CATALYTIC INHIBITOR OF PROTEIN PHOSPHATASE 5 ACTIVATES THE EXTRINSIC APOPTOTIC PATHWAY BY DISRUPTING COMPLEX II

Abstract

Protein phosphatase 5 (PP5) is a serine/threonine protein phosphatase involved in the maturation and activation of numerous signaling pathways essential for cancer growth. PP5 activity is essential for the survival of clear cell renal cell carcinoma (ccRCC), however the mechanism remains unclear. Data demonstrates that PP5 interacts with caspase-8, FADD, and RIPK1, components of extrinsic apoptotic pathway Complex II. Specifically, PP5 dephosphorylates and inactivates the death effector proteins RIPK1 and FADD, preserving Complex II integrity and regulating extrinsic apoptosis. Protein phosphatases are considered to be undruggable, however we have developed a specific inhibitor of PP5 (P-053) that prevents substrate binding to the active site. Encouragingly, PP5 inhibition using P-53 in VHL-null ccRCC robustly induces extrinsic apoptosis. Taken together, the data suggests that PP5 promotes ccRCC survival by suppressing extrinsic apoptosis, and small molecule inhibition of PP5 presents a viable therapeutic strategy for ccRCC.

Claims

1. A method of treating cancer, comprising: a) providing i) a subject with cancer, ii) one or more protein phosphatase-5 inhibitors, and b) treating said subject with the one or more PP5 inhibitors.

2. The method of claim 1, wherein treating comprises administering a therapeutically effective amount of one or more protein phosphatase-5 inhibitors to the subject.

3. The method of claim 1, wherein the cancer is renal cell carcinoma.

4. The method of claim 3, wherein the renal cancer is clear cell renal cell carcinoma.

5. The method of claim 1, wherein the one or more protein phosphatase-5 inhibitors are characterized as a PP5 specific inhibitor.

6. The method of claim 1, wherein the one or more protein phosphatase-5 inhibitors are selected from the group consisting of: ##STR00072## ##STR00073## ##STR00074## ##STR00075## or a pharmaceutically acceptable salt thereof, and combinations thereof.

7. The method of claim 1, wherein treating the subject with the one or more protein phosphatase-5 inhibitors is sequential.

8. The method of claim 1, wherein treating the subject with the one or more protein phosphatase-5 inhibitors is simultaneous.

9. The method of claim 1, wherein treating with the one or more protein phosphatase-5 inhibitors results in reduced proliferation of at least some of the cancer cells within said subject.

10. A method of treating cancer, comprising: a) providing i) a subject with cancer and one or more compounds of: ##STR00076## or a pharmaceutically acceptable salt thereof, ##STR00077## or a pharmaceutically acceptable salt thereof, and b) treating the subject with the one or more compounds or pharmaceutically acceptable salts thereof.

11. A method of treating cancer, comprising: a) providing i) a subject with cancer and one or more compounds of: ##STR00078## ##STR00079## ##STR00080## ##STR00081## ##STR00082## and b) treating the subject with the one or more compounds or pharmaceutically acceptable salts thereof.

11. A pharmaceutical anticancer composition comprising one or more PP5 inhibitors, or pharmaceutically acceptable salts thereof.

12. The pharmaceutical anticancer composition of claim 11, wherein the composition is characterized as PP5 specific.

13. The pharmaceutical anticancer composition of claim 11, wherein the one or more PP5 inhibitors are selected from the group consisting of: ##STR00083## ##STR00084## ##STR00085## ##STR00086## ##STR00087## or a pharmaceutically acceptable salt thereof, and combinations thereof.

14. A pharmaceutical anticancer composition comprising one or more PP5 inhibitors selected from the group consisting of: ##STR00088## ##STR00089## ##STR00090## ##STR00091## ##STR00092## and combinations thereof.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0013] The patent or application file contains at least one drawing executed in color. Copies of this patent or patent application publication with color drawing(s) will be provided by the Office upon request and payment of the necessary fee.

[0014] FIGS. 1A-1H present the identification and characterization of small molecule inhibitors of PP5. FIG. 1A presents a schematic workflow of in silico screening a library of 3.7-million compounds to identify potential PP5 inhibitors. A single overlapping hit, P0 was used to identify a set of analogs for cell-based screening. Compounds 5 (P5) and 13 (P13) were selected as candidate PP5 inhibitors. FIG. 1B presents 786-O cells were treated with the indicated amounts of P5 and P13 for 48 hrs. The effect of PP5 inhibitors on cell viability was assessed by MTT assay. Error bars represent the standard deviation (S.D.) of three independent experiments. A Student's t-test was performed to assess statistical significance **P<0.01; ***P<0.001. FIG. 1C presents 786-O cells were treated with 10 M P5 or P13 or DMSO vehicle control for 24 hrs. Induction of apoptosis was assessed by immunoblotting. GAPDH was used as a loading control. FIG. 1D presents a PP5 enzyme kinetic data with P5 (orange line), P13 (red line) or no inhibitor (blue line) presented as a Lineweaver-Burk plot (n=3 independent samples). FIG. 1E presents Lysates from HEK293 cells transfected with PP5-FLAG were incubated with indicated amounts of biotin-labeled P5 and P13 followed by streptavidin agarose pulldown. Co-pulldown of PP5-FLAG was detected by immunoblotting. FIG. 1F presents crystal structure of PP5 active site (gray; PDB:3H60) with predicted residues responsible (green) for contact with P13 (orange) modeled with PyMOL software (v4.6.0). FIG. 1G presents T-PP5-FLAG and predicted binding residue mutants were transiently expressed in HEK293 cells. Lysate was incubated with indicated amounts of biotinylated P13 followed by streptavidin pulldown (above). PP5 binding to biotinylated P13 was examined by immunoblot. Input expression of PP5-FLAG and mutants is below. EV was used as a control. FIG. 1H presents P13-BODIPY binding to PP5 measured by fluorescence anisotropy. Points are replicate meansSEM from independent trials for display. (n=2).

[0015] FIGS. 2A-2H present inhibitory effect of P053 compound on PP5 and induction apoptosis in VHL-null ccRCC. FIG. 2A presents 786-O cells were treated with 10 M of the indicated second generation of PP5 inhibitors for 24 hrs. DMSO was used as control. PP5 inhibition was evaluated by immunoblotting for the bono fide substrate phospho-S211-GR. SE (short exposure) and LE (long exposure) of the radiographic film. GAPDH was used as a loading control. FIG. 2B presents 786-O cells were again treated with 10 M of the indicated second generation of PP5 inhibitors for 24 hrs. DMSO was used as control. Induction of apoptosis was evaluated by immunoblotting using apoptotic markers cleaved PARP and cleaved caspase 8. GAPDH was used as a loading control. GIG 2C presents 786-O cells were treated with indicated amounts of P13, P053, P058, P059, and P075 for 24 hrs and then cell proliferation was assessed by the MTT assay. A Student's t-test was performed to assess statistical significance *P<0.05; **P<0.01 or non-significant (ns). FIG. 2D presents PP5 enzyme kinetic data with P053 (purple) or no inhibitor (blue line) presented as a Lineweaver-Burk plot (n=3 independent samples). FIG. 2E presents the structure of PP5 catalytic domain (gray; PDB 3H60) bound to PP5 inhibitors P5 (green), P13 (orange), and P053 (yellow). FIG. 2F presents lysate from 786-O cells was collected and incubated with 1 M biotinylated P5 or P13 for 1 hr and then challenged with 1 M P053 for 30 min. Streptavidin-coated agarose beads were used to pull down the biotinylated compounds. Co-pulldown of endogenous PP5 and PP2A was assessed by immunoblotting. SE (short exposure) and LE (long exposure) of the radiographic film. FIG. 2G presents VHL containing HK2, Caki-2, and Caki-1 cells and VHL-null cells, A498 and 786-O, were treated with the indicated amount of P053 for 24 hrs, and then cell proliferation was assessed by the MTT assay. A Student's t-test was performed to assess statistical significance *P<0.05; **P<0.01 or non-significant (ns). FIG. 2H presents Caki-1, Caki-2, A498, and 786-O cells were treated with 30 or 40 M P053 or DMSO control (0 M) for 24 hrs. Following propidium iodide (PI) staining cell death was assessed by flow cytometry. Percentage of PI stained cells was normalized to the vehicle control for each cell line individually.

[0016] FIGS. 3A-3H present targeting PP5 activates the extrinsic apoptotic pathway. FIG. 3A presents ccRCC 786-O cells were treated with 10 M compound P053 for 24 hrs. Induction of apoptosis was evaluated by immunoblotting using apoptotic markers as indicated. GAPDH was used as a loading control. FIG. 3B presents PP5 was silenced by small interfering RNA (siRNA) in VHL-null ccRCC cells 786-O and A498. Induction of apoptosis was evaluated by immunoblotting using apoptotic markers as indicated. siCtrl represents the non-targeting siRNA control. GAPDH was used as a loading control. FIG. 3C presents Inhibition of CK1 by indicated amounts of IC261 for 16 hrs in 786-O cells. Induction of apoptotic markers was assessed by immunoblotting. GAPDH was used as a loading control. FIG. 3D presents ccRCC 786-O cells were treated in presence (+) or absence () of 10 M apoptotic inhibitor z-VAD-fmk for 1 hr followed by the addition of indicated amounts of CK1 inhibitor, IC261, for an additional 16 hrs. Induction of apoptosis was evaluated by the immunoblotting. GAPDH was used as a loading control. FIG. 3E presents a schematic representation of the extrinsic apoptotic pathway. Death receptors are activated by binding of death ligands. The leads to binding of adaptors and ultimately formation of complex II containing FADD, RIPK1, and pro-caspase 8. Upon complex II formation, caspase 8 is activated and then released from the complex leading to downstream caspase induction and apoptosis. FIG. 3F presents an endogenous PP5 was immunoprecipitated (IP) from 786-O cells. Coimmunoprecipitation (co-IP) of FADD, RIPK1, and caspase 8 was examined by immunoblot. IgG was used as a control. GAPDH was used as a loading control. FIG. 3G presents WT-PP5-FLAG was transiently expressed and isolated from 786-O cells. Co-IP of FADD and GR was examined by immunoblotting. Phosphorylation level of S194-FADD was assessed by immunoblot. PP5 activity was evaluated by immunoblotting for the bono fide substrate phospho-S211-GR as a control. GAPDH was used as a loading control. FIG. 3H presents CK1 was inhibited with indicated amounts of IC261 for 24 hrs in ccRCC cells 786-O and A498. Induction of apoptotic markers shown by immunoblotting using anti-cleaved caspase-3 antibody. Phosphorylation of S194-FADD was evaluated by western blot. GAPDH was used as a loading control.

[0017] FIGS. 4A-4H present PP5 mediates complex II formation. FIG. 4A presents endogenous FADD (left) and RIPK1 (right) were IP from WT-HAP1 and PP5-KO HAP1 cells. Co-IPs of RIPK1 and FADD were examined by immunoblot. GAPDH was used as a loading control. FIG. 4B presents endogenous FADD (left) and PP5 (right) were IP from WT-HAP1 and RIPK1-KO HAP1 cells. Co-IPs of PP5 and FADD were examined by immunoblot. GAPDH was used as a loading control. FIG. 4C presents endogenous RIPK1 (left) and PP5 (right) were IP from WT-HAP1 and FADD-KO HAP1 cells. Co-IPs of PP5 and RIPK1 were examined by immunoblot. GAPDH was used as a loading control. FIG. 4D presents FADD-FLAG and death domain deleted FADD (FADD-FLAG-ADD) were transiently transfected and IP from HEK293 cells. EV was used as a control. Co-IP of RIPK1 and PP5 was examined by immunoblot. GAPDH was used as a loading control. FIG. 4E presents RIPK1-HA and death domain deleted RIPK1 (RIPK1-HA-ADD) were transiently transfected and isolated from HEK293 cells. EV was used as a control. Co-IP of FADD and PP5 was examined by immunoblot. GAPDH was used as a loading control. FIG. 4F presents HEK293 cells were treated with 2 M SNX-2112 for the indicated times. FADD and RIPK1 protein levels were examined by immunoblot. Bona fide Hsp90 clients Tsc2, Akt and phos-Ser473-Akt were used as a positive control. GAPDH was used as a loading control. FIG. 4G presents FADD was IP from 786-O cells. Co-IP of PP5 and Hsp90 was evaluated by immunoblot. IgG was used as a negative control. GAPDH was used as a loading control. FIG. 4H presents a schematic illustration of PP5-mediated downregulation of the extrinsic apoptotic pathway. On the left, in the absence of PP5, following activation of the death receptors and complex II formation, caspase 8 is cleaved resulting in activation of apoptosis. On the right, when PP5 associates with complex II and dephosphorylates FADD on S194, the downstream apoptotic pathway is suppressed and leads to ccRCC cell survival.

[0018] FIGS. S1A-S1F present development of specific inhibitors of PP5 (Related to FIG. 1). FIG. S1A presents In silico docking study used to screen a library of 3.7 million compounds using PP5 crystal structure (PDB:3H60). Top-ranked hits were filtered and used for in vitro experiments. Lead compounds are shown bound to PP5 at the final step. FIG. S1B presents P0 inhibitor of PP5 (PDB:3H60) visualized on PyMOL v4.6.0. FIG. S1C presents 786-O cells were treated with indicated amounts of P0 for 24 hrs. DMSO was used as a control. PP5 inhibition was evaluated by immunoblotting for the bono fide substrates phospho-S211-GR and phospho-S13-Cdc37. GAPDH was used as a loading control. FIG. S1D presents chemical structures of 14 identified candidate PP5 inhibitors based on P0 that were used in cell-based screening. FIG. S1E presents 786-O cells were treated with 20 M P0-P13 for 24 hrs. DMSO was used as a control. Induction of apoptosis was assessed by immunoblotting using cleaved caspase 3. PP5 inhibition was evaluated by immunoblotting for the bono fide substrates phospho-S211-GR and phospho-S13-Cdc37. GAPDH was used as a loading control. FIG. S1F presents 786-O cells were treated with the indicated amounts of selected PP5 inhibitors for 48 hrs. The effect of PP5 inhibitors on cell viability was assessed by MTT assay. Errors bars represent the meanS.D. of three independent experiments. A Student's t-test was performed to assess statistical significance. *P<0.05; **P<0.01; ***P<0.001; ****P<0.0001.

[0019] FIGS. S2A-S2F present chemical synthesis of PP5 inhibitors (Related to FIG. 1). FIG. S2A presents .sup.1H NMR of P5. FIG. S2B presents .sup.13C NMR of P5. FIG. S2C presents .sup.1H NMR of P13. FIG. S2D presents .sup.13C NMR of P13. FIG. S2E presents synthesis of biotin conjugated P5. FIG. S2F presents synthesis of biotin conjugated P13.

[0020] FIGS. S3A-S3F present chemical synthesis of fluorescently labeled P13 second generation of PP5 inhibitors (Related to FIGS. 1-2). FIG. S3A presents synthesis and structure of BODIPY-labeled P13. FIG. S3B presents coomassie-stained SDS PAGE gel showing purified PP5 protein used in FIG. 1H. FIG. S3C presents chemical structures of the second generation PP5 inhibitors derived from P13. FIG. S3D presents synthesis scheme of compounds 16, 17, 18, and 19 from Figure S4A. FIG. S3E presents .sup.1H NMR of P053. FIG. S3F presents .sup.13C NMR of P053.

[0021] FIGS. S4A-S4F present specific small molecule inhibitor of PP5 induces apoptosis in VHL-null ccRCC (Related to FIGS. 2-3). FIG. S4A presents IC.sub.50 measurements for PP5 in the presence of P5 (orange line), P13 (red line), or P053 (purple line) (n=2). Data shown as meanS.D. FIG. S4B presents VHL protein from HEK293, HK2, Caki-1, Caki-2, A498 and 786-O cells was assessed by immunoblotting. GAPDH was used as a loading control. FIG. S4C presents Flow-cytometric assessment of cell death in Caki-1, Caki-2, A498, and 786-O following treatment with 30 M P053 for 24 hrs (blue) or 40 M for 24 hrs (orange), as determined by PI staining. Pink indicates no treatment (NT). FIG. S4D presents targeted siRNA was used to knockdown (KD) PP5 in 786-O cells. PP5 KD was confirmed by immunoblotting. Induction of apoptotic markers shown by immunoblotting using anti-cleaved caspase-3 and -8 and cleaved PARP antibodies. Phosphorylation level of S194-FADD was assessed by immunoblot. GAPDH was used as a control. FIG. S4E presents increased phosphorylation of S194-FADD in PP5 KO HAP1 cells was assessed by immunoblotting. GAPDH was used as a control. FIG. S4F presents WT-PP5-FLAG was transiently expressed in 786-O cells. EV was used as a control. Phosphorylation level of S161-RIPK1 and S166-RIPK1 was assessed by immunoblot. GAPDH was used as a loading control.

[0022] It is noted that the drawings of the disclosure are not necessarily to scale. The drawings are intended to depict only typical aspects of the disclosure, and therefore should not be considered as limiting the scope of the disclosure. In the drawings, like numbering represents like elements between the drawings.

Definitions

[0023] As used in the present specification, the following words and phrases are generally intended to have the meanings as set forth below, except to the extent that the context in which they are used indicates otherwise.

[0024] As used herein, the singular forms a, an, and the include plural references unless the context clearly dictates otherwise. Thus, for example, references to a compound include the use of one or more compound(s). A step of a method means at least one step, and it could be one, two, three, four, five or even more method steps.

[0025] As used herein the terms about, approximately, and the like, when used in connection with a numerical variable, generally refers to the value of the variable and to all values of the variable that are within the experimental error (e.g., within the 95% confidence interval [CI 95%] for the mean) or within 10% of the indicated value, whichever is greater.

[0026] As used herein the term alkyl refers to C.sub.1-20 inclusive, linear (i.e., straight-chain), branched, or cyclic, saturated or at least partially and in some cases fully unsaturated (i.e., alkenyl and alkynyl) hydrocarbon chains, including for example, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, pentyl, hexyl, octyl, ethenyl, propenyl, butenyl, pentenyl, hexenyl, octenyl, butadienyl, propynyl, butynyl, pentynyl, hexynyl, heptynyl, and allenyl groups. Branched refers to an alkyl group in which a lower alkyl group, such as methyl, ethyl or propyl, is attached to a linear alkyl chain. Lower alkyl refers to an alkyl group having 1 to about 8 carbon atoms (i.e., a C.sub.1-8 alkyl), e.g., 1, 2, 3, 4, 5, 6, 7, or 8 carbon atoms. Higher alkyl refers to an alkyl group having about 10 to about 20 carbon atoms, e.g., 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 carbon atoms. In certain embodiments, alkyl refers to C.sub.1-8 straight-chain alkyls. In other embodiments, alkyl refers to C.sub.1-8 branched-chain alkyls. In embodiments, alkyl groups can optionally be substituted (a substituted alkyl) with one or more alkyl group substituents, which can be the same or different. The term alkyl group substituent includes but is not limited to alkyl, substituted alkyl, halo, arylamino, acyl, hydroxyl, aryloxyl, alkoxyl, alkylthio, arylthio, aralkyloxyl, aralkylthio, carboxyl, alkoxycarbonyl, oxo, and cycloalkyl. There can be optionally inserted along the alkyl chain one or more oxygen, sulfur or substituted or unsubstituted nitrogen atoms, wherein the nitrogen substituent is hydrogen, lower alkyl (also referred to herein as alkylaminoalkyl), or aryl. Thus, as used herein, the term substituted alkyl includes alkyl groups, as defined herein, in which one or more atoms or functional groups of the alkyl group are replaced with another atom or functional group, including for example, alkyl, substituted alkyl, halogen, aryl, substituted aryl, alkoxyl, hydroxyl, nitro, amino, alkylamino, dialkylamino, sulfate, and mercapto.

[0027] As used herein, the term heteroalkyl by itself or in combination with another term means, unless otherwise stated, a stable straight or branched chain alkyl group consisting of the stated number of carbon atoms and one or two heteroatoms of O, N, and S, and wherein the nitrogen and sulfur atoms may be optionally oxidized and the nitrogen heteroatom may be optionally quaternized. In embodiments, the heteroatom(s) may be placed at any position of the heteroalkyl group, including between the rest of the heteroalkyl group and the fragment to which it is attached, as well as attached to the most distal carbon atom in the heteroalkyl group. Non-limiting examples include: OCH.sub.2CH.sub.2CH.sub.3, CH.sub.2CH.sub.2CH.sub.2OH, CH.sub.2CH.sub.2CH.sub.2NHCH.sub.3, and CH.sub.2SCH.sub.2CH.sub.3. In embodiments, up to two heteroatoms may be consecutive, such as, for example, CH.sub.2NHOCH.sub.3, or CH.sub.2CH.sub.2SSCH.sub.3. In embodiments, heteroalkyl groups have 1-12 carbons.

[0028] As used herein, the term alkenyl, denotes a monovalent group derived from a hydrocarbon moiety containing at least two carbon atoms and at least one carbon-carbon double bond. In embodiments, the double bond may or may not be the point of attachment to another group. Alkenyl groups (e.g., C.sub.2-C.sub.8-alkenyl) include, but are not limited to, for example, ethenyl, propenyl, prop-1-en-2-yl, butenyl, 1-methyl-2-buten-1-yl, heptenyl, octenyl and the like.

[0029] As used herein the term apoptosis, refers to a form of programmed cell death in multicellular organisms that involves a series of biochemical events that lead to a variety of morphological changes, including blebbing, changes to the cell membrane such as loss of membrane asymmetry and attachment, cell shrinkage, nuclear fragmentation, chromatin condensation, and chromosomal DNA fragmentation. Defective apoptotic processes have been implicated in an extensive variety of diseases; for example, defects in the apoptotic pathway have been implicated in diseases associated with uncontrolled cell proliferations, such as cancer. See e.g. U.S. Patent Publication No. 20200289513 herein entirely incorporated by reference.

[0030] A number of terms herein relate to cancer. Cancer is intended herein to encompass all forms of abnormal or improperly regulated reproduction of cells in a subject.

[0031] The growth of cancer cells (growth herein referring generally to cell division but also to the growth in size of masses of cells) is characteristically uncontrolled or inadequately controlled, as is the death (e.g., apoptosis) of such cells. Local accumulations of such cells result in a tumor. More broadly, and still denoting tumors herein are accumulations ranging from a cluster of lymphocytes at a site of infection to vascularized overgrowths, both benign and malignant. A malignant tumor (as opposed to a benign tumor) herein includes cells that tend to migrate to nearby tissues, including cells that may travel through the circulatory system to invade or colonize tissues or organs at considerable remove from their site of origin in the primary tumor, so-called herein. Metastatic cells are adapted to penetrate blood vessel wells to enter (intravasate) and exit (extravasate) blood vessels. Tumors capable of releasing such cells are also referred to herein as metastatic. The term is used herein also to denote any cell in such a tumor that is capable of such travel, or that is en route, or that has established a foothold in a target tissue. For example, a metastatic breast cancer cell that has taken root in the lung is referred to herein as a lung metastasis. Metastatic cells may be identified herein by their respective sites of origin and destination, such as breast-to-bone metastatic. In the target tissue, a colony of metastatic cells can grow into a secondary tumor, so called herein.

[0032] The terms modified, mutant, and variant (when the context so admits) refer to a gene or gene product that displays modifications in sequence and or functional properties (i.e., altered characteristics) when compared to the wild-type gene or gene product. In some embodiments, the modification includes at least one nucleotide insertion, deletion, or substitution.

[0033] The terms carrier and vehicle as used herein refer to usually inactive accessory substances into which a pharmaceutical substance is suspended. Exemplary carriers include liquid carriers (such as water, saline, culture medium, saline, aqueous dextrose, and glycols) and solid carriers (such as carbohydrates exemplified by starch, glucose, lactose, sucrose, and dextrans, anti-oxidants exemplified by ascorbic acid and glutathione, and hydrolyzed proteins.

[0034] As used herein, the term halo or halogen alone or as part of another substituent means, unless otherwise stated, a fluorine, chlorine, bromine, or iodine atom.

[0035] Cyclic and cycloalkyl refer to a non-aromatic mono- or multicyclic ring system of about 3 to about 10 carbon atoms, e.g., 3, 4, 5, 6, 7, 8, 9, or 10 carbon atoms. In embodiments, a cycloalkyl group can be optionally partially unsaturated. In embodiments, the cycloalkyl group also can be optionally substituted with an alkyl group substituent as defined herein, oxo, and/or alkylene. In embodiments, there can be optionally inserted along the cyclic alkyl chain one or more oxygen, sulfur or substituted or unsubstituted nitrogen atoms, wherein the nitrogen substituent is hydrogen, alkyl, substituted alkyl, aryl, or substituted aryl, thus providing a heterocyclic group. Non-limiting examples of monocyclic cycloalkyl rings include cyclopentyl, cyclohexyl, and cycloheptyl. Non-limiting examples of multicyclic cycloalkyl rings include adamantyl, octahydronaphthyl, decalin, camphor, camphane, and noradamantyl.

[0036] As used herein, the term heterocycloalkyl or heterocyclyl refers to a heteroalicyclic group including one to four ring heteroatoms each selected from O, S, and N. In embodiments, each heterocyclyl group has from 3 to 10 atoms in its ring system, with the proviso that the ring of said group does not contain two adjacent O or S atoms. In embodiments, heterocyclyl substituents may be alternatively defined by the number of carbon atoms, e.g., C.sub.2-C.sub.8-heterocyclyl indicates the number of carbon atoms contained in the heterocyclic group without including the number of heteroatoms. For example, a C.sub.2-C.sub.8-heterocyclyl will include an additional one to four heteroatoms. In embodiments, the heterocyclyl group has less than three heteroatoms. In embodiments, the heterocyclyl group has one to two heteroatoms. In embodiments, the heterocycloalkyl group is fused with an aromatic ring. In embodiments, nitrogen and sulfur heteroatoms may be optionally oxidized, and the nitrogen atom may be optionally quaternized. The heterocyclic system may be attached, unless otherwise stated, at any heteroatom or carbon atom that affords a stable structure.

[0037] As used herein, the term aromatic refers to a carbocycle or heterocycle with one or more polyunsaturated rings and having aromatic character, i.e., having (4n+2) delocalized (pi) electrons, where n is an integer.

[0038] The term aryl is used herein to refer to an aromatic substituent that can be a single aromatic ring, or multiple aromatic rings that are fused together, linked covalently, or linked to a common group, such as, but not limited to, a methylene or ethylene moiety. The common linking group also can be a carbonyl, as in benzophenone, or oxygen, as in diphenylether, or nitrogen, as in diphenylamine. In embodiments, the term aryl specifically encompasses heterocyclic aromatic compounds. The aromatic ring(s) can include phenyl, naphthyl, biphenyl, diphenylether, diphenylamine and benzophenone, among others. In particular embodiments, the term aryl means a cyclic aromatic including about 5 to about 10 carbon atoms, e.g., 5, 6, 7, 8, 9, or 10 carbon atoms, and including 5- and 6-membered hydrocarbon and heterocyclic aromatic rings. In embodiments, an aryl group can be optionally substituted (a substituted aryl) with one or more aryl group substituents, which can be the same or different, wherein aryl group substituent includes alkyl, substituted alkyl, aryl, substituted aryl, aralkyl, hydroxyl, alkoxyl, aryloxyl, aralkyloxyl, carboxyl, acyl, halo, nitro, alkoxycarbonyl, aryloxycarbonyl, aralkoxycarbonyl, acyloxyl, acylamino, aroylamino, carbamoyl, alkylcarbamoyl, dialkylcarbamoyl, arylthio, alkylthio, alkylene, and NRR, wherein R and R can each be independently hydrogen, alkyl, substituted alkyl, aryl, substituted aryl, and aralkyl. Thus, as used herein, the term substituted aryl includes aryl groups, as defined herein, in which one or more atoms or functional groups of the aryl group are replaced with another atom or functional group, including for example, alkyl, substituted alkyl, halogen, aryl, substituted aryl, alkoxyl, hydroxyl, nitro, amino, alkylamino, dialkylamino, sulfate, and mercapto. Non-limiting examples of aryl groups include, but are not limited to, cyclopentadienyl, phenyl, furan, thiophene, pyrrole, pyran, pyridine, imidazole, benzimidazole, isothiazole, isoxazole, pyrazole, pyrazine, triazine, pyrimidine, quinoline, isoquinoline, indole, carbazole, and the like.

[0039] As generally discussed herein, a structure represented generally by the formula:

##STR00007##

as used herein refers to a ring structure, for example, but not limited to a 3-carbon, a 4-carbon, a 5-carbon, a 6-carbon, and the like, aliphatic and/or aromatic cyclic compound, including a saturated ring structure, a partially saturated ring structure, and an unsaturated ring structure as defined herein, including a substituent R group. In embodiments, the R group can be present or absent, and when present, one or more R groups can each be substituted on one or more available carbon atoms of the ring structure. The presence or absence of the R group and number of R groups is determined by the value of the integer n. Each R group, if more than one, is substituted on an available carbon of the ring structure rather than on another R group. For example, the structure above where n is 0 to 2 would comprise compound groups including, but not limited to:

##STR00008##

and the like.

[0040] In embodiments, a dashed line representing a bond in a cyclic ring structure indicates that the bond can be either present or absent in the ring. That is a dashed line representing a bond in a cyclic ring structure indicates that the ring structure is one of a saturated ring structure, a partially saturated ring structure, and an unsaturated ring structure.

[0041] In some embodiments, the compounds described by the presently disclosed subject matter contain a linking group. As used herein, the term linking group includes a chemical moiety, such as a furanyl, phenylene, thienyl, and pyrrolyl radical, which is bonded to two or more other chemical moieties, in particular aryl groups, to form a stable structure.

[0042] In embodiments, a named R, or L, group will generally have the structure that is recognized in the art as corresponding to a group having that name, unless specified otherwise herein. These definitions are intended to supplement and illustrate, not preclude, the definitions that would be apparent to one of ordinary skill in the art upon review of the present disclosure.

[0043] As used herein, the term target activity refers to a biological activity capable of being modulated by a selective modulator. Certain exemplary target activities include, but are not limited to, binding affinity, signal transduction, enzymatic activity, tumor growth, inflammation or inflammation-related processes, or amelioration of one or more symptoms associated with a disease or condition such as a disease or condition relating to cancer.

[0044] As used herein, the term target protein refers to a molecule or a portion of a protein capable of being bound by a selective binding compound. For example PP5 is a target protein for the PP5 inhibitor compositions or selective binding compounds of the present disclosure.

[0045] As used herein, the term treatment or treating is defined as the application or administration of a therapeutic agent, i.e., a one or more polycyclic compounds of the present disclosure such as one or more PP5 inhibitors, or pharmaceutically acceptable salts or solvates thereof, (alone or in combination with another pharmaceutical agent, carrier, or vehicle), to a patient, or application or administration of a therapeutic agent.

[0046] A therapeutically effective amount is that amount that will generate the desired therapeutic outcome (i.e., achieve therapeutic efficacy). For example, a therapeutically effective dose of a compound of the present disclosure is an amount that is sufficient to palliate, ameliorate, stabilize, reverse, prevent, slow or delay the progression of the disease state (e.g., cancer). A therapeutically effective amount can be an amount administered in a dosage protocol that includes days or weeks of administration. In certain embodiments, a therapeutically effective dose of a compound is able to improve at least one sign or symptom of a disease state. As used herein, the terms effective amount, and pharmaceutically effective amount, have the same meaning as therapeutically effective amount. In embodiments, a therapeutically effective amount alters the natural state of a subject.

[0047] As used herein, the term patient, individual or subject refers to a human or a non-human mammal. Non-human mammals include, for example, livestock and pets, such as ovine, bovine, porcine, canine, feline and murine mammals. Non-human mammals also include non-human primates, rats, rabbits and camelids. In certain embodiments, the patient, subject, or individual is human.

[0048] As used herein, the phrases selective inhibition or selectively inhibit refer to a molecule's ability to inhibit the activity or expression of a particular protein or protein isoform, or RNA, while being unable to inhibit the protein activity or expression of another protein or protein isoform, or RNA, by more than 5%. In embodiments, the PP5 inhibitors of the present disclosure selectively inhibit PP5.

[0049] As used herein, the term pharmaceutically acceptable refers to a material, such as a carrier or diluent, which does not abrogate the biological activity or properties of the compound, and is relatively non-toxic, i.e., the material may be administered to an individual without causing undesirable biological effects or interacting in a deleterious manner with any of the components of the composition in which it is contained.

[0050] As used herein, the term pharmaceutically acceptable salt refers to derivatives of one or more of the polycyclic compounds of the present disclosure such as PP5 inhibitors, wherein the parent compound is modified by converting an existing acid or base moiety to its salt form. Examples of pharmaceutically acceptable salts include, but are not limited to, mineral or organic acid salts of basic residues such as amines; alkali or organic salts of acidic residues such as carboxylic acids; and the like. The pharmaceutically acceptable salts of the present disclosure include the conventional non-toxic salts of the parent compound formed, for example, from non-toxic inorganic or organic acids. The pharmaceutically acceptable salts of the present invention can be synthesized from the parent compound which contains a basic or acidic moiety by conventional chemical methods. Generally, such salts can be prepared by reacting the free acid or base forms of these compounds with a stoichiometric amount of the appropriate base or acid in water or in an organic solvent, or in a mixture of the two; generally, nonaqueous media like ether, ethyl acetate, ethanol, isopropanol, or acetonitrile are preferred. Lists of suitable salts are found in Remington's Pharmaceutical Sciences, 17th ed., Mack Publishing Company, Easton, Pa., 1985, p. 1418 and Journal of Pharmaceutical Science, 66, 2 (1977), each of which is incorporated herein by reference in its entirety.

[0051] As used herein, the term solvate refers to complexes of the compounds disclosed herein or salts thereof with solvent molecules, e.g. organic solvent molecules and/or water.

[0052] General methods in molecular and cellular biochemistry can be found in such standard textbooks as Molecular Cloning: A Laboratory Manual, 3rd Ed. (Sambrook et al., Harbor Laboratory Press 2001); Short Protocols in Molecular Biology, 4th Ed. (Ausubel et al. eds., John Wiley & Sons 1999); Protein Methods (Bollag et al., John Wiley & Sons 1996); Nonviral Vectors for Gene Therapy (Wagner et al. eds., Academic Press 1999); Viral Vectors (Kaplift & Loewy eds., Academic Press 1995); Immunology Methods Manual (I. Lefkovits ed., Academic Press 1997); and Cell and Tissue Culture: Laboratory Procedures in Biotechnology (Doyle & Griffiths, John Wiley & Sons 1998), the disclosures of which are incorporated herein by reference. In embodiments, there may be employed conventional molecular biology, microbiology, and recombinant DNA techniques within the skill of the art. Such techniques are explained fully in the literature. See, e.g., Sambrook et al, 2001, Molecular Cloning: A Laboratory Manual; Ausubel, ed., 1994, Current Protocols in Molecular Biology Volumes I-III; Celis, ed., 1994, Cell Biology: A Laboratory Handbook Volumes I-III; Coligan, ed., 1994, Current Protocols in Immunology Volumes I-III; Gait ed., 1984, Oligonucleotide Synthesis; Hames & Higgins eds., 1985, Nucleic Acid Hybridization; Hames & Higgins, eds., 1984, Transcription And Translation; Freshney, ed., 1986, Animal Cell Culture; IRL Press, 1986, Immobilized Cells And Enzymes; Perbal, 1984, A Practical Guide To Molecular Cloning.

[0053] Before embodiments are further described, it is to be understood that this disclosure is not limited to particular embodiments described, as such may, of course, vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting.

[0054] Where a range of values is provided, it is understood that each intervening value, to the tenth of the unit of the lower limit unless the context clearly dictates otherwise, between the upper and lower limit of that range and any other stated or intervening value in that stated range, is encompassed within the invention. The upper and lower limits of these smaller ranges may independently be included in the smaller ranges, and are also encompassed within the invention, subject to any specifically excluded limit in the stated range. Where the stated range includes one or both of the limits, ranges excluding either or both of those included limits are also included in the invention.

[0055] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although any methods and materials similar or equivalent to those described herein can also be used in the practice or testing of the present invention, the preferred methods and materials are now described. All publications mentioned herein are incorporated herein by reference to disclose and describe the methods and/or materials in connection with which the publications are cited.

DETAILED DESCRIPTION OF THE EMBODIMENTS

[0056] Throughout the specification and claims, a given chemical formula or name shall encompass all optical and stereoisomers, as well as racemic mixtures where such isomers and mixtures exist.

[0057] The present disclosure provides therapeutic and preventative agents for treating or preventing diseases, disorders and conditions involving cancer. Non-limiting examples of diseases, disorders and conditions involving cancer such as kidney cancer, include one or more renal cell carcinomas (RCC) such as clear cell (ccRCC), papillary, chromophobe, collecting duct, or otherwise unclassified renal cell carcinoma. In embodiments, conditions for treatment include metastasis.

[0058] In embodiments, the present disclosure includes one or more polycyclic compounds, or pharmaceutically acceptable salts or solvates thereof, including: one or more PP5 inhibitors, such as, for example:

##STR00009## ##STR00010##

[0059] In embodiments, the one or more compounds target complex Ila and/or induce the extrinsic apoptotic pathway in cancer cells. For example, the compounds of the present disclosure destabilize complex Ila, which typically includes PP5, FADD, ProCasp8, and RIPK1 constituents.

[0060] Advantages of the present disclosure include compositions and methods for treating, ameliorating, or preventing cancer. In embodiments, compositions are provided that are pharmaceutically acceptable. In embodiments, compositions of the present disclosure target, bind, and/or selectively bind to PP5. In embodiments, compositions of the present disclosure induce the extrinsic apoptotic pathway in cancer cells.

[0061] In embodiments, a compound of the present disclosure is

##STR00011##

or a pharmaceutically acceptable salt or solvate thereof.

[0062] In embodiments, a compound of the present disclosure is

##STR00012##

or a pharmaceutically acceptable salt or solvate thereof.

[0063] In embodiments, a compound of the present disclosure is

##STR00013##

[0064] In embodiments, a compound of the present disclosure is

##STR00014##

or a pharmaceutically acceptable salt or solvate thereof.

[0065] In embodiments, a compound of the present disclosure is

##STR00015##

or a pharmaceutically acceptable salt or solvate thereof.

[0066] In embodiments, a compound of the present disclosure is

##STR00016##

or a pharmaceutically acceptable salt or solvate thereof.

[0067] In embodiments, a compound of the present disclosure is

##STR00017##

or a pharmaceutically acceptable salt or solvate thereof.

[0068] In embodiments, a compound of the present disclosure is

##STR00018##

or a pharmaceutically acceptable salt or solvate thereof.

[0069] In embodiments, a compound of the present disclosure is

##STR00019##

or a pharmaceutically acceptable salt or solvate thereof.

[0070] In embodiments, a compound of the present disclosure is

##STR00020##

or a pharmaceutically acceptable salt or solvate thereof, and combinations thereof.

[0071] In embodiments, one or more compounds or formulas as disclosure herein is provided as a pharmaceutically acceptable salt such as derivatives of one or more of the polycyclic compounds of the present disclosure. In embodiments, a parent compound such as a polycyclic compound of the present disclosure is modified by converting an existing acid or base moiety thereof to a salt form. Examples of pharmaceutically acceptable salts include, but are not limited to, mineral or organic acid salts of basic residues such as amines; alkali or organic salts of acidic residues such as carboxylic acids; and the like. In embodiments, the pharmaceutically acceptable salts of the present disclosure include the conventional non-toxic salts of the parent compound formed, for example, from non-toxic inorganic or organic acids. The pharmaceutically acceptable salts of the present disclosure can be synthesized from the parent compound which contains a basic or acidic moiety by conventional chemical methods. Generally, such salts can be prepared by reacting the free acid or base forms of these compounds with a stoichiometric amount of the appropriate base or acid in water or in an organic solvent, or in a mixture of the two; generally, nonaqueous media like ether, ethyl acetate, ethanol, isopropanol, or acetonitrile.

[0072] In embodiments, the polycyclic compounds of the present disclosure may possess one or more stereocenters, and each stereocenter may exist independently in either the R or S configuration.

[0073] It is also to be understood that compounds that have the same molecular formula but differ in the nature or sequence of bonding of their atoms or the arrangement of their atoms in space are termed isomers. Isomers that differ in the arrangement of their atoms in space are termed stereoisomers, for example, diastereomers, enantiomers, and atropisomers. Stereoisomers that are not mirror images of one another are termed diastereomers and those that are non-superimposable mirror images of each other are termed enantiomers. When a compound has an asymmetric center, for example, it is bonded to four different groups, a pair of enantiomers is possible. An enantiomer can be characterized by the absolute configuration of its asymmetric center and is described by the R- and S-sequencing rules of Cahn and Prelog, or by the manner in which the molecule rotates the plane of polarized light and designated as dextrorotatory or levorotatory (i.e., as (+) or ()-isomers respectively). A chiral compound can exist as either individual enantiomer or as a mixture thereof. A mixture containing equal proportions of the enantiomers is called a racemic mixture.

[0074] Unless indicated otherwise, the description or naming of a particular compound in the specification and claims is intended to include both individual enantiomers and mixtures, racemic or otherwise, thereof. Within the present disclosure, any open valency appearing on a carbon, oxygen, or nitrogen atom in any structure described herein indicates the presence of a hydrogen atom. Where a chiral center exists in a structure, but no specific stereochemistry is shown for that center, both enantiomers, separately or as a mixture, are encompassed by that structure. The methods for the determination of stereochemistry and the separation of stereoisomers are well-known in the art.

[0075] In embodiments, the polycyclic compounds may exist as tautomers. All tautomers are included within the scope of the compounds presented herein.

[0076] Compounds described herein also include isotopically-labeled compounds wherein one or more atoms is replaced by an atom having the same atomic number, but an atomic mass or mass number different from the atomic mass or mass number usually found in nature. Non-limiting examples of isotopes suitable for inclusion in the compounds described herein include and are not limited to .sup.2H or deuterium. In one embodiment, isotopically-labeled compounds are useful in drug or substrate tissue distribution studies. In another embodiment, substitution with heavier isotopes such as deuterium affords greater metabolic stability (for example, increased in vivo half-life or reduced dosage requirements).

[0077] In embodiments, the compounds described herein are labeled by other means, including, but not limited to, the use of chromophores or fluorescent moieties, bioluminescent labels, or chemiluminescent labels.

[0078] In embodiments, the compounds described herein, and other related compounds having different substituents are synthesized using techniques and materials described herein and as described, for example, in Fieser and Fieser's Reagents for Organic Synthesis, Volumes 1-17 (John Wiley and Sons, 1991); Rodd's Chemistry of Carbon Compounds, Volumes 1-5 and Supplementals (Elsevier Science Publishers, 1989); Organic Reactions, Volumes 1-40 (John Wiley and Sons, 1991), Larock's Comprehensive Organic Transformations (VCH Publishers Inc., 1989), March, Advanced Organic Chemistry 4.sup.th Ed., (Wiley 1992); Carey and Sundberg, Advanced Organic Chemistry 4th Ed., Vols. A and B (Plenum 2000, 2001), and Green and Wuts, Protective Groups in Organic Synthesis 3rd Ed., (Wiley 1999) (all of which are incorporated by reference for such disclosure). General methods for the preparation of compound as described herein are modified by the use of appropriate reagents and conditions, for the introduction of the various moieties found in the formula as provided herein. See also Sergey I. Filimonov, Mikhail K. Korsakov, Dmitry V. Kravchenko, Mikhail V. Dorogov, Sergey E. Tkachenko, Alexandre V. Ivachtchenko, Convenient Synthesis of Novel 5-Substituted 3-Methylisoxazole-4-sulfonamides, J. Heterocyclic Chem., 43, 663 (2006)(e.g., see the left hand column of page 664, and Scheme 1, step iii shown therein and organic synthesis described therein); Hewings, Fedorov, Filippakopoulos, Martin, Picaud, Tumber, Wells, Olcina, Freeman, Gill, Ritchie, Sheppard, Russell, Hammond, Knapp, Brennan, and Conway, Optimization of 3,5-Dimethylisoxazole Derivatives as Potent Bromodomain Ligands, Journal of Medicinal Chemistry 2013 56 (8), 3217-3227; and A. Alberola, A. M. Gonzalez, M. A. Laguna, F. J. Pulido, Synthesis of 4-Functionalized 2-Isoxazolines by Reduction of the Isoxazole Ring with Complex Metal Hydrides, Synthesis 1983; 1983(5): 413-414 DOI: 10.1055/s-1983-30361 (all of which are entirely incorporated by reference).

[0079] Compounds described herein are synthesized using any suitable procedures starting from compounds that are available from commercial sources or are prepared using procedures described herein.

[0080] In embodiments, the one or more compounds of the present disclosure, or pharmaceutically acceptable salts or solvates thereof of the present disclosure are suitable for use in treating, ameliorating, or preventing diseases, disorders and conditions involving cancer. In embodiments, compositions are provided that are pharmaceutically acceptable, and/or may cross the blood/brain barrier. In embodiments, compositions of the present disclosure target, bind, and/or selectively bind to PP5 to minimize, alleviate or end downstream deleterious aspects associated therewith and/or induce an extrinsic apoptotic pathway. Non-limiting examples of diseases, disorders and conditions involving cancer include those described herein above such as sarcomas and the like.

[0081] Provided herein are methods of treating a medical condition involving cancer in a subject in need thereof by administering to the subject an effective amount of a polycyclic compound of the present disclosure such as those identified above and in the Figures. In embodiments, the medical condition involving cancer include one or more renal cell carcinomas (RCC) such as clear cell (ccRCC), papillary, chromophobe, collecting duct, or otherwise unclassified renal cell carcinoma. Other kidney cancers may include transitional cell carcinoma, Wilms' tumor, or renal sarcoma. In embodiments, conditions for treatment include metastasis.

[0082] In some embodiments, the effective amount is an amount sufficient to block one or more harmful effects of a nonfunctional extrinsic apoptotic pathway in one or more cancer cells. In embodiment, the effective amount is an amount sufficient to enable a nonfunctional extrinsic apoptotic pathway in a cancer cell or tumor. In embodiments, pharmaceutical agents of the present disclosure are provided in a therapeutically acceptable amount, in a form characterized as pharmaceutically acceptable.

[0083] In embodiments, the present disclosure includes a method of inhibiting cancer cells or a tumor by targeting PP5 in a subject in need thereof by administering to the subject a therapeutically effective amount of a polycyclic compound of the present disclosure such as those shown above and in the accompanying figures, or a pharmaceutically acceptable salt or solvate thereof.

[0084] In embodiments, the compositions of the present disclosure are suitable for use in a pharmaceutically acceptable formulation. In embodiments, the present disclosure includes a pharmaceutical formulation including a polycyclic compound of the present disclosure, or a pharmaceutically acceptable salt, and a pharmaceutically acceptable carrier. In embodiments, a polycyclic compound of the present disclosure may be referred to as an active compound. Pharmaceutical formulations including the active compounds also are provided herein. These pharmaceutical formulations include active compounds as described herein, in a pharmaceutically acceptable carrier. Pharmaceutical formulations can be prepared for oral, intravenous, or aerosol administration. Also, the presently disclosed subject matter provides such active compounds that have been lyophilized and that can be reconstituted to form pharmaceutically acceptable formulations for administration, for example, as by intravenous or intramuscular injection. In embodiments, a therapeutically effective dosage of any specific active compound, the use of which is within the scope of embodiments described herein, will vary somewhat from compound to compound, and patient to patient, and will depend upon the condition of the patient and the route of delivery.

[0085] Administration/Dosage/Formulations: In embodiments, provided herein is a pharmaceutical composition including at least one polycyclic compound of the present disclosure, or a pharmaceutically acceptable salt or solvate thereof. Actual dosage levels of the active ingredients in the pharmaceutical compositions of this disclosure may be varied to obtain an amount of the active ingredient that is effective to achieve the desired therapeutic response for a particular patient, composition, and mode of administration, without being toxic to the patient.

[0086] In embodiments, the selected dosage level will depend upon a variety of factors including the activity of the particular compound employed, the time of administration, the rate of excretion of the compound, the duration of the treatment, other drugs, compounds or materials used in combination with the compound, the age, sex, weight, condition, general health and prior medical history of the patient being treated, and like factors well, known in the medical arts.

[0087] A medical doctor, e.g., physician or veterinarian, having ordinary skill in the art may readily determine and prescribe the effective amount of the pharmaceutical composition required. For example, the physician or veterinarian could begin administration of the pharmaceutical composition to dose the polycyclic compound of the present disclosure, at levels lower than that required to achieve the desired therapeutic effect and gradually increase the dosage until the desired effect is achieved.

[0088] In some embodiments, the compounds and salts disclosed herein may be administered as a solvate in a continuous manner. For example, a single dose may be administered to a subject as a solvate (e.g., intravenously or a delayed release capsule) for an administration period of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 30, 45, or 60 minutes. In some embodiments, a single dose may be administered to a subject as a solvate for an administration period of 1-5, 5-10, 10-15, 15-30, 30-45, or 45-60 minutes. In some embodiments, a single dose may be administered to a subject as a solvate for an administration period of 1-60 minutes. In some embodiments a single dose may be administered to a subject for an administration period of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 24, 36, 48, 60, or 72 hours. In some embodiments a single dose may be administered to a subject for an administration period of 1-6, 6-12, 12-24, 24-48, 48-60, 60-72 hours. In some embodiments a single dose may be administered to a subject for an administration period of 1-72 hours.

[0089] In some embodiments, multiple doses of the compounds and salts described herein may be administered to a subject in a pulsatile or intermittent manner. As used herein the term pulsatile dose regimen or intermittent dose regimen refers to a dose administration regimen which includes at least two dosing cycles. Each subsequent dosing cycle is separated by a rest period from the preceding dosing cycle.

[0090] In particular embodiments, it is especially advantageous to formulate the compound in dosage unit form for ease of administration and uniformity of dosage. Dosage unit form as used herein refers to physically discrete units suited as unitary dosages for the patients to be treated, each unit containing a predetermined quantity of the polycyclic compound of the present disclosure calculated to produce the desired therapeutic effect in association with the required pharmaceutical vehicle. The dosage unit forms of the disclosure are dictated by and directly dependent on (a) the unique characteristics of the polycyclic compound of the present disclosure and the particular therapeutic effect to be achieved, and (b) the limitations inherent in the art of compounding/formulating such a polycyclic compound of the present disclosure for treatment in a patient.

[0091] In one embodiment, the compounds of the disclosure are formulated using one or more pharmaceutically acceptable excipients or carriers. In one embodiment, the pharmaceutical compositions of the invention comprise a therapeutically effective amount of a polycyclic compound of the present disclosure, and a pharmaceutically acceptable carrier.

[0092] Routes of administration of any of the compositions of the disclosure include oral, nasal, rectal, intravaginal, parenteral, buccal, sublingual or topical. The compounds for use in the disclosure may be formulated for administration by any suitable route, such as for oral or parenteral, for example, transdermal, transmucosal (e.g., sublingual, lingual, (trans)buccal, (trans)urethral, vaginal (e.g., trans- and perivaginally), (intra)nasal and (trans)rectal), intravesical, intrapulmonary, intraduodenal, intragastrical, intrathecal, subcutaneous, intramuscular, intradermal, intra-arterial, intravenous, intrabronchial, inhalation, and topical administration.

[0093] Suitable compositions and dosage forms include, for example, tablets, capsules, caplets, pills, gel caps, troches, dispersions, suspensions, solutions, syrups, granules, beads, transdermal patches, gels, powders, pellets, magmas, lozenges, creams, pastes, plasters, lotions, discs, suppositories, liquid sprays for nasal or oral administration, dry powder or aerosolized formulations for inhalation, compositions and formulations for intravesical administration and the like. It should be understood that the formulations and compositions that would be useful in the present disclosure are not limited to the particular formulations and compositions that are described herein.

[0094] For oral application, particularly suitable are tablets, dragees, liquids, drops, suppositories, or capsules, caplets and gelcaps. The compositions intended for oral use may be prepared according to any method known in the art and such compositions may contain one or more agents selected from the group consisting of inert, non-toxic pharmaceutically excipients that are suitable for the manufacture of tablets. Such excipients include, for example an inert diluent such as lactose; granulating and disintegrating agents such as cornstarch; binding agents such as starch; and lubricating agents such as magnesium stearate. The tablets may be uncoated or they may be coated by known techniques for elegance or to delay the release of the active ingredients. Formulations for oral use may also be presented as hard gelatin capsules wherein the active ingredient is mixed with an inert diluent.

[0095] For parenteral administration, the polycyclic compound of the present disclosure may be formulated for injection or infusion, for example, intravenous, intramuscular or subcutaneous injection or infusion, or for administration in a bolus dose or continuous infusion. Suspensions, solutions or emulsions in an oily or aqueous vehicle, optionally containing other formulatory agents such as suspending, stabilizing or dispersing agents may be used.

[0096] In some embodiments, the present disclosure includes a method of treating cancer, including: a) providing i) a subject with cancer, ii) one or more protein phosphatase-5 inhibitors, and b) treating said subject with the one or more PP5 inhibitors. In embodiments, treating includes administering a therapeutically effective amount of one or more protein phosphatase-5 inhibitors to the subject. In embodiments, the cancer is renal cancer such as renal cell carcinoma. In embodiments, the one or more protein phosphatase-5 inhibitors are characterized as a PP5 specific inhibitor. In embodiments, the one or more protein phosphatase-5 inhibitors are selected from the group consisting of:

##STR00021## ##STR00022## ##STR00023## ##STR00024## ##STR00025##

or a pharmaceutically acceptable salt thereof, and combinations thereof. In some embodiments, treating the subject with the one or more protein phosphatase-5 inhibitors is sequential. In embodiments, treating the subject with the one or more protein phosphatase-5 inhibitors is simultaneous. In embodiments, treating with the one or more protein phosphatase-5 inhibitors results in reduced proliferation of at least some of the cancer cells within said subject.

[0097] In some embodiments, the present disclosure includes a method of treating cancer, including a) providing i) a subject with cancer and one or more compounds of: selected from the group consisting of

##STR00026##