Infectious Diseases – Weiwen Ying, Lijun Sun, Keizo Koya, Dinesh U. Chimmanamada, Shijie Zhang, Teresa Kowalczyk-Przewloka, Hao Li, Synta Phamaceuticals Corp
Abstract for “Triazole compounds which modulate HSP90 activity”
“The invention concerns substituted triazole compound and compositions comprising substituted Triazole compounds. Further, the invention relates to methods for inhibiting Hsp90 activity in subjects in need thereof. Methods for treating or preventing hyperproliferative disorders such as cancer include administering to the subject either a substituted triazole compounds of the invention or a composition consisting of such a compound.
Background for “Triazole compounds which modulate HSP90 activity”
Although great strides have been made in understanding the genetic abnormalities that lead to malignant cancer cells, the current chemotherapy is not satisfactory and the prognosis remains poor for most patients with this disease. The majority of chemotherapeutic agents target a specific molecular target that is thought to be responsible for the malignant phenotype. The proliferation of cells is controlled by a complex network signaling pathways. Most malignant cancers are caused by multiple genetic abnormalities within these pathways. It is unlikely that a therapy agent that targets one molecular target can cure cancer patients.
“Heat shock proteins (HSPs), are chaperone proteins that are upregulated in response to high temperatures and other environmental stressors, such as ultraviolet light and nutrient deprivation and oxygen deprivation. HSPs are chaperones for other cellular proteins (client proteins). They facilitate the proper folding and repair of client proteins and aid in the refolding misfolded client protein. There are many families of HSPs with their own sets of client proteins. Hsp90 is the most common HSP family, accounting for around 1-2% of proteins in cells that are not under stress but increasing to approximately 4-6% in cells under stress. Hsp90 inhibition results in the degradation of its client proteins through the ubiquitin protasome pathway. Hsp90’s client proteins are, unlike other chaperone proteins. They include protein kinases and transcription factors that aid in signal transduction. A number of Hsp90’s client proteins have been implicated in the development of cancer. Below are examples of Hsp90 client protein that have been implicated with the progression of cancer.
Her-2 is a transmembrane Tyrosine Kinase Cell Surface Growth Factor Receptor. It is found in normal epithelial cell. Her2 contains an extracellular domain that interacts and transmits extracellular growth signals to the cell’s nucleus. A high level of Her2 expression is associated with poor prognosis in many malignancies such as breast cancer and ovarian cancer.
“Akt kinase, a serine/threonine-kinase that is a downstream effector molecular of phosphoinositide 3kinase, is involved in protecting cells from apoptosis. Because it stimulates cell proliferation, Akt kinase may be involved in the development of cancer.
“Cdk4/cyclinD complexes are involved with phosphorylation retinoblastoma proteins which is an essential step in the progression of cells through the G1 phase. The half-life of newly synthesized Cdk4 is decreased by disrupting Hsp90 activity.
The Raf family of protooncogenes (A, B, and C-raf), was first discovered when C-raf was identified (raf-1). This was due to its homology in v-raf which is the transform gene of the mouse-sarcomavirus 3611. Later, A-raf, a transforming gene from the Mill Hill No. avaian retrovirus, was found by screening a cDNA database under low stringency conditions with a v?raf probe. B-raf, a homology to C-Rmil was also discovered. 2. The Ras/Raf/MEK/ERK pathway is mediated by the Raf protein family, also known as the?MAP kinase pathways. (MEK stands to?MAPK/ERK Kinase). ERK stands to?extracellularly regulated kinases? It has been linked to the development and progression of many cancers by upregulating cell division and proliferation. All raf proteins can activate the MAP kinase pathway. B-raf activates this pathway more effectively than A-raf and C-raf. Mutations in the gene that encodes B-raf are also more common in cancer. B-raf mutations were found in 60 to 70% of malignant melanomas and 35% to 69% in papillary thyroid carcinoma. They also occur in 35% to 16% of colon carcinomas, 35% to 69% in papillary thyroid caricinoma, 35% to 69% in low-grade ovarian cancer, 4% to 15% in colon cancer, 35% to 48% in head and neck squamous cells carcinoma, 35% to 6.8% in head and neck squamaromaf, cholangiocarcinomaroma, in breast cancer, and cholangiocarcinoma. The majority of B-raf mutations found in human cancers occur in the kinasedomain and cluster in exons 11, 15 and 16. These genes contain several regulatory phosphorylation site (S446, 5447 D448, D449 D449, D449, D449, S599 and S602). (Beeram, and colleagues, Journal of Clinical Oncology, 23(27), 6771-6790. T1799A is the most common mutation. It accounts for more that 80% of BRAF gene mutations and leads to a V600E mutation. V600E was previously known as V599E. The gene mutation was originally called T1796A. This was due to a mistake with the GenBank nucleotide sequence, NM 004333. The correct GenBank sequence is: NT 007914. It designates the protein mutant as V600E, and the gene mutation at T1799A. This will be the corrected numbering. This mutation mimics phosphorylation of B-raf’s activation segment. It inserts a negatively charged residue close to two activating sites for phosphorylation, T599, and 5602. This causes constitutively active B raf in an independent Ras-dependent manner. “Endocrine-Related Cancer (2005) 12:245-262.
The Hsp90 inhibitor 17AAG has been shown in cancer cells to stimulate B-raf’s degradation. Mutant forms of B raf are more susceptible to being degraded than the wild type. When 17AAG was applied to A375, a melanoma cell that has the V600E mutation, B-raf degraded faster than in CHL cells that contained wild-type B-raf. Other B-raf mutations, such as V600D,G469A and G469E, were found to degrade faster than wild type B.raf when they were transfected into COS cell lines. B-raf mutants E586K or L597V did not show any signs of degradation after cells were treated with 17AAG. It is believed that B-raf mutants E586K and L597V are clients of Hsp90. Mutated forms of B -raf, however, are more dependent upon Hsp90 for folding and stability. (Dias, et al., Cancer Res. (2005), 65(23): 10686-10691). Raf-1, a MAP 3-kinase(MAP3K), can activate the serine/threonine-specific protein kinases ERK1 or ERK2. Activated ERBs play an important part in controlling gene expression that is involved in cell division, cell differentiation, and cell migration.
Anaplastic large-cell Lymphoma (ALCL), a non-Hodgkin?s lymphoma, is characterised by the expression CD30/Ki-1. ALCL is normally caused by T-cells. However, some cases may have a null or B-cell phenotype. Diffuse large B-cell lymphomas are cases that arise from B cells. Around 60% of ALCL cases that express CD30/Ki-1 also have the chromosomal transfer t(2:5)(p23.q35), which fuses the nucleophosmin gene (NPM/B233) to the anaplastic lymphomakine (ALK) and produces the oncogenetic protein NPM-ALK with activity. ALK rearrangements were observed in specific types of ALCL: 1) 30% to 50% of pleomorphic ALCL; 2) more than 80% for monomorphic ALCL; 3) 75%?100% of small-cell cases; and 4) 60%?100% of lymphohistiocytic ALCL. NPM-ALK can transform fibroblasts, hematopoietic and primary bone marrow cells. It is believed to activate phosphatidylinositol-3 kinase (PI-3 kinase), which protects against apoptosis and stimulates mitosis via the RAS pathway. (Duyster, et al., Oncogene (2001), 20:5623-5637). NPM?ALK has been shown associate with Hsp90. Incubation of NPM?ALK-expressing ALCL cells with 17AAG of the benzoquinone isamycin has been shown not to disrupt this association. This can lead to increased degradation of NPM?ALK and cell cycle arrest. (Georgakis, et al., Exp. Hematology (2006), 34(12):1670-1679; Bonvini, et al., Cancer Research (2002), 62:1559-1566).”
“V-src is the Rous Sarcoma virus’s transforming protein. It is an example of an oncogene that induces cellular mutation (i.e. tumorogenesis) through non-regulated kinase activities. Hsp90 can interact with v-scr to inhibit its destruction.
“Hsp90 is necessary to maintain steroid hormone receptors at a conformation capable binding high affinity hormones with high affinity. The treatment of hormone-associated malignancies like breast cancer, such as Hsp90 inhibition, is expected to prove useful.
“P53 is a tumor suppressor protein which causes cell cycle arrest, and apoptosis. About half of all cancers are caused by mutations in the p53 gene. This makes it one of the most prevalent genetic alterations in cancerous cells. A poor prognosis is also associated with p53 mutation. Hsp90 interacts with wild-type P53, but mutant p53 forms a stronger association than wild-type Hsp53 due to its misfolded conformations. The mutated protein is protected from normal proteolytic degradation by a stronger interaction with Hsp90, which prolongs its half-life. A heterozygous cell for wild-type and mutated p53 will have mutant p53 degraded by inhibition of Hsp90’s stabilizing effect. This restores normal transcriptional activity for wild-type.
“Hif-1? Hypoxia-inducible transcription factors that are up-regulated in low oxygen conditions. Normal oxygen conditions, Hif-1 is degraded. Hif-1 is a tumor suppressor protein that associates with Von Hippel-Lindau? (VHL). This inhibits Hif-1’s ability to associate with VHL. Hif-1 to accumulate and form complexes with it? To form an active transcription complex which associates with hypoxia response elements to activate transcription of vascular epithelial growth factor. An increase in Hif-1? Increased Hif-1 is associated with poor prognosis and increased metastasis.”
There are two types of PKs. The protein tyrosine kinases, (PTKs), catalyze phosphorylation tyrosine kinase residues and the serine/threonine kinases kinases(STKs), catalyze phosphorylation threonine residues. The receptor tyrosine-kinases are growth factor receptors that have PTK activity. The family of receptor tyrosine-kinases is tightly controlled enzymes. Abnormal activation of different members of this family is a hallmark of cancer. There are subgroups within the receptor tyrosine-kinase family that share similar sequence and structural similarities.
“Epidermal growth factor receptor (EGFR), is a member the type 1 subgroup receptor tyrosine kinase families of growth factor receptors. These receptors play crucial roles in cellular differentiation, growth, and survival. These receptors are activated by specific ligand binding, which results in homo- or heterodimerization of receptor family members and subsequent autophosphorylation. EGFR is bindable to epidermal growth factors (EGF), transforming Growth Factor (?). (TGF?), amphiregulin, and other viral growth factors. Activating EGFR triggers a series of intracellular signaling pathways that are involved in both cellular proliferation (the ras/raf/MAP kinase pathway), and survival (the PI3 kinase/Akt pathway). Several members of this family, including HER2 and EGFR, have been implicated directly in cellular transformation.
“Many human malignancies are linked to aberrant or excessive expression of EGFR, and/or its specific ligands” (Gullick, Br. Med. Bull. (1991), 47.87-98; Modijtahedi, Int. J. Oncol. (1994), 4:277-96; Salomon, et al., Crit. Rev. Oncol. Hematol. (1995); 19;183-232. The entire teachings from each of these references are included herein by reference. Overexpression or aberrant EGFR has been linked to adverse prognosis for a variety of human cancers including head, neck, breast, colon and prostate (e.g. NSCLC, adenocarcinoma, squamous lung carcinoma, and squamous cell carcinoma), ovaries, gastrointestinal (gastric, colon and pancreatic), renal cancer, bladder cancer, glioma and gynecological cancers and prostate cancer. Overexpression of tumor EGFR can be associated with poor prognosis and chemoresistance in some cases (Lei et al. Anticancer Res. (1999), 19:221-8; Veale, et al., Br. J. J.
Gefitinib is a chemotherapeutic drug that inhibits EGFR activity. It has been shown to be very effective in a subset lung cancer patients with mutations in the tyrosine kinasedomain of EGFR. These mutants showed two to three times more activity when EGF was present than wild-type EGFR. The cells also absorbed wild type EGFR and it was down-regulated within 15 minutes. However, mutant EGFR continued to activate for as long as three hours. (Lynch, et. al., The New England Journal of Medicine (2006) 350:2129-2139; the complete teachings of which are incorporated by reference herein).
Another type of cancer is called gliomas. It is caused by amplification or mutation of the EGFR genes. A deletion of exons 2–7 results in a truncated EGFR form in which the amino acids 6-273 and a single glycine atom are replaced. This is one of the most common mutations of the EGFR genes. This mutation is known as EGFRvIII, and it is found in approximately half of all glioblastomas. EGFRvIII cannot bind TGF or EGF. It has constitutive, ligand independent tyrosinekinase activity. Hsp90 copurifies with EGFRvIII, indicating that Hsp90 interacts with EGFRvIII. Hsp90 inhibitor geldanamycin was able decrease the expression of EGFRvIII. This indicates that Hsp90 interacts with EGFRvIII. (Lavictoire, et.al., Journal of Biological Chemistry (2003) 278(7):5292-599; the complete teachings of which are incorporated in this reference). These results show that Hsp90 inhibition is an effective strategy to treat cancers associated with inappropriate EGFR activity.
“The type III group of receptor tyrosine kinases includes platelet-derived Growth Factor (PDGF) receptors, PDGF receptors alpha, beta, colony-stimulating factors (CSF-1R), c-Fms), Fms -like tyrosine kinase FLT3 and stem cell factor receptor c-kit. F1LT3 is expressed primarily on immature human hematopoietic stem cells and regulates their survival and proliferation.
Hematologic cancers are also called hematopoietic or hematologic malignancies. They include leukemia and lymphoma, as well as cancers of blood or bone marrow. Acute myelogenous (AML), a clonal hematopoietic, stem cell leukemia, is about 90% of acute leukemias in adults. It has an incidence rate of 3.9 per 100,000. (See Lowenberg et. al., N. Eng. J. Med. 341: 1051-62 (1999) and Lopesde Menezes, et al, Clin. Cancer Res. (2005), 11(14),:5281-5291. The enter teachings from both references are incorporated here by reference. Although chemotherapy can lead to complete remissions of AML, the long-term survival rate is only about 14%. There are approximately 7,400 AML related deaths each year in the United States. A majority of AML blasts are wild-type FLT3 with 25% to 35% expressing FLT3 kinase receiver mutations that result in constitutively active FLT. AML patients have two types of activating mutations: point mutations in the activating loop and internal tandem duplications. Patients with AML who have FLT3 mutations (ITDs) are indicative of poor survival rates. In patients in remission FLT3 mutations are the most detrimental factor in the relapse rate. 64% of patients with FLT3 mutations relapse within five years. Current Pharmaceutical Design (2005), 11/3449-3457. The entire teachings are included herein by reference. Clinical studies have shown that FLT3 mutations are predictive of a poor prognosis for AML patients. They may also be important in the development and maintenance the disease.
Mixed Lineage Leukemia (MLL), which involves translocations of chromosome 11, band q23 (11q23), occurs in approximately 80% of infants with hematological malignancies, and 10% in adult acute leukemias. Certain 11q23 translocations have been shown to be critical for immortalization of hematopoietic precursors in vitro. However, leukemia can only develop from a secondary genotoxic event. The strong correlation between FLT3 expression and MLL fusion gene gene expression is evident. FLT3 is the most frequently overexpressed gene of MLL. It has also been demonstrated that activating FLT3 and MLL fusion gene expression can induce acute leukemia, with a brief latency period (see Ono et al. J. of Clinical Investigation 2005, 115:919-929; the entire teachings of which have been incorporated herein). It is thought that FLT3 is involved in the maintenance and development of MLL (see Armstrong et al. Cancer Cell 2003, 3:73-183).
“The FLT3?ITD mutation can also be found in approximately 3% of adult myelodysplastic cases and some cases acute lymphocyticleukemia (ALL). (Current Pharmaceutical Design, 2005, 11:3449-3457).
“FLT3 was shown to be a client protein for Hsp90. 17AAG, a benzoquinone-ansamycin antibiotic that inhibits Hsp90 activation, has been shown not to interfere with the association of Flt3 and Hsp90. Treatment with 17?AAG was shown to inhibit the growth of leukemia cells that have either FLT3-ITD or wild-type FLT3 mutations. (Yao et al. Clinical Cancer Research 2003, 9:4483-4493; all teachings are included herein by reference).
“c-Kit, a membrane type III receptor protein Tyrosine Kinase that binds Stem Cell Factor to its extracellular region, is an example of a membrane-type III receptor protein tyrosinekinase. c-Kit is essential for normal hematopoiesis and tyrosinekinase activity. Mutations in c-kit may lead to ligand-independent Tyrosine Kinase Activity, Autophosphorylation and uncontrolled cell proliferation. A variety of pathological states have been linked to c-Kit activation and/or aberrant expression. Evidence for the contribution of cKit to neoplastic disease includes evidence of its association with leukemias, mast cell tumors, small-cell lung cancer, testicular carcinoma, and certain cancers of central nervous system and gastrointestinal tract. C-Kit has also been linked to neurofibromatosis and neuroectodermal sarcomas in the female genital tract. (Yang et al., J Clin Invest. (2003), 112:1851-1861; Viskochil, J Clin Invest. (2003), 112;1791-1793. The entire teachings from each of these references are included herein by reference. Hsp90 inhibitor 17AAG has been shown that c-Kit is a client protein for Hsp90. Kasumi-1 cells are an acute myeloidleukemia cell line with a mutation in the c-kit gene.
“c-Met” is a receptor Tyrosine Kinase, which is a client protein for Hsp90. It is encoded in the Met protooncogene. HGF (also known as scatter factor (SF), is the natural ligand for c-Met and causes a range of cellular responses including proliferation, survival and wound healing. It also triggers morphogenetic branching and morphogenesis. (Ma et.al., Cancer and Metastasis Reviews (2003) 22: 309-325. Although c-Met, and HGF can be found in many tissues, their expression is usually restricted to epithelial cells and mesenchymal cells, respectively. HGF and c-Met are essential for mammalian development. They have been shown to play an important role in cell migration, cell proliferation, survival, and organization 3D tubular structures (e.g. renal tubular cells, gland formation). In many human cancers, tumor growth, dissemination, and invasion can be caused by dysregulation of cMet and/or HGF. HGF and c-Met are high-expressed in many cancers, and their expression is associated with poor prognosis. (Christensen, and al., Cancer Research 2003, 63:7345-7355). c-Met receptor mutations were found in a variety of cancers, including ovarian cancer, childhood hepatocellular cancer, metastatic head & neck squamous cells carcinomas, esophageal carcinoma and gastric cancer. Met gene amplification, c-Met overexpression has been linked to non-small-cell lung cancer (NSCLC), small-cell lung cancer (SCLC), and colorectal carcinoma. The Tpr/Met fusion proteins has also been found in gastric cancer and osteogenic sarcoma. Multiple kidney tumors can be caused by germine mutations activating c-Metkinase. Many studies have shown that HGF and c-Met expression are linked to the progression of various types of cancer, including lung, colon and breast, prostate, liver and pancreas.
Gleevec’s success in targeting RTKs that are dysregulated by human cancers has been illustrated by its successes in targeting Bcr -Abl in chronic meelogenous Leukemia, c-Kit and gastroinstinal stromal tumours, Herceptin in Her-2-overexpressing breast cancers and Iressa in selected NSCLC with dysregulated EGFR. There is strong evidence to support the use of c-Met for treatment of human cancers. Several small drug molecules that inhibit c?Met are currently being developed. Therapies that target specific RTK can be effective in treating cancer, but they eventually fail because of additional mutations that allow RTK activity to continue with the drug. Moreover, SU11274, a selective cMet inhibitor, is highly effective against wild-type cMet as well as some mutants. However, it has not been proven to be effective against other cMet mutants (Berthou et al., Oncogene (2004) 23:5387-5393). It is therefore necessary to find new anticancer therapies that inhibit cMet’s expression or activity via a different mechanism from those that inhibit cMet directly.
“BCR-ABL, an ocoprotein that has tyrosinekinase activity, has been associated with chronic lymphocytic and acute leukemia. It also affects patients with acute myelogenous and acute lymphocytic lesions (ALL). The BCR-ABL cancer gene has been detected in approximately 2% of AML patients, 20 percent of ALL adults, 5% of ALL children, and at least 90-95% CML patients. Translocation of gene sequences from chromosome 9’s c-ABL protein tyrosine kinase into chromosome 22 results in the BCR-ABL, or Philadelphia chromosome. It has been demonstrated that the BCR-ABL gene can produce at least three alternative Chimeric Proteins, p230 Bcr -Abl p210 Bcr -Abl and p190 Bcr -Abl. These proteins have unregulated Tyrosine Kinase Activity. CML is associated most frequently with the p210BcrAbl fusion proteins, while ALL is associated more often with the p190Bcr-Abl. Bcr-Abl is also associated with various other hematological malignancies, including CML, lymphomas, myelomonocytic Leukemia, lymphomas, and erythroid Leukemia.
Studies have shown that BcrAbl activity and expression can be reduced to treat BcrAbl-positive lesions. Agents such as As2O3 that lower Bcr Abl expression have been proven to be very effective in fighting Bcr abl leukemias. Imatinib, also known as STI571 or Gleevic, inhibits Bcr Abl tyrosine kinase activation and induces differentiation and apoptosis. This causes the eradication in vivo and vitro of Bcr -Abl positive cells. Imatinib treatment is usually effective in inducing remission in patients with CML who are in the chronic phase as well as those in blast crises. In many cases, especially those who had been in blast crises before remission, the remission does not last because the Bcr/Abl fusion protein evolves mutations that make it resistant to Imatinib. ”
“Bcr fusion proteins are composed of Hsp90 complexes and are rapidly destroyed when Hsp90 action is blocked.” It has been demonstrated that geldanamycin is a benzoquinone-ansamycin antibiotic which disrupts the association between Bcr?Abl and Hsp90 results in proteasomal destruction of Bcr?Abl cells.
Mutational analysis has shown that Hsp90 is essential for normal eukaryotic cell survival. Hsp90 overexpression in cancer cells suggests that Hsp90 may play a critical role in cancer cell survival. Cancer cells might also be more sensitive than normal cells to Hsp90 inhibition. Cancer cells are known to have high levels of overexpressed and mutated oncoproteins, which depend on Hsp90 to fold. Hsp90 may also be important for tumor survival because of the hostile environment in which tumor cells are found. Hsp90 inhibition causes simultaneous inhibition of many oncoproteins as well as hormone receptors. This makes it a desirable target for anti-cancer agents. Clinical trials have shown that benzoquinone and ansamycins, which are natural products that inhibit Hsp90 in mice, can show therapeutic activity.
“Benzoquinone ansamycins and their derivatives are promising, but they have a few limitations. They are difficult to formulate because of their low oral bioavailability and limited solubility. They are also metabolized by polymorphic cytochrome CYP3A4 which makes them a substrate for the P-glycoprotein Export Pump, which is involved in multidrug resistance. There is a need for new therapies that improve the prognosis and overcome the limitations of current anti-cancer drugs.
HSPs are extremely conserved, from microorganisms all the way to mammals. Both the host and the pathogen increase HSP production when they infect a host. The infection process appears to have many roles for HSPs. Hsp90, for example, has been shown to be involved in the pathways that are responsible for the uptake and/or death of bacteria in phagocytic cell linings. Yan, L. et al., Eukaryotic Cell, 567-578, 3(3), 2004. Hsp90 is also essential for the uptake binary actin ADPribosylating toxins into eukaryotic cell. Haug. Infection and Immunity 12, 3066-3068. 2004. Hsp90 was also identified to play a role in viral proliferation in many viruses, including the influenza virus, vacciniavirus, herpes simplex virus types I and HIV-1 virus. Momose, F, et al., J. Biol. Chem., 45306-45314, 277(47), 2002; Hung, J., et al., J. Virology, 1379-1390, 76(3), 2002; Li, Y., et al., Antimicrobial Agents and Chemotherapy, 867-872, 48(3), 2004; O’Keefe, B., et al., J. Biol. Chem., 279-287, 275(1), 2000.”
Opportunistic fungal infections have been a growing problem, especially in immunocompromised patients. Hsp90 is implicated in the development of antifungal drug resistance in fungi. Cowen, L. et al., Eukaryotic Cell, 2184-2188, 5(12), 2006; Cowen, L. et al., Science, 309:2185-2189, 2005.”
“The present invention contains compounds that inhibit Hsp90 activity and can be used in the treatment proliferative diseases such as cancer.”
“In one embodiment, this invention provides compounds represented as structural formula (I).”
“wherein:”
“In one embodiment, of the compounds represented by formula (I), the compound is not 3-hydroxy-4-(5-mercapto-4-(naphthalen-1-yl)-4H-1,2,4-triazol-3-yl)phenyl dihydrogen phosphate.”
“Compounds shown in Table 1 and compounds of any formula therein, as well as tautomers or pharmaceutically acceptable salts of any formula, or compounds of any formula thereof, or solvates, chlorates, clathrates or hydrates, polymorphs, or prodrugs thereof, inhibit Hsp90’s activity and facilitate the degradation of Hsp90 clients proteins. Hsp90 is essential for normal eukaryotic cell survival. Hsp90 may be overexpressed in some tumor types, which suggests that it may play an important role in the survival and growth of cancer cells. Cancer cells might also be more sensitive than normal cells to Hsp90 inhibition. The compounds in Table 1, or any compounds of any formula, or tautomers or pharmaceutically acceptable salts, compounds, clathrates and hydrates, polymorphs, or prodrugs thereof are all useful for treating proliferative disorders like cancer.
Although chemotherapeutic drugs initially cause tumor regression in some cases, the majority of agents currently being used to treat cancer only target one pathway for tumor progression. In many cases, tumors that have been treated with multiple chemotherapeutic drugs develop multidrug resistance, and stop responding to treatment. Hsp90 inhibition has the advantage of inhibiting Hsp90’s activity. Many of its client proteins are protein kinases and transcription factors involved with signal transduction. This can lead to cancer progression. Inhibiting Hsp90 can be used to block multiple pathways that lead to tumor progression. The combination of Hsp90 inhibitors of the invention with other chemotherapy agents is more likely than any other therapies to cause tumor regression or elimination. It also makes it less likely that the tumor will develop multidrug resistance.
“A description of preferred embodiments is given below.”
“The present invention includes compounds described herein, and uses of those compounds for inhibiting Hsp90 activity as well as treatment of proliferative disorders such cancer. The invention includes compounds that can be used to stop or slow down the growth of cancerous cell or reduce or eliminate the number of such cells in a subject. Preferably, the subject is a mammal.
“In some embodiments, the compounds can be combined with other chemotherapy agents. This may reduce or prevent the development of multidrug resistant cancerous cell in mammal.” The compounds of this invention could allow for a lower amount of a second chemotherapy agent to be given to a mammal. This is because the invention compounds should prevent the development of multidrug resistant cells.
“The compounds of the invention may be used in certain embodiments to block, occlude or disrupt blood flow in the neovasculature.”
“In other embodiments, compounds of the invention may be used to treat or inhibit angiogenesis in a subject who is in need.”
“The invention also covers compounds that inhibit topoisomerase II activity.”
“The invention also relates the discovery that treating cells such as peripheral blood mononuclear cell (PMBCs), which have been stimulated by an inflammatory stimuli such as INF/LPS or SAC with an Hsp90 inhibit reduces expression of GR and the production of inflammatory cytokines.
“The invention also includes compounds that inhibit Hsp90’s activity and which are useful in treating or preventing infections.”
“In another embodiment, this invention pertains to a method for treating or preventing fungal drug resistant in mammal that is in need of such treatment.” This method involves administering an Hsp90 inhibitor to the mammal.
“In another embodiment, this invention concerns methods for administering a dosage solution containing compounds of the invention to a mammal.”
“A. TERMINOLOGY”
“Unless otherwise stated, the following terms are used herein:
“The term ‘alkyl’ as used herein means: “A saturated straight chain or branching non-cyclic hydrocarbon with between 1 and 10 carbon atoms. Representative saturated straight chain alkyls include methyl, ethyl, n-propyl, n-butyl, n-pentyl, n-hexyl, n-heptyl, n-octyl, n-nonyl and n-decyl; while saturated branched alkyls include isopropyl, sec-butyl, isobutyl, tert-butyl, isopentyl, 2-methylbutyl, 3-methylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2-methylhexyl, 3-methylhexyl, 4-methylhexyl, 5-methylhexyl, 2,3-dimethylbutyl, 2,3-dimethylpentyl, 2,4-dimethylpentyl, 2,3-dimethylhexyl, 2,4-dimethylhexyl, 2,5-dimethylhexyl, 2,2-dimethylpentyl, 2,2-dimethylhexyl, 3,3-dimtheylpentyl, 3,3-dimethylhexyl, 4,4-dimethylhexyl, 2-ethylpentyl, 3-ethylpentyl, 2-ethylhexyl, 3-ethylhexyl, 4-ethylhexyl, 2-methyl-2-ethylpentyl, 2-methyl-3-ethylpentyl, 2-methyl-4-ethylpentyl, 2-methyl-2-ethylhexyl, 2-methyl-3-ethylhexyl, 2-methyl-4-ethylhexyl, 2,2-diethylpentyl, 3,3-diethylhexyl, 2,2-diethylhexyl, 3,3-diethylhexyl and the like. The term “C1-C6alkyl” refers to a saturated straight chain alkyl. A saturated straight chain or branched, non-cyclic hydrocarbon with between 1 and 6 carbon atoms. The C1-C6 representative alkyl groups shown are those with between 1 and 6 carbon atoms. You can substitute any of the alkyl groups in compounds of this invention with one or more substituents.
“Alkenyl” is the term used herein. “A saturated straight chain, branched non-cyclic carbon having between 2 and 10 carbon atoms with at least one carbon carbon double bond. Representative straight chain and branched (C2-C10)alkenyls include vinyl, allyl, 1-butenyl, 2-butenyl, isobutylenyl, 1-pentenyl, 2-pentenyl, 3-methyl-1-butenyl, 2-methyl-2-butenyl, 2,3-dimethyl-2-butenyl, 1-hexenyl, 2-hexenyl, 3-hexenyl, 1-heptenyl, 2-heptenyl, 3-heptenyl, 1-octenyl, 2-octenyl, 3-octenyl, 1-nonenyl, 2-nonenyl, 3-nonenyl, 1-decenyl, 2-decenyl, 3-decenyl and the like. Optionally, alkenyl groups can be substituted with one or several substituents.
“Alkynyl” is the term used herein. “A saturated straight chain or branched, non-cyclic hydrocarbon with between 2 and 10 carbon atoms and at least one carbon-carbon triple bonds. The following are examples of straight chains and branched alkyls: 1-butyl; 2-butyl; 1-pentynyl; 2-pentynyl. 3-methyl-1-butynyl. 4-pentynyl. 1-octynyl. 2-octynyl. 7-octynyl. 1-nonyl. 2-decynyl. 8-nonyl. 1-decynynynynyl. Optionally, alkyl groups can be substituted with one or several substituents.
“Cycloalkyl” is the term used herein. “A saturated, mono- or multicyclic alkyl radical with between 3 and 20 carbon atoms. Representative cycloalkyls include cyclopropyl, 1-methylcyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl, -cyclodecyl, octahydro-pentalenyl, and the like. Optionally, cycloalkyl groups can be substituted with one or several substituents.
“As used herein the term “cycloalkenyl” means: “A mono- or poly-cyclic, non-aromatic alkyl-radical that has at least one carbon carbon double bond in the cycleic system and between 3 to 20 carbonatoms. Representative cycloalkenyls include cyclopentenyl, cyclopentadienyl, cyclohexenyl, cyclohexadienyl, cycloheptenyl, cycloheptadienyl, cycloheptatrienyl, cyclooctenyl, cyclooctadienyl, cyclooctatrienyl, cyclooctatetraenyl, cyclononenyl, cyclononadienyl, cyclodecenyl, cyclodecadienyl, 1,2,3,4,5,8-hexahydronaphthalenyl and the like. Optionally, cycloalkenyl groups can be substituted with one or several substituents.
“Haloalkyl” is the term used herein. An alkyl group in which one or more hydrogen radicals are replaced with a halo group. Each halo group is selected independently from?F?Cl??Br and?I. The term “halomethyl” is used. The term “halomethyl” refers to a methyl where one or more hydrogen radical(s), has been replaced by a Halo group. Examples of haloalkyl group include bromomethyl and trifluoromethyl.
“Alkoxy” is the term used herein. An alkyl group that is linked to another moiety by an oxygen linker.
“A ‘haloalkoxy’ is a term that means “as used herein.” An oxygen linker is a way to attach haloalkyl groups to other moiety.
“An?aromatic-ring” is the term used herein. Or?aryl? A hydrocarbon monocyclic, polycyclic, or polycyclic radical that has at least one aromatic ring. Examples of suitable aryl groups include, but are not limited to, phenyl, tolyl, anthracenyl, fluorenyl, indenyl, azulenyl, and naphthyl, as well as benzo-fused carbocyclic moieties such as 5,6,7,8-tetrahydronaphthyl. Optionally, aryl groups can be substituted with one or several substituents. One embodiment of the aryl group is monocyclic, and the ring contains 6 carbon atoms. This is called?(C6)aryl.
“Aralkyl” is the term used herein. An aryl group that has been attached to another group by a C1-C6 alkylene group. Aralkyl groups that are representative include naphth-3, 2-phenylmethyl, benzyl and other aryl groups. Optionally, aralkyl groups can be substituted with one or several substituents.
“The term ‘alkylene? as used herein is the following: Refers to an alkyl group with two points of attachment. The term “C1-C6alkylene” is used. An alkylene group with one to six carbon atoms refers to it. Prefer straight chain (C1-C6) alkylene groups. Methylene (?CH2?) is a non-limiting example of an alkylene group. ), ethylene (?CH2CH2? ), n-propylene (?CH2CH2CH2? ), n-propylene (?CH2CH2CH2CH2? ), and other similar. Optionally, Alkylene groups can be substituted with one or several substituents.”
“Heterocyclyl” is the term used herein. “Heterocyclyl” can be used to refer to a monocyclic (typically with 3- to 10 members) or polycyclic (7- to 20-members). It is either a saturated or unsaturated non-aromatic rings. A heterocycle with 3 to 10 members can have up to 5 heteroatoms, while a heterocycle with 7 to 20 members can have up to 7 heteroatoms. A heterocycle typically has at least one carbon-atom ring member. Each heteroatom can be selected independently from nitrogen. This can be oxidized (e.g. N(O), quaternized, oxygen, and sulfur. Any heteroatom or carbon atom can attach the heterocycle. Representative heterocycles include morpholinyl, thiomorpholinyl, pyrrolidinonyl, pyrrolidinyl, piperidinyl, piperazinyl, hydantoinyl, valerolactamyl, oxiranyl, oxetanyl, tetrahydrofuranyl, tetrahydropyranyl, tetrahydropyrindinyl, tetrahydropyrimidinyl, tetrahydrothiophenyl, tetrahydrothiopyranyl, and the like. One way to substitute a heteroatom is to substitute a protected group. For example, a hydrogen on a nitrogen can be replaced with a tertbutoxycarbonyl. Optionally, the heterocyclyl can be substituted with one or several substituents. This definition only considers stable heterocyclic group isomers.
“Heteroaromatic?”, ‘heteroaryl? are terms used herein. A monocyclic or polycyclic heteroaromatic heteroaromatic rings comprising carbon atom-ring members and one to several heteroatom ring member. Each heteroatom can be selected independently from nitrogen, which can also be oxidized (e.g. N(O), quaternized, oxygen, and sulfur, including sulfoxide or sulfone). Representative heteroaryl groups include pyridyl, 1-oxo-pyridyl, furanyl, benzo[1,3]dioxolyl, benzo[1,4]dioxinyl, thienyl, pyrrolyl, oxazolyl, imidazolyl, thiazolyl, a isoxazolyl, quinolinyl, pyrazolyl, isothiazolyl, pyridazinyl, pyrimidinyl, pyrazinyl, a triazinyl, triazolyl, thiadiazolyl, isoquinolinyl, indazolyl, benzoxazolyl, benzofuryl, indolizinyl, imidazopyridyl, tetrazolyl, benzimidazolyl, benzothiazolyl, benzothiadiazolyl, benzoxadiazolyl, indolyl, tetrahydroindolyl, azaindolyl, imidazopyridyl, quinazolinyl, purinyl, pyrrolo[2,3]pyrimidinyl, pyrazolo[3,4]pyrimidinyl, imidazo[1,2-a]pyridyl, and benzothienyl. One embodiment of heteroaromatic rings is made up of 5-8 membered monocyclic heteroaryl ring. A heteroaromatic ring or heteroaryl ring can be attached at either a carbon or heteroatom of the heteroaromatic, heteroaryl rings. Optionally, heteroaryl groups can be substituted with one or several substituents.
“C5Heteroaryl” is the term used herein. An aromatic heterocyclic, 5-member aromatic heterocyclic, in which at least one carbon atom is replaced by a heteroatom, such as oxygen, sulfur, or nitrogen. Representative (C5)heteroaryls are furanyl and thienyl.
“As used herein the term?(C6)heteroaryl” An aromatic heterocyclic, 6-member aromatic heterocyclic, in which at least one carbon atom is replaced by a heteroatom, such as oxygen, nitrogen, or sulfur. Representative (C6)heteroaryls are pyridyl, pyridazinyl, pyrazinyl, triazinyl,etrazinyl and other similar compounds.
“Heteroaralkyl” is the term used herein. A heteroaryl group that has been attached to another group using a (C1?C6)alkylene. Representative heteroaralkyls include 2-(pyridin-4-yl)-propyl, 2-(thien-3-yl)-ethyl, imidazol-4-yl-methyl and the like. Optionally, heterooaralkyl groups can be substituted with one or several substituents.
“As used herein the term ‘halogen? “Halogen” or?halo is a term that means?F,?Cl,?Br or?I.” Meaning?F?,??Cl, or?Br?
“Heteroalkyl” is the term used herein. “Heteroalkyl” is a straight or branched chain-alkyl group in which one or more internal carbon atoms are replaced by a heteroatom. For example, O, N, or S. [CH2]y[CH3] in which x is a positive number and y is either 0 or positive integer. The replacement of the carbonatom does not create an unstable compound.
“Suitable substitutes for an alkyl or alkenyl and alkyl, cycloalkyls, cycloalkenyls, heterocyclyls, and heteroaryls include:?C(O)R33,?C(O)R33,?C(O)R33,??C(O]R33,?C (O/R33),???C(O]R33,???C?)R33,???C(S)NR28R29, aralkyls, cycloalkyls, aryls, cycloalkenyls, aryls, cycloalkyls, cycloalkenyls, cycloalkyls, cycloalkyls, aryls, cycloalkyls, aryls, aryls, aryls, aryl, aryl, aryl cycloalkenyl cycloalkyl cycloalkenyl cycloalkyl cycloalkenyl aryl, cycloalkyl cycloalkenyl aryl, aryl cycloalkenyl aryl, aryl cycloalkenyl cycloalkyl aryl, aryl aryl, heteroaryl aryl, aryl aryl,, aryl aryl,, aryl aryl,, aryl aryl,, aryl, aryl aryl aryl aryl aryl aryl aryl aryl aryl aryl ary, aryl aryl aryl aryl aryl aryl aryl aryl aryl cycloalkenyl aryl, aryl aryl, aryl aryl, aryl aryl, aryl aryl, heteroaryl aryl aryl aryl aryl aryl aryl aryl aryl aryls, aryl aryl aryl aryl ary “Suitable substituents for an Alkyl or alkylene, Alkenyl or alkyl or heterocyclyl include an optionly substituted or substituted an alkyl or alkenyl; likewise, R28 and R29 are independent of each other.
“In addition, alkyl and cycloalkyl and alkylene, as well as any saturated portion of an alkenyl or cycloalkenyl and alkynyl group, can be substituted for?O?S??R32.”
A heterocyclyl, heteroaryl or heteroaralkyl group may contain a nitrogen atom. It can be substituted or unsubstituted. A substitute is a nitrogen atom found in an aromatic ring of heteroaryl groups. This nitrogen could be a quaternary.
“As used herein the terms’subject?, and?patient? “Subject?,?patient??” and “mammal?” are interchangeable. They are interchangeable. The terms’subject? and?patient? are interchangeable. The terms?subject? and?patient are interchangeable. Refers to an animal, such as a bird like a chicken, turkey or turkey, or a mammal. Preferably a mammal that includes a non-primate (e.g. a cow, horse, pig or horse, rabbit, or guineapig), and a primate, (e.g. a monkey, human, or chimpanzee). One embodiment of the subject is a nonhuman animal, such as a horse, cow or pig, or a pet (e.g. a dog, cat or guineapig). Preferably, the subject is a person.
“The term ‘lower’ as used herein refers to a group that has up to four atoms. A group with up to four atoms refers to. A?lower-alkyl? is an example. A?lower-alkyl? refers to an alkyl radical with 1 to 4 carbonatoms. ?O? (C1-C4)alkyl, and a?lower alkenyl or?lower-alkynyl? Refers to alkenyl and alkynyl radicals having between 2 and 4 carbon atoms, depending on their type.
“Unless otherwise indicated, compounds of the invention that contain reactive functional groups (such (without limitation), carboxy, hydroxy and thiol) also include protected derivatives. ?Protected derivatives? Protected derivatives are compounds that have a reactive site (or sites) blocked by one or more protecting groups. Examples of suitable protecting groups for hydroxyl groups include benzyl, methoxymethyl, allyl, trimethylsilyl, tert-butyldimethylsilyl, acetate, and the like. Examples of suitable amine protecting groups include benzyloxycarbonyl, tert-butoxycarbonyl, tert-butyl, benzyl and fluorenylmethyloxy-carbonyl (Fmoc). The following are examples of suitable thiol-protecting groups: acetyl and methoxymethyl, tertbutyl, Acetyl, Tert-Butyl, tertyl, and the like. Others suitable protecting groups are also well-known to those with ordinary skill in art. These include those found in T. W. Greene, Protecting Groups in Organic Synthesis, John Wiley & Sons, Inc. 1981.”
“Compound(s),” as used in this invention, means: Similar terms are used to refer to a compound of formula I-VIII, or Table 1, or any pharmaceutically acceptable salts, solvate clathrates, hydrates, polymorph, or prodrug thereof. They also include protected derivatives.
The compounds of the invention can contain one or more double bonds or chiral centers. They are stereoisomers and may exist as enantiomers or diastereomers, enantiomers or geometric isomers. The chemical structures described herein, which include the compounds of this invention are all the corresponding compounds’ geometric isomers, enantiomers and diastereomers. This means that both the stereochemically pure form of the compound (e.g. geometrically pure, anenantiomerically pur or diastereomerically pur) and isomeric combinations (e.g. enantiomeric diastereomeric, and geometric isomeric mixed). Sometimes, one enantiomer and one diastereomer, or one of the geometric isomers, will have superior activity, toxicity, or kinetic profile than other isomers. These enantiomers/diastereomers and geometries of compounds of the invention are preferred in these cases.
“Polymorph” is the term used herein. The term “polymorph” refers to solid crystalline forms or complexes of the compound of the invention. Different polymorphs can have different physical, chemical, and/or spectroscopic characteristics. The physical properties of different polymorphs include stability (e.g. to heat or light), compressibility, density (important in formulations and product manufacturing) and dissolution rates (which may affect bioavailability). Changes in chemical reactivity, such as differential oxidation, can cause differences in stability. Or, they may result in changes in mechanical characteristics, such as tablets crumbling on storage because a kinetically preferred polymorph becomes a thermodynamically stable polymorph. Processing can be affected by different physical properties of polymorphs. One polymorph may be more likely than another to form solvates, or be more difficult to filter out or wash clean of impurities. This could be due to its shape or size distribution.
“The term “hydrate” is used herein. “hydrate” refers to a compound according to the invention, or a salt thereof. It also includes a stoichiometric (or non-stoichiometric) amount of water bound by noncovalent intermolecular forces.
“As used in this invention, the term clathrate” A compound of the invention or a salt thereof, in crystal lattice form, that has spaces (e.g. channels) that contain a guest molecule (e.g. water or solvent) within.
“Prodrug” is the term used herein, unless indicated otherwise. A derivative of a chemical compound that can be hydrolyzed, oxidized, or other react under biological conditions (in vivo or in vitro) to make a compound according to this invention. These prodrugs can become active under biological conditions or may still be active in unreacted form. This invention contemplates prodrugs that include analogs or derivatives from compounds of formula I-(VIII), and Table 1. These may include biohydrolyzable moieties like biohydrolyzable ester, biohydrolyzable carbamates or biohydrolyzable carbonates as well as biohydrolyzable analogues of phosphates. Prodrugs can also include derivatives or derivatives of compounds from formula (I-(VIII), Table 1, and compounds that contain?NO,?NO2,?ONO or?ONO2 moieties. You can prepare prodrugs using well-known methods such as those in 1 BURGER?S MEDICINAL CHEMISTRY AND DRUGS DISCOVERY (1995), 172-178, 949-982, Manfred E. Wolff’s 5th ed.
“As used herein, and unless otherwise noted, the terms ‘biohydrolyzable amino?, biohydrolyzable ester?”,??biohydrolyzable carbamate?”,???biohydrolyzable carbonate?”,???biohydrolyzable urine?. ?biohydrolyzable analogue of phosphate? An amide, ester or carbamate, carbonate or ureide is a compound that does not destroy its biological activity but confers on it advantageous properties such as increased water solubility, reduced metabolism (e.g. because of decreased metabolism of the prodrug), better uptake, longer duration of action or improved onset or end result; or 2) is biologically inactive, but is transformed in vivo into a biologically active compound. Examples of biohydrolyzable amides include, but are not limited to, lower alkyl amides, ?-amino acid amides, alkoxyacyl amides, and alkylaminoalkylcarbonyl amides. Biohydrolyzable esters can include lower alkyl esters and alkoxyacyloxy ester, as well as alkyl acylamino orkyl esters and choline esters. Biohydrolyzable carbamates can include substituted ethylenediamines and lower alkylamines as well as aminoacids and hydroxyalkylamines.
“As used herein, ?Hsp90? Each member of the heat shock protein family has a mass of approximately 90-kiloDaltons. The Hsp90 is a member of the Hsp90 family, which is highly conserved in humans. Hsp90 and Hsp90. These isoforms include GRP94 which can be found in endoplasmic retinal, and HSP75/TRAP1, which can be found in mitochondrial matrix.
“c-Met” is a receptor Tyrosine Kinase, which is found in both normal and malignant cells. It has been identified to be a proto-oncogene. The HGF/cMet signaling program triggers an invading growth program. Although it is essential for embryonic development, when not properly controlled, this can lead to malignant growth, motility and migration, as well as invasion. This mechanism is still being studied. The Met gene in humans is located on chromosome 7, band 7q21-q31 and covers more than 120kb (Ma et al. Cancer and Metastasis Reviews 2003, 22:309-325). Wild type cells have c-Met as a heterodimer. It consists of an extracellular subunit and a larger intracellular?subunit. Functional structures and domains for c-Met include 1) a Sema domain at N-terminus that includes a MRS-rich region; 2) PSIdomain which is also found within plexins, Semaphorins, and Integrins; 3) IPT Repeats which can be found in immunoglobulins, plexins, and transcription factors; 4) a transmembrane and 5) juxtamembrane; and 6) the intracellular Tyrosine kinae kinae kinae kinae kinae ae and tyrosine kinae ae and the C-terminus.
“Activation by c-Met signaling depends on phosphorylation at multiple residues of c.Met. HGF binding activates c-Met’s kinase activity by activating autophosphorylation at Y1230 and Y1234. HGF binding can trigger phosphorylation of Y1313. This is necessary for binding PI3K, which is involved in prosurvival signaling. The C-terinus at c-Met activates the multisubstrate sign transducer docking station, which mediates interactions between SHC, Src and Gab1. It also recruits Grb2, PI3K, PLC and? Only phosphorylation Y1356 is required for SHP2 and SHC2. The Y1365 receptor is responsible for controlling cell morphogenesis. The binding of c?Cbl is mediated by phosphorylation of the Y1003 residue within the juxtamembranedomain. cBl acts as a negative regulator protein for cMet by encouraging the polyubiquitinization cMet, which leads to its degradation.”
“Dysregulation in HGF/cMet signaling can occur by: 1) increased expression HGF; 2) activating mutants that typically occur in either the tyrosine-kinase or the juxtamembrane cMet domains and confer constitutive activity; 3) intrachromosomal amplification of the Met gene and overexpression of c?Met; 4) chromosomal transfer such as in Trp/Met fusion protein, which results in the loss and also cause the juxtamembranembranembranembranembranembranembranembranembranembranembra domain to be s to activated and also result in constitutive activation.
In hereditary and sporatic renal carcinoma, ovarian carcinoma, metastatic head-and neck squamous cells carcinomas, NSCLC and SCLC, glioma and breast cancer, activating mutations in the tyrosine domain or the juxtamembranedomain of c?Met have been found. Amino acid residues M1268 (e.g. M1248D, M1248H), M1246 (e.g. Y1246H), M1230 (e.g. Y1230C), Y1213 (e.g. L1213V), and H1124 (e.g. H1112L and H1112Y) have been identified as activating mutations in somatic papillary kidney cell carcinoma. In germline papillary kidney cell carcinoma, activating mutations were found at the amino acid residues M1248 (e.g. Y1246 (e.g. Y1246N), L1238 (e.g. V1238I), L1230C (e.g. Y1230C, and Y1230H), and V1206 (e.g. V1206L), H1124D, H1112L, and H1112Y), and V111262R), K1262 (e., K1262R), e. (e.) and H111246R), e., and e., Y1246R), Y1248C), M1248I), M1246C, e., and e., and Y1246, Y1248T, and Y1246, e., and e., and e., and e., and H111238I), and Y11121246, and e., and e., and e., and e. Hepatocellular carcinoma has activating mutations found at amino acids M1268 (e.g. M1268I), M1262 (e.g. K1262R), T1191 (e.g. T11910). Head and neck squamous cells carcinoma activating mutations were found at the amino acid residues M1268 (e.g.., M1268I), K1262 (e.g.., K1262R), and T1191 (e.g.., T1191). G1137V is the amino acid residue that activates mutations in glioma. NSCLC activating mutants were found at amino acids residue T1010 (e.g. T1010I). SCLC has activated mutations at amino acids residues R988 (e.g. R988C) or T1010 (e.g. T1010I). Activating mutations in breast cancer have been identified at amino acids residues T1010 (e.g. T1010I). On the other hand, activating mutations in gastric cancer have been identified at amino acid residue T1010 (e.g. P1010I). The amino acids listed herein for C-Met are numbered according to Schmit, et. al., Onogene (1999), 18, 2343-2350. The invention’s compounds cause the degradation of cMet. They can be used alone or with other anticancer therapies in order to treat patients with cancers with activating mutations in either the tyrosine kinase or juxtamembrane domains of c.Met.
“The juxtamembrane, a receptor tyrosine-kinase component, has been shown to suppress catalytic function. Mutations in the juxtamembrane can relieve this repression and lead to oncogenesis. The 5 is replaced by the Tpr/Met Fusion Protein. The Met gene is replaced with Tpr, which contains two strong dimerization motif. Dimerization activates Met kinase activity, which results in metastatic and transformative properties. Tpr/Met Fusion Protein has been discovered in gastric cancer. It results in an increase in Met kinase activation. A small-cell lung cancer has also revealed a different splicing method for Met mRNA that skips the juxtamembrane. Met kinase activity is increased and oncogenesis is enhanced by the loss of the juxtamembranedomain. The invention compounds cause the degradation c-Met. They can be used alone or with other anticancer therapies in combination to treat cancer patients who have juxtamembrane mutations and deletions in c.Met.
“Amplification of the Met gene, overexpression of c?Met, has been seen in many types of cancers, including breast cancer, esophageal carcinoma, small cell lung, non-small-cell lung cancer, gastric cancer, esophageal, esophageal, stomach, lung, and breast cancers, as well as colorectal metastases. The invention’s compounds cause the degradation c-Met. They can be used alone or with other anticancer therapies in order to treat patients with metastatic cancers.
“Met amplification, mutation has been suggested as a strategy to make certain cancers resistant to treatment (e.g. chemotherapy or radiation therapy). Certain non-small cell lung carcinomas have an activating mutation of receptor tyrosinekinase (EGFR) which causes oncogenesis. Most EGFR-mutant NSCLCs respond initially to EGFR inhibitors like Tarceva and Iressa, but most of these tumors eventually become resistant to the drug. A subset of these resistant cancers have been shown to have amplified Met, and it is thought that Met amplification is a mechanisms of acquired resistance, and in particular acquired resistance to kinase inhibitors such as Iressa and Tarceva (EGFR inhibitors), Gleevec (a Bcr-Abl, PDGF, and c-Kit inhibitor) (Engelman et al., Sciencexpress, www.sciencexpress.org/26 Apr. 2007/page 1/10.1126/science. 1141478). The invention’s compounds cause the degrading of c-Met. They can be used alone or in combination, such as with kinase inhibitors to treat cancer patients who have become resistant to anticancer treatments.
“C-Met associated cancer” is the term used herein. Any type of malignant growth, metastasis or other disease that is caused or exacerbated by dysregulation of HGF/cMet signaling.
“B-raf” is a serine/threonine kinase involved in the MAP kinase pathways. It is encoded by a gene on chromosome7q32. Ten B-raf variants have been identified, which were created by splicing variants. The term “B-raf” refers to all such splicing variants. All such variants of splicing are called?B-raf? Three conserved regions (CRs) are found in B-raf: 1) CR1 contains a cysteine rich (CRD), most of the Ras binding site (RBD), and facilitates B-raf binding to Ras and its recruitment to the cell membrane. 2) CR2 contains a rich in serine, threonine, and the 5365 residue, which is an inhibitory-phosphorylation site. 3) CR3 contains the kinasedomain, a GXGXG motif, and regulatory phosphorylation spots 54468, D449 and T599, D448, D448, D449 and D449, D449, D448, D449 and D449, D449, D448, T599, 562, D447, D447, D448, S447, D447, D447, D449, D449, D449, D449, D447, D448, D447, D447, D447, and the regulatory phosphorylation sites, D449, and the GXGXGXGXGXGXGXGXGXGXGXGXGXGXGXGX59, D449, and the B599, and the B-p539. B-raf can be translocated to cells membranes and activated through association with GTP-bound Ras. B-raf’s conformation changes regulate it and makes it inactive when its activation segment is kept in inactive form due to hydrophobic interactions. The activation segment is phosphorylated, resulting in a shift towards the active conformation. It is interesting that the majority of BRAF oncogenic mutations occur in the area where the activation section and P-loop interact. This suggests that B-raf mutations cause the inactive B raf conformation to be destabilized, thereby encouraging an active B raf. “Berram, et.al., Journal of Clinical Oncology 2005, 23(27),:6771-6790.
B-raf-associated cancers are those in which there is abnormal B-raf activity. B-raf-associated cancers are those that have higher B-raf activity than wild type B.raf. V600E,V600D, V596R, V594V. G469A. G469E. G466V. and G464V are all activating mutations in kinase. Malignant melanomas include anaplastic thyroid carcinomas, papillary cancers, ovarian cancer, para-follicular medullary C-cell thyroid cancer, Barrett’s esophageal cancer, and head and neck squamous cells carcinoma.
“As used herein, ?NPM-ALK? A fusion protein is a protein that results from a translocation of the NPM/B23 nucleolar proteins gene sequence into the sequence which encodes the tyrosinekinase ALK sequence. The NPM-ALK Fusion Protein typically contains the first 117 amino acid of NPM’s amine terminal and the C-terminal residues 1058-1620 of ALK. FIG. shows a schematic representation NPM-ALK. 1 of Duyster, and al., Oncogene (2001), 20,:5623-5637. The entire teachings are included herein by reference
“NPM-ALK-associated cancers” is the term. NPM-ALK-associated cancers are those in which the NPM fusion protein is expressed. Examples include ALCL and diffuse large-b-cell lymphomas.
“C-kit” is a term that refers to a specialized type of kinase. or ?c-kit kinase? The membrane receptor protein Tyrosine Kinase is activated by binding Stem Cell Factor to its extracellular domain (Yarden and al. 1987; Qiu and al. 1988). Yarden, et. al. 1987, EMBO Journal, 11:3341-3351. Qiu, et. al. 1988, EMBO Journal, 7:1003-1011. These are incorporated herein in their entirety. The term “c-kit” refers to mutant versions of ckit kinase. or ?c-kit kinase? These include those that are classified as either (1) having one amino acid substitution at codon816 of the human C-kit Kinase or an equivalent position in another species (Ma and al. 1999, J. Invest Dermatol. 112:165-170. (2) Those with mutations involving the putative juxtamembrane zhelix of this protein (Ma et al. 1999, J. Biol. Chem., 274:13399-13402). These publications, as well as any drawings, are incorporated herein by reference.
“As used herein, ?Bcr-Abl? is a fusion protein, which results from translocation of gene sequences from the c-ABL protein Tyrosine Kinase on chromosome 9, into the BCR sequences chromosome 22, creating the Philadelphia chromosome. FIG. 1 shows a schematic representation of human Abl, Bcr and Bcr. 1. of U.S. Patent Application Ser. No. No. 10/193,651, filed Jul. 9 2002, whose entire teachings are included herein by reference. Depending on the breaking point of the Bcr gene fusion proteins, they can range in size from 185-230 kDa to transforming activity. However, they must contain the OLI domain (from Bcr) and the TK domain (from Abl). P230 Bcr Abl, P210Bcr -Abl and P190Bcr -Abl are the most commonly found Bcr/Abl gene products in humans. P210 Bcr Abl is characteristically CML, while P190 Bcr -Abl can be used to identify ALL.
“FLT3 kinase” is a tyrosine-kinase receptor that regulates and stimulates cellular proliferation. (See Gilliland et.al., Blood (2002) 100:1532-42. The entire teachings are included herein by reference). The FLT3 kinase contains five immunoglobulin-like regions in its extracellular area as well as an insert domain of 75-100 amino acid in the middle of it cytoplasmic region. The FLT3 kinase activates upon binding to the FLT3ligand. This causes receptor dimerization. Dimerization of the FLT3 kinase by FLT3 ligand activates the intracellular kinase activity as well as a cascade of downstream substrates including Stat5, Ras, phosphatidylinositol-3-kinase (PI3K), PLC, Erk2, Akt, MAPK, SHC, SHP2, and SHIP (see Rosnet et al., Acta Haematol. (1996), 95:218; Hayakawa et al., Oncogene (2000), 19:624; Mizuki et al., Blood (2000), 96:3907; and Gilliand et al., Curr. Opin. Hematol. (2002), 9(1): 274-81. The entire teachings from each of these references are included herein by reference. Both membrane-bound FLT3 and soluble ligands bind, dimerize and then activate FLT3 kinase.
“Normal cells that express FLT3 Kinase include immature cells of the hematopoietic cell line, which are typically CD34+ cells and brain (see Rosnet, Rosnet, and Small, Blood (1993), 8:1110-19; Small, et.al., Proc. Natl. Acad. Sci. Sci. But, FLT3 kinase is not sufficient to stimulate proliferation. The critical role of FLT3 kinase in immune function is also played by its regulation of dendritic cells proliferation and differentiation (see McKenna, Blood (2000) 95:3489-97; the entire teachings are included herein by reference).
“Numerous hematologic malignancies are FLT3-expressing, with the most prominent being AML (see Yokota and al., Leukemia (1997) 11:1605-09; the entire teachings are included herein by reference). Other FLT3-expressing malignancies include myelodysplastic and T-cell acutely lymphoblastic lesions, B-precursor cells acute lymphoblastic lymphomas, and chronic myelogenous lesions (see Rasko, Leukemia (1995), 9, 2058-66; the entire teachings are included herein).
“Activating mutations are caused by FLT3 kinase mutations that are associated with hematologic malignancies. The FLT3 kinase activates constitutively without binding or dimerization of FLT3 Ligand. This stimulates cells to grow continually. There are two types of activating mutations: internal tandem duplications, (ITDs), and point mutations in the activating loop. The term “FLT3 kinase” is used herein. This term refers to both the wild type FLT3 Kinase as well as mutant FLT3 Kinases such activating mutations-prone FLT3kinases.
Summary for “Triazole compounds which modulate HSP90 activity”
Although great strides have been made in understanding the genetic abnormalities that lead to malignant cancer cells, the current chemotherapy is not satisfactory and the prognosis remains poor for most patients with this disease. The majority of chemotherapeutic agents target a specific molecular target that is thought to be responsible for the malignant phenotype. The proliferation of cells is controlled by a complex network signaling pathways. Most malignant cancers are caused by multiple genetic abnormalities within these pathways. It is unlikely that a therapy agent that targets one molecular target can cure cancer patients.
“Heat shock proteins (HSPs), are chaperone proteins that are upregulated in response to high temperatures and other environmental stressors, such as ultraviolet light and nutrient deprivation and oxygen deprivation. HSPs are chaperones for other cellular proteins (client proteins). They facilitate the proper folding and repair of client proteins and aid in the refolding misfolded client protein. There are many families of HSPs with their own sets of client proteins. Hsp90 is the most common HSP family, accounting for around 1-2% of proteins in cells that are not under stress but increasing to approximately 4-6% in cells under stress. Hsp90 inhibition results in the degradation of its client proteins through the ubiquitin protasome pathway. Hsp90’s client proteins are, unlike other chaperone proteins. They include protein kinases and transcription factors that aid in signal transduction. A number of Hsp90’s client proteins have been implicated in the development of cancer. Below are examples of Hsp90 client protein that have been implicated with the progression of cancer.
Her-2 is a transmembrane Tyrosine Kinase Cell Surface Growth Factor Receptor. It is found in normal epithelial cell. Her2 contains an extracellular domain that interacts and transmits extracellular growth signals to the cell’s nucleus. A high level of Her2 expression is associated with poor prognosis in many malignancies such as breast cancer and ovarian cancer.
“Akt kinase, a serine/threonine-kinase that is a downstream effector molecular of phosphoinositide 3kinase, is involved in protecting cells from apoptosis. Because it stimulates cell proliferation, Akt kinase may be involved in the development of cancer.
“Cdk4/cyclinD complexes are involved with phosphorylation retinoblastoma proteins which is an essential step in the progression of cells through the G1 phase. The half-life of newly synthesized Cdk4 is decreased by disrupting Hsp90 activity.
The Raf family of protooncogenes (A, B, and C-raf), was first discovered when C-raf was identified (raf-1). This was due to its homology in v-raf which is the transform gene of the mouse-sarcomavirus 3611. Later, A-raf, a transforming gene from the Mill Hill No. avaian retrovirus, was found by screening a cDNA database under low stringency conditions with a v?raf probe. B-raf, a homology to C-Rmil was also discovered. 2. The Ras/Raf/MEK/ERK pathway is mediated by the Raf protein family, also known as the?MAP kinase pathways. (MEK stands to?MAPK/ERK Kinase). ERK stands to?extracellularly regulated kinases? It has been linked to the development and progression of many cancers by upregulating cell division and proliferation. All raf proteins can activate the MAP kinase pathway. B-raf activates this pathway more effectively than A-raf and C-raf. Mutations in the gene that encodes B-raf are also more common in cancer. B-raf mutations were found in 60 to 70% of malignant melanomas and 35% to 69% in papillary thyroid carcinoma. They also occur in 35% to 16% of colon carcinomas, 35% to 69% in papillary thyroid caricinoma, 35% to 69% in low-grade ovarian cancer, 4% to 15% in colon cancer, 35% to 48% in head and neck squamous cells carcinoma, 35% to 6.8% in head and neck squamaromaf, cholangiocarcinomaroma, in breast cancer, and cholangiocarcinoma. The majority of B-raf mutations found in human cancers occur in the kinasedomain and cluster in exons 11, 15 and 16. These genes contain several regulatory phosphorylation site (S446, 5447 D448, D449 D449, D449, D449, S599 and S602). (Beeram, and colleagues, Journal of Clinical Oncology, 23(27), 6771-6790. T1799A is the most common mutation. It accounts for more that 80% of BRAF gene mutations and leads to a V600E mutation. V600E was previously known as V599E. The gene mutation was originally called T1796A. This was due to a mistake with the GenBank nucleotide sequence, NM 004333. The correct GenBank sequence is: NT 007914. It designates the protein mutant as V600E, and the gene mutation at T1799A. This will be the corrected numbering. This mutation mimics phosphorylation of B-raf’s activation segment. It inserts a negatively charged residue close to two activating sites for phosphorylation, T599, and 5602. This causes constitutively active B raf in an independent Ras-dependent manner. “Endocrine-Related Cancer (2005) 12:245-262.
The Hsp90 inhibitor 17AAG has been shown in cancer cells to stimulate B-raf’s degradation. Mutant forms of B raf are more susceptible to being degraded than the wild type. When 17AAG was applied to A375, a melanoma cell that has the V600E mutation, B-raf degraded faster than in CHL cells that contained wild-type B-raf. Other B-raf mutations, such as V600D,G469A and G469E, were found to degrade faster than wild type B.raf when they were transfected into COS cell lines. B-raf mutants E586K or L597V did not show any signs of degradation after cells were treated with 17AAG. It is believed that B-raf mutants E586K and L597V are clients of Hsp90. Mutated forms of B -raf, however, are more dependent upon Hsp90 for folding and stability. (Dias, et al., Cancer Res. (2005), 65(23): 10686-10691). Raf-1, a MAP 3-kinase(MAP3K), can activate the serine/threonine-specific protein kinases ERK1 or ERK2. Activated ERBs play an important part in controlling gene expression that is involved in cell division, cell differentiation, and cell migration.
Anaplastic large-cell Lymphoma (ALCL), a non-Hodgkin?s lymphoma, is characterised by the expression CD30/Ki-1. ALCL is normally caused by T-cells. However, some cases may have a null or B-cell phenotype. Diffuse large B-cell lymphomas are cases that arise from B cells. Around 60% of ALCL cases that express CD30/Ki-1 also have the chromosomal transfer t(2:5)(p23.q35), which fuses the nucleophosmin gene (NPM/B233) to the anaplastic lymphomakine (ALK) and produces the oncogenetic protein NPM-ALK with activity. ALK rearrangements were observed in specific types of ALCL: 1) 30% to 50% of pleomorphic ALCL; 2) more than 80% for monomorphic ALCL; 3) 75%?100% of small-cell cases; and 4) 60%?100% of lymphohistiocytic ALCL. NPM-ALK can transform fibroblasts, hematopoietic and primary bone marrow cells. It is believed to activate phosphatidylinositol-3 kinase (PI-3 kinase), which protects against apoptosis and stimulates mitosis via the RAS pathway. (Duyster, et al., Oncogene (2001), 20:5623-5637). NPM?ALK has been shown associate with Hsp90. Incubation of NPM?ALK-expressing ALCL cells with 17AAG of the benzoquinone isamycin has been shown not to disrupt this association. This can lead to increased degradation of NPM?ALK and cell cycle arrest. (Georgakis, et al., Exp. Hematology (2006), 34(12):1670-1679; Bonvini, et al., Cancer Research (2002), 62:1559-1566).”
“V-src is the Rous Sarcoma virus’s transforming protein. It is an example of an oncogene that induces cellular mutation (i.e. tumorogenesis) through non-regulated kinase activities. Hsp90 can interact with v-scr to inhibit its destruction.
“Hsp90 is necessary to maintain steroid hormone receptors at a conformation capable binding high affinity hormones with high affinity. The treatment of hormone-associated malignancies like breast cancer, such as Hsp90 inhibition, is expected to prove useful.
“P53 is a tumor suppressor protein which causes cell cycle arrest, and apoptosis. About half of all cancers are caused by mutations in the p53 gene. This makes it one of the most prevalent genetic alterations in cancerous cells. A poor prognosis is also associated with p53 mutation. Hsp90 interacts with wild-type P53, but mutant p53 forms a stronger association than wild-type Hsp53 due to its misfolded conformations. The mutated protein is protected from normal proteolytic degradation by a stronger interaction with Hsp90, which prolongs its half-life. A heterozygous cell for wild-type and mutated p53 will have mutant p53 degraded by inhibition of Hsp90’s stabilizing effect. This restores normal transcriptional activity for wild-type.
“Hif-1? Hypoxia-inducible transcription factors that are up-regulated in low oxygen conditions. Normal oxygen conditions, Hif-1 is degraded. Hif-1 is a tumor suppressor protein that associates with Von Hippel-Lindau? (VHL). This inhibits Hif-1’s ability to associate with VHL. Hif-1 to accumulate and form complexes with it? To form an active transcription complex which associates with hypoxia response elements to activate transcription of vascular epithelial growth factor. An increase in Hif-1? Increased Hif-1 is associated with poor prognosis and increased metastasis.”
There are two types of PKs. The protein tyrosine kinases, (PTKs), catalyze phosphorylation tyrosine kinase residues and the serine/threonine kinases kinases(STKs), catalyze phosphorylation threonine residues. The receptor tyrosine-kinases are growth factor receptors that have PTK activity. The family of receptor tyrosine-kinases is tightly controlled enzymes. Abnormal activation of different members of this family is a hallmark of cancer. There are subgroups within the receptor tyrosine-kinase family that share similar sequence and structural similarities.
“Epidermal growth factor receptor (EGFR), is a member the type 1 subgroup receptor tyrosine kinase families of growth factor receptors. These receptors play crucial roles in cellular differentiation, growth, and survival. These receptors are activated by specific ligand binding, which results in homo- or heterodimerization of receptor family members and subsequent autophosphorylation. EGFR is bindable to epidermal growth factors (EGF), transforming Growth Factor (?). (TGF?), amphiregulin, and other viral growth factors. Activating EGFR triggers a series of intracellular signaling pathways that are involved in both cellular proliferation (the ras/raf/MAP kinase pathway), and survival (the PI3 kinase/Akt pathway). Several members of this family, including HER2 and EGFR, have been implicated directly in cellular transformation.
“Many human malignancies are linked to aberrant or excessive expression of EGFR, and/or its specific ligands” (Gullick, Br. Med. Bull. (1991), 47.87-98; Modijtahedi, Int. J. Oncol. (1994), 4:277-96; Salomon, et al., Crit. Rev. Oncol. Hematol. (1995); 19;183-232. The entire teachings from each of these references are included herein by reference. Overexpression or aberrant EGFR has been linked to adverse prognosis for a variety of human cancers including head, neck, breast, colon and prostate (e.g. NSCLC, adenocarcinoma, squamous lung carcinoma, and squamous cell carcinoma), ovaries, gastrointestinal (gastric, colon and pancreatic), renal cancer, bladder cancer, glioma and gynecological cancers and prostate cancer. Overexpression of tumor EGFR can be associated with poor prognosis and chemoresistance in some cases (Lei et al. Anticancer Res. (1999), 19:221-8; Veale, et al., Br. J. J.
Gefitinib is a chemotherapeutic drug that inhibits EGFR activity. It has been shown to be very effective in a subset lung cancer patients with mutations in the tyrosine kinasedomain of EGFR. These mutants showed two to three times more activity when EGF was present than wild-type EGFR. The cells also absorbed wild type EGFR and it was down-regulated within 15 minutes. However, mutant EGFR continued to activate for as long as three hours. (Lynch, et. al., The New England Journal of Medicine (2006) 350:2129-2139; the complete teachings of which are incorporated by reference herein).
Another type of cancer is called gliomas. It is caused by amplification or mutation of the EGFR genes. A deletion of exons 2–7 results in a truncated EGFR form in which the amino acids 6-273 and a single glycine atom are replaced. This is one of the most common mutations of the EGFR genes. This mutation is known as EGFRvIII, and it is found in approximately half of all glioblastomas. EGFRvIII cannot bind TGF or EGF. It has constitutive, ligand independent tyrosinekinase activity. Hsp90 copurifies with EGFRvIII, indicating that Hsp90 interacts with EGFRvIII. Hsp90 inhibitor geldanamycin was able decrease the expression of EGFRvIII. This indicates that Hsp90 interacts with EGFRvIII. (Lavictoire, et.al., Journal of Biological Chemistry (2003) 278(7):5292-599; the complete teachings of which are incorporated in this reference). These results show that Hsp90 inhibition is an effective strategy to treat cancers associated with inappropriate EGFR activity.
“The type III group of receptor tyrosine kinases includes platelet-derived Growth Factor (PDGF) receptors, PDGF receptors alpha, beta, colony-stimulating factors (CSF-1R), c-Fms), Fms -like tyrosine kinase FLT3 and stem cell factor receptor c-kit. F1LT3 is expressed primarily on immature human hematopoietic stem cells and regulates their survival and proliferation.
Hematologic cancers are also called hematopoietic or hematologic malignancies. They include leukemia and lymphoma, as well as cancers of blood or bone marrow. Acute myelogenous (AML), a clonal hematopoietic, stem cell leukemia, is about 90% of acute leukemias in adults. It has an incidence rate of 3.9 per 100,000. (See Lowenberg et. al., N. Eng. J. Med. 341: 1051-62 (1999) and Lopesde Menezes, et al, Clin. Cancer Res. (2005), 11(14),:5281-5291. The enter teachings from both references are incorporated here by reference. Although chemotherapy can lead to complete remissions of AML, the long-term survival rate is only about 14%. There are approximately 7,400 AML related deaths each year in the United States. A majority of AML blasts are wild-type FLT3 with 25% to 35% expressing FLT3 kinase receiver mutations that result in constitutively active FLT. AML patients have two types of activating mutations: point mutations in the activating loop and internal tandem duplications. Patients with AML who have FLT3 mutations (ITDs) are indicative of poor survival rates. In patients in remission FLT3 mutations are the most detrimental factor in the relapse rate. 64% of patients with FLT3 mutations relapse within five years. Current Pharmaceutical Design (2005), 11/3449-3457. The entire teachings are included herein by reference. Clinical studies have shown that FLT3 mutations are predictive of a poor prognosis for AML patients. They may also be important in the development and maintenance the disease.
Mixed Lineage Leukemia (MLL), which involves translocations of chromosome 11, band q23 (11q23), occurs in approximately 80% of infants with hematological malignancies, and 10% in adult acute leukemias. Certain 11q23 translocations have been shown to be critical for immortalization of hematopoietic precursors in vitro. However, leukemia can only develop from a secondary genotoxic event. The strong correlation between FLT3 expression and MLL fusion gene gene expression is evident. FLT3 is the most frequently overexpressed gene of MLL. It has also been demonstrated that activating FLT3 and MLL fusion gene expression can induce acute leukemia, with a brief latency period (see Ono et al. J. of Clinical Investigation 2005, 115:919-929; the entire teachings of which have been incorporated herein). It is thought that FLT3 is involved in the maintenance and development of MLL (see Armstrong et al. Cancer Cell 2003, 3:73-183).
“The FLT3?ITD mutation can also be found in approximately 3% of adult myelodysplastic cases and some cases acute lymphocyticleukemia (ALL). (Current Pharmaceutical Design, 2005, 11:3449-3457).
“FLT3 was shown to be a client protein for Hsp90. 17AAG, a benzoquinone-ansamycin antibiotic that inhibits Hsp90 activation, has been shown not to interfere with the association of Flt3 and Hsp90. Treatment with 17?AAG was shown to inhibit the growth of leukemia cells that have either FLT3-ITD or wild-type FLT3 mutations. (Yao et al. Clinical Cancer Research 2003, 9:4483-4493; all teachings are included herein by reference).
“c-Kit, a membrane type III receptor protein Tyrosine Kinase that binds Stem Cell Factor to its extracellular region, is an example of a membrane-type III receptor protein tyrosinekinase. c-Kit is essential for normal hematopoiesis and tyrosinekinase activity. Mutations in c-kit may lead to ligand-independent Tyrosine Kinase Activity, Autophosphorylation and uncontrolled cell proliferation. A variety of pathological states have been linked to c-Kit activation and/or aberrant expression. Evidence for the contribution of cKit to neoplastic disease includes evidence of its association with leukemias, mast cell tumors, small-cell lung cancer, testicular carcinoma, and certain cancers of central nervous system and gastrointestinal tract. C-Kit has also been linked to neurofibromatosis and neuroectodermal sarcomas in the female genital tract. (Yang et al., J Clin Invest. (2003), 112:1851-1861; Viskochil, J Clin Invest. (2003), 112;1791-1793. The entire teachings from each of these references are included herein by reference. Hsp90 inhibitor 17AAG has been shown that c-Kit is a client protein for Hsp90. Kasumi-1 cells are an acute myeloidleukemia cell line with a mutation in the c-kit gene.
“c-Met” is a receptor Tyrosine Kinase, which is a client protein for Hsp90. It is encoded in the Met protooncogene. HGF (also known as scatter factor (SF), is the natural ligand for c-Met and causes a range of cellular responses including proliferation, survival and wound healing. It also triggers morphogenetic branching and morphogenesis. (Ma et.al., Cancer and Metastasis Reviews (2003) 22: 309-325. Although c-Met, and HGF can be found in many tissues, their expression is usually restricted to epithelial cells and mesenchymal cells, respectively. HGF and c-Met are essential for mammalian development. They have been shown to play an important role in cell migration, cell proliferation, survival, and organization 3D tubular structures (e.g. renal tubular cells, gland formation). In many human cancers, tumor growth, dissemination, and invasion can be caused by dysregulation of cMet and/or HGF. HGF and c-Met are high-expressed in many cancers, and their expression is associated with poor prognosis. (Christensen, and al., Cancer Research 2003, 63:7345-7355). c-Met receptor mutations were found in a variety of cancers, including ovarian cancer, childhood hepatocellular cancer, metastatic head & neck squamous cells carcinomas, esophageal carcinoma and gastric cancer. Met gene amplification, c-Met overexpression has been linked to non-small-cell lung cancer (NSCLC), small-cell lung cancer (SCLC), and colorectal carcinoma. The Tpr/Met fusion proteins has also been found in gastric cancer and osteogenic sarcoma. Multiple kidney tumors can be caused by germine mutations activating c-Metkinase. Many studies have shown that HGF and c-Met expression are linked to the progression of various types of cancer, including lung, colon and breast, prostate, liver and pancreas.
Gleevec’s success in targeting RTKs that are dysregulated by human cancers has been illustrated by its successes in targeting Bcr -Abl in chronic meelogenous Leukemia, c-Kit and gastroinstinal stromal tumours, Herceptin in Her-2-overexpressing breast cancers and Iressa in selected NSCLC with dysregulated EGFR. There is strong evidence to support the use of c-Met for treatment of human cancers. Several small drug molecules that inhibit c?Met are currently being developed. Therapies that target specific RTK can be effective in treating cancer, but they eventually fail because of additional mutations that allow RTK activity to continue with the drug. Moreover, SU11274, a selective cMet inhibitor, is highly effective against wild-type cMet as well as some mutants. However, it has not been proven to be effective against other cMet mutants (Berthou et al., Oncogene (2004) 23:5387-5393). It is therefore necessary to find new anticancer therapies that inhibit cMet’s expression or activity via a different mechanism from those that inhibit cMet directly.
“BCR-ABL, an ocoprotein that has tyrosinekinase activity, has been associated with chronic lymphocytic and acute leukemia. It also affects patients with acute myelogenous and acute lymphocytic lesions (ALL). The BCR-ABL cancer gene has been detected in approximately 2% of AML patients, 20 percent of ALL adults, 5% of ALL children, and at least 90-95% CML patients. Translocation of gene sequences from chromosome 9’s c-ABL protein tyrosine kinase into chromosome 22 results in the BCR-ABL, or Philadelphia chromosome. It has been demonstrated that the BCR-ABL gene can produce at least three alternative Chimeric Proteins, p230 Bcr -Abl p210 Bcr -Abl and p190 Bcr -Abl. These proteins have unregulated Tyrosine Kinase Activity. CML is associated most frequently with the p210BcrAbl fusion proteins, while ALL is associated more often with the p190Bcr-Abl. Bcr-Abl is also associated with various other hematological malignancies, including CML, lymphomas, myelomonocytic Leukemia, lymphomas, and erythroid Leukemia.
Studies have shown that BcrAbl activity and expression can be reduced to treat BcrAbl-positive lesions. Agents such as As2O3 that lower Bcr Abl expression have been proven to be very effective in fighting Bcr abl leukemias. Imatinib, also known as STI571 or Gleevic, inhibits Bcr Abl tyrosine kinase activation and induces differentiation and apoptosis. This causes the eradication in vivo and vitro of Bcr -Abl positive cells. Imatinib treatment is usually effective in inducing remission in patients with CML who are in the chronic phase as well as those in blast crises. In many cases, especially those who had been in blast crises before remission, the remission does not last because the Bcr/Abl fusion protein evolves mutations that make it resistant to Imatinib. ”
“Bcr fusion proteins are composed of Hsp90 complexes and are rapidly destroyed when Hsp90 action is blocked.” It has been demonstrated that geldanamycin is a benzoquinone-ansamycin antibiotic which disrupts the association between Bcr?Abl and Hsp90 results in proteasomal destruction of Bcr?Abl cells.
Mutational analysis has shown that Hsp90 is essential for normal eukaryotic cell survival. Hsp90 overexpression in cancer cells suggests that Hsp90 may play a critical role in cancer cell survival. Cancer cells might also be more sensitive than normal cells to Hsp90 inhibition. Cancer cells are known to have high levels of overexpressed and mutated oncoproteins, which depend on Hsp90 to fold. Hsp90 may also be important for tumor survival because of the hostile environment in which tumor cells are found. Hsp90 inhibition causes simultaneous inhibition of many oncoproteins as well as hormone receptors. This makes it a desirable target for anti-cancer agents. Clinical trials have shown that benzoquinone and ansamycins, which are natural products that inhibit Hsp90 in mice, can show therapeutic activity.
“Benzoquinone ansamycins and their derivatives are promising, but they have a few limitations. They are difficult to formulate because of their low oral bioavailability and limited solubility. They are also metabolized by polymorphic cytochrome CYP3A4 which makes them a substrate for the P-glycoprotein Export Pump, which is involved in multidrug resistance. There is a need for new therapies that improve the prognosis and overcome the limitations of current anti-cancer drugs.
HSPs are extremely conserved, from microorganisms all the way to mammals. Both the host and the pathogen increase HSP production when they infect a host. The infection process appears to have many roles for HSPs. Hsp90, for example, has been shown to be involved in the pathways that are responsible for the uptake and/or death of bacteria in phagocytic cell linings. Yan, L. et al., Eukaryotic Cell, 567-578, 3(3), 2004. Hsp90 is also essential for the uptake binary actin ADPribosylating toxins into eukaryotic cell. Haug. Infection and Immunity 12, 3066-3068. 2004. Hsp90 was also identified to play a role in viral proliferation in many viruses, including the influenza virus, vacciniavirus, herpes simplex virus types I and HIV-1 virus. Momose, F, et al., J. Biol. Chem., 45306-45314, 277(47), 2002; Hung, J., et al., J. Virology, 1379-1390, 76(3), 2002; Li, Y., et al., Antimicrobial Agents and Chemotherapy, 867-872, 48(3), 2004; O’Keefe, B., et al., J. Biol. Chem., 279-287, 275(1), 2000.”
Opportunistic fungal infections have been a growing problem, especially in immunocompromised patients. Hsp90 is implicated in the development of antifungal drug resistance in fungi. Cowen, L. et al., Eukaryotic Cell, 2184-2188, 5(12), 2006; Cowen, L. et al., Science, 309:2185-2189, 2005.”
“The present invention contains compounds that inhibit Hsp90 activity and can be used in the treatment proliferative diseases such as cancer.”
“In one embodiment, this invention provides compounds represented as structural formula (I).”
“wherein:”
“In one embodiment, of the compounds represented by formula (I), the compound is not 3-hydroxy-4-(5-mercapto-4-(naphthalen-1-yl)-4H-1,2,4-triazol-3-yl)phenyl dihydrogen phosphate.”
“Compounds shown in Table 1 and compounds of any formula therein, as well as tautomers or pharmaceutically acceptable salts of any formula, or compounds of any formula thereof, or solvates, chlorates, clathrates or hydrates, polymorphs, or prodrugs thereof, inhibit Hsp90’s activity and facilitate the degradation of Hsp90 clients proteins. Hsp90 is essential for normal eukaryotic cell survival. Hsp90 may be overexpressed in some tumor types, which suggests that it may play an important role in the survival and growth of cancer cells. Cancer cells might also be more sensitive than normal cells to Hsp90 inhibition. The compounds in Table 1, or any compounds of any formula, or tautomers or pharmaceutically acceptable salts, compounds, clathrates and hydrates, polymorphs, or prodrugs thereof are all useful for treating proliferative disorders like cancer.
Although chemotherapeutic drugs initially cause tumor regression in some cases, the majority of agents currently being used to treat cancer only target one pathway for tumor progression. In many cases, tumors that have been treated with multiple chemotherapeutic drugs develop multidrug resistance, and stop responding to treatment. Hsp90 inhibition has the advantage of inhibiting Hsp90’s activity. Many of its client proteins are protein kinases and transcription factors involved with signal transduction. This can lead to cancer progression. Inhibiting Hsp90 can be used to block multiple pathways that lead to tumor progression. The combination of Hsp90 inhibitors of the invention with other chemotherapy agents is more likely than any other therapies to cause tumor regression or elimination. It also makes it less likely that the tumor will develop multidrug resistance.
“A description of preferred embodiments is given below.”
“The present invention includes compounds described herein, and uses of those compounds for inhibiting Hsp90 activity as well as treatment of proliferative disorders such cancer. The invention includes compounds that can be used to stop or slow down the growth of cancerous cell or reduce or eliminate the number of such cells in a subject. Preferably, the subject is a mammal.
“In some embodiments, the compounds can be combined with other chemotherapy agents. This may reduce or prevent the development of multidrug resistant cancerous cell in mammal.” The compounds of this invention could allow for a lower amount of a second chemotherapy agent to be given to a mammal. This is because the invention compounds should prevent the development of multidrug resistant cells.
“The compounds of the invention may be used in certain embodiments to block, occlude or disrupt blood flow in the neovasculature.”
“In other embodiments, compounds of the invention may be used to treat or inhibit angiogenesis in a subject who is in need.”
“The invention also covers compounds that inhibit topoisomerase II activity.”
“The invention also relates the discovery that treating cells such as peripheral blood mononuclear cell (PMBCs), which have been stimulated by an inflammatory stimuli such as INF/LPS or SAC with an Hsp90 inhibit reduces expression of GR and the production of inflammatory cytokines.
“The invention also includes compounds that inhibit Hsp90’s activity and which are useful in treating or preventing infections.”
“In another embodiment, this invention pertains to a method for treating or preventing fungal drug resistant in mammal that is in need of such treatment.” This method involves administering an Hsp90 inhibitor to the mammal.
“In another embodiment, this invention concerns methods for administering a dosage solution containing compounds of the invention to a mammal.”
“A. TERMINOLOGY”
“Unless otherwise stated, the following terms are used herein:
“The term ‘alkyl’ as used herein means: “A saturated straight chain or branching non-cyclic hydrocarbon with between 1 and 10 carbon atoms. Representative saturated straight chain alkyls include methyl, ethyl, n-propyl, n-butyl, n-pentyl, n-hexyl, n-heptyl, n-octyl, n-nonyl and n-decyl; while saturated branched alkyls include isopropyl, sec-butyl, isobutyl, tert-butyl, isopentyl, 2-methylbutyl, 3-methylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2-methylhexyl, 3-methylhexyl, 4-methylhexyl, 5-methylhexyl, 2,3-dimethylbutyl, 2,3-dimethylpentyl, 2,4-dimethylpentyl, 2,3-dimethylhexyl, 2,4-dimethylhexyl, 2,5-dimethylhexyl, 2,2-dimethylpentyl, 2,2-dimethylhexyl, 3,3-dimtheylpentyl, 3,3-dimethylhexyl, 4,4-dimethylhexyl, 2-ethylpentyl, 3-ethylpentyl, 2-ethylhexyl, 3-ethylhexyl, 4-ethylhexyl, 2-methyl-2-ethylpentyl, 2-methyl-3-ethylpentyl, 2-methyl-4-ethylpentyl, 2-methyl-2-ethylhexyl, 2-methyl-3-ethylhexyl, 2-methyl-4-ethylhexyl, 2,2-diethylpentyl, 3,3-diethylhexyl, 2,2-diethylhexyl, 3,3-diethylhexyl and the like. The term “C1-C6alkyl” refers to a saturated straight chain alkyl. A saturated straight chain or branched, non-cyclic hydrocarbon with between 1 and 6 carbon atoms. The C1-C6 representative alkyl groups shown are those with between 1 and 6 carbon atoms. You can substitute any of the alkyl groups in compounds of this invention with one or more substituents.
“Alkenyl” is the term used herein. “A saturated straight chain, branched non-cyclic carbon having between 2 and 10 carbon atoms with at least one carbon carbon double bond. Representative straight chain and branched (C2-C10)alkenyls include vinyl, allyl, 1-butenyl, 2-butenyl, isobutylenyl, 1-pentenyl, 2-pentenyl, 3-methyl-1-butenyl, 2-methyl-2-butenyl, 2,3-dimethyl-2-butenyl, 1-hexenyl, 2-hexenyl, 3-hexenyl, 1-heptenyl, 2-heptenyl, 3-heptenyl, 1-octenyl, 2-octenyl, 3-octenyl, 1-nonenyl, 2-nonenyl, 3-nonenyl, 1-decenyl, 2-decenyl, 3-decenyl and the like. Optionally, alkenyl groups can be substituted with one or several substituents.
“Alkynyl” is the term used herein. “A saturated straight chain or branched, non-cyclic hydrocarbon with between 2 and 10 carbon atoms and at least one carbon-carbon triple bonds. The following are examples of straight chains and branched alkyls: 1-butyl; 2-butyl; 1-pentynyl; 2-pentynyl. 3-methyl-1-butynyl. 4-pentynyl. 1-octynyl. 2-octynyl. 7-octynyl. 1-nonyl. 2-decynyl. 8-nonyl. 1-decynynynynyl. Optionally, alkyl groups can be substituted with one or several substituents.
“Cycloalkyl” is the term used herein. “A saturated, mono- or multicyclic alkyl radical with between 3 and 20 carbon atoms. Representative cycloalkyls include cyclopropyl, 1-methylcyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl, -cyclodecyl, octahydro-pentalenyl, and the like. Optionally, cycloalkyl groups can be substituted with one or several substituents.
“As used herein the term “cycloalkenyl” means: “A mono- or poly-cyclic, non-aromatic alkyl-radical that has at least one carbon carbon double bond in the cycleic system and between 3 to 20 carbonatoms. Representative cycloalkenyls include cyclopentenyl, cyclopentadienyl, cyclohexenyl, cyclohexadienyl, cycloheptenyl, cycloheptadienyl, cycloheptatrienyl, cyclooctenyl, cyclooctadienyl, cyclooctatrienyl, cyclooctatetraenyl, cyclononenyl, cyclononadienyl, cyclodecenyl, cyclodecadienyl, 1,2,3,4,5,8-hexahydronaphthalenyl and the like. Optionally, cycloalkenyl groups can be substituted with one or several substituents.
“Haloalkyl” is the term used herein. An alkyl group in which one or more hydrogen radicals are replaced with a halo group. Each halo group is selected independently from?F?Cl??Br and?I. The term “halomethyl” is used. The term “halomethyl” refers to a methyl where one or more hydrogen radical(s), has been replaced by a Halo group. Examples of haloalkyl group include bromomethyl and trifluoromethyl.
“Alkoxy” is the term used herein. An alkyl group that is linked to another moiety by an oxygen linker.
“A ‘haloalkoxy’ is a term that means “as used herein.” An oxygen linker is a way to attach haloalkyl groups to other moiety.
“An?aromatic-ring” is the term used herein. Or?aryl? A hydrocarbon monocyclic, polycyclic, or polycyclic radical that has at least one aromatic ring. Examples of suitable aryl groups include, but are not limited to, phenyl, tolyl, anthracenyl, fluorenyl, indenyl, azulenyl, and naphthyl, as well as benzo-fused carbocyclic moieties such as 5,6,7,8-tetrahydronaphthyl. Optionally, aryl groups can be substituted with one or several substituents. One embodiment of the aryl group is monocyclic, and the ring contains 6 carbon atoms. This is called?(C6)aryl.
“Aralkyl” is the term used herein. An aryl group that has been attached to another group by a C1-C6 alkylene group. Aralkyl groups that are representative include naphth-3, 2-phenylmethyl, benzyl and other aryl groups. Optionally, aralkyl groups can be substituted with one or several substituents.
“The term ‘alkylene? as used herein is the following: Refers to an alkyl group with two points of attachment. The term “C1-C6alkylene” is used. An alkylene group with one to six carbon atoms refers to it. Prefer straight chain (C1-C6) alkylene groups. Methylene (?CH2?) is a non-limiting example of an alkylene group. ), ethylene (?CH2CH2? ), n-propylene (?CH2CH2CH2? ), n-propylene (?CH2CH2CH2CH2? ), and other similar. Optionally, Alkylene groups can be substituted with one or several substituents.”
“Heterocyclyl” is the term used herein. “Heterocyclyl” can be used to refer to a monocyclic (typically with 3- to 10 members) or polycyclic (7- to 20-members). It is either a saturated or unsaturated non-aromatic rings. A heterocycle with 3 to 10 members can have up to 5 heteroatoms, while a heterocycle with 7 to 20 members can have up to 7 heteroatoms. A heterocycle typically has at least one carbon-atom ring member. Each heteroatom can be selected independently from nitrogen. This can be oxidized (e.g. N(O), quaternized, oxygen, and sulfur. Any heteroatom or carbon atom can attach the heterocycle. Representative heterocycles include morpholinyl, thiomorpholinyl, pyrrolidinonyl, pyrrolidinyl, piperidinyl, piperazinyl, hydantoinyl, valerolactamyl, oxiranyl, oxetanyl, tetrahydrofuranyl, tetrahydropyranyl, tetrahydropyrindinyl, tetrahydropyrimidinyl, tetrahydrothiophenyl, tetrahydrothiopyranyl, and the like. One way to substitute a heteroatom is to substitute a protected group. For example, a hydrogen on a nitrogen can be replaced with a tertbutoxycarbonyl. Optionally, the heterocyclyl can be substituted with one or several substituents. This definition only considers stable heterocyclic group isomers.
“Heteroaromatic?”, ‘heteroaryl? are terms used herein. A monocyclic or polycyclic heteroaromatic heteroaromatic rings comprising carbon atom-ring members and one to several heteroatom ring member. Each heteroatom can be selected independently from nitrogen, which can also be oxidized (e.g. N(O), quaternized, oxygen, and sulfur, including sulfoxide or sulfone). Representative heteroaryl groups include pyridyl, 1-oxo-pyridyl, furanyl, benzo[1,3]dioxolyl, benzo[1,4]dioxinyl, thienyl, pyrrolyl, oxazolyl, imidazolyl, thiazolyl, a isoxazolyl, quinolinyl, pyrazolyl, isothiazolyl, pyridazinyl, pyrimidinyl, pyrazinyl, a triazinyl, triazolyl, thiadiazolyl, isoquinolinyl, indazolyl, benzoxazolyl, benzofuryl, indolizinyl, imidazopyridyl, tetrazolyl, benzimidazolyl, benzothiazolyl, benzothiadiazolyl, benzoxadiazolyl, indolyl, tetrahydroindolyl, azaindolyl, imidazopyridyl, quinazolinyl, purinyl, pyrrolo[2,3]pyrimidinyl, pyrazolo[3,4]pyrimidinyl, imidazo[1,2-a]pyridyl, and benzothienyl. One embodiment of heteroaromatic rings is made up of 5-8 membered monocyclic heteroaryl ring. A heteroaromatic ring or heteroaryl ring can be attached at either a carbon or heteroatom of the heteroaromatic, heteroaryl rings. Optionally, heteroaryl groups can be substituted with one or several substituents.
“C5Heteroaryl” is the term used herein. An aromatic heterocyclic, 5-member aromatic heterocyclic, in which at least one carbon atom is replaced by a heteroatom, such as oxygen, sulfur, or nitrogen. Representative (C5)heteroaryls are furanyl and thienyl.
“As used herein the term?(C6)heteroaryl” An aromatic heterocyclic, 6-member aromatic heterocyclic, in which at least one carbon atom is replaced by a heteroatom, such as oxygen, nitrogen, or sulfur. Representative (C6)heteroaryls are pyridyl, pyridazinyl, pyrazinyl, triazinyl,etrazinyl and other similar compounds.
“Heteroaralkyl” is the term used herein. A heteroaryl group that has been attached to another group using a (C1?C6)alkylene. Representative heteroaralkyls include 2-(pyridin-4-yl)-propyl, 2-(thien-3-yl)-ethyl, imidazol-4-yl-methyl and the like. Optionally, heterooaralkyl groups can be substituted with one or several substituents.
“As used herein the term ‘halogen? “Halogen” or?halo is a term that means?F,?Cl,?Br or?I.” Meaning?F?,??Cl, or?Br?
“Heteroalkyl” is the term used herein. “Heteroalkyl” is a straight or branched chain-alkyl group in which one or more internal carbon atoms are replaced by a heteroatom. For example, O, N, or S. [CH2]y[CH3] in which x is a positive number and y is either 0 or positive integer. The replacement of the carbonatom does not create an unstable compound.
“Suitable substitutes for an alkyl or alkenyl and alkyl, cycloalkyls, cycloalkenyls, heterocyclyls, and heteroaryls include:?C(O)R33,?C(O)R33,?C(O)R33,??C(O]R33,?C (O/R33),???C(O]R33,???C?)R33,???C(S)NR28R29, aralkyls, cycloalkyls, aryls, cycloalkenyls, aryls, cycloalkyls, cycloalkenyls, cycloalkyls, cycloalkyls, aryls, cycloalkyls, aryls, aryls, aryls, aryl, aryl, aryl cycloalkenyl cycloalkyl cycloalkenyl cycloalkyl cycloalkenyl aryl, cycloalkyl cycloalkenyl aryl, aryl cycloalkenyl aryl, aryl cycloalkenyl cycloalkyl aryl, aryl aryl, heteroaryl aryl, aryl aryl,, aryl aryl,, aryl aryl,, aryl aryl,, aryl, aryl aryl aryl aryl aryl aryl aryl aryl aryl aryl ary, aryl aryl aryl aryl aryl aryl aryl aryl aryl cycloalkenyl aryl, aryl aryl, aryl aryl, aryl aryl, aryl aryl, heteroaryl aryl aryl aryl aryl aryl aryl aryl aryl aryls, aryl aryl aryl aryl ary “Suitable substituents for an Alkyl or alkylene, Alkenyl or alkyl or heterocyclyl include an optionly substituted or substituted an alkyl or alkenyl; likewise, R28 and R29 are independent of each other.
“In addition, alkyl and cycloalkyl and alkylene, as well as any saturated portion of an alkenyl or cycloalkenyl and alkynyl group, can be substituted for?O?S??R32.”
A heterocyclyl, heteroaryl or heteroaralkyl group may contain a nitrogen atom. It can be substituted or unsubstituted. A substitute is a nitrogen atom found in an aromatic ring of heteroaryl groups. This nitrogen could be a quaternary.
“As used herein the terms’subject?, and?patient? “Subject?,?patient??” and “mammal?” are interchangeable. They are interchangeable. The terms’subject? and?patient? are interchangeable. The terms?subject? and?patient are interchangeable. Refers to an animal, such as a bird like a chicken, turkey or turkey, or a mammal. Preferably a mammal that includes a non-primate (e.g. a cow, horse, pig or horse, rabbit, or guineapig), and a primate, (e.g. a monkey, human, or chimpanzee). One embodiment of the subject is a nonhuman animal, such as a horse, cow or pig, or a pet (e.g. a dog, cat or guineapig). Preferably, the subject is a person.
“The term ‘lower’ as used herein refers to a group that has up to four atoms. A group with up to four atoms refers to. A?lower-alkyl? is an example. A?lower-alkyl? refers to an alkyl radical with 1 to 4 carbonatoms. ?O? (C1-C4)alkyl, and a?lower alkenyl or?lower-alkynyl? Refers to alkenyl and alkynyl radicals having between 2 and 4 carbon atoms, depending on their type.
“Unless otherwise indicated, compounds of the invention that contain reactive functional groups (such (without limitation), carboxy, hydroxy and thiol) also include protected derivatives. ?Protected derivatives? Protected derivatives are compounds that have a reactive site (or sites) blocked by one or more protecting groups. Examples of suitable protecting groups for hydroxyl groups include benzyl, methoxymethyl, allyl, trimethylsilyl, tert-butyldimethylsilyl, acetate, and the like. Examples of suitable amine protecting groups include benzyloxycarbonyl, tert-butoxycarbonyl, tert-butyl, benzyl and fluorenylmethyloxy-carbonyl (Fmoc). The following are examples of suitable thiol-protecting groups: acetyl and methoxymethyl, tertbutyl, Acetyl, Tert-Butyl, tertyl, and the like. Others suitable protecting groups are also well-known to those with ordinary skill in art. These include those found in T. W. Greene, Protecting Groups in Organic Synthesis, John Wiley & Sons, Inc. 1981.”
“Compound(s),” as used in this invention, means: Similar terms are used to refer to a compound of formula I-VIII, or Table 1, or any pharmaceutically acceptable salts, solvate clathrates, hydrates, polymorph, or prodrug thereof. They also include protected derivatives.
The compounds of the invention can contain one or more double bonds or chiral centers. They are stereoisomers and may exist as enantiomers or diastereomers, enantiomers or geometric isomers. The chemical structures described herein, which include the compounds of this invention are all the corresponding compounds’ geometric isomers, enantiomers and diastereomers. This means that both the stereochemically pure form of the compound (e.g. geometrically pure, anenantiomerically pur or diastereomerically pur) and isomeric combinations (e.g. enantiomeric diastereomeric, and geometric isomeric mixed). Sometimes, one enantiomer and one diastereomer, or one of the geometric isomers, will have superior activity, toxicity, or kinetic profile than other isomers. These enantiomers/diastereomers and geometries of compounds of the invention are preferred in these cases.
“Polymorph” is the term used herein. The term “polymorph” refers to solid crystalline forms or complexes of the compound of the invention. Different polymorphs can have different physical, chemical, and/or spectroscopic characteristics. The physical properties of different polymorphs include stability (e.g. to heat or light), compressibility, density (important in formulations and product manufacturing) and dissolution rates (which may affect bioavailability). Changes in chemical reactivity, such as differential oxidation, can cause differences in stability. Or, they may result in changes in mechanical characteristics, such as tablets crumbling on storage because a kinetically preferred polymorph becomes a thermodynamically stable polymorph. Processing can be affected by different physical properties of polymorphs. One polymorph may be more likely than another to form solvates, or be more difficult to filter out or wash clean of impurities. This could be due to its shape or size distribution.
“The term “hydrate” is used herein. “hydrate” refers to a compound according to the invention, or a salt thereof. It also includes a stoichiometric (or non-stoichiometric) amount of water bound by noncovalent intermolecular forces.
“As used in this invention, the term clathrate” A compound of the invention or a salt thereof, in crystal lattice form, that has spaces (e.g. channels) that contain a guest molecule (e.g. water or solvent) within.
“Prodrug” is the term used herein, unless indicated otherwise. A derivative of a chemical compound that can be hydrolyzed, oxidized, or other react under biological conditions (in vivo or in vitro) to make a compound according to this invention. These prodrugs can become active under biological conditions or may still be active in unreacted form. This invention contemplates prodrugs that include analogs or derivatives from compounds of formula I-(VIII), and Table 1. These may include biohydrolyzable moieties like biohydrolyzable ester, biohydrolyzable carbamates or biohydrolyzable carbonates as well as biohydrolyzable analogues of phosphates. Prodrugs can also include derivatives or derivatives of compounds from formula (I-(VIII), Table 1, and compounds that contain?NO,?NO2,?ONO or?ONO2 moieties. You can prepare prodrugs using well-known methods such as those in 1 BURGER?S MEDICINAL CHEMISTRY AND DRUGS DISCOVERY (1995), 172-178, 949-982, Manfred E. Wolff’s 5th ed.
“As used herein, and unless otherwise noted, the terms ‘biohydrolyzable amino?, biohydrolyzable ester?”,??biohydrolyzable carbamate?”,???biohydrolyzable carbonate?”,???biohydrolyzable urine?. ?biohydrolyzable analogue of phosphate? An amide, ester or carbamate, carbonate or ureide is a compound that does not destroy its biological activity but confers on it advantageous properties such as increased water solubility, reduced metabolism (e.g. because of decreased metabolism of the prodrug), better uptake, longer duration of action or improved onset or end result; or 2) is biologically inactive, but is transformed in vivo into a biologically active compound. Examples of biohydrolyzable amides include, but are not limited to, lower alkyl amides, ?-amino acid amides, alkoxyacyl amides, and alkylaminoalkylcarbonyl amides. Biohydrolyzable esters can include lower alkyl esters and alkoxyacyloxy ester, as well as alkyl acylamino orkyl esters and choline esters. Biohydrolyzable carbamates can include substituted ethylenediamines and lower alkylamines as well as aminoacids and hydroxyalkylamines.
“As used herein, ?Hsp90? Each member of the heat shock protein family has a mass of approximately 90-kiloDaltons. The Hsp90 is a member of the Hsp90 family, which is highly conserved in humans. Hsp90 and Hsp90. These isoforms include GRP94 which can be found in endoplasmic retinal, and HSP75/TRAP1, which can be found in mitochondrial matrix.
“c-Met” is a receptor Tyrosine Kinase, which is found in both normal and malignant cells. It has been identified to be a proto-oncogene. The HGF/cMet signaling program triggers an invading growth program. Although it is essential for embryonic development, when not properly controlled, this can lead to malignant growth, motility and migration, as well as invasion. This mechanism is still being studied. The Met gene in humans is located on chromosome 7, band 7q21-q31 and covers more than 120kb (Ma et al. Cancer and Metastasis Reviews 2003, 22:309-325). Wild type cells have c-Met as a heterodimer. It consists of an extracellular subunit and a larger intracellular?subunit. Functional structures and domains for c-Met include 1) a Sema domain at N-terminus that includes a MRS-rich region; 2) PSIdomain which is also found within plexins, Semaphorins, and Integrins; 3) IPT Repeats which can be found in immunoglobulins, plexins, and transcription factors; 4) a transmembrane and 5) juxtamembrane; and 6) the intracellular Tyrosine kinae kinae kinae kinae kinae ae and tyrosine kinae ae and the C-terminus.
“Activation by c-Met signaling depends on phosphorylation at multiple residues of c.Met. HGF binding activates c-Met’s kinase activity by activating autophosphorylation at Y1230 and Y1234. HGF binding can trigger phosphorylation of Y1313. This is necessary for binding PI3K, which is involved in prosurvival signaling. The C-terinus at c-Met activates the multisubstrate sign transducer docking station, which mediates interactions between SHC, Src and Gab1. It also recruits Grb2, PI3K, PLC and? Only phosphorylation Y1356 is required for SHP2 and SHC2. The Y1365 receptor is responsible for controlling cell morphogenesis. The binding of c?Cbl is mediated by phosphorylation of the Y1003 residue within the juxtamembranedomain. cBl acts as a negative regulator protein for cMet by encouraging the polyubiquitinization cMet, which leads to its degradation.”
“Dysregulation in HGF/cMet signaling can occur by: 1) increased expression HGF; 2) activating mutants that typically occur in either the tyrosine-kinase or the juxtamembrane cMet domains and confer constitutive activity; 3) intrachromosomal amplification of the Met gene and overexpression of c?Met; 4) chromosomal transfer such as in Trp/Met fusion protein, which results in the loss and also cause the juxtamembranembranembranembranembranembranembranembranembranembranembra domain to be s to activated and also result in constitutive activation.
In hereditary and sporatic renal carcinoma, ovarian carcinoma, metastatic head-and neck squamous cells carcinomas, NSCLC and SCLC, glioma and breast cancer, activating mutations in the tyrosine domain or the juxtamembranedomain of c?Met have been found. Amino acid residues M1268 (e.g. M1248D, M1248H), M1246 (e.g. Y1246H), M1230 (e.g. Y1230C), Y1213 (e.g. L1213V), and H1124 (e.g. H1112L and H1112Y) have been identified as activating mutations in somatic papillary kidney cell carcinoma. In germline papillary kidney cell carcinoma, activating mutations were found at the amino acid residues M1248 (e.g. Y1246 (e.g. Y1246N), L1238 (e.g. V1238I), L1230C (e.g. Y1230C, and Y1230H), and V1206 (e.g. V1206L), H1124D, H1112L, and H1112Y), and V111262R), K1262 (e., K1262R), e. (e.) and H111246R), e., and e., Y1246R), Y1248C), M1248I), M1246C, e., and e., and Y1246, Y1248T, and Y1246, e., and e., and e., and e., and H111238I), and Y11121246, and e., and e., and e., and e. Hepatocellular carcinoma has activating mutations found at amino acids M1268 (e.g. M1268I), M1262 (e.g. K1262R), T1191 (e.g. T11910). Head and neck squamous cells carcinoma activating mutations were found at the amino acid residues M1268 (e.g.., M1268I), K1262 (e.g.., K1262R), and T1191 (e.g.., T1191). G1137V is the amino acid residue that activates mutations in glioma. NSCLC activating mutants were found at amino acids residue T1010 (e.g. T1010I). SCLC has activated mutations at amino acids residues R988 (e.g. R988C) or T1010 (e.g. T1010I). Activating mutations in breast cancer have been identified at amino acids residues T1010 (e.g. T1010I). On the other hand, activating mutations in gastric cancer have been identified at amino acid residue T1010 (e.g. P1010I). The amino acids listed herein for C-Met are numbered according to Schmit, et. al., Onogene (1999), 18, 2343-2350. The invention’s compounds cause the degradation of cMet. They can be used alone or with other anticancer therapies in order to treat patients with cancers with activating mutations in either the tyrosine kinase or juxtamembrane domains of c.Met.
“The juxtamembrane, a receptor tyrosine-kinase component, has been shown to suppress catalytic function. Mutations in the juxtamembrane can relieve this repression and lead to oncogenesis. The 5 is replaced by the Tpr/Met Fusion Protein. The Met gene is replaced with Tpr, which contains two strong dimerization motif. Dimerization activates Met kinase activity, which results in metastatic and transformative properties. Tpr/Met Fusion Protein has been discovered in gastric cancer. It results in an increase in Met kinase activation. A small-cell lung cancer has also revealed a different splicing method for Met mRNA that skips the juxtamembrane. Met kinase activity is increased and oncogenesis is enhanced by the loss of the juxtamembranedomain. The invention compounds cause the degradation c-Met. They can be used alone or with other anticancer therapies in combination to treat cancer patients who have juxtamembrane mutations and deletions in c.Met.
“Amplification of the Met gene, overexpression of c?Met, has been seen in many types of cancers, including breast cancer, esophageal carcinoma, small cell lung, non-small-cell lung cancer, gastric cancer, esophageal, esophageal, stomach, lung, and breast cancers, as well as colorectal metastases. The invention’s compounds cause the degradation c-Met. They can be used alone or with other anticancer therapies in order to treat patients with metastatic cancers.
“Met amplification, mutation has been suggested as a strategy to make certain cancers resistant to treatment (e.g. chemotherapy or radiation therapy). Certain non-small cell lung carcinomas have an activating mutation of receptor tyrosinekinase (EGFR) which causes oncogenesis. Most EGFR-mutant NSCLCs respond initially to EGFR inhibitors like Tarceva and Iressa, but most of these tumors eventually become resistant to the drug. A subset of these resistant cancers have been shown to have amplified Met, and it is thought that Met amplification is a mechanisms of acquired resistance, and in particular acquired resistance to kinase inhibitors such as Iressa and Tarceva (EGFR inhibitors), Gleevec (a Bcr-Abl, PDGF, and c-Kit inhibitor) (Engelman et al., Sciencexpress, www.sciencexpress.org/26 Apr. 2007/page 1/10.1126/science. 1141478). The invention’s compounds cause the degrading of c-Met. They can be used alone or in combination, such as with kinase inhibitors to treat cancer patients who have become resistant to anticancer treatments.
“C-Met associated cancer” is the term used herein. Any type of malignant growth, metastasis or other disease that is caused or exacerbated by dysregulation of HGF/cMet signaling.
“B-raf” is a serine/threonine kinase involved in the MAP kinase pathways. It is encoded by a gene on chromosome7q32. Ten B-raf variants have been identified, which were created by splicing variants. The term “B-raf” refers to all such splicing variants. All such variants of splicing are called?B-raf? Three conserved regions (CRs) are found in B-raf: 1) CR1 contains a cysteine rich (CRD), most of the Ras binding site (RBD), and facilitates B-raf binding to Ras and its recruitment to the cell membrane. 2) CR2 contains a rich in serine, threonine, and the 5365 residue, which is an inhibitory-phosphorylation site. 3) CR3 contains the kinasedomain, a GXGXG motif, and regulatory phosphorylation spots 54468, D449 and T599, D448, D448, D449 and D449, D449, D448, D449 and D449, D449, D448, T599, 562, D447, D447, D448, S447, D447, D447, D449, D449, D449, D449, D447, D448, D447, D447, D447, and the regulatory phosphorylation sites, D449, and the GXGXGXGXGXGXGXGXGXGXGXGXGXGXGXGX59, D449, and the B599, and the B-p539. B-raf can be translocated to cells membranes and activated through association with GTP-bound Ras. B-raf’s conformation changes regulate it and makes it inactive when its activation segment is kept in inactive form due to hydrophobic interactions. The activation segment is phosphorylated, resulting in a shift towards the active conformation. It is interesting that the majority of BRAF oncogenic mutations occur in the area where the activation section and P-loop interact. This suggests that B-raf mutations cause the inactive B raf conformation to be destabilized, thereby encouraging an active B raf. “Berram, et.al., Journal of Clinical Oncology 2005, 23(27),:6771-6790.
B-raf-associated cancers are those in which there is abnormal B-raf activity. B-raf-associated cancers are those that have higher B-raf activity than wild type B.raf. V600E,V600D, V596R, V594V. G469A. G469E. G466V. and G464V are all activating mutations in kinase. Malignant melanomas include anaplastic thyroid carcinomas, papillary cancers, ovarian cancer, para-follicular medullary C-cell thyroid cancer, Barrett’s esophageal cancer, and head and neck squamous cells carcinoma.
“As used herein, ?NPM-ALK? A fusion protein is a protein that results from a translocation of the NPM/B23 nucleolar proteins gene sequence into the sequence which encodes the tyrosinekinase ALK sequence. The NPM-ALK Fusion Protein typically contains the first 117 amino acid of NPM’s amine terminal and the C-terminal residues 1058-1620 of ALK. FIG. shows a schematic representation NPM-ALK. 1 of Duyster, and al., Oncogene (2001), 20,:5623-5637. The entire teachings are included herein by reference
“NPM-ALK-associated cancers” is the term. NPM-ALK-associated cancers are those in which the NPM fusion protein is expressed. Examples include ALCL and diffuse large-b-cell lymphomas.
“C-kit” is a term that refers to a specialized type of kinase. or ?c-kit kinase? The membrane receptor protein Tyrosine Kinase is activated by binding Stem Cell Factor to its extracellular domain (Yarden and al. 1987; Qiu and al. 1988). Yarden, et. al. 1987, EMBO Journal, 11:3341-3351. Qiu, et. al. 1988, EMBO Journal, 7:1003-1011. These are incorporated herein in their entirety. The term “c-kit” refers to mutant versions of ckit kinase. or ?c-kit kinase? These include those that are classified as either (1) having one amino acid substitution at codon816 of the human C-kit Kinase or an equivalent position in another species (Ma and al. 1999, J. Invest Dermatol. 112:165-170. (2) Those with mutations involving the putative juxtamembrane zhelix of this protein (Ma et al. 1999, J. Biol. Chem., 274:13399-13402). These publications, as well as any drawings, are incorporated herein by reference.
“As used herein, ?Bcr-Abl? is a fusion protein, which results from translocation of gene sequences from the c-ABL protein Tyrosine Kinase on chromosome 9, into the BCR sequences chromosome 22, creating the Philadelphia chromosome. FIG. 1 shows a schematic representation of human Abl, Bcr and Bcr. 1. of U.S. Patent Application Ser. No. No. 10/193,651, filed Jul. 9 2002, whose entire teachings are included herein by reference. Depending on the breaking point of the Bcr gene fusion proteins, they can range in size from 185-230 kDa to transforming activity. However, they must contain the OLI domain (from Bcr) and the TK domain (from Abl). P230 Bcr Abl, P210Bcr -Abl and P190Bcr -Abl are the most commonly found Bcr/Abl gene products in humans. P210 Bcr Abl is characteristically CML, while P190 Bcr -Abl can be used to identify ALL.
“FLT3 kinase” is a tyrosine-kinase receptor that regulates and stimulates cellular proliferation. (See Gilliland et.al., Blood (2002) 100:1532-42. The entire teachings are included herein by reference). The FLT3 kinase contains five immunoglobulin-like regions in its extracellular area as well as an insert domain of 75-100 amino acid in the middle of it cytoplasmic region. The FLT3 kinase activates upon binding to the FLT3ligand. This causes receptor dimerization. Dimerization of the FLT3 kinase by FLT3 ligand activates the intracellular kinase activity as well as a cascade of downstream substrates including Stat5, Ras, phosphatidylinositol-3-kinase (PI3K), PLC, Erk2, Akt, MAPK, SHC, SHP2, and SHIP (see Rosnet et al., Acta Haematol. (1996), 95:218; Hayakawa et al., Oncogene (2000), 19:624; Mizuki et al., Blood (2000), 96:3907; and Gilliand et al., Curr. Opin. Hematol. (2002), 9(1): 274-81. The entire teachings from each of these references are included herein by reference. Both membrane-bound FLT3 and soluble ligands bind, dimerize and then activate FLT3 kinase.
“Normal cells that express FLT3 Kinase include immature cells of the hematopoietic cell line, which are typically CD34+ cells and brain (see Rosnet, Rosnet, and Small, Blood (1993), 8:1110-19; Small, et.al., Proc. Natl. Acad. Sci. Sci. But, FLT3 kinase is not sufficient to stimulate proliferation. The critical role of FLT3 kinase in immune function is also played by its regulation of dendritic cells proliferation and differentiation (see McKenna, Blood (2000) 95:3489-97; the entire teachings are included herein by reference).
“Numerous hematologic malignancies are FLT3-expressing, with the most prominent being AML (see Yokota and al., Leukemia (1997) 11:1605-09; the entire teachings are included herein by reference). Other FLT3-expressing malignancies include myelodysplastic and T-cell acutely lymphoblastic lesions, B-precursor cells acute lymphoblastic lymphomas, and chronic myelogenous lesions (see Rasko, Leukemia (1995), 9, 2058-66; the entire teachings are included herein).
“Activating mutations are caused by FLT3 kinase mutations that are associated with hematologic malignancies. The FLT3 kinase activates constitutively without binding or dimerization of FLT3 Ligand. This stimulates cells to grow continually. There are two types of activating mutations: internal tandem duplications, (ITDs), and point mutations in the activating loop. The term “FLT3 kinase” is used herein. This term refers to both the wild type FLT3 Kinase as well as mutant FLT3 Kinases such activating mutations-prone FLT3kinases.
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