attempts of cells to execute the apoptosis program in response to the anticancer agent or radiation therapy. Thus, in some embodiments, administering the compositions of the present invention in connection with certain temporal relationships, is expected to provide especially efficacious therapeutic practices.
The present invention relates to Smac mimetics that are useful for inhibiting the activity of IAP proteins and inter alia increasing the sensitivity of cells to inducers of apoptosis. In one particular embodiment, the Smac mimetics are compounds of Formula I:




wherein:

A1 and A2 are independently selected from the group consisting of hydrogen and optionally substituted alkyl, wherein A2 is absent when V is O;

V is selected from the group consisting of N, CH and O;

W is selected from the group consisting of CH and N;

X is selected from the group consisting of hydrogen, optionally substituted alkyl and aralkyl;

Y is selected from the group consisting of CON(R1), N(R1)CO, C(O)O, OC(O), (CH2)1-3, wherein one or more CH2 groups can be replaced by O, S, or NR1, optionally substituted aryl and optionally substituted heteroaryl;

Z is (CR2aR2b)r;

D is (CR3aR3b)n—U—(CR4aR4b)m;

U is selected from the group consisting of CR5aR5b and NR6;

J is (CR7aR7b)p-L-(CR8aR8b)q;

T is optionally substituted heteroaryl;

n, m, p and q are independently selected from the group consisting of 0-5;

r is 0-3;

R1 is selected from the group consisting of hydrogen, optionally substituted alkyl, aralkyl, optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heteroaryl and optionally substituted heterocyclo;

each R2a, R2b, R3a, R3b, R4a, R4b, R5a, R5b, R7a, R7b, R8a, R8b are independently selected from the group consisting of hydrogen, optionally substituted alkyl, aralkyl, optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heteroaryl and optionally substituted heterocyclo;

R6 is selected from the group consisting of hydrogen, optionally substituted alkyl, aralkyl, optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heteroaryl and optionally substituted heterocyclo and COR9;

L is selected from the group consisting of O, S, NR1, NCOR9, CR7aR7b, C═O, C═S and C═NR1; and

R9 is selected from the group consisting of hydrogen, optionally substituted alkyl, aralkyl, optionally substituted cycloalkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted aryl, optionally substituted heteroaryl and optionally substituted heterocyclo;

or pharmaceutically acceptable salt or prodrug thereof.

In another particular embodiment, Smac mimetics are compounds of Formula II:




wherein A1, A2, V, Z, W, X, Y, D, J and T have the meanings as described above for Formula I; or a pharmaceutically acceptable salt or prodrug thereof.

In another particular embodiment, Smac mimetics are compounds of Formula III:




wherein A1, A2, V, Z, W, X, Y, U, and T have the meanings as described above for Formula I, and m is 1 or 2; or a pharmaceutically acceptable salt or prodrug thereof.

In another particular embodiment, Smac mimetics are compounds of Formula IV:




wherein A1, A2, X, U, and T have the meanings as described above for Formula I, and m is 1 or 2; or a pharmaceutically acceptable salt or prodrug thereof.

In another particular embodiment, Smac mimetics are compounds of Formula V:




wherein A1, A2, X, U and T have the meanings as described above for Formula I, and m is 1 or 2; or a pharmaceutically acceptable salt or prodrug thereof.

In another particular embodiment, Smac mimetics are compounds of Formula VI:




wherein A1 and X are optionally substituted alkyl and T is optionally substituted heteroaryl; or a pharmaceutically acceptable salt or prodrug thereof.

In another particular embodiment, Smac mimetics are compounds of Formula VII:




wherein A1 and X are optionally substituted alkyl, R9 is optionally substituted alkyl or aralkyl, and T is optionally substituted heteroaryl; or a pharmaceutically acceptable salt or prodrug thereof.

In another particular embodiment, Smac mimetics are compounds of Formulae I-VII wherein T is




wherein Q is O, S or NR12, R12 is hydrogen, optionally substituted alkyl, aralkyl, optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heteroaryl or optionally substituted heterocyclo, R10a, R10b, R11a, R11b, R11c and R11d are independently selected from the group consisting of hydrogen, optionally substituted alkyl, haloalkyl, aralkyl, optionally substituted cycloalkyl, optionally substituted cycloalkenyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocyclo, halo, nitro, cyano, hydroxy, amino, alkoxy, aryloxy, arylalkyloxy, alkylthio, carboxamido and sulfonamido, Z1, Z2, and Z3 are independently CR11e or N, wherein at least one of Z1, Z2, and Z3 is CR11e and at least one of Z1, Z2, and Z3 is N, and R11e is selected from the group consisting of hydrogen, optionally substituted alkyl, haloalkyl, aralkyl, optionally substituted cycloalkyl, optionally substituted cycloalkenyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocyclo, halo, nitro, cyano, hydroxy, amino, alkoxy, aryloxy, arylalkyloxy, alkylthio, carboxamido and sulfonamido; or a pharmaceutically acceptable salt or prodrug thereof.

In another particular embodiment, Smac mimetics are compounds of Formulae I-VII wherein T is




wherein Q, Z1, Z2, Z3, R10a, R10b, R11a, R11b, R11c and R11d have the meanings described above; or pharmaceutically acceptable salt or prodrug thereof.

In another particular embodiment, Smac mimetics are compounds of Formula VIII:




wherein A1 and X are optionally substituted alkyl, m is 1 or 2, Q is O, S or NR12, R12 is hydrogen, optionally substituted alkyl, aralkyl, optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heteroaryl or optionally substituted heterocyclo, and R10a and R10b are independently selected from the group consisting of hydrogen, optionally substituted alkyl, haloalkyl, aralkyl, optionally substituted cycloalkyl, optionally substituted cycloalkenyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocyclo, halo, nitro, cyano, hydroxy, amino, alkoxy, aryloxy, arylalkyloxy, alkylthio, carboxamido and sulfonamido; or a pharmaceutically acceptable salt or prodrug thereof.

In another particular embodiment, Smac mimetics are compounds of Formula IX:




wherein A1 and X are optionally substituted alkyl, m is 1 or 2, Q is O, S or NR12, R12 is hydrogen, optionally substituted alkyl, aralkyl, optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heteroaryl or optionally substituted heterocyclo, and R11a, R11b, R11c and R11d are independently selected from the group consisting of hydrogen, optionally substituted alkyl, haloalkyl, aralkyl, optionally substituted cycloalkyl, optionally substituted cycloalkenyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocyclo, halo, nitro, cyano, hydroxy, amino, alkoxy, aryloxy, arylalkyloxy, alkylthio, carboxamido and sulfonamido; or a pharmaceutically acceptable salt or prodrug thereof.

In another particular embodiment, Smac mimetics are compounds of Formula X:




wherein A1 and X are optionally substituted alkyl, R9 is optionally substituted alkyl or aralkyl, m is 1 or 2, Q is O, S or NR12, R12 is hydrogen, optionally substituted alkyl, aralkyl, optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heteroaryl or optionally substituted heterocyclo, and R10a and R10b are independently selected from the group consisting of hydrogen, optionally substituted alkyl, haloalkyl, aralkyl, optionally substituted cycloalkyl, optionally substituted cycloalkenyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocyclo, halo, nitro, cyano, hydroxy, amino, alkoxy, aryloxy, arylalkyloxy, alkylthio, carboxamido and sulfonamido; or a pharmaceutically acceptable salt or prodrug thereof.

In another particular embodiment, Smac mimetics are compounds of Formula XI:




wherein A1 and X are optionally substituted alkyl, R9 is optionally substituted alkyl or aralkyl, m is 1 or 2, Q is O, S or NR12, R12 is hydrogen, optionally substituted alkyl, aralkyl, optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heteroaryl or optionally substituted heterocyclo, and R11a, R11b, R11c and R11d are independently selected from the group consisting of hydrogen, optionally substituted alkyl, haloalkyl, aralkyl, optionally substituted cycloalkyl, optionally substituted cycloalkenyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocyclo, halo, nitro, cyano, hydroxy, amino, alkoxy, aryloxy, arylalkyloxy, alkylthio, carboxamido and sulfonamido; or a pharmaceutically acceptable salt or prodrug thereof.

The invention relates to compounds represented by Formulae I-XIa which are inhibitors of IAP proteins. The invention relates to the use of the compounds of the invention to induce apoptosis in cells and inhibit angiogenesis. The invention also relates to the use of the compounds of the invention for sensitizing cells to inducers of apoptosis. The compounds are useful for the treatment, amelioration, or prevention of disorders responsive to induction of apoptotic cell death, e.g., disorders characterized by dysregulation of apoptosis, including hyperproliferative diseases such as cancer. In certain embodiments, the compounds can be used to treat, ameliorate, or prevent cancer that is characterized by resistance to cancer therapies (e.g., those which are chemoresistant, radiation resistant, hormone resistant, and the like). In other embodiments, the compounds can be used to treat hyperproliferative diseases characterized by overexpression of IAPs. In other embodiments, the compounds can be used as a method of preventing or inhibiting angiogenesis in animals in need thereof. The present invention provides pharmaceutical compositions comprising compounds of Formulae I-XIa in a therapeutically effective amount to induce apoptosis in cells or to sensitize cells to inducers of apoptosis.
The invention further provides kits comprising a compound of Formula I and instructions for administering the compound to an animal. The kits may optionally contain other therapeutic agents, e.g., anticancer agents or apoptosis-modulating agents.
The present invention also provides a process for preparing a compound of Formula XII




comprising:

a) condensing a compound of Formula XIII





with ammonia to give a compound of Formula XIV





b) converting a compound of Formula XIV to a compound of Formula XV





c) condensing a compound of Formula XV with a compound of Formula XVI





wherein L2 is a leaving group, to give a compound of Formula XVII,





and b) cyclizing a compound of Formula XVII, to give a compound of Formula XII, wherein:

R13 is selected from the group consisting of N(H)P1 and





P1 is an amine protecting group;

A1 and A2 are independently selected from the group consisting of hydrogen and optionally substituted alkyl, wherein A2 is absent when V is O;

V is selected from the group consisting of N, CH and O;

W is selected from the group consisting of CH and N;

X is selected from the group consisting of hydrogen, optionally substituted alkyl and aralkyl;

Y is selected from the group consisting of CON(R1), N(R1)CO, C(O)O, OC(O), (CH2)1-3, wherein one or more CH2 groups can be replaced by O, S, or NR1, optionally substituted aryl and optionally substituted heteroaryl;

Z is (CR2aR2b)r;

U is selected from the group consisting of CR5aR5b and NR6;

m is 1 or 2;

r is 0-3;

R1 is selected from the group consisting of hydrogen, optionally substituted alkyl, aralkyl, optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heteroaryl and optionally substituted heterocyclo;

each R2a, R2b, R5a and R5b are independently selected from the group consisting of hydrogen, optionally substituted alkyl, aralkyl, optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heteroaryl and optionally substituted heterocyclo;

R6 is selected from the group consisting of hydrogen, optionally substituted alkyl, aralkyl, optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heteroaryl and optionally substituted heterocyclo and COR9;

R9 is selected from the group consisting of hydrogen, optionally substituted alkyl, aralkyl, optionally substituted cycloalkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted aryl, optionally substituted heteroaryl and optionally substituted heterocyclo; and

R10a and R10b are independently selected from the group consisting of hydrogen, optionally substituted alkyl, haloalkyl, aralkyl, optionally substituted cycloalkyl, optionally substituted cycloalkenyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted aryl, optionally substituted heteroaryl and optionally substituted heterocyclo.
The present invention also provides a process for preparing a compound of Formula XVIII




comprising:

a) condensing a compound of Formula XIII





with a compound of Formula XIX





to give a compound of Formula XX





and b) cyclizing a compound of Formula XX, to give a compound of Formula XVIII, wherein:

R13 is selected from the group consisting of N(H)P1 and





P1 is an amine protecting group;

A1 and A2 are independently selected from the group consisting of hydrogen and optionally substituted alkyl, wherein A2 is absent when V is O;

V is selected from the group consisting of N, CH and O;

W is selected from the group consisting of CH and N;

X is selected from the group consisting of hydrogen, optionally substituted alkyl and aralkyl;

Y is selected from the group consisting of CON(R1), N(R1)CO, C(O)O, OC(O), (CH2)1-3, wherein one or more CH2 groups can be replaced by O, S, or NR1, optionally substituted aryl and optionally substituted heteroaryl;

Z is (CR2aR2b)r;

U is selected from the group consisting of CR5aR5b and NR6;

m is 1 or 2;

r is 0-3;

R1 is selected from the group consisting of hydrogen, optionally substituted alkyl, aralkyl, optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heteroaryl and optionally substituted heterocyclo;

each R2a, R2b, R5a and R5b are independently selected from the group consisting of hydrogen, optionally substituted alkyl, aralkyl, optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heteroaryl and optionally substituted heterocyclo;

R6 is selected from the group consisting of hydrogen, optionally substituted alkyl, aralkyl, optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heteroaryl and optionally substituted heterocyclo and COR9;

R9 is selected from the group consisting of hydrogen, optionally substituted alkyl, aralkyl, optionally substituted cycloalkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted aryl, optionally substituted heteroaryl and optionally substituted heterocyclo;

R11a, R11b, R11c and R11d are independently selected from the group consisting of hydrogen, optionally substituted alkyl, haloalkyl, aralkyl, optionally substituted cycloalkyl, optionally substituted cycloalkenyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocyclo, halo, nitro, cyano, hydroxy, amino, alkoxy, aryloxy, arylalkyloxy, alkylthio, carboxamido and sulfonamido, and

R12 is selected from the group consisting of hydrogen, optionally substituted alkyl, aralkyl, optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heteroaryl and optionally substituted heterocyclo.

The present also provides a process for preparing a compound of Formula XXI




comprising:

condensing a compound of Formula XXIII





with a compound of Formula XXIV





wherein L1 is a leaving group, to give a compound of Formula XXI,

wherein:

A1 and A2 are independently selected from the group consisting of hydrogen and optionally substituted alkyl, wherein A2 is absent when V is O;

V is selected from the group consisting of N, CH and O;

W is selected from the group consisting of CH and N;

X is selected from the group consisting of hydrogen, optionally substituted alkyl and aralkyl;

Z is (CR2aR2b)r;

U is selected from the group consisting of CR5aR5b and NR6;

m is 1 or 2;

r is 0-3;

each R2a, R2b, R5a and R5b are independently selected from the group consisting of hydrogen, optionally substituted alkyl, aralkyl, optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heteroaryl and optionally substituted heterocyclo;

R6 is selected from the group consisting of hydrogen, optionally substituted alkyl, aralkyl, optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heteroaryl and optionally substituted heterocyclo and COR9;

R9 is selected from the group consisting of hydrogen, optionally substituted alkyl, aralkyl, optionally substituted cycloalkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted aryl, optionally substituted heteroaryl and optionally substituted heterocyclo; and

T is optionally substituted heteroaryl.




BRIEF DESCRIPTION OF DRAWINGS
FIG. 1 is a graph illustrating competitive binding curves of Smac mimetics to the XIAP BIR3 domain.
FIG. 2 is a graph illustrating competitive binding curves of Smac mimetics to the cIAP1 domain.
FIG. 3 is a bar graph showing the induction of cell death by SM-1238 in the human breast cancer MDA-MB-231 and human ovarian cancer SK-OV-3 cell lines.



DETAILED DESCRIPTION OF THE INVENTION
The present invention relates to conformationally constrained compounds represented by Formulae I-XIa, which are mimetics of Smac and function as inhibitors of IAPs. Smac mimetics of Formula I-XIa display potent in vitro inhibitory activity in cancer cell lines. The invention relates to methods of sensitizing cells to inducers of apoptosis and to methods of inducing apoptosis in cells, comprising contacting the cells with a compound of Formulae I-XIa alone or in combination with an inducer of apoptosis. The invention further relates to methods of treating, ameliorating, or preventing disorders in an animal that are responsive to induction of apoptosis comprising administering to the animal a compound of Formulae I-XIa and an inducer of apoptosis. Such disorders include those characterized by a dysregulation of apoptosis and those characterized by overexpression of IAPs. The invention further relates methods of preventing or inhibiting angiogenesis in an animal in need thereof comprising administering to an animal a compound of Formulae I-XIa.
The term “IAP proteins,” as used herein, refers to any known member of the Inhibitor of Apoptosis Protein family, including, but not limited to, XIAP, cIAP-1, cIAP-2, ML-IAP, HIAP, TSIAP, KIAP, NAIP, survivin, livin, ILP-2, apollon, and BRUCE.
The term “overexpression of IAPs,” as used herein, refers to an elevated level (e.g., aberrant level) of mRNAs encoding for an IAP protein(s), and/or to elevated levels of IAP protein(s) in cells as compared to similar corresponding non-pathological cells expressing basal levels of mRNAs encoding IAP proteins or having basal levels of IAP proteins. Methods for detecting the levels of mRNAs encoding IAP proteins or levels of IAP proteins in a cell include, but are not limited to, Western blotting using IAP protein antibodies, immunohistochemical methods, and methods of nucleic acid amplification or direct RNA detection. As important as the absolute level of IAP proteins in cells is to determining that they overexpress IAP proteins, so also is the relative level of IAP proteins to other pro-apoptotic signaling molecules (e.g., pro-apoptotic Bcl-2 family proteins) within such cells. When the balance of these two are such that, were it not for the levels of the IAP proteins, the pro-apoptotic signaling molecules would be sufficient to cause the cells to execute the apoptosis program and die, said cells would be dependent on the IAP proteins for their survival. In such cells, exposure to an inhibiting effective amount of an IAP protein inhibitor will be sufficient to cause the cells to execute the apoptosis program and die. Thus, the term “overexpression of an IAP protein” also refers to cells that, due to the relative levels of pro-apoptotic signals and anti-apoptotic signals, undergo apoptosis in response to inhibiting effective amounts of compounds that inhibit the function of IAP proteins.
The terms “anticancer agent” and “anticancer drug,” as used herein, refer to any therapeutic agents (e.g., chemotherapeutic compounds and/or molecular therapeutic compounds), radiation therapies, or surgical interventions, used in the treatment of hyperproliferative diseases such as cancer (e.g., in mammals).
The term “prodrug,” as used herein, refers to a pharmacologically inactive derivative of a parent “drug” molecule that requires biotransformation (e.g., either spontaneous or enzymatic) within the target physiological system to release, or to convert (e.g., enzymatically, physiologically, mechanically, electromagnetically) the prodrug into the active drug. Prodrugs are designed to overcome problems associated with stability, toxicity, lack of specificity, or limited bioavailability. Exemplary prodrugs comprise an active drug molecule itself and a chemical masking group (e.g., a group that reversibly suppresses the activity of the drug). Some preferred prodrugs are variations or derivatives of compounds that have groups cleavable under metabolic conditions. Exemplary prodrugs become pharmaceutically active in vivo or in vitro when they undergo solvolysis under physiological conditions or undergo enzymatic degradation or other biochemical transformation (e.g., phosphorylation, hydrogenation, dehydrogenation, glycosylation). Prodrugs often offer advantages of solubility, tissue compatibility, or delayed release in the mammalian organism. (See e.g., Bundgard, Design of Prodrugs, pp. 7-9, 21-24, Elsevier, Amsterdam (1985); and Silverman, The Organic Chemistry of Drug Design and Drug Action, pp. 352-401, Academic Press, San Diego, Calif. (1992)). Common prodrugs include acid derivatives such as esters prepared by reaction of parent acids with a suitable alcohol (e.g., a lower alkanol), amides prepared by reaction of the parent acid compound with an amine, or basic groups reacted to form an acylated base derivative (e.g., a lower alkylamide).
The term “pharmaceutically acceptable salt,” as used herein, refers to any salt (e.g., obtained by reaction with an acid or a base) of a compound of the present invention that is physiologically tolerated in the target animal (e.g., a mammal). Salts of the compounds of the present invention may be derived from inorganic or organic acids and bases. Examples of acids include, but are not limited to, hydrochloric, hydrobromic, sulfuric, nitric, perchloric, fumaric, maleic, phosphoric, glycolic, lactic, salicylic, succinic, toluene-p-sulfonic, tartaric, acetic, citric, methanesulfonic, ethanesulfonic, formic, benzoic, malonic, sulfonic, naphthalene-2-sulfonic, benzenesulfonic acid, and the like. Other acids, such as oxalic, while not in themselves pharmaceutically acceptable, may be employed in the preparation of salts useful as intermediates in obtaining the compounds of the invention and their pharmaceutically acceptable acid addition salts.
Examples of bases include, but are not limited to, alkali metal (e.g., sodium) hydroxides, alkaline earth metal (e.g., magnesium) hydroxides, ammonia, and compounds of formula NW4+, wherein W is C1-4 alkyl, and the like.
Examples of salts include, but are not limited to: acetate, adipate, alginate, aspartate, benzoate, benzenesulfonate, bisulfate, butyrate, citrate, camphorate, camphorsulfonate, cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate, fumarate, flucoheptanoate, glycerophosphate, hemisulfate, heptanoate, hexanoate, chloride, bromide, iodide, 2-hydroxyethanesulfonate, lactate, maleate, mesylate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, oxalate, palmoate, pectinate, persulfate, phenylpropionate, picrate, pivalate, propionate, succinate, tartrate, thiocyanate, tosylate, undecanoate, and the like. Other examples of salts include anions of the compounds of the present invention compounded with a suitable cation such as Na+, NH4+, and NW4+ (wherein W is a C1-4 alkyl group), and the like. For therapeutic use, salts of the compounds of the present invention are contemplated as being pharmaceutically acceptable. However, salts of acids and bases that are non-pharmaceutically acceptable may also find use, for example, in the preparation or purification of a pharmaceutically acceptable compound.
The term “therapeutically effective amount,” as used herein, refers to that amount of the therapeutic agent sufficient to result in amelioration of one or more symptoms of a disorder, or prevent advancement of a disorder, or cause regression of the disorder. For example, with respect to the treatment of cancer, a therapeutically effective amount preferably refers to the amount of a therapeutic agent that decreases the rate of tumor growth, decreases tumor mass, decreases the number of metastases, increases time to tumor progression, or increases survival time by at least 5%, preferably at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or at least 100%.
The terms “sensitize” and “sensitizing,” as used herein, refer to making, through the administration of a first agent (e.g., a compound of Formula I), an animal or a cell within an animal more susceptible, or more responsive, to the biological effects (e.g., promotion or retardation of an aspect of cellular function including, but not limited to, cell division, cell growth, proliferation, invasion, angiogenesis, or apoptosis) of a second agent. The sensitizing effect of a first agent on a target cell can be measured as the difference in the intended biological effect (e.g., promotion or retardation of an aspect of cellular function including, but not limited to, cell growth, proliferation, invasion, angiogenesis, or apoptosis) observed upon the administration of a second agent with and without administration of the first agent. The response of the sensitized cell can be increased by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 100%, at least 150%, at least 200%, at least 350%, at least 300%, at least 350%, at least 400%, at least 450%, or at least 500% over the response in the absence of the first agent.
The term “dysregulation of apoptosis,” as used herein, refers to any aberration in the ability of (e.g., predisposition) a cell to undergo cell death via apoptosis. Dysregulation of apoptosis is associated with or induced by a variety of conditions, including for example, autoimmune disorders (e.g., systemic lupus erythematosus, rheumatoid arthritis, graft-versus-host disease, myasthenia gravis, or Sjögren's syndrome), chronic inflammatory conditions (e.g., psoriasis, asthma or Crohn's disease), hyperproliferative disorders (e.g., tumors, B cell lymphomas, or T cell lymphomas), viral infections (e.g., herpes, papilloma, or HIV), and other conditions such as osteoarthritis and atherosclerosis. It should be noted that when the dysregulation is induced by or associated with a viral infection, the viral infection may or may not be detectable at the time dysregulation occurs or is observed. That is, viral-induced dysregulation can occur even after the disappearance of symptoms of viral infection.
The term “angiogenesis,” as used herein means the generation of new blood vessels into a tissue or organ. The term “antiangiogenesis,” as used herein, refers to prevention or reduction of the growth of new blood vessels. Examples of diseases or disorders associated with angiogenesis that may be treated with the compounds of the invention include macular degeneration, rheumatoid arthritis, psoriasis, diabetic retinopathy, retinopathy of prematurity, corneal graft rejection, neovascular glaucoma, retrolental fibroplasia, rubeosis, Osler-Webber Syndrome, myocardial angiogenesis, plaque neovascularization, telangiectasia, hemophiliac joints, angiofibroma, wound granulation, intestinal adhesions, atherosclerosis, scleroderma and hypertrophic scars.
The term “hyperproliferative disease,” as used herein, refers to any condition in which a localized population of proliferating cells in an animal is not governed by the usual limitations of normal growth. Examples of hyperproliferative disorders include, but are not restricted to cancers (e.g., tumors, neoplasms, lymphomas and the like) or autoimmune disorders. A neoplasm is said to be benign if it does not undergo invasion or metastasis and malignant if it does either of these. A “metastatic” cell means that the cell can invade and destroy neighboring body structures. Hyperplasia is a form of cell proliferation involving an increase in cell number in a tissue or organ without significant alteration in structure or function. Metaplasia is a form of controlled cell growth in which one type of fully differentiated cell substitutes for another type of differentiated cell. In another embodiment, the hyperproliferative disease is rheumatoid arthritis, inflammatory bowel disease, osteoarthritis, leiomyomas, adenomas, lipomas, hemangiomas, fibromas, vascular occlusion, restenosis, atherosclerosis, pre-neoplastic lesions (such as adenomatous hyperplasia and prostatic intraepithelial neoplasia), carcinoma in situ, oral hairy leukoplakia, or psoriasis.
The pathological growth of activated lymphoid cells often results in an autoimmune disorder or a chronic inflammatory condition. As used herein, the term “autoimmune disorder” refers to any condition in which an organism produces antibodies or immune cells which recognize the organism's own molecules, cells or tissues. Non-limiting examples of autoimmune disorders include autoimmune hemolytic anemia, autoimmune hepatitis, Berger's disease or IgA nephropathy, celiac sprue, chronic fatigue syndrome, Crohn's disease, dermatomyositis, fibromyalgia, graft versus host disease, Grave's disease, Hashimoto's thyroiditis, idiopathic thrombocytopenia purpura, lichen planus, multiple sclerosis, myasthenia gravis, psoriasis, rheumatic fever, rheumatic arthritis, scleroderma, Sjögren's syndrome, systemic lupus erythematosus, type 1 diabetes, ulcerative colitis, vitiligo, and the like.
The term “neoplastic disease,” as used herein, refers to any abnormal growth of cells being either benign (non-cancerous) or malignant (cancerous).
The term “anti-neoplastic agent,” as used herein, refers to any compound that retards the proliferation, growth, or spread of a targeted (e.g., malignant) neoplasm.
The terms “prevent,” “preventing,” and “prevention,” as used herein, refer to a decrease in the occurrence of pathological cells (e.g., hyperproliferative or neoplastic cells) in an animal. The prevention may be complete, e.g., the total absence of pathological cells in a subject. The prevention may also be partial, such that the occurrence of pathological cells in a subject is less than that which would have occurred without the present invention.
The term “apoptosis-modulating agents,” as used herein, refers to agents which are involved in modulating (e.g., inhibiting, decreasing, increasing, promoting) apoptosis. In one embodiment, the apoptosis-modulating agent is an inducer of apoptosis. The term “inducer of apoptosis,” as used herein, refers to an agent that induces apoptosis in cells (e.g., cancer cells), rendering those cells more susceptible to executing the apoptosis program. In one embodiment, an agent that induces apoptosis is an anticancer agent. Examples of apoptosis-modulating agents include proteins which comprise a death domain such as, but not limited to, Fas/CD95, TRAMP, TNF R1, DR1, DR2, DR3, DR4, DR5, DR6, FADD, and RIP. Other examples of apoptotic-modulating agents include, but are not limited to, TNFα, Fas ligand, antibodies to Fas/CD95 and other TNF family receptors, TRAIL (also known as Apo2 Ligand or Apo2L/TRAIL), agonists (e.g., monoclonal or polyclonal agonistic antibodies) of TRAIL-R1 or TRAIL-R2, Bcl-2, p53, BAX, BAD, Akt, CAD, PI3 kinase, PP1, and caspase proteins. Modulating agents broadly include agonists and antagonists of TNF family receptors and TNF family ligands. Apoptosis-modulating agents may be soluble or membrane bound (e.g. ligand or receptor). Preferred apoptosis-modulating agents are inducers of apoptosis, such as TNF or a TNF-related ligand, particularly a TRAMP ligand, a Fas/CD95 ligand, a TNFR-1 ligand, or TRAIL
The inhibitors of IAPs of the present invention are Smac mimetics having the general Formula I:




wherein:

A1 and A2 are independently selected from the group consisting of hydrogen and optionally substituted alkyl, wherein A2 is absent when V is O;

V is selected from the group consisting of N, CH and O;

W is selected from the group consisting of CH and N;

X is selected from the group consisting of hydrogen, optionally substituted alkyl and aralkyl;

Y is selected from the group consisting of CON(R1), N(R1)CO, C(O)O, OC(O), (CH2)1-3, wherein one or more CH2 groups can be replaced by O, S, or NR1, optionally substituted aryl and optionally substituted heteroaryl;

Z is (CR2aR2b)r;

D is (CR3aR3b)n—U—(cR4aR4b)m;

U is selected from the group consisting of CR5aR5b and NR6;

J is (CR7aR7b)p-L-(CR8aR8b)q;

T is optionally substituted heteroaryl;

n, m, p and q are independently selected from the group consisting of 0-5;

r is 0-3;

R1 is selected from the group consisting of hydrogen, optionally substituted alkyl, aralkyl, optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heteroaryl and optionally substituted heterocyclo;

each R2a, R2b, R3a, R3b, R4a, R4b, R5a, R5b, R7a, R7b, R8a and R8b are independently selected from the group consisting of hydrogen, optionally substituted alkyl, aralkyl, optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heteroaryl and optionally substituted heterocyclo;

R6 is selected from the group consisting of hydrogen, optionally substituted alkyl, aralkyl, optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heteroaryl and optionally substituted heterocyclo and COR9;

L is selected from the group consisting of O, S, NR1, NCOR9, CR7aR7b, C═O, C═S and C═NR1; and

R9 is selected from the group consisting of hydrogen, optionally substituted alkyl, aralkyl, optionally substituted cycloalkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted aryl, optionally substituted heteroaryl and optionally substituted heterocyclo;

or pharmaceutically acceptable salt or prodrug thereof.

In another embodiment, Smac mimetics are compounds of Formula I wherein n is 1, m is 1 or 2, p is 0, L is CH2 and q is 1. In another embodiment, Smac mimetics are compounds of Formula I wherein R3a, R3b, R4a, R4b, R7a, R7b, R8a and R8b are hydrogen. In another embodiment, Smac mimetics are compounds of Formula I wherein Y is CON(H), W is CH, r is 0 and V is N. In another embodiment, Smac mimetics are compounds of Formula I wherein Y is CON(H), W is CH, r is 1, V is N, R2a is optionally substituted alkyl, R2b is hydrogen and X is hydrogen.
In another particular embodiment, Smac mimetics are compounds of Formula II:




wherein A1, A2, V, Z, W, X, Y, D, J and T having the meanings as described above for Formula I; or a pharmaceutically acceptable salt or prodrug thereof.

In one embodiment, Smac mimetics are compounds of Formula II wherein n is 1, m is 1 or 2, p is 0, L is CH2 and q is 1. In another embodiment, Smac mimetics are compounds of Formula II wherein R3a, R3b, R4a, R4b, R7a, R7b, R8a and R8b are hydrogen. In another embodiment, Smac mimetics are compounds of Formula II wherein Y is CON(H), W is CH, r is 0 and V is N. In another embodiment, Smac mimetics are compounds of Formula II wherein Y is CON(H), W is CH, r is 1, V is N, R2a is optionally substituted alkyl, R2b is hydrogen and X is hydrogen.
In another particular embodiment, Smac mimetics are compounds of Formula III:




wherein A1, A2, V, Z, W, X, Y, U, and T have the meanings as described above for Formula I, and m is 1 or 2; or a pharmaceutically acceptable salt or prodrug thereof.

In another embodiment, Smac mimetics are compounds of Formula III wherein Y is CON(H), W is CH, r is 1, V is N, R2a is optionally substituted alkyl, R2b is hydrogen and X is hydrogen. In one embodiment, m is 1. In another embodiment, m is 2.
In another particular embodiment, Smac mimetics are compounds of Formula IV:




wherein A1, A2, X, U, and T have the meanings as described above for Formula I, and m is 1 or 2; or a pharmaceutically acceptable salt or prodrug thereof.

In another embodiment, Smac mimetics are compounds of Formula IV wherein m is 1. In another embodiment, m is 2.
In another particular embodiment, Smac mimetics are compounds of Formula V:




wherein A1, A2, X, U and T have the meanings as described above for Formula I, and m is 1 or 2; or a pharmaceutically acceptable salt or prodrug thereof.

In another embodiment, Smac mimetics are compounds of Formula V wherein A1 is optionally substituted alkyl and A2 is hydrogen. In another embodiment, X is optionally substituted alkyl. In another embodiment, U is CH2. In another embodiment, U is NR6. In another embodiment, R6 is COR9. In another embodiment, R9 is optionally substituted alkyl or aralkyl. In another embodiment, R9 is —CH2CH(CH3)2. In another embodiment, m is 2. In another embodiment, m is 1.
In another particular embodiment, Smac mimetics are compounds of Formula VI:




wherein A1 and X are optionally substituted alkyl and T is optionally substituted heteroaryl; or a pharmaceutically acceptable salt or prodrug thereof.

In one embodiment, A1 and X are independently optionally substituted C1-C4 alkyl. In another embodiment, A1 and X are independently C1-C4 alkyl. In another embodiment, A1 and X are independently selected from the group consisting of methyl and ethyl. In one embodiment, A1 and X are methyl. In one embodiment, A1 is hydroxyalkyl and X is optionally substituted alkyl. In another embodiment, A1 is HOCH2CH2—.
In another particular embodiment, Smac mimetics are compounds of Formula VII:




wherein A1 and X are optionally substituted alkyl, R9 is optionally substituted alkyl or aralkyl, and T is optionally substituted heteroaryl; or a pharmaceutically acceptable salt or pro drug thereof.

In one embodiment, A1 and X are independently optionally substituted C1-C4 alkyl. In another embodiment, A1 and X are independently C1-C4 alkyl. In another embodiment, A1 and X are independently selected from the group consisting of methyl and ethyl. In one embodiment, A1 and X are methyl. In another embodiment, A1 is hydroxyalkyl and X is optionally substituted alkyl. In another embodiment, A1 is HOCH2CH2—.
In another particular embodiment, Smac mimetics are compounds of Formulae I-VII wherein T is




wherein Q is O, S or NR12, R12 is hydrogen, optionally substituted alkyl, aralkyl, optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heteroaryl or optionally substituted heterocyclo, R10a, R10b, R11a, R11b, R11c and R11d are independently selected from the group consisting of hydrogen, optionally substituted alkyl, haloalkyl, aralkyl, optionally substituted cycloalkyl, optionally substituted cycloalkenyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocyclo, halo, nitro, cyano, hydroxy, amino, alkoxy, aryloxy, arylalkyloxy, alkylthio, carboxamido and sulfonamido, Z1, Z2, and Z3 are independently CR11e or N, wherein at least one of Z1, Z2, and Z3 is CR11e and at least one of Z1, Z2, and Z3 is N, and R11e is selected from the group consisting of hydrogen, optionally substituted alkyl, haloalkyl, aralkyl, optionally substituted cycloalkyl, optionally substituted cycloalkenyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocyclo, halo, nitro, cyano, hydroxy, amino, alkoxy, aryloxy, arylalkyloxy, alkylthio, carboxamido and sulfonamido; or a pharmaceutically acceptable salt or prodrug thereof.

In one embodiment, at least one of R10a, R10b, R11a, R11b, R11c and R11d is optionally substituted phenyl, aralkyl, or optionally substituted alkyl. In one embodiment, R12 is hydrogen or optionally substituted alkyl. In one embodiment, R12 is hydrogen. In one embodiment, R10a is optionally substituted phenyl. In one embodiment, R11a is optionally substituted phenyl.
In another particular embodiment, Smac mimetics are compounds of Formulae I-VII wherein T is




wherein Q, Z1, Z2, Z3, R10a, R10b, R11a, R11b, R11c and R11d have the meanings described above; or a pharmaceutically acceptable salt or prodrug thereof.

In another particular embodiment, Smac mimetics are compounds of Formulae I-VII wherein T is




wherein Q, R10a and R10b have the meanings described above; or a pharmaceutically acceptable salt or prodrug thereof.

In one embodiment, R10a is optionally substituted aryl, aralkyl, or optionally substituted alkyl. In one embodiment, R10b is hydrogen. In another embodiment, R10a is optionally substituted aryl and R10b is hydrogen. In one embodiment, Q is S. In one embodiment, Q is O. In one embodiment, Q is NR12. In one embodiment, R12 is hydrogen or optionally substituted alkyl. In another embodiment R12 is hydrogen.
In another particular embodiment, Smac mimetics are compounds of Formulae I-VII wherein T is




wherein Q, R11a, R11b, R11c and R11d have the meanings described above; or a pharmaceutically acceptable salt or prodrug thereof.

In one embodiment, R11a is optionally substituted aryl, aralkyl, or optionally substituted alkyl. In one embodiment, R11b, R11c and R11d are hydrogen. In another embodiment, R11a is optionally substituted aryl and R11b, R11c and R11d are hydrogen. In another embodiment, R11a is optionally substituted aryl and R11a, R11b, and R11c are hydrogen. In one embodiment, Q is S. In one embodiment, Q is O. In one embodiment, Q is NR12. In one embodiment, R12 is hydrogen or optionally substituted alkyl. In another embodiment, R12 is hydrogen.
In another particular embodiment, Smac mimetics are compounds of Formulae I-VII wherein T is




wherein Q, Z1, Z2, Z3, and R11a have the meanings described above; or a pharmaceutically acceptable salt or prodrug thereof.

In one embodiment, R11a is optionally substituted aryl, aralkyl, or optionally substituted alkyl and at least one of Z1, Z2, and Z3 is N. In one embodiment, R11a is optionally substituted aryl. In one embodiment, Z1 is N and Z2 and Z3 are CH. In one embodiment, Z2 is N and Z1 and Z3 are CH. In one embodiment, Z3 is N and Z1 and Z2 are CH. In one embodiment, Z1 and Z3 are N and Z2 is CH. In one embodiment, Q is S. In one embodiment, Q is O. In one embodiment, Q is NR12. In one embodiment, R12 is hydrogen or optionally substituted alkyl. In another embodiment, R12 is hydrogen.
In another particular embodiment, Smac mimetics are compounds of Formula VIII:




wherein A1 and X are optionally substituted alkyl, m is 1 or 2, Q is O, S or NR12, R12 is hydrogen, optionally substituted alkyl, aralkyl, optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heteroaryl or optionally substituted heterocyclo, and R10a and R10b are independently selected from the group consisting of hydrogen, optionally substituted alkyl, haloalkyl, aralkyl, optionally substituted cycloalkyl, optionally substituted cycloalkenyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocyclo, halo, nitro, cyano, hydroxy, amino, alkoxy, aryloxy, arylalkyloxy, alkylthio, carboxamido and sulfonamido; or a pharmaceutically acceptable salt or prodrug thereof.

In one embodiment, m is 2. In another embodiment, m is 1. In one embodiment, R10a is optionally substituted aryl. In another embodiment, R10b is hydrogen. In another embodiment, R10a is optionally substituted aryl and R10b is hydrogen. In one embodiment, R9 is hydrogen or optionally substituted alkyl. In one embodiment, Q is S.
In another particular embodiment, Smac mimetics are compounds of Formula IX:




wherein A1 and X are optionally substituted alkyl, m is 1 or 2, Q is O, S or NR12, R12 is hydrogen, optionally substituted alkyl, aralkyl, optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heteroaryl or optionally substituted heterocyclo, and R11a, R11b, R11c and R11d are independently selected from the group consisting of hydrogen, optionally substituted alkyl, haloalkyl, aralkyl, optionally substituted cycloalkyl, optionally substituted cycloalkenyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocyclo, halo, nitro, cyano, hydroxy, amino, alkoxy, aryloxy, arylalkyloxy, alkylthio, carboxamido and sulfonamido; or a pharmaceutically acceptable salt or prodrug thereof.

In one embodiment, m is 2. In another embodiment, m is 1. In one embodiment, R11a is optionally substituted aryl, aralkyl, or optionally substituted alkyl. In one embodiment, R11b, R11c and R11d are hydrogen. In another embodiment, R11a is optionally substituted aryl and R11b, R11c and R11d are hydrogen. In another embodiment, R11d is optionally substituted aryl and R11a, R11b and R11c are hydrogen. In one embodiment, Q is NR12. In one embodiment, R12 is hydrogen or optionally substituted alkyl. In another embodiment, R12 is hydrogen. In one embodiment, Q is S. In one embodiment, Q is O.
In another particular embodiment, Smac mimetics are compounds of Formula X:




wherein A1 and X are optionally substituted alkyl, R9 is optionally substituted alkyl or aralkyl, m is 1 or 2, Q is O, S or NR12, R12 is hydrogen, optionally substituted alkyl, aralkyl, optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heteroaryl or optionally substituted heterocyclo, and R10a and R10b are independently selected from the group consisting of hydrogen, optionally substituted alkyl, haloalkyl, aralkyl, optionally substituted cycloalkyl, optionally substituted cycloalkenyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocyclo, halo, nitro, cyano, hydroxy, amino, alkoxy, aryloxy, arylalkyloxy, alkylthio, carboxamido and sulfonamido; or a pharmaceutically acceptable salt or prodrug thereof.

In one embodiment, m is 2. In another embodiment, m is 1. In one embodiment, R10a is optionally substituted aryl. In one embodiment, R10b is hydrogen. In another embodiment, R10a is optionally substituted aryl and R10b is hydrogen. In one embodiment, R12 is hydrogen or optionally substituted alkyl. In one embodiment, R9 is —CH2CH(CH3)2. In one embodiment, Q is S.
In another particular embodiment, Smac mimetics are compounds of Formula XI:




wherein A1 and X are optionally substituted alkyl, R9 is optionally substituted alkyl or aralkyl, m is 1 or 2, Q is O, S or NR12, R12 is hydrogen, optionally substituted alkyl, aralkyl, optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heteroaryl or optionally substituted heterocyclo, and R11a, R11b, R11c and R11d are independently selected from the group consisting of hydrogen, optionally substituted alkyl, haloalkyl, aralkyl, optionally substituted cycloalkyl, optionally substituted cycloalkenyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocyclo, halo, nitro, cyano, hydroxy, amino, alkoxy, aryloxy, arylalkyloxy, alkylthio, carboxamido and sulfonamido; or a pharmaceutically acceptable salt or prodrug thereof.

In one embodiment, m is 2. In another embodiment, m is 1. In one embodiment, R11a is optionally substituted aryl, aralkyl, or optionally substituted alkyl. In one embodiment, R11b, R11c and R11d are hydrogen. In another embodiment, R11a is optionally substituted aryl and R11b, R11b and R11c are hydrogen. In another embodiment, R11d is optionally substituted aryl and R11a, R11b, and R11c are hydrogen. In one embodiment, Q is NR12. In one embodiment, R12 is hydrogen or optionally substituted alkyl. In another embodiment, R12 is hydrogen. In one embodiment, Q is S. In one embodiment, Q is O. In one embodiment, R9 is —CH2CH(CH3)2.
In another particular embodiment, Smac mimetics are compounds of Formula X wherein A1 and X are methyl, m is 1, R9 is optionally substituted alkyl or aralkyl, R10a is optionally substituted aryl, R10b is hydrogen, and Q is S.
In another particular embodiment, Smac mimetics are compounds of Formula XIa:




wherein R9 is optionally substituted alkyl or aralkyl and R11a is optionally substituted aryl.

The term “alkyl” as used herein by itself or part of another group refers to a straight-chain or branched saturated aliphatic hydrocarbon having from one to eighteen carbons or the number of carbons designated (e.g., C1-C18 means 1 to 18 carbons). In one embodiment, the alkyl is a C1-C8 alkyl. In another embodiment, the alkyl is a C1-C6 alkyl. In another embodiment, the alkyl is a C1-C4 alkyl. Exemplary alkyl groups include methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl, n-pentyl, n-hexyl, isohexyl, n-heptyl, 4,4-dimethylpentyl, n-octyl, 2,2,4-trimethylpentyl, nonyl, decyl and the like.
The term “optionally substituted alkyl” as used herein by itself or part of another group means that the alkyl as defined above is either unsubstituted or substituted with one, two or three substituents independently selected from hydroxy (i.e., —OH), nitro (i.e., —NO2), cyano (i.e., —CN), optionally substituted cycloalkyl, optionally substituted heteroaryl, optionally substituted heterocyclo, alkoxy, aryloxy, aralkyloxy, alkylthio, carboxamido or sulfonamido. In one embodiment, the optionally substituted alkyl is substituted with two substituents. In another embodiment, the optionally substituted alkyl is substituted with one substituents. In another embodiment, the substituents are selected from hydroxyl (i.e., a hydroxyalkyl) or amino (i.e., an aminoalkyl). Exemplary optionally substituted alkyl groups include —CH2OCH3, —CH2CH2NH2, —CH2CH2CN, —CH2SO2CH3, hydroxymethyl, hydroxyethyl, hydroxypropyl and the like.
The term “alkylenyl” as used herein by itself or part of another group refers to a divalent alkyl radical containing one, two, three or four joined methylene groups. Exemplary alkylenyl groups include —(CH2)—, —(CH2)2—, —(CH2)3— and —(CH2)4—.
The term “haloalkyl” as used herein by itself or part of another group refers to an alkyl as defined above having one to six halo substituents. In one embodiment, the haloalkyl has one, two or three halo substituents. Exemplary haloalkyl groups include trifluoromethyl, —CH2CH2F and the like.
The term “hydroxyalkyl” as used herein by itself or part of another group refers to an alkyl as defined above having one, two or three hydroxy substituents. In one embodiment, the hydroxyalkyl has one hydroxy substituent. Exemplary hydroxyalkyl groups include hydroxymethyl, hydroxyethyl, hydroxypropyl and the like.
The term “aralkyl” as used herein by itself or part of another group refers to an optionally substituted alkyl as defined above having one, two or three optionally substituted aryl substituents. In one embodiment, the aralkyl has two optionally substituted aryl substituents. In another embodiment, the aralkyl has one optionally substituted aryl substituent. In another embodiment, the aralkyl is an aryl(C1-C4 alkyl). In another embodiment, the aryl(C1-C4 alkyl) has two optionally substituted aryl substituents. In another embodiment, the aryl(C1-C4 alkyl) has one optionally substituted aryl substituent. Exemplary aralkyl groups include, for example, benzyl, phenylethyl, (4-fluorophenyl)ethyl, phenylpropyl, diphenylmethyl (i.e., Ph2CH—), diphenylethyl (Ph2CHCH2—) and the like.
The term “cycloalkyl” as used herein by itself or part of another group refers to saturated and partially unsaturated (containing one or two double bonds) cyclic hydrocarbon groups containing one to three rings having from three to twelve carbon atoms (i.e., C3-C12 cycloalkyl) or the number of carbons designated. In one embodiment, the cycloalkyl has one ring. In another embodiment, the cycloalkyl is a C3-C7 cycloalkyl. Exemplary cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, norbornyl, decalin, adamantyl and the like.
The term “optionally substituted cycloalkyl” as used herein by itself or part of another group means the cycloalkyl as defined above is either unsubstituted or substituted with one, two or three substituents independently selected from halo, nitro, cyano, hydroxy, amino, optionally substituted alkyl, haloalkyl, hydroxyalkyl, aralkyl, optionally substituted cycloalkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocyclo, alkoxy, aryloxy, aralkyloxy, alkylthio, carboxamido or sulfonamido. The term “optionally substituted cycloalkyl” also means the cycloalkyl as defined above may be fused to an optionally substituted aryl. Exemplary optionally substituted cycloalkyl groups include




and the like.

The term “alkenyl” as used herein by itself or part of another group refers to an alkyl group as defined above containing one, two or three carbon-to-carbon double bonds. In one embodiment, the alkenyl has one carbon-to-carbon double bond. Exemplary alkenyl groups include —CH═CH2, —CH2CH═CH2, —CH2CH2CH═CH2, —CH2CH2CH═CHCH3 and the like.
The term “optionally substituted alkenyl” as used herein by itself or part of another group means the alkenyl as defined above is either unsubstituted or substituted with one, two or three substituents independently selected from halo, nitro, cyano, hydroxy, amino, optionally substituted alkyl, haloalkyl, hydroxyalkyl, aralkyl, optionally substituted cycloalkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocyclo, alkoxy, aryloxy, aralkyloxy, alkylthio, carboxamido or sulfonamido. Exemplary optionally substituted alkenyl groups include —CH═CHPh, —CH2CH═CHPh and the like.
The term “cycloalkenyl” as used herein by itself or part of another group refers to a cycloalkyl group as defined above containing one, two or three carbon-to-carbon double bonds. In one embodiment, the cycloalkenyl has one carbon-to-carbon double bond. Exemplary cycloalkenyl groups include cyclopentene, cyclohexene and the like.
The term “optionally substituted cycloalkenyl” as used herein by itself or part of another group means the cycloalkenyl as defined above is either unsubstituted or substituted with one, two or three substituents independently selected from halo, nitro, cyano, hydroxy, amino, optionally substituted alkyl, haloalkyl, hydroxyalkyl, aralkyl, optionally substituted cycloalkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocyclo, alkoxy, aryloxy, aralkyloxy, alkylthio, carboxamido or sulfonamido.
The term “alkynyl” as used herein by itself or part of another group refers to an alkyl group as defined above containing one to three carbon-to-carbon triple bonds. In one embodiment, the alkynyl has one carbon-to-carbon triple bond. Exemplary alkynyl groups include —C≡CH, —C≡CCH3, —CH2C≡CH, —CH2CH2C≡CH and —CH2CH2C≡CCH3.
The term “optionally substituted alkynyl” as used herein by itself or part of another group means the alkynyl as defined above is either unsubstituted or substituted with one, two or three substituents independently selected from halo, nitro, cyano, hydroxy, amino, optionally substituted alkyl, haloalkyl, hydroxyalkyl, aralkyl, optionally substituted cycloalkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocyclo, alkoxy, aryloxy, aralkyloxy, alkylthio, carboxamido or sulfonamido. Exemplary optionally substituted alkenyl groups include —C≡CPh, —CH2C≡CPh and the like.
The term “aryl” as used herein by itself or part of another group refers to monocyclic and bicyclic aromatic ring systems having from six to fourteen carbon atoms (i.e., C6-C14 aryl) such as phenyl (abbreviated as Ph), 1-naphthyl and 2-naphthyl and the like.
The term “optionally substituted aryl” as used herein by itself or part of another group means the aryl as defined above is either unsubstituted or substituted with one to five substituents independently selected from halo, nitro, cyano, hydroxy, amino, optionally substituted alkyl, haloalkyl, hydroxyalkyl, aralkyl, optionally substituted cycloalkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocyclo, alkoxy, aryloxy, aralkyloxy, alkylthio, carboxamido or sulfonamido. In one embodiment, the optionally substituted aryl is an optionally substituted phenyl. In one embodiment, the optionally substituted phenyl has four substituents. In another embodiment, the optionally substituted phenyl has three substituents. In another embodiment, the optionally substituted phenyl has two substituents. In another embodiment, the optionally substituted phenyl has one substituent. Exemplary substituted aryl groups include 2-methylphenyl, 2-methoxyphenyl, 2-fluorophenyl, 2-chlorophenyl, 2-bromophenyl, 3-methylphenyl, 3-methoxyphenyl, 3-fluorophenyl, 3-chlorophenyl, 4-methylphenyl, 4-ethylphenyl, 4-methoxyphenyl, 4-fluorophenyl, 4-chlorophenyl, 2,6-di-fluorophenyl, 2,6-di-chlorophenyl, 2-methyl, 3-methoxyphenyl, 2-ethyl, 3-methoxyphenyl, 3,4-di-methoxyphenyl, 3,5-di-fluorophenyl 3,5-di-methylphenyl and 3,5-dimethoxy, 4-methylphenyl and the like. The term optionally substituted aryl is meant to include groups having fused optionally substituted cycloalkyl and fused optionally substituted heterocyclo rings. Examples include




and the like.

The term “heteroaryl” as used herein by itself or part of another group refers to monocyclic and bicyclic aromatic ring systems having from five to fourteen carbon atoms (i.e., C5-C14 heteroaryl) and one, two, three or four heteroatoms independently selected from the group consisting of oxygen, nitrogen and sulfur. In one embodiment, the heteroaryl has three heteroatoms. In one embodiment, the heteroaryl has two heteroatoms. In one embodiment, the heteroaryl has one heteroatom. Exemplary heteroaryl groups include 1-pyrrolyl, 2-pyrrolyl, 3-pyrrolyl, 2-imidazolyl, 4-imidazolyl, pyrazinyl, 2-oxazolyl, 4-oxazolyl, 5-oxazolyl, 3-isoxazolyl, 4-isoxazolyl, 5-isoxazolyl, 2-thiazolyl, 4-thiazolyl, 5-thiazolyl, 2-furyl, 3-furyl, 2-thienyl, 3-thienyl, 2-pyridyl, 3-pyridyl, 4-pyridyl, 2-pyrimidyl, 4-pyrimidyl, purinyl, 2-benzimidazolyl, 4-benzimidazolyl, 5-benzimidazolyl, 2-benzthiazolyl, 4-benzthiazolyl, 5-benzthiazolyl, 5-indolyl, 3-indazolyl, 4-indazolyl, 5-indazolyl, 1-isoquinolyl, 5-isoquinolyl, 2-quinoxalinyl, 5-quinoxalinyl, 2-quinolyl 3-quinolyl, 6-quinolyl and the like. The term heteroaryl is meant to include possible N-oxides. Exemplary N-oxides include pyridyl N-oxide and the like.
The term “optionally substituted heteroaryl” as used herein by itself or part of another group means the heteroaryl as defined above is either unsubstituted or substituted with one to four substituents, typically one or two substituents, independently selected from halo, nitro, cyano, hydroxy, amino, optionally substituted alkyl, haloalkyl, hydroxyalkyl, aralkyl, optionally substituted cycloalkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocyclo, alkoxy, aryloxy, aralkyloxy, alkylthio, carboxamido or sulfonamido. In one embodiment, the optionally substituted heteroaryl has one substituent. In another embodiment, the substituent is an optionally substituted aryl, aralkyl, or optionally substituted alkyl. In another embodiment, the substituent is an optionally substituted phenyl. Any available carbon or nitrogen atom my be substituted. Exemplary optionally substituted heteroaryl groups include




and the like.

The term “heterocyclo” as used herein by itself or part of another group refers to saturated and partially unsaturated (containing one or two double bonds) cyclic groups containing one to three rings having from two to twelve carbon atoms (i.e., C2-C12 heterocyclo) and one or two oxygen, sulfur or nitrogen atoms. The heterocyclo can be optionally linked to the rest of the molecule through a carbon or nitrogen atom. Exemplary heterocyclo groups include




and the like.

The term “optionally substituted heterocyclo” as used herein by itself or part of another group means the heterocyclo as defined above is either unsubstituted or substituted with one to four substituents independently selected from halo, nitro, cyano, hydroxy, amino, optionally substituted alkyl, haloalkyl, hydroxyalkyl, aralkyl, optionally substituted cycloalkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocyclo, alkoxy, aryloxy, aralkyloxy, alkylthio, carboxamido, sulfonamido, —CORc, —SO2Rd, —N(Re)CORf, —N(Re)SO2Rg or —N(Re)C═N(Rh)-amino. Substitution may occur on any available carbon or nitrogen atom. Exemplary substituted heterocyclo groups include




and the like. An optionally substituted heterocyclo may be fused to an aryl group to provide an optionally substituted aryl as described above.

The term “alkoxy” as used herein by itself or part of another group refers to a haloalkyl, optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted alkenyl or optionally substituted alkynyl attached to a terminal oxygen atom. Exemplary alkoxy groups include methoxy, tert-butoxy, —OCH2CH═CH2 and the like.
The term “aryloxy” as used herein by itself or part of another group refers to an optionally substituted aryl attached to a terminal oxygen atom. Exemplary aryloxy groups include phenoxy and the like.
The term “aralkyloxy” as used herein by itself or part of another group refers to an aralkyl attached to a terminal oxygen atom. Exemplary aralkyloxy groups include benzyloxy and the like.
The term “alkylthio” as used herein by itself or part of another group refers to a haloalkyl, aralkyl, optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted alkenyl or optionally substituted alkynyl attached to a terminal sulfur atom. Exemplary alkyl groups include —SCH3 and the like.
The term “halo” or “halogen” as used herein by itself or part of another group refers to fluoro, chloro, bromo or iodo. In one embodiment, the halo is fluoro or chloro.
The term “amino” as used herein by itself or part of another group refers to a radical of formula —NRaRb wherein Ra and Rb are independently hydrogen, haloalkyl, aralkyl, optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted heterocyclo, optionally substituted aryl or optionally substituted heteroaryl; or Ra and Rb taken together with the nitrogen atom to which they are attached form a four to seven membered optionally substituted heterocyclo. Exemplary amino groups include —NH2, —N(H)CH3, —N(CH3)2, N(H)CH2CH3, N(CH2CH3), —N(H)CH2Ph and the like.
The term “carboxamido” as used herein by itself or part of another group refers to a radical of formula —CO-amino. Exemplary carboxamido groups include —CONH2, —CON(H)CH3, —CON(H)Ph, —CON(H)CH2CH2Ph, —CON(CH3)2, CON(H)CHPh2 and the like.
The term “sulfonamido” as used herein by itself or part of another group refers to a radical of formula —SO2-amino. Exemplary sulfonamido groups include —SO2NH2, —SO2N(H)CH3, —SO2N(H)Ph and the like.
The term “about,” as used herein, includes the recited number ±10%. Thus, “about 10” means 9 to 11.
The term “leaving group” as used herein refers to an atom or group that becomes detached from an atom or group in what is considered to be the residual or main part of the substrate in a specified reaction. In amide coupling reactions, exemplary leaving groups (i.e., leaving groups designated L1) include —F, —Cl, —Br, —OH, —OC6F5, —O(CO)alkyl and the like. In one embodiment, the leaving group, L1, is —Cl. In another embodiment, the leaving group, L1, is an activated form of —OH (e.g., OBt, O-acylisourea). An activating agent (e.g., dicyclohexylcarbodiimide (DCC), 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC), benzotriazol-1-yloxy)tripyrrolidinophosphonium hexafluorophosphate (PyBop)) may be employed to active a carboxylic acid (i.e, the leaving group is —OH) toward amide formation. Such activating agents are well known to those of skill in the art of organic synthesis. Other additives, such as N-hydroxybenzotriazole (HOBt) or N-hydroxysuccinimide (HOSu), may also be added to optimize reaction parameters (e.g., rate, yield, purity, racemization). In nucleophilic displacement reactions (e.g., SN2 reactions), exemplary leaving groups (i.e., leaving groups designated L2) include —Cl, —Br, —I, —OSO2Me (mesylate), —OSO2CF3 (triflate), —OSO2C6H5 (besylate), —OSO2CH3C6H4 (tosylate) and the like. In one embodiment, the leaving group, L2, is —Cl or —Br. In another embodiment, the leaving group, L2, is —Br.
The term “amine protecting group” as used herein refers to group that blocks (i.e., protects) the amine functionality while reactions are carried out on other functional groups or parts of the molecule. Those skilled in the art will be familiar with the selection, attachment, and cleavage of amine protecting groups and will appreciate that many different protective groups are known in the art, the suitability of one protective group or another being dependent on the particular the synthetic scheme planned. Treatises on the subject are available for consultation, such as Greene and Wuts, “Protective Groups in Organic Synthesis,” 3rd Ed., pp. 17-245 (J. Wiley & Sons, 1999), the disclosure of which is incorporated by reference. Suitable amine protecting groups include the carbobenzyloxy (Cbz), tert-butyloxycarbonyl (BOC), 9-fluorenylmethyloxycarbonyl (FMOC) and benzyl (Bn) group.
Throughout the specification, groups an optional substituents thereof are chosen to provide stable moieties and compounds.
Certain of the compounds of the present invention may exist as stereoisomers including optical isomers. The invention includes all stereoisomers, both as pure individual stereoisomer preparations and enriched preparations of each, and both the racemic mixtures of such stereoisomers as well as the individual enantiomers that may be separated according to methods that are well known to those of skill in the art.
In certain embodiments of the invention, the compound of Formula I is:










In other embodiments of the invention, the compound of Formula I is:













In particular embodiments of the invention the compound of Formula I is selected from the group consisting of:










or a pharmaceutically acceptable salt or prodrug thereof.

The present invention also pertains to a process for the preparation of a compound of Formula XII




comprising:

a) condensing a compound of Formula XIII





with ammonia to give a compound of Formula XIV





b) converting a compound of Formula XIV to a compound of Formula XV





c) condensing a compound of Formula XV with a compound of Formula XVI





wherein L2 is a leaving group, to give a compound of Formula XVII,





and b) cyclizing a compound of Formula XVII, to give a compound of Formula XII, wherein:

R13 is selected from the group consisting of N(H)P1 and





P1 is an amine protecting group;

A1 and A2 are independently selected from the group consisting of hydrogen and optionally substituted alkyl, wherein A2 is absent when V is O;

V is selected from the group consisting of N, CH and O;

W is selected from the group consisting of CH and N;

X is selected from the group consisting of hydrogen, optionally substituted alkyl and aralkyl;

Y is selected from the group consisting of CON(R1), N(R1)CO, C(O)O, OC(O), (CH2)1-3, wherein one or more CH2 groups can be replaced by O, S, or NR1, optionally substituted aryl and optionally substituted heteroaryl;

Z is (CR2aR2b)r;

U is selected from the group consisting of CR5aR5b and NR6;

m is 1 or 2;

r is 0-3;

R1 is selected from the group consisting of hydrogen, optionally substituted alkyl, aralkyl, optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heteroaryl and optionally substituted heterocyclo;

each R2a, R2b, R5a and R5b are independently selected from the group consisting of hydrogen, optionally substituted alkyl, aralkyl, optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heteroaryl and optionally substituted heterocyclo;

R6 is selected from the group consisting of hydrogen, optionally substituted alkyl, aralkyl, optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heteroaryl and optionally substituted heterocyclo and COR9;

R9 is selected from the group consisting of hydrogen, optionally substituted alkyl, aralkyl, optionally substituted cycloalkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted aryl, optionally substituted heteroaryl and optionally substituted heterocyclo; and

R10a and R10b are independently selected from the group consisting of hydrogen, optionally substituted alkyl, haloalkyl, aralkyl, optionally substituted cycloalkyl, optionally substituted cycloalkenyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted aryl, optionally substituted heteroaryl and optionally substituted heterocyclo.

In one embodiment, R13 is —N(H)P1. In one embodiment, P1 is selected from the group consisting of carbobenzyloxy, tert-butyloxycarbonyl and benzyl. In one embodiment, P1 is selected from the group consisting of carbobenzyloxy and tert-butyloxycarbonyl. In one embodiment, P1 is carbobenzyloxy. In one embodiment, P1 is tert-butyloxycarbonyl. In one embodiment, U is CH2. In one embodiment, U is NCOR9. In another embodiment, R9 is optionally substituted alkyl. In one embodiment, m is 2. In one embodiment, m is 1.
In one embodiment, L2 is selected from the group consisting of Cl, —Br, —I, —OSO2Me, —OSO2CF3, —OSO2C6H5 and —OSO2CH3C6H4. In another embodiment, L2 is selected from the group consisting of —Cl and —Br. In another embodiment, L2 is Br. In one embodiment, R10b is optionally substituted phenyl. In one embodiment, R10b is hydrogen.
In one embodiment, R13 is




In one embodiment, Y is CON(H),

W is CH, r is 0 and V is N. In one embodiment, U is CH2. In one embodiment, U is NCOR9. In another embodiment, R9 is optionally substituted alkyl. In one embodiment, m is 2. In one embodiment, m is 1.
In one embodiment, ammonia is condensed with a compound of Formula XIII in the presence of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide and N-hydroxybenzotriazole in an inert organic solvent (e.g., acetonitrile, tetrahydrofuran, dimethylformamide, etc.) at a temperature of about −20° C. to about 25° C. In one embodiment, a compound of XIV is converted to a compound of Formula XV using P4S10 in an inert organic solvent (e.g., dichloromethane, etc.) at a temperature of about −20° C. to about 45° C. In one embodiment, the condensation of a compound of Formula XV with a compound of Formula XVI is carried out in an inert organic solvent (e.g., acetonitrile, tetrahydrofuran, dimethylformamide, etc.) at a temperature of about 0° C. to about 50° C. In one embodiment, a compound of Formula XVII is cyclized in methanol, ethanol, propanol, isopropanol, or butanol. In one embodiment, the cyclization of a compound of Formula XVII is carried out at a temperature of about 25° C. to about 100° C., in one embodiment, above 30° C. In one embodiment, the cyclization of a compound of Formula XVIII is carried out in refluxing solvent. In one embodiment, the cyclization of a compound of Formula XVIII is carried out in refluxing ethanol.
The progress of any of the above reactions can be monitored by analytical methods known in the art such as TLC, LC, LC/MS, HPLC, NMR, etc. A compound of Formula XII, as well as any synthetic intermediates (i.e., a compound of Formula XIV, XV or XVII), can be isolated and purified by any means known in the art such normal- and reverse-phase column chromatography (e.g., column chromatography on silica gel or reverse-phase HPLC), crystallization, extraction, etc. The product thus isolated can be subjected to further purification (e.g., recrystallization) until the desired level of purity is achieved. In one embodiment, a compound of Formula XII has a purity of 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more.
In another embodiment, the invention pertains to a process for the preparation of a compound of Formula XVIII




comprising:

a) condensing a compound of Formula XIII





with a compound of Formula XIX





to give a compound of Formula XX





and b) cyclizing a compound of Formula XX, to give a compound of Formula XVIII, wherein:

R13 is selected from the group consisting of N(H)P1 and





P1 is an amine protecting group;

A1 and A2 are independently selected from the group consisting of hydrogen and optionally substituted alkyl, wherein A2 is absent when V is O;

V is selected from the group consisting of N, CH and O;

W is selected from the group consisting of CH and N;

X is selected from the group consisting of hydrogen, optionally substituted alkyl and aralkyl;

Y is selected from the group consisting of CON(R1), N(R1)CO, C(O)O, OC(O), (CH2)1-3, wherein one or more CH2 groups can be replaced by O, S, or NR1, optionally substituted aryl and optionally substituted heteroaryl;

Z is (CR2aR2b)r;

U is selected from the group consisting of CR5aR5b and NR6;

m is 1 or 2;

r is 0-3;

R1 is selected from the group consisting of hydrogen, optionally substituted alkyl, aralkyl, optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heteroaryl and optionally substituted heterocyclo;

each R2a, R2b, R5a and R5b are independently selected from the group consisting of hydrogen, optionally substituted alkyl, aralkyl, optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heteroaryl and optionally substituted heterocyclo;

R6 is selected from the group consisting of hydrogen, optionally substituted alkyl, aralkyl, optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heteroaryl and optionally substituted heterocyclo and COR9;

R9 is selected from the group consisting of hydrogen, optionally substituted alkyl, aralkyl, optionally substituted cycloalkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted aryl, optionally substituted heteroaryl and optionally substituted heterocyclo;

R11a, R11b, R11c and R11d are independently selected from the group consisting of hydrogen, optionally substituted alkyl, haloalkyl, aralkyl, optionally substituted cycloalkyl, optionally substituted cycloalkenyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocyclo, halo, nitro, cyano, hydroxy, amino, alkoxy, aryloxy, arylalkyloxy, alkylthio, carboxamido and sulfonamido, and

R12 is selected from the group consisting of hydrogen, optionally substituted alkyl, aralkyl, optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heteroaryl and optionally substituted heterocyclo.

In one embodiment, R11a is optionally substituted phenyl. In one embodiment, R11b, R11c and R11d are hydrogen. In one embodiment, R11a is optionally substituted phenyl, and R11b, R11c and R11d are hydrogen. In one embodiment, m is 2. In one embodiment, m is 1. In one embodiment, U is CH2. In one embodiment, U is NCOR9. In another embodiment, R9 is optionally substituted alkyl. In one embodiment, R12 is hydrogen.
In one embodiment, R13 is —N(H)P1. In one embodiment, P1 is selected from the group consisting of carbobenzyloxy, tert-butyloxycarbonyl and benzyl. In one embodiment, P1 is selected from the group consisting of carbobenzyloxy and tert-butyloxycarbonyl. In one embodiment, P1 is carbobenzyloxy. In one embodiment, P1 is tert-butyloxycarbonyl. In one embodiment, U is CH2. In one embodiment, U is NCOR9. In another embodiment, R9 is optionally substituted alkyl. In one embodiment, m is 2. In one embodiment, m is 1.
In one embodiment, R13 is




In one embodiment, Y is CON(H), W is CH, r is 0 and V is N. In one embodiment, U is CH2. In one embodiment, U is NCOR9. In another embodiment, R9 is optionally substituted alkyl. In one embodiment, m is 2. In one embodiment, m is 1.

In one embodiment, a compound of Formula XIII is condensed with a compound of Formula XIX in the presence of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide and N-hydroxybenzotriazole in an inert organic solvent (e.g., acetonitrile, tetrahydrofuran, dimethylformamide, etc.) at a temperature of about −20° C. to about 25° C. In one embodiment, a compound of Formula XX is cyclized in acetic acid at a temperature of about 25° C. to about 118° C. In another embodiment, a compound of Formula XX is cyclized in refluxing acetic acid.
The progress of any of the above reactions can be monitored by analytical methods known in the art such as TLC, LC, LC/MS, HPLC, NMR, etc. A compound of Formula XVIII, as well as any synthetic intermediate(s) (i.e., a compound of XIX), can be isolated and purified by any means known in the art such normal- and reverse-phase column chromatography (e.g., column chromatography on silica gel or reverse-phase HPLC), crystallization, extraction, etc. The product thus isolated can be subjected to further purification (e.g., recrystallization) until the desired level of purity is achieved. In one embodiment, a compound of Formula XVIII has a purity of 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more.
The present invention also pertains to a process for preparing a compound of Formula XXI




comprising:

condensing a compound of Formula XXIII





with a compound of Formula XXIV




wherein L1 is a leaving group, to give a compound of Formula XXI,

wherein:

A1 and A2 are independently selected from the group consisting of hydrogen and optionally substituted alkyl, wherein A2 is absent when V is O;

V is selected from the group consisting of N, CH and O;

W is selected from the group consisting of CH and N;

X is selected from the group consisting of hydrogen, optionally substituted alkyl and aralkyl;

Z is (CR2aR2b)r;

U is selected from the group consisting of CR5aR5b and NR6;

m is 1 or 2;

r is 0-3;

each R2a, R2b, R5a and R5b are independently selected from the group consisting of hydrogen, optionally substituted alkyl, aralkyl, optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heteroaryl and optionally substituted heterocyclo;

R6 is selected from the group consisting of hydrogen, optionally substituted alkyl, aralkyl, optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heteroaryl and optionally substituted heterocyclo and COR9;

R9 is selected from the group consisting of hydrogen, optionally substituted alkyl, aralkyl, optionally substituted cycloalkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted aryl, optionally substituted heteroaryl and optionally substituted heterocyclo; and

T is optionally substituted heteroaryl.

In one embodiment, W is CH and V is N. In one embodiment, T is selected from the group consisting of




wherein:

Q is selected from the group consisting of O, S and NR12;

R12 is selected from the group consisting of hydrogen, optionally substituted alkyl, aralkyl, optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heteroaryl and optionally substituted heterocyclo; and

R10a, R10b, R11a, R11b, R11c and R11d are independently selected from the group consisting of hydrogen, optionally substituted alkyl, haloalkyl, aralkyl, optionally substituted cycloalkyl, optionally substituted cycloalkenyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocyclo, halo, nitro, cyano, hydroxy, amino, alkoxy, aryloxy, arylalkyloxy, alkylthio, carboxamido and sulfonamido. In one embodiment, U is CH2. In one embodiment, U is NCOR9. In one embodiment, R9 is optionally substituted alkyl. In one embodiment, m is 2. In one embodiment, m is 1.

In one embodiment, L1 is selected from the group consisting of —Cl and —OH. In one embodiment, L1 is —OH and the reaction is carried out in the presence of an activating agent.
In one embodiment, a compound of Formula XXIII is prepared by removing P1 from a compound of Formula XXII




wherein P1 is a leaving group. In one embodiment, P1 is carbobenzyloxy, tert-butyloxycarbonyl, 9-fluorenylmethyloxycarbonyl or benzyl. In one embodiment, P1 is carbobenzyloxy or tert-butyloxycarbonyl. In one embodiment, P1 is carbobenzyloxy.

In one embodiment, the condensation reaction is or tert-butyloxycarbonyl. In one embodiment, the condensation reaction is conducted in an inert organic solvent such as acetonitrile, benzene, chloroform, 1,2-dichloroethane, 1,2-dimethoxyethane, dimethylformamide, dimethylsulfoxide, dioxane, dichloromethane, N-methyl-2-pyrrolidinone or tetrahydrofuran. In another embodiment, the condensation reaction is carried out in tetrahydrofuran. In another embodiment, the condensation reaction is carried out in dichloromethane. In one embodiment, the condensation reaction is carried out at about −20° C. to about 35° C. In another embodiment, the condensation reaction is carried out at about 25° C. In one embodiment, the condensation reaction is complete in about 1 hour to about 48 hours. In another embodiment, the condensation reaction is complete in about 12 hours.
In one embodiment, L1 is Cl, —OH or —OBt. In one embodiment, L1 is —OH or —OBt. In another embodiment, the condensation reaction is carried out in the presence of an activating agent. In another embodiment, the activating agent is dicyclohexylcarbodiimide, 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide or benzotriazol-1-yloxy)tripyrrolidinophosphonium hexafluorophosphate. In another embodiment, the activating agent is 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide. In another embodiment, the condensation reaction is carried out in the presence of an activating agent and an additive that optimize reaction parameters such as purity and yield. In another embodiment, the additive is N-hydroxybenzotriazole.
The progress of the condensation reaction between a compound of Formula XXIII and Formula XXIV can be monitored by analytical methods known in the art such as TLC, LC, LC/MS, HPLC, NMR, etc. A compound of Formula XXI can be isolated and purified by any means known in the art such normal- and reverse-phase column chromatography (e.g., column chromatography on silica gel or reverse-phase HPLC), crystallization, extraction, etc. The product thus isolated can be subjected to further purification (e.g., recrystallization) until the desired level of purity is achieved. In one embodiment, a compound of Formula XXI has a purity of 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more.
In another embodiment, the invention pertains to a compound having Formula XXII:




wherein:

T is optionally substituted heteroaryl;

m is 1 or 2;

U is CH2 or NR6 

R6 is selected from the group consisting of hydrogen, optionally substituted alkyl, aralkyl, optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heteroaryl and optionally substituted heterocyclo and COR9;

R9 is selected from the group consisting of hydrogen, optionally substituted alkyl, aralkyl, optionally substituted cycloalkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted aryl, optionally substituted heteroaryl and optionally substituted heterocyclo; and

P1 is an amine protecting group.

In one embodiment, P1 is selected from the group consisting of t-butoxycarbonyl and benzyloxycarbonyl.
In another embodiment, the invention pertains to a compound having Formula XXV:




wherein:

T is optionally substituted heteroaryl;

m is 1 or 2;

U is CH2 or NR6 

R6 is selected from the group consisting of hydrogen, optionally substituted alkyl, aralkyl, optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heteroaryl and optionally substituted heterocyclo and COR9;

R9 is selected from the group consisting of hydrogen, optionally substituted alkyl, aralkyl, optionally substituted cycloalkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted aryl, optionally substituted heteroaryl and optionally substituted heterocyclo;

X is selected from the group consisting of hydrogen, optionally substituted alkyl, and aralkyl;

A1 is selected from the group consisting of hydrogen an optionally substituted alkyl; and

P1 is an amine protecting group.

In one embodiment, P1 is selected from the group consisting of t-butoxycarbonyl and benzyloxycarbonyl.
In one embodiment the compound of Formula XXV is:




The compounds of this invention may be prepared using methods known to those of skill in the art. Specifically, compounds of the invention can be prepared as illustrated by the exemplary reactions in the Examples.
An important aspect of the present invention is that compounds of Formulae I-XIa induce apoptosis and also potentiate the induction of apoptosis in response to apoptosis induction signals. Therefore, it is contemplated that these compounds will sensitize cells to inducers of apoptosis, including cells that are resistant to such inducers. The IAP inhibitors of the present invention can be used to induce apoptosis in any disorder that can be treated, ameliorated, or prevented by the induction of apoptosis. Thus, the present invention provides compositions and methods for targeting animals characterized as overexpressing an IAP protein. In some of the embodiments, the cells (e.g., cancer cells) will show elevated expression levels of IAP proteins as compared to non-pathological samples (e.g., non-cancerous cells). In other embodiments, the cells operationally will manifest elevated expression levels of IAP proteins by virtue of executing the apoptosis program and dying in response to an inhibiting effective amount of a compound of Formulae I-XIa, said response occurring, at least in part, due to the dependence in such cells on IAP protein function for their survival.
In another embodiment, the invention pertains to modulating an apoptosis-associated state which is associated with one or more apoptosis-modulating agents. Examples of apoptosis-modulating agents include, but are not limited to, Fas/CD95, TRAMP, TNF RI, DR1, DR2, DR3, DR4, DR5, DR6, FADD, RIP, TNFα, Fas ligand, TRAIL, antibodies to TRAIL-R1 or TRAIL-R2, Bcl-2, p53, BAX, BAD, Akt, CAD, PI3 kinase, PP1, and caspase proteins. Other agents involved in the initiation, decision and degradation phase of apoptosis are also included. Examples of apoptosis-modulating agents include agents, the activity, presence, or change in concentration of which, can modulate apoptosis in a subject. Preferred apoptosis-modulating agents are inducers of apoptosis, such as TNF or a TNF-related ligand, particularly a TRAMP ligand, a Fas/CD95 ligand, a TNFR-1 ligand, or TRAIL.
In some embodiments, the compositions and methods of the present invention are used to treat diseased cells, tissues, organs, or pathological conditions and/or disease states in an animal (e.g., a mammalian subject including, but not limited to, humans and veterinary animals). In this regard, various diseases and pathologies are amenable to treatment or prophylaxis using the present methods and compositions. A non-limiting exemplary list of these diseases and conditions includes, but is not limited to, breast cancer, prostate cancer, lymphoma, skin cancer, pancreatic cancer, colon cancer, melanoma, malignant melanoma, ovarian cancer, brain cancer, primary brain carcinoma, head-neck cancer, glioma, glioblastoma, liver cancer, bladder cancer, non-small cell lung cancer, head or neck carcinoma, breast carcinoma, ovarian carcinoma, lung carcinoma, small-cell lung carcinoma, Wilms' tumor, cervical carcinoma, testicular carcinoma, bladder carcinoma, pancreatic carcinoma, stomach carcinoma, colon carcinoma, prostatic carcinoma, genitourinary carcinoma, thyroid carcinoma, esophageal carcinoma, myeloma, multiple myeloma, adrenal carcinoma, renal cell carcinoma, endometrial carcinoma, adrenal cortex carcinoma, malignant pancreatic insulinoma, malignant carcinoid carcinoma, choriocarcinoma, mycosis fungoides, malignant hypercalcemia, cervical hyperplasia, leukemia, acute lymphocytic leukemia, chronic lymphocytic leukemia, acute myelogenous leukemia, chronic myelogenous leukemia, chronic granulocytic leukemia, acute granulocytic leukemia, hairy cell leukemia, neuroblastoma, rhabdomyosarcoma, Kaposi's sarcoma, polycythemia vera, essential thrombocytosis, Hodgkin's disease, non-Hodgkin's lymphoma, soft-tissue sarcoma, osteogenic sarcoma, primary macroglobulinemia, and retinoblastoma, and the like, T and B cell mediated autoimmune diseases; inflammatory diseases; infections (e.g., as antiulcerous agents, e.g., in the context of H. pylori infection); hyperproliferative diseases; AIDS; degenerative conditions; vascular diseases (e.g., primary varicosis), and the like. The compounds of the present invention may also be useful in the treatment of diseases in which there is a defect in the programmed cell-death or the apoptotic machinery e.g., multiple sclerosis, asthma, artherosclerosis and the like. In some embodiments, the cancer cells being treated are metastatic. In other embodiments, the cancer cells being treated are resistant to anticancer agents.
In some embodiments, infections suitable for treatment with the compositions and methods of the present invention include, but are not limited to, infections caused by viruses, bacteria, fungi, mycoplasma, prions, and the like.
Some embodiments of the present invention provide methods for administering an effective amount of a compound of Formulae I-XIa and at least one additional therapeutic agent (including, but not limited to, chemotherapeutic antineoplastics, apoptosis-modulating agents, antimicrobials, antivirals, antifungals, and anti-inflammatory agents) and/or therapeutic technique (e.g., surgical intervention, and/or radiotherapies).
A number of suitable anticancer agents are contemplated for use in the methods of the present invention. Indeed, the present invention contemplates, but is not limited to, administration of numerous anticancer agents such as: agents that induce apoptosis; polynucleotides (e.g., anti-sense, ribozymes, siRNA); polypeptides (e.g., enzymes and antibodies); biological mimetics (e.g., gossypol or BH3 mimetics); agents that bind (e.g., oligomerize or complex) with a Bcl-2 family protein such as Bax; alkaloids; alkylating agents; antitumor antibiotics; antimetabolites; hormones; platinum compounds; monoclonal or polyclonal antibodies (e.g., antibodies conjugated with anticancer drugs, toxins, defensins), toxins; radionuclides; biological response modifiers (e.g., interferons (e.g., IFN-α) and interleukins (e.g., IL-2)); adoptive immunotherapy agents; hematopoietic growth factors; agents that induce tumor cell differentiation (e.g., all-trans-retinoic acid); gene therapy reagents (e.g., antisense therapy reagents and nucleotides); tumor vaccines; angiogenesis inhibitors; proteosome inhibitors: NF-KB modulators; anti-CDK compounds; HDAC inhibitors; and the like. Numerous other examples of chemotherapeutic compounds and anticancer therapies suitable for co-administration with the disclosed compounds are known to those skilled in the art.
In certain embodiments, anticancer agents comprise agents that induce or stimulate apoptosis. Agents that induce apoptosis include, but are not limited to, radiation (e.g., X-rays, gamma rays, UV); tumor necrosis factor (TNF)-related factors (e.g., TNF family receptor proteins, TNF family ligands, TRAIL, antibodies to TRAIL-R1 or TRAIL-R2); kinase inhibitors (e.g., epidermal growth factor receptor (EGFR) kinase inhibitor, vascular growth factor receptor (VGFR) kinase inhibitor, fibroblast growth factor receptor (FGFR) kinase inhibitor, platelet-derived growth factor receptor (PDGFR) kinase inhibitor, and Bcr-Abl kinase inhibitors (such as GLEEVEC)); antisense molecules; antibodies (e.g., HERCEPTIN, RITUXAN, ZEVALIN, and AVASTIN); anti-estrogens (e.g., raloxifene and tamoxifen); anti-androgens (e.g., flutamide, bicalutamide, finasteride, aminoglutethamide, ketoconazole, and corticosteroids); cyclooxygenase 2 (COX-2) inhibitors (e.g., celecoxib, meloxicam, NS-398, and non-steroidal anti-inflammatory drugs (NSAIDs)); anti-inflammatory drugs (e.g., butazolidin, DECADRON, DELTASONE, dexamethasone, dexamethasone intensol, DEXONE, HEXADROL, hydroxychloroquine, METICORTEN, ORADEXON, ORASONE, oxyphenbutazone, PEDIAPRED, phenylbutazone, PLAQUENIL, prednisolone, prednisone, PRELONE, and TANDEARIL); and cancer chemotherapeutic drugs (e.g., irinotecan (CAMPTOSAR), CPT-11, fludarabine (FLUDARA), dacarbazine (DTIC), dexamethasone, mitoxantrone, MYLOTARG, VP-16, cisplatin, carboplatin, oxaliplatin, 5-FU, doxorubicin, gemcitabine, bortezomib, gefitinib, bevacizumab, TAXOTERE or TAXOL); cellular signaling molecules; ceramides and cytokines; staurosporine, and the like.
In still other embodiments, the compositions and methods of the present invention provide a compound of Formulae I-XIa and at least one anti-hyperproliferative or antineoplastic agent selected from alkylating agents, antimetabolites, and natural products (e.g., herbs and other plant and/or animal derived compounds).
Alkylating agents suitable for use in the present compositions and methods include, but are not limited to: 1) nitrogen mustards (e.g., mechlorethamine, cyclophosphamide, ifosfamide, melphalan (L-sarcolysin); and chlorambucil); 2) ethylenimines and methylmelamines (e.g., hexamethylmelamine and thiotepa); 3) alkyl sulfonates (e.g., busulfan); 4) nitrosoureas (e.g., carmustine (BCNU); lomustine (CCNU); semustine (methyl-CCNU); and streptozocin (streptozotocin)); and 5) triazenes (e.g., dacarbazine (DTIC; dimethyltriazenoimid-azolecarboxamide).
In some embodiments, antimetabolites suitable for use in the present compositions and methods include, but are not limited to: 1) folic acid analogs (e.g., methotrexate (amethopterin)); 2) pyrimidine analogs (e.g., fluorouracil (5-fluorouracil; 5-FU), floxuridine (fluorode-oxyuridine; FudR), and cytarabine (cytosine arabinoside)); and 3) purine analogs (e.g., mercaptopurine (6-mercaptopurine; 6-MP), thioguanine (6-thioguanine; TG), and pentostatin (2′-deoxycoformycin)).
In still further embodiments, chemotherapeutic agents suitable for use in the compositions and methods of the present invention include, but are not limited to: 1) vinca alkaloids (e.g., vinblastine (VLB), vincristine); 2) epipodophyllotoxins (e.g., etoposide and teniposide); 3) antibiotics (e.g., dactinomycin (actinomycin D), daunorubicin (daunomycin; rubidomycin), doxorubicin, bleomycin, plicamycin (mithramycin), and mitomycin (mitomycin C)); 4) enzymes (e.g., L-asparaginase); 5) biological response modifiers (e.g., interferon-alfa); 6) platinum coordinating complexes (e.g., cisplatin (cis-DDP) and carboplatin); 7) anthracenediones (e.g., mitoxantrone); 8) substituted ureas (e.g., hydroxyurea); 9) methylhydrazine derivatives (e.g., procarbazine (N-methylhydrazine; M1H)); 10) adrenocortical suppressants (e.g., mitotane (o,p′-DDD) and aminoglutethimide); 11) adrenocorticosteroids (e.g., prednisone); 12) progestins (e.g., hydroxyprogesterone caproate, medroxyprogesterone acetate, and megestrol acetate); 13) estrogens (e.g., diethylstilbestrol and ethinyl estradiol); 14) antiestrogens (e.g., tamoxifen); 15) androgens (e.g., testosterone propionate and fluoxymesterone); 16) antiandrogens (e.g., flutamide): and 17) gonadotropin-releasing hormone analogs (e.g., leuprolide).
Any oncolytic agent that is routinely used in a cancer therapy context finds use in the compositions and methods of the present invention. For example, the U.S. Food and Drug Administration maintains a formulary of oncolytic agents approved for use in the United States. International counterpart agencies to the U.S.F.D.A. maintain similar formularies. Table 1 provides a list of exemplary antineoplastic agents approved for use in the U.S. Those skilled in the art will appreciate that the “product labels” required on all U.S. approved chemotherapeutics describe approved indications, dosing information, toxicity data, and the like, for the exemplary agents.









TABLE 1







Aldesleukin
Proleukin
Chiron Corp., Emeryville, CA


(des-alanyl-1, serine-125 human interleukin-2)


Alemtuzumab
Campath
Millennium and ILEX


(IgG1κ anti CD52 antibody)

Partners, LP, Cambridge, MA


Alitretinoin
Panretin
Ligand Pharmaceuticals, Inc.,


(9-cis-retinoic acid)

San Diego CA


Allopurinol
Zyloprim
GlaxoSmithKline, Research


(1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one

Triangle Park, NC


monosodium salt)


Altretamine
Hexalen
US Bioscience, West


(N,N,N′,N′,N′′,N′′,-hexamethyl-1,3,5-triazine-2,4,6-

Conshohocken, PA


triamine)


Amifostine
Ethyol
US Bioscience


(ethanethiol, 2-[(3-aminopropyl)amino]-, dihydrogen


phosphate (ester))


Anastrozole
Arimidex
AstraZeneca Pharmaceuticals,


(1,3-Benzenediacetonitrile,a,a,a′,a′-tetramethyl-5-(1H-

LP, Wilmington, DE


1,2,4-triazol-1-ylmethyl))


Arsenic trioxide
Trisenox
Cell Therapeutic, Inc., Seattle,




WA


Asparaginase
Elspar
Merck & Co., Inc.,


(L-asparagine amidohydrolase, type EC-2)

Whitehouse Station, NJ


BCG Live
TICE BCG
Organon Teknika, Corp.,


(lyophilized preparation of an attenuated strain of

Durham, NC


Mycobacterium bovis (Bacillus Calmette-Gukin [BCG],


substrain Montreal)


bexarotene capsules
Targretin
Ligand Pharmaceuticals


(4-[1-(5,6,7,8-tetrahydro-3,5,5,8,8-pentamethyl-2-


napthalenyl) ethenyl] benzoic acid)


bexarotene gel
Targretin
Ligand Pharmaceuticals


Bleomycin
Blenoxane
Bristol-Myers Squibb Co.,


(cytotoxic glycopeptide antibiotics produced by

NY, NY


Streptomyces verticillus; bleomycin A2 and bleomycin


B2)


Capecitabine
Xeloda
Roche


(5′-deoxy-5-fluoro-N-[(pentyloxy)carbonyl]-cytidine)


Carboplatin
Paraplatin
Bristol-Myers Squibb


(platinum, diammine [1,1-cyclobutanedicarboxylato(2-)-


0,0′]-,(SP-4-2))


Carmustine
BCNU,
Bristol-Myers Squibb


(1,3-bis(2-chloroethyl)-1-nitrosourea)
BiCNU


Carmustine with Polifeprosan 20 Implant
Gliadel Wafer
Guilford Pharmaceuticals,




Inc., Baltimore, MD


Celecoxib
Celebrex
Searle Pharmaceuticals,


(as 4-[5-(4-methylphenyl)-3-(trifluoromethyl)-1H-

England


pyrazol-1-yl] benzenesulfonamide)


Chlorambucil
Leukeran
GlaxoSmithKline


(4-[bis(2chlorethyl)amino]benzenebutanoic acid)


Cisplatin
Platinol
Bristol-Myers Squibb


(PtCl2H6N2)


Cladribine
Leustatin, 2-
R. W. Johnson Pharmaceutical


(2-chloro-2′-deoxy-b-D-adenosine)
CdA
Research Institute, Raritan, NJ


Cyclophosphamide
Cytoxan,
Bristol-Myers Squibb


(2-[bis(2-chloroethyl)amino] tetrahydro-2H-13,2-
Neosar


oxazaphosphorine 2-oxide monohydrate)


Cytarabine
Cytosar-U
Pharmacia & Upjohn


(1-b-D-Arabinofuranosylcytosine, C9H13N3O5)

Company


cytarabine liposomal
DepoCyt
Skye Pharmaceuticals, Inc.,




San Diego, CA


Dacarbazine
DTIC-Dome
Bayer AG, Leverkusen,


(5-(3,3-dimethyl-1-triazeno)-imidazole-4-carboxamide

Germany


(DTIC))


Dactinomycin, actinomycin D
Cosmegen
Merck


(actinomycin produced by Streptomyces parvullus,


C62H86N12O16)


Darbepoetin alfa
Aranesp
Amgen, Inc., Thousand Oaks,


(recombinant peptide)

CA


daunorubicin liposomal
DanuoXome
Nexstar Pharmaceuticals, Inc.,


((8S-cis)-8-acetyl-10-[(3-amino-2,3,6-trideoxy-á-L-lyxo-

Boulder, CO


hexopyranosyl)oxy]-7,8,9,10-tetrahydro-6,8,11-


trihydroxy-1-methoxy-5,12-naphthacenedione


hydrochloride)


Daunorubicin HCl, daunomycin
Cerubidine
Wyeth Ayerst, Madison, NJ


((1S,3S)-3-Acetyl-1,2,3,4,6,11-hexahydro-3,5,12-


trihydroxy-10-methoxy-6,11-dioxo-1-naphthacenyl 3-


amino-2,3,6-trideoxy-(alpha)-L-lyxo-hexopyranoside


hydrochloride)


Denileukin diftitox
Ontak
Seragen, Inc., Hopkinton, MA


(recombinant peptide)


Dexrazoxane
Zinecard
Pharmacia & Upjohn


((S)-4,4′-(1-methyl-1,2-ethanediyl)bis-2,6-

Company


piperazinedione)


Docetaxel
Taxotere
Aventis Pharmaceuticals, Inc.,


((2R,3S)-N-carboxy-3-phenylisoserine, N-tert-butyl ester,

Bridgewater, NJ


13-ester with 5b-20-epoxy-12a,4,7b,10b,13a-


hexahydroxytax-11-en-9-one 4-acetate 2-benzoate,


trihydrate)


Doxorubicin HCl
Adriamycin,
Pharmacia & Upjohn


(8S,10S)-10-[(3-amino-2,3,6-trideoxy-a-L-lyxo-
Rubex
Company


hexopyranosyl)oxy]-8-glycolyl-7,8,9,10-tetrahydro-


6,8,11-trihydroxy-1-methoxy-5,12-naphthacenedione


hydrochloride)


doxorubicin
Adriamycin
Pharmacia & Upjohn



PFS
Company



Intravenous



injection


doxorubicin liposomal
Doxil
Sequus Pharmaceuticals, Inc.,




Menlo park, CA


dromostanolone propionate
Dromostanolone
Eli Lilly & Company,


(17b-Hydroxy-2a-methyl-5a-androstan-3-one propionate)

Indianapolis, IN


dromostanolone propionate
Masterone
Syntex, Corp., Palo Alto, CA



injection


Elliott's B Solution
Elliott's B
Orphan Medical, Inc



Solution


Epirubicin
Ellence
Pharmacia & Upjohn


((8S-cis)-10-[(3-amino-2,3,6-trideoxy-a-L-arabino-

Company


hexopyranosyl)oxy]-7,8,9,10-tetrahydro-6,8,11-


trihydroxy-8-(hydroxyacetyl)-1-methoxy-5,12-


naphthacenedione hydrochloride)


Epoetin alfa
Epogen
Amgen, Inc


(recombinant peptide)


Estramustine
Emcyt
Pharmacia & Upjohn


(estra-1,3,5(10)-triene-3,17-diol(17(beta))-, 3-[bis(2-

Company


chloroethyl)carbamate] 17-(dihydrogen phosphate),


disodium salt, monohydrate, or estradiol 3-[bis(2-


chloroethyl)carbamate] 17-(dihydrogen phosphate),


disodium salt, monohydrate)


Etoposide phosphate
Etopophos
Bristol-Myers Squibb


(4′-Demethylepipodophyllotoxin 9-[4,6-O-(R)-


ethylidene-(beta)-D-glucopyranoside], 4′-(dihydrogen


phosphate))


etoposide, VP-16
Vepesid
Bristol-Myers Squibb


(4′-demethylepipodophyllotoxin 9-[4,6-0-(R)-ethylidene-


(beta)-D-glucopyranoside])


Exemestane
Aromasin
Pharmacia & Upjohn


(6-methylenandrosta-1,4-diene-3,17-dione)

Company


Filgrastim
Neupogen
Amgen, Inc


(r-metHuG-CSF)


floxuridine (intraarterial)
FUDR
Roche


(2′-deoxy-5-fluorouridine)


Fludarabine
Fludara
Berlex Laboratories, Inc.,


(fluorinated nucleotide analog of the antiviral agent

Cedar Knolls, NJ


vidarabine, 9-b-D-arabinofuranosyladenine (ara-A))


Fluorouracil, 5-FU
Adrucil
ICN Pharmaceuticals, Inc.,


(5-fluoro-2,4(1H,3H)-pyrimidinedione)

Humacao, Puerto Rico


Fulvestrant
Faslodex
IPR Pharmaceuticals,


(7-alpha-[9-(4,4,5,5,5-penta fluoropentylsulphinyl)

Guayama, Puerto Rico


nonyl]estra-1,3,5-(10)-triene-3,17-beta-diol)


Gemcitabine
Gemzar
Eli Lilly


(2′-deoxy-2′,2′-difluorocytidine monohydrochloride (b-


isomer))


Gemtuzumab Ozogamicin
Mylotarg
Wyeth Ayerst


(anti-CD33 hP67.6)


Goserelin acetate
Zoladex
AstraZeneca Pharmaceuticals


(acetate salt of [D-Ser(But)6,Azgly10]LHRH; pyro-Glu-
Implant


His-Trp-Ser-Tyr-D-Ser(But)-Leu-Arg-Pro-Azgly-NH2


acetate [C59H84N18O14•(C2H4O2)x


Hydroxyurea
Hydrea
Bristol-Myers Squibb


Ibritumomab Tiuxetan
Zevalin
Biogen IDEC, Inc.,


(immunoconjugate resulting from a thiourea covalent

Cambridge MA


bond between the monoclonal antibody Ibritumomab and


the linker-chelator tiuxetan [N-[2-


bis(carboxymethyl)amino]-3-(p-isothiocyanatophenyl)-


propyl]-[N-[2-bis(carboxymethyl)amino]-2-(methyl)-


ethyl]glycine)


Idarubicin
Idamycin
Pharmacia & Upjohn


(5,12-Naphthacenedione, 9-acetyl-7-[(3-amino-2,3,6-

Company


trideoxy-(alpha)-L-lyxo-hexopyranosyl)oxy]-7,8,9,10-


tetrahydro-6,9,11-trihydroxyhydrochloride, (7S-cis))


Ifosfamide
IFEX
Bristol-Myers Squibb


(3-(2-chloroethyl)-2-[(2-chloroethyl)amino]tetrahydro-


2H-1,3,2-oxazaphosphorine 2-oxide)


Imatinib Mesilate
Gleevec
Novartis AG, Basel,


(4-[(4-Methyl-1-piperazinyl)methyl]-N-[4-methyl-3-[[4-

Switzerland


(3-pyridinyl)-2-pyrimidinyl]amino]-phenyl]benzamide


methanesulfonate)


Interferon alfa-2a
Roferon-A
Hoffmann-La Roche, Inc.,


(recombinant peptide)

Nutley, NJ


Interferon alfa-2b
Intron A
Schering AG, Berlin,


(recombinant peptide)
(Lyophilized
Germany



Betaseron)


Irinotecan HCl
Camptosar
Pharmacia & Upjohn


((4S)-4,11-diethyl-4-hydroxy-9-[(4-piperi-

Company


dinopiperidino)carbonyloxy]-1H-pyrano[3′,4′: 6,7]


indolizino[1,2-b] quinoline-3,14(4H,12H) dione


hydrochloride trihydrate)


Lenalidomide
Revlimid
Celgene


3-(4-amino-1-oxo 1,3-dihydro-2H-isoindol-2-yl)


piperidine-2,6-dione


Letrozole
Femara
Novartis


(4,4′-(1H-1,2,4-Triazol-1-ylmethylene) dibenzonitrile)


Leucovorin
Wellcovorin,
Immunex, Corp., Seattle, WA


(L-Glutamic acid, N[4[[(2amino-5-formyl1,4,5,6,7,8-
Leucovorin


hexahydro4oxo6-pteridinyl)methyl]amino]benzoyl],


calcium salt (1:1))


Levamisole HCl
Ergamisol
Janssen Research Foundation,


((−)-(S)-2,3,5,6-tetrahydro-6-phenylimidazo [2,1-b]

Titusville, NJ


thiazole monohydrochloride C11H12N2S•HCl)


Lomustine
CeeNU
Bristol-Myers Squibb


(1-(2-chloro-ethyl)-3-cyclohexyl-1-nitrosourea)


Meclorethamine, nitrogen mustard
Mustargen
Merck


(2-chloro-N-(2-chloroethyl)-N-methylethanamine


hydrochloride)


Megestrol acetate
Megace
Bristol-Myers Squibb


17α(acetyloxy)-6-methylpregna-4,6-diene-3,20-dione


Melphalan, L-PAM
Alkeran
GlaxoSmithKline


(4-[bis(2-chloroethyl) amino]-L-phenylalanine)


Mercaptopurine, 6-MP
Purinethol
GlaxoSmithKline


(1,7-dihydro-6H-purine-6-thione monohydrate)


Mesna
Mesnex
Asta Medica


(sodium 2-mercaptoethane sulfonate)


Methotrexate
Methotrexate
Lederle Laboratories


(N-[4-[[(2,4-diamino-6-


pteridinyl)methyl]methylamino]benzoyl]-L-glutamic


acid)


Methoxsalen
Uvadex
Therakos, Inc., Way Exton, Pa


(9-methoxy-7H-furo[3,2-g][1]-benzopyran-7-one)


Mitomycin C
Mutamycin
Bristol-Myers Squibb


mitomycin C
Mitozytrex
SuperGen, Inc., Dublin, CA


Mitotane
Lysodren
Bristol-Myers Squibb


(1,1-dichloro-2-(o-chlorophenyl)-2-(p-chlorophenyl)


ethane)


Mitoxantrone
Novantrone
Immunex Corporation


(1,4-dihydroxy-5,8-bis[[2-[(2-


hydroxyethyl)amino]ethyl]amino]-9,10-anthracenedione


dihydrochloride)


Nandrolone phenpropionate
Durabolin-50
Organon, Inc., West Orange,




NJ


Nofetumomab
Verluma
Boehringer Ingelheim Pharma




KG, Germany


Oprelvekin
Neumega
Genetics Institute, Inc.,


(IL-11)

Alexandria, VA


Oxaliplatin
Eloxatin
Sanofi Synthelabo, Inc., NY, NY


(cis-[(1R,2R)-1,2-cyclohexanediamine-N,N′] [oxalato(2-)-


O,O′] platinum)


Paclitaxel
TAXOL
Bristol-Myers Squibb


(5β,20-Epoxy-1,2a,4,7β,10β,13a-hexahydroxytax-11-


en-9-one 4,10-diacetate 2-benzoate 13-ester with (2R,3S)-


N-benzoyl-3-phenylisoserine)


Pamidronate
Aredia
Novartis


(phosphonic acid (3-amino-1-hydroxypropylidene) bis-,


disodium salt, pentahydrate, (APD))


Pegademase
Adagen
Enzon Pharmaceuticals, Inc.,


((monomethoxypolyethylene glycol succinimidyl) 11-17-
(Pegademase
Bridgewater, NJ


adenosine deaminase)
Bovine)


Pegaspargase
Oncaspar
Enzon


(monomethoxypolyethylene glycol succinimidyl


L-asparaginase)


Pegfilgrastim
Neulasta
Amgen, Inc


(covalent conjugate of recombinant methionyl human G-


CSF (Filgrastim) and monomethoxypolyethylene glycol)


Pentostatin
Nipent
Parke-Davis Pharmaceutical




Co., Rockville, MD


Pipobroman
Vercyte
Abbott Laboratories, Abbott




Park, IL


Plicamycin, Mithramycin
Mithracin
Pfizer, Inc., NY, NY


(antibiotic produced by Streptomyces plicatus)


Porfimer sodium
Photofrin
QLT Phototherapeutics, Inc.,




Vancouver, Canada


Procarbazine
Matulane
Sigma Tau Pharmaceuticals,


(N-isopropyl-μ-(2-methylhydrazino)-p-toluamide

Inc., Gaithersburg, MD


monohydrochloride)


Quinacrine
Atabrine
Abbott Labs


(6-chloro-9-(1-methyl-4-diethyl-amine) butylamino-2-


methoxyacridine)


Rasburicase
Elitek
Sanofi-Synthelabo, Inc.,


(recombinant peptide)


Rituximab
Rituxan
Genentech, Inc., South San


(recombinant anti-CD20 antibody)

Francisco, CA


Sargramostim
Prokine
Immunex Corp


(recombinant peptide)


Streptozocin
Zanosar
Pharmacia & Upjohn


(streptozocin 2-deoxy-2-

Company


[(methylnitrosoamino)carbonyl]amino]-a(and b)-D-


glucopyranose and 220 mg citric acid anhydrous)


Talc
Sclerosol
Bryan, Corp., Woburn, MA


(Mg3Si4O10 (OH)2)


Tamoxifen
Nolvadex
AstraZeneca Pharmaceuticals


((Z)2-[4-(1,2-diphenyl-1-butenyl) phenoxy]-N,N-


dimethylethanamine 2-hydroxy-1,2,3-


propanetricarboxylate (1:1))


Temozolomide
Temodar
Schering


(3,4-dihydro-3-methyl-4-oxoimidazo[5,1-d]-as-tetrazine-


8-carboxamide)


teniposide, VM-26
Vumon
Bristol-Myers Squibb


(4′-demethylepipodophyllotoxin 9-[4,6-0-(R)-2-


thenylidene-(beta)-D-glucopyranoside])


Testolactone
Teslac
Bristol-Myers Squibb


(13-hydroxy-3-oxo-13,17-secoandrosta-1,4-dien-17-oic


acid [dgr]-lactone)


Thioguanine, 6-TG
Thioguanine
GlaxoSmithKline


(2-amino-1,7-dihydro-6 H - purine-6-thione)


Thiotepa
Thioplex
Immunex Corporation


(Aziridine,1,1′,1′′-phosphinothioylidynetris-, or Tris (1-


aziridinyl) phosphine sulfide)


Topotecan HCl
Hycamtin
GlaxoSmithKline


((S)-10-[(dimethylamino) methyl]-4-ethyl-4,9-dihydroxy-


1H-pyrano[3′,4′: 6,7] indolizino [1,2-b] quinoline-3,14-


4H,12H)-dione monohydrochloride)


Toremifene
Fareston
Roberts Pharmaceutical Corp.,


(2-(p-[(Z)-4-chloro-1,2-diphenyl-1-butenyl]-phenoxy)-

Eatontown, NJ


N,N-dimethylethylamine citrate (1:1))


Tositumomab, I 131 Tositumomab
Bexxar
Corixa Corp., Seattle, WA


(recombinant murine immunotherapeutic monoclonal


IgG2a lambda anti-CD20 antibody (I 131 is a


radioimmunotherapeutic antibody))


Trastuzumab
Herceptin
Genentech, Inc


(recombinant monoclonal IgG1 kappa anti-HER2


antibody)


Tretinoin, ATRA
Vesanoid
Roche


(all-trans retinoic acid)


Uracil Mustard
Uracil
Roberts Labs



Mustard



Capsules


Valrubicin, N-trifluoroacetyladriamycin-14-valerate
Valstar
Anthra --> Medeva


((2S-cis)-2-[1,2,3,4,6,11-hexahydro-2,5,12-trihydroxy-7


methoxy-6,11-dioxo-[[4 2,3,6-trideoxy-3-


[(trifluoroacetyl)-amino-α-L-lyxo-hexopyranosyl]oxyl]-2-


naphthacenyl]-2-oxoethyl pentanoate)


Vinblastine, Leurocristine
Velban
Eli Lilly


(C46H56N4O10•H2SO4)


Vincristine
Oncovin
Eli Lilly


(C46H56N4O10•H2SO4)


Vinorelbine
Navelbine
GlaxoSmithKline


(3′,4′-didehydro-4′-deoxy-C′-norvincaleukoblastine [R-


(R*,R*)-2,3-dihydroxybutanedioate (1:2)(salt)])


Zoledronate, Zoledronic acid
Zometa
Novartis


((1-Hydroxy-2-imidazol-1-yl-phosphonoethyl)


phosphonic acid monohydrate)









Anticancer agents further include compounds which have been identified to have anticancer activity but are not currently approved by the U.S. Food and Drug Administration or other counterpart agencies or are undergoing evaluation for new uses. Examples include, but are not limited to, 3-AP, 12-O-tetradecanoylphorbol-13-acetate, 17AAG, 852A, ABI-007, ABR-217620, ABT-751, ADI-PEG 20, AE-941, AG-013736, AGRO100, alanosine, AMG 706, antibody G250, antineoplastons, AP23573, apaziquone, AP C8015, atiprimod, ATN-161, atrasenten, azacitidine, BB-10901, BCX-1777, bevacizumab, BG00001, bicalutamide, BMS 247550, bortezomib, bryostatin-1, buserelin, calcitriol, CCl-779, CDB-2914, cefixime, cetuximab, CG0070, cilengitide, clofarabine, combretastatin A4 phosphate, CP-675,206, CP-724,714, CpG 7909, curcumin, decitabine, DENSPM, doxercalciferol, E7070, E7389, ecteinascidin 743, efaproxiral, eflornithine, EKB-569, enzastaurin, erlotinib, exisulind, fenretinide, flavopiridol, fludarabine, flutamide, fotemustine, FR901228, Gl7DT, galiximab, gefitinib, genistein, glufosfamide, GTI-2040, histrelin, HKI-272, homoharringtonine, HSPPC-96, hu14.18-interleukin-2 fusion protein, HuMax-CD4, iloprost, imiquimod, infliximab, interleukin-12, IPI-504, irofulven, ixabepilone, lapatinib, lestaurtinib, leuprolide, LMB-9 immunotoxin, lonafarnib, luniliximab, mafosfamide, MB07133, MDX-010, MLN2704, monoclonal antibody 3F8, monoclonal antibody J591, motexafin, MS-275, MVA-MUC1-IL2, nilutamide, nitrocamptothecin, nolatrexed dihydrochloride, nolvadex, NS-9, O6-benzylguanine, oblimersen sodium, ONYX-015, oregovomab, OSI-774, panitumumab, paraplatin, PD-0325901, pemetrexed, PHY906, pioglitazone, pirfenidone, pixantrone, PS-341, PSC 833, PXD101, pyrazoloacridine, R115777, RAD001, ranpirnase, rebeccamycin analogue, rhuAngiostatin protein, rhuMab 2C4, rosiglitazone, rubitecan, S-1, S-8184, satraplatin, SB-, 15992, SGN-0010, SGN-40, sorafenib, SR31747A, ST1571, SU011248, suberoylanilide hydroxamic acid, suramin, talabostat, talampanel, tariquidar, temsirolimus, TGFa-PE38 immunotoxin, thalidomide, thymalfasin, tipifarnib, tirapazamine, TLK286, trabectedin, trimetrexate glucuronate, TroVax, UCN-1, valproic acid, vinflunine, VNP40101M, volociximab, vorinostat, VX-680, ZD1839, ZD6474, zileuton, and zosuquidar trihydrochloride.
In one embodiment, the anticancer agent is selected from the group consisting of taxotere, gemcitabine, lapatinib)(Tykerb®) and etoposide.
For a more detailed description of anticancer agents and other therapeutic agents, those skilled in the art are referred to any number of instructive manuals including, but not limited to, the Physician's Desk Reference and to Goodman and Gilman's “Pharmaceutical Basis of Therapeutics” tenth edition, Eds. Hardman et al., 2002.
The present invention provides methods for administering a compound of Formulae I-XIa with radiation therapy. The invention is not limited by the types, amounts, or delivery and administration systems used to deliver the therapeutic dose of radiation to an animal. For example, the animal may receive photon radiotherapy, particle beam radiation therapy, other types of radiotherapies, and combinations thereof. In some embodiments, the radiation is delivered to the animal using a linear accelerator. In still other embodiments, the radiation is delivered using a gamma knife.
The source of radiation can be external or internal to the animal. External radiation therapy is most common and involves directing a beam of high-energy radiation to a tumor site through the skin using, for instance, a linear accelerator. While the beam of radiation is localized to the tumor site, it is nearly impossible to avoid exposure of normal, healthy tissue. However, external radiation is usually well tolerated by animals. Internal radiation therapy involves implanting a radiation-emitting source, such as beads, wires, pellets, capsules, particles, and the like, inside the body at or near the tumor site including the use of delivery systems that specifically target cancer cells (e.g., using particles attached to cancer cell binding ligands). Such implants can be removed following treatment, or left in the body inactive. Types of internal radiation therapy include, but are not limited to, brachytherapy, interstitial irradiation, intracavity irradiation, radioimmunotherapy, and the like.
The animal may optionally receive radiosensitizers (e.g., metronidazole, misonidazole, intra-arterial Budr, intravenous iododeoxyuridine (IudR), nitroimidazole, 5-substituted-4-nitroimidazoles, 2H-isoindolediones, [[(2-bromoethyl)-amino]methyl]-nitro-1H-imidazole-1-ethanol, nitroaniline derivatives, DNA-affinic hypoxia selective cytotoxins, halogenated DNA ligand, 1,2,4 benzotriazine oxides, 2-nitroimidazole derivatives, fluorine-containing nitroazole derivatives, benzamide, nicotinamide, acridine-intercalator, 5-thiotretrazole derivative, 3-nitro-1,2,4-triazole, 4,5-dinitroimidazole derivative, hydroxylated texaphrins, cisplatin, mitomycin, tiripazamine, nitrosourea, mercaptopurine, methotrexate, fluorouracil, bleomycin, vincristine, carboplatin, epirubicin, doxorubicin, cyclophosphamide, vindesine, etoposide, paclitaxel, heat (hyperthermia), and the like), radioprotectors (e.g., cysteamine, aminoalkyl dihydrogen phosphorothioates, amifostine (WR 2721), IL-1, IL-6, and the like). Radiosensitizers enhance the killing of tumor cells. Radioprotectors protect healthy tissue from the harmful effects of radiation.
Any type of radiation can be administered to an animal, so long as the dose of radiation is tolerated by the patient without unacceptable negative side-effects. Suitable types of radiotherapy include, for example, ionizing (electromagnetic) radiotherapy (e.g., X-rays or gamma rays) or particle beam radiation therapy (e.g., high linear energy radiation). Ionizing radiation is defined as radiation comprising particles or photons that have sufficient energy to produce ionization, i.e., gain or loss of electrons (as described in, for example, U.S. Pat. No. 5,770,581 incorporated herein by reference in its entirety). The effects of radiation can be at least partially controlled by the clinician. The dose of radiation is preferably fractionated for maximal target cell exposure and reduced toxicity.
The total dose of radiation administered to an animal preferably is about 0.01 Gray (Gy) to about 100 Gy. More preferably, about 10 Gy to about 65 Gy (e.g., about 15 Gy, 20 Gy, 25 Gy, 30 Gy, 35 Gy, 40 Gy, 45 Gy, 50 Gy, 55 Gy, or 60 Gy) are administered over the course of treatment. While in some embodiments a complete dose of radiation can be administered over the course of one day, the total dose is ideally fractionated and administered over several days. Desirably, radiotherapy is administered over the course of at least about 3 days, e.g., at least 5, 7, 10, 14, 17, 21, 25, 28, 32, 35, 38, 42, 46, 52, or 56 days (about 1-8 weeks). Accordingly, a daily dose of radiation will comprise approximately 1-5 Gy (e.g., about 1 Gy, 1.5 Gy, 1.8 Gy, 2 Gy, 2.5 Gy, 2.8 Gy, 3 Gy, 3.2 Gy, 3.5 Gy, 3.8 Gy, 4 Gy, 4.2 Gy, or 4.5 Gy), preferably 1-2 Gy (e.g., 1.5-2 Gy). The daily dose of radiation should be sufficient to induce destruction of the targeted cells. If stretched over a period, radiation preferably is not administered every day, thereby allowing the animal to rest and the effects of the therapy to be realized. For example, radiation desirably is administered on 5 consecutive days, and not administered on 2 days, for each week of treatment, thereby allowing 2 days of rest per week. However, radiation can be administered 1 day/week, 2 days/week, 3 days/week, 4 days/week, 5 days/week, 6 days/week, or all 7 days/week, depending on the animal's responsiveness and any potential side effects. Radiation therapy can be initiated at any time in the therapeutic period. Preferably, radiation is initiated in week 1 or week 2, and is administered for the remaining duration of the therapeutic period. For example, radiation is administered in weeks 1-6 or in weeks 2-6 of a therapeutic period comprising 6 weeks for treating, for instance, a solid tumor. Alternatively, radiation is administered in weeks 1-5 or weeks 2-5 of a therapeutic period comprising 5 weeks. These exemplary radiotherapy administration schedules are not intended, however, to limit the present invention.
Antimicrobial therapeutic agents may also be used as therapeutic agents in the present invention. Any agent that can kill, inhibit, or otherwise attenuate the function of microbial organisms may be used, as well as any agent contemplated to have such activities. Antimicrobial agents include, but are not limited to, natural and synthetic antibiotics, antibodies, inhibitory proteins (e.g., defensins), antisense nucleic acids, membrane disruptive agents and the like, used alone or in combination. Indeed, any type of antibiotic may be used including, but not limited to, antibacterial agents, antiviral agents, antifungal agents, and the like.
In some embodiments of the present invention, a compound of Formulae I-XIa and one or more therapeutic agents or anticancer agents are administered to an animal under one or more of the following conditions: at different periodicities, at different durations, at different concentrations, by different administration routes, in a single composition, in separate compositions, etc. In some embodiments, the compound is administered prior to the therapeutic or anticancer agent, e.g., 0.5, 1, 2, 3, 4, 5, 10, 12, or 18 hours, 1, 2, 3, 4, 5, or 6 days, 1, 2, 3, or 4 weeks prior to the administration of the therapeutic or anticancer agent. In some embodiments, the compound is administered after the therapeutic or anticancer agent, e.g., 0.5, 1, 2, 3, 4, 5, 10, 12, or 18 hours, 1, 2, 3, 4, 5, or 6 days, 1, 2, 3, or 4 weeks after the administration of the anticancer agent. In some embodiments, the compound and the therapeutic or anticancer agent are administered concurrently but on different schedules, e.g., the compound is administered daily while the therapeutic or anticancer agent is administered once a week, once every two weeks, once every three weeks, or once every four weeks. In other embodiments, the compound is administered once a week while the therapeutic or anticancer agent is administered daily, once a week, once every two weeks, once every three weeks, or once every four weeks.
Compositions within the scope of this invention include all compositions wherein the compounds of the present invention are contained in an amount which is effective to achieve its intended purpose. While individual needs vary, determination of optimal ranges of effective amounts of each component is within the skill of the art. Typically, the compounds may be administered to mammals, e.g. humans, orally at a dose of 0.0025 to 50 mg/kg, or an equivalent amount of the pharmaceutically acceptable salt thereof, per day of the body weight of the mammal being treated for disorders responsive to induction of apoptosis. For example, about 0.01 to about 25 mg/kg is orally administered to treat, ameliorate, or prevent such disorders. For intramuscular injection, the dose is generally about one-half of the oral dose. For example, a suitable intramuscular dose would be about 0.0025 to about 25 mg/kg, e.g., from about 0.01 to about 5 mg/kg.
The unit oral dose may comprise from about 0.01 to about 1000 mg, e.g., about 0.1 to about 100 mg of the compound. The unit dose may be administered one or more times daily as one or more tablets or capsules each containing from about 0.1 to about 10, conveniently about 0.25 to 50 mg of the compound or its solvates.
In a topical formulation, the compound may be present at a concentration of about 0.01 to 100 mg per gram of carrier. In one embodiment, the compound is present at a concentration of about 0.07-1.0 mg/ml, e.g., about 0.1-0.5 mg/ml, e.g., about 0.4 mg/ml.
In addition to administering the compound as a raw chemical, the compounds of the invention may be administered as part of a pharmaceutical preparation containing suitable pharmaceutically acceptable carriers comprising excipients and auxiliaries which facilitate processing of the compounds into preparations which can be used pharmaceutically. Preferably, the preparations, particularly those preparations which can be administered orally or topically and which can be used for the preferred type of administration, such as tablets, dragees, slow release lozenges and capsules, mouth rinses and mouth washes, gels, liquid suspensions, hair rinses, hair gels, shampoos and also preparations which can be administered rectally, such as suppositories, as well as suitable solutions for administration by intravenous infusion, injection, topically or orally, contain from about 0.01 to 99 percent, e.g., from about 0.25 to 75 percent of active compound(s), together with the excipient.
The pharmaceutical compositions of the invention may be administered to any animal which may experience the beneficial effects of the compounds of the invention. Foremost among such animals are mammals, e.g., humans, although the invention is not intended to be so limited. Other animals include veterinary animals (cows, sheep, pigs, horses, dogs, cats and the like).
The compounds and pharmaceutical compositions thereof may be administered by any means that achieve their intended purpose. For example, administration may be by parenteral, subcutaneous, intravenous, intramuscular, intraperitoneal, transdermal, buccal, intrathecal, intracranial, intranasal or topical routes. Alternatively, or concurrently, administration may be by the oral route. The dosage administered will be dependent upon the age, health, and weight of the recipient, kind of concurrent treatment, if any, frequency of treatment, and the nature of the effect desired.
The pharmaceutical preparations of the present invention are manufactured in a manner which is itself known, for example, by means of conventional mixing, granulating, dragee-making, dissolving, or lyophilizing processes. Thus, pharmaceutical preparations for oral use can be obtained by combining the active compounds with solid excipients, optionally grinding the resulting mixture and processing the mixture of granules, after adding suitable auxiliaries, if desired or necessary, to obtain tablets or dragee cores.
Suitable excipients are, in particular, fillers such as saccharides, for example lactose or sucrose, mannitol or sorbitol, cellulose preparations and/or calcium phosphates, for example tricalcium phosphate or calcium hydrogen phosphate, as well as binders such as starch paste, using, for example, maize starch, wheat starch, rice starch, potato starch, gelatin, tragacanth, methyl cellulose, hydroxypropylmethylcellulose, sodium carboxymethylcellulose, and/or polyvinyl pyrrolidone. If desired, disintegrating agents may be added such as the above-mentioned starches and also carboxymethyl-starch, cross-linked polyvinyl pyrrolidone, agar, or alginic acid or a salt thereof, such as sodium alginate. Auxiliaries are, above all, flow-regulating agents and lubricants, for example, silica, talc, stearic acid or salts thereof, such as magnesium stearate or calcium stearate, and/or polyethylene glycol. Dragee cores are provided with suitable coatings which, if desired, are resistant to gastric juices. For this purpose, concentrated saccharide solutions may be used, which may optionally contain gum arabic, talc, polyvinyl pyrrolidone, polyethylene glycol and/or titanium dioxide, lacquer solutions and suitable organic solvents or solvent mixtures. In order to produce coatings resistant to gastric juices, solutions of suitable cellulose preparations such as acetylcellulose phthalate or hydroxypropylmethyl-cellulose phthalate, are used. Dye stuffs or pigments may be added to the tablets or dragee coatings, for example, for identification or in order to characterize combinations of active compound doses.
Other pharmaceutical preparations which can be used orally include push-fit capsules made of gelatin, as well as soft, sealed capsules made of gelatin and a plasticizer such as glycerol or sorbitol. The push-fit capsules can contain the active compounds in the form of granules which may be mixed with fillers such as lactose, binders such as starches, and/or lubricants such as talc or magnesium stearate and, optionally, stabilizers. In soft capsules, the active compounds are preferably dissolved or suspended in suitable liquids, such as fatty oils, or liquid paraffin. In addition, stabilizers may be added.
Possible pharmaceutical preparations which can be used rectally include, for example, suppositories, which consist of a combination of one or more of the active compounds with a suppository base. Suitable suppository bases are, for example, natural or synthetic triglycerides, or paraffin hydrocarbons. In addition, it is also possible to use gelatin rectal capsules which consist of a combination of the active compounds with a base. Possible base materials include, for example, liquid triglycerides, polyethylene glycols, or paraffin hydrocarbons.
Suitable formulations for parenteral administration include aqueous solutions of the active compounds in water-soluble form, for example, water-soluble salts and alkaline solutions. In addition, suspensions of the active compounds as appropriate oily injection suspensions may be administered. Suitable lipophilic solvents or vehicles include fatty oils, for example, sesame oil, or synthetic fatty acid esters, for example, ethyl oleate or triglycerides or polyethylene glycol-400. Aqueous injection suspensions may contain substances which increase the viscosity of the suspension include, for example, sodium carboxymethyl cellulose, sorbitol, and/or dextran. Optionally, the suspension may also contain stabilizers.
The topical compositions of this invention are formulated preferably as oils, creams, lotions, ointments and the like by choice of appropriate carriers. Suitable carriers include vegetable or mineral oils, white petrolatum (white soft paraffin), branched chain fats or oils, animal fats and high molecular weight alcohol (greater than C12). The preferred carriers are those in which the active ingredient is soluble. Emulsifiers, stabilizers, humectants and antioxidants may also be included as well as agents imparting color or fragrance, if desired. Additionally, transdermal penetration enhancers can be employed in these topical formulations. Examples of such enhancers can be found in U.S. Pat. Nos. 3,989,816 and 4,444,762.
Creams are preferably formulated from a mixture of mineral oil, self-emulsifying beeswax and water in which mixture the active ingredient, dissolved in a small amount of an oil such as almond oil, is admixed. A typical example of such a cream is one which includes about 40 parts water, about 20 parts beeswax, about 40 parts mineral oil and about 1 part almond oil.
Ointments may be formulated by mixing a solution of the active ingredient in a vegetable oil such as almond oil with warm soft paraffin and allowing the mixture to cool. A typical example of such an ointment is one which includes about 30% almond oil and about 70% white soft paraffin by weight.
Lotions may be conveniently prepared by dissolving the active ingredient, in a suitable high molecular weight alcohol such as propylene glycol or polyethylene glycol.
In certain aspects, the present invention is drawn to the following particular embodiments:
I. A compound having Formula I:




    
    
        wherein:
    
    


A1 and A2 are independently selected from the group consisting of hydrogen and optionally substituted alkyl, wherein A2 is absent when V is O;
V is selected from the group consisting of N, CH and O;
W is selected from the group consisting of CH and N;
X is selected from the group consisting of hydrogen, optionally substituted alkyl and aralkyl;
Y is selected from the group consisting of CON(R1), N(R1)CO, C(O)O, OC(O), (CH2)1-3, wherein one or more CH2 groups can be replaced by O, S, or NR1, optionally substituted aryl and optionally substituted heteroaryl;
Z is (CR2aR2b);
D is (CR3aR3b)n—U—(CR4aR4b)m;
U is selected from the group consisting of CR5aR5b and NR6;
J is (CR7aR7b)p-L-(CR8aR8b)q; 
T is optionally substituted heteroaryl;
n, m, p and q are independently selected from the group consisting of 0-5;
r is 0-3;
R1 is selected from the group consisting of hydrogen, optionally substituted alkyl, aralkyl, optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heteroaryl and optionally substituted heterocyclo;
each R2a, R2b, R3a, R3b, R4a, R4b, R5a, R5b, R7a, R7b, R8a, R8b are independently selected from the group consisting of hydrogen, optionally substituted alkyl, aralkyl, optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heteroaryl and optionally substituted heterocyclo;
R6 is selected from the group consisting of hydrogen, optionally substituted alkyl, aralkyl, optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heteroaryl and optionally substituted heterocyclo and COR9;
L is selected from the group consisting of O, S, NR1, NCOR9, CR7aR7b, C═O, C═S and C═NR1; and
R9 is selected from the group consisting of hydrogen, optionally substituted alkyl, aralkyl, optionally substituted cycloalkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted aryl, optionally substituted heteroaryl and optionally substituted heterocyclo;
or pharmaceutically acceptable salt or prodrug thereof.
II. The compound of I, wherein n is l, m is 1 or 2, p is 0, L is CR7aR7b, q is 1, and R3a, R3b, R4a, R4b, R7a, R7b, R8a, R8b are hydrogen.
III. The compound of II, wherein Y is CON(H), W is CH, r is 0 and V is N.
IV. The compound of I having formula II:




V. The compound of IV, wherein n is l, m is 1 or 2, p is 0, L is CH2, q is 1, and R3a, R3b, R4a, R4b, R7a, R7b, R8a, R8b are hydrogen.
VI. The compound of V, Y is CON(H), W is CH, r is 0 and V is N.
VII. The compound of VI having Formula V:




VIII. The compound of VII, wherein A1 is optionally substituted alkyl and A2 is hydrogen.
IX. The compound of VII, wherein X is optionally substituted alkyl.
X. The compound of VII, wherein U is CH2.
XI. The compound of VII, wherein U is NR6.
XII. The compound of XI, wherein R6 is COR9.
XIII. The compound of XII, wherein R9 is selected from the group consisting of optionally substituted alkyl and aralkyl.
XIV. The compound of VII, wherein m is 2.
XV. The compound of VII, wherein m is 1.
XVI. The compound of VII, wherein A1 is optionally substituted alkyl, A2 is hydrogen, X is optionally substituted alkyl, U is NR6, R6 is COR9, R9 is selected from the group consisting of optionally substituted alkyl and aralkyl, and m is 1.
XVII. The compound of I-XV, wherein T is selected from the group consisting of




wherein:
Q is selected from the group consisting of O, S and NR12;
R12 is selected from the group consisting of hydrogen, optionally substituted alkyl, aralkyl, optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heteroaryl and optionally substituted heterocyclo;
R10a, R10b, R11a, R11b, R11c and R11d are independently selected from the group consisting of hydrogen, optionally substituted alkyl, haloalkyl, aralkyl, optionally substituted cycloalkyl, optionally substituted cycloalkenyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocyclo, halo, nitro, cyano, hydroxy, amino, alkoxy, aryloxy, arylalkyloxy, alkylthio, carboxamido and sulfonamido;
Z1, Z2, and Z3 are independently selected from the group consisting of CR11e and N, wherein at least one of Z1, Z2, and Z3 is CR11e, and at least one of Z1, Z2, and Z3 is N; and
R11e is selected from the group consisting of hydrogen, optionally substituted alkyl, haloalkyl, aralkyl, optionally substituted cycloalkyl, optionally substituted cycloalkenyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocyclo, halo, nitro, cyano, hydroxy, amino, alkoxy, aryloxy, arylalkyloxy, alkylthio, carboxamido and sulfonamido.
XVIII. The compound of XVI, wherein T is selected from the group consisting of




wherein:
Q is selected from the group consisting of O, S and NR12;
R12 is selected from the group consisting of hydrogen, optionally substituted alkyl, aralkyl, optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heteroaryl and optionally substituted heterocyclo; and
R10a, R10b, R11a, R11b, R11c and R11d are independently selected from the group consisting of hydrogen, optionally substituted alkyl, haloalkyl, aralkyl, optionally substituted cycloalkyl, optionally substituted cycloalkenyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocyclo, halo, nitro, cyano, hydroxy, amino, alkoxy, aryloxy, arylalkyloxy, alkylthio, carboxamido and sulfonamido.
XIX. The compound of XVIII wherein T is




XX. The compound of XIX, wherein R10a is optionally substituted aryl and R10b is hydrogen.
XXI. The compound of XVIII, wherein T is




XXII. The compound of XXI, wherein R11a is selected from the group consisting of optionally substituted aryl, aralkyl, and optionally substituted alkyl; and R11b, R11c and R11d are each hydrogen.
XXIII. The compound of XXI, wherein R11d is selected from the group consisting of optionally substituted aryl, aralkyl, and optionally substituted alkyl; R11a, R11b and R11c are each hydrogen.
XXIV. The compound of XXII having Formula XIa:




wherein R9 is optionally substituted alkyl or aralkyl, and R11a is optionally substituted aryl.

XXV. The compound of I, selected from the group consisting of:










or a pharmaceutically acceptable salt or prodrug thereof.

XXVI. A pharmaceutical composition comprising the compound of any one of I-XXV and a pharmaceutically acceptable carrier.
XXVII. A method of inducing apoptosis in a cell comprising contacting the cell with the compound of any one of I-XXV.)
XXVIII. A method of rendering a cell sensitive to an inducer of apoptosis comprising contacting the cell with the compound of any one of I-XXV.
XXIX. The method of XXVIII, further comprising contacting the cell with an inducer of apoptosis.
XXX. The method of XXIX, wherein said inducer of apoptosis is a chemotherapeutic agent.
XXXI. The method of XXIX, wherein said inducer of apoptosis is radiation.
XXXII. The method of XXIX, wherein said inducer of apoptosis is a tumor necrosis factor (TNF), a TNF-related ligand, or an agonist of TRAIL-R1 or TRAIL-R2.
XXXIII. The method of XXXII, wherein said TNF-related ligand is selected from the group consisting of a TRAMP ligand, a Fas/CD95 ligand, a TNFR-1 ligand, and TRAIL.
XXXIV. The method of XXXIII, wherein said TNF-related ligand is TRAIL.
XXXV. The method of XXXIV, wherein said agonist of TRIAL-R1 or TRAIL-R2 is an antibody.)
XXXVI. A method of treating, ameliorating, or preventing a disorder responsive to the induction of apoptosis in an animal, comprising administering to said animal a therapeutically effective amount of the compound of any one of 1-XXV.)
XXXVII. The method of XXXVI, further comprising administering an inducer of apoptosis.
XXXVIII. The method of XXXVII, wherein said inducer of apoptosis is a chemotherapeutic agent.
XXXIX. The method of XXXVIII, wherein said inducer of apoptosis is radiation.
XL. The method of XXXVII, wherein said inducer of apoptosis is a TNF, a TNF-related ligand, or an agonist of TRAIL-R1 or TRAIL-R2.
XLI. The method of XL, wherein said TNF-related ligand is selected from the group consisting of a TRAMP ligand, a Fas/CD95 ligand, a TNFR-1 ligand, and TRAIL.
XLII. The method of XLI, wherein said TNF-related ligand is TRAIL.
XLIII. The method of XLII, wherein said agonist of TRAIL-R1 or TRAIL-R2 is an antibody.
XLIV. The method of XXXVI, wherein said disorder responsive to the induction of apoptosis is a hyperproliferative disease.
XLV. The method of XLIV, wherein said hyperproliferative disease is cancer.
XLVI. The method of XXXVI, wherein said compound of I-XXV is administered prior to said inducer of apoptosis.
XLVII. The method of XXXVI, wherein said compound of I-XXV is administered after said inducer of apoptosis.
XLVIII. The method of XXXVI, wherein said compound of I-XXV is administered concurrently with said inducer of apoptosis.
XLIX. A method of treating, ameliorating, or preventing a hyperproliferative disease in an animal, comprising administering to said animal a therapeutically effective amount of the compound of any one of I-XXV.
L. The method of XLIX, further comprising administering an anticancer agent.
LI. The method of L, wherein said anticancer agent is an inducer of apoptosis.
LII. The method of LI, wherein said inducer of apoptosis is a chemotherapeutic agent.
LIII. The method of LII, wherein said inducer of apoptosis is radiation.
LIV. The method of LI, wherein said inducer of apoptosis is a TNF, a TNF-related ligand, or an agonist of TRAIL-R1 or TRAIL-R2.
LV. The method of LIV, wherein said TNF-related ligand is selected from the group consisting of a TRAMP ligand, a Fas/CD95 ligand, a TNFR-1 ligand, and TRAIL.
LVI. The method of LIV, wherein said TNF-related ligand is TRAIL.
LVII. The method of LIV, wherein said agonist of TRAIL-R1 or TRAIL-R2 is an antibody.
LVIII. The method of XLIX, wherein said hyperproliferative disease is cancer.
LIX. The method of L, wherein said compound of I-XXV is administered prior to said anticancer agent.
LX. The method of L, wherein said compound of I-XXV is administered after said anticancer agent.
LXI. The method of L, wherein said compound of I-XXV is administered concurrently with said anticancer agent.
LXII. The method of L, wherein said anticancer agent is selected from the group consisting of taxotere, lapatinib and gemcitabine.
LXIII. A method of preventing or inhibiting angiogenesis in an animal in need thereof, comprising administering to said animal a therapeutically effective amount of the compound of any one of I-XXV.
LXIV. The method of LXIII wherein said animal has a disease or disorder selected from the group consisting of macular degeneration, rheumatoid arthritis, psoriasis, diabetic retinopathy, retinopathy of prematurity, corneal graft rejection, neovascular glaucoma, retrolental fibroplasia, rubeosis, Osler-Webber Syndrome, myocardial angiogenesis, plaque neovascularization, telangiectasia, hemophiliac joints, angiofibroma, wound granulation, intestinal adhesions, atherosclerosis, scleroderma and hypertrophic scars.
LXV. A kit comprising the compound of any one of I-XXV and instructions for administering said compound to an animal.
LXVI. The kit of LXV, further comprising an anticancer agent.
LXVII. The kit of LXVI, wherein said anticancer agent is an inducer of apoptosis.
LXVIII. The kit of LXVII wherein said inducer of apoptosis is a chemotherapeutic agent.
LXIX. The kit of LXVII, wherein said inducer of apoptosis is a TNF, a TNF-related ligand, or an agonist of TRAIL-R1 or TRAIL-R2.
LXX. The kit of LXIX, wherein the TNF-related ligand is selected from the group consisting of a TRAMP ligand, a Fas/CD95 ligand, a TNFR-1 ligand, and TRAIL.
LXXI. The kit of LXX, wherein said TNF-related ligand is TRAIL.
LXXII. The kit of LXIX, wherein said agonist of TRAIL-R1 or TRAIL-R2 is an antibody.
LXXIII. The kit of LXV, wherein said instructions are for administering said compound to an animal having a hyperproliferative disease.
LXXIV. The kit of LXXIII, wherein said hyperproliferative disease is cancer.
LXXV. A process for preparing a compound of Formula XII




wherein
R13 is selected from the group consisting of N(H)P1 and




P1 is an amine protecting group;
A1 and A2 are independently selected from the group consisting of hydrogen and optionally substituted alkyl, wherein A2 is absent when V is O;
V is selected from the group consisting of N, CH and O;
W is selected from the group consisting of CH and N;
X is selected from the group consisting of hydrogen, optionally substituted alkyl and aralkyl;
Y is selected from the group consisting of CON(R1), N(R1)CO, C(O)O, OC(O), (CH2)1-3, wherein one or more CH2 groups can be replaced by O, S, or NR1, optionally substituted aryl and optionally substituted heteroaryl;
Z is (CR2aR2b)r; 
U is selected from the group consisting of CR5aR5b and NR6;
m is 1 or 2;
r is 0-3;
R1 is selected from the group consisting of hydrogen, optionally substituted alkyl, aralkyl, optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heteroaryl and optionally substituted heterocyclo;
each R2a, R2b, R5a and R5b are independently selected from the group consisting of hydrogen, optionally substituted alkyl, aralkyl, optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heteroaryl and optionally substituted heterocyclo;
R6 is selected from the group consisting of hydrogen, optionally substituted alkyl, aralkyl, optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heteroaryl and optionally substituted heterocyclo and COR9;
R9 is selected from the group consisting of hydrogen, optionally substituted alkyl, aralkyl, optionally substituted cycloalkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted aryl, optionally substituted heteroaryl and optionally substituted heterocyclo; and
R10a and R10b are independently selected from the group consisting of hydrogen, optionally substituted alkyl, haloalkyl, aralkyl, optionally substituted cycloalkyl, optionally substituted cycloalkenyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted aryl, optionally substituted heteroaryl and optionally substituted heterocyclo, comprising:
a) condensing a compound of Formula XIII




with ammonia to give a compound of Formula XIV




b) converting a compound of Formula XIV to a compound of Formula XV




c) condensing a compound of Formula XV with a compound of Formula XVI,




wherein L2 is a leaving group, to give a compound of Formula XVII,




and b) cyclizing a compound of Formula XVII, to give a compound of Formula XII.
LXXVI. The process of LXXV, wherein L2 is selected from the group consisting of Cl and Br.
LXXVII. The process of LXXVI, wherein L2 is Br.
LXXVIII. The process of LXXVI, wherein R10a is optionally substituted aryl.
LXXIX. The process of LXXV, wherein m is 1.
LXXX. The process of LXXV, wherein R13 is —N(H)P1.
LXXXI. The process of LXXX, wherein P1 is selected from the group consisting of t-butoxycarbonyl and benzyloxycarbonyl.
LXXXII. The process of LXXV, wherein R113 is




LXXXIII. The process of LXXXII, wherein Y is CON(H), W is CH, r is 0 and V is N.
LXXXIV. The process of LXXV, wherein U is NR6 and R6 is COR9.
LXXXV. A process for the preparing a compound of Formula XVIII




wherein
R13 is selected from the group consisting of N(H)P1 and




P1 is an amine protecting group;
A1 and A2 are independently selected from the group consisting of hydrogen and optionally substituted alkyl, wherein A2 is absent when V is O;
V is selected from the group consisting of N, CH and O;
W is selected from the group consisting of CH and N;
X is selected from the group consisting of hydrogen, optionally substituted alkyl and aralkyl;
Y is selected from the group consisting of CON(R1), N(R1)CO, C(O)O, OC(O), (CH2)1-3, wherein one or more CH2 groups can be replaced by O, S, or NR1, optionally substituted aryl and optionally substituted heteroaryl;
Z is (CR2aR2b)r; 
U is selected from the group consisting of CR5aR5b and NR6;
m is 1 or 2;
r is 0-3;
R1 is selected from the group consisting of hydrogen, optionally substituted alkyl, aralkyl, optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heteroaryl and optionally substituted heterocyclo;
each R2a, R2b, R5a and R5b are individually selected from the group consisting of hydrogen, optionally substituted alkyl, aralkyl, optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heteroaryl and optionally substituted heterocyclo;
R6 is selected from the group consisting of hydrogen, optionally substituted alkyl, aralkyl, optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heteroaryl and optionally substituted heterocyclo and COR9;
R9 is selected from the group consisting of hydrogen, optionally substituted alkyl, aralkyl, optionally substituted cycloalkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted aryl, optionally substituted heteroaryl and optionally substituted heterocyclo;
R11a, R11b, R11c and R11d are independently selected from the group consisting of hydrogen, optionally substituted alkyl, haloalkyl, aralkyl, optionally substituted cycloalkyl, optionally substituted cycloalkenyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocyclo, halo, nitro, cyano, hydroxy, amino, alkoxy, aryloxy, arylalkyloxy, alkylthio, carboxamido and sulfonamido, and
R12 is selected from the group consisting of hydrogen, optionally substituted alkyl, aralkyl, optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heteroaryl and optionally substituted heterocyclo, comprising:
a) condensing a compound of Formula XIII




with a compound of Formula XIX




to give a compound of Formula XX




and b) cyclizing a compound of Formula XX, to give a compound of Formula XVIII.
LXXXVI. The process of LXXXV, wherein R12 is hydrogen.
LXXXVII. The process of LXXXV, wherein R11a is optionally substituted phenyl.
LXXXVIII. The process of LXXXV, wherein m is 1.
LXXXIX. The process of LXXXV, wherein R13 is —N(H)P1.
XC. The process of LXXXIX, wherein P1 is selected from the group consisting of t-butoxycarbonyl and benzyloxycarbonyl.
XCI. The process of LXXXV, wherein R13 is




XCII. The process of XCI, wherein Y is CON(H), W is CH, r is 0 and V is N.
XCIII. The process of LXXXV, wherein U is NR6 and R6 is COR9.
XCIV. A process for preparing a compound of Formula XXI




wherein:
A1 and A2 are independently selected from the group consisting of hydrogen and optionally substituted alkyl, wherein A2 is absent when V is O;
V is selected from the group consisting of N, CH and O;
W is selected from the group consisting of CH and N;
X is selected from the group consisting of hydrogen, optionally substituted alkyl and aralkyl;
Z is (CR2aR2b)r; 
U is selected from the group consisting of CR5aR5b and NR6;
m is 1 or 2;
r is 0-3;
each R2a, R2b, R5a and R5b are independently selected from the group consisting of hydrogen, optionally substituted alkyl, aralkyl, optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heteroaryl and optionally substituted heterocyclo;
R6 is selected from the group consisting of hydrogen, optionally substituted alkyl, aralkyl, optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heteroaryl and optionally substituted heterocyclo and COR9;
R9 is selected from the group consisting of hydrogen, optionally substituted alkyl, aralkyl, optionally substituted cycloalkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted aryl, optionally substituted heteroaryl and optionally substituted heterocyclo; and
T is optionally substituted heteroaryl, comprising:
reacting a compound of Formula XXIII




with a compound of Formula XXIV




wherein L1 is a leaving group, to give a compound of Formula XXI.
XCV. The process of XCIV, wherein W is CH and V is N.
XCVI. The process of XCIV wherein T is selected from the group consisting of




wherein:
Q is selected from the group consisting of O, S and NR12;
R12 is selected from the group consisting of hydrogen, optionally substituted alkyl, aralkyl, optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heteroaryl and optionally substituted heterocyclo; and
R10a, R10b, R11a, R11b, R11c and R11d are independently selected from the group consisting of hydrogen, optionally substituted alkyl, haloalkyl, aralkyl, optionally substituted cycloalkyl, optionally substituted cycloalkenyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocyclo, halo, nitro, cyano, hydroxy, amino, alkoxy, aryloxy, arylalkyloxy, alkylthio, carboxamido and sulfonamido.
XCVII. The process of XCIV, wherein U is CH2.
XCVIII. The process of XCIV, wherein U is NCOR9.
XCIX. The process of XCVIII, wherein R9 is optionally substituted alkyl.
C. The process of XCIV, wherein L1 is selected from the group consisting of —Cl and —OH.
CI. The process of C, wherein L1 is —OH and the condensation is carried out in the presence of an activating agent.
CII. The process of XCIV, wherein said compound of Formula XXIII is prepared by removing P1 from a compound of Formula XXII




wherein P1 is an amine protecting group.

CIII. The process of CII, wherein P1 is selected from the group consisting of t-butoxycarbonyl and benzyloxycarbonyl.
CIV. A compound having Formula XXII:




wherein:
T is optionally substituted heteroaryl;
m is 1 or 2;
U is CH2 or NR6 
R6 is selected from the group consisting of hydrogen, optionally substituted alkyl, aralkyl, optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heteroaryl and optionally substituted heterocyclo and COR9;
R9 is selected from the group consisting of hydrogen, optionally substituted alkyl, aralkyl, optionally substituted cycloalkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted aryl, optionally substituted heteroaryl and optionally substituted heterocyclo; and
P1 is an amine protecting group.
CV. The compound of CIV, wherein P1 is selected from the group consisting of t-butoxycarbonyl and benzyloxycarbonyl.
CVI. A compound having Formula XXV




wherein:
T is optionally substituted heteroaryl;
m is 1 or 2;
U is CH2 or NR6 
R6 is selected from the group consisting of hydrogen, optionally substituted alkyl, aralkyl, optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heteroaryl and optionally substituted heterocyclo and COR9;
R9 is selected from the group consisting of hydrogen, optionally substituted alkyl, aralkyl, optionally substituted cycloalkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted aryl, optionally substituted heteroaryl and optionally substituted heterocyclo;
X is selected from the group consisting of hydrogen, optionally substituted alkyl, and aralkyl;
A1 is selected from the group consisting of hydrogen an optionally substituted alkyl; and
P1 is an amine protecting group.
CVII. The compound of CVI, wherein P1 is selected from the group consisting of t-butoxycarbonyl and benzyloxycarbonyl.
CVIII. The compound of CVII having the structure:




The following examples are illustrative, but not limiting, of the method and compositions of the present invention. Other suitable modifications and adaptations of the variety of conditions and parameters normally encountered in clinical therapy and which are obvious to those skilled in the art are within the spirit and scope of the invention.
Example 1
Synthesis of Covalently Constrained Smac Mimetics
General Methods: NMR spectra were acquired at a proton frequency of 300 MHz. 1H chemical shifts are reported with Me4Si (0.00 ppm), CHCl3 (7.26 ppm), CD2HOD (3.31 ppm), or DHO (4.79 ppm) as internal standards. 13C chemical shifts are reported with CDCl3 (77.00 ppm), CD3OD (49.00 ppm), or 1,4-dioxane (67.16 ppm) as internal standards. Optical rotations were measured at room temperature. Compounds of the invention may be purified by reverse phase HPLC (0.1% TFA in water and 0.1% TFA in acetonitrile as the eluent) and isolated as the TFA salt.
General Procedure A (Condensation Between Carboxylic Acid and Amine):
To a solution of the two substrates in CH2Cl2 (20 mg/mL for the minor substrate) was added EDC (1.1 eq per amino group), HOBt (1.1 eq per amino group) and N,N-diisopropylethyl amine (4 eq per amino group) at 0° C. with stirring. The mixture was stirred at room temperature for eight hours and then concentrated. The residue was purified by chromatography to give the product.
General Procedure B (Deprotection of Boc):
To a solution of the substrate in methanol (20 mg/mL) was added a solution of HCl in 1,4-dioxane (4 M, 10-20 eq per Boc). The solution was stirred at room temperature overnight and then condensed to give the product.
Example 2
Synthesis of Smac Mimetic Intermediates
Intermediates in the synthetic pathway for conformationally constrained Smac mimetics may be synthesized using methodology described in Schemes 1-7.




The synthesis of intermediates 5 and 7 is shown in Scheme 1. Compound 2 may be prepared in five steps from pyroglutamic acid 1 according to reported methods (see: (1) Zhang, J.; Xiong, C.; Wang, W.; Ying, J.; Hruby, V., J. Org. Lett., 2002, 4 (23), 4029-4032, (2) Polyak, F. and Lubell, W. D. J. Org, Chem. 1998, 63, 5937-5949, and (3) Tetrahedron Letters 2005, 46, 945-947.) as a mixture of two diastereoisomers with the R form isomer as the major product (ratio is about 4:1). Removal of the Boc group in 2 followed by condensation with N-α-(tert-butoxylcarbonyl)-N-β-(benzoxylcarbonyl)-L-diamino-propionic acid (Boc-Dap(Z)—OH) gave amide 3. Ozone oxidation of the C—C double bond in 3 yielded aldehyde 4. Cleavage of the Cbz group in 4, intramolecular condensation of the resulting amine with the aldehyde group and subsequent reduction of the enamine were realized in one pot to give compound 5 under prolonged reaction times. Alternatively, deprotection of the CBz group of 4, intramolecular cyclization, isolation of the enamine intermediate and reduction provides 5. In this transformation only compound 5 was obtained and there was no detectable formation of its isomer, suggesting that the amino aldehyde from the minor isomer does not cyclize under these conditions.
To a solution of compound 2 (540 mg, 2 mmol) in 20 mL of methanol was added 4 mL of a solution of 4 N HCl in 1,4-dioxane. The solution was stirred at room temperature overnight and then concentrated to give an ammonium salt. To a mixture of this salt in 15 mL of dichloromethane were added 1.17 g (2.4 eq) of Boc-Dap(Z)—OH.DCHA, 460 mg (2.4 mmol) of EDC, 320 mg (2.4 mmol) of HOBt, and 3 mL of N,N-diisopropylethyl amine. The mixture was stirred at room temperature overnight and then condensed. The residue was purified by chromatography to afford compound 3 (YP-348) (580 mg, 59%). 1H NMR (300 MHz, CDCl3, TMS) (major isomer) δ 7.34-7.28 (m, 5H), 5.80-5.77 (m, 1H), 5.59 (m, 1H), 5.36-5.33 (d, J=10.0 Hz, 2H), 5.19-5.01 (m, 4H), 4.67-4.62 (m, 1H), 4.47-4.44 (m, 1H), 3.76-3.74 (s, 1H), 3.74-3.71 (s, 2H), 2.32-2.30 (m, 1H), 2.16-2.12 (m, 1H), 1.99-1.95 (m, 2H), 1.42 (s, 9H); 13C NMR (75 MHz, CDCl3) δ 172.4, 170.5, 156.5, 155.2, 136.4, 134.6, 133.8, 128.3, 127.9, 118.5, 117.1, 80.0, 66.6, 59.7, 58.2, 52.6, 43.4, 29.2, 28.1, 26.6.
O3 was bubbled through a solution of compound 3 (490 mg, 1 mmol) in 20 mL of CH2Cl2 at −78° C. until the color turned to pale blue. O3 was bubbled for 15 min more before air was bubbled to get rid of excessive O3. After adding 3 mL of Et3N, the mixture was warmed to room temperature and stirred for 1 h. The solvent was evaporated and the residue was purified by chromatography to give aldehyde 4 (YP-367) (340 mg, 69%). 1H NMR (300 MHz, CDCl3, TMS) (major isomer) δ 9.78-9.67 (m, 1H), 7.53-7.32 (m, 5H), 5.44 (s, 1/2H), 5.32 (s, 1/2H), 5.15-5.06 (m, 2H), 4.64 (m, 1H), 4.40-4.39 (m, 1H), 3.78-3.76 (s, 3/2H), 3.76-3.74 (s, 3/2H), 3.48-3.42 (m, 3H), 2.78-2.52 (m, 1H), 2.40-2.20 (m, 1H), 2.16 (m, 2H), 2.06-1.89 (m, 1H), 1.44-1.43 (m, 9H); 13C NMR (75 MHz, CDCl3) δ200.3, 199.5, 172.6, 172.2, 170.3, 156.5, 136.4, 128.4, 128.0, 66.7, 59.7, 59.1, 54.3, 52.4, 52.3, 48.4, 43.3, 29.6, 28.2, 21.0.
To a solution of compound 4 (290 mg, 0.6 mmol) in 20 mL of isopropanol was added 0.2 g of 10% Pd/C. The mixture was stirred at room temperature under H2 overnight, filtered through celite and concentrated. The residue was dissolved in dry THF. To this solution was added NaBH(OAc)3 (380 mg, 1.8 mmol). The mixture was stirred at room temperature overnight, diluted with CH2Cl2, washed with brine, dried over Na2SO4 and concentrated. The residue was purified by chromatography to give compound 5 (72 mg, 35%). [α]20D 30.2 (c=1.7, CHCl3); 1H NMR (300 MHz, CDCl3, TMS) δ 5.45 (brd, J=8.0 Hz, 1H), 4.67 (m, 1H), 4.52 (t, J=9.0 Hz, 1H), 4.23 (m, 1H), 3.74 (s, 3H), 3.20 (m, 2H), 2.94 (m, 1H), 2.74 (dd, J=13.6, 10.9 Hz, 1), 2.35 (m, 1H), 2.14 (m, 1H), 1.99 (m, 1H), 1.86-1.74 (m, 3H), 1.66 (m, 1H), 1.43 (brs, 9H); 13C NMR (75 MHz, CDCl3, TMS) δ 173.42, 170.60, 155.16, 79.68, 59.46, 58.39, 54.92, 52.44, 46.72, 37.45, 32.15, 29.64, 28.29, 26.98.
Hydroboration of the C—C double bond in 3 with 9-BBN followed by alkaline oxidation of the resulted borane afforded alcohol 6. Oxidation of 6 with Dess-Martin periodinane furnished a mixture of two aldehydes, which was cyclized in the same procedure as that for compound 5 to give compound 7. Similar to 5, during this transformation only one isomer was obtained.
Analytical data for compound 7: [α]20D−23.2 (c=1.0, CHCl3); 1H NMR (300 MHz, CDCl3, TMS) δ 5.23 (brd, J=8.0 Hz, 1H), 4.79 (m, 1H), 4.65 (dd, J=9.7, 8.2 Hz), 4.22 (m, 1H), 3.74 (s, 3H), 3.02-2.80 (m, 4H), 2.38-1.70 (m, 9H), 1.43 (brs, 9H); 13C NMR (75 MHz, CDCl3, TMS) δ 173.38, 171.59, 155.09, 79.68, 62.03, 59.82, 53.72, 53.15, 52.48, 50.09, 34.66, 34.55, 29.47, 28.31, 27.33.




Analytical data for YP-248P: 1H NMR shows that this compound has two rotamers with a ratio of 2:1. 1H NMR (300 MHz, CDCl3, TMS) δ 7.47-7.44 (m, 1H), 7.38-7.32 (m, 4H), 5.65-5.62 (d, J=8 Hz, 1H), 5.31-5.16 (m, 2H), 4.64-4.60 (m, 1H), 4.51-4.46 (t, J=8 Hz, 1H), 4.24-4.23 (m, 1H), 4.23-4.21 (m, 1H), 3.75 (s, 1H), 3.73 (s, 2H), 3.66-3.63 (m, 1H), 3.63-3.61 (m, 1H), 3.61-3.31 (m, 1H), 2.36-2.34 (m, 1H), 2.11-1.76 (m, 6H), 1.44-1.45 (s, 9H).
Analytical data for amide intermediate: 1H NMR (300 MHz, CDCl3, TMS) δ 5.79 (brd, J=7.0 Hz, 1H), 4.50-4.35 (m, 2H), 4.05 (m, 1H), 3.98-3.85 (m, 2H), 3.70 (s, 3H), 3.32-3.04 (m, 2H), 2.54 (m, 1H), 2.40-2.26 (m, 2H), 2.25-1.60 (m, 6H), 1.39 (s, 9H), 0.98-0.89 (m, 6H); 13C NMR (75 MHz, CDCl3) δ 173.12, 172.52, 168.85, 154.69, 79.80, 59.51, 56.11, 54.38, 53.51, 52.23, 46.18, 42.02, 32.51, 31.12, 28.12, 26.54, 25.81, 22.69, 22.40.




Analytical data for YP-237P: [α]20D−21.5° (c=1.0, CHCl3); 1H NMR (300 MHz, CDCl3, TMS) δ 3.71 (t, J=6.5 Hz, 3H), 3.60 (dd, J=9.0, 5.4 Hz, 1H), 3.11 (m, 1H), 2.05 (m, 1H), 1.95-1.63 (m, 3H), 1.46 (s, 9H), 1.25 (m, 1H), 0.89 (s, 9H), 0.05 (s, 6H); 13C NMR (75 MHz, CDCl3) δ 174.5, 80.8, 61.5, 60.6, 57.5, 38.8, 31.8, 30.4, 28.0, 25.9, 18.2, −5.4; HRMS: calcd. m/z for [M+H]+ 330.2464; found 330.2466.
Analytical data for YP-238P: [α]20D−90.0° (c=1.67, CHCl3); 1H NMR shows this compound has two rotamers with a ratio of 1:1. 1H NMR (300 MHz, CDCl3, TMS) δ 7.28 (m, 5H), 5.59 (m, 1H), 5.35 (m, 1H), 5.20-5.05 (m, 2H), 4.85 (m, 1⁄2H), 4.65 (m, 1⁄2H), 4.46 (m, 1H), 4.35 (m, 1H), 3.80 (m, 1⁄2H), 3.70-3.50 (m, 2H), 3.40 (m, 1H), 3.25 (m, 1⁄2H), 2.32 (m, 1H), 2.20-1.50 (m, 4H), 1.46 (s, 4.5H), 1.44 (s, 4.5H), 1.43 (s, 4.5H), 1.41 (s, 4.5H); HRMS: calcd m/z 558.2791 for [M+Na]+; found 558.2794.
Analytical data for YP-239: [α]20D−51.6° (c=1.67, CHCl3); 1H NMR shows that this compound has two rotamers with a ratio of 2:1. 1H NMR (300 MHz, CDCl3, TMS) δ 9.76 (s, 2/3H), 9.71 (s, 1/3H), 7.40-7.28 (m, 5H), 5.72-5.30 (m, 2H), 5.20-4.95 (m, 2H), 4.90-4.25 (m, 3H), 3.52-3.05 (m, 3H), 2.90-1.60 (m, 4H), 1.50-1.35 (m, 18H); HRMS: calcd m/z 556.2635 for [M+Na]+; found 556.2629.
Analytical data for YP-239P: [α]20D−8.4° (c=0.65, CHCl3); 1H NMR (300 MHz, CDCl3, TMS) δ 5.49 (brd, J=8.1 Hz, 1H), 4.70 (m, 1H), 4.41 (t, J=9.3 Hz, 1H), 4.30 (m, 1H), 3.25-3.18 (m, 2H), 2.89 (m, 1H), 2.75 (dd, J=13.5, 11.1 Hz, 1H), 2.34 (m, 1H), 2.18-1.60 (m, 6H), 1.49 (s, 9H), 1.44 (s, 9H); 13C NMR (75 MHz, CDCl3) δ 171.8, 170.4, 155.2, 81.7, 79.5, 60.6, 58.5, 54.9, 52.3, 46.9, 37.5, 32.1, 28.3, 28.0, 27.0; HRMS: calcd m/z 406.2318 for [M+Na]+; found 406.2317.




Compound 6 can be prepared according to methods reported in the literature (Duggan et al., Org. Biomol. Chem. 3:2287 (2005)) (Scheme 4). Reduction of the alkene and hydrolysis of the benzyl ester gave acid 7.




A compound represented by formula A, wherein m is 1-2, R10a and R10b are independently hydrogen, optionally substituted alkyl, haloalkyl, aralkyl, optionally substituted cycloalkyl, optionally substituted cycloalkenyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted aryl, optionally substituted heteroaryl or optionally substituted heterocyclo, and U has the meaning as described above for Formula I, may be prepared by the method shown in Scheme 5. Briefly, condensation of acid a with ammonia gives primary amide b. Selective transformation of the primary amide to thioamide c can be realized by reaction of b with P4S10 in CH2Cl2 at room temperature. Reaction of c with d, wherein L2 is a leaving group, furnishes e. L2 is a leaving group. In one embodiment, d is an α-bromoketone. Cyclization of e by refluxing in ethanol provides thiozole of Formula A.




A compound represented by Formula B, wherein m is 1-2, R11a, R11b, R11c and R11d are independently hydrogen, optionally substituted alkyl, haloalkyl, aralkyl, optionally substituted cycloalkyl, optionally substituted cycloalkenyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocyclo, halo, nitro, cyano, hydroxy, amino, alkoxy, aryloxy, arylalkyloxy, alkylthio, carboxamido and sulfonamido, R12 is hydrogen, optionally substituted alkyl, haloalkyl, aralkyl, optionally substituted cycloalkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocyclo, and U has the meaning as described above for Formula I, may be prepared as described in Scheme 6. Briefly, condensation of acid a with a substituted diaminobenzene b gives amide c. Cyclization of c by refluxing in AcOH provided compound of Formula B.




A compound represented by Formula C wherein m is 1 or 2, and A1, A2, Z, X, T and U have the meanings as described above for Formula I, may be prepared as shown in Scheme 7. Briefly, removal of the Boc protecting group in a provides amine b. Condensation of b with corresponding Boc-protected amino acid gives amide c. Removal of the Boc protecting group in c affords d. Introduction of the A1 group by substitution of d with an alkyl halide or reductive amination of d with the corresponding aldehyde provides Smac mimetic represented by Formula C.




A compound represented by Formula D wherein m is 1 or 2, and T and R9 have the meanings as described above for Formula I, may be prepared as described in Scheme 8. Briefly, reaction of amine a with carboxylic acid R9CO2H or carboxylic acid chloride R9COCl gives amide b. Removal of the Boc protecting group of b gives a compound represented by Formula D.







A compound represented by Formula E wherein m is 1 or 2, and R9 has the meaning as described for Formula I, may be prepared as described in Scheme 9. Briefly, reaction of acid 1 with 3-bromo-1,2-diamine gives amide 2. Cyclization of 2 in acetic acid gives benzimidazole 3. Suzuki compling with an arylboronic acid (e.g., 2-, 3-, or 4-fluorophenyl boronic acid) gives 4. Boc deprotection of 4 and reaction with L-N-Boc-N-methylalanine gives 5. Cbz deprotection, coupling with R9CO2H, and Boc deprotection gives a compound represented by Formula E.
Example 3




Analytical data for SM-1229: 1H NMR (300 MHz, D2O) δ 7.65-7.50 (m, 2H), 7.21 (s, 1H), 7.18-6.96 (m, 3H), 5.20 (t, J=6.86 Hz, 1H), 4.70 (m, 1H), 4.20 (m, 1H), 3.89 (m, 1H), 2.65 (s, 3H), 2.20-1.70 (m, 4H), 1.70-1.20 (m, 11H); 13C NMR (75 MHz, D2O) δ 173.18, 172.38, 169.49, 153.65, 133.89, 129.09, 128.52, 126.44, 114.79, 60.55, 60.26, 57.20, 50.95, 37.08, 33.03, 32.44, 31.37, 31.15, 25.28, 22.63, 15.72.
Example 4




Analytical data for SM-1235 (major conformer): 1H NMR (300 MHz, D2O) δ 7.72-7.65 (m, 2H), 7.48 (s, 1H), 7.40-7.20 (m, 3H), 5.21 (m, 1H), 5.02 (m, 1H), 4.32 (m, 1H), 3.98-3.20 (m, 5H), 2.55 (s, 3H), 2.35-1.70 (m, 9H), 1.42 (d, J=7.2 Hz, 3H), 0.80-0.62 (m, 6H); 13C NMR (75 MHz, D2O) δ 175.84, 173.60, 169.91, 154.59, 133.96, 129.42, 126.69, 114.57, 71.08, 61.57, 59.80, 57.28, 51.35, 46.62, 42.82, 34.30, 31.62, 22.25, 22.09, 15.61.
Example 5




Analytical data for SM-1237: 1H NMR (300 MHz, D2O) δ 7.45 (d, J=7.5 Hz, 1H), 7.36-7.20 (m, 6H), 71.5 (d, J=7.5 Hz, 1H), 5.13 (m, 1H), 4.72 (m, 1H), 4.33 (m, 1H), 3.80 (m, 1H), 2.51 (s, 3H), 2.45-2.10 (m, 3H), 1.92-1.37 (m, 8H), 1.36 (d, J=7.2 Hz, 3H), 1.15 (m, 1H); 13C NMR (75 MHz, D2O) δ 173.85, 169.76, 153.02, 135.56, 131.66, 129.48, 129.18, 128.72, 128.50, 128.26, 127.10, 113.10, 60.97, 57.15, 55.36, 51.23, 36.59, 32.72, 32.34, 31.31, 29.58, 24.86, 22.80, 15.55.
Example 6




Analytical data for SM-1238 (major conformer): 1H NMR (300 MHz, D2O) δ 7.70 (m, 1H), 7.60-7.40 (m, 7H), 5.40 (m, 1H), 5.15 (m, 1H), 4.55 (m, 1H), 4.03-3.86 (m, 2H), 3.62-3.55 (m, 2H), 3.20 (m, 1H), 2.62 (s, 3H), 2.60-2.20 (m, 4H), 2.15-1.70 (m, 3H), 1.50 (d, J=7.2 Hz, 3H), 1.42 (m, 1H), 0.96 (m, 1H), 0.55 (d, J=7.2 Hz, 3H), 0.36 (d, J=7.2 Hz, 3H).
Example 7




Analytical data for the compound of Example 7: 1H NMR (300 MHz, CDCl3, TMS) δ 7.70-7.35 (m, 6H), 7.32-7.15 (m, 2H), 6.88 (brs, 1H), 5.55 (m, 1H), 5.20 (m, 1H), 4.65 (brm, 1H), 4.42 (m, 1H), 4.15 (m, 1H), 3.25-3.08 (m, 2H), 2.85 (m, 1H), 2.75 (s, 3H), 2.70 (m, 1H), 2.52 (m, 1H), 2.35-1.75 (m, 5H), 1.45 (brs, 9H), 1.35 (d, J=7.0 Hz, 3H); ESI MS (m/z) 561.3 (M+H)+.
Example 8




Analytical data for SM-1257 (major conformer): 1H NMR (300 MHz, D2O) δ 7.70-7.49 (m, 8H), 7.15-7.02 (m, 3H), 6.72-6.62 (m, 2H), 5.47 (m, 1H), 5.15 (m, 1H), 4.75 (m, 1H), 4.59 (m, 1H), 4.10-3.85 (m, 2H), 3.72-3.62 (m, 2H), 3.43 (m, 1H), 2.88 (m, 1H), 2.70 (s, 3H), 2.68-2.25 (m, 4H), 2.20-1.82 (m, 3H), 1.55 (d, J=7.0 Hz, 3H); ESI MS (m/z) 579.3 (M+H)+.
Example 9




Analytical data for SM-1268 (major conformer): 1H NMR (300 MHz, D2O) δ 7.69-7.42 (m, 8H), 6.70-6.52 (m, 4H), 5.48 (m, 1H), 5.20 (m, 1H), 4.75 (m, 1H), 4.62 (m, 1H), 4.09-3.92 (m, 2H), 3.75-3.62 (m, 2H), 3.42 (m, 1H), 2.85 (m, 1H), 2.70 (s, 3H), 2.68-2.25 (m, 4H), 2.18-1.83 (m, 3H); ESI MS (m/z) 597.3 (M+H)+.
Example 10




Analytical data for SM-1270 (major conformer): 1H NMR (300 MHz, D2O) δ 7.82 (m, 1H), 7.70-6.97 (m, 7H), 7.20-7.09 (m, 3H), 6.73-6.65 (m, 2H), 5.49 (m, 1H), 5.23 (m, 1H), 4.75 (m, 1H), 4.53 (m, 1H), 4.10-3.95 (m, 2H), 3.68-3.58 (m, 2H), 3.32 (m, 1H), 2.72 (s, 3H), 2.72-2.55 (m, 2H), 2.52-1.95 (m, 6H), 1.80-1.62 (m, 2H), 1.55 (d, J=7.0 Hz, 3H); ESI MS (m/z) 593.3 (M+H)+.
Example 11




Analytical data for SM-1271 (major conformer): 1H NMR (300 MHz, D2O) δ 7.64 (m, 1H), 7.45-7.20 (m, 6H), 7.05 (m, 1H), 6.65-6.50 (m, 2H), 6.50-6.39 (m, 2H), 5.42 (m, 1H), 5.20 (m, 1H), 4.85 (m, 1H), 4.55 (m, 1H), 4.10-3.90 (m, 2H), 3.65-3.45 (m, 2H), 3.25 (m, 1H), 2.68 (s, 3H), 2.65-2.02 (m, 7H), 1.95-1.70 (m, 1H), 1.55 (d, J=7.0 Hz, 3H), 1.54 (m, 1H); ESI MS (m/z) 611.3 (M+H)+.
Example 12




Analytical data for SM-1306: 1H NMR (300 MHz, D2O) δ 7.70-7.50 (m, 4H), 7.40-7.20 (m, 3H), 6.82-6.58 (m, 4H), 5.47 (m, 1H), 5.18 (m, 1H), 4.75-4.50 (m, 2H), 4.05-3.88 (m, 2H), 3.75-3.65 (m, 2H), 3.50-3.30 (m, 2H), 2.90 (m, 1H), 2.65 (m, 3H), 2.62-1.90 (m, 6H), 1.55 (d, J=7.0 Hz, 3H); ESI MS (m/z) 615.3 (M+H)+.
Example 13




Analytical data for SM-1307: 1H NMR (300 MHz, D2O) δ 7.40 (m, 1H), 7.25-7.15 (m, 3H), 7.05-5.85 (m, 3H), 6.60-6.35 (m, 4H), 5.38 (m, 1H), 5.15 (m, 1H), 4.75 (m, 1H), 4.50 (m, 1H), 4.05-3.80 (m, 2H), 3.70-3.50 (m, 2H), 3.50-3.20 (m, 2H), 2.70 (m, 1H), 2.63 (s, 3H), 2.60-1.70 (m, 6H), 1.55 (d, J=7.0 Hz, 3H); ESI MS (m/z) 615.3 (M+H)+.
Example 14




Analytical data for SM-1308: 1H NMR (300 MHz, D2O) δ 7.38 (m, 1H), 7.30-7.10 (m, 2H), 7.05-6.85 (m, 4H), 6.60-6.35 (m, 4H), 5.35 (m, 1H), 5.15 (m, 1H), 4.75 (m, 1H), 4.42 (m, 1H), 3.95 (m, 1H), 3.75 (m, 1H), 3.60-3.02 (m, 4H), 2.75-2.60 (m, 4), 2.58-1.60 (m, 6H), 4.05-3.80 (m, 2H), 3.70-3.50 (m, 2H), 3.50-3.20 (m, 2H), 2.70 (m, 1H), 2.63 (s, 3H), 2.60-1.70 (m, 6H), 1.55 (d, J=7.0 Hz, 3H); ESI MS (m/z) 615.3 (M+H)+.
Example 15




Analytical data for SM-1316: 1H NMR (300 MHz, D2O) δ 7.70 (m, 1H), 7.55-7.45 (m, 2H), 7.40 (m, 1H), 7.25-7.15 (m, 2H), 5.45 (m, 1H), 5.20 (m, 1H), 4.75 (m, 1H), 4.55 (m, 1H), 4.03-3.90 (m, 2H), 3.70-3.50 (m, 2H), 3.25 (m, 1H), 2.70 (s, 3H), 2.60-1.60 (m, 7H), 1.55 (d, J=7.0 Hz, 3H); 1.50 (m, 1H), 1.02 (m, 1H), 0.55 (d, J=7.2 Hz, 3H), 0.30 (d, J=7.2 Hz, 3H); ESI MS (m/z) 563.3 (M+H)+.
Example 16




Analytical data for SM-1317: 1H NMR (300 MHz, D2O) δ 7.55 (m, 1H), 7.50-7.30 (m, 3H), 7.22 (m, 1H), 7.18-7.05 (m, 2H), 6.98-6.90 (m, 3H), 6.85-6.50 (m, 2H), 5.38 (m, 1H), 5.10 (m, 1H), 4.80 (m, 1H), 4.50 (m, 1H), 4.02-3.80 (m, 2H), 3.75-3.50 (m, 2H), 3.45-3.20 (m, 2H), 2.78 (m, 1H), 2.68 (s, 3H), 2.58-1.75 (m, 6H), 1.55 (d, J=7.0 Hz, 3H); ESI MS (m/z) 597.3 (M+H)+.
Example 17
Binding of Inhibitors to XIAP
A sensitive and quantitative in vitro binding assay using the fluorescence polarization (FP) based method was used to determine the binding affinity of Smac mimetics to XIAP protein (Nikolovska-Coleska et al., Anal. Biochem. 332:261-73 (2004)). For this assay, 5-carboxyfluorescein (5-Fam) was coupled to the lysine side chain of the mutated Smac peptide, AbuRPF-K-(5-Fam)-NH2 (termed SM5F). Another fluorescently tagged Smac mimetic (termed SM-F1) with higher affinities than SM5F was also used.
For determination of the Kd values, fluorescence polarization values were measured using the Ultra plate reader (Tecan U.S., Research Triangle Park, N.C.) in Microfluor 2 96-well, black, round-bottom plates (Thermo Scientific). As one example for the SM-F1 tracer, to each well, SM-F1 (2 nM, 1 nM, and 1 nM for experiments with XIAP-BIR3, cIAP1-BIR3, and cIAP2-BIR3, respectively) and different concentrations of proteins were added to a final volume of 125 μl in the assay buffer (100 mM potassium phosphate, pH 7.5, 100 μg/ml bovine y-globulin, 0.02% sodium azide, Invitrogen, with 4% DMSO). Plates were mixed and incubated at room temperature for 3 hours with gentle shaking to assure equilibrium. The polarization values in millipolarization units (mP) were measured at an excitation wavelength of 485 nm and an emission wavelength of 530 nm. Equilibrium dissociation constants (Kd) were then calculated by fitting the sigmoidal dose-dependent FP increases as a function of protein concentrations using Graphpad Prism 5.0 software (Graphpad Software, San Diego, Calif.).
The Kd values of SM5F to recombinant XIAP BIR3, cIAP-1 BIR3, cIAP-2 BIR3 were determined to be 17.9 nM (Nikolovska-Coleska et al., Anal. Biochem. 332:261-73 (2004)). The Kd values of SM-F1 to recombinant XAIP-BIR3 protein was determined to be 4.7 nM.
In competitive binding experiments using SM5F as the tracer to XIAP BIR3 protein, the tested compound was incubated with XIAP BIR3 protein (30 nM) and SM5F (5 nM) in the assay buffer (100 mM potassium phosphate, pH 7.5; 100 μg/ml bovine gamma globulin; 0.02% sodium azide, Invitrogen). Dose-dependent competitive binding FP experiments were carried out with serial dilutions of tested compounds. For each assay, the bound peptide control containing recombinant XIAP BIR3 protein and SM5F (equivalent to 0% inhibition) and free peptide control containing only free SM5F (equivalent to 100% inhibition) will be included. The polarization values were measured after 3 hrs of incubation when the binding reached equilibrium using an ULTRA READER (Tecan U.S. Inc., Research Triangle Park, N.C.). IC50 values, the inhibitor concentration at which 50% of bound peptide is displaced, will be determined from a plot using nonlinear least-squares analysis. Curve fitting will be performed using GRAPHPAD PRISM software (GraphPad Software, Inc., San Diego, Calif.).
In competitive binding experiments using SM-F1 as the tracer to XIAP BIR3 protein, the tested compound was incubated with XIAP BIR3 protein (10 nM) and SM-1F (2 nM) in the assay buffer (100 mM potassium phosphate, pH 7.5; 100 μg/ml bovine gamma globulin; 0.02% sodium azide, Invitrogen). Dose-dependent competitive binding FP experiments were carried out with serial dilutions of tested compounds. For each assay, the bound peptide control containing recombinant XIAP BIR3 protein and SM-1F (equivalent to 0% inhibition) and free peptide control containing only free SM-1F (equivalent to 100% inhibition) will be included. The polarization values were measured after 3 hrs of incubation when the binding reached equilibrium using an ULTRA READER (Tecan U.S. Inc., Research Triangle Park, N.C.). IC50 values, the inhibitor concentration at which 50% of bound peptide is displaced, will be determined from a plot using nonlinear least-squares analysis. Curve fitting will be performed using GRAPHPAD PRISM software (GraphPad Software, Inc., San Diego, Calif.).
Negative controls containing protein/tracer complex only (equivalent to 0% inhibition), and positive controls containing only free tracers (equivalent to 100% inhibition), were included in each assay plate. FP values were measured as described above. IC50 values were determined by nonlinear regression fitting of the competition curves. The Ki values of competitive inhibitors were calculated using the equation described before (Nikolovska-Coleska et al., Anal. Biochem. 332:261-73 (2004)), based upon the measured IC50 values, the Kd values of the tracer to different proteins, and the concentrations of the proteins and tracers in the competitive assays. Ki values were also calculated using a commonly used equation known in the literature (Huang, X. J. Biomol. Screen. 8:34-38 (2003)).
When tested in the binding assay, Smac mimetics of the present invention exhibited strong binding affinity to XIAP BIR3 protein as illustrated in Table 2 using either SM5F or SM-F1 as the tracer and FIG. 1 using the SM5F as the tracer. These data indicate that these Smac mimetics bind to XIAP with high affinities.










TABLE 2








XIAP BIR3



Compounds
IC50 (nM)



















SM-1238
<200



SM-1237
<200



SM-1235
<200



SM-1299
<5000



SM-1257
<500



SM-1268
<500



SM-1270
<500



SM-1271
<500



SM-1306
<5000



SM-1307
<500



SM-1308
<500



SM-1316
<500



SM-1317
<500










Example 18
Binding of Inhibitors to Other IAP Proteins
In order to test the binding ability of conformationally constrained Smac mimetics to other IAP proteins (cIAP1 and cIAP2) binding assay conditions were developed. The recombinant cIAP1 BIR3 domain (residues 253-363), cIAP2 BIR3 domain (residues 238-349), fused to a His-tag, were used in the binding assays. Competitive binding assays for other IAP proteins are performed similarly as that described for XIAP BIR3. The Kd values of SM5F to recombinant cIAP-1 BIR3 and cIAP-2 BIR3 were determined to be 4.1 nM (Peng et al. J. Med. Chem. 51: 8158-8162 (2008)) and 6.6 nM (Peng et al. J. Med. Chem. 51: 8158-8162 (2008)), respectively. The Kd values of SM-F1 to recombinant cIAP-BIR3, and cIAP2-BIRs proteins were determined to be 1.1 nM and 2.3 nM, respectively.
In competitive binding experiments using SM5F as the tracer to cIAP-1 BIR3 protein, the tested compound was incubated with cIAP-1 BIR3 protein (10 nM) and SM5F (2 nM) in the assay buffer. In competitive binding experiments using SM5F as the tracer to cIAP-2 BIR3 protein, the tested compound was incubated with cIAP-2 BIR3 protein (25 nM) and SM5F (2 nM) in the assay buffer. All other procedures for cIAP-1 BIR3 and cIAP-2 BIR3 proteins were the same as those used for XIAP BIR3 protein competitive assays.
In competitive binding experiments using SM-F1 as the tracer to cIAP-1 BIR3 protein, the tested compound was incubated with cIAP-1 BIR3 protein (3 nM) and SM-F1 (1 nM) in the assay buffer. In competitive binding experiments using SM-F1 as the tracer to cIAP-2 BIR3 protein, the tested compound was incubated with cIAP-2 BIR3 protein (5 nM) and SM-F1 (1 nM) in the assay buffer.
As illustrated in FIG. 2 using SM5F as the tracer, Smac mimetics of the present invention exhibited strong binding affinity to cIAP1 BIR3 protein. Table 3 shows binding affinities of compounds of the invention to cIAP 1 and cIAP2 proteins using either SM5F or SM-F1 as the tracer. These data suggest that compounds of the invention will act as potent inhibitors of cIAP 1 and cIAP2 activity.











TABLE 3








cIAP1 BIR3
cIAP2 BIR3



Compounds
IC50 (nM)
IC50 (nM)




















SM-1238
<100
<100



SM-1237
<100
<100



SM-1235
<1000
<1000



SM-1229
<2000
<2000



SM-1257
<100
<100



SM-1268
<100
<100



SM-1270
<100
<100



SM-1271
<100
<100



SM-1306
<100
<100



SM-1307
<100
<100



SM-1308
<100
<100



SM-1316
<100
<100



SM-1317
<100
<100










Example 19
Cell Growth Inhibition by Conformationally Constrained Smac Mimetics
The effect of the compounds of the present invention on the growth of various cancer cell lines was tested. Cells were seeded in 96-well flat bottom cell culture plates at a density of 3000 cells/well with a tested compound and incubated at 37° C. in an atmosphere of 95% air and 5% CO2 for 4 days. The rate of cell growth inhibition after treatment with different concentrations of the compound was determined using a WST-8 kit (2-(2-methoxy-4-nitrophenyl)-3-(4-nitrophenyl)-5-(2, 4 disulfophenyl)-2H-tetrazolium monosodium salt; Dojindo Molecular Technologies, Inc., Gaithersburg, Md.). WST-8 was added at a final concentration of 10% to each well, and then the plates were incubated at 37° C. for 2-3 hrs. The absorbance of the samples was measured at 450 nm using a ULTRA Tecan Reader (Molecular Device). The concentration of the tested compound that inhibited cell growth by 50% (IC50) was calculated by comparing absorbance in untreated cells and the cells treated with the tested compound.
When tested against the MDA-MB-231 human breast cancer cell line and SK-OV-3 ovarian cancer cell line, compounds of the present invention exhibited strong inhibitory activity as shown in Table 4, suggesting that the compounds are inhibitors of cancer cell growth.











TABLE 4








MDA-MB-231
SK-OV-3



Name
IC50 (μM)
IC50 (μM)




















SM-1235
<3
<3



SM-1237
<3
<3



SM-1238
<3
<3



SM-1229
<100
<100



SM-1257
<3
<3



SM-1268
<3
<3



SM-1270
<3
<3



SM-1271
<3
<3



SM-1306
<3
<3



SM-1307
<3
<3



SM-1308
<3
<3



SM-1316
<3
<3



SM-1317
<3
<3










Example 20
Induction of Cell Death
The ability of SM-1238 to induce cell death was tested in the breast cancer MDA-MB-231 and ovarian cancer SK-OV-3 cell lines (FIG. 3). Cells were treated with SM-1238 for 48 hours and cell viability was determined using the trypan blue exclusion assay. SM-1238 induced cell death in both cell lines in a concentration-dependant fashion.
Example 21
Pharmacokinetics in Rats Following Oral Administration
Following oral administration of SM-1238 (Example 6) to male Sprague-Dawley rats at a dose of 25 mg/kg, the mean±SD values for Cmax, Tmax, AUC(0-∞), and half-life (T1⁄2) were 831±135 ug/L, 2.0±1.7 hrs, 7099±931 μg/L*hr, and 3.7±2.0 hr, respectively (n=3.)
Following oral administration of SM-1268 (Example 9) to male Sprague-Dawley rats at a dose of 25 mg/kg, the mean±SD values for Cmax, Tmax, AUC(0-∞), and half-life (T1⁄2) were 612±16 ug/L, 2.0±0.0 hrs, 6489±965 μg/L*hr, and 4.2±1.0 hr, respectively (n=3)
Following oral administration of SM-1316 (Example 15) to male Sprague-Dawley rats at a dose of 25 mg/kg, the mean±SD values of Cmax, Tmax, AUC(0-∞), and half-life (T1⁄2) were 1528±275 ug/L, 2.0±0.0 hrs, 14304±1968 μg/L*hr, and 6.7±1.2 hr, respectively (n=3).
These experiments show that the compounds of the invention are orally bioavailable.
Having now fully described the invention, it will be understood by those of skill in the art that the same can be performed within a wide and equivalent range of conditions, formulations, and other parameters without affecting the scope of the invention or any embodiment thereof. All patents, patent applications and publications cited herein are fully incorporated by reference herein in their entirety.

## Claims

1. A compound having the following formula:



wherein A1 and A2 are selected from either A1 as methyl and A2 as Hydrogen, or A1 as Hydrogen and A2 as methyl;
wherein X is selected from the group consisting of Hydrogen and methyl;
wherein U is selected from the group consisting of CH2,





and
wherein T is selected from the group consisting of

2. The compound of claim 1, selected from the group consisting of:







or a pharmaceutically acceptable salt thereof.

3. The compound of claim 1, selected from the group consisting of:




or a pharmaceutically acceptable salt thereof.