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WO1998003183A1 - Utilisation de 5,6-dihydro-5-azacytidine pour traiter le cancer de la prostate - Google Patents

Utilisation de 5,6-dihydro-5-azacytidine pour traiter le cancer de la prostate Download PDF

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Publication number
WO1998003183A1
WO1998003183A1 PCT/US1997/013102 US9713102W WO9803183A1 WO 1998003183 A1 WO1998003183 A1 WO 1998003183A1 US 9713102 W US9713102 W US 9713102W WO 9803183 A1 WO9803183 A1 WO 9803183A1
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Prior art keywords
azacytidine
cell
dihydro
administering
prostate cancer
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PCT/US1997/013102
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English (en)
Inventor
Daniel D. Von Hoff
Elzbieta Izbicka
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Ilex Oncology, Inc.
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Application filed by Ilex Oncology, Inc. filed Critical Ilex Oncology, Inc.
Priority to AU40461/97A priority Critical patent/AU4046197A/en
Publication of WO1998003183A1 publication Critical patent/WO1998003183A1/fr

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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
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/7042—Compounds having saccharide radicals and heterocyclic rings
    • A61K31/7052—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides
    • A61K31/706—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides containing six-membered rings with nitrogen as a ring hetero atom
    • A61K31/7064—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides containing six-membered rings with nitrogen as a ring hetero atom containing condensed or non-condensed pyrimidines
    • A61K31/7068—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides containing six-membered rings with nitrogen as a ring hetero atom containing condensed or non-condensed pyrimidines having oxo groups directly attached to the pyrimidine ring, e.g. cytidine, cytidylic acid
    • 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
    • A61K31/16—Amides, e.g. hydroxamic acids
    • A61K31/165—Amides, e.g. hydroxamic acids having aromatic rings, e.g. colchicine, atenolol, progabide
    • 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
    • A61K31/275—Nitriles; Isonitriles
    • 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
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/04—Peptides having up to 20 amino acids in a fully defined sequence; Derivatives thereof
    • A61K38/08—Peptides having 5 to 11 amino acids
    • A61K38/09—Luteinising hormone-releasing hormone [LHRH], i.e. Gonadotropin-releasing hormone [GnRH]; Related peptides
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
    • A61K45/06—Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca

Definitions

  • C-MT cytosine DNA methyltransferase
  • E-cadherin the "invasion-suppressor" gene is silenced by hypermethylation in human carcinomas and in human breast and prostate carcinomas (Yoshiura, K. et al. (1995), Proc. Natl. Acad. Sci. USA. 92:7416- 7419;Graff, L. R. et al. (1995), Cancer Res.. 55:5195-5199).
  • the latter study used azacytidine (azaC) to partially restore E-cad expression.
  • the present invention seeks to overcome deficiencies in the art by providing a treatment of prostate cancer that can cause the reversion of a hormone refractory prostate cancer cell to a hormone sensitive cell. Without limiting the present invention to a any single theoretical basis, it is contemplated that androgen independence of prostate cancer cells is associated with a lack of expression of the androgen receptor, possibly due to methylation of a gene control region mediated by cytosine-DNA-methyltransf erase (C-MT).
  • C-MT cytosine-DNA-methyltransf erase
  • the methods of the present invention utilize 5,6-dihydro-5-azacytidine, an inhibitor of C-MT, to restore expression of the androgen receptor, among other effects, and to cause the reversion of the cell to hormone sensitivity, thereby rendering the cell treatable with luteinizing hormone-releasing hormone (LHRH) agonists, androgen antagonists or other hormone manipulation.
  • LHRH luteinizing hormone-releasing hormone
  • An aspect of the present invention therefore, is a combination therapy using DHAC with conventional hormone therapy in the treatment of hormone refractory prostate cancer.
  • the present invention may be described as a method of increasing expression of the androgen receptor in a prostate cancer cell comprising contacting the cell with 5,6-dihydro-5-azacytidine under conditions effective to increase expression of the androgen receptor.
  • androgen receptor expression is increased in both androgen receptor negative and androgen receptor positive cells. Therefore, in light of this data, and in view of the other effects described herein, (increase in apoptosis and increased expression of E-cadherin), it is understood that treatment with DHAC may be beneficial for hormone refractory tumors and for hormone sensitive tumors as well.
  • Another aspect of the present invention is a method of increasing E- cadherin expression in a prostate cancer cell, comprising contacting the cell with 5,6-dihydro-5-azacytidine.
  • An increase in E-cadherin levels is an indication that a prostate tumor is expected to be more defined and less aggressive (better behaved). Therefore, an increase in E-cadherin as described herein is a further and important benefit of contacting a prostate tumor or cell with DHAC.
  • a surprising benefit of the disclosed methods is also an increase in apoptosis in prostate cancer cells upon contact with DHAC. Because prostate cancer represents, in some way, a failure of the apoptotic mechanism of the cell, an increase in apoptosis of the cancer cells is a promising benefit of the disclosed treatment methods.
  • the preferred cell to be contacted with DHAC is a human neoplastic prostate cell, and preferably the cell is contained in a subject and most preferably in a human prostate cancer patient.
  • An effective amount is described herein as an in vitro concentration of from a detectable level to about 10 ⁇ M 5,6-dihydro-5-azacytidine.
  • the in vitro level is that amount that is demonstrated to be effective in an in vitro embodiment, and is indicative of the level to be achieved at the site of the prostate cell in a subject.
  • the invention may be described as a method of treating a prostate tumor, including metathesis, in a subject comprising administering 5,6-dihydro-5-azacytidine to said subject in an amount effective to convert a hormone refractory tumor to a hormone sensitive tumor, or as a method of increasing E-cadherin expression in a prostate tumor in a subject, or even as a method of increasing apoptosis in prostate tumor cells in a subject.
  • the method comprises administering to a subject an effective amount of DHAC, wherein an effective amount is about 100 to about 2000 mg/m 2 per day. In certain preferred embodiments, this amount is administered for about 5 to 7 days, every three weeks to a month as determined by the practitioner. It is contemplated that the daily dosage may be higher in some cases, in particular when the duration of administering is shorter, even including up to 8 grams/m 2 in a single day. However, due to the side effects associated with DHAC, the preferred method is a lower dosage over a period of 5-7 days.
  • FIG. 1 shows the expression of the androgen receptor in LNCaP cells after seven days at various concentrations of DHAC.
  • FIG. 2 shows the expression of the androgen receptor in DU145 cells after seven days at various concentrations of DHAC.
  • FIG. 3 shows the expression of the androgen receptor in MPC3 cells after seven days at various concentrations of DHAC.
  • FIG. 4 shows C-MT activity in LNCap cells after treatment at various concentrations of DHAC.
  • FIG. 5 shows C-MT activity in DU145 cells after treatment at various concentrations of DHAC.
  • FIG. 6 shows the levels of apoptosis in DU145 cells (TdT assay) at 1, 3 and 5 days of culture after treatment with DHAC.
  • FIG. 7 shows the levels of apoptosis in LNCap cells (TdT assay) at 1 , 3 and 5 days of culture after treatment with DHAC.
  • FIG. 8 shows cell growth in DU145 cells after treatment with DHAC.
  • FIG. 9 shows cell growth in LNCap cells after treatment with DHAC.
  • FIG. 10 shows upregulation of androgen receptor expression in all four tested lines of human prostate cells.
  • FIG. 1 1 shows the regulation of AR gene expression and C-MT activity by DHAC in TSUPrl prostate carcinoma.
  • the present invention may be described in a broad aspect as a method of treating prostate cancer by the administration of 5,6-dihydro-5-azacytidine.
  • This treatment is shown herein to have multiple effects that are contemplated to be beneficial to a prostate tumor patient.
  • One such effect is the upregulation of the androgen receptor protein. It is the down-regulation of androgen receptor that is thought to be the cause of hormone refractory tumors, which do not respond to hormonal manipulation.
  • the increase in expression of the androgen receptor caused by contacting those cells with 5, 6-dihydro-5 -azacytidine is expected to result in a reversion of a hormone refractory prostate tumor to hormone sensitivity.
  • Results obtained and disclosed herein with prostate cancer cell lines that are both androgen receptor positive and negative demonstrate the ability of this method to increase androgen receptor expression.
  • Other broad aspects of the present invention include methods of increasing apoptosis in prostate cancer cells by contacting those cells with DHAC. This surprising result is an important embodiment of the present invention, particularly if those results are tumor cell specific.
  • An important embodiment of the present invention is a method of increasing expression of E-cadherin in a prostate cancer cell by contacting the cell with DHAC.
  • E-cadherin (E-cad) is reported to suppress tumor cell invasion and metastasis in experimental tumor models and E-cad expression is low in poorly differentiated, advanced stage carcinomas (Graff, L. R. et al. (1995), Cancer Res.. 55:5195-5199). This loss of expression may be due to hypermethylation of the E-cad gene promoter region, and therefore, administration of DHAC may have the effect of hypomethylation of that region, resulting in increased expression of E-cad and a less aggressive tumor.
  • C-MT cytosine DNA methyltransferase
  • Various genes are upregulated or down-regulated as part of the process of conversion of a normal cell to a cancer cell, and DHAC is shown herein to be effective in altering the expression of various cancer genes, possibly by affecting the activity of C-MT.
  • a further aspect of the present invention is a method of causing the reversion of hormone insensitive prostate tumors and metastases to hormone sensitivity, thus allowing the tumor to be treated or controlled by conventional hormone manipulation therapy.
  • Hormone therapies useful for treating prostate cancer include, orchiectomy (castration) and treatment with agonists of lutenizing hormone- releasing hormone (LHRH) (e.g. leuprolide acetate or goserelin acetate). Such therapies may additionally be augmented by administration of an agent which inhibits the action of androgens (e.g. bicalutamide or flutamide).
  • LHRH lutenizing hormone- releasing hormone
  • Such therapies may additionally be augmented by administration of an agent which inhibits the action of androgens (e.g. bicalutamide or flutamide).
  • the invention also provides a method for treating prostate cancer in a mammal (e.g. a human), comprising administering 5,6-dihydro-5-azacytidine or a pharmaceutically acceptable salt thereof, in combination with hormone therapy.
  • a mammal e.g. a human
  • the invention also provides a method for treating prostate cancer in a mammal (e.g.
  • a human comprising administering 5,6-dihydro-5-azacytidine or a pharmaceutically acceptable salt thereof, in combination with an agent that inhibits the action of androgens.
  • an agent that inhibits the action of androgens an agent that inhibits the action of androgens.
  • the individual agents including 5,6-dihydro-5-azacytidine or a pharmaceutically acceptable salt thereof
  • the present disclosure includes examples in which four human prostate carcinoma cell lines (LNCaP, DU145, MPC3, and TSUPrl) were incubated for a week with 5 ⁇ M azaC or 5 ⁇ M 5,6-dihydro-5-azacytidine (DHAC).
  • LNCaP human prostate carcinoma cell lines
  • DU145 human prostate carcinoma cell lines
  • MPC3 DU145
  • MPC3 DU145
  • MPC3 5 ⁇ M 5,6-dihydro-5-azacytidine
  • DHAC 5,6-dihydro-5-azacytidine
  • TSUprl express low levels of the AR receptor
  • LNCaP expresses high levels of the receptor.
  • the cells are treated in culture with two types of C-MT inhibitors: 5-azacytidine (azaC) and 5,6-dihydro-5-azacytidine (DHAC). To determine the optimal conditions for treatment, the cells are cultured in the presence of 10% fetal calf serum for 3, 5, and 7 days with a single dose of
  • DHAC or azaC (0.1, 0.5, 2.5, and 10 ⁇ M).
  • DHAC or azaC (0.1 , 0.5, 2.5, and 10 ⁇ M) is freshly added to the cells every 2 days for a total of 8 days. Control groups in each cell line have no inhibitor added.
  • the cells are harvested and cell pellets are collected by centrifugation and several assays are performed. AR protein expression may be determined by a ligand competition assay. The pellet is solubilized and the cytosolic fraction is isolated by ultracentrifugation. The cytosol is incubated with 3 H dihydrotestosterone (DHT) in the presence of excess unlabeled DHT to determine non-specific binding. The amount of specifically bound DHT is measured by liquid scintillation counting.
  • DHT dihydrotestosterone
  • AR gene expression is determined by RT PCR and northern blots.
  • Messenger RNA is extracted from the pellet using Fast Trac (Stratagene) reagent.
  • Complementary DNA cDNA
  • PCR ® polymerase chain reaction
  • Positive controls ⁇ -actin
  • negative controls are included.
  • Northern blots are hybridized with 32 P labeled cDNA probes for the AR. Quantitation of the binding is done by video imaging.
  • Cytosine-DNA methyltransferase (C-MT activity is also assayed.
  • the pellet is lysed in the presence of protease inhibitors and treated with RNase to deplete RNA, which inhibits C-MT activity.
  • the C-MT activity is determined using poly(dI/dC) and 3 H S-adenosyl methionine as the substrates, as described by Issa and colleagues (Issa, J. P. et al. (1993), J. Natl. Cancer Inst.. 85: 1235- 1240).
  • DHAC and azaC affected the expression of other genes
  • DU145 and LNCaP cell lysates which had previously tested positively for increased expression of the AR, were electrophoresed in polyacrylamide gels and transblotted to nitrocellulose.
  • the blots were developed with rabbit anti-E- cadherin polyclonal antibody.
  • the results showed that DHAC and azaC increased the expression of E-cadherin.
  • DHAC This agent and other C-MT inhibitors like DHAC, in addition to increasing the expression of the AR gene, might also help in restoring the expression of "invasion-suppressor" genes such as E-cadherin. Therefore, the benefits of restored expression of the AR gene would be enhanced by the restoration of the cell's natural defense mechanisms.
  • DHAC or 4-amino-3,6-dihydro-l- ⁇ -D-ribofuranosyl-l,3,5-triazin-2(lH)- ine-monohydrochloride was developed as an alternative to 5-azacytidine, an antimetabolite with demonstrated antitumor activity in children and adults with acute non-lymphocytic leukemia.
  • a method of preparing DHAC from 5- azacytidine by reduction of the 5,6-double bond of 5-azacytidine is described in U.S. Patent 4,058,602 (incorporated herein in its entirety by reference).
  • DHAC demonstrated similar in vitro and in vivo activity to that of 5 -azacytidine although higher doses are required.
  • 5-azacytidine The major dose limiting toxicity of 5- azacytidine given as a bolus dose is nausea and vomiting which can be overcome by a five day continuous infusion.
  • 5-azacytidine is typically administered intravenously from 50 to 400 mg/m 2 /day for five days at intervals of 2 to 3 weeks (Remington's Pharmaceutical Sciences, 18th edition, 1990, Mack Publishing Company, Easton, PA, 18042 pi 158).
  • 5-azacytidine is unstable in aqueous solution. Therefore, Beisler synthesized DHAC to overcome 5-azacytidine's water instability and to allow for continuous infusion without concern for drug hydrolysis in aqueous formulation (Beisler, J. A. et al. (1 77), J. Med. Chem.. 20:806-812).
  • 5,6-Dihydro-5-azacytidine is normally available as a white lyophilized powder.
  • Intact vials are stable for at least 4 years at room temperature or at 4°C.
  • a 500 mg vial may be reconstituted with 9.6 ml of Sterile Water for Injection, USP. This results in a clear solution containing 50 mg of DHAC and 30 mg of mannitol at a pH of 3 to 5.
  • the drug may be diluted in multiple types of intravenous infusion solutions.
  • the reconstituted solutions are stable for at least 48 hours at room temperature.
  • DHAC is also stable for 48 hours when diluted into 5% dextrose, 0.9% sodium chloride and Lactated Ringer's Injection, USP.
  • DHAC is not a prodrug of 5 -azacytidine, but is incorporated into nuclear RNA. It inhibits methylation of ribosomal and transfer RNA. It also inhibits transcription of ribosomal RNA and nuclear RNA. The overall effect is a decrease in the synthesis of methylated bases into RNA and a decrease in protein synthesis. DHAC can also substantially reduce DNA methylation levels. Accordingly, in addition to being useful for treating prostate cancer, DHAC may be useful for the treatment of other diseases wherein hypermethylation is implicated. Such diseases include but are not limited to thalassemia major and myodisplastic syndrom. The following examples are included to demonstrate preferred embodiments of the invention.
  • DHAC 5,6-dihydro-5-azacytidine upregulated the expression of the androgen receptor (AR) in LNCaP (FIG. 1) and DU145 (FIG. 2) cells.
  • AR androgen receptor
  • the expression of AR increased in DU145 cells from an undetectable level to 1.5 femtomoles/mg cytosol protein.
  • MPC3 cells was not significantly affected (FIG. 3).
  • C-MT activity was strongly inhibited by DHAC in LNCaP cells (80% inhibition ) (FIG. 4) and was apparently increased in DU145 cells (FIG. 5).
  • DHAC 5,6-dihydro-5-azacytidine hydrochloride
  • Pellets from prostate cells were homogenized on ice in chilled 10 mM Tris pll 8, 1.5 mM EDTA, 20 mM sodium molybdate, 10% glycerol, and 5 mM DTT (TEM 20 DG) buffer in a glass-Teflon homogenizer with a motor-driven pestle at a proportion of 3 ml buffer per 0.1 ml packed cells.
  • the homogenate was centrifuged at 40,000 rpm for 30 minutes, the supernatant containing cytosol proteins was incubated with pellet from a half volume of dextran-coated charcoal (DCC) for 10 minutes, and centrifuged 10 minutes at 2,000xg.
  • DCC dextran-coated charcoal
  • Cvtosine-DNA methvltransferase fC-MTI activity assay The cytosine-DNA methyltransferase (C-MT) activity assay was done according to the modified method described in Issa, J. P. et al. (1993), J. Natl. Cancer Inst.. 85:1235-1240).
  • Human prostate cell pellets were homogenized with disposable plastic pestles in 1.5 ml Eppendorf tubes containing 500 ⁇ l of 50 mM Tris (pH 7.8), 1 mM EDTA, 0.1% sodium azide, 10% glycerol, 1% Tween 80, 1 mM dithiothreitol, 6 mg/ml phenylmethyl sulfonyl fluoride, and 100 ⁇ g/ l Rnase A (lysis buffer). The homogenate was passed several times through a 27 gauge needle and aliquots were removed for protein measurement by measuring the absorbance of the samples at 280 nm (A 2g0 ) against the lysis buffer.
  • DNA C-MTase activity determination 5 ⁇ g of the cellular protein lysate was mixed with 0.5 ⁇ g of a synthetic DNA polymer, poly (d(I-C) (Pharmacia Biotech, Piscataway, NJ) and 3 ⁇ Ci S-adenosyl methionine (80 Ci/mmol) in a total volume of 20 ⁇ l and incubated at 37°C for 2.5 hours.
  • the reaction was stopped by adding 300 ⁇ l of 1% aminosalicylate, 5% butanol, 125 mM NaCl, 0.25 mg/ml salmon sperm DNA, and 1 mg/ml proteinase K. The samples were vortexed briefly and incubated another 30 minutes at 37°C.
  • DNA was purified by phenol-chloroform extraction and ethanol precipitation. Samples were placed at -70°C for 15 minutes and centrifuged at 12,000xg at 4°C for 5 minutes. The pellet was washed twice in 1 ml ice cold 70% ethanol, and air dried. RNA was removed by resuspension in 25 ⁇ l of 0.3 M NaOH and incubation at 37°C for 30 minutes.
  • the terminal deoxynucleotidyl transferase assay detects apoptotic cells by direct fluorescence detection of digoxigenin-labeled genomic DNA.
  • the assay the addition of digoxigenin-11-dUTP and dATP to DNA is catalyzed by TdT.
  • the heteropolymer product is detected with fluorescein-labeled anti- digoxigenin antibody.
  • an intense emission signal is generated at 523 nm.
  • the cells were harvested, counted, then centrifuged and incubated for 10 minutes in 4% formalin. The cells were then spread on silanized slides and air dried. The slides were washed in PBS, and incubated with TdT and the digoxigenin-labeled substrates (Oncor-ApopTag kit) and incubated for 1 hour at 37°C. After incubation the cells were washed in a stop/wash buffer for 30 minutes and then antidigoxigenin-fluorescein was added for 30 minutes. The cells were counterstained with propidium iodide/Vectashield and viewed under a fluorescence microscope. A minimum of 500 cells were counted and the percentage of apoptotic cells was calculated.
  • Example 2 For the in vivo studies, nude mice are implanted with human prostate cancer cells that express low and intermediate [AR(-)], or high levels [AR(+)] of the androgen receptor gene. The animals are treated with variable doses of cytosine DNA methyltransferase (C-MT) inhibitors and varying schedules of administration and exposure to the agents, to determine the effects of C-MT inhibitors on the expression of AR in primary and metastatic tumor sites, tumor burden, number of metastases, body weight and animal survival.
  • C-MT cytosine DNA methyltransferase
  • the preferred inhibitor is 5,6-dihydro-5-azacytidine (DHAC), which may be compared to the more toxic 5-azacytidine.
  • DHAC 5,6-dihydro-5-azacytidine
  • the drug's ability to increase expression of the androgen receptor in the Dunning Mat Ly-Lu rat prostate cancer line is tested. These cancer cells are androgen receptor negative and highly aggressive.
  • the parent rat prostate cancer cell line, HI-F, which is androgen insensitive but has detectable levels of androgen receptors is also tested. When treatment with drug increases androgen receptor levels, the effect on animal survival is tested by hormone ablative therapy (surgical castration).
  • DHAC has been used in a mouse myeloma model at a dose of 200 mg/kg.
  • the dosage in the present study is optimized as follows: Eighteen animals are inoculated with tumor and randomized into three groups of six to receive 100 mg/kg or 200 mg/kg IP in a single daily dose for five days. The third group receives no drug. Two animals from each group are sacrificed at 7 days, 14 days and 21 days. Tumors are harvested and androgen receptor levels are measured.
  • 80 animals are injected with tumor cells into the hind limb and randomly assigned to 4 groups of 20 as follows: a) untreated animals b) untreated animals castrated on day 10 c) DHAC treated animals d) DHAC treated animals castrated on day 10.
  • Treated animals are given a daily dose of DHAC beginning on day zero to day four, for a total of five doses. Animals are closely monitored for tumor size, and if large volumes (>20 cc) occur, the animals are sacrificed. Animals are followed until all untreated animals are sacrificed. Tumors are harvested in all groups for receptor analysis. If treated, castrated animals show decrease in tumor size and prolonged survival, they are maintained until there is a statistical difference in survival curves over the other groups, at which time all surviving animals are sacrificed. The protocol is done first in the aggressive Mat Ly-Lu cell line, followed by an identical study in the Hi-F line.
  • Example 3 Clinical trials of DHAC in the treatment of prostate cancer are designed and conducted as follows.
  • Patients with AR(-) biopsy-proven prostate cancer which is refractory to hormonal treatment are treated with DHAC and then re-biopsied to determine the AR status of their tumors. If their tumors are converted to the AR(-) state, the patients are treated with hormonal manipulation therapy and observed for response to that therapy.
  • DHAC may be administered on a monthly or three week schedule as a continuous infusion of 5 to 8 g/m 2 given in 24 hours, or infused over a period of up to 5 days at about 100 to about 2000 mg/m 2 per day in 5% dextrose and possibly 0.9% NaCl in water.
  • Patients may also be administered narcotic analgesics and/or nonsteroidal anti-inflammatory agents if pain is a problem (Carr, B. I. et al. (1987), Cancer Research. 47:4199-4201 ; Creagan, E. T. et al. (1993), Am J. Clin. Oncol.. 16(3):243-244). It is also understood that the dosage and frequency may be adjusted according to patient tolerance of the drug as well as disease response.

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Abstract

Procédé de traitement du cancer de la prostate, qui consiste à administrer une quantité efficace de 5,6-dihydro-5-azacytidine, ou un sel pharmaceutiquement acceptable de ladite substance, soit seul, soit en combinaison avec une thérapie hormonale. La présente invention concerne un procédé permettant d'augmenter l'expression du récepteur d'androgène dans une cellule cancéreuse prostatique, un procédé permettant d'augmenter l'expression de E-cadhérine dans une cellule cancéreuse prostatique et un procédé permettant d'induire l'apoptose dans une cellule prostatique.
PCT/US1997/013102 1996-07-22 1997-07-22 Utilisation de 5,6-dihydro-5-azacytidine pour traiter le cancer de la prostate WO1998003183A1 (fr)

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Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4058602A (en) * 1976-08-09 1977-11-15 The United States Of America As Represented By The Department Of Health, Education And Welfare Synthesis, structure, and antitumor activity of 5,6-dihydro-5-azacytidine
EP0417454A2 (fr) * 1989-08-07 1991-03-20 Abbott Laboratories Analogues de LHRH à dimension réduite
US5162117A (en) * 1991-11-22 1992-11-10 Schering Corporation Controlled release flutamide composition

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4058602A (en) * 1976-08-09 1977-11-15 The United States Of America As Represented By The Department Of Health, Education And Welfare Synthesis, structure, and antitumor activity of 5,6-dihydro-5-azacytidine
EP0417454A2 (fr) * 1989-08-07 1991-03-20 Abbott Laboratories Analogues de LHRH à dimension réduite
US5162117A (en) * 1991-11-22 1992-11-10 Schering Corporation Controlled release flutamide composition

Non-Patent Citations (3)

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