WO2005053704A1 - Procede de prediction de sensibilite aux medicaments dans des neoplasmes myeloides - Google Patents

Procede de prediction de sensibilite aux medicaments dans des neoplasmes myeloides Download PDF

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WO2005053704A1
WO2005053704A1 PCT/EP2004/013588 EP2004013588W WO2005053704A1 WO 2005053704 A1 WO2005053704 A1 WO 2005053704A1 EP 2004013588 W EP2004013588 W EP 2004013588W WO 2005053704 A1 WO2005053704 A1 WO 2005053704A1
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apoptosis
patient
treatment
compounds
cells
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PCT/EP2004/013588
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Kenneth Wayne Culver
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Novartis Ag
Novartis Pharma Gmbh
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    • 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
    • A61K31/55—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having seven-membered rings, e.g. azelastine, pentylenetetrazole
    • A61K31/553—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having seven-membered rings, e.g. azelastine, pentylenetetrazole having at least one nitrogen and one oxygen as ring hetero atoms, e.g. loxapine, staurosporine
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A61P35/02—Antineoplastic agents specific for leukemia

Definitions

  • This invention relates generally to the analytical testing of tissue samples in vitro, and more particularly to methods for predicting the responsiveness of patients with myeloid neoplasms to chemotherapy agents.
  • this invention relates to the use of apoptosis assays performed on peripheral blood and bone marrow samples to predict patient response to any cancer therapeutic agent that induces apoptosis as part of it's mechanism of action, including tyrosine kinase inhibitors, such as staurosporine derivatives including N-benzoylstaurosporine (midostaurin).
  • the myeloid neoplasms are primary malignant diseases of the blood-forming organs and are characterized by a predominance of immature myeloid precursors (blasts). The blasts progressively replace normal bone marrow, migrate and invade other tissues. There is diminished production of normal erythrocytes, granulocytes and platelets in patients with acute leukaemia, and this leads to the most important complications of this disease, i.e., anaemia, infection and haemorrhage.
  • AML acute myeloid leukaemia
  • Acute leukaemia is the result of a malignant event or events occurring in an early haematopoietic precursor. Instead of proliferating and differentiating normally, the affected cell gives rise to progeny that fail to differentiate and instead continue to proliferate in an uncontrolled fashion. As a result, immature myeloid cells in AML or lymphoid cells in ALL, (blasts), rapidly accumulate and progressively replace the bone marrow; causing diminished production of normal red cells, white cells and platelets.
  • This may include radiation; oncogenic viruses including retroviruses, such as HTLV-1 ; genetic and congenital factors; chemicals, such as benzene and kerosene; and drugs, such as epipodophyllotoxins.
  • oncogenic viruses including retroviruses, such as HTLV-1 ; genetic and congenital factors; chemicals, such as benzene and kerosene; and drugs, such as epipodophyllotoxins.
  • drugs such as epipodophyllotoxins.
  • Tyrosine Kinase Inhibitors including but not limited to, imatinib mesylate and staurosporine derivatives such as N-benzoylstaurosporine (midostaurin; PKC412); HDAC inhibitors; B-Raf inhibitors, including the biaryl isoquinoline group; FAK inhibitors; Edg-1 inhibitors; IAP inhibitors; Hepararanase inhibitors; Protein Tyrosine Phosphatese inhibitors; CXCR4 inhibitors; DNA methyl transferase inhibitors; PPARgamma and alpha agonists; epothilones; Edg-4 receptor antagonists and many others, see below.
  • TKIs Tyrosine Kinase Inhibitors
  • imatinib mesylate and staurosporine derivatives such as N-benzoylstaurosporine (midostaurin; PKC412)
  • HDAC inhibitors B-Raf inhibitors, including the biaryl isoquinoline group
  • This invention provides a solution for the above mentioned problem.
  • this invention provides a method to estimate the response of a patient with a myeloid neoplasm to treatment with an apoptosis inducing chemotherapy agent comprising: (a) measuring the extent of apoptosis in the blasts in a patient sample; and (b) determining from the measurement in Step (a) the likelihood that the patient will respond to treatment with an apoptosis inducing chemotherapy agent.
  • this invention provides a method wherein the patient sample consists of a smear of blood or a smear of bone marrow aspirate or cytospin of aspirate.
  • this invention provides a method wherein the apoptosis inducing chemotherapy agent is a TKI including, but not limited to, N-benzoylstaurosporine.
  • the method of the invention involves the determination step above consists of finding that the patient is a responder if the percentage of apoptosis is greater than 5%.
  • this invention provides a method to determine if a patient with a myeloid neoplasm should be included in a drug study of an apoptosis inducing chemotherapy agent comprising: (a) measuring the extent of apoptosis in the blasts in a patient sample; and (b) determining from the measurement in Step (a) if the patient will respond to treatment with an apoptosis inducing chemotherapy agent and should be included in the study.
  • This method may make use of a patient sample consisting of a smear of blood or of a smear of bone marrow aspirate or cytospin of aspirate. In this method the patient will be considered to be a responder if the percentage of apoptosis is greater than 5%.
  • Another aspect of the invention relates to the use of N-benzoylstaurosporine in the manufacture of a medicament for the treatment of myeloid neoplasms in a selected patient population, wherein the patient population is selected on the basis of a measurement of the pre-treatment extent of apoptosis in the blasts in a patient sample.
  • the present invention provides methods to estimate or predict which patients with a myeloid neoplasm will show a favourable response to an apoptosis inducing chemotherapy agent. These methods, to predict drug resistance and drug response can be used to guide the choice of therapy for a particular patient, help determine drug dose and schedule and can be used to determine the correct patients to include in a drug study.
  • This invention is based, in part, on the discovery that the extent of apoptosis seen in blast cells in peripheral blood or bone marrow of patients with myeloid neoplasm, including
  • AML prior to treatment with an apoptosis inducing therapeutic agent can predict the patients response to that agent.
  • the leukaemias account for about 3% of all cancers in the U.S.
  • the impact of leukaemia is heightened because of the young age of some patients.
  • ALL is the most common cancer and the second leading cause of death in children younger than 15 years.
  • ALL has a maximal incidence between 2 and 10 years of age, with a second, more gradual rise in frequency later in life.
  • the incidence of AML gradually increases with age, without an early peak. Approximately half of AML cases occur in patients younger than 50 years.
  • malignancy occurs in a very undifferentiated cell, similar to the normal haematopoietic stem cell, in this case, cells, platelets and myeloid precursors are all products of the malignant clone.
  • the malignant event may occur in a more differentiated cell; therefore granulocyte and monocyte precursors will develop from the malignant cell, but red cells and platelet precursors will not.
  • the myeloid lineage is not malignant, which suggests that in ALL the malignant event occurs in a cell that is at least partially differentiated.
  • some mature circulating cells may be products of the malignant clone.
  • the acute leukaemias can be classified in a variety of ways, including morphology, cytochemistry, cell-surface markers, cytoplasmic markers, cytogenetics and oncogene expression. The most important distinction is between AML and ALL because these two diseases differ considerably in their clinical behaviour, prognosis, and response to therapy. The various subgroups of AML and ALL also have some important differences. [22] Morphology. Leukaemic cells in AML are typically 12-20 mm in diameter, with discrete nuclear chromatin, multiple nucleoli and cytoplasm that usually contains azurophilic granules.
  • Auer rods which are slender, fusiform cytoplasmic inclusions that stain red with Wright-Giemsa stain, are virtually pathognomonic of AML.
  • the French-American-British (FAB) collaborative group has subdivided AML into eight subtypes based on morphology and histochemistry: • M0, M1 , M2 and M3 - reflect increasing degrees of differentiation of myeloid leukaemic cells; • M4 and M5 - features of the monocytic lineage; • M6 - features of the erythroid cell lineage; and • M7 - acute megakaryocytic leukaemia.
  • lymphoid antigens by AML cells does not change either the natural history or the therapeutic response of these leukaemias.
  • Cytogenetics and molecular biology In most cases of acute leukaemia, an abnormality in chromosome number or structure is found. These abnormalities are clonal, essentially involving all of the malignant cells in a given patient, are acquired and not found in the normal cells of the patient, and are referred to as "non-random" because specific abnormalities are found in multiple cases of AML and are associated with distinct morphologic or clinical subtypes of the disease. These abnormalities may be simply the gain or loss of whole chromosomes, but more often they include chromosomal translocations, deletions or inversions. When patients with acute leukaemia and a chromosomal abnormality are treated and enter into complete remission, the chromosomal abnormality disappears; when relapse occurs, the abnormality reappears.
  • Apoptosis is programmed cell death. During development and later, excess numbers of neurons, lymphocytes and many other kinds of cells die through a genetically programmed sequence of changes called apoptosis. This phenomena is of vital importance in determining the malignancy and resistance to treatment of neoplastic cells.
  • the term apoptosis was coined by in 1972 as a means of distinguishing a morphologically distinctive form of cell death which was associated with normal physiology. See Kerr, Wyllie & Currie, BrJ Cancer 26(24): 239-257 (1972). Apoptosis was distinguished from necrosis, which was associated with acute injury to cells.
  • Apoptosis is characterized by nuclear chromatin condensation, cytoplasmic shrinking, dilated endoplasmic reticulum and membrane blebbing, mitochondria remain unchanged morphologically.
  • This type of cell death is often hard to observe in vivo because the dying cells are rapidly phagocytosed by tissue macrophages, and this phagocytosis is clearly different from that seen in inflammation, when activated macrophages are recruited from outside the immediate area of death.
  • One way to observe this phenomenon in vitro is to use a cell permeant DNA-staining fluorescent dye, such as Hoechst 33342, which allows a striking visualization of the chromatin condensation.
  • Apoptotic death can be triggered by a wide variety of stimuli, and not all cells necessarily will die in response to the same stimulus.
  • DNA damage by irradiation or drugs used for cancer chemotherapy
  • Some hormones, such as corticosteroids lead to death in particular cells, e.g., thymocytes, although other cell types may be stimulated.
  • Some cells types express Fas, a surface protein which initiates an intracellular death signal in response to cross-linking. In other cases cells appear to have a default death pathway which must be actively blocked by a survival factor in order to allow cell survival. When the survival factor is removed, the default apoptotic death program is triggered.
  • apoptotic systems e.g., Fas killing of tumour cells
  • artificially enucleated cells lacking a nucleus still die, showing that the nucleus is not always necessary for apoptotic cell death.
  • the changes in the apoptotic cell trigger phagocytosis by non-activated macrophages. Macrophages appear to recognize apoptotic cells via several different recognition systems. For example, there is good evidence that apoptotic cells lose the normal phospholipid asymmetry in their plasma membrane, as manifested by the exposure of normally inward-facing phosphatidyl serine on the external face of the bilayer.
  • Macrophages can recognize this exposed lipid headgroup via an unknown receptor, triggering phagocytosis. Exposure of phosphatidyl serine on the surface of apoptotic cells is depicted in the right blow-up at the bottom of the diagram above.
  • caspases are cysteine proteases related to ced-3, the "death gene" of the nematode Caenorhabditis elegans.
  • Caspases seem to be widely-expressed in an inactive proenzyme form in most cells.
  • Their proteolytic activity is characterized by their unusual ability to cleave proteins at aspartic acid residues, although different caspases have different fine specificities involving recognition of neighbouring amino acids. Active caspases can often activate other pro-caspases, allowing initiation of a protease cascade.
  • apoptosis inducing chemotherapy agent shall mean any compound, means or agent for use in treating a neoplastic disease that directly or indirectly induces apoptosis as part of the anti-tumour effect. This is intended to include all types of pharmacological therapeutics or other means or agents for treating a neoplastic disease except for immunotherapies and vaccines.
  • This group of therapeutic agents is a broad one covering many chemotherapeutic agents having different mechanisms of action. Generally, chemotherapeutic agents are classified according to the mechanism of action. Many of the available agents are anti-metabolites of development pathways of various tumours, or react with the DNA of the tumour cells. There are also agents which inhibit enzymes, such as topoisomerase I and topoisomerase II, or which are antimiotic agents.
  • apoptosis inducing chemotherapy agent is meant, in addition, any chemotherapeutic agent including, but not limited to: i. an aromatase inhibitor; ii. an antioestrogen, an anti-androgen (especially in the case of prostate cancer) or a gonadorelin agonist; iii. a topoisomerase I inhibitor or a topoisomerase II inhibitor; iv. a microtubule active agent, an alkylating agent, an anti-neoplastic anti- metabolite or a platin compound; v.
  • any chemotherapeutic agent including, but not limited to: i. an aromatase inhibitor; ii. an antioestrogen, an anti-androgen (especially in the case of prostate cancer) or a gonadorelin agonist; iii. a topoisomerase I inhibitor or a topoisomerase II inhibitor; iv. a microtubule active agent, an alkylating
  • a compound targeting/decreasing a protein or lipid kinase activity or a protein or lipid phosphatase activity, a further anti-angiogenic compound or a compound which induces cell differentiation processes vi. a bradykinin 1 receptor or an angiotensin II antagonist; vii. a cyclooxygenase inhibitor, a bisphosphonate, a rapamycin derivative, such as everolimus, a heparanase inhibitor (prevents heparan sulphate degradation), e.g., PI-88, a biological response modifier, preferably a lymphokine or interferons, e.g.
  • interferon ⁇ an ubiquitination inhibitor or an inhibitor which blocks anti-apoptotic pathways
  • viii an inhibitor of Ras oncogenic isoforms or a famesyl transferase inhibitor
  • ix a telomerase inhibitor, e.g., telomestatin
  • x. a protease inhibitor, a matrix metalloproteinase inhibitor, a methionine aminopeptidase inhibitor, e.g., bengamide or a derivative thereof; or a proteosome inhibitor, e.g., PS-341
  • agents used in the treatment of haematologic malignancies or FMS-like tyrosine kinase inhibitors xii.
  • HSP90 inhibitors an HSP90 inhibitors
  • xiii histone deacetylase (HDAC) inhibitors
  • xiv serine/theorine mTOR inhibitors
  • xv somatostatin receptor antagonists
  • xvi. ocv ⁇ 3/5 integrin antagonists xvii. anti-leukaemic compounds
  • tumour cell damaging approaches such as ionizing radiation
  • xix. EDG binders xx. antranilamide class of compounds
  • xxii S-adenosylmethionine decarboxylase inhibitors
  • VEGF monoclonal antibodies of VEGF or VEGFR; or a VEGF inhibitor or a derivative thereof.
  • VEGF vascular endothelial growth factor
  • aromatase inhibitor relates to a compound which inhibits the oestrogen production, i.e., the conversion of the substrates androstenedione and testosterone to oestrone and oestradiol, respectively.
  • the term includes, but is not limited to, steroids, especially atamestane, exemestane and formestane; and, in particular, non- steroids, especially aminoglutethimide, roglethimide, pyridoglutethimide, trilostane, testolactone, ketokonazole, vorozole, fadrozole, anastrozole and letrozole.
  • Exemestane is marketed as AROMASIN ® ; formestane as LENTARON ® ; fadrozole as AFEMA ® ; anastrozole as ARIMIDEX ® ; letrozole as FEMARA ® or FEMAR ® ; and aminoglutethimide as ORIMETEN ® .
  • a combination of the invention comprising a chemotherapeutic agent which is an aromatase inhibitor is particularly useful for the treatment of hormone receptor positive tumours, e.g., breast tumours.
  • anti-oestrogen relates to a compound which antagonizes the effect of estrogens at the oestrogen receptor level.
  • the term includes, but is not limited to, tamoxifen, fulvestrant, raloxifene and raloxifene hydrochloride.
  • Tamoxifen can be administered in the form as it is marketed, e.g., NOLVADEX ® ; and raloxifene hydrochloride is marketed as EVISTA ® .
  • Fulvestrant can be formulated as disclosed in U.S. Pat. No. 4,659,516 and is marketed as FASLODEX ® .
  • a combination of the invention comprising a chemotherapeutic agent which is an anti-oestrogen is particularly useful for the treatment of oestrogen receptor positive tumours, e.g., breast tumours.
  • anti-androgen as used herein, relates to any substance which is capable of inhibiting the biological effects of androgenic hormones and includes, but is not limited to, bicalutamide (CASODEX ® ), which can be formulated, e.g., as disclosed in U.S. Pat. No. 4,636,505.
  • gonadorelin agonist includes, but is not limited to, abarelix, goserelin and goserelin acetate. Goserelin is disclosed in U.S. Pat. No. 4,100,274 and is marketed as ZOLADEX ® . Abarelix can be formulated, e.g., as disclosed in U.S. Patent No. 5,843,901.
  • topoisomerase I inhibitor includes, but is not limited to, topotecan, irinotecan, camptothecian and its analogues, 9-nitrocamptothecin and the macromolecular camptothecin conjugate PNU-166148 (compound A1 in WO 99/17804).
  • Irinotecan can be administered, e.g., in the form as it is marketed, e.g., under the trademark CAMPTOSAR ® .
  • Topotecan can be administered, e.g., in the form as it is marketed, e.g., under the trademark HYCAMTIN ® .
  • topoisomerase II inhibitor includes, but is not limited to, the anthracyclines, such as doxorubicin, including liposomal formulation, e.g., CAELYX ® , daunorubicin, epirubicin, idarubicin and nemorubicin; the anthraquinones mitoxantrone and losoxantrone; and the podophillotoxines etoposide and teniposide.
  • Etoposide is marketed as ETOPOPHOS ® ; teniposide as VM 26-BRISTOL ® ; doxorubicin as ADRIBLASTIN ® or ADRIAMYCIN ® ; epirubicin as FARMORUBICIN ® ; idarubicin as ZAVEDOS ® ; and mitoxantrone as NOVANTRON ® .
  • microtubule active agent relates to microtubule stabilizing, microtubule destabilizing agents and microtublin polymerization inhibitors including, but not limited to, taxanes, e.g., paclitaxel and docetaxel; vinca alkaloids, e.g., vinblastine, especially vinblastine sulfate; vincristine, especially vincristine sulfate and vinorelbine; discodermolides; cochicine and epothilones; and derivatives thereof, e.g., epothilone B or a derivative thereof.
  • taxanes e.g., paclitaxel and docetaxel
  • vinca alkaloids e.g., vinblastine, especially vinblastine sulfate
  • vincristine especially vincristine sulfate and vinorelbine
  • discodermolides cochicine and epothilones
  • derivatives thereof e.g., epothilone
  • Paclitaxel may be administered, e.g., TAXOL ® ; docetaxel as TAXOTERE ® ; vinblastine sulfate as VINBLASTIN R.P ® ; and vincristine sulfate as FARMISTIN ® .
  • Discodermolide can be obtained, e.g., as disclosed in U.S. Pat. No. 5,010,099.
  • Epotholine derivatives which are disclosed in U.S. Pat. No. 6,194,181 , WO 98/10121 , WO 98/25929, WO 98/08849, WO 99/43653, WO 98/22461 and WO 00/31247.
  • Epotholine A and/or B are particularly preferred.
  • alkylating agent includes, but is not limited to, cyclophosphamide, ifosfamide, melphalan or nitrosourea (BCNU or Gliadel ® ).
  • Cyclophosphamide can be administered, e.g., in the form as it is marketed, e.g., under the trademark CYCLOSTIN ® ; and ifosfamide as HOLOXAN ® .
  • anti-neoplastic anti-metabolite includes, but is not limited to, 5- fluorouracil (5-FU); capecitabine; gemcitabine; DNA de-methylating agents, such as 5- azacytidine and decitabine; methotrexate; and edatrexate.
  • Capecitabine can be administered, e.g., in the form as it is marketed, e.g., under the trademark XELODA ® ; and gemcitabine as GEMZAR ® .
  • monoclonal antibody trastuzumab which can be administered, e.g., in the form as it is marketed, e.g., HERCEPTIN ® .
  • platinum compound includes, but is not limited to, carboplatin, cis-platin, cisplatinum and oxaliplatin.
  • Carboplatin can be administered, e.g., in the form as it is marketed, e.g., CARBOPLAT ® ; and oxaliplatin as ELOXATIN ® .
  • N- benzoylstaurosporine (midostaurin); examples of further compounds include, e.g., UCN-01 ; safingol; BAY 43-9006; Bryostatin 1 ; Perifosine; llmofosine; RO 318220 and RO 320432; GO 6976; Isis 3521 ; LY333531/LY379196; isochinoline compounds, such as those disclosed in WO 00/09495; FTIs; PD184352 or QAN697, a P13K inhibitor; xi) compounds targeting, decreasing or inhibiting the activity of protein-tyrosine kinase inhibitors, such as compounds which target, decrease or inhibit the activity of protein-tyrosine kinase inhibitors include imatinib mesylate (GLEEVEC ® ) or tyrphostin.
  • GLEEVEC ® imatinib mesylate
  • tyrphostin
  • a tyrphostin is preferably a low molecular weight (M r ⁇ 1500) compound, or a pharmaceutically acceptable salt thereof, especially a compound selected from the benzylidenemalonitrile class or the S-arylbenzenemalonirile or bisubstrate quinoline class of compounds, more especially any compound selected from the group consisting of Tyrphostin A23/RG-50810, AG 99, Tyrphostin AG 213, Tyrphostin AG 1748, Tyrphostin AG 490, Tyrphostin B44, Tyrphostin B44 (+) enantiomer, Tyrphostin AG 555, AG 494, Tyrphostin AG 556 and AG957 and adaphostin (4- ⁇ [(2,5- dihydroxyphenyl)methyl]amino ⁇ -benzoic acid adamantyl ester, NSC 680410, adaphostin); and xii) compounds targeting, decreasing or inhibiting the activity of the epidermal growth factor family
  • antibody is meant intact monoclonal antibodies, polyclonal antibodies, multi- specific antibodies formed from at least 2 intact antibodies, and antibodies fragments so long as
  • Compounds which target, decrease or inhibit the activity of a protein or lipid phosphatase are, e.g., inhibitors of phosphatase 1 , phosphatase 2A, PTEN or CDC25, e.g., okadaic acid or a derivative thereof.
  • Further anti-angiogenic compounds include compounds having another mechanism for their activity, e.g., unrelated to protein or lipid kinase inhibition, e.g., thalidomide (THALOMID) and TNP-470.
  • TAALOMID thalidomide
  • TNP-470 TNP-470.
  • Compounds which induce cell differentiation processes are e.g. retinoic acid, ⁇ -, ⁇ - or ⁇ -tocopherol or ⁇ - ⁇ - or ⁇ -tocotrienol.
  • cyclooxygenase inhibitor as used herein includes, but is not limited to, e.g., Cox-2 inhibitors, 5-alkyl substituted 2-arylaminophenylacetic acid and derivatives, such as celecoxib (CELEBREX ® ), rofecoxib (VIOXX ® ), etoricoxib, valdecoxib or a 5-alkyl-2- arylaminophenylacetic acid, e.g., 5-methyl-2-(2'-chloro-6'-fluoroanilino)phenyl acetic acid or lumiracoxib.
  • Cox-2 inhibitors such as celecoxib (CELEBREX ® ), rofecoxib (VIOXX ® ), etoricoxib, valdecoxib or a 5-alkyl-2- arylaminophenylacetic acid, e.g., 5-methyl-2-(2'-chloro-6'-fluoroanil
  • bisphosphonates includes, but is not limited to, etridonic, clodronic, tiludronic, pamidronic, alendronic, ibandronic, risedronic and zoledronic acid.
  • Etridonic acid can be administered, e.g., in the form as it is marketed, e.g.,
  • DIDRONEL ® clodronic acid as BONEFOS ® ; tiludronic acid as SKELID ® ; pamidronic acid as
  • heparanase inhibitor refers to compounds which target, decrease or inhibit heparin sulphate degradation.
  • the term includes, but is not limited to,
  • biological response modifier refers to a lymphokine or interferons, e.g., interferon ⁇ .
  • inhibitor of Ras oncogenic isoforms e.g., H-Ras, K-Ras or N-Ras, as used herein, refers to compounds which target, decrease or inhibit the oncogenic activity of
  • Ras e.g., a farnesyl transferase inhibitor (FTI), e.g., L-744832 or DK8G557.
  • FTI farnesyl transferase inhibitor
  • telomerase inhibitor refers to compounds which target, decrease or inhibit the activity of telomerase.
  • Compounds which target, decrease or inhibit the activity of telomerase are especially compounds which inhibit the telomerase receptor, e.g., telomestatin.
  • methionine aminopeptidase inhibitor refers to compounds which target, decrease or inhibit the activity of methionine aminopeptidase.
  • Compounds which target, decrease or inhibit the activity of methionine aminopeptidase are, e.g., bengamide or a derivative thereof.
  • proteosome inhibitor refers to compounds which target, decrease or inhibit the activity of the proteosome.
  • Compounds which target, decrease or inhibit the activity of the proteosome include, e.g., PS-341 and MLN 341.
  • matrix metalloproteinase inhibitor includes, but is not limited to, collagen peptidomimetic and non-peptidomimetic inhibitors; tetracycline derivatives, e.g., hydroxamate peptidomimetic inhibitor batimastat; and its orally-bioavailable analogue marimastat (BB-2516), prinomastat (AG3340), metastat (NSC 683551)
  • BMS-279251 BAY 12-9566, TAA211 , MMI270B or AAJ996.
  • agents used in the treatment of haematologic malignancies includes, but is not limited to, FMS-like tyrosine kinase inhibitors, e.g., compounds targeting, decreasing or inhibiting the activity of FMS-like tyrosine kinase receptors (Flt-3R); interferon, 1-b-D-arabinofuransylcytosine (ara-c) and bisulfan; and ALK inhibitors, e.g., compounds which target, decrease or inhibit anaplastic lymphoma kinase.
  • FMS-like tyrosine kinase inhibitors e.g., compounds targeting, decreasing or inhibiting the activity of FMS-like tyrosine kinase receptors (Flt-3R); interferon, 1-b-D-arabinofuransylcytosine (ara-c) and bisulfan
  • ALK inhibitors e.g., compounds which target, decrease or inhibit anaplastic lymphoma kinase
  • FMS-like tyrosine kinase receptors are especially compounds, proteins or antibodies which inhibit members of the Flt-3R receptor kinase family, e.g., N -benzoylstaurosporine (PKC412, midostaurin, a staurosporine derivative) and MLN518.
  • PLC412 N -benzoylstaurosporine
  • MLN518 MLN518
  • HSP90 inhibitors includes, but is not limited to, compounds targeting, decreasing or inhibiting the intrinsic ATPase activity of HSP90; degrading, targeting, decreasing or inhibiting the HSP90 client proteins via the ubiquitin proteosome pathway.
  • Compounds targeting, decreasing or inhibiting the intrinsic ATPase activity of HSP90 are especially compounds, proteins or antibodies which inhibit the ATPase activity of HSP90, e.g., 17-allylamino,17-demethoxygeldanamycin (17AAG), a geldanamycin derivative; other geldanamycin-related compounds; radicicol and HDAC inhibitors.
  • HDAC histone deacetylase
  • SAHA sodium butyrate and suberoylanilide hydroxamic acid
  • Specific HDAC inhibitors include MS275, SAHA, FK228 (formerly FR901228), Trichostatin A and compounds disclosed in U.S. Patent No.
  • Compounds which target, decrease or inhibit the activity/function of serine/theronine mTOR kinase are especially compounds, proteins or antibodies which target/inhibit members of the mTOR kinase family, e.g., RAD, RAD001 , CCI-779, ABT578, SAR543, rapamycin and derivatives/analogs thereof, AP23573 and AP23841 from Ariad, everolimus (CERTICAN ® ) and sirolimus.
  • somatostatin receptor antagonists refers to agents which target, treat or inhibit the somatostatin receptor, such as octreoride and SOM230.
  • Tuour cell damaging approaches refers to approaches, such as ionizing radiation.
  • ionizing radiation means ionizing radiation that occurs as either electromagnetic rays, such as X-rays and gamma rays; or particles, such as alpha and beta particles. Ionizing radiation is provided in, but not limited to, radiation therapy and is known in the art. See Hellman, Cancer, 4 th Edition, Vol. 1, Devita et al., Eds., pp. 248-275 (1993).
  • anti-leukaemic compounds includes, e.g., Ara-C, a pyrimidine analog, which is the 2'- ⁇ -hydroxy ribose (arabinoside) derivative of deoxycytidine. Also included is the purine analogue of hypoxanthine, 6-mercaptopurine (6-MP) and fludarabine phosphate.
  • EDG binders as used herein refers a class of immunosuppressants that modulates lymphocyte recirculation, such as FTY720.
  • CERTICAN ® (everolimus, FRAD) an investigational novel proliferation signal inhibitor that prevents proliferation of T-cells and vascular smooth muscle cells.
  • ribonucleotide reductase inhibitors refers to pyrimidine or purine nucleoside analogues including, but not limited to, fludarabine and/or ara-C; 6-thioguanine; 5-FU; cladribine; 6-mercaptopurine, especially in combination with ara-C against ALL; and/or pentostatin.
  • Ribonucleotide reductase inhibitors are especially hydroxyurea or 2-hydroxy- 7H-isoindole-1 ,3-dione derivatives, such as PL-1, PL-2, PL-3, PL-4, PL-5, PL-6, PL-7 or PL-8. See Nandy et al., Ada Oncologica, Vol. 33, No. 8, pp. 953-961 (1994). [76]
  • S-adenosylmethionine decarboxylase inhibitors includes, but is not limited to, the compounds disclosed in U.S. Patent No. 5,461 ,076.
  • VEGF vascular endothelial growth factor
  • WO 98/35958 e.g., 1-(4-chloroanilino)-4-(4-pyridylmethyl)phthalazine or a pharmaceutically acceptable salt thereof, e.g., the succinate, or in WO 00/09495, WO 00/27820, WO 00/59509, WO 98/11223, WO 00/27819 and EP 0 769 947; those as described by Prewett et al., Cancer Res, Vol. 59, pp. 5209-5218 (1999); Yuan et al., Proc NatlAcad Sci USA, Vol. 93, pp.
  • VEGF aptamer e.g., Macugon
  • FLT-4 inhibitors FLT-3 inhibitors
  • VEGFR-2 lgG1 antibody Angiozyme (RPI 4610)
  • Avastan ® Angiozyme
  • VEGF inhibitor compounds are compounds which target, decrease or inhibit the activity of VEGFR and are may be compounds, proteins or antibodies which inhibit or interact with at least one VEGF receptor tyrosine kinase, inhibit a VEGF receptor or bind to VEGF.
  • Photodynamic therapy refers to therapy which uses certain chemicals known as photosensitizing agents to treat or prevent cancers.
  • Examples of photodynamic therapy includes treatment with agents, such as, e.g., VISUDYNE ® and porfimer sodium.
  • Angiostatic steroids refers to agents which block or inhibit angiogenesis, such as, e.g., anecortave, triamcinolone, hydrocortisone, 11- ⁇ - epihydrocotisol, cortexolone, 17 ⁇ -hydroxyprogesterone, corticosterone, desoxycorticosterone, testosterone, estrone and dexamethasone.
  • Implants containing corticosteroids refers to agents, such as, e.g., fluocinolone and dexamethasone.
  • AT1 receptor antagonists include agents, such as DIOVAN ® .
  • ACE inhibitors include CIBACEN ® , benazepril, enazepril (LOTENSIN ® ), captopril, enalapril, fosinopril, lisinopril, moexipril, quinapril, ramipril, perindopril and trandolapril.
  • chemotherapeutic agents include, but are not limited to, plant alkaloids, hormonal agents and antagonists, biological response modifiers, preferably lymphokines or interferons, antisense oligonucleotides or oligonucleotide derivatives; or miscellaneous agents or agents with other or unknown mechanism of action.
  • references to the components (a) and (b) are meant to also include the pharmaceutically acceptable salts of any of the active substances.
  • active substances comprised by components (a) and/or (b) have, for example, at least one basic centre, they can form acid addition salts.
  • Corresponding acid addition salts can also be formed having, if desired, an additionally present basic centre.
  • Active substances having an acid group, e.g., COOH can form salts with bases.
  • the active substances comprised in components (a) and/or (b) or a pharmaceutically acceptable salts thereof may also be used in form of a hydrate or include other solvents used for crystallization.
  • one aspect of this invention is a method to predict response of a patient with a myeloid neoplasm, such as AML, to treatment with a therapeutic agent including an agent that can induce apoptosis or a tyrosine kinase inhibitor, i.e., an apoptosis inducing chemotherapy agent.
  • a therapeutic agent including an agent that can induce apoptosis or a tyrosine kinase inhibitor, i.e., an apoptosis inducing chemotherapy agent.
  • a sample of blood or bone marrow aspirate is obtained from the patient.
  • the sample of blood or bone marrow is then applied as a thin layer on a slide and dried and fixed, e.g., with methanol, neutral-buffered formalin or para-formaldehyde.
  • the degree of apoptosis of the blasts seen in the blood or bone marrow can then be determined by any method known in the art.
  • the likelihood of response of a patient with a myeloid neoplasm to an apoptosis inducing chemotherapy agent can be estimated from the pre treatment degree of apoptosis seen in the blasts in the peripheral blood or bone marrow sample.
  • the patient will be considered to be a responder to an apoptosis inducing chemotherapy agent.
  • the percentage of apoptosis in blast cells prior to treatment is greater than about 10%, the patient will be considered to be a responder to an apoptosis inducing chemotherapy agent.
  • the patient will be considered to be a responder to an apoptosis inducing chemotherapy agent, and in most preferred embodiments, if the percentage of apoptosis in blast cells prior to treatment is greater than about 30% or greater than 40%), the patient will be considered to be a responder to an apoptosis inducing chemotherapy agent.
  • myeloid neoplasm shall mean any neoplastic disorder of the myeloid lineage and includes but is not limited to AMLs, myelodysplastic syndromes and myelo-proliferative disorders. See Harris et al. (1999), supra; and Bennet et al., Hematol Oncol, Vol. 1 , pp. 187-192 (1982), both incorporated herein by reference in their entirety.
  • Apoptosis Assays One of skill in the art will know that apoptosis can be measured in many ways and, it is intended that any of these assays can be used in the methods of this invention. For general information on available apoptosis assays, see Apoptosis and Cell Proliferation, 2 nd Edition (Boehringer Mannheim GmbH, Biochemie 1998), incorporated by reference herein in its entirety.
  • the degree or extend of apoptosis is determined by means of a TUNEL assay.
  • the degree of apoptosis in the blasts in the blood or bone marrow from a patient with a myeloid neoplasm is determined prior to treatment with an apoptosis inducing chemotherapy agent. If the degree of apoptosis is less than about 5% the patient could be predicted to be a non-responder and an alternative form of treatment would be considered, e.g., a different type of chemotherapy agent or combination therapy with multiple agents.
  • the patient could be predicted to be a responder and treatment with the apoptosis inducing chemotherapy agent would proceed. If the degree of apoptosis is found to be between 5% and 20%, the patient could be predicted to be a partial responder and treatment may proceed, but other treatment including, but not limited to, combination therapy, will be considered.
  • the degree of apoptosis in the blasts in the blood or bone marrow from a patient with a myeloid neoplasm is determined prior to the patients inclusion in a study of an apoptosis inducing chemotherapy agent. If the degree of apoptosis is less than about 5%, the patient could be predicted to be a non-responder and will not be included in the study. If the degree of apoptosis is found to be about 20% or greater, such as for example greater than 30% or greater than 40%, the patient could be predicted to be a responder and would be included in the study with the apoptosis inducing chemotherapy agent. If the degree of apoptosis is found to be between 5% and 20%, the patient could be considered to be a partial responder and might or might not be included in the study depending on the individual circumstances.
  • Another aspect of the invention relates to the use of N-benzoylstaurosporine in the manufacture of a medicament for the treatment of myeloid neoplasms in a selected patient population, wherein the patient population is selected on the basis of a measurement of the pre-treatment extent of apoptosis in the blasts in a patient sample.
  • the patient population is selected if the degree of apoptosis is found to be about 20% or greater, such as for example greater than 30% or greater than 40%.
  • the patient population may be selected if the degree of apoptosis is found to be between 5% and 20%.
  • the patient sample comprises of a smear of blood, a smear of bone marrow aspirate or a cytospin of an aspirate isolated from said patient.
  • any method known in the art to measure the extent of apoptosis in the blasts or other cells of a myeloid neoplasm may be used in the methods of the invention. These methods may involve the use of various characteristics of the apoptosis process, as described below, and often make use of light microscopy to identify marked or tagged cells in the blood or bone marrow smear or slide. In order to establish a reliable percentage at least 10 cells should be counted, in a preferred embodiment at least 100 cells are counted. See Apoptosis and Cell Proliferation 2 nd Edition (1998), supra, incorporated by reference herein, in its entirety.
  • Methods for use in cell populations include assays for DNA fragments, such as the apoptotic DNA Ladder Kit [see Gavrieli, Sherman and Ben-Sasson, J Cell Biol, Vol. 119, No. 3, pp. 493-501 (1992)] and quantification of histone complex with DNA fragments with an ELISA.
  • assay for caspase activation can be used, including the detection of the cleavage of a substrate, such as poly ADP ribose polymerase (PARP).
  • PARP poly ADP ribose polymerase
  • Methods for use in individual cells include staining of chromosomal DNA after permeabilization, active labelling of cells by nick translation, the detection of translocated membrane components or the detection of damage or leakage of plasma membrane, such as by trypan blue or propidium iodide exclusion assays.
  • the apoptosis assay used is the TUNEL enzymatic labelling assay. See White et al., J Virol 52: 410 (1984).
  • the TUNEL assay makes use of the fact that extensive DNA degradation often occurs in the early stages of apoptosis. Cleavage of the DNA may yield double-stranded, LMW DNA fragments (mono- and olgionucleosomes), as well as single-strand breaks "nicks" in HMW-DNA. These DNA strand breaks can be detected by enzymatic labelling of the free 3'-OH termini with modified nucleotides, such as X-dUTP, X-brotin, DIG or fluorescein.
  • modified nucleotides such as X-dUTP, X-brotin, DIG or fluorescein.
  • TUNEL assay makes use of terminal dioxynucleotidyl transferase (TdT) to label the blunt ends of double- stranded DNA breaks independent of a template.
  • TUNEL refers to TdT-mediated X-dUTP nick end labelling.
  • Kits for performing any of the above assays can be obtained from Boehringer Mannheim, Indianapolis, IN.
  • Midostaurin is a staurosporine derivative, a tyrosine kinase inhibitor of a variety of targets, including but not limited to PKC, alpha, beta and gamma, VEGFR-2, c-kit, PDGFR- alpha, PDGFR-beta and FLT3.
  • Midostaurin has also been described as an apoptosis inducing chemotherapy agent able to induce apoptosis in malignant cells. (See, Fabbo, D Anticancer Drug Des. 15(1): 17-28 (February 2000).
  • blood samples are collected into an anticoagulant, e.g., EDTA.
  • the blood sample is then applied as a thin layer on a glass slide, dried and fixed with methanol, neutral-buffered formalin or para-formaldehyde.
  • the cells fixed on the slide are then assayed with a TUNEL apoptosis assay via the In Situ Cell Death Detection Kit (Boehringer Mannheim, Indianapolis, IN, USA) to determine the percentage of blast cells undergoing apoptosis. This result is correlated with the drug response data obtained by determining the percentage of blasts in the sample at the initiation of treatment and at the several time points after starting drug treatment. A substantial reduction in the percentage of blasts in the blood sample after initiation of the treatment is an indication of a positive clinical response to the study drug. [104] Results. Patient No.

Abstract

La présente invention a trait à des procédés de prédiction à partir de prélèvements sanguins ou de moelle osseuse quels patients avec des néoplasmes myéloïdes vont présenter une bonne réponse thérapeutique à des agents thérapeutiques qui opèrent, au moins en partie, par l'induction d'apoptose dans des cellules tumorales. Ces procédé peuvent contribuer à déterminer les choix de traitement et la sélection de patients pour des études sur des médicaments.
PCT/EP2004/013588 2003-12-01 2004-11-30 Procede de prediction de sensibilite aux medicaments dans des neoplasmes myeloides WO2005053704A1 (fr)

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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2006021456A1 (fr) * 2004-08-27 2006-03-02 Novartis Ag Combinaison thérapeutique impliquant une staurosporine et un inhibiteur du cytochrome cyp3a4
WO2019181465A1 (fr) 2018-03-19 2019-09-26 日鉄ケミカル＆マテリアル株式会社 Élément électroluminescent organique

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6077684A (en) * 1996-11-14 2000-06-20 Vanderbilt University Automated assay for measuring apoptosis in cell culture
WO2002046751A2 (fr) * 2000-11-13 2002-06-13 Vanderbilt University Methodes de prediction de reponse a une chimiotherapie
WO2003037347A1 (fr) * 2001-10-30 2003-05-08 Novartis Ag Derives de staurosporine inhibiteurs de l'activite tyrosine kinase du recepteur flt3

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6077684A (en) * 1996-11-14 2000-06-20 Vanderbilt University Automated assay for measuring apoptosis in cell culture
WO2002046751A2 (fr) * 2000-11-13 2002-06-13 Vanderbilt University Methodes de prediction de reponse a une chimiotherapie
WO2003037347A1 (fr) * 2001-10-30 2003-05-08 Novartis Ag Derives de staurosporine inhibiteurs de l'activite tyrosine kinase du recepteur flt3

Non-Patent Citations (6)

* Cited by examiner, † Cited by third party
Title
BLOOD, vol. 102, no. 11, 16 November 2003 (2003-11-16), 45TH ANNUAL MEETING OF THE AMERICAN SOCIETY OF HEMATOLOGY; SAN DIEGO, CA, USA; DECEMBER 06-09, 2003, pages 96a - 97a, ISSN: 0006-4971 *
DATABASE BIOSIS [online] BIOSCIENCES INFORMATION SERVICE, PHILADELPHIA, PA, US; 16 November 2003 (2003-11-16), BALI PURVA ET AL: "A combination of histone deacetylase inhibitor LAQ824 and the FLT-3 kinase inhibitor PKC412 is highly active against human AML cells with constitutively active mutant FLT-3 tyrosine kinase.", XP002317343, Database accession no. PREV200400172527 *
GROSIOS K: "Midostaurin, Novartis AG", CURRENT OPINION IN ONCOLOGIC, ENDOCRINE AND METABOLIC INVESTIGATIONAL DRUGS, CURRENT DRUGS, LONDON,, GB, vol. 2, no. 1, 2000, pages 92 - 103, XP008024624, ISSN: 1464-8466 *
KRAVTSOV ET AL: "Use of the microculture kinetic assay of apoptosis to determine chemosensitivities of leukemias", BLOOD, W.B. SAUNDERS, PHILADELPHIA, VA, US, vol. 92, no. 3, 1 August 1998 (1998-08-01), pages 968 - 980, XP002118802, ISSN: 0006-4971 *
KRAVTSOV V D ET AL: "CHEMOTHERAPY OF MYELOID LEUKEMIA DIRECTED BY A MICROCULTURE KINETIC (MICK)ASSAY FOR APOPTOSIS", BLOOD, W.B.SAUNDERS COMPANY, ORLANDO, FL, US, vol. 96, no. 11, PART 1, 16 November 2000 (2000-11-16), pages 724A, XP001106433, ISSN: 0006-4971 *
KRAVTSOV V D ET AL: "PREDICTION OF SURVIVAL AND RESPONSES TO CHEMOTHERAPY IN ACUTE MYELOGENEOUS LEUKEMIA (AML) BY THE MICROCULTURE KENETIC (MICK) ASSAY OF APOPTOSIS", BLOOD, W.B. SAUNDERS, PHILADELPHIA, VA, US, vol. 92, no. 10, 15 November 1998 (1998-11-15), pages 677A,AN2790, XP001106432, ISSN: 0006-4971 *

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2006021456A1 (fr) * 2004-08-27 2006-03-02 Novartis Ag Combinaison thérapeutique impliquant une staurosporine et un inhibiteur du cytochrome cyp3a4
WO2019181465A1 (fr) 2018-03-19 2019-09-26 日鉄ケミカル＆マテリアル株式会社 Élément électroluminescent organique

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