WO2000059518A2 - Composition and method for inducing apoptosis in prostate cancer cells - Google Patents
Composition and method for inducing apoptosis in prostate cancer cells Download PDFInfo
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- WO2000059518A2 WO2000059518A2 PCT/CA2000/000342 CA0000342W WO0059518A2 WO 2000059518 A2 WO2000059518 A2 WO 2000059518A2 CA 0000342 W CA0000342 W CA 0000342W WO 0059518 A2 WO0059518 A2 WO 0059518A2
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- A61K31/7088—Compounds having three or more nucleosides or nucleotides
- A61K31/711—Natural deoxyribonucleic acids, i.e. containing only 2'-deoxyriboses attached to adenine, guanine, cytosine or thymine and having 3'-5' phosphodiester links
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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
- A61K31/7088—Compounds having three or more nucleosides or nucleotides
- A61K31/713—Double-stranded nucleic acids or oligonucleotides
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/66—Microorganisms or materials therefrom
- A61K35/74—Bacteria
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
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- A61K9/5068—Cell membranes or bacterial membranes enclosing drugs
Definitions
- the present invention relates to a method for treating prostate cancer.
- Cancer is an aberrant net accumulation of atypical cells, which can result from an excess of proliferation, an insufficiency of apoptosis, or a combination of the two.
- Apoptosis is an active cellular death process characterized by distinctive morphological changes that include condensation of nuclear chromatin, cell shrinkage, nuclear disintegration, plasma membrane blebbing, and the formation of membrane-bound apoptotic bodies (Wyllie et al. Int. Rev. Cytol. 68:251, 1980).
- a molecular hallmark of apoptosis is degradation of the cell's nuclear DNA into oligonucleosomal-length fragments as the result of activation of endogenous endonucleases (Wyllie A.H. Nature 284:555, 1981).
- Prostate cancer is a major cause of cancer deaths among the male population.
- radical prostatectomy, radiotherapy, a combination of the two, and brachytherapy with radiolabeled seed implants are the most common treatments.
- Radical prostatectomy can result in impotence and incontinence.
- Radiotherapy is associated with prostate specific antigen (PSA) recurrence.
- Brachytherapy can be used only in carefully selected patients.
- Androgen ablation for use in patients with androgen-dependent prostate cancer, includes surgical castration (bilateral orchiectomy) and chemical castration using anti-androgens and leutinizing hormone production suppressors. Many patients refuse orchiectomy. Anti-androgens and leutinizing hormone production suppressors, used together to effect total androgen blockade, result in androgen withdrawal symptoms and, also, are very expensive.
- the response to androgen ablation therapy is finite and lasts a medium of 12 to
- p53 gene mutation which results in the accumulation of non-functional p53 protein, is known to be associated with a lack of responsiveness to androgen ablation therapy in the treatment of prostate cancer (Navone et al., J. Natl. Cancer Inst. 85:1657, 1993).
- Therapeutic agents that are effective in inhibiting the proliferation of and inducing apoptosis in cancer cells in vitro are recognized as having therapeutic utility for the in vivo treatment of cancer (Furukawa et al., J. Surg. Oncol. 48:188, 1991).
- the anti-proliferative activity in vitro of chemotherapeutic agents towards prostate adenocarcinoma cancer cells has been shown to positively correlate with their in vivo anti-cancer activity (Pienta et al., Prostate 26:270, 1995).
- the activity of many of these therapeutic agents is however influenced by p53 mutation status, and those agents whose mechanism of action appears to be independent of p53 are non-selective and highly toxic.
- Chemotherapy for use in patients with androgen-independent (hormone refractory) prostate cancer, has palliation as its primary goal.
- the toxic side effects of chemotherapy are debilitating, often dose limiting, and compromise the quality of the patient's life.
- Castration or androgen ablation therapy induces apoptosis in prostate tumors.
- the amount of apoptosis occurring in prostate tumors after androgen ablation has been correlated with p53 status (Westin et al., Am. J. Pathol. 146:1368, 1995).
- p53 status Westin et al., Am. J. Pathol. 146:1368, 1995.
- almost all patients with metastatic prostate cancer will escape first-line androgen ablation therapy.
- the present invention satisfies the above need by providing a method for treating prostate cancer in an animal, including a human, wherein a composition comprising Mycobacterium phlei (M. phlei) DNA (M-DNA) and M-DNA, wherein the M-DNA is preserved and is complexed with the M. phlei cell wall (MCC), is administered to the animal in need of such treatment in an amount effective to have an anti-neoplastic effect on cancer cells in the prostate of the animal.
- MTC Mycobacterium phlei
- M-DNA and MCC are simple and relatively inexpensive to prepare, their activities are reproducible among preparations, they remain therapeutically stable over time, and they are effective at dose regimens that are associated with minimal toxicity even upon repeated administration.
- M. phlei are grown in liquid medium and harvested.
- the M. phlei are disrupted, and the solid components of the disrupted M. phlei are collected by centrifugal sedimentation.
- the solid components are deproteinized, delipidated, and washed.
- M-DNA is purified from MCC or is purified directly from M. phlei. DNase-free reagents are used to minimize DNA degradation (preserve DNA) during preparation of
- MCC and M-DNA are administered to an animal, including a human, having prostate cancer in an amount effective to prevent, treat or eliminate prostate cancer cells in the animal, including the human, having prostate cancer.
- additional therapeutic agents including, but not limited to, anti-androgens, chemotherapeutic agents, immunomodulatory agents and steroids can be administered with the M-DNA and MCC.
- M-DNA and of MCC to prevent, treat or eliminate prostate cancer cells addresses a long felt unfulfilled need in the medical arts and provides an important benefit for animals, including humans.
- an object of the present invention is to provide a composition and method effective to prevent prostate cancer.
- Another object of the present invention is to provide a composition and method effective to prevent hormone sensitive prostate cancer.
- Another object of the present invention is to provide a composition and method effective to prevent hormone insensitive prostate cancer. Another object of the present invention is to provide a method effective to treat prostate cancer.
- Another object of the present invention is to provide a method effective to treat hormone sensitive prostate cancer.
- Another object of the present invention is to provide a method effective to treat hormone insensitive prostate cancer.
- Another object of the present invention is to provide a method effective to eliminate prostate cancer.
- Another object of the present invention is to provide a method that has an anti- neoplastic effect on prostate cancer cells. Another object of the present invention is to provide a method that has an anti- neoplastic effect on hormone-sensitive prostate cancer cells.
- Another object of the present invention is to provide a method effective to inhibit proliferation of prostate cancer cells.
- Another object of the present invention is to provide a method effective to inhibit proliferation of hormone sensitive prostate cancer cells.
- Another object of the present invention is to provide a method effective to inhibit proliferation of hormone insensitive prostate cancer cells.
- Another object of the present invention is to provide a method effective to induce apoptosis in hormone insensitive prostate cancer cells.
- Another object of the present invention is to provide a method effective to potentiate the activity of other therapeutic agents in the treatment of prostate cancer.
- Another object of the present invention is to provide a method effective to potentiate the anti-neoplastic effect of anti-androgenic agents in the treatment of prostate cancer.
- Another object of the present invention is to provide a method effective to potentiate the anti-neoplastic effect of chemotherapeutic agents in the treatment of prostate cancer.
- Another object of the present invention is to provide a method effective to potentiate the anti-neoplastic effect of radiation in the treatment of prostate cancer.
- Another object of the present invention is to provide a method that maintains its effectiveness over time.
- FIG. 1 Inhibition of proliferation of human prostate cancer cells by MCC.
- FIG. 2 Inhibition of proliferation of LNCaP and PC3 human prostate cancer cells by M-DNA.
- FIG. 3 Inhibition of proliferation of PC3 human prostate cancer cells by MCC and by DNase I treated MCC.
- FIG. 4 Morphological changes in PC3 human prostate cancer cells after treatment with MCC.
- FIG. 5 Induction of DNA fragmentation in PC3 human prostate cancer cells by MCC.
- FIG. 6 Release of NuMA from (A) LNCaP and (B) PC3 human prostate cancer cells by MCC. DETAILED DESCRIPTION OF THE INVENTION
- the present invention provides a method for treating prostate cancer in an animal, including a human, wherein a composition comprising a mycobacterial DNA (B-DNA) and B-DNA, wherein the B-DNA is preserved and complexed on a mycobacterial cell wall (BCC), is administered to the animal in need of such treatment in an amount effective to have an anti-neoplastic effect on cancer cells in the prostate of the animal.
- a composition comprising a mycobacterial DNA (B-DNA) and B-DNA, wherein the B-DNA is preserved and complexed on a mycobacterial cell wall (BCC)
- BCC mycobacterial cell wall
- the present invention provides a method for treating prostate cancer in an animal, including a human, wherein a composition comprising Mycobacterium phlei
- M. phlei DNA M-DNA
- MCC M. phlei cell wall
- the phrase “complexed on” refers to the physical association of M-DNA with M. phlei cell wall.
- response refers to inhibition of proliferation of cancer cells, induction of apoptosis in cancer cells, and stimulation of bioactive molecule production by immune system cells.
- bioactive molecules refers to cytokines and reactive oxygen species.
- immune system cells refers to macrophages, monocytes, leukocytes, T-cells, B-cells, NK cells, dendritic cells, Langerhan cells, interstitial cells and support cells.
- bacterial species can be used to practice the present invention including, but not limited to, Coryneform species, Corynebacterium species, Rhodococcus species, Eubacterium species, Bordetella species, Escherichia species, Listeria species, Nocardia species and Mycobacterium species.
- a Mycobacterium species is used including, but not limited to, M. smegmatis, M. fortuitum, M. kansaii, M. tuberculosis, M. bovis, M. vaccae, M. avium and M. phlei. Most preferably, the Mycobacterium species M. phlei is used.
- Methods to increase anti-neoplastic activity include, but are not limited to, chemically supplementing or biotechnologically amplifying the M-DNA and MCC with stimulatory sequences or confirmations of DNA derived from the same or different bacterial species; supplementing the M-DNA and MCC with naturally occurring or chemically-synthesized nucleic acids; and, complexing the M-DNA and MCC to natural or synthetic carriers.
- therapeutic agents including, but not limited to, anti-androgenic, chemotherapeutic, steroidal and immunological agents can be included in the M-DNA and
- MCC composition of the present invention M-DNA or MCC and an optional therapeutic agent can be administered simultaneously or sequentially on the same or different schedules.
- Anti-androgenic agents include, but are not limited to, flutamide, bicalutamide, nilutamide, megestrol acetate, adrenocorticotropic hormone secretion inhibitors, ketoconazole, estrogens, anti-estrogens and LHRH production suppressors.
- Chemotherapeutic agents include, but are not limited to, anti-metabolites, DNA damaging, microtubule destabilizing, microtubule stabilizing, actin depolymerizing, growth inhibiting, topoisomerase inhibiting, HMG-CoA inhibiting, purine inhibiting, pyrimidine inhibiting, metaloproteinase inhibiting, CDK inhibiting, caspase inhibiting, proteaosome inhibiting, angiogenesis inhibiting, differentiation inducing and immunotherapeutic drugs.
- agents include, but are not limited to, anthracycline antibiotics such as doxorubicin and mitoxantrone, estramustine, vinblastine, paclitaxel, etoposide, cyclophosphamide, cisplatin, carboplatin and combinations of the above with or without the addition of steroid drugs.
- anthracycline antibiotics such as doxorubicin and mitoxantrone, estramustine, vinblastine, paclitaxel, paclitaxel, etoposide, cyclophosphamide, cisplatin, carboplatin and combinations of the above with or without the addition of steroid drugs.
- Immunological agents include, but are not limited to, cytokines, chemokines, interferons, interleukins, polyclonal antibodies and monoclonal antibodies.
- compositions comprising M-DNA and MCC and a pharmaceutically acceptable carrier are prepared by uniformly and intimately bringing into association the M-DNA and the MCC with liquid carriers, with solid carriers or with both.
- Liquid carriers include, but are not limited to, aqueous carriers, non-aqueous carriers or both.
- Solid carriers include, but are not limited to, biological carriers, chemical carriers or both.
- M-DNA and MCC can be administered in aqueous suspension, oil emulsion, water in oil emulsion, water-in-oil-in-water emulsion, site-specific emulsion, long-residence emulsion, sticky-emulsion, microemulsion, nanoemulsion, liposomes, microparticles, microspheres, nanospheres, nanoparticles, minipumps, and with various natural or synthetic polymers that allow for sustained release.
- M-DNA and MCC can be used with any one, all, or any combination of excipients regardless of the carrier used to present the composition to the responding cells.
- excipients include, but are not limited to, anti-oxidants, buffers, bacteriostats, suspending agents, surfactants and tensio active agents.
- MCC is administered as an aqueous suspension, wherein the size range of the MCC is preferably about 0.025 to 1.000 ⁇ m, more preferably from about 0.050 to 0.750 ⁇ m and most preferably from about 0.100 to 0.500 ⁇ m.
- the MCC is suspended in a pharmaceutically acceptable carrier such as, but not limited to, DNase-free water, saline or phosphate buffered saline (PBS) and is homogenized and submicronized by standard procedure such as, but not limited to, sonication and microfluidization.
- a pharmaceutically acceptable carrier such as, but not limited to, DNase-free water, saline or phosphate buffered saline (PBS) and is homogenized and submicronized by standard procedure such as, but not limited to, sonication and microfluidization.
- lyophilized MCC is suspended in sterile water and sonicated at 20% output for 5 min (Model W-385 Sonicator, Heat Systems-Ultrasonics Inc) or microfluidized at 15,000- 30,000 psi for one or more flow-throughs (Model M-110Y; Microfluidics, Newton, MA).
- the mixture is either aseptically processed or terminally sterilized.
- An optional therapeutic agent or stabilizing agent can be added to the MCC during submicronization or homogenization or before or after sterilization.
- M-DNA is administrated as an aqueous suspension in the size range preferably from about 2 to > 12 000 b.p, more preferably from about 2 to 250 b.p. and most preferably from about 2 to 20 b.p.
- the M-DNA is suspended in a pharmaceutically acceptable carrier such as DNase-free water and is homogenized and fragmented by standard procedures such as, but not limited to, sonication and microfluidization.
- the mixture is aseptically processed or terminally sterilized.
- An optional therapeutic agent or stabilizing agent can be added to the M-DNA during sonication or homogenization or before or after sterilization
- M-DNA or MCC are emulsified with a mineral oil or with a neutral oil such as, but not limited to, a diglyceride, a triglyceride, a phospholipid, a lipid, an oil and mixtures thereof, wherein the oil contains an appropriate mix of polyunsaturated and saturated fatty acids.
- a neutral oil such as, but not limited to, a diglyceride, a triglyceride, a phospholipid, a lipid, an oil and mixtures thereof, wherein the oil contains an appropriate mix of polyunsaturated and saturated fatty acids.
- examples include, but are not limited to, soybean oil, canola oil, palm oil, olive oil and myglyol, wherein the number of fatty acid carbons is between 12 and 22 and wherein the fatty acids can be saturated or unsaturated.
- charged lipid or phospholipid can be suspended in the neutral oil.
- DNase free phosphatidylcholine is added to DNase free triglyceride soybean oil at a ratio of 1 gram of phospholipid to 20 ml of triglyceride and is dissolved by gentle heating at 50°-60°C.
- MCC phospholipid-triglyceride solution
- the suspension is incubated for 60 min. at 20°C and is then mixed with DNase-free PBS in the ratio of 20 ml MCC suspension per liter of PBS.
- the mixture is emulsified by sonication at 20% output for 5 min (Model W-385 Sonicator, Heat Systems-Ultrasonics Inc.).
- the emulsified mixture is homogenized by microfluidization at 15,000-30,000 psi for one or more flow-throughs (Model M-110Y; Microfluidics).
- the MCC emulsion is transferred to an autoclaved, capped bottle for storage at 4°C.
- a therapeutic agent or a stabilizing agent can be added to the M-DNA or the MCC composition during incubation, emulsification or homogenization.
- M-DNA and MCC are administered in an amount effective to have an anti- neoplastic effect on cancer cells in the prostate of an animal.
- the dosage of M-DNA and MCC are administered in an amount effective to have an anti- neoplastic effect on cancer cells in the prostate of an animal.
- MCC administered will depend on the stage of the prostate cancer being treated, the organs to which it may have metastasized, the particular formulation, and other clinical factors such as the size, weight and condition of the recipient and the route of administration.
- the amount of M-DNA and MCC administered is from about 0.00001 to
- the M-DNA content of MCC is between about 0.001 and 90 mg/100 mg dry MCC, more preferably between about 0.01 and
- Routes of administration include, but are not limited to, oral, topical, subcutaneous, intra-prostatic, intra-muscular, intra-peritoneal, intra-venous, intra- arterial, intra-dermal, intra-thecal, intra-lesional, intra-tumoral, intra-bladder, intra- vaginal, intra-ocular, intra-rectal, intra-pulmonary, intra-spinal, transdermal, subdermal, placement within cavities of the body, nasal inhalation, pulmonary inhalation, impression into skin and electrocorporation.
- the volume per dose is preferably about 0.001 to 100 ml per dose, more preferably about 0.01 to 50 ml per dose and most preferably about 0.1 to 30 ml per dose.
- M-DNA and MCC can be administered in a single dose treatment or in multiple dose treatments on a schedule and over a period of time appropriate to the stage of the prostate cancer being treated, the organs to which it has metastasized, the condition of the recipient and the route of administration.
- MCC was prepared from M. phlei as described in International Patent Application No. PCT/ CA98/ 00744, which is included by reference herein. All reagents were selected to preserve the DNA.
- M-DNA was prepared from M. phlei and from MCC as described in International Patent Application No. PCT/CA98/00744, which is included by reference herein. All reagents were selected to enhance conservation of (preserve) the DNA.
- M-DNA and MCC were suspended in DNase-free water or in a pharmaceutically acceptable DNase-free buffer and sonicated.
- the particle size of MCC was evaluated during sonication by photon correlation spectroscopy (Zetasizer 3000, Malvern Instruments, Malvern, Worcester, England).
- the diameter of MCC decreased gradually with sonication (Table 1).
- the molecular weight of M-DNA was evaluated during sonication by electrophoresis in 2.0 % agarose gel containing 0.5 ⁇ g/ml ethidium bromide (3 hours at 100 V).
- the molecular weight of M-DNA decreased gradually with sonication (Table 2).
- M-DNA and MCC did not contain endotoxins as determined using a Limulus amebocyte lysate QCL-1000 kit (BioWhittaker, Walkersville, MD).
- the Limulus amebocyte lysate sensitivity of this assay is 5.5 pg endotoxin/ml.
- B-DNA and BCC are prepared from M. smegmatis, M. fortuitous, Nocardia rubra, Nocardia asteroides, Cornybacterium parvum, M. kansaii, M tuberculosis and M. bovis as in Example 1.
- DNase I digests both single stranded and double stranded DNA and causes almost total degradation of the DNA.
- PC3 and DU-145 androgen-independent and LNCaP androgen-dependent prostate cancer cells were obtained from the ATCC (#CRL-1435, HTB-81 and CRL-1740, respectively).
- Primary human prostate epithelial cells (PrEc) were obtained from Clonetics (#CC-2555, Walkersville, MD, USA). All cell lines were cultured as recommended by the supplier.
- PC3, DU-145, LNCaP and PrEc cells were seeded at 3 X 10 5 cells/ml of growth medium in 6-well flat bottom microplates and allowed to grow for 24 h at 37°C in 5% C0 2 . At 24 h, growth medium was replaced with growth medium containing M-DNA or MCC.
- MTT dimethylthiazoldiphenyltetrazolium bromide
- the absorbence of the reaction product was determined at 570 nm.
- MCC at 1, 10 and 100 ⁇ g/ml inhibited proliferation of androgen-dependent LNCaP prostate cancer cells and of androgen-independent PC3 and DU-145 prostate cancer cells, but did not inhibit proliferation of PrEc normal prostate cells (Fig. 1).
- MCC inhibits proliferation of both androgen- independent and androgen-dependent prostate cancer cells in the absence of immune effector cells. MCC does not inhibit proliferation of normal prostate cells.
- M-DNA inhibits proliferation of androgen-dependent and androgen-independent prostate cancer cells in the absence of immune effector cells.
- PC3 prostate cancer cells were incubated with 1 Unit/ml DNase I, with 1 ⁇ g/ml MCC and with 1 ⁇ g/ml DNase I treated MCC.
- DNase I inhibited proliferation PC3 cancer cells about 5%
- MCC inhibited proliferation PC3 cancer cells about 50%
- DNase I treated MCC inhibited proliferation of PC3 cancer cells about 30% (Fig. 3).
- Morphological changes indicative of cell death by apoptosis include condensation of nuclear chromatin, cell shrinkage, nuclear disintegration, plasma membrane blebbing and the formation of membrane-bound bodies (Wyllie et al. Int. Rev. Cytol. 68:251,1980).
- PC3 prostate cancer cells were incubated with 0 and with 200 ⁇ g/ml of MCC for 48 h. Images were collected on a light microscope with a 40 x 2.5 NA Apochromat objective.
- the pellet of detached cells and the remaining attached cells were lysed in 50 ⁇ l of buffer containing 50 mM Tris-HCl (pH 8.0), 10 mM EDTA, 0.5 % (w/v) sodium lauryl sarcosinate (SDS) and 0.5 mg/ml proteinase K (Sigma-Aldrich Canada, Oakville, Ontario) at 55°C for 1 h.
- Samples were heated to 65°C and 10 ⁇ l of 10 mM EDTA (pH 8.0) containing 1 % (w/v) low-gelling-temperature agarose, 0.25 % (w/v) bromophenol blue, 40 % (w/v) sucrose was mixed with each sample.
- the samples were electrophoresed at 100 V for 3 h in a 2 % agarose gel containing Tris-borate/EDTA buffer (TBE).
- TBE Tris-borate/EDTA buffer
- Detached and attached PC3 prostate cancer cells were incubated for 48 h with 0 and 100 ⁇ g/ml MCC. Detached cells treated with 100 ⁇ g/ml MCC showed significant
- the induction of apoptosis may also be demonstrated by the solubilization and release of nuclear matrix (NuMA) protein.
- LNCaP prostate cancer cells were incubated for 48 h with 0, 100, 200 and 300 ⁇ g/ml MCC while PC3 prostate cancer cells were incubated for 48 h with 0, 100 and 300 ⁇ g/ml MCC.
- Medium was drawn off, centrifuged at 10,000 X G and the supernatant was frozen at -20°C until assayed for NuMA (Miller et al. Biotech. 15:1042, 1993).
- LNCaP prostate cancer cells treated with 100, 200, or 300 ⁇ g/ml MCC released 20%, 50% and 135%, respectively, more NuMA proteins into the medium than did cells treated with 0 ⁇ g/ml MCC (Fig. 6A).
- PC3 prostate cancer cells treated with 100 and 300 ⁇ g/ml MCC released 24% more NuMA proteins into the medium than did cells treated with 0 ⁇ g/ml MCC (Fig. 6B).
- THP-1 cells Human monocytic THP-1 cells were obtained from the ATCC and were cultured as recommended by the supplier. THP-1 cells (1 x 10 6 cells/ml) and LNCaP cells (3 x 10 5 cells/ml) were incubated with 0, 0.1 and 10 ⁇ g/ml MCC for 48 hours in 5% C0 2 in 6-well flat bottom microplates. At 48 hours, supernatants were collected and the presence of IL- 12 and TNF- ⁇ was determined using commercial ELISA kits (Biosource, Camarillo, CA, USA). MCC and M-DNA induced synthesis of IL-12 (Table 3) and TNF- ⁇ Table 4 by immune system cells, THP-1 monocytes, and by LNCaP prostate tumor cells.
- cytokines IL-12 and TNF- ⁇ both have been demonstrated to have anti-neoplastic activity against a range of cancer cells, including prostate cancer cells (Izquierdo et al. Anticancer Drugs 7: 275, 1996; Sensibar et al. Cancer Res. 55: 2431, 1995).
- EXAMPLE 12 Effects of MCC, M-DNA and DNase I treated MCC on PC3 tumors in mice Androgen-independent PC3 prostate cancer cells are implanted subcutaneously into 40 male nude BALB/c mice and allowed to grow until palpable (0.1 to 0.5 cm in diameter). The mice are divided into 4 groups and tumor mass is measured in each mouse.
- the 10 mice in Group 1 each receive saline.
- the 10 mice in Group 2 each receive saline containing MCC.
- the 10 mice in Group 3 each receive saline containing M-DNA.
- the 10 mice in Group 4 each receive DNase I treated MCC. After 4 weeks of once per week intra- tumoral treatments, the mice are sacrificed and the tumor mass and number of metastases are determined. The mice in Group 2 and in Group 3 have less tumor mass and fewer metastases than the mice in Group 1 and in Group 4.
- EXAMPLE 13 Effects of MCC and of M-DNA alone and in combination with estramustine and etoposide on PC3 tumors in mice Androgen-independent PC3 prostate cancer cells are implanted subcutaneously into 60 male nude BALB/c mice and allowed to grow until palpable (0.1 to 0.5 cm in diameter). The mice are divided into 6 groups and tumor mass is measured in each mouse.
- the 10 mice in Group 1 receive saline.
- the 10 mice in Group 2 receive MCC in saline.
- the 10 mice in Group 3 receive M-DNA in saline.
- the 10 mice in Group 4 receive estramustine and etoposide in saline.
- the 10 mice in Group 5 receive MCC in combination with estramustine and etoposide in saline.
- mice in Group 6 receive M-DNA in combination with estramustine and etoposide in saline. After 4 weeks of once per week intra-tumoral treatment the mice are sacrificed and tumor mass and number of metastases are determined.
- the mice in Group 1 have the most tumor mass.
- the mice in Group 4 have less tumor mass than the mice in Group 1.
- the mice in Group 2 and Group 3 have less tumor mass than the mice in Group 4.
- the mice in Group 5 and in Group 6 have the least tumor mass.
- Androgen-dependent LNCaP prostate cancer cells are implanted subcutaneously into 40 male nude BALB/c mice and allowed to grow until palpable (0.1 to 0.5 cm in diameter).
- the mice are divided into 4 groups and tumor mass is measured in each mouse.
- the 10 mice in Group 1 each receive saline.
- the 10 mice in Group 2 each receive saline containing MCC.
- the 10 mice in Group 3 each receive saline containing M-DNA.
- the 10 mice in Group 4 each receive DNase I treated MCC. After 4 weeks of once per week intra- tumoral treatment the mice are sacrificed and the tumor mass and number of metastases are determined.
- the mice in Group 2 and in Group 3 have less tumor mass and fewer metastases than the mice in Group 1 and in Group 4.
- EXAMPLE 15 EXAMPLE 15
- Androgen-dependent LNCaP prostate cancer cells are implanted subcutaneously into 60 male nude BALB/c mice and allowed to grow until palpable (0.1 to 0.5 cm in diameter).
- the mice are divided into 6 groups and tumor mass is measured in each mouse.
- the 10 mice in Group 1 receive saline.
- the 10 mice in Group 2 receive MCC in saline.
- the 10 mice in Group 3 receive M-DNA in saline.
- the 10 mice in Group 4 receive flutamide in saline.
- the 10 mice in Group 5 receive MCC in combination with flutamide in saline.
- the 10 mice in Group 6 receive M-DNA in combination with flutamide in saline.
- mice After 4 weeks of once per week intra-tumoral treatment the mice are sacrificed and the tumor mass and number of metastases are determined.
- the mice in Group 1 have the most tumor mass.
- the mice in Group 4 have less tumor mass than the mice in Group 1.
- the mice in Group 2 and Group 3 have less tumor mass than the mice in Group 4.
- the mice in Group 5 and in Group 6 have the least tumor mass.
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Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
AT00912310T ATE296106T1 (en) | 1999-04-01 | 2000-03-30 | COMPOSITION AND METHOD FOR INDUCING APOPTOSIS IN PROSTATE CANCER CELLS USING M-DNA AND MCC |
US09/937,643 US6794368B1 (en) | 1999-04-01 | 2000-03-30 | Composition and method for inducing apoptosis in prostate cancer cells |
CA2366090A CA2366090C (en) | 1999-04-01 | 2000-03-30 | Composition and method for inducing apoptosis in prostate cancer cells |
DE60020352T DE60020352T2 (en) | 1999-04-01 | 2000-03-30 | Composition and method of inducing apptosis in prostate cancer cells using M-DNA and MCC |
DK00912310T DK1165106T3 (en) | 1999-04-01 | 2000-03-30 | Composition and Method of Inducing Apoptosis in Prostate Cancer Cells by M-DNA and MCC |
AU34135/00A AU780909B2 (en) | 1999-04-01 | 2000-03-30 | Composition and method for inducing apoptosis in prostate cancer cells |
JP2000609081A JP2002541114A (en) | 1999-04-01 | 2000-03-30 | Compositions and methods for inducing apoptosis in cells of prostate cancer |
EP00912310A EP1165106B8 (en) | 1999-04-01 | 2000-03-30 | Composition and method for inducing apoptosis in prostate cancer cells by m-dna and mcc |
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US12745199P | 1999-04-01 | 1999-04-01 | |
US60/127,451 | 1999-04-01 |
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WO2000059518A2 true WO2000059518A2 (en) | 2000-10-12 |
WO2000059518A3 WO2000059518A3 (en) | 2001-02-01 |
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PCT/CA2000/000342 WO2000059518A2 (en) | 1999-04-01 | 2000-03-30 | Composition and method for inducing apoptosis in prostate cancer cells |
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EP (1) | EP1165106B8 (en) |
JP (1) | JP2002541114A (en) |
AT (1) | ATE296106T1 (en) |
AU (1) | AU780909B2 (en) |
CA (1) | CA2366090C (en) |
CY (1) | CY1107455T1 (en) |
DE (1) | DE60020352T2 (en) |
ES (1) | ES2238996T3 (en) |
PT (1) | PT1165106E (en) |
WO (1) | WO2000059518A2 (en) |
Cited By (1)
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WO2002085410A2 (en) * | 2001-04-24 | 2002-10-31 | Bioniche Life Sciences, Inc. | Method for evaluating the efficacy of treatment with bacterial dna and bacterial cell walls |
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JP4488626B2 (en) * | 1998-12-04 | 2010-06-23 | バイオニケ ライフ サイエンシーズ インコーポレイテッド | Inflammation treatment |
Citations (1)
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WO1999007383A1 (en) * | 1997-08-05 | 1999-02-18 | Bioniche Inc. | Composition and method for regulating cell proliferation and cell death |
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- 2000-03-30 ES ES00912310T patent/ES2238996T3/en not_active Expired - Lifetime
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- 2000-03-30 AU AU34135/00A patent/AU780909B2/en not_active Ceased
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WO1999007383A1 (en) * | 1997-08-05 | 1999-02-18 | Bioniche Inc. | Composition and method for regulating cell proliferation and cell death |
Non-Patent Citations (5)
Title |
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FILION M C ET AL: "MYCOBACTERIAL CELL WALL - DNA COMPLEX INDUCES APOPTOSIS IN CANCER CELLS" JOURNAL OF PHARMACY AND PHARMACOLOGY,LONDON,GB, vol. 50, no. SUPPL, September 1998 (1998-09), page 39 XP000892394 ISSN: 0022-3573 * |
FILION M C ET AL: "Mycobacterium phlei cell wall complex directly induces apoptosi in human bladder cancer cells" BRITISH JOURNAL OF CANCER,GB,LONDON, vol. 79, no. 2, January 1999 (1999-01), pages 229-235, XP002100762 ISSN: 0007-0920 * |
FILION M C ET AL: "Mycobacterium phlei cell wall complex, a new anti-tumoral agent, induces IL-12 synthesis by monocyte-macrophages via associated DNA" BLOOD,US,W.B. SAUNDERS, PHILADELPHIA, VA, vol. 90, no. 10, SUPPL. 01, 5 December 1997 (1997-12-05), page 58B XP002085542 ISSN: 0006-4971 * |
MORALES A ET AL: "Immunotherapy of an experimental adenocarcinoma of the prostate" JOURNAL OF UROLOGY,US,BALTIMORE, MD, vol. 153, May 1995 (1995-05), pages 1706-1710, XP002100764 ISSN: 0022-5347 * |
READER S ET AL: "Mycobacterial cell wall-DNA complex (MCC) inhibits proliferation and induces apoptosis in androgen-dependent and independent human prostate cancer cells." BRITISH JOURNAL OF CANCER, vol. 80, no. SUPPL. 2, July 1999 (1999-07), page 76 XP000946798 Joint Meeting of the British Association for Cancer Research, the British Oncological Association, the Association of Cancer Physicians and the Royal College of Radiologists;Edinburgh, Scotland, UK; July 11-14, 1999 ISSN: 0007-0920 * |
Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2002085410A2 (en) * | 2001-04-24 | 2002-10-31 | Bioniche Life Sciences, Inc. | Method for evaluating the efficacy of treatment with bacterial dna and bacterial cell walls |
WO2002085410A3 (en) * | 2001-04-24 | 2003-09-18 | Bioniche Life Sciences Inc | Method for evaluating the efficacy of treatment with bacterial dna and bacterial cell walls |
Also Published As
Publication number | Publication date |
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DE60020352T2 (en) | 2005-10-20 |
CA2366090A1 (en) | 2000-10-12 |
EP1165106B8 (en) | 2005-07-27 |
EP1165106B1 (en) | 2005-05-25 |
EP1165106A2 (en) | 2002-01-02 |
CY1107455T1 (en) | 2012-12-19 |
CA2366090C (en) | 2011-05-24 |
DE60020352D1 (en) | 2005-06-30 |
WO2000059518A3 (en) | 2001-02-01 |
JP2002541114A (en) | 2002-12-03 |
ES2238996T3 (en) | 2005-09-16 |
AU3413500A (en) | 2000-10-23 |
AU780909B2 (en) | 2005-04-21 |
PT1165106E (en) | 2005-10-31 |
ATE296106T1 (en) | 2005-06-15 |
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