EP1986619A2 - Use of phosphatases to treat tumors overexpressing n-cor - Google Patents
Use of phosphatases to treat tumors overexpressing n-corInfo
- Publication number
- EP1986619A2 EP1986619A2 EP07763614A EP07763614A EP1986619A2 EP 1986619 A2 EP1986619 A2 EP 1986619A2 EP 07763614 A EP07763614 A EP 07763614A EP 07763614 A EP07763614 A EP 07763614A EP 1986619 A2 EP1986619 A2 EP 1986619A2
- Authority
- EP
- European Patent Office
- Prior art keywords
- cor
- cells
- ligand
- patient
- tumor
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Withdrawn
Links
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
- A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
- A61K31/519—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim ortho- or peri-condensed with heterocyclic rings
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
Definitions
- GBM glioblastoma multiforme
- GBM is the most common primary brain tumor.
- GBM is also the most malignant primary brain tumor.
- GBM' s malignancy typically remains in the cerebral hemispheres of the brain; however, glioblastomas can form in the brainstem, the cerebellum and the spinal cord. GBM does not usually spread to other parts of the body.
- GBM tumors form from the supportive or glial tissue of the brain. GBM tumor cells look very different from normal brain cells. GBM cells are poorly differentiated, neoplastic astrocytes. GBM tumors are characterized by molecular lesions, cellular pleomorphism, mitotic figures, and multinucleated giant cells. (U.S. Patent Publication No. 2005/0203082, Hsu et al.) The World Health Organization classifies GBM as having 3 or 4 of the following histologic criteria: (1) nuclear atypia,
- GBM tumors may develop from much less malignant precursor tumors, called astrocytomas (secondary GBMs), or it may form de novp, with no evidence of a precursor tumor (primary GBM) .
- the subject invention provides novel methods of treating GBM. It also provides novel methods of diagnosing and screening for this deadly disease. Summary of the Invention
- the invention provides a method of treating a patient suffering from a tumor overexpressing N-CoR comprising administering to the patient one or more phosphatase ligand, alone or in combination with one or more retinoid receptor ligand, or one or more histone deacetylase ligand, or both, in each case in an amount effective to treat the patient.
- This invention also provides a method of inhibiting growth of a tumor overexpressing N-CoR in a patient, comprising administering to the patient one or more phosphatase ligand, alone or in combination with one or more retinoid receptor ligand, or one or more histone deacetylase ligand, or both, in each case in amounts effective to affect N-CoR so as to induce differentiation of cells of the tumor overexpressing N-CoR and inhibit growth of the tumor in the patient.
- This invention further provides a method of identifying a compound or a mixture of compounds capable of inducing differentiation or inhibiting proliferation of cells of a tumor overexpressing N-CoR, comprising the steps of (a) culturing a first population of the specified human cells in the absence of the compound or the mixture of compounds in both serum and serum free conditions; (b) separately culturing a second population of such human cells in the presence of the compound or the mixture of compounds; (c) comparing the rate of growth of the cultured human cells in step (a) with the rate of growth of the cultured human cells in step (b) ; (d) identifying the compound or the mixture of compounds which inhibited, or reduced the rate of, growth of the cultured human cells in step (b) as compared to the rate of growth of the cultured human cells in step (a); and (e) measuring the level of N-CoR in the cytoplasm and in the nucleus of the cultured human cells from step (b) whose growth was inhibited or whose rate of growth was reduced in the presence
- This invention also provides a method of determining the likelihood of successfully treating a subject suffering from a tumor overexpressing N-CoR, comprising the steps of (a) obtaining a sample from the subject containing cells of a tumor overexpressing N-CoR; and (b) measuring the level of N- CoR in the cytoplasm and in the nucleus of cells in the sample so obtained, wherein the presence in the sample of an increased level of N-CoR in the nucleus of the cells indicates that there is a greater likelihood of successfully treating the subject.
- This invention further provides a method of assessing the likelihood that a patient is suffering from a tumor overexpressing N-CoR, comprising the steps of (a) obtaining a serum sample from the subject; and (b) measuring the level of N-CoR in the serum sample so obtained, wherein the presence in the serum sample of increased levels of N-CoR relative "to a normal reference standard indicates that the patient is likely suffering from a tumor overexpressing N-CoR.
- This invention still further provides a method of assessing the likelihood that a patient previously suffering from and treated for a tumor overexpressing N-CoR has suffered a recurrence of such tumor, comprising the steps of (a) obtaining a serum sample from the subject; and (b) measuring the level of N-CoR in the serum sample so obtained, wherein the presence in the serum sample of increased levels of N-CoR relative to a previous level of N-CoR indicates that the patient is likely suffering from a recurrence of a tumor overexpressing N-CoR.
- This invention provides a method of treating a patient suffering from glioblastoma multiforme, comprising administering to the patient one or more phosphatase ligand, alone or in combination with one or more retinoid receptor ligand, or one or more histone deacetylase ligand, or both, in each case in amounts effective to treat the patient.
- This invention also provides a method of inhibiting growth of a tumor in a patient suffering from glioblastoma multiforme, comprising administering to the patient one or more phosphatase ligand, alone or in combination with one or more retinoid receptor ligand, or one or more histone deacetylase ligand, or both, in each case in amounts effective to affect N-CoR so as to induce differentiation of glioblastoma multiforme tumor cells and inhibit growth of the tumor in the patient.
- This invention further provides a method of identifying a compound or a mixture of compounds capable of inducing differentiation or inhibiting proliferation of glioblastoma multiforme tumor cells, comprising the steps of (a) culturing a first population of human brain cells in the absence of the compound or the mixture of compounds in both serum and serum free conditions; (b) separately culturing a second population of such human brain cells in the presence of the compound or the mixture of compounds; (c) comparing the rate of growth of the cultured human brain cells in step (a) with the rate of growth of the cultured human brain cells in step (b) ; (d) identifying the compound or the mixture of compounds which inhibited, or reduced the rate of, growth of the cultured human brain cells in step (b) as compared to the rate of growth of the cultured human brain cells in step (a); and (e) measuring the level of N-CoR and the level of a glioblastoma multiforme lineage marker in the cytoplasm and in the nucleus of the cultured human
- This invention also provides a method of determining the likelihood of successfully treating a subject suffering from glioblastoma multiforme, comprising the steps of (a) obtaining a sample from the subject containing glioblastoma multiforme cells; and (b) measuring the level of each of N-CoR and a glioblastoma multiforme lineage marker in the cytoplasm and in the nucleus of cells in the sample so obtained, wherein the presence in the sample of an increased level of N-CoR and a low or undetectable level of glioblastoma multiforme lineage marker in the cytoplasm of the cells indicates that there is a greater likelihood of successfully treating the subject.
- This invention still further provides a method of assessing the likelihood that a patient is suffering from glioblastoma multiforme, comprising the steps of (a) obtaining a sample ' of cerebrospinal fluid and/or tumor cells or serum from the subject; and (b) measuring the level of N-CoR in the cerebrospinal fluid and/or the cells or serum in the sample so obtained, wherein the presence in the sample of increased levels of N-CoR in the cerebrospinal fluid relative to a normal reference standard indicates that the patient is likely suffering from glioblastoma multiforme. If N-CoR is increased in the serum but not in the cerebral spinal fluid this would indicate that the patient is likely suffering from a tumor overexpressing N-CoR but not necessarily a glioblastoma multiforme.
- this invention provides a method of assessing the likelihood that a patient previously suffering from and treated for glioblastoma multiforme has suffered a recurrence of glioblastoma multiforme, comprising the steps of (a) obtaining a sample of cerebrospinal fluid and/or tumor cells or serum from the subject; and (b) measuring the level of N- CoR in the cerebrospinal fluid and/or in the cells or serum in the sample so obtained, wherein the presence in the sample of increased levels of N-CoR in the cerebrospinal fluid or serum relative to the amount of N-CoR previously in the cerebral spinal fluid indicates that the patient is likely suffering from a recurrence of glioblastoma multiforme.
- Figure IA Differential expression of N-CoR in normal and GBM brain tissue.
- Total proteomic analysis of microdissected normal glial tissue (white matter) compared with GBM was performed by two dimensional gel electrophoresis (2-DGE) .
- the highlighted region which is magnified on the right panels shows a consistent protein pattern in normal glial and GBM and unique expression of N-CoR in GBM.
- Protein identification was performed by liquid chromatography-mass spectrometry.
- FIG. 1B Expression of N-CoR in GBM by immunohistochemistry: N-CoR protein is present in both nucleus and cytoplasm in GBM. Right panel: arrows point to N-CoR staining (circled) in nucleus (right) and cytoplasm (left) . Left panel: no N-CoR is seen in normal tissue.
- FIG. 1C Expression of N-CoR in GBM by Western blot analysis: N-CoR is present in GBMs on lanes 2-4 (molecular weight 270 k.Da) . N-CoR is absent in normal white matter (lane 1) . ⁇ -actin was used • as internal positive quantitative control .
- N-CoR Neuronal differentiation
- N-CoR and GFAP immunolabeling in GBM.
- Cell on right demonstrating nuclear N-CoR localization shows no cytoplasmic GFAP.
- Cell on left with absent N-CoR labeling shows cytoplasmic expression of GFAP.
- Figure IE Co-expression of nuclear localization of N-CoR and cytoplasmic expression of CD133 in GBM primary culture: Immunolabeling of nuclear localization of N-CoR and cytoplasmic CD133 are present in the same GBM cells.
- FIG. 2A GFAP expression by CNTF-treated BTSC: GFAP expression was induced in glioma stem cells by treatment with CNTF and detected by 2-DGE and LCMS. Arrow points to the GFAP spot (circled) .
- Figure 2B Cytoplasmic N-CoR fraction increased by CNTF treatment of BTSC. Cytoplasmic level of N-CoR expression in BTSC shows gradual increase from day 0 to day 7 upon CNTF treatment, ⁇ -actin is shown as quantitative internal control.
- Figure 2C Logarithmic growth curve of gliomal cell line, U343 MG-A, treated with retinoic acid (RA) , okadaic acid (OA) , combination of retinoic acid and okadaic acid (RA/0A) , and control (NC) for 16 days: Individual treatment with retinoic acid and okadaic acid show a modest inhibition of growth. Combination of retinoic acid and okadaic acid shows synergistic reduction in cell growth. Error bars indicate 1 SD.
- Figure 4A Logarithmic curve of gliomal cell line -U373 treated with all-trans retinoic acid (ATRA) . Increasing dosage shows a modest inhibition of growth. Error bars indicate SD.
- Figure 4B Inhibition of gliomal cell line U373 treated with endothal (End) and compound LB-I with and without all- trans Retinoic acid (ATRA) for 7 days. Individual treatment with endothal and compound LB-I shows modest inhibition of growth. Combination of End or compound LB-I with ATRA shows synergistic reduction in cell growth. Error bars indicate SD.
- Figure 4C Inhibition of growth of gliomal cell line U373 by endothal (End) with and without 13-cis Retinoic Acid (cis- RA) .
- End endothal
- cis- RA 13-cis Retinoic Acid
- Individual treatment with End and cis-RA show a modest inhibition of growth.
- Combination of End and cis-RA show a synergistic reduction in cell growth. Error bars indicate SD.
- FIG. 5A Inhibition of growth of gliomal cell line U373 with Valproic Acid (VaI) . Increasing doses of VaI (mM) shows a greater inhibition of cell growth. Error bars indicate SD.
- FIG. 5B Inhibition of growth of gliomal cell line U373 by Trichostatin A (TSA) .
- TSA Trichostatin A
- Increasing doses of TSA show a greater inhibition of cell growth. Error bars indicate SD.
- FIG. 6A Inhibition of growth of kidney cancer cell line, UMRC by endothal thioanhydride (ET) , endothal (End) , all-trans Retinoic Acid (ATRA) , Trichostatin A (TSA) and norcantharidin (nor-Can) for 7 days. Error bars indicate SD
- FIG. 6B Inhibition of growth of gliomal cell line U373 by endothal thioanhydride (ET) , endothal (End) , all-trans Retinoic Acid (ATRA), Trichostatin A (TSA) and norcantharidin ' (nor-Can) for 7 days. Individual treatment with endothal thioanhydride showed the greatest inhibition of growth. Error bars indicate SD.
- FIG. 6C Inhibition of growth of breast cancer cell line, MCF-7 by Inhibition of UMRC by endothal thioanhydride (ET) , endothal (End) , all-trans Retinoic Acid (ATRA) , Trichostatin A (TSA) and norcantharidin (nor-Can) for 7 days.
- EMT endothal thioanhydride
- ATRA all-trans Retinoic Acid
- TSA Trichostatin A
- norcantharidin norcantharidin
- administering an agent may be performed using any of the various methods or delivery systems well known to those skilled in the art.
- the administering can be performed, for example, orally, parenterally, intraperitoneally, intravenously, intraarterially, transdermally, sublingually, intramuscularly, rectally, transbuccally, intranasally, liposomally, via inhalation, vaginally, intraoccularly, via local delivery, subcutaneously, intraadiposally, intraarticularly, intrathecally, into a cerebral ventricle, intraventicularly, intratumorally, into cerebral parenchyma or intraparenchchymally.
- compositions in accordance with the invention may be used but are only representative of the many possible systems envisioned for administering compositions in accordance with the invention .
- Injectable drug delivery systems include solutions, suspensions, gels, microspheres and polymeric injectables, and can comprise excipients such as solubility-altering agents (e.g., ethanol, propylene glycol and sucrose) and polymers (e.g., polycaprylactones and PLGA' s).
- solubility-altering agents e.g., ethanol, propylene glycol and sucrose
- polymers e.g., polycaprylactones and PLGA' s.
- Implantable systems include rods and discs, and can contain excipients such as PLGA and polycaprylactone.
- Oral delivery systems include tablets and capsules. These can contain excipients such as binders (e.g., hydroxypropylmethylcellulose, polyvinyl pyrilodone, other cellulosic materials and starch), diluents (e.g., lactose and other sugars, starch, dicalcium phosphate and cellulosic materials), disintegrating agents (e.g., starch polymers and cellulosic materials) and lubricating agents (e.g., stearates and talc) .
- binders e.g., hydroxypropylmethylcellulose, polyvinyl pyrilodone, other cellulosic materials and starch
- diluents e.g., lactose and other sugars, starch, dicalcium phosphate and cellulosic materials
- Transmucosal delivery systems include patches, tablets, suppositories, pessaries, gels and creams, and can contain excipients such as solubilizers and enhancers (e.g., propylene glycol, bile salts and amino acids), and other vehicles (e.g., polyethylene glycol, fatty acid esters and derivatives, and hydrophilic polymers such as hydroxypropylmethylcellulose and hyaluronic acid) .
- solubilizers and enhancers e.g., propylene glycol, bile salts and amino acids
- other vehicles e.g., polyethylene glycol, fatty acid esters and derivatives, and hydrophilic polymers such as hydroxypropylmethylcellulose and hyaluronic acid
- Dermal delivery systems include, for example, aqueous and nonaqueous gels, creams, multiple emulsions, microemulsions, liposomes, ointments, aqueous and nonaqueous solutions, lotions, aerosols, hydrocarbon bases and powders, and can contain excipients such as solubilizers, permeation enhancers (e.g., fatty acids, fatty acid esters, fatty alcohols and amino acids), and hydrophilic polymers (e.g., polycarbophil and polyvinylpyrolidone) .
- the pharmaceutically acceptable carrier is a liposome or a transdermal enhancer.
- Solutions, suspensions and powders for reconstitutable delivery systems include vehicles such as suspending agents (e.g., gums, zanthans, cellulosics and sugars), humectants (e.g., sorbitol), solubilizers (e.g., ethanol, water, PEG and propylene glycol), surfactants (e.g., sodium lauryl sulfate, Spans, Tweens, and cetyl pyridine), preservatives and antioxidants (e.g., parabens, vitamins E and C, and ascorbic acid) , anti-caking agents, coating agents, and chelating agents (e.g., EDTA).
- suspending agents e.g., gums, zanthans, cellulosics and sugars
- humectants e.g., sorbitol
- solubilizers e.g., ethanol, water, PEG and propylene glycol
- terapéuticaally effective amount means an amount sufficient to treat a subject afflicted with a disease (e.g. glioblastoma multiforme) or to alleviate a symptom or a complication associated with the disease.
- a disease e.g. glioblastoma multiforme
- treating means slowing, stopping or reversing the progression of a disease, particularly glioblastoma multiforme.
- N-CoR nuclear co-receptor
- the nuclear receptor co-repressor (N-CoR) of the subject invention may be any molecule that binds to the ligand binding domain of the DNA-bound thyroid hormone receptor ⁇ T 3 R) and retinoic acid receptor (RAR).
- N-CoR nuclear receptor co-repressor
- Examples of tumors that overexpress N-CoR may include glioblastoma multiforme, breast cancer (Myers et al . ) , colorectal cancer (Giannini and Cavallini) , small cell lung cancer (Waters et al.) and ovarian cancer (Hdressesky et al.).
- the invention provides a method of treating a patient suffering from a tumor overexpressing N-CoR comprising administering to the patient one or more phosphatase ligand, alone or in combination with one or more retinoid receptor ligand, or one or more histone deacetylase ligand, or both, in each case in an amount effective to treat the patient.
- the phosphatase ligand may be selected from the group consisting of 1-nor-okadaone, antimonyl tartrate, bioallethrin, calcineurin, cantharidic acid, cantharidin, calyculin, cypermethrin, DARPP-32, deamidine, deltamethrin, diaminopyrroloquinazolines, endothal, endothal thioanhydride, fenvalerate, ' fostriecin, imidazoles, ketoconazole, L-4- bromotetramisole, levamisole, microcystin LA, microcystin LR, microcystin LW, microcystin RR, molybdate salts, okadaic acid, okadol, norcantharidin, pentamidine, pentavalent antimonials, permethrin, phenylarsine oxide, phloridzin, protein phosphat
- the histone deacetylase ligand may be an inhibitor, e.g. the histone deacetylase inhibitor HDAC-3 (histone deacetylase-3) .
- the histone deacetylase ligand may also be selected from the group consisting of 2-amino-8- oxo-9, 10-epoxy-decanoyl, 3- (4-aroyl-lH-pyrrol-2-yl) -N-hydroxy- 2-propenamide, APHA Compound 8, apicidin, arginine butyrate, butyric acid, depsipeptide, depudecin, HDAC-3, m- carboxycinnamic acid bis-hydroxamide, N- (2-aminophenyl) -4- [N- (pyridin-3-ylmethoxycarbonyl) aminomethyl] benzamide, MS 275, oxamfiatin, phenylbutyrate, pyroxamide, scriptaid, sirtin
- Histones are groups of proteins which associate with DNA in eukaryotic cells to form compacted structures called chromatin. This compaction allows an enormous amount of DNA to be located within the nucleus of a eukaryotic cell, but the compact structure of chromatin restricts the access of transcription factors to the DNA. Acetylation of the histones decreases the compaction of the chromatin allowing transcription factors to bind to the DNA.
- HDACs histone deacetylases
- This invention also provides a method of inhibiting growth of a tumor overexpressing N-CoR in a patient, comprising administering to the patient one or more phosphatase ligand, alone or in combination with one or more retinoid receptor ligand, one or more histone deacetylase ligand, or both, in each case in amounts effective to affect N-CoR so as to thereby induce differentiation of cells of the tumor overexpressing N-CoR and inhibit growth of the tumor in the patient .
- This invention further provides a method of identifying a compound or a mixture of compounds capable of inducing differentiation or inhibiting proliferation of cells of a tumor overexpressing N-CoR, comprising the steps of (a) culturing a first population of the specified human cells in the absence of the compound or the mixture of compounds in both serum and serum free conditions; (b) separately culturing a second population of such human cells in the presence of the compound or the mixture of compounds; (c) comparing the rate of growth of the cultured human cells in step (a) with the rate of growth of the cultured human cells in step (b) ; (d) identifying the compound or the mixture of compounds which inhibited, or reduced the rate of, growth of the cultured human cells in step (b) as compared to the irate of growth of the cultured human cells in step (a); and (e) measuring the level of N-CoR in the cytoplasm and in the nucleus of the cultured human cells from step (b) whose growth was inhibited or whose rate of growth was reduced in
- the level of N-CoR in the cytoplasm and in the nucleus may be measured by either indirect immunofluorescence microscopy, direct immunofluorescence microscopy, FACS, or other methods for detecting and measuring amounts of specific proteins in tissues including assessment of the amounts of proteins in the nucleus versus the cytoplasm and in cell lysates, or a combination thereof.
- N-CoR is expressed in the nucleus of the undifferentiated tumor or stem cells and is only present in the cytoplasm at amounts detectable by i ⁇ ununochemistry and Western blotting when the cell undergoes differentiation. N-CoR " is not detectable by immunochemistry and Western blotting in either the nucleus or the cytoplasm of normal or fully differentiated cells.
- an assessment of the percentage of cells with N-CoR in the cytoplasm relative to the percentage of cells with N-CoR in the nucleus is representative of the ratio of more differentiated cells to less differentiated cells in a given tissue.
- tumors that overexpress N-CoR may include glioblastoma multiforme, breast cancer, colorectal cancer, small cell lung cancer and ovarian cancer.
- This invention also provides a method of determining the likelihood of successfully treating a subject suffering from a tumor overexpressing N-CoR, comprising the steps of (a) obtaining a sample from the subject containing cells of a tumor overexpressing N-CoR; and (b) measuring the level of N- CoR in the cytoplasm and in the nucleus of cells in the sample so obtained, wherein the presence in the sample of an increased level of N-CoR in the nucleus of the cells indicates that there is a greater likelihood of successfully treating the subject.
- the level of N-CoR in the cytoplasm and in the nucleus may be measured by either indirect immunofluorescence microscopy, direct immunofluorescence microscopy, FACS, or other methods for detecting and measuring amounts of specific proteins in tissues including assessment of the amounts of proteins in the nucleus versus the cytoplasm and in cell lysates, or a combination thereof.
- N-CoR is expressed in the nucleus of the undifferentiated tumor or stem cells and is only present in the cytoplasm at amounts detectable by immunochemistry and Western blotting when the cell undergoes differentiation. N-CoR is not detectable by immunochemistry and Western blotting in either the nucleus or the cytoplasm of normal or fully differentiated cells.
- an assessment of the percentage of cells with N-CoR in the cytoplasm relative to the percentage of cells with N-CoR in the nucleus is representative of the ratio of more differentiated cells to less differentiated cells in a given tissue.
- tumors that overexpress N-CoR may include glioblastoma multiforme, breast cancer, colorectal cancer, small cell lung cancer and ovarian cancer.
- This invention further provides a method of assessing the likelihood that a patient is suffering from a tumor overexpressing N-CoR, comprising the steps of (a) obtaining a serum sample from the subject; and (b) measuring the level of N-CoR in the serum sample so obtained, wherein the presence in the serum sample of increased levels of N-CoR relative to a normal reference standard indicates that the patient is likely suffering from a tumor overexpressing N-CoR.
- tumors that overexpress N-CoR may include glioblastoma multiforme, breast cancer, colorectal cancer / small cell lung cancer and ovarian cancer.
- This invention still further provides a method of assessing the likelihood that a patient previously suffering from and treated for a tumor overexpressing N-CoR has suffered a recurrence of such tumor, comprising the steps of (a) obtaining a serum sample from the subject; and (b) measuring the level of N-CoR in the serum sample so obtained, wherein the presence in the serum sample of increased levels of N-CoR relative to a previously lower level indicates that the patient is likely suffering from a recurrence of a tumor overexpressing N-CoR.
- tumors that overexpress N-CoR may include glioblastoma multiforme, breast cancer, colorectal cancer, small cell lung cancer and ovarian cancer.
- This invention provides a method of treating a patient suffering from glioblastoma multiforme, comprising administering to the patient one or more phosphatase ligand, alone or in combination with one or more retinoid receptor ligand, or one or more histone deacetylase ligand, or both, in each case in amounts effective to treat the patient.
- the phosphatase ligand may be selected from the group consisting of 1-nor-okadaone, antimonyl tartrate, bioallethrin, calcineurin, cantharidic acid, cantharidin, calyculin, cypermethrin, DARPP-32, deamidine, deltamethrin, diaminopyrroloquinazolines, endothal, endothal thioanhydride, fenvalerate, fostriecin, imidazoles, ketoconazole, L-4- bromotetramisole, levamisole, 1-p-bromotetramisole, d-p- bromotetramisole, p-hydroxylevamisole, microcystin LA, microcystin LR, microcystin LW, microcystin RR, molybdate salts, okadaic acid, okadol, norcantharidin, pentamidine, penta
- the phosphatase ligand is a protein phosphatase inhibitor, such as endothal thioanhydride, endothal, norcantharidin or okadaic acid.
- the protein phosphatases of the subject application can be tyrosine-specific, serine/threonine-specific, dual-specificity phosphatases, alkaline phosphatases such as levamisole, and acid phosphatases.
- the retinoid receptor ligand may be a retinoid, such as a retinoic acid, e.g. cis retinoic acid or trans retinoic acid.
- a retinoic acid e.g. cis retinoic acid or trans retinoic acid.
- the cis retinoic acid may be 13- cis retinoic acid and the trans retinoic acid may be all-trans retinoic acid.
- the retinoid receptor ligand may affect retinoid receptor activity but not thyroid hormone receptor activity; alternatively or additionally the retinoid receptor ligand may inhibit N-CoR binding to the retinoid receptor but not N-CoR binding to the thyroid hormone receptor.
- Retinoid receptor ligands used in the method of the invention include vitamin A (retinol) and all its natural and synthetic derivatives (retinoids) .
- the retinoid receptor ligand may be selected from the group consisting of b, g-selective 6- (5,6,7, 8-tetrahydro-5, 5,8, 8-tetramethyl-2-naphthalenyl) -2- naph-thalenecarboxylic acid (TTNN), Z-oxime of 6- (5, 6,7,8- tetrahydro-5, 5,8, 8-tetramethyl-2-naphthalenylcarbonyl) -2- naphthalenecarboxyli ⁇ acid (SR11254), 4- (5, 6, 7 , 8-tetrahydro- 5, 5, 8,8-tetramethyl-2-anthracenyl) benzoic acid (TTAB), 4-[l- (5, 6, 7, 8-tetrahydro-5, 5,8, 8-tetramethyl-2-naphthalenyl) - cyclopropyl] benzoic acid (SR11246) , 4- [1- (5, 6, 7 , 8-
- the histone deacetylase ligand may be an inhibitor, e.g. the histone deacetylase inhibitor
- HDAC-3 histone deacetylase-3
- the histone deacetylase ligand may also be selected from the group consisting of 2-amino-8- oxo-9, 10-epoxy-decanoyl, 3- (4-aroyl-lH-pyrrol-2-yl) -N-hydroxy-
- HDAC histone deacetylase
- HDAC enzymes posttranslationally modify histones are groups of proteins which associate with DNA in eukaryotic cells to form compacted structures called chromatin. This compaction allows an enormous amount of DNA to be located within the nucleus of a eukaryotic cell, but the compact structure of chromatin restricts the access of transcription factors to the DNA. Acetylation of the histones decreases the compaction of the chromatin allowing transcription factors to bind to the DNA. Deacetylation, catalysed by histone deacetylases (HDACs) , increases the compaction of chromatin, thereby reducing transcription factor accessibility to DNA. Therefore, inhibitors of histone deacetylases prevent the compaction of chromatin, allowing transcription factors to bind to DNA and increase expression of the genes.
- HDACs histone deacetylases
- This invention also provides a method of inhibiting growth of a tumor in a patient suffering from glioblastoma multiforme, comprising administering to the patient one or more phosphatase ligand, alone or in combination with one or more retinoid receptor ligand, one or more histone deacetylase ligand, or both, in each case in amounts effective to affect N-CoR so as to thereby induce differentiation of glioblastoma multiforme tumor cells and inhibit growth of the tumor in the patient .
- N-CoR nuclear receptor co-repressor
- T 3 R DNA-bound thyroid hormone receptor
- RAR retinoic acid receptor
- This invention further provides a method of identifying a compound or a mixture of compounds capable of inducing differentiation or inhibiting proliferation of glioblastoma multiforme tumor cells, comprising the steps of (a) culturing a first population of human brain cells in the absence of the compound or the mixture of compounds in both serum and serum free conditions; (b) separately culturing a second population of such human brain cells in the presence of the compound or the mixture of compounds; (c) comparing the rate of growth of the cultured human brain cells in step (a) with the rate of growth of the cultured human brain cells in step (b) ; (d) identifying the compound or the mixture of compounds which inhibited, or reduced the rate of, growth of the cultured human brain cells in step (b) as compared to the rate of growth of the cultured human brain cells in step (a); and (e) measuring the level of N-CoR and the level of a glioblastoma multiforme lineage marker in the cytoplasm and in the nucleus of the cultured human
- step (a) wherein a decrease in the level of N-Cor and an increase in glioblastoma multiforme lineage marker indicate that the compound or the mixture of compounds is capable of inducing differentiation of glioblastoma multiforme tumor cells, so as to thereby identify the compound or the mixture of compounds.
- the glioblastoma multiforme lineage marker may be selected from the group consisting of GFAP, nestin, tujl, and CNPase.
- a glial fibrillary acidic protein (GFAP) useful in the subject invention is a 55 kDa cytosolic protein, a major structural component of astroglial filaments and the major intermediate filament protein in astrocytes.
- GFAP is specific to astrocytes of the brain.
- the level of N-CoR and the level of the glioblastoma multiforme lineage marker in the cytoplasm and in the nucleus may be measured by either indirect immunofluorescence microscopy, direct immunofluorescence microscopy, FACS, or other methods for detecting and measuring amounts of specific proteins in tissues including assessment of the amounts of proteins in the nucleus versus the cytoplasm and in cell lysates, or a combination thereof.
- N-CoR is expressed in the nucleus of the undifferentiated tumor or stem cells and is only present in the cytoplasm at amounts detectable by immunochemistry and Western blotting when the cell undergoes differentiation. N-CoR is not detectable by immunochemistry and Western blotting in either the nucleus or the ' cytoplasm of normal or fully differentiated cells .
- an assessment of the percentage of cells with N-CoR in the cytoplasm relative to the percentage of cells with N-CoR in the nucleus is representative of the ratio of more differentiated cells to less differentiated cells in a given tissue.
- the first population of human brain cells and the second population of human brain cells is selected from the group consisting of primary normal human brain cells, primary human brain stem cells, and primary glioblastoma multiforme stem cells.
- the first population of human brain cells and the second population of human brain cells may be the same or different, preferably the same and may be cells derived from any of the following cell lines: U343 MG-A, U251, U373, U87, A-172, LN-18, LN-229, M059J, M059K, and HS683.
- Cell line U343 MG-A is available from the University of California at San Francisco (UCSF) Brain Tumor Research Center Tissue Bank. (University of California, San Francisco, Health Sciences West building, San Francisco, California 94143-0520.) In addition, cell lines U343 and U87 are commercially available from EPO-GmbH, Robert-R ⁇ ssle-Str.10, 13092 Berlin- Buch, Germany.
- Cell line U251 is available from Division of Cancer Treatment and Diagnosis at National Cancer Institute Tumor Repository, The National Cancer Institute at Frederick Bldg. 1073, Frederick, Maryland 21702-1201.
- Cell lines A-172, LN-18, LN-229, M059J, M059K, and HS683 are available from the American Type Culture Collection (ATCC) , P.O. Box 1549, Manassas, Virginia, 20108, as ATCC No. CRL- 1620, ATCC No. CRL-2610, ATCC No. CRL-229, ATCC No. CRL-2365 and ATCC No. HTB-138, respectively.
- ATCC American Type Culture Collection
- Cell line U373 is available from the National Institute of Neurological Disease and Stroke, Building 31, 31 Center Drive, Bethesda, MD, 20892 and the National Institute of Health, Building 1, 1 Center Drive, Bethesda, Maryland, 20892.
- This invention also provides a method of determining the likelihood of successfully treating a subject suffering from glioblastoma multiforme, comprising the steps of (a) obtaining a sample from the subject containing glioblastoma multiforme cells; and (b) measuring the level of each of N-CoR and a glioblastoma multiforme lineage marker in the cytoplasm and in the nucleus of cells in the sample so obtained, wherein the presence in the sample of an increased level of N-CoR in the nucleus indicated that there is a greater likelihood of successfully treating the subject.
- the glioblastoma multiforme lineage marker may be selected from the group consisting of GFAP, nestin, tujl, and CNPase.
- the level of N-CoR and the level of the glioblastoma multiforme lineage marker in the cytoplasm and in the nucleus may be measured by either indirect immunofluorescence microscopy, direct immunofluorescence microscopy, FACS, or other methods for detecting and measuring amounts of specific proteins in tissues including assessment of the amounts of proteins in the nucleus versus the cytoplasm and in cell lysates, or a combination thereof.
- N-CoR is expressed in the nucleus of the undifferentiated tumor or stem cells and is only present in the cytoplasm at amounts detectable by immunochemistry and Western blotting when the cell undergoes differentiation. N-CoR is not detectable by immunochemistry and Western blotting in either the nucleus or the cytoplasm of normal or fully differentiated cells .
- an assessment of the percentage of cells with N-CoR in the cytoplasm relative to the percentage of cells with N-CoR in the nucleus is representative of the ratio of more differentiated cells to less differentiated cells in a given tissue.
- This invention also provides a method of assessing the likelihood that a patient is suffering from glioblastoma multiforme, comprising the steps of (a) obtaining a sample of cerebrospinal fluid and/or tumor cells from the subject; and (b) measuring the level of N-CoR in the cerebrospinal fluid and/or the cells in the sample so obtained, wherein the presence in the sample of increased levels of N-CoR in the cerebrospinal fluid relative to a normal reference standard indicates that the patient is likely suffering from glioblastoma multiforme. If N-CoR is increased in the serum but not in the cerebral spinal fluid this would indicate that the patient is likely suffering from a tumor overexpressing N- CoR but not necessarily a glioblastoma multiforme.
- This invention also provides a method of assessing the likelihood that a patient previously suffering from and treated for glioblastoma multiforme has suffered a recurrence of glioblastoma multiforme, comprising the steps of (a) obtaining a sample of cerebrospinal fluid and/or tumor cells from the subject; and (b) measuring the level of N-CoR in the cerebrospinal fluid and/or the cells in the sample so obtained, wherein the presence in the sample of increased levels of N-CoR in the cerebrospinal fluid relative to the previous levels of N-CoR post-treatment indicates that the patient is likely suffering from a recurrence of glioblastoma multiforme.
- This invention further provides a method of assessing the likelihood that a patient is suffering from a tumor overexpressing N-CoR, comprising the steps of (a) obtaining a serum sample from the subject; and ⁇ b) measuring the level of N-CoR in the serum sample so obtained, wherein the presence in the serum sample of increased levels of N-CoR relative to a normal reference standard indicates that the patient is likely suffering from a tumor overexpressing N-CoR.
- This invention still further provides a method of assessing the likelihood that a patient previously suffering from and treated for a tumor overexpressing N-CoR has suffered a recurrence of a tumor overexpressing N-CoR, comprising the steps of (a) obtaining a serum sample from the subject; and
- the invention provides a use of one or more phosphatase ligand, in an amount effective to treat a patient, alone or in combination with one or more retinoid receptor ligand, or one or more histone deacetylase ligand, or both, in each case in an amount effective to treat the patient, for the preparation of a medicament.
- the medicament comprises one or more phosphatase ligand alone, or for use with, one or more retinoid receptor ligand, or one or more histone deacetylase receptor ligand, or both.
- This invention also provides the use of a phosphatase ligand, in an amount effective to induce differentiation of cells of a tumor overexpressing N-CoR ⁇ and to inhibit the growth of the tumor in a patient, alone or in combination with one or more retinoid receptor ligand, one or more histone deacetylase ligand, or both, for the preparation of a medicament.
- the medicament comprises one or more phosphatase ligand alone, or for use with, one or more retinoid receptor ligand, or one or more histone deacetylase receptor ligand, or both.
- the uses of the invention herein encompass the enumerated phosphatase ligands, retinoid receptors and histone deacetylase receptor ligands enumerated above.
- the invention provides a product containing a phosphatase ligand in combination with one or more retinoid receptor ligand, one or more histone deacetylase ligand, or both, as a combined preparation for simultaneous, separate or sequential use in treating a tumor overexpressing N-CoR.
- proteomes of 7 GBM tissues and 7 normal brain tissues were compared using selective microdissection, two dimensional gel electrophoresis (2-DGE) and liquid chromatography-mass spectroscopy (LCMS) .
- GBM tissue was further tested by immunohistochemistry and Western blotting for the expression of nuclear receptor co- repressor (N-CoR) .
- N-CoR nuclear receptor co- repressor
- ⁇ -actin was used as internal positive quantitative control for the Western blotting.
- GFAP glial fibrillary acidic protein
- GBM cell lines were all cultured in DMEM with 10% FCS and high glucose DMEM/F12 with N2 supplement (serum-free) on poly-
- BTSC brain tumor stem cells isolated from GBM were treated with .ciliary neurotrophic factor (CNTF) , an agent which has previously been shown to induce the astrocytic differentiation of neural stem cells (NSC) in vitro.
- CNTF .ciliary neurotrophic factor
- the GBM cell line U343 MG-A was treated with 50 ⁇ M of retinoic acid (RA) and/or 1OnM of okadaic acid (OA) , a protein phosphatase-1 inhibitor.
- RA retinoic acid
- OA 1OnM of okadaic acid
- GBM glioblastoma multiforme
- N- CoR nuclear receptor co-repressor
- N-CoR nuclear expression of N-CoR correlated with the absence of GFAP expression in the cells of primary cultures and tissue sections, whereas cytoplasmic expression of N-CoR correlated with positive expression of GFAP.
- N-CoR neurotrophic factor
- BTSCs brain tumor stem cells
- CNTF brain tumor stem cells
- Western blot analysis of the cytoplasmic fraction of CNTF-treated BTSCs demonstrates the translocation of N-CoR to the cytoplasm (See Figure 2B) .
- Both cytoplasmic N-CoR and GFAP expression peaked at day 7 following CNTF stimulation.
- Figure 2C shows the logarithmic growth curve of gliomal cell line, U343 MG-A, treated with retinoic acid (RA) , okadaic acid (OA) , combination of retinoic acid and okadaic acid (RA/OA) , and control (NC) for 16 days.
- Curve fitting indicated exponential cell count growth for each treatment except for the RA/OA treatment group.
- GBM glioblastoma multiforme
- the cantharidin homologs that were evaluated were norcantharidin (nor-Can) , which is a demethylated cantharidin; endothal (End) , which is a dicarboxylic acid derivative of norcantharidin; endothal thioanhydride (ET) ; and the compound LB-I, which was obtained from Lixte Biotechnology, Inc., 248 Route 25A, No. 2, East Setauket, New York, which has the structure:
- Cells were plated in triplicate on day one with and without different amounts of each drug dissolved in media (compound LB-I and endothal) or in dimethylsulfoxide (endothal thioanhydride and norcantharidin) . The total number of cells is counted in the triplicate cultures at each dose and in controls after 7 days and the average number of cells and the standard deviation is determined.
- the amount of inhibition of GBM cell growth is expressed as the proportion of the number of cells in the experimental dishes compared to the number of cells in control dishes containing only the drug vehicle and culture medium.
- the average percent of control is plotted and bracketed by one standard deviation calculated from the triplicate measurements.
- endothal was combined with all-trans retinoic acid and 13-cis retinoic acid.
- Cells were plated in triplicate on day one with and without different amounts of each drug dissolved in media (compound LB-I and endothal) . The total number of cells is counted in the triplicate cultures at each dose and in controls after 7 days and the average number of cells and the standard deviation is ' determined. .
- the amount of inhibition of GBM cell growth is expressed as the proportion of the number of cells in the experimental dishes compared to the number of cells in control dishes containing only the drug vehicle and culture medium.
- the average percent of control is plotted and bracketed by one standard deviation calculated from the triplicate measurements .
- Figure 4A demonstrates the effect of all-trans retinoic acid (ATRA) when used individually.
- ATRA all-trans retinoic acid
- the IC50 of ATRA alone was greater than 50 ⁇ M.
- Endothal and compound LB-I, each in combination with ATRA synergistically inhibited proliferation of GBM cell line U373 as seen in figure 4B.
- Synergism (potentiation), of the inhibitory activity of two drugs in combination is said to be present when the percent survival in the presence of two drugs is less than the product of the percent survivals of the two drugs used alone at the same doses in the combination.
- the extent of synergism of compound LB-I and endothal (end) in combination with ATRA is quantified below in Table 1:
- retinoids are known to produce developmental abnormalities in the fetus when the drug is given to pregnant women, we studied the activity of valproic acid and Trichostatin A, drugs with low toxicity in the adult, but which also disrupt fetal development.
- valproic acid (VaI) ( Figure 5A) and Trichostantin A (TSA) ( Figure 5B) had dose dependent activity as a single agent against U373 cell growth. Although inhibitory doses of valproic acid were in the mM range, the antiepileptic drug is tolerated in humans at serum concentrations approaching 1.0 mM for weeks. Trichostatin A, in contrast, is active at nM concentrations against U373.
- the two drugs are synergistic in their inhibition of the growth of U373 cells.
- the percent survival of the cells after exposure to two drugs in combination is less than would be expected from the percent survival of the cells when exposed to each of the two drugs at the same doses used in the combination .
- Example 5 Determination of tumor type specificity.
- the kidney cancer cell line, UMRC (Figure 6A) was less sensitive than the brain tumor line, U373 ( Figure 6B) whereas the breast cancer line, MCF-7 ( Figure 6C) was as sensitive as U373 to all-trans retinoic acid, endothal thioanhydride, norcantharidin, endothal, and Trichostatin A.
- MCF-7 ( Figure 6C) was as sensitive as U373 to all-trans retinoic acid, endothal thioanhydride, norcantharidin, endothal, and Trichostatin A.
- proteomes of 7 GBM tissues and 7 normal brain tissues were compared using selective microdissection, two dimensional gel electrophoresis (2-DGE) and liquid chromatography-mass spectroscopy (LCMS) .
- N- CoR nuclear receptor co-repressor
- N-CoR is expressed in the nucleus of neural stem cells (NSCs) .
- NSCs neural stem cells
- BTSC brain tumor stem cells
- BTSC are capable of proliferation, self-renewal, and differentiation.
- BTSC but not CD133- differentiated tumor cells, are able to recapitulate tumors upon xenograft transplantation.
- N-CoR glial fibrillary acidic protein
- N-CoR Nuclear expression of N-CoR correlated with the absence of GFAP expression in the cells of primary cultures and tissue sections (see Figure ID) , whereas cytoplasmic expression of N-CoR correlated with positive expression of GFAP (see Figure ID) . Some of the tumor cells with nuclear expression of N-CoR also expressed CD133 (see Figure IE) .
- N-CoR ciliary neurotrophic factor
- N-CoR is closely associated with the retinoid receptor and is released upon ligand binding to the receptor.
- Bastien et al. (2004) We hypothesized that one effect of retinoids on malignant gliomas may be the induction of differentiation by the binding of retinoids to the retinoid receptor followed by dissociation of the N-CoR/retinoid receptor complex and translocation of N-CoR to the cytoplasm. This idea would explain the previous observation of increase GFAP expression in a glioma cell line (U343 MG-A) treated with retinoids.
- okadaic acid that have anti-PP2A activity synergize with all-trans retinoic acid and 13-cis retinoic acid in inhibiting the growth of GBM cells in vitro.
- the most effective group of phosphatase inhibitors synergizing with retinoic acids that have been evaluated are analogs of the ancient therapeutic agent, mylabris, derived from the crushed bodies of the blister beetle, in which the principal active agent is cantharidin, a known potent inhibitor of PP2A (Wang, 1989; Peng et al., 2002).
- Cantharidin has anti-tumor activity against human cancers of the liver (hepatomas) and of the upper gastrointestinal tract but is toxic to the urinary tract (Wang, 1989) .
- Norcantharidin a demethylated cantharidin, maintains antitumor activity of cantharidin against hepatomas and cancers of the stomach and esophagus, but has little or no urinary tract toxicity.
- Norcantharidin increased the life span of 244 patients with primary hepatoma from 4.7 to 11.1 months and increased 1-year survival from 17% to 30% compared to historical control patients treated with standard chemotherapy.
- Norcantharidin also stimulates white blood cell production in patients and mice, a phenomenon not understood mechanistically, but a pharmacological effect of potential benefit as an anticancer agent (Wang et al., 1986; Wang, 1989) .
- Endothal and endothal thioanhydride like cantharidin, inhibit the activity of PP2A and to some extent, the activity of PPl
- GBM glioblastoma multiforme
- compound LB-I norcantharidin (nor-Can)
- endothal End
- ET endothal thioanhydride
- IC50 concentration of each compound that inhibited brain tumor cell proliferation by 50%
- IC50s expressed in micro-molarity ( ⁇ M) were: 2.5, 3.0, 12.0, and 15.0 for endothal thioanhydride, compound LB-I, norcantharidin, and endothal respectively as seen in figure 3.
- endothal thioanhydride was the most potent inhibitor of GBMs in vitro compared to norcantharidin and endothal.
- retinoids synergistically inhibit the proliferation of glioblastoma multiforme.
- Synergism (potentiation) of the inhibitory activity of two drugs in combination is said to be present when the percent survival in the presence of two drugs is less than the product of the percent survivals of the two drugs used alone at the same doses in the combination.
- the inhibitory activity of retinoids was further evaluated in combination with endothal as well as individually.
- increase in the dose of ATRA exhibits inhibitory activity on glial cancer cells.
- the combination of endothal with ATRA demonstrated a synergistic reduction in cell growth.
- the expected percent survival in the presence of the combination of ATRA and LB-I is 60% (77% by ATRA x 78% by LB-I - 60%), whereas the observed survival was 53%.
- Trichostatin A is a natural product extracted from streptomyces, which has anti-fungal and anti-cancer activity in vitro and in human cancer xenografts (Yoshida et al . , 1990; Sanderson et al., 2004) .
- Valproic acid is a widely used anti- seizure medicine that inhibits human cancer cells in vitro at concentrations achievable in the plasma of humans (G ⁇ ttiere et al., 2001; . Blaheta et al., 2002).
- both valproic acid (VaI) and Trichostatin A (TSA) had dose dependent activity as single agents against U373 cell growth.
- Trichostatin A is active at nM concentrations against U373. Given the low toxicity in non-pregnant adults, both compounds combined with endothal could be potentially effective regimens in the treatment of GBM in humans .
- Cantharidin homologs and okadaic acid act synergistically when administered with valproic acid or trichostatin A to inhibit growth of GBM cells. Both valproic acid and trichostatin A are known to have anti-histone deacetylase (HDAC) activity.
- HDAC anti-histone deacetylase
- retinoic acid and Trichostatin A we measured their inhibitory effects as single agents against the GBM line U373, a breast cancer line, MCF-7 (obtained from ATCC) and a kidney cancer cell line, UMRC (UMRC obtained by Dr. Zhuang, NINDS, NIH from the Intramural Research Support Program, SAIC, National Cancer Institute, Frederick Cancer Research and Development Center) .
- kidney cancer cell line, UMRC (Figure 6A) was less sensitive than the brain tumor line, U373 ( Figure 6B) whereas the breast cancer line, MCF-7 ( Figure 6C) was as sensitive as
- N-CoR pathway targeting induces glioblastoma derived cancer stem cell differentiation, Cell Cycle, in press.
Abstract
Description
Claims
Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US77116306P | 2006-02-06 | 2006-02-06 | |
US79720106P | 2006-05-02 | 2006-05-02 | |
PCT/US2007/003095 WO2007092414A2 (en) | 2006-02-06 | 2007-02-06 | Use of phosphatases to treat tumors overexpressing n-cor |
Publications (2)
Publication Number | Publication Date |
---|---|
EP1986619A2 true EP1986619A2 (en) | 2008-11-05 |
EP1986619A4 EP1986619A4 (en) | 2010-04-28 |
Family
ID=38345726
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
EP07763614A Withdrawn EP1986619A4 (en) | 2006-02-06 | 2007-02-06 | Use of phosphatases to treat tumors overexpressing n-cor |
Country Status (3)
Country | Link |
---|---|
EP (1) | EP1986619A4 (en) |
CA (1) | CA2641308A1 (en) |
WO (1) | WO2007092414A2 (en) |
Families Citing this family (11)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2008097561A1 (en) * | 2007-02-06 | 2008-08-14 | Lixte Biotechology Holdings, Inc. | Oxabicycloheptanes and oxabicycloheptenes, their preparation and use |
EP2185173A4 (en) * | 2007-08-03 | 2011-01-12 | Lixte Biotechnology Inc | Use of phosphatases to treat neuroblastomas and medullogastomas |
EP2200439B1 (en) | 2007-10-01 | 2017-03-22 | Lixte Biotechnology, Inc. | Hdac inhibitors |
CA2730428A1 (en) * | 2008-08-01 | 2010-02-04 | Lixte Biotechnology, Inc. | Methods for regulating cell mitosis by inhibiting serine/threonine phosphatase |
WO2010147612A1 (en) * | 2009-06-18 | 2010-12-23 | Lixte Biotechnology, Inc. | Methods of modulating cell regulation by inhibiting p53 |
CA2827015A1 (en) * | 2010-02-11 | 2011-08-18 | The Royal Institution For The Advancement Of Learning/Mcgill University | Hybrid molecule having mixed retinoic acid receptor agonism and histone deacetylase inhibitory properties |
CA2877167A1 (en) | 2012-06-29 | 2014-01-03 | Lixte Biotechnology, Inc. | Oxabicycloheptanes and oxabicycloheptenes for the treatment of diabetes |
EA201591931A1 (en) | 2013-04-09 | 2016-05-31 | Ликсте Байотекнолоджи, Инк. | COMPOSITIONS OF OXABICYCLOPTANES AND OXABITICLOCEPTENS |
JP6453441B2 (en) | 2014-07-24 | 2019-01-16 | エイチ リー モフィット キャンサー センター アンド リサーチ インスティテュート インコーポレイテッド | Protein phosphatase 2A inhibitor for the treatment of myelodysplastic syndrome |
WO2016134257A1 (en) | 2015-02-19 | 2016-08-25 | Lixter Biotechnology, Inc. | Oxabicycloheptanes and oxabicycloheptenes for the treatment of depressive and stress disorders |
EP3294287B1 (en) | 2015-05-15 | 2020-04-08 | Lixte Biotechnology, Inc. | Oxabicycloheptane prodrugs |
Citations (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US6262116B1 (en) * | 1998-01-23 | 2001-07-17 | Sloan-Kettering Institute For Cancer Research | Transcription therapy for cancers |
WO2002009680A2 (en) * | 2000-08-02 | 2002-02-07 | Michael Walter | Pharmacologically active substance for treating cardiovascular disorders |
WO2002028387A1 (en) * | 2000-10-03 | 2002-04-11 | Oncopharmaceutical, Inc. | Inhibitors of angiogenesis and tumor growth for local and systemic administration |
WO2003092719A2 (en) * | 2002-04-29 | 2003-11-13 | Yissum Research Development Company Of The Hebrew University Of Jerusalem | Methods and compositions for modulating beta-catenin phosphorylation |
US20040209934A1 (en) * | 2001-03-23 | 2004-10-21 | Mccluskey Adam | Protein phosphate inhibitors |
FR2872704A1 (en) * | 2004-07-12 | 2006-01-13 | Laurent Schwartz | PLURITHERAPY AGAINST CANCER |
-
2007
- 2007-02-06 WO PCT/US2007/003095 patent/WO2007092414A2/en active Application Filing
- 2007-02-06 EP EP07763614A patent/EP1986619A4/en not_active Withdrawn
- 2007-02-06 CA CA002641308A patent/CA2641308A1/en not_active Abandoned
Patent Citations (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US6262116B1 (en) * | 1998-01-23 | 2001-07-17 | Sloan-Kettering Institute For Cancer Research | Transcription therapy for cancers |
WO2002009680A2 (en) * | 2000-08-02 | 2002-02-07 | Michael Walter | Pharmacologically active substance for treating cardiovascular disorders |
WO2002028387A1 (en) * | 2000-10-03 | 2002-04-11 | Oncopharmaceutical, Inc. | Inhibitors of angiogenesis and tumor growth for local and systemic administration |
US20040209934A1 (en) * | 2001-03-23 | 2004-10-21 | Mccluskey Adam | Protein phosphate inhibitors |
WO2003092719A2 (en) * | 2002-04-29 | 2003-11-13 | Yissum Research Development Company Of The Hebrew University Of Jerusalem | Methods and compositions for modulating beta-catenin phosphorylation |
FR2872704A1 (en) * | 2004-07-12 | 2006-01-13 | Laurent Schwartz | PLURITHERAPY AGAINST CANCER |
Non-Patent Citations (5)
Title |
---|
FRASOR JONNA ET AL: "Estrogen down-regulation of the corepressor N-CoR: Mechanism and implications for estrogen derepression of N-CoR-regulated genes" PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, vol. 102, no. 37, September 2005 (2005-09) , pages 13153-13157, XP002572866 ISSN: 0027-8424 * |
HILL ET AL: "Heterocyclic substituted cantharidin and norcantharidin analogues-synthesis, protein phosphatase (1 and 2A) inhibition, and anti-cancer activity" BIOORGANIC & MEDICINAL CHEMISTRY LETTERS, PERGAMON, ELSEVIER SCIENCE, GB, vol. 17, no. 12, 15 June 2007 (2007-06-15) , pages 3392-3397, XP022097790 ISSN: 0960-894X * |
KOK S-H ET AL: "Norcantharidin-induced apoptosis in oral cancer cells is associated with an increase of proapoptotic to antiapoptotic protein ratio" CANCER LETTERS, NEW YORK, NY, US, vol. 217, no. 1, 10 January 2005 (2005-01-10), pages 43-52, XP004681287 ISSN: 0304-3835 * |
LEE MI-OCK ET AL: "Role of coactivators and corepressors in the induction of the RARbeta gene in human colon cancer cells." BIOLOGICAL AND PHARMACEUTICAL BULLETIN, vol. 25, no. 10, October 2002 (2002-10), pages 1298-1302, XP002572865 ISSN: 0918-6158 * |
See also references of WO2007092414A2 * |
Also Published As
Publication number | Publication date |
---|---|
EP1986619A4 (en) | 2010-04-28 |
WO2007092414A8 (en) | 2008-09-04 |
WO2007092414A3 (en) | 2008-05-15 |
WO2007092414A2 (en) | 2007-08-16 |
CA2641308A1 (en) | 2007-08-16 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
US20080214569A1 (en) | Use of phosphatases to treat tumors overexpressing N-CoR | |
EP1986619A2 (en) | Use of phosphatases to treat tumors overexpressing n-cor | |
Burnstock et al. | Purinergic signaling in healthy and diseased skin | |
JP5620443B2 (en) | Vitamin K for prevention and treatment of rash secondary to anti-EGFR therapy | |
KR102473113B1 (en) | Combination therapy for treating cancer | |
Huang et al. | Capsaicin protects cortical neurons against ischemia/reperfusion injury via down-regulating NMDA receptors | |
US20090035292A1 (en) | Use of phosphatases to treat neuroblastomas and medulloblastomas | |
JP2017507151A (en) | Use of eribulin and mTOR inhibitors as combination therapy for the treatment of cancer | |
EP3258930B1 (en) | Oxabicycloheptanes and oxabicycloheptenes for the treatment of depressive and stress disorders | |
TW202122084A (en) | Methods and materials for treating neurotoxicity | |
Kyriakou et al. | Efficacy of cannabinoids against glioblastoma multiforme: A systematic review | |
Da Fonseca et al. | Anaplastic oligodendroglioma responding favorably to intranasal delivery of perillyl alcohol: a case report and literature review | |
EP3989963A1 (en) | Carbocyanine compounds for targeting mitochondria and eradicating cancer stem cells | |
US20130177627A1 (en) | Growth inhibitory effects of nanoparticles containing triterpene glycosides or triterpenes | |
IL308668A (en) | Composition for treating autoimmune, alloimmune, inflammatory, and mitochondrial conditions, and uses thereof | |
US20170136053A1 (en) | Novel pharmaceutical composition and uses thereof | |
JP7296119B2 (en) | Combination therapy of safranal and sorafenib for liver cancer | |
US20180243299A1 (en) | Compositions and methods for treating ewing family tumors | |
AU2021207766A1 (en) | Cannabinoids for use in treatment | |
US11382892B2 (en) | Method for administration | |
US20150209364A1 (en) | Method for treating epidermoid carcinoma | |
KR20090047501A (en) | Methods and pharmaceutical preparations for contributing to the treatment of chemotherapy-induced neuropathy | |
WO2011066545A1 (en) | Treatment of human osteosarcoma |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
PUAI | Public reference made under article 153(3) epc to a published international application that has entered the european phase |
Free format text: ORIGINAL CODE: 0009012 |
|
17P | Request for examination filed |
Effective date: 20080904 |
|
AK | Designated contracting states |
Kind code of ref document: A2 Designated state(s): AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HU IE IS IT LI LT LU LV MC NL PL PT RO SE SI SK TR |
|
AX | Request for extension of the european patent |
Extension state: AL BA HR MK RS |
|
RIN1 | Information on inventor provided before grant (corrected) |
Inventor name: KOVACH, JOHN, S. Inventor name: LI, JIE Inventor name: LUBENSKY, IRINA Inventor name: PARK, DERIC, M. Inventor name: OLDFIELD, EDWARD, H. Inventor name: ZHUANG, ZHENGPING |
|
RAP1 | Party data changed (applicant data changed or rights of an application transferred) |
Owner name: THE GOVERNMENT OF THE UNITED STATES OF AMERICA A Owner name: LIXTE BIOTECHNOLOGY, INC. |
|
RIC1 | Information provided on ipc code assigned before grant |
Ipc: A61P 35/00 20060101ALI20100318BHEP Ipc: A61K 31/00 20060101AFI20080912BHEP |
|
A4 | Supplementary search report drawn up and despatched |
Effective date: 20100330 |
|
17Q | First examination report despatched |
Effective date: 20110411 |
|
DAX | Request for extension of the european patent (deleted) | ||
GRAP | Despatch of communication of intention to grant a patent |
Free format text: ORIGINAL CODE: EPIDOSNIGR1 |
|
RIC1 | Information provided on ipc code assigned before grant |
Ipc: A61K 31/519 20060101ALI20140610BHEP Ipc: A61P 35/00 20060101ALI20140610BHEP Ipc: A61K 31/00 20060101AFI20140610BHEP |
|
INTG | Intention to grant announced |
Effective date: 20140703 |
|
STAA | Information on the status of an ep patent application or granted ep patent |
Free format text: STATUS: THE APPLICATION IS DEEMED TO BE WITHDRAWN |
|
18D | Application deemed to be withdrawn |
Effective date: 20141114 |