Classifications

• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K31/00—Medicinal preparations containing organic active ingredients
  • A61K31/33—Heterocyclic compounds
  • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
  • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
  • A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
• • 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
  • A61K31/13—Amines
  • A61K31/135—Amines having aromatic rings, e.g. ketamine, nortriptyline
• • 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
  • A61K33/00—Medicinal preparations containing inorganic active ingredients
  • A61K33/24—Heavy metals; Compounds thereof
  • A61K33/243—Platinum; Compounds 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
  • A61K38/14—Peptides containing saccharide radicals; Derivatives thereof, e.g. bleomycin, phleomycin, muramylpeptides or vancomycin
• • 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

• This invention relates to cytoprotective agents.
• it relates to the use of monoamine oxidase (MAO) inhibitors in the protection or rescue of normal cells from the toxic side effects caused by irradiation or the administration of chemotherapeutic 10 agents.
• MAO monoamine oxidase
• the MAO inhibitors can selectively protect or rescue normal cells from the toxic side effects caused by irradiation or the administration of chemotherapeutic agents. Furthermore, the MAO inhibitors may be effective at dose levels much lower than those at
• neurodegenerative conditions such as, for example, Parkinson's disease.
• a monoamine oxidase inhibitor in the preparation of a medicament for protecting and/or rescuing normal cells from toxicity resulting from irradiation and/or the administration of a chemotherapeutic agent, in particular a chemotherapeutic agent which can disrupt DNA or RNA replication.
• the invention also provides a pharmaceutical product for protecting and/or rescuing normal cells from toxicity resulting from the administration of a chemotherapeutic agent, the product comprising: (a) a monoamine oxidase inhibitor; and
• Preferred MAO inhibitors for use in the present invention are deprenyl (N-1-phenylisopropyl-N-methyl- 2-propynylamine) and structural analogues and derivatives thereof.
• Preferred MAO inhibitors are selected from the following:
• Z_ and Z 2 are each independently optionally substituted phenyl; and n is an integer from 2 to 4; and pharmaceutically acceptable salts thereof;
• Z3 is indolyl substituted at the 1- position by hydrogen, methyl or ethyl, and at the 5- position by hydrogen, hydroxy or methoxy; and X, which may be present at the 2- or 3- position of the indolyl ring, is C n H2 n -N(R.3) -R4, wherein R3 is a substituted ethylenic or acetylenic group; R4 is hydrogen, methyl or ethyl and n is an integer from 1 to 4; and pharmaceutically acceptable salts thereof;
• R is linear or branched aliphatic hydrocarbon; and pharmaceutically acceptable salts thereof ;
• Y is optionally substituted indanoyl or indolyl;
• R if present, is hydrogen or methyl;
• p is 0 or 1 ;
• n is an integer from 1 to 4 ; and pharmaceutically acceptable salts thereof;
• C ⁇ __4 alkyl refers to a saturated hydrocarbyl radical having a straight or branched chain of from 1 to 4 carbon atoms. Included within the scope of this term are methyl, ethyl, n- propyl, isopropyl, n-butyl, isobutyl and tertiary butyl .
• C1-4 alkoxy , "C 1 _ 4 thioalkyl”, “C j ⁇ alkylsulphinyl” and “C ⁇ __4 alkylsulphonyl” refer to the relevant radical, the C ⁇ _4 alkyl moiety of which is as defined above.
• the term "patient” refers to warmblooded animals or mammals including humans, who are in need of treatment to protect or rescue normal cells from the toxic side effects caused by irradiation or the administration of a chemotherapeutic agent.
• pharmaceutically acceptable acid addition salt encompasses both organic and inorganic acid addition salts including, for example, those prepared from acids such as hydrochloric, hydrobromic, sulphuric, sulphonic, tartaric, fumaric, citric, lactic, maleic, phosphoric, succinic, acetic, benzoic, ascorbic, p-toluenesulphonic, benzenesulphonic, propionic and the like.
• the hydrochloric acid addition salts are preferred.
• Particularly preferred compounds for use in the present invention are ( -) -N-1-phenylisopropyl-N- methyl-2-propynylamine (known generically as selegiline or I-deprenyl) and (-)-N-2,4- dichlorophenoxypropyl-N-methyl -2 -propynylamine (known generically as clorgyline) .
• the hydrochloric acid addition salts of these compounds are particularly preferred for use in the present invention.
• the MAO inhibitors used in the present invention can be prepared as described in Fowler, J.S., J.Org.Chem. 42, 2637 (1977); Cruces, M.A. , Eur. J. Med. Chem., 25:257 (1990); O'Brien, E.M. et al . , J. Neural Transm. [suppl.] 41, 295-305 (1994); and Yu, P.H. et al . , Adv. Exp. Med. Biol. 363: 17 (1994).
• the MAO inhibitors used in the present invention act to protect and/or rescue normal or uninfected cells from the toxic side effects caused by the administration of chemotherapeutic agents, but have no effect on tumourigenic or infected cells.
• Certain chemotherapeutic agents can be useful in the treatment of diseases. However, these agents can also cause toxic side effects that can limit their use and/or effectiveness. Examples of such agents are DNA- intercalating agents (e.g. cisplatin) , pyrimidine bases (e.g. 5-fluorouracil) or DNA strand scission inducing agents (e.g. bleomycins) used in the treatment of cancers; and pyrimidine nucleosides (e.g. azidodeoxythymidine (AZT) ) used in the treatment of AIDS.
• DNA- intercalating agents e.g. cisplatin
• pyrimidine bases e.g. 5-fluorouracil
• DNA strand scission inducing agents e.g
• the MAO inhibitors may be administered simultaneously, separately or sequentially with chemotherapeutic agents of the above type.
• the MAO inhibitors are conveniently administered prior to the administration of the chemotherapeutic agent, for example, at least three hours prior thereto.
• the MAO inhibitors also protect and/or rescue normal cells from toxicity resulting from irradiation, such as gamma-radiation, including radiation therapy used in the treatment of certain cancers.
• the MAO inhibitors are conveniently administered prior to irradiation, for example, at least three hours prior thereto, especially in the treatment of cancer.
• MAO inhibitors to protect/rescue normal cells from the toxic side effects of irradiation or the administration of chemotherapeutic agents allows a more cytotoxic dose of radiation or chemotherapeutic agent to be administered while the surrounding normal cells remain biologically intact.
• MAO inhibitors are ideal agents for use in therapy requiring the use of radiation or chemotherapeutic agents, especially in the treatment of cancer.
• the MAO inhibitors may be administered orally, for example with an inert diluent or with an assimilable edible carrier. They may also be enclosed in hard or soft shell gelatin capsules or they may be compressed into tablets.
• the MAO inhibitors may be incorporated with pharmaceutically acceptable excipients and used in the form of ingestible tablets, buccal tablets, capsules, suspensions, syrups and the like. Suitable excipients include solvents such as water, alcohol and propylene glycol, surface active agents, suspending agents, lubricants, flavouring agents, colourants, and the like. Such carriers and excipients are well known to those skilled in the art.
• the MAO inhibitors may also be administered parenterally (e.g. intravenously).
• injectable dosage forms of a solution or suspension of the MAO inhibitors can be prepared for example in a physiologically acceptable diluent with a pharmaceutical carrier which can be a sterile liquid such as water and oils, with or without the addition of a surfactant and other pharmaceutically acceptable adjuvants.
• a pharmaceutical carrier which can be a sterile liquid such as water and oils, with or without the addition of a surfactant and other pharmaceutically acceptable adjuvants.
• oils which can be employed in these preparations are those of petroleum, animal, vegetable or synthetic origin, for example, peanut oil, soybean oil and mineral oil.
• water, saline, aqueous dextrose and related sugar solutions, ethanols and glycols are preferred liquid carriers, particularly for injectable solutions.
• an oral dosage could be as low as 70 ng (or even less) to 700 ⁇ g, preferably from 1 ⁇ g (or less) to 100 ⁇ g, per day for an average adult patient (70 kg) .
• Single dosages for parenteral (e.g. intravenous) administration could be as low as 10 ng (or even less) to 100 ⁇ g, preferably from 100 ⁇ g (or less) to 50 ⁇ g, for a 70 kg patient.
• the compound (-)-N-l- phenylisopropyl-N-methyl-2 -propynylamine hydrochloride is referred to as deprenyl
• the compound (-)-N- 2,4-dichlorophenoxypropyl-N-methyl-2 -propynylamine hydrochloride is referred to as clorgyline.
• deprenyl and clorgyline are added to cell line cultures immediately after plating and to explant cultures immediately after attachment .
• Collagenase type IV Collagenase type IV; clorgyline; 5-fluorouracil; hydrocortisone; insulin; hydrogen peroxide; and diaminobenzidene (DAB) were obtained from Sigma Chemical Company.
• FCS Foetal calf serum
• HS Horse serum
• HEPES Buffer Versene (Trademark; 0.2g/l EDTA in isotonically buffered saline); Hank's balanced salt solution (0.4 g/1 KCl, 0.06 g/1 KH 2 P0 , 8.0 g/1 NaCl ,
• Glycergel was prepared by the inventors by dissolving 10 g gelatine (BDH) in 60 ml distilled water and adding 70 ml glycerol and 0.25 g phenol. The resulting product becomes solid and requires melting before use.
• Isoton (Trademark) is a zwitterionic solution supplied by the Coulter Electronic Company and contains 7.9 g/1 NaCl, 1.9 g/1 Na 2 HP0 4 , 0.4 g/1 EDTA disodium salt, 0.4 g/1 KCl.
• Deprenyl was obtained from Semat Technical, St. Albans, England.
• Cisplatin was obtained from David Bull Laboratories.
• Bleomycin sulphate was obtained from Lundbeck, United Kingdom.
• 5-Fluorouracil was obtained from Roche.
• PBS Phosphate buffered saline
• the HACAT cell line was derived from human skin keratinocytes spontaneously transformed in vi tro during long term incubation of a primary culture under selected culture conditions. This cell line was obtained as a gift from Dr. P. Boukamp (Heidelberg University, Germany) .
• the HPV-G (human papilloma virus-G) cell line was obtained as a gift from Dr. J. Dipaolo, NIH, (Bethesda, MD, USA) . This line was transformed from human neonatal foreskin transfected with the HPV-16 virus to cause immortalisation.
• the HPV-G cell line is also available from the European Collection of Cell Cultures (ECACC) .
• the CHO-K1 cell line was established from a subclone of the parent CHO cell line which was derived from the ovary of a Chinese hamster and was supplied by the ECACC.
• the PC3 cell line was established from a grade 4 prostatic adenocarcinoma from a sixty-two year old male Caucasian and this cell line was obtained from the ECACC.
• the stock cells were grown in 260 ml nunc flasks in the appropriate growth medium for the particular cell line involved. To avoid rapid loss of differentiation, the cells were not passaged too often or split at too high a ratio. When the cells reached confluence, they were passaged in the following way: cells were rinsed for approximately one minute with 0.25% w/v trypsin solution (2.5% stock trypsin diluted 1:10 with Hank's balanced salt solution) and 1 mM Versene solution in a 1:1 mixture. This solution was then poured off and a fresh 0.25% w/v trypsin/lmM Versene solution (1:1) was used to trypsinise the cells at 37°C for approximately 12-15 minutes depending on the cell line.
• the plating efficiency is the fraction of cells plated that grow to form colonies after significant time has elapsed i.e. "stainable groups of cells, surface attached, and large enough to be recognised by the eye" [Elkind, M.M., Whitmore, G.S., The Radiobiol ogy of Cultured Mammalian Cells (1967, New York, Gordon & Breach) ] .
• the plating efficiency was also determined per dose point at 18-20 days (i.e. the time taken for at least 15 cell doublings) post irradiation. Appropriate cell numbers were seeded into six flasks per dose point. These cells were allowed to attach to the surface of the flask and were then irradiated after 6 hours. When the cells produced macroscopic colonies (9-10 days) , three of the original six flasks were stained with carbol- fuschin, and the colonies were counted. The medium was then removed from the remaining three flasks and a fresh 5 ml of culture medium was placed into each flask.
• % Residual surviving fraction Initial % S.F. x Pm ⁇ pn ⁇ % S.F.
• the residual percentage surviving fraction is a measure of the level of survival at times distant, i.e. 18-20 days, from the initially irradiated cells.
• All connective tissue was removed from the urether or bladder biopsy using a scalpel. The tissue was then cut into 2-3 mm 2 pieces using a scalpel. This tissue was then placed in 50 mg collagenase type IV/5 ml 0.25% w/v trypsin solution (2.5% stock trypsin diluted 1:10 with Hank's balanced salt solution) for 30 minutes at 37°C. The digested explants were then placed singly in 25 cm 2 nunc flasks with 2 ml of growth medium. When attachment occurred after approximately 3 days, the medium was removed and replaced with serum free medium to inhibit the growth of fibroblasts. Deprenyl or clorgyline was also added at this stage. Cultures were grown for 14 days before irradiation and fixation for immunocytochemical analysis .
• Immunostaining was performed using a Vectastain ABC kit which employs a standard immunoperoxidase method.
• the cell culture was washed in phosphate buffered saline (PBS) and the endogenous peroxidase activity blocked with 3% H2O2 in PBS for 5 minutes.
• the culture was then washed in buffer for 20 minutes. All excess serum was washed from the sections and the slides were incubated with primary antibody diluted (1:20) in PBS for 60 minutes.
• the slides were washed again in buffer for 10 minutes.
• the sections were incubated for 30 minutes with diluted biotinylated antibody solution from the Vectastain ABC kit and then washed for 10 minutes in PBS buffer.
• Sections were then incubated for 45 minutes with Vectastain ABC reagent and washed in PBS buffer for a further 10 minutes. Sections were incubated in diaminobenzidine (DAB) prepared as follows : -1000 ⁇ l of DAB, 900 ⁇ l of PBS and 10 ⁇ l of H 2 0 2 until staining became apparent. The slides were then washed in tap water and counter- stained with haematoxylin for 1 minute. The sections were then washed in hot water and mounted in glycergel. A positive reaction was indicated by a brown staining.
• DAB diaminobenzidine
• Urether and Bladder Explant culture medium RPMI-1640 plus 7.5% FCS and 10% HS 5 ml L-Glutamine (200mM)
• the monoamine oxidase (MAO) inhibitors deprenyl and clorgyline, were made up in distilled water to the required molarities and were added to the cell - culture flasks at least 3 hours before irradiation to allow these MAO inhibitors to take effect prior to irradiation.
• the standard volume of inhibitor which was added to the required flask was always 100 ⁇ l .
• 100 ⁇ l of distilled water were added to keep the volume of cell culture medium constant throughout the experiment .
• the same protocol was employed when conducting the chemotherapy studies and the hydrogen peroxide induced cell death studies.
• the chemotherapeutic agent, cisplatin consisted of a 1.0 mg/ml solution of cisplatin made up in a sodium chloride/ mannitol solution. In order to achieve the correct concentration of cisplatin, this stock solution was diluted with distilled water. 5- Fluorouracil was supplied in a solid form and was dissolved and made up to the desired concentration with distilled water. Bleomycin sulphate was dissolved in distilled water to give a 20 mg/ml stock solution which was diluted in medium as required. All additions of chemotherapeutic agents were made 6 hours after the cells had attached to the flasks and were incubated with the cells for 1 hour.
• the- medium was poured off and the cells in monolayer culture were washed with Hank's balanced salt solution for 1 minute approximately. This solution was then poured off and the flask was re-filled with 5 ml of the appropriate cell culture medium.
• Hydrogen peroxide was supplied in a 30% w/w solution.
• the desired concentration of hydrogen peroxide was obtained by diluting appropriately with distilled water. All additions of hydrogen peroxide to cell culture flasks were made 6 hours after the cells were plated initially.
• Human skin keratino ⁇ ytes The cell lines used were HACAT cells which are a non-tumourigenic but immortalized cell line and human papilloma virus (HPV) transfected keratinocytes which are tumourigenic.
• HPV human papilloma virus
• Deprenyl and clorgyline were added immediately after plating and the cells were irradiated 6 hours after plating at the dosage indicated with cobalt-60 gamma radiation.
• the results in Tables 2a, 2b and 2c show that both deprenyl and clorgyline give rise to radio- protection and enhanced cloning efficiency in the normal, but not the tumourigenic, cell line.
• the results in Table 2d show that significant protection is obtained in HaCaT cells at 10 " and 10 " M deprenyl over a range of radiation doses from 2.5 Gy (therapeutic) to 10 Gy.
• Apoptosis gene expression Cultures exposed to deprenyl and clorgyline were fixed and stained 1 hour after irradiation for expression of the product of the gene bcl-2 involved in damage control and repair.
• the results in Tables 6a- 6f show very strong induction of bcl-2 in the deprenyl and clorgyline treated cultures whether exposed to radiation or not. Controls induced bcl-2 expression following irradiation but this induction took at least one hour.
• HPV-G cells Human keratinocyte cell line, HPV 16 transfected, immortalised, tumourigenic irradiated at 6 hours post plating.
• HPV-G cells Human keratinocyte cell line, HPV 16 transfected, immortalised, tumourigenic .
• the cells were irradiated under the same conditions as the experiments in Table 2b but were then allowed to go through at least 15 cell doublings prior to assay.
• PC3 cells human prostate carcinoma cells
• Clonogenic assay results for HaCaT cell line human keratinocyte non-tumourigenic cell line
• cisplatin DNA intercalating agent used in cancer therapy
• Clonogenic assay results for CHO-Kl cell line Choinese Hamster Ovary tumourigenic cell line
• 4 6, 8 and 10 microgrammes per ml of cisplatin (DNA intercalating agent used in cancer therapy) for 1 hour 6 hours post plating and treated with Deprenyl at a concentration of 1 nanomolar where indicated immediately after plating.
• HaCaT cells normal human keratinocytes
• PC3 cells human prostate carcinoma
• HPV-G cells Human tumourigenic keratinocytes
• CHO-Kl cells Choinese Hamster ovary established cell line, immortalised, tumourigenic
• Clonogenic assay results for HaCaT cell line (human keratinocyte non- tumourigenic cell line) treated with 100 to 400 micromolar solutions of hydrogen peroxide to promote oxidative stress induced cell death 6 hours after plating and treated with Deprenyl at a concentration of 1 nanomolar where indicated immediately after the cells were plated.
• the MAO inhibitors also have a protective effect on non-tumourigenic cells treated with chemotherapeutic agents, but have no such protective effect on tumourigenic cells treated with said agents. Thus, a more cytotoxic dose of radiation or chemotherapy can be delivered to a tumour while the surrounding normal cells remain protected.
• the MAO inhibitors can also rescue damaged cells making them fit to deal with damage rather than improve survival of already fit cells. This is dramatically illustrated by the behaviour of the HACAT cell line plated at low cell numbers. As shown in Table 2a, the plating efficiency of this cell line is always relatively low and under these conditions plating efficiencies in the range ⁇ 0.002 - 0.08% were obtained in the absence of deprenyl or clorgyline. Plating under exactly the same conditions but in the presence of 10 ⁇ 9 M deprenyl or clorgyline had the effect that plating efficiencies increased to approximately 1.9 and 4.0% respectively. These survival figures were not greatly affected by radiation at either 0.5 or 5.0 Gy (plating efficiencies 2.6 and 2.2, respectively, in the presence of 10 ⁇ 9 M deprenyl) .
• the mechanism of action of the MAO inhibitors is at present unknown.
• Table 6c showing very strong induction of the bcl-2 gene after irradiation of normal cells implicate the activation of anti-apoptosis pathways.
• Tumour cells generally have bcl-2 already induced thus increasing their resistance to damage.
• one action of the MAO inhibitors could be to induce this anti-apoptotic pathway in the, relatively radiosensitive, normal cells, thereby increasing their survival to a level comparable to the more radio-resistant tumour cells.
• Another theory is that the MAO inhibitors could down regulate the expression of another protein known as BAX, in normal cells.
• BAX and bcl-2 act as a rheostat in the apoptotic process since when bcl-2 is in excess within the cell, apoptosis is inhibited. However, when BAX is in excess apoptosis proceeds. These two proteins cooperate and bind to each other via their BH1 and BH2 protein domains and the effect that BAX has on the apoptotic process is to act somewhat like a mopping device for bcl-2, to drive the cell in the direction of apoptosis since when in complex with bcl-2 it renders this protein inactive, thereby preventing it from exerting its apoptotic repressive effects. If the MAO inhibitors down regulated BAX expression in normal cells, more bcl-2 would be available and hence one would see a higher degree of apoptotic inhibition. In effect, this is what is seen from the present results.
• HIV acquired immunodeficiency syndrome
• HIV primarily infects CD4 expressing T cells, including helper T cells and macrophages .
• the massive immunosu- pression is a result of virus induced apoptosis of subsets of T cells.
• Certain autoimmune diseases are also largely dependent on the apopototic response.
• Cytotoxic T lymphocytes which do not recognise other cells within the body as "self” excrete a toxin which may be lymphotoxin or a related molecule.
• the toxin kills cells by activating target cell enzymes that cleave DNA in the target cell nucleus. Once the nuclear DNA is fragmented, target cell nuclei also undergo fragmentation, i.e. apoptosis.
• Apoptosis is also a major cause of cell loss in biotechnological processes involving mass cell culture. It is a major cause of cell death in critical tissues such as blood, gut and liver following exposure to certain classes of toxic pollutants, e.g. cadmium or nickel, and also as a result of ischaemia.
• toxic pollutants e.g. cadmium or nickel
• the anti-apoptosis function of MAO inhibitors makes them of potential use in all of these areas .

Landscapes

• Health & Medical Sciences (AREA)
• Life Sciences & Earth Sciences (AREA)
• General Health & Medical Sciences (AREA)
• Veterinary Medicine (AREA)
• Pharmacology & Pharmacy (AREA)
• Epidemiology (AREA)
• Medicinal Chemistry (AREA)
• Animal Behavior & Ethology (AREA)
• Chemical & Material Sciences (AREA)
• Public Health (AREA)
• Inorganic Chemistry (AREA)
• Molecular Biology (AREA)
• Gastroenterology & Hepatology (AREA)
• Engineering & Computer Science (AREA)
• Bioinformatics & Cheminformatics (AREA)
• Immunology (AREA)
• Proteomics, Peptides & Aminoacids (AREA)
• Acyclic And Carbocyclic Compounds In Medicinal Compositions (AREA)

Applications Claiming Priority (3)

Publications (1)

Family

ID=11041418

Family Applications (1)

Country Status (4)

Families Citing this family (17)

Family Cites Families (2)

• 1998
  • 1998-03-13 AU AU64171/98A patent/AU6417198A/en not_active Abandoned
  • 1998-03-13 EP EP98909714A patent/EP0975366A1/de not_active Withdrawn
  • 1998-03-13 WO PCT/IE1998/000023 patent/WO1998040102A1/en not_active Application Discontinuation
  • 1998-03-13 CA CA002284001A patent/CA2284001A1/en not_active Abandoned

Non-Patent Citations (1)

Also Published As

Similar Documents

Legal Events