NZ503921A

NZ503921A - Use of fluconazole mixed with chemotherapeutic agents for inhibiting the growth of cancers or tumors

Info

Publication number: NZ503921A
Application number: NZ503921A
Authority: NZ
Inventors: James Berger Camden
Original assignee: Procter & Gamble
Priority date: 1995-08-04
Filing date: 1996-07-30
Publication date: 2002-03-01
Also published as: AU6683396A; TR199801739T2; HUP9903420A2; PL324904A1; HUP9903420A3; EP0841921A2; AR003175A1; JPH11510187A; BR9609966A; NO980473L; WO1997005873A3; CN1195288A; IL123095A0; NO980473D0; CZ33798A3; MX9800998A; AU711966B2; CA2229024A1; SK14198A3; KR19990036138A

Abstract

The use of a compound selected from the group consisting of2-(2,4-difluorophenyl)-1,3-bis(1H-1,2,4-triazol-1-yl)propan-2-ol, a derivative thereof, and mixtures thereof; and a chemotherapeutic agent, in the manufacture of a pharmaceutical composition for the treatment of cancer or tumours.

Description

<div class="application article clearfix" id="description"> New Zealand Paient Spedficaiion for Paient Number 503921 intellectual property office of n.z. 3 0 JUL 2001 RECEIVED NEW ZEALAND PATENTS ACT, 1953 No: Divided out of No. 315184 Date: Dated 30 July 1996 COMPLETE SPECIFICATION USE OF FLUCONAZOLE FOR INHIBITING THE GROWTH OF CANCERS OR TUMORS We, THE PROCTER & GAMBLE COMPANY, a corporation organised and existing under the laws of the State of Ohio, United States of America, having a place of business at One Procter & Gamble Plaza, Cincinnati, Ohio 45202, United States of America do hereby declare the invention for which we pray that a patent may be granted to us, and the method by which it is to be performed, to be particularly described in and by the following statement: - 1 - (followed by 1 A) WO 9- 058-3 10 5Q 3 S2.1 PCT'"LS96/I24?4 1 A INTELLECTUAL PROPERTY OFFICE OF NZ. 3 0 JIJ[ 2001 RECEIVED technical fteld This invention is a pharmaceutical composition that is useful for the treatment of cancers or tumors, particularly in human and warm blooded animals containing 2-{2,-i-dLfluorophenyl)-l,3-bis( 1H- 1,2,4-tnazol-1 -yi)propan-2-oi and its derivatives. It can be used in combination with other chemothexapeutic agents and potentiators. The same composition can be "<cd to treat viral infections.* The claims of this specification relate to the treatment of cancers or tumors. BACKGROUND OF THE INVENTION Cancers, including leukemia, are the leading cause of death in animals and humans. The exact cause of leukemia is not known, but links between certain activities such as smoking or 15 exposure to carcinogens and the incidence of certain types of leukemia and tumors has been shown by a number of researchers. Many types of chemotherapeuuc agents have been shown to be effective against cancers, tumors and leukemia, but not all types of cancer and tumor cells respond to these agents. Unfortunately, many of these agents also destroy normal cells. The exact mechanism for the 20 action of these chemotherapeuuc agents are not always known. Despite advances in the field of cancer and leukemia treatments the leading therapies to date are radiation and chemotherapy and bone marrow transplants. Chemotherapeuuc approaches axe said to Sght nancen that are particularly aggressive. Such cytocida1 or cytostauc agents work best on cancers with large growth factors, i.e., ones whose cells are rapidly dividing. To date. 25 hormones, in particular estrogen, progesterone and testosterone, and some anubioucs produced by a variety of microbes, alkylating agents, and anu-metabolites form the bulk of therapies available to oncologists. Ideally cytotoxic agents that have specificity for leukemia, cancer and tumor cells while not affecting normal cells would be extremely desirable. Unfortunately, none have been fijund and instrad agents which target espenally rapidly dividing cells (both diseased and normal) 30 have been used. Clearly, the development of materials that would target cancer or leukemia ceils due to some unique speafiaty for them would be a breakthrough. Alternatively, materials that were cytotoxic to leukemia or cancer cells while exerting mild effects on normal cells would be desirable Therefore, it is an object of an embodiment of this invention to provide a 35 pharmaceutical composition that is effective in treating leukemia with mild effects on normal blood cells * New Zealand Patent Specification No. 315184 (from which the present specification is divided) has relating to the treatment of viral infections WO 9- 058-3 INTELLECTUAL PROPER] OFriCE OF N.Z. 1 3 0 JUL 2001 RECEIVED ^ © PCT7US96/I2474 10 15 -2- More specifically, it is an object of an embodiment of this invention to provide a composition comprising a pharmaceutical earner and a 2-(2,4-difluorophenyl)-l,3-bis(lH-l,2,4-triazol-l-yl)propan-2-ol or a derivative thereof as defined herein suitable for treating cancer, leukemia or tumors The use of 2-<2.4-difluorophenyi)-1.3-bis(lH-1.2,4-tnazoi-l-yl)propan-2-ol or a derivative thereof in combination with other chemotherapeutic agents which are effective m destroying the cancer or tumor is a novel method of treatment 2-(2,4-Difluorophenyl)-l,3-bis(lH-l,2,4-triazol-l-yl)propan-2-ol and its derivatives can also be used to treat viral infections either alone or in the presence of a potentiator SUMMARY OF THE INVENTION The present invention relates to a pharmaceutical composition for treatment of mammals, and in particular, warm blooded animals and humans, which are affected by a tumor or a cancer, for example, leukemia, comprising a pharmaceutical carrier and an effective amount of 2-(2,4-difluorophenyl)-l,3-bis(lH-l,2,4-tnazol-l-yl)propan-2-ol (Fluconazole®) or a derivative thereof 2-(2,4-Difluorophenyl)-1,3-bis( 1H-1,2,4-tnazol-l-yl)propan-2-ol has the formula N N OH I N—CH2-€—CH2-N 20 These compositions can be used to inhibit the growth of leukemia, tumors or cancer cells in humans or by administration of an effective amount either orally, rectaily, topically or parentexally, or intravenously. These compositions do not significantly affect healthy ceils. Potentiators can also be used in combinauon wuh 2-<2,4-difluorophenyi)-l.3-bis(lH-lr2,4-triazoJ-l-yi)propan-2-ol and its derivatives as can chemotherapeutic agentv These compositions are particularly effective against the inflim7a virus. The present invention as claimed consists in use of a safe and effective amount of (1) a member selected from the group consisting of 2-(2,4-difluorophenyl)-1,3-bis( 1H-1,2,4-triazol-I-yl)propan-2-oI, a derivative thereof, and mixtures thereof and (2) a safe and effective amount of a chemotherapeutic agent, m the manufacture of a pharmaceutical composition suitable for the treatment of cancer or tumors in warm blooded mammals WO 9- 058"3 PCI7LS96/I2474 DETAILED DESCRIPTION OF THE INVENTION A. DEFINITIONS: As used herein, the term "comprising" means various components can be conjointly employed in the pharmaceutical composition of this invention. Accordingly, the terms "consisting 5 essentially of" and "consisting or are embodied in the term comprising. As used herein, a "pharmaceutical^ acceptable" component is one that is suitable for use with humans and/or animals without undue adverse side effects (such as toxicity, imtauon, and allergic response) commensurate with a reasonable benefit/risk ratio As used herein, the term "safe and effective amount" refers to the quantity of a 10 component which is sufficient to yield a desired therapeutic response without undue adverse side effects (such as toxicity, irritation, or allergic response) commensurate with a reasonable benefit/risk ratio when used in the manner of this invention. The specific "safe and effective amount" will, obviously, vary with such factors as the particular condition being treated, the physical condition of the patient, the type of mammal being treated, the duration of the treatment, 13 the nature of concurrent therapy (if any), and the specific formulations employed and the structure of the compounds or its derivatives. ^ As used herein, a "2-{2,4-difluorophenyl)-1.3-bis(lH-l,2,4-tnazol-l-yl)propan-2-ol derivative" includes its esters and ethers and its pharmaceutical^ acceptable salts. As used herein, a "pharmaceutical addition salts" are salts of 2-(2,4-<iifluorophenyl)-l,3-20 bis( 1H-1,2,4-tnazoi- 1-yi)propan-2-ol with an organic or inorganic acid. These preferred acid addition salts are chiondcs, bromides, sulfates, nitrates, phosphates, sulfonates, formates, tartrates, maleates. malatcs, citrates, benzoates, salicylates, ascoibates, and the like. As used herein, a "pharmaceutical earner" is a pharmaceutical^ acceptable solvent, suspending agent or vehicle for delivering the anu-Ieukemia agent to the animal or human. The 25 carrier may be liquid or solid and is selected with the planned manner of administration in mind As used herein, "cancer" or "leukemia* refers to all types of cancers or neoplasm or maiigMn* which attack normal healthy blood cells or bone marrow which produces blood ceils which are found in mammals As iitH heron, "viruses" includes viruses which cause diseases in warm blooded animals 30 including HIV, inflnwm rhinoviruses, herpes and the like. v/ As visrd herein. "2-(2,4-difluorophenyl)-1,3 -bis( 1H- 1,2,4-urazol-1 -yl)propan-2-ol and its derivatives" includes esters and ethers as well as addition salts. As used herein "potentiators* are materials such as tnprolidine and its cis-isomer or 1H-Benzimidazole-2-propanoic acid which are used in combination with 2-{2,4-difluorophenyl)-l,3-35 bis(lH-1.2,4-tnazol-l-yi)propan-2-ol and its derivatives. Potentiators can affect the immune system or enhance the effectiveness of the drugs WO 9- 058"3 PCT/US96/12474 -4- As used herein "chemotherapeuuc agents" includes DNA-interacuve Agents, Antimetabolites. Tubulin-Interacuve Agents, Hormonal agents and others, such as Asparaginase or hydroxyurea B. 2-{2,4-DIFLUOROPHENYL)-lJ-BIS(lH-U,4.TIUAZOL-l-YL)PROPAN-2-OL 5 AND ITS DERIVATIVES 2-{2,4-difluoropheny!)-1,3-bis( lH-l,2,4-inazol-1-yl)propan-2-ol and its denvauves the following structure OH I F It is prepared according to the method described in U S. 4,404J16 issued to Richardson 10 (1983). The denvauves include the lower carboxylic aad esters of the pro panel group, for example, acetyl, propanoyl. butyl, pentyl and hexyl esters. Particularly preferred are the esters of carboxylic aads having less than seven carbons, and most preferably propyl esters. Aryl carboxylic aads such as salicylic acid, benzoic aad, and related aads can also be used to esteniy 15 the propanol group. Alkyl ethers having less than 7 carbons are also useful derivatives. The pharma^»nti/r>l addiUon salts are salts of 2-(2,4-difluorcphcnyl)-l,3-t>is(lH-l,2,4-tnazol-l-yl)propan-2-ol with an organic or inorganic acid. These preferred aad addition salts are chlondes, bromides, sulfates, nitrates, phosphates, sulfonates, formates, tartrates, maleates, malates, citrates, hrmratr* salicylates, ascorbates, and the like. 20 These compounds are part of a more generic class of fungiades with the formula: OH I Yi N-CH2-C—CH2-N Y2 k J A W J N N WO 9" 058~3 PCT-1S96/12474 -5- whercin R^is an opuonallv-subsututed alkyl, cycioalkyl, aryl (2,4-dichlorophenyl) or aralkyl group, and Y' and Y~ are =CH- or =N-, and salts or metal complexes and ether or esters thereof. While these materials are active against fungus disease, some have been found to be teratogenic Therefore, those materials which exhibit this property are not useful herein 5 C. CHEMOTHERAPEUTIC AGENTS The chemotherapeuuc agents are generally grouped as DNA-interactive Agents, Antimetabolites, Tubulin-Interactive Agents, Hormonal agents and others such as Asparaginase or hydroxyurea. Each of the groups of chemotherapeuuc agents can be further divided by type of activity or compound. The chemotherapeuuc agents used in combination with 2-{2,4-10 difluorophenyi)- l,3-bis( 1H- 1,2,4-tnazol-1 -yl)propan-2-ol and its denvauves include members of all of these groups. For a detailed discussion of the chemotherapeuuc agents and their method of administration, see Dorr, et aJ, Cancer Chemotherapy Handbook, 2d edition, pages 15-34, Appleton & Lange (Connecucut, 1994) herein incorporated by reference DNA-interactive Agents include the allcylating agents, e g. Cispiaun, Cyclophosphamide, 15 Altretamine; the DNA strand-breakage agents, such as Bleomycin; the intercalating topoisomeiase II inhibitors, e.g., Dacunomycin and Doxorubicin); the nonintercaiating topoisomeiase II inhibitors such as, Etoposide and Teniposide, and the DNA minor groove binder Plicamycin The alkylating agents form covalent chemical adducts with cellular DNA, RNA, and 20 protein molecules and with smaller amino aads, glutathione and similar chemicals. Generally, these alkylating agents react with a nucleophilic atom in a cellular constituent such as an amino, carboxyl, phosphate, sulfhydryl group in nucleic aads, proteins, ammo aads. or glutathione. The mechanism and the role of these alkylating agents m cancer therapy is not well understood Typical alkylating agents include; 25 Nitrogen imiffliT'lv such as Chlorambucil. Cyclophosphamide, Isofamide, Mechiorethamine, Melphalan, Uraal mustard, Azuidine such as Thiotepa methaaesulphonate esters such as Busullan; mtroso ureas, such as Carmusune, Lomusune, Streptozocin, 30 platinum complexes, such as Cispiaun. Carboplaun; bioreductrve aflcyiator, such as Mitomycin, and Procarbazine, Dacaifeazme and Altretamine; DNA strand breaking agents include Bleomycin, DNA topoisomerase II inhibitors include the following: 35 Intercalators. such as Amsacnne. Dacunomycin, Daunorubian, Doxorubicin, Idarubicin, and Mitoxantrone; WO 9" 058-3 PCTL'S96/i:4T4 -6- nonintercalators, such as Etoposide and Teniposide The DNA minor groove binder is Plicamycm The antimetabolites interfere with the production of nucleic acids by one or the other of two major mechanisms. Some of the drugs inhibit production of the deoxynbonucleoside 5 triphosphates that are the immediate precursors for DNA synthesis, thus inhibiting DNA replication Some of the compounds are sufficiently like purines or pynmidines to be able to substitute for them in the anabolic nucleotide pathways. These analogs can then be substituted into the DNA and RNA instead of their normal counterparts. The antimetabolites useful herein include' 10 folate antagonists such as Methotrexate and tnmetrexate pynmidine antagonists, such as Fluorouraol, Fluorodeoxyundine, CB3717, Araciudme, Cytarabine, and Floxundine purine antagonists include Mercaptopunne. 6-Thioguanine, Fludarabine. Pentostaun, sugar modified analogs include Cyatrabme, Fludarabine, 15 ribonucleotide reductase inhibitors include hydroxyurea. Tubulin Interactive agents act by binding to specific sites on tubulin, a protein that polymerizes to fonn cellular microtubules. Microtubules are cnucal cell structure units. Wben the interactive agents bind on the protein, the cell can not form microtubules Tubulin Interactive agents include Vincristine and Vinblastine, both alkaloids and Paclitaxel. 20 Hormonal agents are also useful m the treatment of cancels and tumors. They are used in hormonally susceptible tumors and are usually derived from natural sources. These include: estrogens, conjugated estrogens and Ethinyl Estradiol and Diethylstilbesterol, Chlonnanisea and Idenestrol; progestins such as Hydroxyprogesterone caproate. Medroxyprogesterone, and Megestrol; 25 androgens such as testosterone, testosterone propionate; fiuoxymesterone, mettayltestosterooe; Adrenal corticosteroids are derived from natural adrenal Cortisol or hydrocortisone. They are used of their ana inflammatory benefits as well as the ability of some to inhibit mitotic divisions and to halt DNA synthesis. These compounds include. Prednisone, Dexamethasone, 30 Methyiprednisolone, and Prednisolone. Leutimzing hormone releasing hormone agents or gonadotropm-releasing hormone an*agoni«*« are used primarily the treatment of prostate cancer. These include leuprolide acetate and They prevent the biosynthesis of steroids in the testes. Antihormonal antigens include: 35 antiestrogenic agents such as Tamosifen, antiandrogen agents such as Flutamide . and anuadrenal agents such as Mitotane and Aminogiutethimide. WO 97/05873 PCTT.S96/12474 Hydroxyurea appears to act primarily through inhibition of the enzyme nbonucieoude reductase Asparaginase is an enzyme which converts asparagine to nonfunctional aspartic acid and thus blocks protein synthesis in the tumor 5 Taxol is preferred chemotherapeuuc agent. D. POTENTIATORS The "potentiators" can be any material which improves or increase the efficacy of the pharmaceutical composition or acts on the immune system. One such potentiator is tnprolidine and its cis-isomer which are used m combination with the chemotherapeuuc agents and 2-{2,4-10 difluorophenylW,3-bis(lH-l,2,4-tnazol-l-yl)propan-2-ol and its denvauves Tnprolidine is described in US 5,114,951 (1992) Another potenuator is procodazole, lH-Benzimidazole-2-propanoic acid, [B-{2-benzinudazole) propionic acid, 2-{2-carboxyethyl)benzinudazole, propazol] Procodazole is a non-specific active immunoprotecuve agent against viral and bacterial infecuons and can be used with the composiuons claimed herein It is effective with 2-<2,4-difluorophenyl)-15 l,3-bis(lH-l,2,4-tnazol-l-yl)propan-2-ol and its derivatives alone in treaung cancers, tumors, leukemia and viral infecuons or combined with chemotherapeuuc agents. Propionic acid and its salts and esters can also be used in combination with the pharmaceutical composiuons claimed herein. Antioxidant vitamins such as vitamins A. C and E and beta-carotene can be added to 20 these composiuons. E. DOSAGE Any suitable dosage may be given in the method of the invenuon. The type of compound and the earner and the amount will vary widely depending on the species of the warm blooded '""Ml or body weight, and the type of cancer or tumor or viral infection being treated. 25 Generally a dosage of between about 1 milligram (mg) per kilogram (kg) of body weight and about 1000 mg per kg of body weight is suitable for either the 2<2,4-difluorophenylH,3-bis(lH-I,2,4-triazol-l-yl)propan-2-ol and its denvauves or the chemotherapeuuc agent Preferably from 15 mg to about 800 mg/kg of body weight is used. Generally, the dosage in man is lower than for small warm blooded mammaU such as mice. A dosage unit may comprise a single compound or 30 mixtures thereof with other compounds or other cancer inhibiting compounds. The dosage unit can also compose diluents, extenders, earners, liposomes and the like. The unit may be in solid or gel form such as pills, tablets, capsules and the like or in liquid form suitable for oral, rectal, topical, intravenous injection or parenteral admimstrauon or injection into or around the bone marrow. The range and rauo of 2-{2,4-difluorophenyl)-l,3-bis(lH-l,2,4-tnazol-l-yI)propan-2-ol WO 9--058T3 PCTXS96/124'4 and its denvauves to chemotherapeuuc agent will depend on the t>pe of cancer or tumor being treated and the particular chemotherapeuuc agent F. DOSAGE DELIVERY FORMS The chemotherapeuuc agents, 2-{2,4-<iifluorophenyl)-l.3-bis<lH-1.2,4-tnazoi-l-5 yI)propan-2-oi and its denvauves and, opuonally, the potenuators are typically mixed with a pharmaceuucally acceptable earner This earner can be a solid or liquid and the type is generally chosen based on the type of administrauon being used. The active agent can be coadministered in the form of a tablet or capsule, liposome, as an agglomerated powder or in a Liquid form. Examples of suitable solid earners include lactose, sucrose, geiaun and agar Capsule or tablets 10 can be easily formulated and can be made easy to swallow or chew, other solid forms "kMc granules, and bulk powders. Tablets may contain suitable binders, lubneants, diluents, disintegrating agents, colonng agents, flavonng agents, flow-inducing agents, and melting agents. Examples of suitable liquid dosage forms include soluuons or suspensions in water, pharmaceuucally acceptable £ats and oils, alcohols or other organic solvents, including esters, 15 emulsions, syrups or elixirs, suspensions, soluuons and/or suspensions reconsututed from non-effervescent granules and effervescent preparations reconsututed from effervescent granules Such liquid dosage forms may contain, for example, suitable solvents, preservatives, emulsifying agents, suspending agents, diluents, sweeteners, thickeners, and melting agents. Oral dosage forms optionally contain flavoranls and coloring agents. Parenteral and intravenous forms would also 20 include minerals and other matenals to make them compauble with the type of injection or delivery system chosen. Specific examples of pharmaceutical acceptable earners and excipients that may be used to formulate oral dosage forms of the present invenuon are described in U. S. Pat No 3,903,297 to Robert issued Sept 2, 1975. Techniques and composiuons for making dosage forms useful in 25 the present invention are described in the following references: 7 Modern Pharmaceutics. Chapters 9 aad 10 (Banker & Rhodes. Editors, 1979); Lieberman et al. Pharmaceutical Dosage Forms: Tabtes (1981); and Ansel, Introducnon to Pharmaceutical Dosage Forms 2nd Edition (1976). G. METHOD OF TREATMENT 30 The of treatmentnn be any suitable method which is effective in the treatment of the patTyniaf or tumor type being treated. Treatment may be oral, rectal, topical, parenteral or intravenous adxnimsuauon or by mjecuon into the tumor or cancer. The method of applying an effective amount also vanes depending on the leukemia, cancer, tumor or virus being treated. It is believed that parenteral treatment by intravenous, subcutaneous, or intramuscular 35 application of the 2-{2,4-difluorophenyl)-i 3-bisi lH-1.2.4-tnazol-i-yi)propan-2-ol and its WO 9" 058-3 PCT.1S96/12474 denvauves, formulated with an appropnate earner, addiuonal cancer inhibiung compound or compounds or diluent to facilitate applicauon will be the preferred method of admiiustenng the compounds to warm blooded animals In addiuon to the use of chemotherapeuuc agents and potenuators. 2-(2,4-5 difluorophenyi)-1,3-bis< 1H- 1,2.4-tnazol- l-yl)propan-2-al and its denvauves can be combined with fungicides, herbicides or other anuviral agents. Preferred herbicides and fungicides include carbendazim, fluoconazole, benomyl, glyphosate and propicodazole When the pharmaceutical composiuons are used for treatment of viral infection, they can be combined with other anu-vual agents. 10 ANTI VIRAL EVALUATION WITH HUMAN INFLUENZA VIRUS Female CD (mice Charles River Breeding Laboratones, Portage. MI) 5 to 7 weeks old of age at the time of receipt are used. Mice are approximately 6 to 9 weeks old and weigh approximately 20 to 28 grams at the ume test initiation. All mice used in the study do not vary in age by more than 10 days. The mice are housed 6 per cage with bedding The mice are fed rodent 15 diet 5002 (PML Si Louis Missouri) adlibitum. Fresh water is supplied to the mice adlibitum. Human influenza virus, strain AT2/Taiwan/l/64 is used to challenge the mice. The organism is stored at approximately -70°C. Pnor to infectious challenge a vial of frozen stock is thawed and diluted to the appropnate concentiauon in buffered saline soluuon. The mice are ancahcti7cri with Halothane and the virus challenge dose is administered mtra-nasally tn volume 20 of 50 microlitres. Test material* are administered at the concentration and volume as provided below On days 1 through 14, 10 mice per group receive the test articles by oral lavage. Saline control ammaU (10) receive a comparable volume of saline as compared to the test article-dosed mice. Test articleis accomplished at approximately 24 hour intervals. On day 0 approximately 4 25 hours after the dosing of test articles or saline, all mice are challenged mtra-nasally with an infective dose of virus calculated to produce approximately 90% lethality Animals are observed daily for 21 days after infectious challenge for mortality or monbundity. TEST MATERIAL DOSE (mg/kf) PERCENT MORTALITY Fluconazole 350 o Fluconazole 700 30% ^aliiv . 100% Amantadine 75 0% 30 IN VITRO HUMAN TUMOR COLONY FORMING UNITS TEST Solid tumors removed from pauena are minced into 2 to 5 mm fragments and immediately placed in McCoy's Medium 5A plus 10% heat inactivated newborn calf serum plus </div>

Claims

<div class="application article clearfix printTableText" id="claims"> 9"'0S8~3 PCTTS96/12474 -10- 1% penicillin/streptomycin Wuhan 4 hours, these solid tumors are mechanically disassociated with scissors, forced through No 100 stainless steel mesh, through 25 gauge needles, and then washed with McCoys medium as described above. Ascitic, pleural, pericardial fluids and bone marrow are obtained by standard techniques. The fluid or marrow is placed in sterile containers containing 10 units of preservative free heparin per ml. of malignant fluid or marrow After centnfugauon at 150 x g for 10 minutes, the ceils are harvested and washed with McCoy's medium plus 10% heat inactivated calf serum. The viability of cell suspensions is determined on a hemocytometer with trypan blue. Cells to be cloned are suspended in 0 3% agar in enriched CMRL1066 supplemented with 15% heat inactivated horse serum, penicillin (100 units/ml), streptomycin (2mg/ml), glutamine (2mM), insulin (3 units/ml), asparagme (0 6 mg/ml), and HEPES buffer (2mM). For the continuous exposure test each compound is added to the above mixture. Cells are placed in 35 mm petri dishes in a top layer of agar over an underlayer of agar to prevent growth of fibroblasts. Three plates are prepared for each data point The plates are placed in a 37°C incubator, and are removed on day 14 for counting of the number of colonies in each plate. The number of colonies (defined as 50 cells) formed in the 3 compound treated plates is compared to the number of colonies fonned in the 3 control plates, and the percent colonies surviving at the concentration of compound can be tabulated Three positive control plates are used to determine survival rate. Orthosodium vanadate at 200 ng/ml is used as the positive control. If there is <30% cells in the positive control when compared to the untreated control, the test is evaluated. At concentrations of 0.5 and 5.0 ng/ml in a continuous exposure experiment or single dose experiment Fluconazole was not effective (0/3 and 0/13 respectively) against tumors. At concentration of 50.0 iig/tnl in a continuous exposure experiment Fluconazole was effective against lung, non-small cell, and ovarian cancers particularly Over all 4/13 had £50% survival. 'fO ^T? (f\ O 4 j 0 ^ ^ ^ -11- WHAT WE CLAIM IS:

1. Use of a safe and effective amount of (1) a member selected from the group consisting of 2-(2,4-

difluorophenyl)-l,3-bis(lH-l,2,4-triazol-l-yl)propan-2-ol, a derivative thereof, and mixtures thereof and (2) a safe and effective amount of a chemotherapeutic agent, in the manufacture of a pharmaceutical composition suitable for the treatment of cancer or tumors in warm blooded mammals.

2. Use according to Claim 1 wherein said pharmaceutical composition comprises a sufficient amount to deliver from 1 mg to 1000 mg of said 2-{2,4-difluorophenyl)-1»3-bis(1H-1,2,4-triazol-1-yl)propan-2-ol or a derivative thereof per kg body weight to said warm blooded mammal being treated for cancer or tumors.

3. Use according to Claim 1 wherein said chemotherapeutic agent is selected from the group consisting of DNA-interactive Agents, Antimetabolites, Tubulin-lnteractive Agents, and Hormonal agents.

1 intellectual property office of n.z.

3 0 Jul 2001 RECEIVED

4. Use according to Claim 3 wherein said chemotherapeutic agent is selected from the group consisting of Asparaginase, hydroxyurea, Cisplatin, Cyclophosphamide, Altretamine, Bleomycin, Dactinomycin, Doxorubicin, Etoposide, Teniposide and Pticamycin.

5. Use according to Claim 3 wherein said chemotherapeutic agent is selected from the group consisting of Taxol, Methotrexate, Fluorouracil, Fluorodeoxyuridine, CB3717, Azacitidine, Cytarabine, Floxundine, Mercaptopurine, 6-Thioguanine, Fludarabine, and Pentostatin.

6- Use according to Claim 1, wherein said pharmaceutical composition further comprises a potentiator

7. Use according to Claim 1 wherein said pharmaceutical composition comprises a sufficient amount to deliver from 2 mg to 1000 mg of said 2-<2,4-difluorophenyl)-1,3-bis(1H-1,2,4-triazol-1-yl)propan-2-ol or a derivative thereof and from 0.5mg to 40 mgof said chemotherapeutic agent per kg body weight to said warm blooded mammal being treated for cancer or tumors.

8. Use according to Claim 7 wherein said pharmaceutical composition is suitable for oral or enteric, intravenous, or peritoneal administration.

9. Use according to Claim 7, wherein said pharmaceutical composition further comprises a potentiator.

10. Use according to Claim 9, wherein said potentiator is procodazole.

11. Use according to Claim 1 and substantially as herein described with reference to any embodiment disclosed.

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