WO1995024894A2 - Use of lipoxygenase inhibitors as anti-cancer therapeutic and intervention agents - Google Patents

Use of lipoxygenase inhibitors as anti-cancer therapeutic and intervention agents Download PDF

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WO1995024894A2
WO1995024894A2 PCT/US1995/003337 US9503337W WO9524894A2 WO 1995024894 A2 WO1995024894 A2 WO 1995024894A2 US 9503337 W US9503337 W US 9503337W WO 9524894 A2 WO9524894 A2 WO 9524894A2
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derivative
epithelial cell
cancer
subject
lipoxygenase
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PCT/US1995/003337
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French (fr)
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WO1995024894A3 (en
WO1995024894A9 (en
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James L. Mulshine
Marti Jett
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The Government Of The United States Of America, Represented By The Secretary, Department Of Health And Human Services
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Priority to EP95916100A priority Critical patent/EP0750496A1/en
Priority to US08/704,569 priority patent/US6071949A/en
Priority to AU22722/95A priority patent/AU698313B2/en
Publication of WO1995024894A2 publication Critical patent/WO1995024894A2/en
Publication of WO1995024894A9 publication Critical patent/WO1995024894A9/en
Publication of WO1995024894A3 publication Critical patent/WO1995024894A3/en

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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
    • 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
    • A61K31/045—Hydroxy compounds, e.g. alcohols; Salts thereof, e.g. alcoholates
    • A61K31/05—Phenols
    • 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
    • A61K31/12—Ketones
    • 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
    • A61K31/38—Heterocyclic compounds having sulfur as a ring hetero atom
    • 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
    • A61K31/40—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil
    • 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
    • A61K31/40—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil
    • A61K31/403—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil condensed with carbocyclic rings, e.g. carbazole
    • A61K31/404—Indoles, e.g. pindolol
    • A61K31/405—Indole-alkanecarboxylic acids; Derivatives thereof, e.g. tryptophan, indomethacin
    • 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
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/47—Quinolines; Isoquinolines
    • A61K31/475—Quinolines; Isoquinolines having an indole ring, e.g. yohimbine, reserpine, strychnine, vinblastine
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents

Definitions

  • This invention is in the field of the prevention and treatment of cancer. More specifically, this invention relates to the use of 5-lipoxygenase inhibitors or derivatives thereof in preventing and treating cancer.
  • Arachidonic acid is a 20-carbon polyunsaturated fatty acid derived from dietary sources. Oxygenated AA metabolites participate in a variety of biologic and pathological processes including inflammation, bronchial asthma, and shock. Activation of AA metabolism is initiated by the release of AA from the phospholipid pool by the action of enzymes or other mediators. The released AA can be metabolized by either the lipoxygenase pathway where AA is converted by fatty acid lipoxygenases into hydroperoxy derivatives, giving rise to leukotrienes, or the cycloxygenase pathway where a fatty acid transforms AA rapidly to the prostaglandin PGG 2 , which in turn is further converted enzymatically.
  • FLAP 5-lipoxygenase activating protein
  • FLAP appears to be necessary for cellular leuketriene synthesis (U.S. Patent No. 5,182,367; Miller et al. (1990) Nature 343:278-281).
  • FLAP appears to be involved in the translocation of 5-lipoxygenase from the cytosol to the membrane.
  • Other enzymes involved with the sequential metabolism of the lipoxygenase product need to be in close relationship to the 5-lipoxygenase FLAP complex.
  • FLAP and molecules of related structure may be essential to the activity of enzymes in the 5-lipoxygenase pathway as well as the other downstream enzymes required for leuketriene biosynthesis.
  • Inhibitors of the AA 5-lipoxygenase pathway have shown promise in the treatment of inflammation, asthma, and shock, with minimal side effects in pre-clinical and clinical trials (Batt, Prog. Med. Che . 29:1-63 (1992); Larson, Ann. Pharmacother. 27:898-903 (1993)). However, their use for treating or preventing cancer has never been described.
  • the present invention is directed to the use of inhibitors of 5-lipoxygenase functional activity for the treatment and prevention of epithelial cell-derived cancers.
  • the present invention is also directed to the use of inhibitors of other enzymes in the lipoxygenase pathway involved in the metabolism of AA for use in the treatment and prevention of epithelial cell-derived cancers.
  • the present invention relates to the use of lipoxygenase inhibitors in therapeutic applications, in particular to the prevention and treatment of epithelial cell-derived cancers.
  • FIG. 1(a) classic SCLC cell lines NCI-H209, NCI-H345, and NCI-H510.
  • Fig. Kb) variant SCLC cell lines NCI-N417 and NCI-H82.
  • Fig. 1(c) NSCLC cell lines NCI-H23, A549, and NCI-H1155.
  • Fig. 2 (a) classic SCLC cell lines NCI-H209 and NCI-H345.
  • Fig. 2(b) variant SCLC cell lines NCI-N417 and NCI-H82.
  • Fig. 2(c) NSCLC cell lines A549 and NCI- H1155.
  • Fig. 3 (a) classic SCLC cell lines NCI-H209 and NCI-H345.
  • Fig. 3 (b) variant SCLC cell lines NCI- N417 and NCI-H82.
  • Fig. 3(c) NSCLC cell lines A549 and 5 NCI-H1155.
  • FIG. 1 Structures of MK-886 and related structures: L-669,572, L-663,511, L-665,210, L-654,639, L-668,017 and MK-591.
  • the present invention provides a method for 5 treating or preventing an epithelial cell-derived cancer in a subject in need of such treatment or prevention which comprises administering to the subject an amount of a 5- lipoxygenase inhibitor or derivative thereof, effective to treat or prevent the epithelial cell-derived cancer.
  • the 0 present invention also provides a method for treating or preventing an epithelial cell derived cancer in a subject in need of such treatment by administering an effective amount of an inhibitor of other enzymes involved in the metabolism of arachidonic acid in the 5-lipoxygenase # pathway which comprises administering to the subject an amount of the inhibitor effective to treat or prevent a epithelial cell derived cancer.
  • treatment includes partial or total inhibition of the cancer growth, as well as partial or total destruction of the cancer cells.
  • prevention includes either preventing the onset of clinically evident cancer altogether or preventing the onset of a preclinically evident stage of cancer in individuals at risk, also intended to encompassed by this definition is the prevention of initiation for malignant cells or to arrest or reverse the progression of premalignant cells to malignant cells.
  • the epithelial cell- derived cancer includes basal cell carcinoma, adenocarcinoma, colon cancer, prostate cancer, renal cell carcinoma, and other known cancers that effect epithelial cells throughout the body.
  • the epithelial cell-derived cancer is lung cancer or breast cancer.
  • subject for purposes of treatment includes any human or animal subject who has any one of the known epithelial cell-derived cancers, and preferably is a human subject.
  • the subject is any human or animal subject, and preferably is a human subject who is at risk for obtaining an epithelium cell-derived cancer.
  • the subject may be at risk due to exposure to carcinogenic agents, being genetically predisposed to have the cancer, and the like.
  • Inhibitors of the 5-lipoxygenase pathway in the metabolism of arachidonic acid used in the prevention and treatment of epithelial cell derived cancers may inhibit enzyme activity through a variety of mechanisms.
  • the inhibitor may block or reverse the association of the enzyme with the membrane or inhibit the ° translocation of specific enzymes such as 5-lipoxygenase via a protein such as FLAP.
  • the inhibitors used in the methods described herein may block the enzyme activity directly by acting as a substrate for the enzyme or by depriving the enzyme of necessary cofactors. 5
  • the methods provided herein relate to the use of
  • the 5-lipoxygenase inhibitor is 2-(12-Hydroxydodeca-5, 10-diynyl)-3,5,6-
  • Derivatives are intended to encompass any compounds which are structurally related to AA861, NDGA or MK886 or which possess the substantially equivalent
  • inhibitors may include, but are not limited to, derivatives which act as co-factor antagonist, better enzymatic substrates or inhibitors of activating peptide function.
  • a derivative of MK886 may have the general formula:
  • R 1 is C ⁇ -C 6 alkyl or OH or RCO- in which R 2 is H or C,-C 6 alkyl or quinoline or a long chain hydrocarbon possessing 1-32 carbon atoms and 0-6 double bonds.
  • long chain hydrocarbons include, but are not limited to, linoleic acid (18:2) or oleic acid (18:1).
  • R 3 is C,-C 6 alkyl or R 4 S- or R 5 SO- or R 6 S0 2 - and which R 4 or R 5 or R 6 may be C,-C 6 alkyl or quinoline or a long chain hydrocarbon possessing 1-32 carbon atoms and 0-6 double bonds.
  • Examples of long chain hydrocarbon include, but are not limited to, linoleic acid (18:2) or oleic acid (18:1).
  • R 3 examples include, but are not limited to, ° t-butyl thio, cyclic propyl methylthio, phenyl sulfonyl, phenyl, methyl, phenylthiol.
  • MK886 intended to be encompassed by this invention include, but are not limited to, L- 669,572, 3-[1-(p-cholorobenzyl)-5-isopropyl-3-cyclo- 5 propylmethylthioindole-2-yl]-2,2-dimethylpropanoic acid; L-663,511 3-[1-(p-cholorobenzyl)-5-isopropyl-3- phenysulfonylindol-2-yl)-2,2-dimethylpropanoic acid, L- 665,210, 3-[l-(p-chlorobenzyl) -5-isopropyl-3- phenysulfonylindol-2-yl)-2,2-dimethylpropanoic aid; L-654- 0 639, 3 [l-(p-chlorobenzyl)-5-methoxy-3-methylindol-2-yl]- 2,2-dimethylpropanoic acid;
  • the MK886 5 derivative is 3-(1-(4-chlorobenzyl)-3-(1-butyl-thio)-5- (quinolin-2-yl-methoxy)-indol-2-yl) -2,2-dimethyl propanoic acid) (MK-591) (Tagari, et al. (1993) Agents Action 40:62- 71).
  • hydroxyurea derivatives are also used as inhibitors of 5-lipoxygenase in the prevention and treatment of epithelial cell derived cancers.
  • hydroxyurea derivatives include, but are not limited to, (N-(l- benzo(b)thien-2-ylethyl)-N-hydroxyurea) (zileuton) (Tagari, et al. (1993) Agents Action 40:62-71 herein 5 incorporated by reference.
  • inhibitors of other enzymes that metabolize arachidonic acid downstream of 5-lipoxygenase may also be used to prevent or treat epithelial cell derived cancers in a 0 subject in need of such treatment in a method which comprises administering to the subject an amount effective to prevent or treat the epithelial cell derived cancer.
  • Such inhibitors may effect the activity of the enzyme 5 either directly by acting as a substrate inhibitor or by depriving the enzyme of a cofactor.
  • the inhibitor may also act by targeting proteins such as FLAP which are responsible for the translocation of the enzymes to the membrane where the enzymes are activated.
  • the compounds utilized in the methods of the present invention may be present in the form of free bases or pharmaceutically acceptable acid addition salts thereof. Examples of suitable acids for salt formation are: methanesulfonic, sulfuric, hydrochloric, phosphoric, acetic, citric, lactic, ascorbic, maleic, and the like.
  • the administration for the above methods may be affected by means known to those skilled in the art such as oral, rectal, topical (including aerosol), intranasal, intravenous, subcutaneous, intramuscular, intrabronchial, intracavitary, or intraperitoneal routes of administration. If the cancer is localized, local administration rather than system administration is preferred. Formulation in a lipid vehicle may be used to enhance bioavailability.
  • the compound may be combined with a sterile aqueous solution which is preferably isotonic with the blood of the recipient.
  • a sterile aqueous solution which is preferably isotonic with the blood of the recipient.
  • Such formulations may be prepared by dissolving solid active ingredient in water containing physiologically compatible substances such as sodium chloride, glycine, and the like, and having a buffered pH compatible with physiological conditions to produce an aqueous solution, and rendering said solution sterile.
  • the formulations may be present in unit or multi-dose containers such as sealed ampoules or vials.
  • the formulation may be presented as capsules, tablets, powders, granules or a suspension, with conventional additives such as lactose, mannitol, corn starch or potato starch; with binders such as crystalline cellulose, cellulose derivatives, acacia, corn starch or gelatins; with disintegrators such as corn starch, potato starch or sodium carboxymethyl-cellulose; and with lubricants such as talc or magnesium stearate.
  • the compound may be formulated with acid-stable, base- labile coatings known in the art which begin to dissolve in the high pH small intestine. Formulation to enhance local pharmacologic effects and reduce systemic uptake are preferred.
  • Formulations suitable for parenteral administration conveniently comprise a sterile aqueous preparation of the active compound which is preferably made isotonic. Preparations for injections may also be formulated by suspending or emulsifying the compounds in non-aqueous solvent, such as vegetable oil, synthetic aliphatic acid glycerides, esters of higher aliphatic acids or propylene glycol.
  • non-aqueous solvent such as vegetable oil, synthetic aliphatic acid glycerides, esters of higher aliphatic acids or propylene glycol.
  • Formulations for topical use include known gels, creams, oils, and the like.
  • the compounds may be formulated with known aerosol exipients, such as saline, and administered using commercially available nebulizers.
  • Formulation in a fatty acid source may be used to enhance biocompatibility. Aerosol delivery is the preferred method of delivery for epithelial cancers of the lung for prevention application.
  • the active ingredient may be formulated into suppositories using bases which are solid at room temperature and melt or dissolve at body temperature.
  • bases include coca butter, glycerinated gelatin, hydrogenated vegetable oil, polyethylene glycols of various molecular weights, and fatty esters of polyethylene stearate.
  • the dosage form and amount can be readily established by reference to known cancer treatment or prophylactic regiments.
  • the dosage for the inhibitors or derivatives thereof may be from about 0.1 ng/kg to about 450 g/kg, most preferred is about 0.5 ng/kg to about 100 mg/kg, and most preferably is about 1 ng/kg to about 10 mg/kg.
  • the serum concentration of the inhibitor may be from about 1 ⁇ g/ml to about 20 ⁇ g/ml.
  • the actual dose will depend upon whether the administration is for treatment or prophylactic purposes, the route of administration, the location of the cancer, as well as the phar acokinetic properties of the individual treated.
  • the dosage will generally be lower if the compounds are administered locally rather than systemically, and for prevention rather than for treatment. Such treatments may be administered as often as necessary and for the period of time judged necessary by the treating physician.
  • the dosage regime or therapeutically effective amount of the inhibitor to be administrated may need to be optimized for each individual.
  • the administration of the present invention may be for either prevention or treatment purposes.
  • the methods and compositions used herein may be used alone or in conjunction with additional therapies known to those skilled in the art in the prevention or treatment of cancer.
  • the methods and compositions described herein may be used as adjunct therapy.
  • the 5-lipoxygenase inhibitor may be administered alone or in conjunction with other antineoplastic agents or other growth inhibiting agents or other drugs or nutrients.
  • the 5- lipoxygenase inhibitor or derivatives thereof and the inhibitor of other downstream enzymes involved in the metabolism of arachidonic acid may be administered in combination with each other.
  • molecular methods may be used to inhibit the 5-lipoxygenase.
  • antisense constructs generated by methods known to those skilled in the art can be used to target the messenger RNA of the 5-lipoxygenase enzyme.
  • All books, articles or patents reference herein are incorporated by reference.
  • the present invention is described in the following Experimental Details section, which sets forth specific examples to aid in an understanding of the invention, and should not be construed to limit in any way the invention as defined in the claims which follow thereafter.
  • Example 1 Materials and Methods Cell lines: The following cell lines were used: classic small cell lung cancer (SCLC) cell lines NCI-H209, NCI-H345, and NCI-H510; variant SCLC cell lines NCI-N417 and NCI-H82; large cell carcinoma cell line NCI-H1155; adeno carcinoma cell line NCI-H23; and bronchioalveolear carcinoma cell line A549, breast cancer cell line MCF-7 (American Type Tissue Culture Rockville MD; ATCC) and a colon cancer cell line NCI-H630 (ATCC) .
  • SCLC classic small cell lung cancer
  • NCI-N417 and NCI-H82 variant SCLC cell lines NCI-N417 and NCI-H82
  • large cell carcinoma cell line NCI-H1155 adeno carcinoma cell line NCI-H23
  • bronchioalveolear carcinoma cell line A549 breast cancer cell line MCF-7 (American Type Tissue Culture Rockville MD; ATCC) and a colon cancer cell line
  • All cells were grown in RPMI-1640, supplemented with 5% fetal bovine serum (FBS) , penicillin and streptomycin (Gibco, Grand Island, NY) , and were maintained in a 5% C0 2 atmosphere at 37°C. All cell lines were free of mycoplasma contamination.
  • FBS fetal bovine serum
  • penicillin and streptomycin Gibco, Grand Island, NY
  • Biochemical compounds General lipoxygenase inhibitor Nordihydroguaiaretic acid (NDGA) and 5- lipoxygenase inhibitor AA861, are readily commercially available and were purchased from Biomol Research Laboratories (Plymouth Meeting, PA) .
  • 5-lipoxygenase inhibitor MK886 and MK-591 were obtained from Merck Frost. MK886 may be synthesized as described in Gillard, J. , et al. Can. J. Phvsiol. Pharmacol. 67:456-464 (1989)). ° Growth studies: A modification (Promega
  • AA861 was tested in a dose range of 0.5-10 ⁇ M on three classic SCLC cell lines, two variant SCLC cell lines, and three NSCLC cell lines.
  • the effect of AA861 on the in vitro growth of the specific lung cancer cell lines 0 is presented in Figures 1(a), 1(b), and 1(c).
  • AA861 significantly inhibited growth on all of the SCLC cell lines, and two out of three of the NSCLC cell lines.
  • a bronchioalveolear carcinoma, A549 was repeatedly resistant to AA861. The reason for this resistance is 5 unclear.
  • NDGA also was tested on four SCLC cell lines and two NSCLC cell lines, and the results are presented in Figures 2(a), 2(b), and 2(c).
  • NDGA inhibited the growth of all of the lung cancer cell lines tested. MK886 also was tested on four SCLC cell lines and two NSCLC cell 0 lines, and the results are presented in Figures 3(a), 3(b), and 3(c). MK886 showed similar inhibition results as NDGA. AA861 inhibited growth of MCF-7 cells relative to controls (absence of AA861) to 70% and H630 to 50%. NDGA inhibited growth of MCF-7 cells relative to controls 5 80% and H630 cells 50% relative to controls. MK886 inhibited growth of MCF7 cells 100% relative to controls.
  • the therapeutic potential of the 5-lipoxygenase inhibitor, NDGA was assessed in a heterotransplant animal model. Using conventional methodology small cell lung cancer cells (cell line NCI-209) were transplanted into nude mice. Experimental animals received a 0.1% solution
  • 25 derivatives thereof may be also be tested in the heterotransplant model. Suggested serum concentration of the inhibitor in experimental animals may be from about 1 ⁇ g/ml to about 20 ⁇ g/ml. In addition to treatment models, in vivo models of cancer prevention may also be assessed for the therapeutic potential of the inhibitors or

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Abstract

The present method provides a method for treating an epithelial cell-derived cancer in a subject in need of such treatment which comprises administering to the subject an amount of a 5-lipoxygenase inhibitor or derivative thereof effective to treat the epithelial cell-derived cancer. The present invention also provides a method for preventing an epithelial cell-derived cancer in a subject in need of such prevention which comprises administering to the subject an amount of a 5-lipoxygenase inhibitor effective to prevent the epithelial cell-derived cancer. Suitable 5-lipoxygenase inhibitors useful for the methods of the present invention preferably include 2-(12-Hydroxydodeca-5,10-dinyl)-3,5,6-trimethyl-1,4-benzoquinone and derivatives thereof; Nordihydroguaiaretic acid and derivatives; and 3-[1-(4-chlorobenzyl)-3-t-butyl-thio-t-isopropyl-indol-2-yl]-2-2-dimethylpropanoic acid and derivatives thereof. Also intended to be encompassed by this invention are hydroxyurea derivatives as inhibitors of 5-lipoxygenase inhibitors for use in the prevention and treatment of epithelial cell-derived cancers.

Description

USE OF LIPOXYGENASE INHIBITORS AS ANTI-CANCER THERAPEUTIC AND INTERVENTION AGENTS
Field of the Invention
This invention is in the field of the prevention and treatment of cancer. More specifically, this invention relates to the use of 5-lipoxygenase inhibitors or derivatives thereof in preventing and treating cancer.
Background of the Invention
Arachidonic acid (AA) is a 20-carbon polyunsaturated fatty acid derived from dietary sources. Oxygenated AA metabolites participate in a variety of biologic and pathological processes including inflammation, bronchial asthma, and shock. Activation of AA metabolism is initiated by the release of AA from the phospholipid pool by the action of enzymes or other mediators. The released AA can be metabolized by either the lipoxygenase pathway where AA is converted by fatty acid lipoxygenases into hydroperoxy derivatives, giving rise to leukotrienes, or the cycloxygenase pathway where a fatty acid transforms AA rapidly to the prostaglandin PGG2, which in turn is further converted enzymatically.
A protein designated 5-lipoxygenase activating protein (FLAP) appears to be necessary for cellular leuketriene synthesis (U.S. Patent No. 5,182,367; Miller et al. (1990) Nature 343:278-281). FLAP appears to be involved in the translocation of 5-lipoxygenase from the cytosol to the membrane. Other enzymes involved with the sequential metabolism of the lipoxygenase product need to be in close relationship to the 5-lipoxygenase FLAP complex. FLAP and molecules of related structure may be essential to the activity of enzymes in the 5-lipoxygenase pathway as well as the other downstream enzymes required for leuketriene biosynthesis.
Inhibitors of the AA 5-lipoxygenase pathway have shown promise in the treatment of inflammation, asthma, and shock, with minimal side effects in pre-clinical and clinical trials (Batt, Prog. Med. Che . 29:1-63 (1992); Larson, Ann. Pharmacother. 27:898-903 (1993)). However, their use for treating or preventing cancer has never been described.
The present invention is directed to the use of inhibitors of 5-lipoxygenase functional activity for the treatment and prevention of epithelial cell-derived cancers. The present invention is also directed to the use of inhibitors of other enzymes in the lipoxygenase pathway involved in the metabolism of AA for use in the treatment and prevention of epithelial cell-derived cancers.
Summary of the Invention
In general the present invention relates to the use of lipoxygenase inhibitors in therapeutic applications, in particular to the prevention and treatment of epithelial cell-derived cancers.
It is an object of the present invention to provide a method for treating an epithelial cell-derived cancer in a subject in need of such treatment which comprises administering to the subject an inhibitor of a 5-lipoxygenase enzymatic function or a downstream leuketriene synthetic enzyme effective to treat the epithelial cell-derived cancer.
It is also an object of the present invention to provide a method for preventing an epithelial cell-derived cancer in a subject in need of such prevention which comprises administering to the subject an inhibitor of a 5-lipoxygenase enzymatic function or other downstream leuketriene synthetic enzyme effective to prevent the epithelial cell-derived cancer.
It is yet another object of the present invention to provide a method for preventing an epithelial cell-derived cancer in a subject in need of such prevention which comprises administering to the subject an amount of an inhibitor of an enzyme that metabolizes arachidonic acid effective to prevent an epithelial cell derived cancer.
It is a further object of the present invention to provide a method for treating an epithelial cell derived cancer in a subject in need of such treatment which comprises administering to the subject an amount of an inhibitor of an enzyme that metabolizes arachidonic acid effective to treat the epithelial cell-derived cancer.
It is also another object of the invention to provide a method for treating an epithelial cell derived cancer in a subject in need of such treatment which comprises administering to the subject an amount of a hydroxyurea derivative effective to treat the epithelial cell derived cancer.
It is an object of this invention to provide a method for preventing an epithelial cell derived cancer in a subject in need of such prevention which comprises administering to the subject an amount of a hydroxyurea derivative effective to treat the epithelial cell derived cancer.
It is yet another object of this invention to provide pharmaceutical compositions to use on the methods described herein.
Brief Description of the Figures
Figures Ifa), Kb), and 1(c). Effect of 5-lipoxygenase inhibitor AA861 on the in vitro growth of specific lung ° cancer cell lines. Fig. 1(a) : classic SCLC cell lines NCI-H209, NCI-H345, and NCI-H510. Fig. Kb): variant SCLC cell lines NCI-N417 and NCI-H82. Fig. 1(c): NSCLC cell lines NCI-H23, A549, and NCI-H1155. Figures 2(a) . 2(b) , and 2(c). Effect of 5-lipoxygenase
5 inhibitor NDGA on the In vitro growth of lung cancer cell lines. Fig. 2 (a) : classic SCLC cell lines NCI-H209 and NCI-H345. Fig. 2(b): variant SCLC cell lines NCI-N417 and NCI-H82. Fig. 2(c) : NSCLC cell lines A549 and NCI- H1155. 0 Figures 3(a) , 3(b) , and 3(c). Effect of 5-lipoxygenase inhibitor MK-886 on the in vitro growth of lung cancer cell lines. Fig. 3 (a) : classic SCLC cell lines NCI-H209 and NCI-H345. Fig. 3 (b) : variant SCLC cell lines NCI- N417 and NCI-H82. Fig. 3(c): NSCLC cell lines A549 and 5 NCI-H1155.
Figure 4. Structures of MK-886 and related structures: L-669,572, L-663,511, L-665,210, L-654,639, L-668,017 and MK-591.
Figure 5. Effect on tumor volume over time in animals 0 treated with NDGA (•) and control animals receiving phosphate buffered saline (PBS;0)
Detailed Description of the Invention
The present invention provides a method for 5 treating or preventing an epithelial cell-derived cancer in a subject in need of such treatment or prevention which comprises administering to the subject an amount of a 5- lipoxygenase inhibitor or derivative thereof, effective to treat or prevent the epithelial cell-derived cancer. The 0 present invention also provides a method for treating or preventing an epithelial cell derived cancer in a subject in need of such treatment by administering an effective amount of an inhibitor of other enzymes involved in the metabolism of arachidonic acid in the 5-lipoxygenase # pathway which comprises administering to the subject an amount of the inhibitor effective to treat or prevent a epithelial cell derived cancer.
The term "treatment" includes partial or total inhibition of the cancer growth, as well as partial or total destruction of the cancer cells.
The term "prevention" includes either preventing the onset of clinically evident cancer altogether or preventing the onset of a preclinically evident stage of cancer in individuals at risk, also intended to encompassed by this definition is the prevention of initiation for malignant cells or to arrest or reverse the progression of premalignant cells to malignant cells.
In both methods above, the epithelial cell- derived cancer (epithelial carcinoma) includes basal cell carcinoma, adenocarcinoma, colon cancer, prostate cancer, renal cell carcinoma, and other known cancers that effect epithelial cells throughout the body. Preferably, the epithelial cell-derived cancer is lung cancer or breast cancer.
The term "subject" for purposes of treatment includes any human or animal subject who has any one of the known epithelial cell-derived cancers, and preferably is a human subject. For methods of prevention, the subject is any human or animal subject, and preferably is a human subject who is at risk for obtaining an epithelium cell-derived cancer. The subject may be at risk due to exposure to carcinogenic agents, being genetically predisposed to have the cancer, and the like.
Inhibitors of the 5-lipoxygenase pathway in the metabolism of arachidonic acid used in the prevention and treatment of epithelial cell derived cancers may inhibit enzyme activity through a variety of mechanisms. By the way of example, the inhibitor may block or reverse the association of the enzyme with the membrane or inhibit the ° translocation of specific enzymes such as 5-lipoxygenase via a protein such as FLAP. Alternatively, the inhibitors used in the methods described herein may block the enzyme activity directly by acting as a substrate for the enzyme or by depriving the enzyme of necessary cofactors. 5 The methods provided herein relate to the use of
5-lipoxygenase inhibitors or derivatives thereof in the prevention and treatment of epithelial cell derived cancers. In the preferred embodiments, the 5-lipoxygenase inhibitor is 2-(12-Hydroxydodeca-5, 10-diynyl)-3,5,6-
10 trimethyl-l,4-benzoquinone (AA861) (Yoshimoto, et al.
(1982) Biochemical Biophysica ACTA 713:470-473; Ashida, Y, et al. (1993) Prostoglandins 26(6) :955) or derivatives thereof; Nordihydroguaiaretic acid (NDGA) (Wang, et al. (1991) Mutation Research 261:153-162; Salari, et al.
15 (1984) Prostoglandins Leukotrienes And Medicine 13:53-60) or derivatives thereof; or 3-[l-(4-chlorobenzyl)-3-t- butyl-thio-5-isopropylindol-2-yl]-2, 2-dimethylpropanoic acid (MK886) (Gillard et al. (1989) Can J. Phvsiol. Pharmacol 67:456-464; Rouzer, et al (1990) Journal of
20 Biological Chemistry 265:1436-1442) or derivatives thereof.
Derivatives are intended to encompass any compounds which are structurally related to AA861, NDGA or MK886 or which possess the substantially equivalent
25 biologic activity of AA861, NDGA or MK886. By way of example, such inhibitors may include, but are not limited to, derivatives which act as co-factor antagonist, better enzymatic substrates or inhibitors of activating peptide function.
-Λ MK886 as used herein has the formula:
35
Figure imgf000009_0001
Derivatives of MK886 are also intended to be encompassed by this invention. In a preferred embodiment a derivative of MK886 may have the general formula:
Figure imgf000009_0002
Wherein R1 is Cι-C6 alkyl or OH or RCO- in which R2 is H or C,-C6 alkyl or quinoline or a long chain hydrocarbon possessing 1-32 carbon atoms and 0-6 double bonds. Examples of long chain hydrocarbons include, but are not limited to, linoleic acid (18:2) or oleic acid (18:1). Wherein R3 is C,-C6 alkyl or R4S- or R5SO- or R6S02- and which R4 or R5 or R6 may be C,-C6 alkyl or quinoline or a long chain hydrocarbon possessing 1-32 carbon atoms and 0-6 double bonds. Examples of long chain hydrocarbon include, but are not limited to, linoleic acid (18:2) or oleic acid (18:1).
Examples of R3 include, but are not limited to, ° t-butyl thio, cyclic propyl methylthio, phenyl sulfonyl, phenyl, methyl, phenylthiol.
Derivatives of MK886 intended to be encompassed by this invention include, but are not limited to, L- 669,572, 3-[1-(p-cholorobenzyl)-5-isopropyl-3-cyclo- 5 propylmethylthioindole-2-yl]-2,2-dimethylpropanoic acid; L-663,511 3-[1-(p-cholorobenzyl)-5-isopropyl-3- phenysulfonylindol-2-yl)-2,2-dimethylpropanoic acid, L- 665,210, 3-[l-(p-chlorobenzyl) -5-isopropyl-3- phenysulfonylindol-2-yl)-2,2-dimethylpropanoic aid; L-654- 0 639, 3 [l-(p-chlorobenzyl)-5-methoxy-3-methylindol-2-yl]- 2,2-dimethylpropanoic acid; and L-668,017 described in Rouzer et al. (1990) Journal of Biological Chemistry 265:1436-1442 which is herein incorporated by reference (Figure 5) . In a preferred embodiment, the MK886 5 derivative is 3-(1-(4-chlorobenzyl)-3-(1-butyl-thio)-5- (quinolin-2-yl-methoxy)-indol-2-yl) -2,2-dimethyl propanoic acid) (MK-591) (Tagari, et al. (1993) Agents Action 40:62- 71).
In yet another embodiment of this invention 0 hydroxyurea derivatives are also used as inhibitors of 5-lipoxygenase in the prevention and treatment of epithelial cell derived cancers. Examples of hydroxyurea derivatives include, but are not limited to, (N-(l- benzo(b)thien-2-ylethyl)-N-hydroxyurea) (zileuton) (Tagari, et al. (1993) Agents Action 40:62-71 herein 5 incorporated by reference.
In an alternative embodiment of this invention inhibitors of other enzymes that metabolize arachidonic acid downstream of 5-lipoxygenase, may also be used to prevent or treat epithelial cell derived cancers in a 0 subject in need of such treatment in a method which comprises administering to the subject an amount effective to prevent or treat the epithelial cell derived cancer. Such inhibitors may effect the activity of the enzyme 5 either directly by acting as a substrate inhibitor or by depriving the enzyme of a cofactor. The inhibitor may also act by targeting proteins such as FLAP which are responsible for the translocation of the enzymes to the membrane where the enzymes are activated. The compounds utilized in the methods of the present invention may be present in the form of free bases or pharmaceutically acceptable acid addition salts thereof. Examples of suitable acids for salt formation are: methanesulfonic, sulfuric, hydrochloric, phosphoric, acetic, citric, lactic, ascorbic, maleic, and the like.
The administration for the above methods may be affected by means known to those skilled in the art such as oral, rectal, topical (including aerosol), intranasal, intravenous, subcutaneous, intramuscular, intrabronchial, intracavitary, or intraperitoneal routes of administration. If the cancer is localized, local administration rather than system administration is preferred. Formulation in a lipid vehicle may be used to enhance bioavailability.
For intravenous, intramuscular, subcutaneous, or intraperitoneal administration, the compound may be combined with a sterile aqueous solution which is preferably isotonic with the blood of the recipient. Such formulations may be prepared by dissolving solid active ingredient in water containing physiologically compatible substances such as sodium chloride, glycine, and the like, and having a buffered pH compatible with physiological conditions to produce an aqueous solution, and rendering said solution sterile. The formulations may be present in unit or multi-dose containers such as sealed ampoules or vials.
For oral administration, the formulation may be presented as capsules, tablets, powders, granules or a suspension, with conventional additives such as lactose, mannitol, corn starch or potato starch; with binders such as crystalline cellulose, cellulose derivatives, acacia, corn starch or gelatins; with disintegrators such as corn starch, potato starch or sodium carboxymethyl-cellulose; and with lubricants such as talc or magnesium stearate. If the cancer is localized in the G.I. tract, the compound may be formulated with acid-stable, base- labile coatings known in the art which begin to dissolve in the high pH small intestine. Formulation to enhance local pharmacologic effects and reduce systemic uptake are preferred.
Formulations suitable for parenteral administration conveniently comprise a sterile aqueous preparation of the active compound which is preferably made isotonic. Preparations for injections may also be formulated by suspending or emulsifying the compounds in non-aqueous solvent, such as vegetable oil, synthetic aliphatic acid glycerides, esters of higher aliphatic acids or propylene glycol.
Formulations for topical use include known gels, creams, oils, and the like. For aerosol delivery, the compounds may be formulated with known aerosol exipients, such as saline, and administered using commercially available nebulizers. Formulation in a fatty acid source may be used to enhance biocompatibility. Aerosol delivery is the preferred method of delivery for epithelial cancers of the lung for prevention application.
For rectal administration, the active ingredient may be formulated into suppositories using bases which are solid at room temperature and melt or dissolve at body temperature. Commonly used bases include coca butter, glycerinated gelatin, hydrogenated vegetable oil, polyethylene glycols of various molecular weights, and fatty esters of polyethylene stearate.
The dosage form and amount can be readily established by reference to known cancer treatment or prophylactic regiments. The dosage for the inhibitors or derivatives thereof may be from about 0.1 ng/kg to about 450 g/kg, most preferred is about 0.5 ng/kg to about 100 mg/kg, and most preferably is about 1 ng/kg to about 10 mg/kg. The serum concentration of the inhibitor may be from about 1 μg/ml to about 20 μg/ml. The actual dose will depend upon whether the administration is for treatment or prophylactic purposes, the route of administration, the location of the cancer, as well as the phar acokinetic properties of the individual treated. The dosage will generally be lower if the compounds are administered locally rather than systemically, and for prevention rather than for treatment. Such treatments may be administered as often as necessary and for the period of time judged necessary by the treating physician. One of skill in the art will appreciate that the dosage regime or therapeutically effective amount of the inhibitor to be administrated may need to be optimized for each individual.
The administration of the present invention may be for either prevention or treatment purposes. The methods and compositions used herein may be used alone or in conjunction with additional therapies known to those skilled in the art in the prevention or treatment of cancer. Alternatively, the methods and compositions described herein may be used as adjunct therapy. By way of example, the 5-lipoxygenase inhibitor may be administered alone or in conjunction with other antineoplastic agents or other growth inhibiting agents or other drugs or nutrients. Alternatively the 5- lipoxygenase inhibitor or derivatives thereof and the inhibitor of other downstream enzymes involved in the metabolism of arachidonic acid may be administered in combination with each other. In an alternative embodiment molecular methods may be used to inhibit the 5-lipoxygenase. By way of example antisense constructs generated by methods known to those skilled in the art can be used to target the messenger RNA of the 5-lipoxygenase enzyme. All books, articles or patents reference herein are incorporated by reference. The present invention is described in the following Experimental Details section, which sets forth specific examples to aid in an understanding of the invention, and should not be construed to limit in any way the invention as defined in the claims which follow thereafter.
Example 1 Materials and Methods Cell lines: The following cell lines were used: classic small cell lung cancer (SCLC) cell lines NCI-H209, NCI-H345, and NCI-H510; variant SCLC cell lines NCI-N417 and NCI-H82; large cell carcinoma cell line NCI-H1155; adeno carcinoma cell line NCI-H23; and bronchioalveolear carcinoma cell line A549, breast cancer cell line MCF-7 (American Type Tissue Culture Rockville MD; ATCC) and a colon cancer cell line NCI-H630 (ATCC) . All cells were grown in RPMI-1640, supplemented with 5% fetal bovine serum (FBS) , penicillin and streptomycin (Gibco, Grand Island, NY) , and were maintained in a 5% C02 atmosphere at 37°C. All cell lines were free of mycoplasma contamination.
Biochemical compounds: General lipoxygenase inhibitor Nordihydroguaiaretic acid (NDGA) and 5- lipoxygenase inhibitor AA861, are readily commercially available and were purchased from Biomol Research Laboratories (Plymouth Meeting, PA) . 5-lipoxygenase inhibitor MK886 and MK-591 were obtained from Merck Frost. MK886 may be synthesized as described in Gillard, J. , et al. Can. J. Phvsiol. Pharmacol. 67:456-464 (1989)). ° Growth studies: A modification (Promega
CellTiter 96®, Promega Madison, WI) of the semiautomated colorimetric assay, MTT (Nakanishi, et al. Exper. Cell Biol. 56:74-85 (1988)), which quantitates cell numbers based on reduction of a tetrazolium compound by tumor cells as determined by a spectrophotometer (540 nm) was used. All assays were performed in RPMI-1640 media supplemented with transferrin (T) 10 μg/ml, insulin (I) 5 μg/ml and selenium (S) 5xl08 M (TIS) (Sigma Chemicals, St. Louis, MO) . Seeding densities were 1-2X104 cells/well, 0 and cells were grown for 5 days. Each experiment was reported as mean optical density corrected for background =/- standard deviation for a minimum of six data points per experiment, with each experiment repeated at least three times. Results
AA861 was tested in a dose range of 0.5-10 μM on three classic SCLC cell lines, two variant SCLC cell lines, and three NSCLC cell lines. The effect of AA861 on the in vitro growth of the specific lung cancer cell lines 0 is presented in Figures 1(a), 1(b), and 1(c). AA861 significantly inhibited growth on all of the SCLC cell lines, and two out of three of the NSCLC cell lines. A bronchioalveolear carcinoma, A549, was repeatedly resistant to AA861. The reason for this resistance is 5 unclear. NDGA also was tested on four SCLC cell lines and two NSCLC cell lines, and the results are presented in Figures 2(a), 2(b), and 2(c). NDGA inhibited the growth of all of the lung cancer cell lines tested. MK886 also was tested on four SCLC cell lines and two NSCLC cell 0 lines, and the results are presented in Figures 3(a), 3(b), and 3(c). MK886 showed similar inhibition results as NDGA. AA861 inhibited growth of MCF-7 cells relative to controls (absence of AA861) to 70% and H630 to 50%. NDGA inhibited growth of MCF-7 cells relative to controls 5 80% and H630 cells 50% relative to controls. MK886 inhibited growth of MCF7 cells 100% relative to controls.
Example 2 In Vivo Models of Cancer and Carcinogeneses
The therapeutic potential of the 5-lipoxygenase inhibitor, NDGA, was assessed in a heterotransplant animal model. Using conventional methodology small cell lung cancer cells (cell line NCI-209) were transplanted into nude mice. Experimental animals received a 0.1% solution
10 of NDGA in water ad libitum and drank about 5cc of water/day. Control animals received phosphate buffered saline. The xenografts of the small cell lung cancer line, NCI-209, showed a significant reduction in heterotransplant size (compared to controls) at 2.5 to 4.5
15 weeks after cell line engraftment in mice receiving orally administered NGDA. (see Figure 5 and Table I)
Table I. Weight of nude mice (grams)
Treatment Mouse Tumor
20 None 26.5 ± 1.3 1.29 ± 0.07 NDGA 25.9 ± 1.9 0.60 ± 0.19*
The mean weight ± S.D. of 4 determinations is indicated; p 0.05, * using students t-test.
AA861 and derivatives thereof and MK886 and
25 derivatives thereof may be also be tested in the heterotransplant model. Suggested serum concentration of the inhibitor in experimental animals may be from about 1 μg/ml to about 20 μg/ml. In addition to treatment models, in vivo models of cancer prevention may also be assessed for the therapeutic potential of the inhibitors or
30 derivatives thereof.
All publications mentioned hereinabove are hereby incorporated by reference in their entirety. While the foregoing invention has been described in some detail <35l for purposes of clarity and understanding, it will be appreciated by one skilled in the art from a reading of the disclosure that various changes in form and detail can be made without departing from the true scope of the invention in the appended claims.

Claims

What is Claimed is:
1. A method for treating an epithelial cell- derived cancer in a subject in need of such treatment which comprises administering to the subject an amount of a 5-lipoxygenase inhibitor or derivative thereof effective to treat the epithelial cell-derived cancer.
2. The method of claim 1, wherein the 5- lipoxygenase inhibitor is 2-(12-Hydroxydodeca-5,10- diynyl) -3,5,6-trimethyl-l,4-benzoquinone or derivative thereof.
3. The method of claim 1, wherein the 5- lipoxygenase inhibitor is Nordihydroguaiaretic acid or derivative thereof.
4. The method of claim 1, wherein the 5- lipoxygenase inhibitor is 3-[l-(4-chlorobenzyl) -3-t-butyl- thio-t-isopropyl-indol-2-yl]-2,2-dimethylpropanoic acid or derivative thereof.
5. The method of claim 4 wherein the derivative is selected from the group consisting of L-669,572, L-663,511, L-665,210, L-654,639, L-668,017 and MK-591.
6. The method of claim 5, wherein the derivative is (3-(l-(4 clorobenzyl)-3-(t-butyl-thio)-5- (quinoline-2-yl-methoxy) -indol-2-yl) -2,2-dimethyl propionic acid (MK-591) .
7. The method of claim 1, wherein the 5- lipoxygenase inhibitor is a hydroxyurea derivative.
8. The method of claim 7, wherein the hydroxyurea derivative is zileuton.
9. The method of claim 1, wherein the epithelial cell-derived cancer is lung cancer, colon cancer, or breast cancer.
10. The method of claim 1, wherein the administration is affected by oral, rectal, topical aerosol, intravenous, subcutaneous, intramuscular, intrabronchial, intracavitary, or intraperitoneal administration.
11. The method of claim 1, wherein the amount is between about 1 ng/kg and about 10 mg/kg.
12. A method for preventing an epithelial cell- derived cancer in a subject in need of such prevention which comprises administering to the subject an amount of a 5-lipoxygenase inhibitor or derivative thereof effective to prevent the epithelial cell-derived cancer.
13. The method of claim 12, wherein the 5- lipoxygenase inhibitor is 2-(12-Hydroxydodeca-5,10- diynyl)-3,5,6-trimethyl-l,4-benzoquinone.
14. The method of claim 12 , wherein the 5- 5 lipoxygenase inhibitor is Nordihydroguaiaretic acid or derivative thereof.
15. The method of claim 12, wherein the 5- lipoxygenase inhibitor is 3-[l-(4-chlorobenzyl)-3-t-butyl- 0 thio-t-isopropyl-indol-2-yl]-2-2-dimethylpropanoic acid or derivative thereof.
16. The method of claim 15, wherein the s derivative is selected from the group consisting of L-669,572, L-663,511, L-665,210, L-654,639, L-668,017 and MK-591.
17. The method of claim 16, wherein the derivative is (3-(l-(4 clorobenzyl)-3-(t-butyl-thio)-5- (quinoline-2-yl-methoxy) -indol-2-yl) -2,2-dimethyl propionic acid (MK-591) .
18. The method of claim 1, wherein the 5- lipoxygenase inhibitor is a hydroxyurea derivative.
19. The method of claim 7, wherein the hydroxyurea derivative is zileuton.
20. The method of claim 12, wherein the epithelial cell-derived cancer is lung cancer, colon cancer or breast cancer.
21. The method of claim 12, wherein the administration is affected by oral, rectal, topical aerosol, intravenous, subcutaneous, intramuscular, intrabronchial, intracavitary, or intraperitoneal administration.
22. The method of claim 12, wherein the amount is between about 1 ng/kg and about 10 mg/kg.
23. A use of a 5-lipoxygenase inhibitor or a derivative thereof in a pharmaceutical composition in the treatment of an epithelial cell cancer in a subject.
24. The use claim 23, wherein the 5- lipoxygenase inhibitor is 2-(12-Hydroxydodeca-5,10- diynyl) -3,5,6-trimethyl-l,4-benzoquinone or derivative thereof.
25. The use of claim 23, wherein the 5- lipoxygenase inhibitor is Nordihydroguaiaretic acid or derivative thereof.
26. The use of claim 23, wherein the 5- lipoxygenase inhibitor is 3-[l-(4-chlorobenzyl)-3-t-butyl- thio-t-isopropyl-indol-2-yl]-2, 2-dimethylpropanoic acid or derivative thereof.
27. The use of Claim 26, wherein the 5- lipoxygenase inhibitor is selected from the group consisting of L-669,572, L-663,511, L-665,210, L-654,639, L-668,017 and MK-591.
28. The use of claim 27, wherein the derivative is (3-(l-(4 clorobenzyl)-3-(t-butyl-thio)-5-(quinoline-2- yl-methoxy)-indol-2-yl)-2,2-dimethyl propionic acid (MK- 591) .
29. The use of claim 23, wherein the derivative is a hydroxyurea derivative.
30. The use of claim 29, wherein the hydroxyurea derivative is zileuton.
31. The use of claim 23, wherein the epithelial cell-derived cancer is lung cancer, colon cancer or breast cancer.
32. The use of claim 23, wherein the administration is affected by oral, rectal, topical aerosol, intravenous, subcutaneous, intramuscular, intrabronchial, intracavitary, or intraperitoneal administration.
33. The use of claim 23, wherein the amount of the 5-lipoxygenase inhibitor is in the range of about 1 ng/kg and about 40 mg/kg.
34. The use of claim 33, wherein the amount of the 5-lipoxygenase inhibition is in the range of about 1 ng/kg to about 10 mg/kg.
35. A method for treating an epithelial cell derived cancer in a subject which comprises administering to the subject an amount of an inhibitor of an enzymes that metabolizes arachidonic acid effective to treat the epithelial cell derived cancer.
36. A method for preventing an epithelial cell derived cancer in a subject which comprises administering to the subject an amount of an inhibitor of an enzyme that metabolizes arachidonic acid effective to treat the epithelial cell derived cancer.
37. A use of an inhibitor of an enzyme that metabolizes arachidonic acid in a pharmaceutical composition in the treatment or prevention of an epithelial cell derived cancer in a subject.
38. A method for treating an epithelial cell derived cancer in a subject which comprises administering to the subject an amount of an inhibitor of a 5- lipoxygenase downstream enzyme that metabolizes arachidonic acid effective to treat the epithelial cell derived cancer.
39. A method for preventing an epithelial cell derived cancer in a subject which comprises administering to the subject an amount of an inhibitor of a 5- lipoxygenase downstream enzyme that metabolizes arachidonic acid effective to treat the epithelial cell derived cancer.
40. A use of an inhibitor of a 5-lipoxygenase downstream enzyme that metabolizes arachidonic acid in a pharmaceutical composition in the treatment or prevention of an epithelial cell derived cancer in a subject.
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