WO2010005851A1 - Combination therapy for treating iron disorders - Google Patents

Combination therapy for treating iron disorders Download PDF

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Publication number
WO2010005851A1
WO2010005851A1 PCT/US2009/049443 US2009049443W WO2010005851A1 WO 2010005851 A1 WO2010005851 A1 WO 2010005851A1 US 2009049443 W US2009049443 W US 2009049443W WO 2010005851 A1 WO2010005851 A1 WO 2010005851A1
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methyl
methylene
dicarbamimidothioate
bis
isothiourea
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PCT/US2009/049443
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French (fr)
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Robin Paul Sherrington
Alison Jane Cutts
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Xenon Pharmaceuticals Inc.
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Publication of WO2010005851A1 publication Critical patent/WO2010005851A1/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
    • A61K31/13—Amines
    • A61K31/155—Amidines (), e.g. guanidine (H2N—C(=NH)—NH2), isourea (N=C(OH)—NH2), isothiourea (—N=C(SH)—NH2)
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/16—Amides, e.g. hydroxamic acids
    • 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
    • A61K31/41—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with two or more ring hetero atoms, at least one of which being nitrogen, e.g. tetrazole
    • A61K31/4196—1,2,4-Triazoles
    • 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
    • A61K31/41—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with two or more ring hetero atoms, at least one of which being nitrogen, e.g. tetrazole
    • A61K31/425—Thiazoles
    • A61K31/426—1,3-Thiazoles
    • 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
    • A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
    • A61K31/4412—Non condensed pyridines; Hydrogenated derivatives thereof having oxo groups directly attached to the heterocyclic ring
    • 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
    • A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
    • A61K31/4427—Non condensed pyridines; Hydrogenated derivatives thereof containing further heterocyclic ring systems
    • A61K31/4439—Non condensed pyridines; Hydrogenated derivatives thereof containing further heterocyclic ring systems containing a five-membered ring with nitrogen as a ring hetero atom, e.g. omeprazole
    • 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
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00

Definitions

  • the present invention is directed to combination therapies for the treatment of iron overload disorders utilizing inhibitors of divalent metal transporter-1 (DMT1 ), proton pump inhibitors (PPIs), iron chelators and calcium channel blockers.
  • DMT1 divalent metal transporter-1
  • PPIs proton pump inhibitors
  • iron chelators iron chelators and calcium channel blockers.
  • Iron overload Defects in iron metabolism contributing to iron overload represent a chronic health problem in adults. Iron overload is observed in diseases such as hemochromatosis, thalassemia and certain forms of anemia. In addition, iron overload can be caused by repeated blood transfusions outside of the treatment of these disorders. Iron overload can result in cirrhosis, hepatoma, arthritis, hypogonadism, diabetes and cardiomyopathy. Current therapies for iron overload include phlebotomy and treatment with chelators. However, given safety, side effects and compliance issues with both of these treatments, a market exists for safe drugs or combination therapies that act to block iron absorption in the gut and increase iron excretion.
  • iron chelators such as Deferasirox, Deferoxamine and Deferiprone
  • the efficacy of these iron chelators can be limited by dose dependent severe side effects particularly when administered at the higher dosages. These side effects can include, for example, renal impairment, liver toxicity and cytopenias. If these side effects are observed it is recommended that the chelation therapy be discontinued or the daily dosage of the chelator be reduced. This results in less efficacy as measured by the amount of iron removed from the body.
  • combination therapy wherein two or more chelators are administered, has been proposed.
  • omeprazole resulted in a 50% decrease in ferrous iron absorption in rats on an iron- deficient diet (see, Golubov, J. et a/., Dig. Dis. Sci. (1991 ), Vol. 36, No. 4, pp. 405-408).
  • the dose of omeprazole employed produced an effect on gastric acid output in rats similar to that produced by a dose recommended for the treatment of human gastrointestinal diseases.
  • proton pump inhibitors e.g. lansoprazole, esomeprazole, omeprazole and pantoprazole
  • proton pump inhibitors e.g. lansoprazole, esomeprazole, omeprazole and pantoprazole
  • lansoprazole, esomeprazole, omeprazole and pantoprazole increased the risk of heart attacks and strokes by 50 percent among cardiac patients taking Plavix (see, Norgard , N. et a/., Ann. Pharmacother. (2009), published on-line on 2009 May 26). Therefore, it would appear that lengthy exposure to clinical dosages of proton pump inhibitors has serious liabilities.
  • Other long term side effects of proton pump inhibitors include increased risks of colon cancer, gastric cancer, infection with C. difficile, respiratory infections and bone fracture.
  • nifedipine dimethyl 1 ,4-dihydro-2,6-dimethyl-4-(o-nitrophenyl)-3,5- pyridinedicarboxylate
  • nifedipine dimethyl 1 ,4-dihydro-2,6-dimethyl-4-(o-nitrophenyl)-3,5- pyridinedicarboxylate
  • DMT1 -mediated cellular iron transport and enhances urinary iron excretion (see, Ludwiczek, et al., (2007), Vol. 13, No. 4, pp. 448-454).
  • the mechanism by which nifedipine causes this effect is by prolonging the iron transporting activity of DMT1 in the duodenum and thus likely increase urinary iron output by prolonging the iron transporting activity of DMT1 in the kidney.
  • DMT1 is expressed in the duodenum and regulates absorption of iron. Recently, a series of compounds has been developed as DMT1 inhibitors (see, for example, PCT Published Application No. WO 2008/109840; PCT Published Application No. WO 2008/115999; PCT Published Application No. WO 2008/118790; PCT
  • iron chelators in combination with a proton pump inhibitor (PPI), a calcium channel blocker or a DMT1 inhibitor, or any combination thereof, is not known.
  • PPI proton pump inhibitor
  • a PPI, a DMT1 inhibitor or a calcium channel blocker could work additively with chelation therapy to remove the excess iron that is in the body by blocking the uptake of iron from the intestine into the body or by increasing the urinary iron output from the kidney.
  • This combination may increase efficacy and improve iron serum and tissue control in iron-overloaded patients or could lead to reduction in the dosage or frequency of dosing chelators which could lead to safer treatments for patients by reducing the chelation treatment-induced side effects.
  • iron overload disorders preferably primary iron overload and transfusional iron overload in mammals, preferably in humans, using a combination of DMT1 inhibitors with iron chelators, calcium channel blockers or PPIs, or any combination thereof, or PPIs with iron chelators.
  • the present invention is directed to combination therapies for treating an iron overload disorder in a mammal, wherein the combination therapies comprise administering to the mammal a therapeutically effective amount of a DMT1 inhibitor and a therapeutically effective amount of a proton pump inhibitor, and, optionally, a therapeutically effective amount of a pharmaceutically acceptable iron chelator, and, optionally a therapeutically effective amount of a calcium channel blocker.
  • the present invention is directed to combination therapies for treating an iron overload disorder in a mammal, wherein the combination therapies comprise administering to the mammal a therapeutically effective amount of a DMT1 inhibitor and a therapeutically effective amount of a pharmaceutically acceptable iron chelator and, optionally, a therapeutically effective amount of a calcium channel blocker.
  • the present invention is directed to combination therapies for treating an iron overload disorder in a mammal, wherein the combination therapies comprise administering to the mammal a therapeutically effective amount of a DMT1 inhibitor and a therapeutically effective amount of a calcium channel blocker, and, optionally, a therapeutically effective amount of a proton pump inhibitor.
  • the present invention is directed to combination therapies for treating an iron overload disorder in a mammal, wherein the combination therapies comprise administering to the mammal a therapeutically effective amount of a pharmaceutically acceptable iron chelator and a therapeutically effective amount of a calcium channel blocker and/or a therapeutically effective amount of a proton pump inhibitor.
  • the present invention is directed to combination therapies for treating an iron overload disorder in a mammal, wherein the combination therapies comprise administering to the mammal a therapeutically effective amount of a proton pump inhibitor and a therapeutically effective amount of a calcium channel blocker.
  • this invention is directed to a method of treating an iron overload disorder in a mammal, wherein the method comprises administering to the mammal in need thereof a therapeutically effective amount of a DMT1 inhibitor and a therapeutically effective amount of a pharmaceutically acceptable iron chelator.
  • this invention is directed to a method of treating an iron overload disorder in a mammal, wherein the method comprises administering to the mammal in need thereof a therapeutically effective amount of a DMT1 inhibitor and a therapeutically effective amount of a proton pump inhibitor.
  • this invention is directed to a method of treating an iron overload disorder in a mammal, wherein the method comprises administering to the mammal in need thereof a therapeutically effective amount of a DMT1 inhibitor and a therapeutically effective amount of a calcium channel blocker.
  • this invention is directed to a method of treating an iron overload disorder in a mammal, wherein the method comprises administering to the mammal in need thereof a therapeutically effective amount of a DMT1 inhibitor, a therapeutically effective amount of a pharmaceutically acceptable iron chelator, and a therapeutically effective amount of a proton pump inhibitor.
  • this invention is directed to a method of treating an iron overload disorder in a mammal, wherein the method comprises administering to the mammal in need thereof a therapeutically effective amount of a DMT1 inhibitor, a therapeutically effective amount of a pharmaceutically acceptable iron chelator, and a therapeutically effective amount of a calcium channel blocker.
  • this invention is directed to a method of treating an iron overload disorder in a mammal, wherein the method comprises administering to the mammal in need thereof a therapeutically effective amount of a DMT1 inhibitor, a therapeutically effective amount of a proton pump inhibitor, and a therapeutically effective amount of a calcium channel blocker.
  • this invention is directed to a method of treating an iron overload disorder in a mammal, wherein the method comprises administering to the mammal in need thereof a therapeutically effective amount of a DMT1 inhibitor, a therapeutically effective amount of a calcium channel blocker, a therapeutically effective amount of a proton pump inhibitor, and a therapeutically effective amount of a pharmaceutically acceptable iron chelator.
  • this invention is directed to a method for treating an iron overload disorder in a mammal, wherein the combination therapy comprises the administration to the mammal in need thereof of a therapeutically effective amount of a proton pump inhibitor and a therapeutically effective amount of a pharmaceutically acceptable iron chelator.
  • this invention is directed to a method for treating an iron overload disorder in a mammal, wherein the combination therapy comprises the administration to the mammal in need thereof of a therapeutically effective amount of a proton pump inhibitor, a therapeutically effective amount of a pharmaceutically acceptable iron chelator and a therapeutically effective amount of a calcium channel blocker.
  • this invention is directed to a method for treating an iron overload disorder in a mammal, wherein the combination therapy comprises the administration to the mammal in need thereof of a therapeutically effective amount of a proton pump inhibitor and a therapeutically effective amount of a calcium channel blocker.
  • C 7 -C 12 alkyl describes an alkyl group, as defined below, having a total of 7 to 12 carbon atoms
  • C 4 -C 12 cycloalkylalkyl describes a cycloalkylalkyl group, as defined below, having a total of 4 to 12 carbon atoms.
  • the total number of carbons in the shorthand notation does not include carbons that may exist in substituents of the group described.
  • Amino refers to the -NH 2 radical.
  • Niro refers to the -NO 2 radical.
  • Thifluoromethyl refers to the -CF 3 radical.
  • Alkyl refers to a straight or branched hydrocarbon chain radical consisting solely of carbon and hydrogen atoms, containing no unsaturation, having from one to twelve carbon atoms, preferably one to eight carbon atoms or one to six carbon atoms, and which is attached to the rest of the molecule by a single bond, e.g., methyl, ethyl, n-propyl, 1-methylethyl (/so-propyl), n-butyl, n-pentyl, 1 ,1-dimethylethyl (f-butyl),
  • an alkyl group may be optionally substituted by one of the following groups: alkyl, alkenyl, halo, haloalkenyl, cyano, nitro, aryl, cycloalkyl, heterocyclyl, heteroaryl, oxo, trimethylsilanyl, -OR 30 , -OC(O)-R 30 , -N(R 30 ) 2> -C(O)R 30 , -C(O)OR 30 , -C(O)N(R 30 ) 2 , -N(R 30 )C(O)OR 32 , -N(R 30 )C(O)R 32 , -N(R 30 )S(O),R 32 (where t is 1 to 2), -S(O) 1 OR 32 (where t is 1 to 2), -S(O) P R 32 (where p is O
  • Alkenyl refers to a straight or branched hydrocarbon chain radical group consisting solely of carbon and hydrogen atoms, containing at least one double bond, having from two to twelve carbon atoms, preferably two to eight carbon atoms and which is attached to the rest of the molecule by a single bond, e.g., ethenyl, prop-1-enyl, but-1-enyl, pent-1-enyl, penta-1 ,4-dienyl, and the like.
  • an alkenyl group may be optionally substituted by one of the following groups: alkyl, alkenyl, halo, haloalkenyl, cyano, nitro, aryl, cycloalkyl, heterocyclyl, heteroaryl, oxo, trimethylsilanyl, -OR 30 , -OC(O)-R 30 , -N(R 30 ) 2 , -C(O)R 30 , -C(O)OR 30 , -C(O)N(R 30 ) 2 , -N(R 30 )C(O)OR 32 , -N(R 30 )C(O)R 32 , -N(R 30 )S(O),R 32 (where t is 1 to 2), -S(O) t OR 32 (where t is 1 to 2), -S(O) P R 32 (where p is 0 to 2), and -S(O) t N(
  • Alkynyl refers to a straight or branched hydrocarbon chain radical group comprising solely of carbon and hydrogen atoms, containing at least one triple bond, optionally containing at least one double bond, having from two to twelve carbon atoms, preferably two to eight carbon atoms and which is attached to the rest of the molecule by a single bond, for example, ethynyl, propynyl, butynyl, pentynyl, hexynyl, and the like.
  • an alkynyl group may be optionally substituted by one or more of the following substituents: alkyl, alkenyl, halo, haloalkenyl, cyano, nitro, aryl, cycloalkyl, heterocyclyl, heteroaryl, oxo, trimethylsilanyl, -OR 30 , -OC(O)-R 30 , -N(R 30 ) 2 , -C(O)R 30 , -C(O)OR 30 , -C(O)N(R 30 ) 2 , -N(R 30 )C(O)OR 32 , -N(R 30 )C(O)R 32 , -N(R 30 )S(O) ( R 32 (where t is 1 to 2), -S(O) 1 OR 32 (where t is 1 to 2), -S(O) P R 32 (where p is O to 2), and -S(O)
  • Alkylene or "alkylene chain” refers to a straight or branched divalent hydrocarbon chain linking the rest of the molecule to a radical group, consisting solely of carbon and hydrogen, containing no unsaturation and having from one to twelve carbon atoms, e.g., methylene, ethylene, propylene, n-butylene, and the like.
  • the alkylene chain is attached to the rest of the molecule through a single bond and to the radical group through a single bond.
  • the points of attachment of the alkylene chain to the rest of the molecule and to the radical group can be through one carbon or any two carbons within the chain.
  • an alkylene chain may be optionally substituted by one of the following groups: alkyl, alkenyl, halo, haloalkenyl, cyano, nitro, aryl, cycloalkyl, heterocyclyl, heteroaryl, oxo, trimethylsilanyl, -OR 30 , -OC(O)-R 30 , -N(R 30 ) 2 , -C(O)R 30 , -C(O)OR 30 , -C(O)N(R 30 ) 2 , -N(R 30 )C(O)OR 32 , -N(R 30 )C(O)R 32 , -N(R 30 )S(O) t R 32 (where t is 1 to 2), -S(O) 1 OR 32 (where t is 1 to 2), -S(O) P R 32 (where p is O to 2), and -S(O),N(R 30 ,
  • alkenylene or “alkenylene chain” refers to a straight or branched divalent hydrocarbon chain linking the rest of the molecule to a radical group, consisting solely of carbon and hydrogen, containing at least one double bond and having from two to twelve carbon atoms, e.g., ethenylene, propenylene, n-butenylene, and the like.
  • the alkenylene chain is attached to the rest of the molecule through a single bond and to the radical group through a double bond or a single bond.
  • the points of attachment of the alkenylene chain to the rest of the molecule and to the radical group can be through one carbon or any two carbons within the chain.
  • an alkenylene chain may be optionally substituted by one of the following groups: alkyl, alkenyl, halo, haloalkenyl, cyano, nitro, aryl, cycloalkyl, heterocyclyl, heteroaryl, oxo, trimethylsilanyl, -OR 30 , -OC(O)-R 30 , -N(R 30 ) 2 , -C(O)R 30 , -C(O)OR 30 , -C(O)N(R 30 ) 2 , -N(R 30 )C(O)OR 32 , -N(R 30 )C(O)R 32 , -N(R 30 )S(O) t R 32 (where t is 1 to 2), -S(O) 1 OR 32 (where t is 1 to 2), -S(O) P R 32 (where p is O to 2), and -S(O) t N(
  • Alkoxy refers to a radical of the formula -OR 3 where R a is an alkyl radical as defined above containing one to twelve carbon atoms.
  • R a is an alkyl radical as defined above containing one to twelve carbon atoms.
  • the alkyl part of the alkoxy radical may be optionally substituted as defined above for an alkyl radical.
  • Alkoxyalkyl refers to a radical of the formula -R b -O-R a where R b is an alkylene chain as defined above and R 3 is an alkyl radical as defined above.
  • the oxygen atom may be bonded to any carbon in the alkylene chain and in the alkyl radical.
  • the alkyl part of the alkoxyalkyl radical may be optionally substituted as defined above for an alkyl group.
  • the alkylene chain part of the alkoxyalkyl radical may be optionally substituted as defined above for an alkylene chain.
  • Aryl refers to a hydrocarbon ring system radical comprising hydrogen, 6 to 18 carbon atoms and at least one aromatic ring.
  • the aryl radical may be a monocyclic, bicyclic, tricyclic or tetracyclic ring system, which may included fused or bridged ring systems.
  • Aryl radicals include, but are not limited to, aryl radicals derived from aceanthrylene, acenaphthylene, acephenanthrylene, anthracene, azulene, benzene, chrysene, fluoranthene, fluorene, as-indacene, s-indacene, indane, indene, naphthalene, phenalene, phenanthrene, pleiadene, pyrene, and triphenylene.
  • aryl or the prefix “ar-” (such as in “aralkyl”) is meant to include aryl radicals optionally substituted by one or more substituents independently selected from the group consisting of alkyl, alkenyl, halo, haloalkyl, haloalkenyl, cyano, nitro, aryl, aralkyi, heteroaryl, heteroarylalkyl, -R 31 -OR 30 , -R 31 -OC(O)-R 30 , -R 31 -N(R 30 ) 2 , -R 31 -C(O)R 30 , -R 31 -C(O)OR 30 , -R 31 -C(O)N(R 30 ) 2 , -R 31 -N(R 30 )C(O)OR 32 , -R 31 -N(R 30 )C(O)R 32 , -R 31 -N(R 30 )C(O)R 32 , -
  • each R 30 is independently hydrogen, alkyl, haloalkyl, cycloalkyl, cycloalkylalkyl, aryl, aralkyl, heterocyclyl, heterocyclylalkyl, heteroaryl or heteroarylalkyl; each R 31 is independently a direct bond or a straight or branched alkylene or alkenylene chain; and each R 32 is alkyl, haloalkyl, cycloalkyl, cycloalkylalkyl, aryl,
  • Alkyl refers to a radical of the formula -R b -R c where R b is an alkylene chain as defined above and R c is one or more aryl radicals as defined above, for example, benzyl, diphenylmethyl and the like.
  • the alkylene chain part of the aralkyl radical may be optionally substituted as described above for an alkylene chain.
  • the aryl part of the aralkyl radical may be optionally substituted as described above for an aryl group.
  • Alkenyl refers to a radical of the formula -R d -R c where R d is an alkenylene chain as defined above and R c is one or more aryl radicals as defined above.
  • the aryl part of the aralkenyl radical may be optionally substituted as described above for an aryl group.
  • the alkenylene chain part of the aralkenyl radical may be optionally substituted as defined above for an alkenylene group.
  • Cycloalkyl refers to a stable non-aromatic monocyclic or polycyclic hydrocarbon radical consisting solely of carbon and hydrogen atoms, which may include fused or bridged ring systems, having from three to fifteen carbon atoms, preferably having from three to ten carbon atoms, and which is saturated or unsaturated and attached to the rest of the molecule by a single bond.
  • Monocyclic radicals include, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptly, and cyclooctyl.
  • Polycyclic radicals include, for example, adamantyl, norbornyl, decalinyl, 7,7-dimethyl-bicyclo[2.2.1]heptanyl, and the like.
  • cycloalkyl is meant to include cycloalkyl radicals which are optionally substituted by one or more substituents independently selected from the group consisting of alkyl, alkenyl, halo, haloalkyl, haloalkenyl, cyano, nitro, oxo, aryl, aralkyl, cycloalkyl, cycloalkylalkyl, heterocyclyl, heterocyclylalkyl, heteroaryl, heteroarylalkyl, -R 31 -OR 30 , -R 31 -OC(O)-R 30 , -R 31 -N(R 30 ) 2 , -R 31 -C(O)R 30 , -R 31 -C(O)R 30 , -R
  • Cycloalkylalkyl refers to a radical of the formula -R b R g where R b is an alkylene chain as defined above and R 9 is a cycloalkyl radical as defined above.
  • the alkylene chain and the cycloalkyl radical may be optionally substituted as defined above.
  • “Fused” refers to any ring structure described herein which is fused to an existing ring structure in the compounds of the invention. When the fused ring is a heterocyclyl ring or a heteroaryl ring, any carbon atom on the existing ring structure which becomes part of the fused heterocyclyl ring or the fused heteroaryl ring may be replaced with a nitrogen atom.
  • Halo refers to bromo, chloro, fluoro or iodo.
  • Haloalkyl refers to an alkyl radical, as defined above, that is substituted by one or more halo radicals, as defined above, e.g., trifluoromethyl, difluoromethyl, trichloromethyl, 2,2,2-trifluoroethyl, 1-fluoromethyl-2-fluoroethyl, 3-bromo-2-fluoropropyl, i-bromomethyl-2-bromoethyl, and the like.
  • the alkyl part of the haloalkyl radical may be optionally substituted as defined above for an alkyl group.
  • Haloalkoxy refers to a radical of the formula -OR j where R j is a haloalkyl radical as defined above, e.g., trifluoromethoxy, difluoromethoxy, trichloromethoxy, 2,2,2-trifluoroethoxy, 1 -fluoromethyl-2-fluoroethoxyl, 3-bromo-2-fluoropropoxy, i-bromomethyl-2-bromoethoxy, and the like.
  • the alkyl part of the haloalkoxy radical may be optionally substituted as defined above for an alkyl group.
  • Haloalkenyl refers to an alkenyl radical, as defined above, that is substituted by one or more halo radicals, as defined above.
  • the alkenyl part of the haloalkyl radical may be optionally substituted as defined above for an alkenyl group.
  • Heterocyclyl refers to a stable 3- to 18-membered non-aromatic ring radical which consists of two to twelve carbon atoms and from one to six heteroatoms selected from the group consisting of nitrogen, oxygen and sulfur.
  • the heterocyclyl radical may be a monocyclic, bicyclic, tricyclic or tetracyclic ring system, which may include fused or bridged ring systems; and the nitrogen, carbon or sulfur atoms in the heterocyclyl radical may be optionally oxidized; the nitrogen atom may be optionally quatemized; and the heterocyclyl radical may be partially or fully saturated.
  • heterocyclyl radicals include, but are not limited to, dioxolanyl, thienyl[1 ,3]dithianyl, decahydroisoquinolyl, imidazolinyl, imidazolidinyl, isothiazolidinyl, isoxazolidinyl, morpholinyl, octahydroindolyl, octahydroisoindolyl, 2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxopyrrolidinyl, oxazolidinyl, piperidinyl, piperazinyl, 4-piperidonyl, pyrrolidinyl, pyrazolidinyi, quinuclidinyl, thiazolidinyl, tetrahydrofuryl, trithianyl, tetrahydropyranyl, thiomorpholinyl, thiamorpholinyl, 1-oxo-o-
  • heterocyclyl is meant to include heterocyclyl radicals as defined above which are optionally substituted by one or more substituents selected from the group consisting of alkyl, alkenyl, halo, haloalkyl, haloalkenyl, cyano, oxo, thioxo, nitro, aryl, aralkyl, cycloalkyl, cycloalkylalkyl, heterocyclyl, heterocyclylalkyl, heteroaryl, heteroarylalkyl, -R 31 -OR 30 , -R 31 -OC(O)-R 30 , -R 31 -N(R 30 ) 2 , -R 31 -C(O)R 30 , -R 31 -C(O)OR 30 , -R 31 -C(O)N(R 30 ) 2 , -R 31 -N(R 30 )C(O)OR 32 ,
  • ⁇ /-heterocyclyl refers to a heterocyclyl radical as defined above containing at least one nitrogen and where the point of attachment of the heterocyclyl radical to the rest of the molecule is through a nitrogen atom in the heterocyclyl radical.
  • An ⁇ /-heterocyclyl radical may be optionally substituted as described above for heterocyclyl radicals.
  • Heterocyclylalkyl refers to a radical of the formula -R b R h where R b is an alkylene chain as defined above and R h is a heterocyclyl radical as defined above, and if the heterocyclyl is a nitrogen-containing heterocyclyl, the heterocyclyl may be attached to the alkyl radical at the nitrogen atom.
  • the alkylene chain of the heterocyclylalkyl radical may be optionally substituted as defined above for an alkylene chain.
  • the heterocyclyl part of the heterocyclylalkyl radical may be optionally substituted as defined above for a heterocyclyl group.
  • Heteroaryl refers to a 5- to 14-membered ring system radical comprising hydrogen atoms, one to thirteen carbon atoms, one to six heteroatoms selected from the group consisting of nitrogen, oxygen and sulfur, and at least one aromatic ring.
  • the heteroaryl radical may be a monocyclic, bicyclic, tricyclic or tetracyclic ring system, which may include fused or bridged ring systems; and the nitrogen, carbon or sulfur atoms in the heteroaryl radical may be optionally oxidized; the nitrogen atom may be optionally quaternized.
  • Examples include, but are not limited to, azepinyl, acridinyl, benzimidazolyl, benzthiazolyl, benzindolyl, benzodioxolyl, benzofuranyl, benzooxazolyl, benzothiazolyl, benzothiadiazolyl, benzo[£>][1 ,4]dioxepinyl, 1 ,4-benzodioxanyl, benzonaphthofuranyl, benzoxazolyl, benzodioxolyl, benzodioxinyl, benzopyranyl, benzopyranonyl, benzofuranyl, benzofuranonyl, benzothienyl (benzothiophenyl), benzotriazolyl, benzo[4,6]imidazo[1 ,2-a]pyridinyl, carbazolyl, cinnolinyl, dibenzofuranyl, dibenzothi
  • heteroaryl is meant to include heteroaryl radicals as defined above which are optionally substituted by one or more substituents selected from the group consisting of alky!, alkenyl, alkoxy, halo, haloalkyl, haloalkenyl, cyano, oxo, nitro, thioxo, aryl, aralkyl, cycloalkyl, cycloalkylalkyl, heterocyclyl, heterocyclylalkyl, heteroaryl, heteroarylalkyl, -R 31 -OR 30 , -R 31 -OC(O)-R 30 , -R 31 -N(R 30 ) 2 , -R 31 -C(O)R 30 , -R 31 -C(O)OR 30 , -R 31 -C(O)N(R 30 ) 2 , -R 31 -N(R 30 )
  • each R 30 is independently hydrogen, alkyl, alkenyl, haloalkyl, cycloalkyl, cycloalkylalkyl, aryl, aralkyl, heterocyclyl, heterocyclylalkyl, heteroaryl or heteroarylalkyl; each R 31 is independently a direct bond or a straight or branched alkylene or alkenylene chain; and each R 32 is alkyl, alkenyl, haloalkyl, cycloalkyl, cycloalkylalkyl, aryl, aralkyl, heterocyclyl, heterocyclylalkyl, heteroaryl or heteroarylalkyl.
  • ⁇ /-heteroaryl refers to a heteroaryl radical as defined above containing at least one nitrogen and where the point of attachment of the heteroaryl radical to the rest of the molecule is through a nitrogen atom in the heteroaryl radical.
  • An ⁇ /-heteroaryl radical may be optionally substituted as described above for heteroaryl radicals.
  • Heteroarylalkyl refers to a radical of the formula -R b Ri where R b is an alkylene chain as defined above and Ri is a heteroaryl radical as defined above.
  • the heteroaryl part of the heteroarylalkyl radical may be optionally substituted as defined above for a heteroaryl group.
  • the alkylene chain part of the heteroarylalkyl radical may be optionally substituted as defined above for an alkylene chain.
  • “Mammal” includes humans and both domestic animals such as laboratory animals and household pets, (e.g., cats, dogs, swine, cattle, sheep, goats, horses, rabbits), and non-domestic animals such as wildlife and the like.
  • Optional or “optionally” means that the subsequently described event of circumstances may or may not occur, and that the description includes instances where said event or circumstance occurs and instances in which it does not.
  • optionally substituted aryl means that the aryl radical may or may not be substituted and that the description includes both substituted aryl radicals and aryl radicals having no substitution.
  • substitutents on the functional group are also “optionally substituted” and so on, for the purposes of this invention, such iterations are limited to five, preferably such iterations are limited to two.
  • “Pharmaceutically acceptable iron chelator” includes any molecule capable of chelating non-heme iron in a mammal and which has been approved by an appropriate regulating authority, such as the United States Food and Drug Administration, as being acceptable for use in mammals, preferably in humans.
  • “Pharmaceutically acceptable carrier, diluent or excipient” includes without limitation any adjuvant, carrier, excipient, glidant, sweetening agent, diluent, preservative, dye/colorant, flavor enhancer, surfactant, wetting agent, dispersing agent, suspending agent, stabilizer, isotonic agent, solvent, or emulsifier which has been approved by an appropriate regulating authority, such as the United States Food and Drug Administration, as being acceptable for use in humans or domestic animals.
  • “Pharmaceutically acceptable salt” includes both acid and base addition salts.
  • “Pharmaceutically acceptable acid addition salt” refers to those salts which retain the biological effectiveness and properties of the free bases, which are not biologically or otherwise undesirable, and which are formed with inorganic acids such as, but are not limited to, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid and the like, and organic acids such as, but not limited to, acetic acid, 2,2-dichloroacetic acid, adipic acid, alginic acid, ascorbic acid, aspartic acid, benzenesulfonic acid, benzoic acid, 4-acetamidobenzoic acid, camphoric acid, camphor-10-sulfonic acid, capric acid, caproic acid, caprylic acid, carbonic acid, cinnamic acid, citric acid, cyclamic acid, dodecylsulfuric acid, ethane-1 ,2-disulfonic acid, ethanesulfonic acid, 2-hydroxyethanesulf
  • “Pharmaceutically acceptable base addition salt” refers to those salts which retain the biological effectiveness and properties of the free acids, which are not biologically or otherwise undesirable. These salts are prepared from addition of an inorganic base or an organic base to the free acid. Salts derived from inorganic bases include, but are not limited to, the sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, aluminum salts and the like. Preferred inorganic salts are the ammonium, sodium, potassium, calcium, and magnesium salts.
  • Salts derived from organic bases include, but are not limited to, salts of primary, secondary, and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines and basic ion exchange resins, such as ammonia, isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, diethanolamine, ethanolamine, deanol, 2-dimethylaminoethanol, 2-diethylaminoethanol, dicyclohexylamine, lysine, arginine, histidine, caffeine, procaine, hydrabamine, choline, betaine, benethamine, benzathine, ethylenediamine, glucosamine, methylglucamine, theobromine, triethanolamine, tromethamine, purines, piperazine, piperidine, ⁇ /-ethylpiperidine, polyamine resins and the like.
  • basic ion exchange resins
  • Particularly preferred organic bases are isopropylamine, diethylamine, ethanolamine, trimethylamine, dicyclohexylamine, choline and caffeine.
  • a “pharmaceutical composition” refers to a formulation of a biologically active ingredient and a medium generally accepted in the art for the delivery of the biologically active ingredient to mammals, e.g., humans. Such a medium includes all pharmaceutically acceptable carriers, diluents or excipients therefor.
  • “Therapeutically effective amount” refers to that amount of a biologically active ingredient which, when administered to a mammal, preferably a human, is sufficient to effect treatment, as defined below, of an iron overload disorder in the mammal, preferably a human.
  • a biologically active ingredient which constitutes a “therapeutically effective amount” will vary depending on the active ingredient, the iron overload disorder and its severity, the manner of administration, and the age of the mammal to be treated, but can be determined routinely by one of ordinary skill in the art having regard to his own knowledge and to this disclosure.
  • a “therapeutically effective amount” is that amount of an active ingredient sufficient to produce a negative iron balance in the mammal treated.
  • Treating" or “treatment”, as used herein, covers the treatment of an iron overload disorder in a mammal, preferably a human and includes:
  • Active ingredient refers to a DMT1 inhibitor (either an agent directly inhibiting DMT1 or an agent that reduces the driving force for iron influx in enterocytes), a pharmaceutically acceptable iron chelator, a calcium channel blocker or a proton pump inhibitor utilized in the combination therapies of the invention.
  • the terms “disease” and “condition” may be used interchangeably or may be different in that the particular malady or condition may not have a known causative agent (so that etiology has not yet been worked out) and it is therefore not yet recognized as a disease but only as an undesirable condition or syndrome, wherein a more or less specific set of symptoms have been identified by clinicians.
  • combination therapy refers to the administration of more than one active ingredient for the treatment of an iron overload disorder. Unless the context makes it clear otherwise, “combination therapy” may include the administration of any mixture or permutation of more than one active ingredient . Unless the context makes it clear otherwise, “combination therapy” may include simultaneous or sequential administration of two or more different active ingredients, in any order, one after the other. Unless the context makes clear otherwise, “combination therapy” may include dosage forms of one active ingredient combined with the dosage forms of another active ingredient. Unless the context makes clear otherwise, “combination therapy” may include different routes of administration for each active ingredient. Dosage forms, routes of administration and pharmaceutical compositions include, but are not limited to, those described herein.
  • stereoisomer refers to a compound made up of the same atoms bonded by the same bonds but having different three-dimensional structures, which are not interchangeable.
  • the present invention contemplates various stereoisomers and mixtures thereof and includes “enantiomers”, which refers to two stereoisomers whose molecules are nonsuperimposeable mirror images of one another.
  • a “tautomer” refers to a proton shift from one atom of a molecule to another atom of the same molecule.
  • the present invention includes tautomers of any said compounds.
  • Inhibition of iron influx in enterocytes may be achieved either by direct inhibition of DMT1 or indirectly by removing the driving force for electrogenic iron influx.
  • Iron transport via DMT1 in enterocytes involves an inwardly rectifying cation influx (Xu et a/., PIoS Biology 2: 378-386, 2004).
  • cation influx requires a counter- current to maintain the driving force for cation entry.
  • Blocking this counter-current will inhibit iron influx and such block can be used in isolation or in combination with direct inhibition of DMT1 , with a chelator or with a proton pump inhibitor.
  • the counter-current is provided by potassium efflux through an ion channel selectively permeable to potassium or to chloride influx through an ion channel selectively permeable to chloride.
  • DMT1 inhibitors utilized in the combination therapies of the invention are the compounds described in PCT Published Application No. WO 2008/109840; PCT Published Application No. WO 2008/115999; PCT Published Application No. WO 2008/118790; PCT Published Application No. WO 2008/121861 ; and PCT Published Application No. WO 2008/151288.
  • DMT1 inhibitors utilized in the combination therapy of the invention are compounds of formula (I); as described in PCT Published Application No. WO 2008/109840:
  • n and m are each independently 0, 1 or 2;
  • R 1 and R 2 are each independently a direct bond, -C(R 9 ) 2 -, -S-, -O-, -C(O)-, -N(R 9 )- or R 3 and R 4 are different and are each independently selected from
  • R 5 and R 6 are different and are each independently selected from hydrogen, alkyl, halo, haloalkyl, -R 11 -CN, -R 11 -NO 2 , -R 11 -N(R 14 ) 2 , -R 11 -C(O)OR 14 ,
  • each R 7 and R 8 is independently selected from alkyl, alkenyl, alkynyl, halo, haloalkyl, haloalkenyl, haloalkoxy, optionally substituted cycloalkyl, optionally substituted cycloalkylalky
  • R 10 is -C(R 9 ) 2 -, -S-, -O- or -N(R 9 )-; each R 11 is independently a direct bond or a straight or branched alkylene chain; each R 12 and R 13 is independently hydrogen, alkyl, or -OR 9 ; each R 14 is independently hydrogen, alkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heteroaryl or optionally substituted heteroaryl; and each R 15 is alky!; as stereoisomers, enantiomers, tautomers thereof or mixtures thereof; or pharmaceutically acceptable salts thereof.
  • DMT1 inhibitors utilized in the combination therapy of the invention are compounds of formula (II), as also described in PCT Published Application No. WO 2008/109840:
  • q and r are each independently 0, 1 or 2;
  • R i18 and R ,19 are different and are each independently selected from
  • each R 24 is hydrogen, alkyl, alkenyl, alkynyl, haloalkyl, alkoxyalkyl, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heterocyclyl, optionally substituted heterocyclylalkyl, optionally substituted
  • DMT1 inhibitors utilized in the combination therapy of the invention are compounds of formula (I), as described in PCT Published Application No.
  • n and m are each independently 1 , 2, 3, 4, 5, 6 or 7; and are each independently aryl or heteroaryl;
  • R 1 is a direct bond, -O-, -S(O) P - (wherein p is 0, 1 or 2), -C(R 4 ) 2 -, -C(O)- or -N(R 4 )-; at least one R 2 and at least one R 3 is independently selected from of
  • R 2 's and R 3 's are each independently selected from alkyl, halo, haloalkyl, -R 6 -OR 7 ,-R 6 -CN, -R 6 -NO 2
  • DMT1 inhibitors utilized in the combination therapy of the invention are compounds of formula (I), as described in PCT Published Application No. WO 2008/118790:
  • R 1 is an optionally substituted aryl or an optionally substituted heteroaryl
  • R 2 is -OR 5 , -OC(O)R 5 , or -S(O) P R 5 (wherein p is 0, 1 or 2); each R 3 is independently selected from alkyl, alkenyl, alkynyl, halo, haloalkyl, haloalkenyl, haloalkynyl, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted cycloalkylalkenyl, optionally substituted cycloalkylalkynyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted aralkenyl, optionally substituted aralkynyl, optionally substituted heterocyclyl, optionally substituted heterocyclylalkyl, optionally substituted heterocyclylalkyl, optionally substituted heterocyclylalkenyl, optionally substituted heterocyclylalkynyl, optionally substituted heteroaryl
  • R 4 is -[C(R 6 ) 2 ] n - wherein n is 2 or 3;
  • R 5 is hydrogen or alkyl
  • each R 6 is independently hydrogen, alkyl, alkenyl, alkynyl, haloalkyl, alkoxyalkyl, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heterocyclyl, optionally substituted heterocyclylalkyl, optionally substituted heteroaryl or optionally substituted heteroarylalkyl; each R 7 is independently alkyl, alkenyl, alkynyl, haloalkyl, alkoxyalkyl, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heterocyclyl, optionally substituted heterocyclylalkyl, optionally substituted heteroaryl or optionally substituted heteroarylalkyl; and each R 8 is a direct
  • DMT1 inhibitors utilized in the combination therapy of the invention are compounds of formula (I), as described in PCT Published Application No.
  • R 1 is hydrogen, hydroxyalkyl, optionally substituted aryl, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted aralkyl, optionally substituted heteroaryl or optionally substituted heteroarylalkyl;
  • R 2 is hydrogen, alkyl, alkenyl, alkynyl, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted cycloalkylalkenyl, optionally substituted cycloalkylalkynyl, optionally substituted heterocyclyl, optionally substituted heterocyclylalkyl, optionally substituted heterocyclylalkenyl or optionally substituted heterocyclylalkynyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted aralkenyl, optionally substituted aralkynyl, optionally substituted heteroaryl, optionally substituted heteroarylalkyl, optionally substituted heteroarylalkenyl or optionally substituted heteroarylalkynyl; R 3 is optionally substituted aryl or optionally substituted heteroaryl;
  • R 4 is alkyl, halo, haloalkyl, -OR 5 , -OC(O)R 5 , -OS(O) 2 R 5 , -S(O) P R 5 (wherein p is O, 1 or 2), or -N(R 5 ) 2 ; and, each R 5 is independently hydrogen, alkyl or haloalkyl; as stereoisomers, enantiomers, tautomers thereof or mixtures thereof; or pharmaceutically acceptable salts thereof.
  • DMT1 inhibitors utilized in the combination therapy of the invention are compounds of formula (I), as also described in PCT Published
  • R 7 is hydrogen, hydroxyalkyl, optionally substituted aryl, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted aralkyl, optionally substituted heteroaryl or optionally substituted heteroarylalkyl;
  • R 8 is hydrogen, alkyl, alkenyl, alkynyl, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted cycloalkylalkenyl, optionally substituted cycloalkylalkynyl, optionally substituted heterocyclyl, optionally substituted heterocyclylalkyl, optionally substituted heterocyclylalkenyl or optionally substituted heterocyclylalkynyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted aralkenyl, optionally substituted aralkynyl, optionally substituted heteroaryl, optionally substituted heteroarylalkyl, optionally substituted heteroarylalkeny! or optionally substituted heteroarylalkynyl;
  • R 9 is hydrogen, optionally substituted aryl, optionally substituted aralkyl, -C(O)OR 11 or -C(O)N(R 11 ) 2 ;
  • R 10 is hydrogen, alkenyl, alkynyl, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted cycloalkylalkenyl, optionally substituted cycloalkylalkynyl, optionally substituted heterocyclyl, optionally substituted heterocyclylalkyl, optionally substituted heterocyclylalkenyl or optionally substituted heterocyclylalkynyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted aralkenyl, optionally substituted aralkynyl, optionally substituted heteroaryl, optionally substituted heteroarylalkyl, optionally substituted heteroarylalkenyl or optionally substituted heteroarylalkynyl; and each R 11 is independently hydrogen, alkyl, haloalkyl, optionally substituted aryl, or optionally substituted heteroaryl; as stereoisomers, enantiomers,
  • DMT1 inhibitors utilized in the combination therapy of the invention are compounds of formula (I), as also described in PCT Published
  • R 12 is hydrogen, hydroxyalkyl, optionally substituted aryl, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted aralkyl, optionally substituted heteroaryl or optionally substituted heteroarylalkyl;
  • R 13 is hydrogen, alkyl, alkenyl, alkynyl, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted cycloalkylalkenyl, optionally substituted cycloalkylalkynyl, optionally substituted heterocyclyl, optionally substituted heterocyclylalkyl, optionally substituted heterocyclylalkenyl or optionally substituted heterocyclylalkynyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted aralkenyl, optionally substituted aralkynyl, optionally substituted heteroarylalkyl, optionally substitute
  • R 16 is optionally substituted aryl, optionally substituted aralkyl, optionally substituted heteroaryl or optionally substituted heteroarylalkyl; as stereoisomers, enantiomers, tautomers thereof or mixtures thereof; or pharmaceutically acceptable salts thereof.
  • DMT1 inhibitors utilized in the combination therapy of the invention are compounds of formula (I), as also described in PCT Published
  • R 17 is hydrogen, hydroxyalkyl, optionally substituted aryl, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted aralkyl, optionally substituted heteroaryl or optionally substituted heteroarylalkyl;
  • R 18 is hydrogen, alkyl, alkenyl, alkynyl, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted cycloalkylalkenyl, optionally substituted cycloalkylalkynyl, optionally substituted heterocyclyl, optionally substituted heterocyclylalkyl, optionally substituted heterocyclylalkenyl or optionally substituted heterocyclylalkynyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted aralkenyl, optionally substituted aralkynyl, optionally substituted heteroaryl, optionally substituted heteroarylalkyl, optionally substituted heteroarylalkenyl or optionally substituted heteroarylalkynyl;
  • R 19 is hydrogen, alkyl, halolalkyl, -C(O)OR 21 or -C(O)N(R 21 ) 2 ;
  • R 22 is optionally substituted aryl, optionally substituted aralkyl, optionally substituted heteroaryl or optionally substituted heteroarylalkyl; as stereoisomers, enantiomers, tautomers thereof or mixtures thereof; or pharmaceutically acceptable salts thereof.
  • DMT1 inhibitors utilized in the combination therapy of the invention are compounds of formula (I), as described in PCT Published Application No.
  • m is O, 1 , 2, 3, or 4; is aryl or heteroaryl;
  • DMT1 inhibitors disclosed herein can be prepared by the methods disclosed in PCT Published Application No. WO 2008/109840; PCT Published
  • DMT1 inhibitors will meet some or all of the following criteria. Oral availability should be at less than 5%. Animal model efficacy is less than about 0.1 ⁇ g/Kg to about 100 mg/Kg body weight and the target human dose is between 0.1 ⁇ g/Kg to about 100 mg/Kg body weight, although dosages outside of this range may be acceptable ("mg/Kg” means milligrams of compound per kilogram of body mass of the subject to whom it is being administered).
  • the potency (as expressed by IC 50 value) should be less than 10 ⁇ M, preferably below 1 ⁇ M and most preferably below 50 nM.
  • the IC 50 (“Inhibitory Concentration - 50%”) is a measure of the amount of compound required to achieve 50% inhibition of DMT1 , over a specific time period, in an assay of the invention.
  • Iron chelators are small molecule ligands that bind iron as Fe 3+ with high affinity. The iron bound to the iron chelators is then excreted out of the body typically in the feces, urine or both. Iron chelators are able to induce a net iron excretion in a dose dependent way. Typically, clinically relevant iron excretion is between 0.1 mg/Kg to 0.5 mg/Kg per day.
  • iron chelators have exhibited side effects such as acute renal failure or serum creatinine increases above acceptable and normal levels.
  • the serum creatinine level increases in a dose dependent way. If these side effects are observed, it is recommended that patients receive a dose reduction, interruption or discontinuation of treatment with the iron chelator.
  • Exjade® Deferasirox, [4-[(3Z,5E)- 3,5-bis(6-oxo-1-cyclohexa-2,4-dienylidene)-1 ,2,4-triazolidin-1-yl]benzoic acid, Novartis
  • Exjade® Deferasirox, [4-[(3Z,5E)- 3,5-bis(6-oxo-1-cyclohexa-2,4-dienylidene)-1 ,2,4-triazolidin-1-yl]benzoic acid, Novartis
  • Exjade® Deferasirox, [4-[(3Z,5E)- 3,5-bis(6-oxo-1-cyclohexa-2,4-dienylidene)-1 ,2,4-triazolidin-1-yl]benzoic acid, Novartis
  • Exjade® Deferasirox, [4-[(3Z,5E)- 3,5-bis(6-oxo-1
  • chelation treatment is also recommended to be dose modified by reducing the daily dose, interrupted or discontinued.
  • the present invention contemplates reducing the incidence of these side effects of chelation therapy by using lower dosages of an iron chelator in the combination therapy.
  • the invention provides a combination therapy for treating iron overload disorders without utilizing an iron chelator in order to avoid the side effects.
  • iron chelators examples include Deferasirox, Deferoxamine (Desferal®, DFO, Desferoxamine, ⁇ /-[5- ⁇ 3-[(5- aminopentyl)hydroxycarbamoyl]propionamido ⁇ pentyl]-3- ⁇ [5-/V- hydroxyacetamido)pentyl]carbamoyl ⁇ propionohydroxamic acid or pharmaceutically acceptable salts thereof, Novartis), Deferiprone (FerriproxTM, (3-hydroxy-1 ,2-dimethyl- 4(1 H)-pyridone), Apotex), Deferitrin (GT-56-252, Genzyme), Dexrazoxane ((+)-( S)-4,4'- propylenedi-2,6-piperazinedione), Desferrithiocin (DFT), Desferri-exochelin (D-Exo
  • Still other pharmaceutically acceptable iron chelators contemplated by the present invention include those described in the following U.S. Patent Application Nos. 10/300,071 ; 10/491 ,310; 10/534,357; 10/544,570; 10/580,011 ; 10/803,724; 11/011 ,110; and 11/367,042; and U.S. Patent Nos.
  • the proton pump inhibitors (PPIs) utilized in the combination therapies of the invention are compounds having H + , K + -ATPase inhibiting activity. PPIs suppress gastric acid secretion by specific inhibition of the H + , K + -ATPase in the gastric parietal cell. This inhibition of gastric acid is thought to lead to a higher pH in the stomach and gut which could lead to a decrease in the solubilization of non-heme iron and subsequently to decrease its absorption into the body.
  • PPIs may, if desired, be in the form of free base, free acid, salt, ester, solvates (in particular hydrates), anhydrate, amide, enantiomer, isomer, tautomer, prodrug, polymorph, or any other pharmacologically suitable derivative, or the like, that is therapeutically active.
  • PPIs of the invention include, but are not limited to, Omeprazole (Prilosec, AstraZeneca), Esomeprazole (Nexium, AstraZeneca), Dontoprazole, Perprazole, Habeprazole, Ransoprazole, Pariprazole, Lansoprazole (Prevacid, TAP Pharmaceuticals), Pantoprazole (Protonix, Wyeth-Ayerst), Leminoprazole, Nepaprazole, Rabeprazole (Aciphex, Janssen Pharmaceutica), Leminoprazole, Nepaprazole and Tenatoprazole (or Benatoprazole) (Mitsubishi Pharma).
  • Other PPIs include, but are not limited to: Soraprazan (Atlana); llaprazole (II-
  • WO 97/32854 (Dainippon); BY-686 (Altana); YM-020 (Yamanouchi); GYKI-34655 (Ivax); FPL-65372 (Aventis); Pharmaprojects No. 3264 (EP 509974) (AstraZeneca); nepaprazole (To aEiyo); HN- 11203 (Nycomed Pharma); OPC-22575; pumilacidin A (BMS); saviprazole (EP 234485) (Aventis); SKand F-95601 (GSK, discontinued); Pharmaprojects No. 2522 (EP 204215) (Pfizer); S-3337 (Aventis); RS-13232A
  • Still other proton pump inhibitors contemplated by the present invention include those described in the following U.S. Patent Nos.: 4,628,098; 4,689,333; 4,786,505;
  • PPIs as well as their salts, hydrates, esters, amides, enantiomers, isomers, tautomers, polymorphs, prodrugs, and derivatives may be prepared using standard procedures known to those skilled in the art of synthetic organic chemistry. See, e.g.,
  • Calcium channel blockers utilized in the combination therapies of the invention are compounds having the ability to interact with and block calcium transport through L-type calcium channels located on skeletal and smooth muscles. L-type calcium channel blockers also exert a direct effect on DMT1 , prolonging the activity of DMT1 , thereby stimulating renal iron excretion and decreasing serum iron levels.
  • Examples of calcium channel blockers that, if present, may be utilized in the combination therapies of the invention are selected from the group consisting of dihydropyridine compounds such as nifedipine, nicardipine, niguldipine and nimopidine. Still other calcium channel blockers contemplated by the present invention include those described in the following U.S. Patent Nos.: 7,485,653; 7,235,570; 7,098,211 ; 6,852,742; 5,143,915; 5,068,337; 5,021 ,436; 4,849,436; 4,841 ,054; and 4,833,150.
  • one embodiment of the invention is where the pharmaceutically acceptable iron chelator is selected from Deferasirox, Deferoxamine, DFO, Desferoxamine, Deferiprone, Deferitrin, Dexrazoxane, Desferrithiocin, Desferri- exochelin, C94, tachpyridine, aroylhydrazones, thiosemicarbazones, pyridoxal isonicotinyl hydrazones, ⁇ /, ⁇ /'-bis(2-hydroxybenzoyl)-ethylenediamine- ⁇ /, ⁇ /'-diacetic acid, or 1-substituted-2-alkyl-3-hydroxy-4-pyridones (including 1-(2'-carboxyethyl)-2- methyl-3-hydroxy-4-pyridone).
  • the pharmaceutically acceptable iron chelator is selected from Deferasirox, Deferoxamine, DFO, Desferoxamine, Deferiprone, Deferitrin, De
  • the pharmaceutically acceptable iron chelator is selected from Deferasirox, Deferiprone, Deferitrin or Deferoxamine.
  • the proton pump inhibitor is selected from Omeprazole, Esomeprazole, Dontoprazole, Perprazole, Habeprazole, Ransoprazole, Pariprazole, Lansoprazole, Pantoprazole, Leminoprazole, Nepaprazole, Rabeprazole, Leminoprazole, Nepaprazole, Tenatoprazole, Soraprazan, llaprazole, AZD-0865, YH-1885, ,4-(3,4-dihydro-1-methyl-2(1 W)-isoquinolinyl)- ⁇ /-(4- fluorophenyl)-5,6-dimethyl-2-pyrimidinamine monohydrochloride, BY-112, imidazol(1 ,2- a
  • proton pump inhibitor is selected from the group consisting of Omeprazole, Esomeprazole, Lansoprazole, Pantoprazole, Rabeprazole and Tenatoprazole.
  • proton pump inhibitor is selected from the group consisting of Omeprazole and Lansoprazole.
  • the calcium channel blocker is selected from the group consisting of: nifedipine, nicardipine, niguldipine and nimopidine. Another embodiment of the invention is where the calcium channel blocker is nifedipine.
  • DMT1 inhibitor is selected from:
  • DMT1 inhibitor is (2,4,6- trimethyl-1 ,3-phenylene)bis(methylene) dicarbamimidothioate dihydrochloride.
  • Another embodiment of the invention is wherein the therapeutically effective amount of the DMT1 inhibitor is a daily dosage amount from about 0.001 mg/Kg body weight to about 100 mg/Kg body weight of the mammal.
  • Another embodiment of the invention is wherein daily dosage amount of the DMT1 inhibitor is selected from about 10 mg/Kg body weight, about 25 mg/Kg body weight, or about 50 mg/Kg body weight of the mammal.
  • Another embodiment of the invention is wherein the daily dosage amount of the pharmaceutically acceptable iron chelator is from about 10 mg/Kg body weight to about
  • the daily dosage amount of the pharmaceutically acceptable iron chelator is selected from about 10 mg/Kg body weight, about 20 mg/Kg body weight, about 30 mg/Kg body weight, about 50 mg/Kg body weight, about 75 mg/Kg body weight or about 100 mg/Kg body weight of the mammal.
  • the therapeutically effective amount of the proton pump inhibitor is a daily dosage amount from about 10 mg/Kg body weight to about 100 mg/Kg body weight of the mammal.
  • the daily dosage amount of the proton pump inhibitor is selected from about 15 mg/Kg body weight, about 40 mg/Kg body weight, or about 75 mg/Kg body weight of the mammal.
  • the therapeutically effective amount of the calcium channel blocker is a daily dosage amount from about 0.001 mg/Kg body weight to about 100 mg/Kg body weight of the mammal.
  • Another embodiment of the invention is wherein the daily dosage amount of the calcium channel blocker is selected from about 0.1 mg/Kg body weight, about 2.0 mg/Kg body weight, or about 20 mg/Kg body weight of the mammal.
  • Another embodiment of the invention is wherein the therapeutically effective amount of the DMT1 inhibitor is a daily dosage amount from about 0.001 mg/Kg body weight to about 100 mg/Kg body weight of the mammal and the therapeutically effective amount of the proton pump inhibitor is a daily dosage amount from about 10 mg/Kg body weight to about 100 mg/Kg body weight of the mammal.
  • Another embodiment of the invention is wherein the daily dosage amount of the DMT1 inhibitor is selected from the group consisting of 10 mg/Kg body weight, 25 mg/Kg body weight, and 50 mg/Kg body weight of the mammal, and the daily dosage amount of the proton pump inhibitor is selected from the group consisting of 15 mg/Kg body weight, 40 mg/Kg body weight, and 75 mg/Kg body weight of the mammal.
  • DMT1 inhibitor is 25 mg/Kg body weight of the mammal and the daily dosage amount of the proton pump inhibitor is 40 mg/Kg body weight of the mammal.
  • Another embodiment is wherein the daily dosage amount of the DMT1 inhibitor is 50 mg/Kg body weight of the mammal and the daily dosage amount of the proton pump inhibitor is 75 mg/Kg body weight of the mammal.
  • Another embodiment of the invention is wherein all of the active ingredients are concomitantly administered to the mammal.
  • Another embodiment of the invention is wherein all of the active ingredients are sequentially administered to the mammal. Another embodiment of the invention is wherein at least two of the active ingredients are concomitantly administered to the mammal.
  • Another embodiment of the invention is wherein at least two of the active ingredients are sequentially administered to the mammal.
  • Another embodiment is wherein the DMT1 inhibitor and the proton pump inhibitor are concomitantly administered to the mammal.
  • Another embodiment of the invention is wherein the DMT1 inhibitor and the proton pump inhibitor are sequentially administered to the mammal.
  • the iron overload disorder is a primary iron overload disorder.
  • the primary iron overload disorder is independently selected from hereditary hemochromatosis, juvenile hemochromatosis, ferroportin disease, neonatal hemochromatosis, Bantu siderosis, African iron overload, gracile syndrome, or Friedreich Ataxia.
  • Another embodiment of the invention is where the primary iron overload is hereditary hemochromatosis.
  • Another embodiment of the invention is where the iron overload disorder is a secondary iron overload disorder.
  • the iron overload disorder is transfusional iron overload disorder.
  • the iron overload disorder is associated with a disease and/or condition independently selected from thalassemia (beta and alpha, major, minor and intermedia), hypochromic microcytic anemia, sickle cell anemia, microcytic iron loading anemia, hereditary sideroblastic anemia, congenital dyserythropoeitic anemia, porphyria cutanea tarda, pyruvate kinase deficiency, hereditary atransferrinemia, ceruloplasmin deficiency, myelodysplastic syndromes, pulmonary hemosiderosis, aceruloplasminemia or x-linked sideroblastic anemia.
  • iron overload disorder is associated with a disease and/or condition independently selected from neurodegenerative disease (including ALS, prion diseases, Parkinson's, and Alzheimers), cardiovascular disease (including atherosclerosis, ischemic cerebrovascular disease and ischemic stroke), inflammation (including arthritis and disease progression in viral hepatitis), cancer, insulin resistance, non-alcoholic liver disease, alcoholic liver disease, or infectious disease (including HIV, malaria and Yersinia infections).
  • a disease and/or condition independently selected from neurodegenerative disease (including ALS, prion diseases, Parkinson's, and Alzheimers), cardiovascular disease (including atherosclerosis, ischemic cerebrovascular disease and ischemic stroke), inflammation (including arthritis and disease progression in viral hepatitis), cancer, insulin resistance, non-alcoholic liver disease, alcoholic liver disease, or infectious disease (including HIV, malaria and Yersinia infections).
  • the present invention is directed to the use of certain DMT1 inhibitors in combination with pharmaceutically acceptable iron chelators, calcium channel blockers and/or proton pump inhibitors in treating iron overload disorders in mammals, preferably in human.
  • iron overload disorder refers to a condition in a mammal, preferably a human, wherein the level of iron in the body is elevated compared to the normal level of iron for the particular mammal, such as an elevated iron serum level compared to the normal iron serum level for the mammal or an increased level of iron in the liver of the mammal as compared to the normal level of iron in the liver in the mammal.
  • Elevated serum iron levels can be determined by direct measurement of serum iron using a colorimetric assay, or by the standard transferrin saturation assay (which reveals how much iron is bound to the protein that carries iron in the blood), or by the standard serum ferritin assay.
  • transferrin saturation levels of 45% or higher are usually indicative of abnormally high levels of iron in the serum.
  • Elevated iron levels in the liver can be determined measuring the iron content of the liver from tissue obtained by a liver biopsy or by imaging technique such as MRI and/or SQUID. The degree of iron levels in other tissues (e.g., brain, heart) may also be estimated using these and other imaging techniques.
  • the iron overload disorder is a primary iron overload disorder (including, but not limited to, hereditary hemochromatosis, juvenile hemochromatosis, ferroportin disease, neonatal hemochromatosis, Bantu siderosis, African iron overload, gracile syndrome, and Friedreich Ataxia, as well as all of the anemias listed below in which patients may not be transfused but may become iron overloaded due to increased erythroid drive and the resulting increased iron absorption in the gut) and secondary (or transfusional) iron overload disorder which can be caused by repeated transfusions used to treat a number of distinct anemias, including, but not limited to, thalassemia (beta and alpha, major, minor and intermedia), hypochromic microcytic anemias, sickle cell anemia, microcytic iron loading anemias, hereditary sideroblastic anemias, congenital dyserythropoeitic anemias, porphyria cutanea tarda, pyruvate kin
  • Divalent metal transporter-1 also known as natural resistance- associated macrophage protein-2 (NRAMP2) and divalent cation transporter-1 (DCT1 ), is an expressed transmembrane protein involved in the maintenance of iron levels in the body.
  • DMT1 is particularly important for iron absorption in the duodenum of the small intestine, where it is localized in the cytoplasm and brush border membrane of the villus enterocytes and mediates the influx of dietary non-heme iron from the intestinal lumen into the enterocytes (Gunshin et al., J. CHn. Invest, 2005, 115:1258- 1266). Once dietary iron is absorbed across the intestinal wall, there is no known physiologic mechanism for excreting iron from the body.
  • NRAMP2 natural resistance- associated macrophage protein-2
  • DCT1 divalent cation transporter-1
  • DMT1 is a primary focal point of controlling intestinal iron absorption for the maintenance of body iron homeostatsis.
  • upregulation i.e., increased activity
  • Hereditary hemochromatosis is an iron overload disorder due to intestinal iron hyperabsorption.
  • Hereditary hemochromatosis is characterized by a slow accumulation of iron from the diet to toxic levels resulting in tissue injury and multi-organ malfunction.
  • HFE hemochromatosis gene
  • DMT1 activity has also been implicated in the etiology and pathophysiology of hypochromic microcytic anemias, thalassemia, microcytic iron loading anemias, hereditary sideroblastic anemias, hereditary hypochromic anemias, congenital dyserythropoietic anemias, pyruvate kinase deficiency, hereditary atransferrinemia, and certain myelodysplastic syndromes, as there is a direct correlation between the degree of iron limited anemia, increased DMT1 expression in the duodenum and, by extension, increased iron absorption via DMT1 (Morgan et al., Blood Cells Molecules and Diseases, 2002, 29:384-399).
  • DMT1 has a role in iron overload disorders such as acquired iron overload.
  • the risk factors for acquired iron overload might include for example excessive ingestion of red meat, iron supplements or foods that are iron fortified.
  • Acquired iron overload can also occur from the use of iron cookware, drinking unpurified tap water, use of oral contraceptives, blood transfusions and cigarette smoking.
  • DMT1 pattern of expression and function supports it as a candidate target for the treatment of acquired iron overload and other related maladies.
  • DMT1 In addition to the small intestine, DMT1 is also highly expressed in the kidney suggesting a role in renal iron handling and possibly reabsorption of filtered iron (Ferguson et al., Am. J. Physiol. Renal. Physiol., 2001 , 280: F803-F814) and is also involved in the delivery of iron to peripheral tissues by transferrin (Fleming et al., Proc. Natl. Acad. Sci., 1998, 85:1148-1153).
  • DMT1 inhibitors when dosed in a fashion that increases their systemic exposure, may be useful in an acute unloading of iron via the urine, by inhibiting DMT1 expressed in the kidney
  • DMT1 may not play a crucial role in regulating urinary iron excretion.
  • studies performed in the Belgrade rat which carries a missense neomorphic mutation in DMT1 , G185R, demonstrated that the resultant loss of iron transporting activity in DMT1 had no effect on urinary iron excretion (Ferguson et al., Kidney International (2003) 64:1755-1764).
  • DMT1 may also play a role in regulating iron flux to the brain.
  • DMT1 inhibitors may act to reduce the amount of iron absorbed by the brain, when dosed in a fashion that increases their systemic exposure and allows them to play a role at the blood brain barrier or within the brain (Lehmann et al., 2006, J. Med. Genet, 2006, 43(10):e52; Schenck et al., Top. Magn Reson. Imaging., 2006,17(1 ):41-50).
  • mice that are defective in DMT1 activity develop hyprochromic microcytic anemia, a severe form of iron deficiency anemia, due to a defect in intestinal iron absorption.
  • the hfe ';' knockout mouse model of hereditary hemochromatosis is characterized by an enhanced intestinal iron uptake and total body iron overload.
  • the hfe'imk/mk double mutant mouse which carries mutations in both the HFE and DMT1 genes, fails to load iron, indicating that hemochromatosis ⁇ hfe ';' ) can be prevented by blocking the flux of iron through the DMT1 protein (Levy et al., J. Clin.
  • DMT1 is inappropriately upregulated at the intestinal brush border. This aberrant excessive expression of DMT1 in hereditary hemochromatosis is fundamental to the primary pathophysiology of this condition (Zoller et al., Gastroenterology, 2001 , 120:1412-1419).
  • DMT1 inhibitors of the invention are useful in treating iron disorders by directly interacting with a region of the DMT1 protein that modulates or controls iron flux.
  • a direct interaction is supported by electrophysiology studies and the fact that the DMT1 inhibitors of the invention are not potent inhibitors of cation flux in the closely related transporter Natural Resistance-Associated Macrophage Protein-1 (NRAMP1 ).
  • the DMT1 inhibitors of the invention modulate the activity of DMT1 downwards, thereby inhibiting the ability of DMT1 to uptake non-heme iron across the cellular membrane.
  • the combination therapies of the invention are therefore useful in treating iron overload disorders which can be ameliorated by the inhibition of DMT1 activity.
  • the combination therapies of the invention are also useful in treating or preventing symptoms, diseases and/or conditions in a mammal associated with hereditary hemochromatosis due to accumulation of iron in body tissues such as arthritis, liver disease, heart disease, impotence, early menopause, abnormal skin pigmentation, thyroid deficiency, damage to pancreas, diabetes, and damage to adrenal gland (Sheth et al., Annu. Rev. Med., 2000, 51 :443-464).
  • the combination therapies of the invention are also useful in treating other forms of hemochromatosis including, but are not limited to, juvenile hemochromatosis and neonatal hemochromatosis.
  • Juvenile hemochromatosis has a much earlier onset and exhibits more severe symptoms such as endocrine dysfunction, joint disease, and cardiac abnormalities due to excessive iron deposition from an early age.
  • Neonatal hemochromatosis is a rare fetal gestational condition that results in iron accumulation in the liver of the fetus.
  • the combination therapies of the invention are also useful in treating or preventing transfusional iron overload.
  • Chronic blood transfusion is the established therapy for thalassaemia major, bone marrow failure and complications of sickle cell anaemia and other related disorders. With hypertransfusion, the systemic iron load accumulates. Because there is no natural way for the body to eliminate the iron, the excess iron in the transfused blood builds up to cause iron overload and becomes toxic to tissues and organs, particularly the liver, heart, and pancreas. Transfusional iron overload typically results in the patient's premature death from organ failure.
  • the transfusional iron overload is unfortunately augmented by increased iron absorption, which is the natural attempt of the body to increase iron levels in order to promote erythropoiesis, which is itself compromised by the disease states above. Decreased absorption of iron by the inhibition of DMT1 activity may reduce the iron overload related to the transfusional iron overload and supports the use of DMT1 inhibitors in the combination therapies of the invention.
  • the combination therapies of the invention may be useful in reducing the daily dosage of chelation therapy and preventing the chelation therapy associated side effects in patients including, but not limited to, those with iron overload and renal impairment, and without a corresponding reduction in daily net excreted iron.
  • the combination therapies of the invention may also be useful in delaying when younger patients are required to begin chelation therapy and thereby protecting them from chelation associated toxicities by blocking the uptake of iron from the gut.
  • the combination therapies of the invention may be useful in reducing the daily dosage of PPIs and preventing the PPI associated side effects in patients, particularly those on long term use, including, but not limited to, those with iron overload and renal impairment, and without a corresponding reduction in daily net excreted iron.
  • combination therapies of the present invention will exert greater than additive effects or synergistic effects as the mechanisms of action employed for the active ingredients are different and each will probably act independently of one another.
  • Each active ingredient will act upon a different pool of iron.
  • PPIs may limit gut uptake by reducing non-heme iron solubility in the gut
  • DMT1 inhibitors will block the uptake of iron in the gut through this transporter
  • calcium channel blockers will enhance renal urinary iron excretion and iron chelators can bind and excrete iron that is already within the body.
  • the combination therapies of the invention may reduce the number or frequency of phlebotomies or therapeutic erythrocytapheresis treatments required to meet hemochromatosis patient serum iron and tissue iron therapeutic targets.
  • the combination therapies of the invention may also prevent iron overload in patients or may prevent re-loading of iron overlaod in patients that have concluded their phlebotomy treatments.
  • the general value of the combination therapies of the invention in decreasing serum iron levels can be determined using the assays described herein or below in the Biological Assays section.
  • the general value of the combination therapies of the invention in treating iron overload disorders in mammals, preferably humans may be established in industry standard animal models for demonstrating the efficacy of combination therapies in treating iron overload disorders.
  • an iron deficient anemia and therefore a hyperabsorptive iron state
  • a hyperabsorptive iron state may be induced by dietary means, or by treatment with phenylhydrazine, or by phlebotomy (Refino et al., Am. J. Clin. Nutr. 1983, 37:904-909; Redondo et al., Lab. Animal Sci. 1995, 45:578-583; Frazer ef a/., Gastroenterology, 2002, 123:835-844).
  • iron absorption can also be stimulated by creating an hypoxic state to stimulate erythropoiesis (Raja et al., Br. J. Haematol., 1988, 68:373-378).
  • combination therapies of the invention can be assessed by measuring reduced iron flux via the duodenum acutely or by monitoring whether chronic exposure to a combination therapy causes a decrease in the amount of iron loading as measured by serum iron, transferrin saturation, ferritin and liver iron.
  • iron flux in these animals can be measured by tracing the absorption of radioactive iron administered orally.
  • genetic rat models of iron overload offer another format to show efficacy of the combination therapies of the invention.
  • These models are applicable to a variety of iron disorders such as hereditary hemochromatosis (Levy et al., Blood, 1999, 94:9-11 ), juvenile hemochromatosis (Huang et al., J. Clin. Invest, 2005 115:2187-2191), beta-2-microglobulin (de Sousa et al., Immun. Lett, 1994, 39:105- 111 ), thalassemia (Ciavatta et al., Proc. Nat. Acad.
  • a combination therapy's efficacy can be assessed by measuring reduced iron flux via the duodenum acutely or by monitoring whether chronic exposure to a compound causes a decrease in the amount of iron loading as judged by serum iron, transferrin saturation, ferritin and liver iron. Alternatively, iron flux in these animals can be measured by tracing the absorption of radioactive iron administered orally.
  • a successful active ingredient of the present invention will meet some or all of the following criteria.
  • Oral availability should be at less than 5%.
  • Animal model efficacy is less than about 0.1 ⁇ g to about 100 mg/Kg body weight and the target human dose is between 0.1 ⁇ g to about 100 mg/Kg body weight, although doses outside of this range may be acceptable ("mg/Kg” means milligrams of compound per kilogram of body mass of the subject to whom it is being administered).
  • the potency (as expressed by IC 50 value) should be less than 10 ⁇ M, preferably below 1 ⁇ M and most preferably below 50 nM.
  • the IC 50 is a measure of the amount of compound required to achieve 50% inhibition of DMT1 , over a specific time period, in an assay of the invention. ADMINISTRATION OF THE COMBINATION THERAPY OF THE INVENTION
  • the recipients of the combination therapy of the invention can be any vertebrate animal, such as mammals.
  • the preferred recipients are mammals of the Orders Primate (including humans, apes and monkeys), Arteriodactyla (including horses, goats, cows, sheep, pigs), Rodenta (including mice, rats, rabbits, and hamsters), and Camivora (including cats, and dogs).
  • the preferred recipients are turkeys, chickens and other members of the same order. The most preferred recipients are humans.
  • Each active ingredient, or a pharmaceutical composition comprising a therapeutically effective amount of the active ingredient may be concomitantly administered to a mammal in need thereof.
  • concomitantly administered means the administration of the active ingredients substantially concurrently.
  • concomitantly administered encompasses not only administering the active ingredients in a single pharmaceutical dosage form but also the administration of each active ingredient in its own pharmaceutical dosage form, such as a pharmaceutical composition for oral administration.
  • each active ingredient, or a pharmaceutical composition comprising a therapeutically effective amount of the active ingredient may be sequentially administered to a mammal in need thereof.
  • sequentialially administered means the administration of the active ingredients at separately staggered times.
  • the active ingredient can be sequentially administered such that the beneficial pharmaceutical effects of the active ingredients are realized by the mammal at substantially the same time, depending on the pharmaceutically active half-life of each active ingredient.
  • the active ingredients of the combination therapy of the invention are administered in a therapeutically effective amount, which will vary depending upon a variety of factors including the activity of the specific active ingredient employed; the metabolic stability and length of action of the active ingredient; the age, body weight, general health, sex, and diet of the patient; the mode and time of administration; the rate of excretion; the drug combination; the severity of the particular disorder or condition; and the subject undergoing therapy.
  • the preferred daily dosage amount of each active ingredient will be tailored to the individual subject, as is understood and determinable by one skilled in the relevant arts, (see, e.g., Physicians' Desk Reference (latest edition); Berkowet al., eds., The Merck Manual, 16 th edition, Merck and Co., Rahway, N.J., 1992; Goodman et al., eds., Goodman and Gilman's The Pharmacological Basis of Therapeutics, 11 th edition, The MacGraw-Hill Companies, (2005); Avery's Drug Treatment: Principles and Practice of Clinical Pharmacology and Therapeutics, 3rd edition, ADIS Press, LTD., Williams and Wilkins, Baltimore, MD.
  • treatment with a combination therapy of the invention may be initiated using a daily dosage amount of each active ingredient which is less than the optimum daily dosage amount of the active ingredient. Thereafter, the dosage may be increased by small increments until the optimum effect under the circumstances is reached, i.e., until a negative iron balance in the mammal undergoing treatment.
  • a therapeutically effective daily dosage amount of a DMT1 inhibitor of the combination therapies of the invention is from about 0.001 mg/Kg body weight to about 100 mg/Kg body weight; preferably a therapeutically effective dose is from about 0.01 mg/Kg body weight to about 75 mg/Kg body weight; more preferably a therapeutically effective dose is from about 1 mg/Kg body weight to about 50 mg/Kg body weight.
  • a therapeutically effective daily dosage amount of a DMT1 inhibitor is daily dosage amount is 10 mg/Kg body weight, 25 mg/Kg body weight, or 50 mg/Kg body weight.
  • the DMT1 inhibitor may be dosed orally once, twice, three times or four or more times a day.
  • a therapeutically effective daily dosage amount of a pharmaceutically acceptable iron chelator of the combination therapies of the invention is from about 10 mg/Kg body weight to about 100 mg/Kg body weight.
  • a therapeutically effective daily dosage amount of a pharmaceutically acceptable iron chelator of the combination therapies of the invention is about 10 mg/Kg body weight , about 20 mg/Kg body weight , about 30 mg/Kg body weight , about 50 mg/Kg body weight , about 75 mg/Kg body weight or about 100 mg/Kg body weight.
  • the pharmaceutically acceptable iron chelator can be orally administered one, two, three or more times daily depending on the daily dosage. Alternatively, pharmaceutically acceptable iron chelators can be intravenously infused daily for up to 12 hours.
  • a therapeutically effective daily dosage amount of a PPI of the combination therapies of the invention is from about 1 mg to about 120 mg.
  • a therapeutically effective daily dosage of a PPI of the combination therapies of the invention is about 1 mg/Kg body weight, about 5 mg/Kg body weight, about 10 mg/Kg body weight, about 15 mg/Kg body weight, about 20 mg/Kg body weight, about 30 mg/Kg body weight, about 40 mg/Kg body weight, about 60 mg/Kg body weight, about 90 mg/Kg body weight, about 100 mg/Kg body weight, or about 120 mg/Kg body weight.
  • the daily dosage amount per body weight is 15 mg/Kg, 40 mg/Kg or 75 mg/Kg.
  • the daily dosage amount of a PPI in a combination therapy of the invention can be less than what the current recommended daily dosage is for the PPI.
  • PPIs can be administered orally once, twice or three times a day depending on the dosage.
  • Esomeprazol (Nexium) can be administered orally once a day to adults at 20 mg or 40 mg dosage, or at 10 mg or 20 mg dosage to children 1 to 11 years, or 20 mg or 40 mg dosage to children 12 to 17 years.
  • a therapeutically effective daily dosage amount of a calcium channel blocker of the combination therapies of the invention is from about 0.001 mg/Kg body weight to about 100 mg/Kg body weight; preferably a therapeutically effective dose is from about 0.01 mg/Kg body weight to about 75 mg/Kg body weight; more preferably a therapeutically effective dose is from about 0.1 mg/Kg body weight to about 25 mg/Kg body weight.
  • a therapeutically effective daily dosage amount of a calcium channel blocker is about 0.1 mg/Kg body weight, about 1.0 mg/Kg body weight, or about 10 mg/Kg body weight.
  • a therapeutically effective daily dosage amount of a calcium channel blocker is about 0.1 mg/Kg body weight, about 2.0 mg/Kg body weight, or about 20 mg/Kg body weight.
  • the daily dosage amount of a calcium channel blocker in a combination therapy of the invention can be less that what the current recommended daily dosage is for a particular calcium channel blocker.
  • the calcium channel blocker may be dosed orally once, twice, three times or four or more times a day.
  • the recommended starting dosage for immediate-release nifedipine capsules is 10 mg, taken three times daily and the recommended starting dosage for extended-release nifedipine is 30 to 60 mg, taken once daily. .
  • each active ingredient of the combination therapies of the invention may be dosed as if it is the only therapy being taken alone as each active ingredient may work additively through different mechanisms.
  • Combination therapy may be from 1 month to 12 months or from 1 year to 5 years or for the life of the recipient.
  • the combination therapies of the invention may be administered to the mammal, preferably the human, in combination with phlebotomy treatment.
  • Therapeutic phlebotomy entails periodic removal of fixed amounts of blood. Typically, phlebotomy will remove about 250 mg of iron with each treatment per week.
  • the combination therapies maybe administered prior to the initiation of phlebotomy, during phlebotomy therapy, after phlebotomy therapy has concluded or at all of these timepoints.
  • compositions comprising a therapeutically effective amount of a DMT1 inhibitor and a pharmaceutically acceptable excipient are disclosed in PCT Published Application No. WO 2008/109840; PCT Published Application No. WO 2008/115999; PCT Published Application No. WO 2008/118790; PCT Published Application No. WO 2008/121861 ; and PCT Published Application No. WO 2008/151288; the disclosures of which are incorporated in full by reference herein.
  • Pharmaceutical compositions for pharmaceutically acceptable iron chelators, proton pump inhibitors and calcium channel blockers are well-known in the art.
  • the present invention also relates to pharmaceutical compositions (hereinafter referred to as "combination-therapy pharmaceutical compositions”) comprising a therapeutically effective amount of a DMT1 inhibitor and a therapeutically effective amount of a pharmaceutically acceptable iron chelator and/or a therapeutically effective amount of a proton pump inhibitor, and/or a therapeutically effective amount of a calcium channel blocker, and a pharmaceutically acceptable excipient.
  • a pharmaceutically acceptable iron chelator and/or a therapeutically effective amount of a proton pump inhibitor
  • a calcium channel blocker a pharmaceutically acceptable excipient
  • Such combination-therapy pharmaceutical compositions can be prepared by combining the active ingredients with an appropriate pharmaceutically acceptable carrier, diluent or excipient, and may be formulated into preparations in solid, semi-solid, liquid or gaseous forms, such as tablets, capsules, powders, granules, ointments, solutions, suppositories, injections, inhalants, gels, microspheres, and aerosols.
  • Typical routes of administering such combination-therapy pharmaceutical compositions include, without limitation, oral, topical, transdermal, inhalation, parenteral, sublingual, rectal, vaginal, and intranasal.
  • parenteral as used herein includes subcutaneous injections, intravenous, intramuscular, intrasternal injection or infusion techniques.
  • Such combination-therapy pharmaceutical compositions are formulated so as to allow the active ingredients contained therein to be bioavailable upon administration of the composition to a patient.
  • the combination-therapy pharmaceutical compositions disclosed herein also contain a pharmaceutically acceptable carrier, including any suitable diluent or excipient, which includes any pharmaceutical agent that does not itself induce the production of antibodies harmful to the individual receiving the composition, and which may be administered without undue toxicity.
  • Pharmaceutically acceptable carriers include, but are not limited to, liquids, such as water, saline, glycerol and ethanol, and the like.
  • the combination-therapy pharmaceutical compositions disclosed herein may be in the form of a solid or liquid.
  • the carrier(s) are particulate, so that the compositions are, for example, in tablet or powder form.
  • the carrier(s) may be liquid, with the compositions being, for example, an oral syrup, injectable liquid or an aerosol, which is useful in, for example, inhalatory administration.
  • the combination-therapy pharmaceutical compositions disclosed herein is preferably in either solid or liquid form, where semi-solid, semi-liquid, suspension and gel forms are included within the forms considered herein as either solid or liquid.
  • the combination-therapy pharmaceutical compositions disclosed herein may be formulated into a powder, granule, compressed tablet, pill, capsule, chewing gum, wafer or the like form.
  • a solid composition will typically contain one or more inert diluents or edible carriers.
  • binders such as carboxymethylcellulose, ethyl cellulose, microcrystalline cellulose, gum tragacanth or gelatin; excipients such as starch, lactose or dextrins, disintegrating agents such as alginic acid, sodium alginate, Primogel, corn starch and the like; lubricants such as magnesium stearate or Sterotex; glidants such as colloidal silicon dioxide; sweetening agents such as sucrose or saccharin; a flavoring agent such as peppermint, methyl salicylate or orange flavoring; and a coloring agent.
  • excipients such as starch, lactose or dextrins, disintegrating agents such as alginic acid, sodium alginate, Primogel, corn starch and the like
  • lubricants such as magnesium stearate or Sterotex
  • glidants such as colloidal silicon dioxide
  • sweetening agents such as sucrose or saccharin
  • a flavoring agent such as peppermint, methyl sal
  • combination-therapy pharmaceutical compositions disclosed herein When the combination-therapy pharmaceutical compositions disclosed herein are in the form of capsules, for example, a gelatin capsule, they may contain, in addition to materials of the above type, a liquid carrier such as polyethylene glycol or oil.
  • the combination-therapy pharmaceutical compositions disclosed herein may be in the form of a liquid, for example, an elixir, syrup, solution, emulsion or suspension.
  • the liquid may be for oral administration or for delivery by injection, as two examples.
  • preferred composition contain, in addition to the active ingredients, one or more of a sweetening agent, preservatives, dye/colorant and flavor enhancer.
  • a surfactant, preservative, wetting agent, dispersing agent, suspending agent, buffer, stabilizer and isotonic agent may be included.
  • Liquid combination-therapy pharmaceutical compositions may include one or more of the following adjuvants: sterile diluents such as water for injection, saline solution, preferably physiological saline, Ringer's solution, isotonic sodium chloride, fixed oils such as synthetic mono or diglycerides which may serve as the solvent or suspending medium, polyethylene glycols, glycerin, propylene glycol or other solvents; antibacterial agents such as benzyl alcohol or methyl paraben; antioxidants such as ascorbic acid or sodium bisulfite; chelating agents such as ethylenediaminetetraacetic acid; buffers such as acetates, citrates or phosphates and agents for the adjustment of tonicity such as sodium chloride or dextrose.
  • the parenteral preparation can be enclosed in ampoules, disposable syringes or multiple dose vials made of glass or plastic.
  • Physiological saline is a preferred adjuvant
  • a liquid combination-therapy pharmaceutical composition of the invention intended for either parenteral or oral administration should contain an amount of a compound of the invention such that a suitable dosage will be obtained. Typically, this amount is at least 0.01 % of a compound of the invention in the composition. When intended for oral administration, this amount may be varied to be between 0.1 and about 70% of the weight of the composition.
  • Preferred oral combination-therapy pharmaceutical compositions contain between about 4% and about 50% of the compound of the invention.
  • Preferred combination-therapy pharmaceutical compositions and preparations according to the present invention are prepared so that a parenteral dosage unit contains between 0.01 to 10% by weight of the compound prior to dilution of the invention.
  • the combination-therapy pharmaceutical composition of the invention may be intended for topical administration, in which case the carrier may suitably comprise a solution, emulsion, ointment or gel base.
  • the base for example, may comprise one or more of the following: petrolatum, lanolin, polyethylene glycols, bee wax, mineral oil, diluents such as water and alcohol, and emulsifiers and stabilizers.
  • Thickening agents may be present in a pharmaceutical composition for topical administration.
  • the composition may include a transdermal patch or iontophoresis device.
  • Topical formulations may contain a concentration of the compound of the invention from about 0.1 to about 10% w/v (weight per unit volume).
  • the combination-therapy pharmaceutical composition of the invention may be intended for rectal administration, in the form, for example, of a suppository, which will melt in the rectum and release the drug.
  • the composition for rectal administration may contain an oleaginous base as a suitable nonirritating excipient.
  • bases include, without limitation, lanolin, cocoa butter and polyethylene glycol.
  • the combination-therapy pharmaceutical composition of the invention may include various materials, which modify the physical form of a solid or liquid dosage unit.
  • the composition may include materials that form a coating shell around the active ingredients.
  • the materials that form the coating shell are typically inert, and may be selected from, for example, sugar, shellac, and other enteric coating agents.
  • the active ingredients may be encased in a gelatin capsule.
  • the combination-therapy pharmaceutical composition of the invention in solid or liquid form may include an agent that binds to the compound of the invention and thereby assists in the delivery of the compound.
  • Suitable agents that may act in this capacity include a monoclonal or polyclonal antibody, a protein or a liposome.
  • the combination-therapy pharmaceutical composition of the invention may consist of dosage units that can be administered as an aerosol.
  • aerosol is used to denote a variety of systems ranging from those of colloidal nature to systems consisting of pressurized packages. Delivery may be by a liquefied or compressed gas or by a suitable pump system that dispenses the active ingredients. Aerosols of compounds of the invention may be delivered in single phase, bi-phasic, or th-phasic systems in order to deliver the active ingredient(s). Delivery of the aerosol includes the necessary container, activators, valves, subcontainers, and the like, which together may form a kit. One skilled in the art, without undue experimentation may determine preferred aerosols.
  • the combination-therapy pharmaceutical compositions of the invention may be prepared by methodology well known in the pharmaceutical art.
  • a combination-therapy pharmaceutical composition intended to be administered by injection can be prepared by combining a compound of the invention with sterile, distilled water so as to form a solution.
  • a surfactant may be added to facilitate the formation of a homogeneous solution or suspension.
  • Surfactants are compounds that non-covalently interact with the compound of the invention so as to facilitate dissolution or homogeneous suspension of the compound in the aqueous delivery system.
  • the combination-therapy pharmaceutical compositions of the invention can be formulated so as to provide quick, sustained or delayed release of the active ingredient after administration to the patient by employing procedures known in the art.
  • Controlled release drug delivery systems include osmotic pump systems and dissolutional systems containing polymer-coated reservoirs or drug-polymer matrix formulations. Examples of controlled release systems are given in U.S. Pat. Nos. 3,845,770 and 4,326,525 and in P. J. Kuzma et al, Regional Anesthesia 22 (6): 543- 551 (1997), all of which are incorporated herein by reference.
  • the combination-therapy pharmaceutical compositions of the invention can also be delivered through intra-nasal drug delivery systems for local, systemic, and nose-to-brain medical therapies.
  • Controlled Particle Dispersion (CPD)TM technology traditional nasal spray bottles, inhalers or nebulizers are known by those skilled in the art to provide effective local and systemic delivery of drugs by targeting the olfactory region and paranasal sinuses.
  • BIOLOGICAL ASSAYS Various techniques are known in the art for testing the effectiveness of the combination therapies of the invention. In order that the invention described herein may be more fully understood, the following biological assays are set forth. It should be understood that these examples are for illustrative purposes only and are not to be construed as limiting this invention in any manner.
  • This assay was conducted to determine if a single oral dose of 75 tng/Kg Lansoprazole or 50 mg/Kg of Compound A (i.e. a DMT1 inhibitor) can block the observed increase of serum iron in a dose dependent manner in a previously developed model of iron deficiency in Sprague Dawley rats when a 1 mg/Kg oral iron challenge is given 2 hours following drug dosing.
  • Compound A i.e. a DMT1 inhibitor
  • This assay was also conducted to determine if a combination therapy consisting of an oral dose of 15 mg/Kg Lansoprazole and 10 mg/Kg Compound A or an oral dose of 40 mg/Kg Lansoprazole and 25 mg/Kg Compound A can block the observed increase of serum iron in a dose dependent manner in a previously developed model of iron deficiency in Sprague Dawley rats when a 1 mg/Kg oral iron challenge is given 2 hours following dosing of the active ingredients.
  • FeSO 4 « 7H 2 O crystals were flushed with nitrogen following use to prevent oxidation.
  • 0.25 mg/mL FeSO 4 *7H 2 O was formulated as 0.325 g in 50 mL H 2 O.
  • Dose volume 1 mL/Kg (1 mg/Kg of FeSO 4 )
  • the iron-deficient rats responded to the oral iron challenge by absorbing iron rapidly, such that the vehicle for Lansoprazole and the vehicle for Compound A both resulted in an 11 -fold increase in serum iron within 1 hour of the iron challenge.
  • This assay was also conducted to determine if a combination therapy consisting of an oral dose of 15 mg/Kg Lansoprazole and 10 mg/Kg Compound A, an oral dose of 40 mg/Kg Lansoprazole and 25 mg/Kg Compound A, or an oral dose of 75 mg/Kg Lansoprazole and 50 mg/Kg Compound A can block the observed increase of serum iron in a dose dependent manner in a previously developed model of iron deficiency in Sprague Dawley rats when a 1 mg/Kg oral iron challenge is given 2 hours following dosing of the active ingredients.
  • FeSO 4 *7H 2 O crystals were flushed with nitrogen following use to prevent oxidation. 0.25 mg/mL FeSO 4 *7H 2 O was formulated as 0.325 g in 50 ml_ H 2 O. Dose volume: 1 mL/Kg (1 mg/Kg of FeSO 4 )
  • the iron-deficient rats responded to the oral iron challenge by absorbing iron rapidly, such that the vehicle for Lansoprazole and the vehicle for Compound A both resulted in an 4-fold increase in serum iron within 1 hour of the iron challenge.
  • oral administration of 15 mg/Kg Lansoprazole alone blocked iron absorption by approximately 4% when compared to the oral administration of the vehicle alone.
  • Oral administration of 40 mg/Kg Lansoprazole alone blocked iron absorption by approximately 27% when compared to the oral administration of the vehicle alone.
  • BIOLOGICAL ASSAY 3 The Effect of a Combination Therapy on the Acute Iron Deficiency Rat Model
  • This assay was conducted to determine if a single oral dose of 75 mg/Kg Omeprazole or 50 mg/Kg of Compound A (i.e. a DMT1 inhibitor) can block the observed increase of serum iron in a dose dependent manner in a previously developed model of iron deficiency in Sprague Dawley rats when a 1 mg/Kg oral iron challenge is given 2 hours following drug dosing.
  • Compound A i.e. a DMT1 inhibitor
  • This assay was also conducted to determine if a combination therapy consisting of an oral dose of 40 mg/Kg Omeprazole and 25 mg/Kg Compound A can block the observed increase of serum iron in a dose dependent manner in a previously developed model of iron deficiency in Sprague Dawley rats when a 1 mg/Kg oral iron challenge is given 2 hours following dosing of the active ingredients.
  • Dose volume 4 mL/Kg. Dose: 40 or 75 mg/Kg.
  • FeSO 4 *7H 2 O crystals were flushed with nitrogen following use to prevent oxidation. 0.25 mg/mL FeSO 4 *7H 2 O was formulated as 0.325 g in 50 ml_ H 2 O. Dose volume: 1 mL/Kg (1 mg/Kg of FeSO 4 )
  • oral administration of a combination of 40 mg/Kg Omeprazole and 25 mg/Kg Compound A blocked iron absorption by approximately 69% when compared to the oral administration of the Omeprazole vehicle.
  • Oral administration of only 75 mg/Kg Omeprazole blocked iron absorption by approximately 52% when compared to the oral administration of only the vehicle.
  • Oral administration of only 50 mg/Kg Compound A blocked iron absorption by approximately 55% when compared to the oral administration of only the vehicle.

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Abstract

This invention is directed to combination therapies to treat iron overload disorders utilizing inhibitors of divalent metal transporter-1 (DMT1 ), proton pump inhibitors (PPIs), calcium channel blockers and iron chelators.

Description

COMBINATION THERAPY FOR TREATING IRON DISORDERS
FIELD OF THE INVENTION
The present invention is directed to combination therapies for the treatment of iron overload disorders utilizing inhibitors of divalent metal transporter-1 (DMT1 ), proton pump inhibitors (PPIs), iron chelators and calcium channel blockers. Methods of using such combination therapies in the treatment of iron overload disorders and other disorders and methods of manufacture of such combination therapy are also provided.
BACKGROUND OF THE INVENTION Defects in iron metabolism contributing to iron overload represent a chronic health problem in adults. Iron overload is observed in diseases such as hemochromatosis, thalassemia and certain forms of anemia. In addition, iron overload can be caused by repeated blood transfusions outside of the treatment of these disorders. Iron overload can result in cirrhosis, hepatoma, arthritis, hypogonadism, diabetes and cardiomyopathy. Current therapies for iron overload include phlebotomy and treatment with chelators. However, given safety, side effects and compliance issues with both of these treatments, a market exists for safe drugs or combination therapies that act to block iron absorption in the gut and increase iron excretion.
The use of pharmaceutically acceptable iron chelators, such as Deferasirox, Deferoxamine and Deferiprone, in treating transfusional iron overload is well known. The efficacy of these iron chelators can be limited by dose dependent severe side effects particularly when administered at the higher dosages. These side effects can include, for example, renal impairment, liver toxicity and cytopenias. If these side effects are observed it is recommended that the chelation therapy be discontinued or the daily dosage of the chelator be reduced. This results in less efficacy as measured by the amount of iron removed from the body. Recently, due to the side effects and dosing constraints associated with each of these chelators, combination therapy, wherein two or more chelators are administered, has been proposed. See, for example, Kontoghiorghes, GJ. , Hemoglobin (2008), Vol. 32., Nos. 1-2, pp. 1-15. A recent study described the administration of certain proton pump inhibitors, i.e., omeprazole and lansoprazole, to patients with hereditary hemochromatosis. The administration of the proton pump inhibitors inhibited the absorption of non-heme iron in the gut (see, Hutchinson, C. et a/., Gut (2007), Vol. 56, No. 9, pp. 1291-1295). The postulated mechanism was that a higher pH in the gut acted to decrease the solubilization of non-heme iron and subsequently to decrease its absorption. Similarly, omeprazole resulted in a 50% decrease in ferrous iron absorption in rats on an iron- deficient diet (see, Golubov, J. et a/., Dig. Dis. Sci. (1991 ), Vol. 36, No. 4, pp. 405-408). The dose of omeprazole employed produced an effect on gastric acid output in rats similar to that produced by a dose recommended for the treatment of human gastrointestinal diseases.
Another recent study described the administration of proton pump inhibitors and Plavix, a commonly prescribed medication to prevent unwanted blood clots that occur after a recent heart attack or stroke. The study showed that commonly used proton pump inhibitors (e.g. lansoprazole, esomeprazole, omeprazole and pantoprazole) increased the risk of heart attacks and strokes by 50 percent among cardiac patients taking Plavix (see, Norgard , N. et a/., Ann. Pharmacother. (2009), published on-line on 2009 May 26). Therefore, it would appear that lengthy exposure to clinical dosages of proton pump inhibitors has serious liabilities. Other long term side effects of proton pump inhibitors include increased risks of colon cancer, gastric cancer, infection with C. difficile, respiratory infections and bone fracture.
It has been recently demonstrated that certain calcium channel blockers, such as nifedipine (dimethyl 1 ,4-dihydro-2,6-dimethyl-4-(o-nitrophenyl)-3,5- pyridinedicarboxylate), are a new pharmacological therapy useful for the treatment of iron overload disorders. For example, the administration of nifedipine increases DMT1 -mediated cellular iron transport and enhances urinary iron excretion (see, Ludwiczek, et al., (2007), Vol. 13, No. 4, pp. 448-454). The mechanism by which nifedipine causes this effect is by prolonging the iron transporting activity of DMT1 in the duodenum and thus likely increase urinary iron output by prolonging the iron transporting activity of DMT1 in the kidney.
DMT1 is expressed in the duodenum and regulates absorption of iron. Recently, a series of compounds has been developed as DMT1 inhibitors (see, for example, PCT Published Application No. WO 2008/109840; PCT Published Application No. WO 2008/115999; PCT Published Application No. WO 2008/118790; PCT
Published Application No. WO 2008/121861 ; and PCT Published Application No. WO 2008/151288; the entire disclosures of which are incorporated in full by reference herein).
The use of iron chelators in combination with a proton pump inhibitor (PPI), a calcium channel blocker or a DMT1 inhibitor, or any combination thereof, is not known. A PPI, a DMT1 inhibitor or a calcium channel blocker could work additively with chelation therapy to remove the excess iron that is in the body by blocking the uptake of iron from the intestine into the body or by increasing the urinary iron output from the kidney. This combination may increase efficacy and improve iron serum and tissue control in iron-overloaded patients or could lead to reduction in the dosage or frequency of dosing chelators which could lead to safer treatments for patients by reducing the chelation treatment-induced side effects.
Accordingly, there is an unmet medical need to treat iron overload disorders, preferably primary iron overload and transfusional iron overload in mammals, preferably in humans, using a combination of DMT1 inhibitors with iron chelators, calcium channel blockers or PPIs, or any combination thereof, or PPIs with iron chelators.
SUMMARY OF THE INVENTION
The present invention is directed to combination therapies for treating an iron overload disorder in a mammal, wherein the combination therapies comprise administering to the mammal a therapeutically effective amount of a DMT1 inhibitor and a therapeutically effective amount of a proton pump inhibitor, and, optionally, a therapeutically effective amount of a pharmaceutically acceptable iron chelator, and, optionally a therapeutically effective amount of a calcium channel blocker. Alternatively, the present invention is directed to combination therapies for treating an iron overload disorder in a mammal, wherein the combination therapies comprise administering to the mammal a therapeutically effective amount of a DMT1 inhibitor and a therapeutically effective amount of a pharmaceutically acceptable iron chelator and, optionally, a therapeutically effective amount of a calcium channel blocker.
Alternatively, the present invention is directed to combination therapies for treating an iron overload disorder in a mammal, wherein the combination therapies comprise administering to the mammal a therapeutically effective amount of a DMT1 inhibitor and a therapeutically effective amount of a calcium channel blocker, and, optionally, a therapeutically effective amount of a proton pump inhibitor.
Alternatively, the present invention is directed to combination therapies for treating an iron overload disorder in a mammal, wherein the combination therapies comprise administering to the mammal a therapeutically effective amount of a pharmaceutically acceptable iron chelator and a therapeutically effective amount of a calcium channel blocker and/or a therapeutically effective amount of a proton pump inhibitor.
Alternatively, the present invention is directed to combination therapies for treating an iron overload disorder in a mammal, wherein the combination therapies comprise administering to the mammal a therapeutically effective amount of a proton pump inhibitor and a therapeutically effective amount of a calcium channel blocker.
These combination therapies may be administered prior to, concurrently with, or subsequent to a phlebotomy or other therapeutic erythrocytapheresis treatment or at all of these timepoints. Accordingly, in one aspect, this invention is directed to a method of treating an iron overload disorder in a mammal, wherein the method comprises administering to the mammal in need thereof a therapeutically effective amount of a DMT1 inhibitor and a therapeutically effective amount of a pharmaceutically acceptable iron chelator. In another aspect, this invention is directed to a method of treating an iron overload disorder in a mammal, wherein the method comprises administering to the mammal in need thereof a therapeutically effective amount of a DMT1 inhibitor and a therapeutically effective amount of a proton pump inhibitor.
In another aspect, this invention is directed to a method of treating an iron overload disorder in a mammal, wherein the method comprises administering to the mammal in need thereof a therapeutically effective amount of a DMT1 inhibitor and a therapeutically effective amount of a calcium channel blocker.
In another aspect, this invention is directed to a method of treating an iron overload disorder in a mammal, wherein the method comprises administering to the mammal in need thereof a therapeutically effective amount of a DMT1 inhibitor, a therapeutically effective amount of a pharmaceutically acceptable iron chelator, and a therapeutically effective amount of a proton pump inhibitor.
In another aspect, this invention is directed to a method of treating an iron overload disorder in a mammal, wherein the method comprises administering to the mammal in need thereof a therapeutically effective amount of a DMT1 inhibitor, a therapeutically effective amount of a pharmaceutically acceptable iron chelator, and a therapeutically effective amount of a calcium channel blocker.
In another aspect, this invention is directed to a method of treating an iron overload disorder in a mammal, wherein the method comprises administering to the mammal in need thereof a therapeutically effective amount of a DMT1 inhibitor, a therapeutically effective amount of a proton pump inhibitor, and a therapeutically effective amount of a calcium channel blocker.
In another aspect, this invention is directed to a method of treating an iron overload disorder in a mammal, wherein the method comprises administering to the mammal in need thereof a therapeutically effective amount of a DMT1 inhibitor, a therapeutically effective amount of a calcium channel blocker, a therapeutically effective amount of a proton pump inhibitor, and a therapeutically effective amount of a pharmaceutically acceptable iron chelator.
In another aspect, this invention is directed to a method for treating an iron overload disorder in a mammal, wherein the combination therapy comprises the administration to the mammal in need thereof of a therapeutically effective amount of a proton pump inhibitor and a therapeutically effective amount of a pharmaceutically acceptable iron chelator.
In another aspect, this invention is directed to a method for treating an iron overload disorder in a mammal, wherein the combination therapy comprises the administration to the mammal in need thereof of a therapeutically effective amount of a proton pump inhibitor, a therapeutically effective amount of a pharmaceutically acceptable iron chelator and a therapeutically effective amount of a calcium channel blocker.
In another aspect, this invention is directed to a method for treating an iron overload disorder in a mammal, wherein the combination therapy comprises the administration to the mammal in need thereof of a therapeutically effective amount of a proton pump inhibitor and a therapeutically effective amount of a calcium channel blocker.
The various aspects of the invention are described in more detail below. DETAILED DESCRIPTION OF THE INVENTION
DEFINITIONS
Certain chemical groups named herein may be preceded by a shorthand notation indicating the total number of carbon atoms that are to be found in the indicated chemical group. For example; C7-C12alkyl describes an alkyl group, as defined below, having a total of 7 to 12 carbon atoms, and C4-C12cycloalkylalkyl describes a cycloalkylalkyl group, as defined below, having a total of 4 to 12 carbon atoms. The total number of carbons in the shorthand notation does not include carbons that may exist in substituents of the group described. In addition to the foregoing, as used in the specification and appended claims, unless specified to the contrary, the following terms have the meaning indicated:
"Amino" refers to the -NH2 radical.
"Cyano" refers to the -CN radical. "Hydroxy" refers to the -OH radical.
"Imino" refers to the =NH substituent.
"Nitro" refers to the -NO2 radical.
"Oxo" refers to the =0 substituent.
"Thioxo" refers to the =S substituent. "Thfluoromethyl" refers to the -CF3 radical.
"Alkyl" refers to a straight or branched hydrocarbon chain radical consisting solely of carbon and hydrogen atoms, containing no unsaturation, having from one to twelve carbon atoms, preferably one to eight carbon atoms or one to six carbon atoms, and which is attached to the rest of the molecule by a single bond, e.g., methyl, ethyl, n-propyl, 1-methylethyl (/so-propyl), n-butyl, n-pentyl, 1 ,1-dimethylethyl (f-butyl),
3-methylhexyl, 2-methylhexyl, and the like. Unless stated otherwise specifically in the specification, an alkyl group may be optionally substituted by one of the following groups: alkyl, alkenyl, halo, haloalkenyl, cyano, nitro, aryl, cycloalkyl, heterocyclyl, heteroaryl, oxo, trimethylsilanyl, -OR30, -OC(O)-R30, -N(R30)2> -C(O)R30, -C(O)OR30, -C(O)N(R30)2, -N(R30)C(O)OR32, -N(R30)C(O)R32, -N(R30)S(O),R32 (where t is 1 to 2), -S(O)1OR32 (where t is 1 to 2), -S(O)PR32 (where p is O to 2), and -S(O),N(R30)2 (where t is 1 to 2) where each R30 is independently hydrogen, alkyl, haloalkyl, cycloalkyl, cycloalkylalkyl, aryl, aralkyl, heterocyclyl, heterocyclylalkyl, heteroaryl or heteroarylalkyl; and each R32 is alkyl, haloalkyl, cycloalkyl, cycloalkylalkyl, aryl, aralkyl, heterocyclyl, heterocyclylalkyl, heteroaryl or heteroarylalkyl.
"Alkenyl" refers to a straight or branched hydrocarbon chain radical group consisting solely of carbon and hydrogen atoms, containing at least one double bond, having from two to twelve carbon atoms, preferably two to eight carbon atoms and which is attached to the rest of the molecule by a single bond, e.g., ethenyl, prop-1-enyl, but-1-enyl, pent-1-enyl, penta-1 ,4-dienyl, and the like. Unless stated otherwise specifically in the specification, an alkenyl group may be optionally substituted by one of the following groups: alkyl, alkenyl, halo, haloalkenyl, cyano, nitro, aryl, cycloalkyl, heterocyclyl, heteroaryl, oxo, trimethylsilanyl, -OR30, -OC(O)-R30, -N(R30)2, -C(O)R30, -C(O)OR30, -C(O)N(R30)2, -N(R30)C(O)OR32, -N(R30)C(O)R32, -N(R30)S(O),R32 (where t is 1 to 2), -S(O)tOR32 (where t is 1 to 2), -S(O)PR32 (where p is 0 to 2), and -S(O)tN(R30)2 (where t is 1 to 2) where each R30 is independently hydrogen, alkyl, haloalkyl, cycloalkyl, cycloalkylalkyl, aryl, aralkyl, heterocyclyl, heterocyclylalkyl, heteroaryl or heteroarylalkyl; and each R32 is alkyl, haloalkyl, cycloalkyl, cycloalkylalkyl, aryl, aralkyl, heterocyclyl, heterocyclylalkyl, heteroaryl or heteroarylalkyl. "Alkynyl" refers to a straight or branched hydrocarbon chain radical group comprising solely of carbon and hydrogen atoms, containing at least one triple bond, optionally containing at least one double bond, having from two to twelve carbon atoms, preferably two to eight carbon atoms and which is attached to the rest of the molecule by a single bond, for example, ethynyl, propynyl, butynyl, pentynyl, hexynyl, and the like. Unless stated otherwise specifically in the specification, an alkynyl group may be optionally substituted by one or more of the following substituents: alkyl, alkenyl, halo, haloalkenyl, cyano, nitro, aryl, cycloalkyl, heterocyclyl, heteroaryl, oxo, trimethylsilanyl, -OR30, -OC(O)-R30, -N(R30)2, -C(O)R30, -C(O)OR30, -C(O)N(R30)2, -N(R30)C(O)OR32, -N(R30)C(O)R32, -N(R30)S(O)(R32 (where t is 1 to 2), -S(O)1OR32 (where t is 1 to 2), -S(O)PR32 (where p is O to 2), and -S(O)tN(R30)2 (where t is 1 to 2) where each R30 is independently hydrogen, alkyl, haloalkyl, cycloalkyl, cycloalkylalkyl, aryl, aralkyl, heterocyclyl, heterocyclylalkyl, heteroaryl or heteroarylalkyl; and each R32 is alkyl, haloalkyl, cycloalkyl, cycloalkylalkyl, aryl, aralkyl, heterocyclyl, heterocyclylalkyl, heteroaryl or heteroarylalkyl. "Alkylene" or "alkylene chain" refers to a straight or branched divalent hydrocarbon chain linking the rest of the molecule to a radical group, consisting solely of carbon and hydrogen, containing no unsaturation and having from one to twelve carbon atoms, e.g., methylene, ethylene, propylene, n-butylene, and the like. The alkylene chain is attached to the rest of the molecule through a single bond and to the radical group through a single bond. The points of attachment of the alkylene chain to the rest of the molecule and to the radical group can be through one carbon or any two carbons within the chain. Unless stated otherwise specifically in the specification, an alkylene chain may be optionally substituted by one of the following groups: alkyl, alkenyl, halo, haloalkenyl, cyano, nitro, aryl, cycloalkyl, heterocyclyl, heteroaryl, oxo, trimethylsilanyl, -OR30, -OC(O)-R30, -N(R30)2, -C(O)R30, -C(O)OR30, -C(O)N(R30)2, -N(R30)C(O)OR32, -N(R30)C(O)R32, -N(R30)S(O)tR32 (where t is 1 to 2), -S(O)1OR32 (where t is 1 to 2), -S(O)PR32 (where p is O to 2), and -S(O),N(R30)2 (where t is 1 to 2) where each R30 is independently hydrogen, alkyl, haloalkyl, cycloalkyl, cycloalkylalkyl, aryl, aralkyl, heterocyclyl, heterocyclylalkyl, heteroaryl or heteroarylalkyl; and each R32 is alkyl, haloalkyl, cycloalkyl, cycloalkylalkyl, aryl, aralkyl, heterocyclyl, heterocyclylalkyl, heteroaryl or heteroarylalkyl.
"Alkenylene" or "alkenylene chain" refers to a straight or branched divalent hydrocarbon chain linking the rest of the molecule to a radical group, consisting solely of carbon and hydrogen, containing at least one double bond and having from two to twelve carbon atoms, e.g., ethenylene, propenylene, n-butenylene, and the like. The alkenylene chain is attached to the rest of the molecule through a single bond and to the radical group through a double bond or a single bond. The points of attachment of the alkenylene chain to the rest of the molecule and to the radical group can be through one carbon or any two carbons within the chain. Unless stated otherwise specifically in the specification, an alkenylene chain may be optionally substituted by one of the following groups: alkyl, alkenyl, halo, haloalkenyl, cyano, nitro, aryl, cycloalkyl, heterocyclyl, heteroaryl, oxo, trimethylsilanyl, -OR30, -OC(O)-R30, -N(R30)2, -C(O)R30, -C(O)OR30, -C(O)N(R30)2, -N(R30)C(O)OR32, -N(R30)C(O)R32, -N(R30)S(O)tR32 (where t is 1 to 2), -S(O)1OR32 (where t is 1 to 2), -S(O)PR32 (where p is O to 2), and -S(O)tN(R30)2 (where t is 1 to 2) where each R30 is independently hydrogen, alkyl, haloalkyl, cycloalkyl, cycloalkylalkyl, aryl, aralkyl, heterocyclyl, heterocyclylalkyl, heteroaryl or heteroarylalkyl; and each R32 is alkyl, haloalkyl, cycloalkyl, cycloalkylalkyl, aryl, aralkyl, heterocyclyl, heterocyclylalkyl, heteroaryl or heteroarylalkyl.
"Alkoxy" refers to a radical of the formula -OR3 where Ra is an alkyl radical as defined above containing one to twelve carbon atoms. The alkyl part of the alkoxy radical may be optionally substituted as defined above for an alkyl radical.
"Alkoxyalkyl" refers to a radical of the formula -Rb-O-Ra where Rb is an alkylene chain as defined above and R3 is an alkyl radical as defined above. The oxygen atom may be bonded to any carbon in the alkylene chain and in the alkyl radical. The alkyl part of the alkoxyalkyl radical may be optionally substituted as defined above for an alkyl group. The alkylene chain part of the alkoxyalkyl radical may be optionally substituted as defined above for an alkylene chain.
"Aryl" refers to a hydrocarbon ring system radical comprising hydrogen, 6 to 18 carbon atoms and at least one aromatic ring. For purposes of this invention, the aryl radical may be a monocyclic, bicyclic, tricyclic or tetracyclic ring system, which may included fused or bridged ring systems. Aryl radicals include, but are not limited to, aryl radicals derived from aceanthrylene, acenaphthylene, acephenanthrylene, anthracene, azulene, benzene, chrysene, fluoranthene, fluorene, as-indacene, s-indacene, indane, indene, naphthalene, phenalene, phenanthrene, pleiadene, pyrene, and triphenylene. Unless stated otherwise specifically in the specification, the term "aryl" or the prefix "ar-" (such as in "aralkyl") is meant to include aryl radicals optionally substituted by one or more substituents independently selected from the group consisting of alkyl, alkenyl, halo, haloalkyl, haloalkenyl, cyano, nitro, aryl, aralkyi, heteroaryl, heteroarylalkyl, -R31-OR30, -R31-OC(O)-R30, -R31-N(R30)2, -R31 -C(O)R30, -R31 -C(O)OR30, -R31-C(O)N(R30)2, -R31-N(R30)C(O)OR32, -R31-N(R30)C(O)R32,
-R31-N(R30)S(O)tR32 (where t is 1 to 2), -R31-N=C(OR30)R30, -R31 -S(O)1OR32 (where t is 1 to 2), -R31-S(O)PR32 (where p is 0 to 2), and -R31-S(O)tN(R30)2 (where t is 1 to 2) where each R30 is independently hydrogen, alkyl, haloalkyl, cycloalkyl, cycloalkylalkyl, aryl, aralkyl, heterocyclyl, heterocyclylalkyl, heteroaryl or heteroarylalkyl; each R31 is independently a direct bond or a straight or branched alkylene or alkenylene chain; and each R32 is alkyl, haloalkyl, cycloalkyl, cycloalkylalkyl, aryl, aralkyl, heterocyclyl, heterocyclylalkyl, heteroaryl or heteroarylalkyl.
"Aralkyl" refers to a radical of the formula -Rb-Rc where Rb is an alkylene chain as defined above and Rc is one or more aryl radicals as defined above, for example, benzyl, diphenylmethyl and the like. The alkylene chain part of the aralkyl radical may be optionally substituted as described above for an alkylene chain. The aryl part of the aralkyl radical may be optionally substituted as described above for an aryl group.
"Aralkenyl" refers to a radical of the formula -Rd-Rc where Rd is an alkenylene chain as defined above and Rc is one or more aryl radicals as defined above. The aryl part of the aralkenyl radical may be optionally substituted as described above for an aryl group. The alkenylene chain part of the aralkenyl radical may be optionally substituted as defined above for an alkenylene group.
"Cycloalkyl" refers to a stable non-aromatic monocyclic or polycyclic hydrocarbon radical consisting solely of carbon and hydrogen atoms, which may include fused or bridged ring systems, having from three to fifteen carbon atoms, preferably having from three to ten carbon atoms, and which is saturated or unsaturated and attached to the rest of the molecule by a single bond. Monocyclic radicals include, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptly, and cyclooctyl. Polycyclic radicals include, for example, adamantyl, norbornyl, decalinyl, 7,7-dimethyl-bicyclo[2.2.1]heptanyl, and the like. Unless otherwise stated specifically in the specification, the term "cycloalkyl" is meant to include cycloalkyl radicals which are optionally substituted by one or more substituents independently selected from the group consisting of alkyl, alkenyl, halo, haloalkyl, haloalkenyl, cyano, nitro, oxo, aryl, aralkyl, cycloalkyl, cycloalkylalkyl, heterocyclyl, heterocyclylalkyl, heteroaryl, heteroarylalkyl, -R31 -OR30, -R31-OC(O)-R30, -R31-N(R30)2, -R31-C(O)R30, -R31-C(O)OR30, -R31-C(O)N(R30)2, -R31-N(R30)C(O)OR32, -R31-N(R30)C(O)R32, -R31-N(R30)S(O)tR32 (where t is 1 to 2), -R31-N=C(OR30)R30, -R31 -S(O)tOR32 (where t is 1 to 2), -R31-S(O)PR32 (where p is 0 to 2), and -R31-S(O)tN(R30)2 (where t is 1 to 2) where each R30 is independently hydrogen, alkyl, haloalkyl, cycloalkyl, cycloalkylalkyl, aryl, aralkyl, heterocyclyl, heterocyclylalkyl, heteroaryl or heteroarylalkyl; each R31 is independently a direct bond or a straight or branched alkylene or alkenylene chain; and each R32 is alkyl, haloalkyl, cycloalkyl, cycloalkylalkyl, aryl, aralkyl, heterocyclyl, heterocyclylalkyl, heteroaryl or heteroarylalkyl. "Cycloalkylalkyl" refers to a radical of the formula -RbRg where Rb is an alkylene chain as defined above and R9 is a cycloalkyl radical as defined above. The alkylene chain and the cycloalkyl radical may be optionally substituted as defined above. "Fused" refers to any ring structure described herein which is fused to an existing ring structure in the compounds of the invention. When the fused ring is a heterocyclyl ring or a heteroaryl ring, any carbon atom on the existing ring structure which becomes part of the fused heterocyclyl ring or the fused heteroaryl ring may be replaced with a nitrogen atom.
"Halo" refers to bromo, chloro, fluoro or iodo.
"Haloalkyl" refers to an alkyl radical, as defined above, that is substituted by one or more halo radicals, as defined above, e.g., trifluoromethyl, difluoromethyl, trichloromethyl, 2,2,2-trifluoroethyl, 1-fluoromethyl-2-fluoroethyl, 3-bromo-2-fluoropropyl, i-bromomethyl-2-bromoethyl, and the like. The alkyl part of the haloalkyl radical may be optionally substituted as defined above for an alkyl group. "Haloalkoxy" refers to a radical of the formula -ORj where Rj is a haloalkyl radical as defined above, e.g., trifluoromethoxy, difluoromethoxy, trichloromethoxy, 2,2,2-trifluoroethoxy, 1 -fluoromethyl-2-fluoroethoxyl, 3-bromo-2-fluoropropoxy, i-bromomethyl-2-bromoethoxy, and the like. The alkyl part of the haloalkoxy radical may be optionally substituted as defined above for an alkyl group.
"Haloalkenyl" refers to an alkenyl radical, as defined above, that is substituted by one or more halo radicals, as defined above. The alkenyl part of the haloalkyl radical may be optionally substituted as defined above for an alkenyl group.
"Heterocyclyl" refers to a stable 3- to 18-membered non-aromatic ring radical which consists of two to twelve carbon atoms and from one to six heteroatoms selected from the group consisting of nitrogen, oxygen and sulfur. Unless stated otherwise specifically in the specification, the heterocyclyl radical may be a monocyclic, bicyclic, tricyclic or tetracyclic ring system, which may include fused or bridged ring systems; and the nitrogen, carbon or sulfur atoms in the heterocyclyl radical may be optionally oxidized; the nitrogen atom may be optionally quatemized; and the heterocyclyl radical may be partially or fully saturated. Examples of such heterocyclyl radicals include, but are not limited to, dioxolanyl, thienyl[1 ,3]dithianyl, decahydroisoquinolyl, imidazolinyl, imidazolidinyl, isothiazolidinyl, isoxazolidinyl, morpholinyl, octahydroindolyl, octahydroisoindolyl, 2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxopyrrolidinyl, oxazolidinyl, piperidinyl, piperazinyl, 4-piperidonyl, pyrrolidinyl, pyrazolidinyi, quinuclidinyl, thiazolidinyl, tetrahydrofuryl, trithianyl, tetrahydropyranyl, thiomorpholinyl, thiamorpholinyl, 1-oxo-thiomorpholinyl, and 1 ,1-dioxo-thiomorpholinyl. Unless stated otherwise specifically in the specification, the term "heterocyclyl" is meant to include heterocyclyl radicals as defined above which are optionally substituted by one or more substituents selected from the group consisting of alkyl, alkenyl, halo, haloalkyl, haloalkenyl, cyano, oxo, thioxo, nitro, aryl, aralkyl, cycloalkyl, cycloalkylalkyl, heterocyclyl, heterocyclylalkyl, heteroaryl, heteroarylalkyl, -R31-OR30, -R31-OC(O)-R30, -R31-N(R30)2, -R31-C(O)R30, -R31 -C(O)OR30, -R31-C(O)N(R30)2, -R31-N(R30)C(O)OR32, -R31-N(R30)C(O)R32, -R31-N(R30)S(O)tR32 (where t is 1 to 2), -R31-N=C(OR30)R30, -R31 -S(O)1OR32 (where t is 1 to 2), -R31-S(O)PR32 (where p is 0 to 2), and -R31-S(O)tN(R30)2 (where t is 1 to 2) where each R30 is independently hydrogen, alkyl, alkenyl, haloalkyl, cycloalkyl, cycloalkylalkyl, aryl, aralkyl, heterocyclyl, heterocyclylalkyl, heteroaryl or heteroarylalkyl; each R31 is independently a direct bond or a straight or branched alkylene or alkenylene chain; and each R32 is alkyl, alkenyl, haloalkyl, cycloalkyl, cycloalkylalkyl, aryl, aralkyl, heterocyclyl, heterocyclylalkyl, heteroaryl or heteroarylalkyl. "Λ/-heterocyclyl" refers to a heterocyclyl radical as defined above containing at least one nitrogen and where the point of attachment of the heterocyclyl radical to the rest of the molecule is through a nitrogen atom in the heterocyclyl radical. An Λ/-heterocyclyl radical may be optionally substituted as described above for heterocyclyl radicals. "Heterocyclylalkyl" refers to a radical of the formula -RbRh where Rb is an alkylene chain as defined above and Rh is a heterocyclyl radical as defined above, and if the heterocyclyl is a nitrogen-containing heterocyclyl, the heterocyclyl may be attached to the alkyl radical at the nitrogen atom. The alkylene chain of the heterocyclylalkyl radical may be optionally substituted as defined above for an alkylene chain. The heterocyclyl part of the heterocyclylalkyl radical may be optionally substituted as defined above for a heterocyclyl group.
"Heteroaryl" refers to a 5- to 14-membered ring system radical comprising hydrogen atoms, one to thirteen carbon atoms, one to six heteroatoms selected from the group consisting of nitrogen, oxygen and sulfur, and at least one aromatic ring. For purposes of this invention, the heteroaryl radical may be a monocyclic, bicyclic, tricyclic or tetracyclic ring system, which may include fused or bridged ring systems; and the nitrogen, carbon or sulfur atoms in the heteroaryl radical may be optionally oxidized; the nitrogen atom may be optionally quaternized. Examples include, but are not limited to, azepinyl, acridinyl, benzimidazolyl, benzthiazolyl, benzindolyl, benzodioxolyl, benzofuranyl, benzooxazolyl, benzothiazolyl, benzothiadiazolyl, benzo[£>][1 ,4]dioxepinyl, 1 ,4-benzodioxanyl, benzonaphthofuranyl, benzoxazolyl, benzodioxolyl, benzodioxinyl, benzopyranyl, benzopyranonyl, benzofuranyl, benzofuranonyl, benzothienyl (benzothiophenyl), benzotriazolyl, benzo[4,6]imidazo[1 ,2-a]pyridinyl, carbazolyl, cinnolinyl, dibenzofuranyl, dibenzothiophenyl, furanyl, furanonyl, isothiazolyl, imidazolyl, indazolyl, indolyl, indazolyl, isoindolyl, indolinyl, isoindolinyl, isoquinolyl, indolizinyl, isoxazolyl, naphthyridinyl, oxadiazolyl, 2-oxoazepinyl, oxazolyl, oxiranyl, 1-oxidopyridinyl, 1-oxidopyrimidinyl, 1-oxidopyrazinyl, 1-oxidopyridazinyl, 1-phenyl-1/-/-pyrrolyl, phenazinyl, phenothiazinyl, phenoxazinyl, phthalazinyl, pteridinyl, purinyl, pyrrolyl, pyrazolyl, pyridinyl, pyrazinyl, pyrimidinyl, pyridazinyl, pyrrolyl, quinazolinyl, quinoxalinyl, quinolinyl, quinuclidinyl, isoquinolinyl, tetrahydroquinolinyl, thiazolyl, thiadiazolyl, triazolyl, tetrazolyl, triazinyl, and thiophenyl (i.e. thienyl). Unless stated otherwise specifically in the specification, the term "heteroaryl" is meant to include heteroaryl radicals as defined above which are optionally substituted by one or more substituents selected from the group consisting of alky!, alkenyl, alkoxy, halo, haloalkyl, haloalkenyl, cyano, oxo, nitro, thioxo, aryl, aralkyl, cycloalkyl, cycloalkylalkyl, heterocyclyl, heterocyclylalkyl, heteroaryl, heteroarylalkyl, -R31-OR30, -R31 -OC(O)-R30, -R31-N(R30)2, -R31-C(O)R30, -R31-C(O)OR30, -R31-C(O)N(R30)2, -R31-N(R30)C(O)OR32, -R31-N(R30)C(O)R32, -R31-N(R30)S(O),R32 (where t is 1 to 2), -R31-N=C(OR30)R30, -R31 -S(O)1OR32 (where t is 1 to 2), -R31-S(O)PR32 (where p is 0 to 2), and
-R31-S(O)tN(R30)2 (where t is 1 to 2) where each R30 is independently hydrogen, alkyl, alkenyl, haloalkyl, cycloalkyl, cycloalkylalkyl, aryl, aralkyl, heterocyclyl, heterocyclylalkyl, heteroaryl or heteroarylalkyl; each R31 is independently a direct bond or a straight or branched alkylene or alkenylene chain; and each R32 is alkyl, alkenyl, haloalkyl, cycloalkyl, cycloalkylalkyl, aryl, aralkyl, heterocyclyl, heterocyclylalkyl, heteroaryl or heteroarylalkyl.
"Λ/-heteroaryl" refers to a heteroaryl radical as defined above containing at least one nitrogen and where the point of attachment of the heteroaryl radical to the rest of the molecule is through a nitrogen atom in the heteroaryl radical. An Λ/-heteroaryl radical may be optionally substituted as described above for heteroaryl radicals.
"Heteroarylalkyl" refers to a radical of the formula -RbRi where Rb is an alkylene chain as defined above and Ri is a heteroaryl radical as defined above. The heteroaryl part of the heteroarylalkyl radical may be optionally substituted as defined above for a heteroaryl group. The alkylene chain part of the heteroarylalkyl radical may be optionally substituted as defined above for an alkylene chain.
"Mammal" includes humans and both domestic animals such as laboratory animals and household pets, (e.g., cats, dogs, swine, cattle, sheep, goats, horses, rabbits), and non-domestic animals such as wildlife and the like.
"Optional" or "optionally" means that the subsequently described event of circumstances may or may not occur, and that the description includes instances where said event or circumstance occurs and instances in which it does not. For example, "optionally substituted aryl" means that the aryl radical may or may not be substituted and that the description includes both substituted aryl radicals and aryl radicals having no substitution. When a functional group is described as "optionally substituted," and in turn, substitutents on the functional group are also "optionally substituted" and so on, for the purposes of this invention, such iterations are limited to five, preferably such iterations are limited to two.
"Pharmaceutically acceptable iron chelator" includes any molecule capable of chelating non-heme iron in a mammal and which has been approved by an appropriate regulating authority, such as the United States Food and Drug Administration, as being acceptable for use in mammals, preferably in humans.
"Pharmaceutically acceptable carrier, diluent or excipient" includes without limitation any adjuvant, carrier, excipient, glidant, sweetening agent, diluent, preservative, dye/colorant, flavor enhancer, surfactant, wetting agent, dispersing agent, suspending agent, stabilizer, isotonic agent, solvent, or emulsifier which has been approved by an appropriate regulating authority, such as the United States Food and Drug Administration, as being acceptable for use in humans or domestic animals.
"Pharmaceutically acceptable salt" includes both acid and base addition salts.
"Pharmaceutically acceptable acid addition salt" refers to those salts which retain the biological effectiveness and properties of the free bases, which are not biologically or otherwise undesirable, and which are formed with inorganic acids such as, but are not limited to, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid and the like, and organic acids such as, but not limited to, acetic acid, 2,2-dichloroacetic acid, adipic acid, alginic acid, ascorbic acid, aspartic acid, benzenesulfonic acid, benzoic acid, 4-acetamidobenzoic acid, camphoric acid, camphor-10-sulfonic acid, capric acid, caproic acid, caprylic acid, carbonic acid, cinnamic acid, citric acid, cyclamic acid, dodecylsulfuric acid, ethane-1 ,2-disulfonic acid, ethanesulfonic acid, 2-hydroxyethanesulfonic acid, formic acid, fumaric acid, galactaric acid, gentisic acid, glucoheptonic acid, gluconic acid, glucuronic acid, glutamic acid, glutaric acid, 2-oxo-glutaric acid, glycerophosphoric acid, glycolic acid, hippuric acid, isobutyric acid, lactic acid, lactobionic acid, lauric acid, maleic acid, malic acid, malonic acid, mandelic acid, methanesulfonic acid, mucic acid, naphthalene-1 ,5- disulfonic acid, naphthalene-2-sulfonic acid, 1-hydroxy-2-naphthoic acid, nicotinic acid, oleic acid, orotic acid, oxalic acid, palmitic acid, pamoic acid, propionic acid, pyroglutamic acid, pyruvic acid, salicylic acid, 4-aminosalicylic acid, sebacic acid, stearic acid, succinic acid, tartaric acid, thiocyanic acid, p-toluenesulfonic acid, trifluoroacetic acid, undecylenic acid, and the like.
"Pharmaceutically acceptable base addition salt" refers to those salts which retain the biological effectiveness and properties of the free acids, which are not biologically or otherwise undesirable. These salts are prepared from addition of an inorganic base or an organic base to the free acid. Salts derived from inorganic bases include, but are not limited to, the sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, aluminum salts and the like. Preferred inorganic salts are the ammonium, sodium, potassium, calcium, and magnesium salts. Salts derived from organic bases include, but are not limited to, salts of primary, secondary, and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines and basic ion exchange resins, such as ammonia, isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, diethanolamine, ethanolamine, deanol, 2-dimethylaminoethanol, 2-diethylaminoethanol, dicyclohexylamine, lysine, arginine, histidine, caffeine, procaine, hydrabamine, choline, betaine, benethamine, benzathine, ethylenediamine, glucosamine, methylglucamine, theobromine, triethanolamine, tromethamine, purines, piperazine, piperidine, Λ/-ethylpiperidine, polyamine resins and the like. Particularly preferred organic bases are isopropylamine, diethylamine, ethanolamine, trimethylamine, dicyclohexylamine, choline and caffeine. A "pharmaceutical composition" refers to a formulation of a biologically active ingredient and a medium generally accepted in the art for the delivery of the biologically active ingredient to mammals, e.g., humans. Such a medium includes all pharmaceutically acceptable carriers, diluents or excipients therefor. "Therapeutically effective amount" refers to that amount of a biologically active ingredient which, when administered to a mammal, preferably a human, is sufficient to effect treatment, as defined below, of an iron overload disorder in the mammal, preferably a human. The amount of a biologically active ingredient which constitutes a "therapeutically effective amount" will vary depending on the active ingredient, the iron overload disorder and its severity, the manner of administration, and the age of the mammal to be treated, but can be determined routinely by one of ordinary skill in the art having regard to his own knowledge and to this disclosure. Preferably, for purposes of this invention, a "therapeutically effective amount" is that amount of an active ingredient sufficient to produce a negative iron balance in the mammal treated. "Treating" or "treatment", as used herein, covers the treatment of an iron overload disorder in a mammal, preferably a human and includes:
(i) preventing an iron overload disorder from occurring in the mammal;
(ii) inhibiting an iron overload disorder in a mammal, i.e., arresting its development; (iii) relieving an iron overload disorder in a mammal, i.e., causing a regression of the iron disorder or the disease or condition;
(iv) relieving the symptoms of an iron overload disorder in a mammal, i.e., relieving the symptoms without addressing the underlying iron overload disorder; or
(v) restoring and/or maintaining normal serum iron levels, transferrin saturation, serum ferritin, liver iron and/or bodily iron levels in a mammal having an iron overload disorder.
"Active ingredient" as used herein refers to a DMT1 inhibitor (either an agent directly inhibiting DMT1 or an agent that reduces the driving force for iron influx in enterocytes), a pharmaceutically acceptable iron chelator, a calcium channel blocker or a proton pump inhibitor utilized in the combination therapies of the invention.
As used herein, the terms "disease" and "condition" may be used interchangeably or may be different in that the particular malady or condition may not have a known causative agent (so that etiology has not yet been worked out) and it is therefore not yet recognized as a disease but only as an undesirable condition or syndrome, wherein a more or less specific set of symptoms have been identified by clinicians.
As used herein "combination therapy" refers to the administration of more than one active ingredient for the treatment of an iron overload disorder. Unless the context makes it clear otherwise, "combination therapy" may include the administration of any mixture or permutation of more than one active ingredient . Unless the context makes it clear otherwise, "combination therapy" may include simultaneous or sequential administration of two or more different active ingredients, in any order, one after the other. Unless the context makes clear otherwise, "combination therapy " may include dosage forms of one active ingredient combined with the dosage forms of another active ingredient. Unless the context makes clear otherwise, "combination therapy" may include different routes of administration for each active ingredient. Dosage forms, routes of administration and pharmaceutical compositions include, but are not limited to, those described herein.
A "stereoisomer" refers to a compound made up of the same atoms bonded by the same bonds but having different three-dimensional structures, which are not interchangeable. The present invention contemplates various stereoisomers and mixtures thereof and includes "enantiomers", which refers to two stereoisomers whose molecules are nonsuperimposeable mirror images of one another.
A "tautomer" refers to a proton shift from one atom of a molecule to another atom of the same molecule. The present invention includes tautomers of any said compounds.
The chemical naming protocol and structure diagrams used herein for the DMT1 inhibitors are a modified form of the I.U.P.A.C. nomenclature system, using the ChemDraw Version 10 software program (CambridgeSoft®). Unless the context requires otherwise, throughout the specification and claims which follow, the transitional phrases "comprise" and variations thereof, such as, "comprises", "comprising", "comprising of and "comprised of are to be construed to be synonymous with "including", "containing" or "characterized by" and are to be construed to be inclusive and open-ended in that additional, unrecited elements or method steps are not excluded.
DMT1 Inhibitors of the Invention
Inhibition of iron influx in enterocytes may be achieved either by direct inhibition of DMT1 or indirectly by removing the driving force for electrogenic iron influx. Iron transport via DMT1 in enterocytes involves an inwardly rectifying cation influx (Xu et a/., PIoS Biology 2: 378-386, 2004). Thus, maintained cation influx requires a counter- current to maintain the driving force for cation entry. Blocking this counter-current will inhibit iron influx and such block can be used in isolation or in combination with direct inhibition of DMT1 , with a chelator or with a proton pump inhibitor. Most often the counter-current is provided by potassium efflux through an ion channel selectively permeable to potassium or to chloride influx through an ion channel selectively permeable to chloride.
Examples of DMT1 inhibitors utilized in the combination therapies of the invention are the compounds described in PCT Published Application No. WO 2008/109840; PCT Published Application No. WO 2008/115999; PCT Published Application No. WO 2008/118790; PCT Published Application No. WO 2008/121861 ; and PCT Published Application No. WO 2008/151288.
More specifically, DMT1 inhibitors utilized in the combination therapy of the invention are compounds of formula (I); as described in PCT Published Application No. WO 2008/109840:
Figure imgf000018_0001
wherein: n and m are each independently 0, 1 or 2;
R1 and R2 are each independently a direct bond, -C(R9)2-, -S-, -O-, -C(O)-, -N(R9)- or
Figure imgf000018_0002
R3 and R4 are different and are each independently selected from
-R11-S-C(=NR12)N(R12)R13, -R11-O-C(=NR12)N(R12)R13, -R11-C(=NR12)N(R12)R13, or -R11-N(R9)-C(=NR12)N(R12)R13; or R3 and R4 are the same and are selected from -R11-S-C(=NR12)N(R12)R13,
-R11-O-C(=NR12)N(R12)R13, -R11-C(=NR12)N(R12)R13, or
-R11-N(R9)-C(=NR12)N(R12)R13; R5 and R6 are different and are each independently selected from hydrogen, alkyl, halo, haloalkyl, -R11-CN, -R11-NO2, -R11-N(R14)2, -R11-C(O)OR14,
-R11-C(O)N(R14)2, -R11-S-C(=NR12)N(R12)R13, -R11-O-C(=NR12)N(R12)R13, -R11-C(=NR12)N(R12)R13, -R11-N(R9)-C(=NR12)N(R12)R13, -N(R14)S(O)tR15, -S(O)tOR15, -S(O)pR14, or -S(O)tN(R14)2, wherein each t is independently 1 or 2 and each p is 0, 1 or 2; or R5 and R6 are the same and are selected from hydrogen, alkyl, halo, haloalkyl, -R11-CN, -R11-NO2, -R11-N(R14)2> -R11-C(O)OR14, -R11-C(O)N(R14)2,
-R11-S-C(=NR12)N(R12)R13, -R11-O-C(=NR12)N(R12)R13, -R11-C(=NR12)N(R12)R13, -R11-N(R9)-C(=NR12)N(R12)R13, -N(R14)S(O)tR15, -S(O)1OR15, -S(O)PR14, or -S(O)tN(R14)2, wherein each t is independently 1 or 2 and each p is 0, 1 or 2; each R7 and R8 is independently selected from alkyl, alkenyl, alkynyl, halo, haloalkyl, haloalkenyl, haloalkoxy, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted aralkenyl, optionally substituted heterocyclyl, optionally substituted heterocyclylalkyl, optionally substituted heteroaryl, optionally substituted heteroarylalkyl, -R11-CN, -R11-NO2, -R11-OR9, -R5-OS(O)2R15, -R11-N(R14)2, -R11-S(O)PR14, -R11-C(O)R14, -R11-C(S)R15, -R11-C(O)OR14,
-R11-OC(O)R14, -R11-C(S)OR14, -R11-C(O)N(R14)2, -R11-C(S)N(R14)2, -N=C(R15)2, -R11-N(R14)C(O)R15, -R11-N(R14)C(S)R15, -R11-N(R14)C(O)OR14, -R11-N(R14)C(S)OR14, -R11-N(R14)C(O)N(R14)2, -R11-N(R14)C(S)N(R14)2, -R11-N(R14)S(O)tR14, -R11-N(R14)S(O),N(R14)2, -R11-S(O)»N(R14)2> -R11-N(R14)C(=NR14)N(R14)2, or -R11-N(R14)C(N=C(R14)2)N(R14)2, wherein each p is independently 0, 1 , or 2 and each t is independently 1 or 2; each R9 is hydrogen, alkyl, alkenyl, alkynyl, haloalkyl, alkoxyalkyl, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heterocyclyl, optionally substituted heterocyclylalkyl, optionally substituted heteroaryl or optionally substituted heteroarylalkyl;
R10 is -C(R9)2-, -S-, -O- or -N(R9)-; each R11 is independently a direct bond or a straight or branched alkylene chain; each R12 and R13 is independently hydrogen, alkyl, or -OR9; each R14 is independently hydrogen, alkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heteroaryl or optionally substituted heteroaryl; and each R15 is alky!; as stereoisomers, enantiomers, tautomers thereof or mixtures thereof; or pharmaceutically acceptable salts thereof. Alternatively, DMT1 inhibitors utilized in the combination therapy of the invention are compounds of formula (II), as also described in PCT Published Application No. WO 2008/109840:
Figure imgf000020_0001
wherein: q and r are each independently 0, 1 or 2;
R16 and R17 are each independently =C(R24)- or =N-;
R i18 and R ,19 are different and are each independently selected from
-R25-S-C(=NR26)N(R26)R27, -R25-O-C(=NR26)N(R26)R27, -R25-C(=NR26)N(R26)R2
Figure imgf000020_0002
or R 5I18B and R 51'9a are the same and are selected from
Figure imgf000020_0003
-R25-O-C(=NR26)N(R26)R27, -R25-C(=NR26)N(R26)R27 or
Figure imgf000020_0004
R20 and R21 are different and are each independently selected from hydrogen, alkyl, halo, haloalkyl, -R25-CN, -R25-NO2, -R25-N(R28)2, -R25-C(O)OR28, -R25-C(O)N(R28)2, -R25-S-C(=NR26)N(R26)R27, -R25-O-C(=NR26)N(R26)R27, -R25-C(=NR26)N(R26)R27, -R25-N(R9)-C(=NR26)N(R26)R27, -N(R28)S(O),R29,
-S(O)tOR , -S(O)PR , or -S(O)1N(R^)2, wherein each t is independently 1 or 2 and each p is 0, 1 or 2; or R20 and R21 are the same and are selected from hydrogen, alkyl, halo, haloalkyl, -R25-CN, -R25-NO2, -R25-N(R28)2, -R25-C(O)OR28, -R25-C(O)N(R28)2, -R25-S-C(=NR26)N(R26)R27, -R25-O-C(=NR26)N(R26)R27, -R25-C(=NR26)N(R26)R27, -R25-N(R9)-C(=NR26)N(R26)R27, -N(R28)S(O)tR29, -S(O),OR29, -S(O)PR28, or -S(O)tN(R28)2> wherein each t is independently 1 or 2 and each p is 0, 1 or 2; each R22 and R23 is independently selected from alkyl, alkenyl, alkynyl, halo, haloalkyl, haloalkeny!, haloalkoxy, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted aralkenyl, optionally substituted heterocyclyl, optionally substituted heterocyclylalkyl, optionally substituted heteroaryl, optionally substituted heteroarylalkyl, -R25-CN, -R25-NO2, -R25-OR24, -R25-OS(O)2R29, -R25-N(R28)2, -R25-S(O)PR28, -R25-C(O)R28, -R25-C(S)R29, -R25-C(O)OR28, -R25-OC(O)R28, -R25-C(S)OR28, -R25-C(O)N(R28)2, -R25-C(S)N(R28)2> -N=C(R29)2, -R25-N(R28)C(O)R29, -R25-N(R28)C(S)R29, -R25-N(R28)C(O)OR28, -R25-N(R28)C(S)OR28, -R25-N(R28)C(O)N(R28)2, -R25-N(R28)C(S)N(R28)2,
-R25-N(R28)S(O)tR28, -R25-N(R28)S(O)tN(R28)2, -R25-S(O)tN(R28)2, -R25-N(R28)C(=NR28)N(R28)2> or -R25-N(R28)C(N=C(R28)2)N(R28)2, wherein each p is independently 0, 1 , or 2 and each t is independently 1 or 2; each R24 is hydrogen, alkyl, alkenyl, alkynyl, haloalkyl, alkoxyalkyl, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heterocyclyl, optionally substituted heterocyclylalkyl, optionally substituted heteroaryl or optionally substituted heteroarylalkyl; each R25 is independently a direct bond or a straight or branched alkylene chain; each R26 and R27 is independently hydrogen, alkyl or -OR24; each R28 is independently hydrogen, alkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heteroaryl or optionally substituted heteroaryl; and each R29 is alkyl; as stereoisomers, enantiomers, tautomers thereof or mixtures thereof; or pharmaceutically acceptable salts thereof.
Alternatively, DMT1 inhibitors utilized in the combination therapy of the invention are compounds of formula (I), as described in PCT Published Application No.
WO 2008/115999:
(R2)m— ( A )-R1{ B V(R3)π (I)
wherein: n and m are each independently 1 , 2, 3, 4, 5, 6 or 7;
Figure imgf000021_0001
and are each independently aryl or heteroaryl;
R1 is a direct bond, -O-, -S(O)P- (wherein p is 0, 1 or 2), -C(R4)2-, -C(O)- or -N(R4)-; at least one R2 and at least one R3 is independently selected from of
-R6-S-C(=NR4)N(R4)R5, -R6-S-C(=NR4)N(R4)N(R4)R5, -R6-O-C(=NR4)N(R4)R5, -R6-C(O)-N=C[N(R4)(R5)]N(R4)R5, -R6-C(=NR4)N(R4)R5, -R6-C(=NCN)N(R4)R5 or -R6-N(R7)C(=NR4)N(R4)R5; and the other R2's and R3's, if present, are each independently selected from alkyl, halo, haloalkyl, -R6-OR7,-R6-CN, -R6-NO2, -R6-N(R8)2, -R6-C(O)OR8, -R6-C(O)N(R8)2, -N(R8)S(O)tR9, -S(O)1OR9, -S(O)PR8, -S(O)tN(R8)2,
-R6-S-C(=NR4)N(R4)R5, -R6-O-C(=NR4)N(R4)R5, -R6-C(=NR4)N(R4)R5, and
-R6-N(R7)-C(=NR4)N(R4)R5, wherein each t is independently 1 or 2 and each p is 0, 1 or 2; each R4 and R5 is independently hydrogen, alkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted heteroaryl, or optionally substituted heteroaralkyl; each R6 is independently a direct bond or a straight or branched alkylene chain; each R7 is hydrogen, alkyl, alkenyl, alkynyl, haloalkyl, alkoxyalkyl, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heterocyclyl, optionally substituted heterocyclylalkyl, optionally substituted heteroaryl or optionally substituted heteroarylalkyl; each R8 is independently hydrogen or alkyl; and each R9 is alkyl; as stereoisomers, enantiomers, tautomers thereof or mixtures thereof; or pharmaceutically acceptable salts thereof.
Alternatively, DMT1 inhibitors utilized in the combination therapy of the invention are compounds of formula (I), as described in PCT Published Application No. WO 2008/118790:
Figure imgf000022_0001
wherein:
Figure imgf000022_0002
is a fused aryl ring or a fused heteroaryl ring; m is O, 1 , 2, 3 or 4;
R1 is an optionally substituted aryl or an optionally substituted heteroaryl;
R2 is -OR5, -OC(O)R5, or -S(O)PR5 (wherein p is 0, 1 or 2); each R3 is independently selected from alkyl, alkenyl, alkynyl, halo, haloalkyl, haloalkenyl, haloalkynyl, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted cycloalkylalkenyl, optionally substituted cycloalkylalkynyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted aralkenyl, optionally substituted aralkynyl, optionally substituted heterocyclyl, optionally substituted heterocyclylalkyl, optionally substituted heterocyclylalkenyl, optionally substituted heterocyclylalkynyl, optionally substituted heteroaryl, optionally substituted heteroarylalkyl, optionally substituted heteroarylalkenyl, optionally substituted heteroarylalkynyl, -R8-OR6, -R8-CN, -R8-OC(O)-R6, -R8-C(O)R6, -R8-C(O)OR6, -R8-C(O)N(R6)2, -R8-NO2, -R8-N(R6)2, -R8-N(R6)C(O)OR7, -R8-N(R6)C(O)R7, -R8-N(R6)S(O),R7 (wherein t is 1 to 2), -R8-S(O)tOR7 (wherein t is 1 to 2), -R8-S(O)PR7 (wherein p is 0, 1 or 2), or -R8-S(O)tN(R6)2 (wherein t is 1 to 2);
R4 is -[C(R6)2]n- wherein n is 2 or 3;
R5 is hydrogen or alkyl; each R6 is independently hydrogen, alkyl, alkenyl, alkynyl, haloalkyl, alkoxyalkyl, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heterocyclyl, optionally substituted heterocyclylalkyl, optionally substituted heteroaryl or optionally substituted heteroarylalkyl; each R7 is independently alkyl, alkenyl, alkynyl, haloalkyl, alkoxyalkyl, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heterocyclyl, optionally substituted heterocyclylalkyl, optionally substituted heteroaryl or optionally substituted heteroarylalkyl; and each R8 is a direct bond or a straight or branched alkylene chain; as stereoisomers, enantiomers, tautomers thereof or mixtures thereof; or pharmaceutically acceptable salts thereof.
Alternatively, DMT1 inhibitors utilized in the combination therapy of the invention are compounds of formula (I), as described in PCT Published Application No.
WO 2008/121861 :
Figure imgf000024_0001
wherein:
R1 is hydrogen, hydroxyalkyl, optionally substituted aryl, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted aralkyl, optionally substituted heteroaryl or optionally substituted heteroarylalkyl;
R2 is hydrogen, alkyl, alkenyl, alkynyl, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted cycloalkylalkenyl, optionally substituted cycloalkylalkynyl, optionally substituted heterocyclyl, optionally substituted heterocyclylalkyl, optionally substituted heterocyclylalkenyl or optionally substituted heterocyclylalkynyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted aralkenyl, optionally substituted aralkynyl, optionally substituted heteroaryl, optionally substituted heteroarylalkyl, optionally substituted heteroarylalkenyl or optionally substituted heteroarylalkynyl; R3 is optionally substituted aryl or optionally substituted heteroaryl;
R4 is alkyl, halo, haloalkyl, -OR5, -OC(O)R5, -OS(O)2R5, -S(O)PR5 (wherein p is O, 1 or 2), or -N(R5)2; and, each R5 is independently hydrogen, alkyl or haloalkyl; as stereoisomers, enantiomers, tautomers thereof or mixtures thereof; or pharmaceutically acceptable salts thereof.
Alternatively, DMT1 inhibitors utilized in the combination therapy of the invention are compounds of formula (I), as also described in PCT Published
Publication No. WO 2008/121861 :
Figure imgf000024_0002
wherein;
R7 is hydrogen, hydroxyalkyl, optionally substituted aryl, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted aralkyl, optionally substituted heteroaryl or optionally substituted heteroarylalkyl;
R8 is hydrogen, alkyl, alkenyl, alkynyl, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted cycloalkylalkenyl, optionally substituted cycloalkylalkynyl, optionally substituted heterocyclyl, optionally substituted heterocyclylalkyl, optionally substituted heterocyclylalkenyl or optionally substituted heterocyclylalkynyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted aralkenyl, optionally substituted aralkynyl, optionally substituted heteroaryl, optionally substituted heteroarylalkyl, optionally substituted heteroarylalkeny! or optionally substituted heteroarylalkynyl;
R9 is hydrogen, optionally substituted aryl, optionally substituted aralkyl, -C(O)OR11 or -C(O)N(R11)2;
R10 is hydrogen, alkenyl, alkynyl, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted cycloalkylalkenyl, optionally substituted cycloalkylalkynyl, optionally substituted heterocyclyl, optionally substituted heterocyclylalkyl, optionally substituted heterocyclylalkenyl or optionally substituted heterocyclylalkynyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted aralkenyl, optionally substituted aralkynyl, optionally substituted heteroaryl, optionally substituted heteroarylalkyl, optionally substituted heteroarylalkenyl or optionally substituted heteroarylalkynyl; and each R11 is independently hydrogen, alkyl, haloalkyl, optionally substituted aryl, or optionally substituted heteroaryl; as stereoisomers, enantiomers, tautomers thereof or mixtures thereof; or pharmaceutically acceptable salts thereof.
Alternatively, DMT1 inhibitors utilized in the combination therapy of the invention are compounds of formula (I), as also described in PCT Published
Publication No. WO 2008/121861 :
Figure imgf000025_0001
wherein:
R12 is hydrogen, hydroxyalkyl, optionally substituted aryl, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted aralkyl, optionally substituted heteroaryl or optionally substituted heteroarylalkyl; R13 is hydrogen, alkyl, alkenyl, alkynyl, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted cycloalkylalkenyl, optionally substituted cycloalkylalkynyl, optionally substituted heterocyclyl, optionally substituted heterocyclylalkyl, optionally substituted heterocyclylalkenyl or optionally substituted heterocyclylalkynyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted aralkenyl, optionally substituted aralkynyl, optionally substituted heteroaryl, optionally substituted heteroarylalkyl, optionally substituted heteroarylalkenyl or optionally substituted heteroarylalkynyl;
R14a and R14b are each independently selected from hydrogen, alkyl, halolalkyl, -C(O)OR15 or -C(O)N(R15)2; or R14a and R14b together form =C(R15)R16; each R15 is independently hydrogen, alkyl, alkenyl, alkynyl, haloalkyl, alkoxyalkyl, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heterocyclyl, optionally substituted heterocyclylalkyl, optionally substituted heteroaryl or optionally substituted heteroarylalkyl; and
R16 is optionally substituted aryl, optionally substituted aralkyl, optionally substituted heteroaryl or optionally substituted heteroarylalkyl; as stereoisomers, enantiomers, tautomers thereof or mixtures thereof; or pharmaceutically acceptable salts thereof.
Alternatively, DMT1 inhibitors utilized in the combination therapy of the invention are compounds of formula (I), as also described in PCT Published
Application No. WO 2008/121861 :
Figure imgf000026_0001
wherein: R17 is hydrogen, hydroxyalkyl, optionally substituted aryl, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted aralkyl, optionally substituted heteroaryl or optionally substituted heteroarylalkyl;
R18 is hydrogen, alkyl, alkenyl, alkynyl, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted cycloalkylalkenyl, optionally substituted cycloalkylalkynyl, optionally substituted heterocyclyl, optionally substituted heterocyclylalkyl, optionally substituted heterocyclylalkenyl or optionally substituted heterocyclylalkynyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted aralkenyl, optionally substituted aralkynyl, optionally substituted heteroaryl, optionally substituted heteroarylalkyl, optionally substituted heteroarylalkenyl or optionally substituted heteroarylalkynyl;
R19 is hydrogen, alkyl, halolalkyl, -C(O)OR21 or -C(O)N(R21)2;
R20a and R20b are each independently selected from hydrogen, alkyl, halolalkyl, -C(O)OR21 or -C(O)N(R21)2; or R20a and R20b together form =C(R21)R22; each R21 is independently hydrogen, alkyl, alkenyl, alkynyl, haloalkyl, alkoxyalkyl, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heterocyclyl, optionally substituted heterocyclylalkyl, optionally substituted heteroaryl or optionally substituted heteroarylalkyl; and
R22 is optionally substituted aryl, optionally substituted aralkyl, optionally substituted heteroaryl or optionally substituted heteroarylalkyl; as stereoisomers, enantiomers, tautomers thereof or mixtures thereof; or pharmaceutically acceptable salts thereof.
Alternatively, DMT1 inhibitors utilized in the combination therapy of the invention are compounds of formula (I), as described in PCT Published Application No.
WO 2008/151288:
Figure imgf000027_0001
wherein: m is O, 1 , 2, 3, or 4;
Figure imgf000028_0001
is aryl or heteroaryl;
R1 and R2 are each independently selected from -R6-S-C(=NR4)N(R4)R5, -R6-C(O)-S-C(=NR4)N(R4)R5, -R6-S-C(=NR4)N(R4)N(R4)R5, -R6-O-C(=NR4)N(R4)R5, -R6-C(O)-N=C[N(R4)(R5)]N(R4)R5, -R6-C(=NR4)N(R4)R5, -R6-C(=NCN)N(R4)R5, -R6-N(R7)C(=NCN)N(R4)R5 or -R6-N(R7)C(=NR4)N(R4)R5; each R3 is independently selected from hydrogen, alkyl, halo, haloalkyl, optionally substituted aryl, -R6-OR7, -R6-CN, -R6-NO2, -R6-N(R8)2, -R6-C(O)OR8, -R6-C(O)N(R8)2> -N(R8)S(O)tR9, -S(O)1OR9, -S(O)PR8, -S(O)tN(R8)2, -R6-S-C(=NR4)N(R4)R5, -R6-O-C(=NR4)N(R4)R5, -R6-C(=NR4)N(R4)R5, or -R6-N(R7)-C(=NR4)N(R4)R5, wherein each t is independently 1 or 2 and each p is 0, 1 or 2; each R4 and R5 is independently hydrogen, alkyl, or -OR7; each R6 is independently a direct bond or a straight or branched alkylene chain; each R7 is hydrogen, alkyl, alkenyl, alkynyl, haloalkyl, alkoxyalkyl, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heterocyclyl, optionally substituted heterocyclylalkyl, optionally substituted heteroaryl or optionally substituted heteroarylalkyl; each R8 is independently hydrogen or alkyl; and each R9 is alkyl; as stereoisomers, enantiomers, tautomers thereof or mixtures thereof; or pharmaceutically acceptable salt thereof.
The following compounds, or their pharmaceutically acceptable salts, which are disclosed in the aforementioned applications, are DMT1 inhibitors and are therefore useful in the practice of the invention:
(2,4,6-trimethyl-1 ,3-phenylene)bis(methylene) dicarbamimidothioate; dibenzo[b,c/]thiophene-4,6-diylbis(methylene) dicarbamimidothioate;
(2-fluorodibenzo[jb,d]thiophene-4,6-diyl)bis(methylene) dicarbamimidothioate;
(3,7-dibromodibenzo[/),c(]thiophene-4,6-diyl)bis(methylene) dicarbamimidothioate; (2-chloro-8-fluorodibenzo[jb,c(]thiophene-4,6-diyl)bis(methylene) dicarbamimidothioate;
(3-bromodibenzo[6,c/|thiophene-4,6-diyl)bis(methylene) dicarbamimidothioate; dibenzo[b,c/]thiophene-1 ,9-diylbis(methylene) dicarbamimidothioate;
(9-oxo-9/-/-xanthene-4,5-diyl)bis(methylene) dicarbamimidothioate; dibenzo[b,c0furan-4,6-diylbis(methylene) dicarbamimidothioate;
(3,7-dimethyldibenzo[ib,c/]furan-4,6-diyl)bis(methylene) dicarbamimidothioate;
(3,7-dichloroldibenzo[b,c/]furan-416-diyi)bis(methylene) dicarbamimidothioate;
(3,7-dibromodibenzo[ifc),c/lfuran-4,6-diyl)bis(methylene) dicarbamimidothioate; (2-fluorodibenzo[)b,c/]furan-4,6-diyl)bis(methylene) dicarbamimidothioate;
(2,8-dibromodibenzo[jb,d]furan-4,6-diyl)bis(methylene) dicarbamimidothioate; biphenylene-1 ,8-diylbis(methylene) dicarbamimidothioate;
(3,6-difluorobiphenylene-1 ,8-diyl)bis(methylene) dicarbamimidothioate; biphenylene-1 , 4, 5, δ-tetrayltetrakis(methylene) tetracarbamimidothioate; 2-(8-carbamimidoylsulfanylmethyl-9-oxo-9H-fluoren-1-ylmethyl)-isothiourea;
(9-oxo-9H-fluorene-4,5-diyl)bis(methylene) dicarbamimidothioate;
2-(2,7-di-tert-butyl-5-carbamimidoylsulfanylmethyl-9,9-dimethyl-9/-/-xanthen-4- ylmethyl)-isothiourea; phenoxathiine-4,6-diylbis(methylene) dicarbamimidothioate; (5,7-dihydrodibenzo[c,e]thiepine-1 , 11 -diyl)bis(methylene) dicarbamimidothioate;
2-(2'-carbamimidoylsulfanylmethyl-biphenyl-2-ylmethyl)-isothiourea;
(6,6'-dimethylbiphenyl-2,2'-diyl)bis(methylene) dicarbamimidothioate dihydrobromide; biphenyl-2,2',6,6'-tetrayltetrakis(methylene) tetracarbamimidothioate; dimethyl 6,6'-bis(carbamimidoylthiomethyl)biphenyl-2,2'-dicarboxylate; 2-[2-(2-carbamimidoylsulfanylmethyl-phenoxy)-benzyl]-isothiourea;
2-(1-{2-[2-(1-carbamimidoylsulfanyl-ethyl)-phenoxy]-phenyl}-ethyl)-isothiourea;
2-[2-({[amino(imino)methyl]thio}methyl)-4-fluorophenoxy]-5-nitrobenzyl imidothiocarbamate;
2-[2-({[amino(imino)methyl]thio}methyl)-5-chlorophenoxy]-4-nitrobenzyl imidothiocarbamate;
2-[2-({[amino(imino)methyl]thio}methyl)-5-chlorophenoxy]-5-fluorobenzyl imidothiocarbamate;
2-[2-(2-carbamimidoylsulfanylmethyl-3-chlorophenoxy)-5-fluorobenzyl]isothiourea;
2-[2-(2-carbamimidoylsulfanylmethyl-4-chlorophenoxy)-5-fluorobenzyI]isothiourea; 2-[2-({[amino(imino)methyl]thio}methyl)-4-chlorophenoxy]benzyl imidothiocarbamate;
2-[2-({[amino(imino)methyl]thio}methyl)-3-chlorophenoxy]benzyl imidothiocarbamate;
2-[2-({[amino(imino)methyl]thio}methyl)-5-chlorophenoxy]benzyl imidothiocarbamate;
2-[2-({[amino(imino)methyl]thio}methyl)-5-chlorophenoxy]-5-nitrobenzyl imidothiocarbamate; 2-[2-({[amino(imino)methyl]thio}methyl)-4-chlorophenoxy]-5-nitrobenzyl imidothiocarbamate;
2-[2-(2-carbamimidoylsulfanylmethyl-5-fluorophenoxy)-5-fluorobenzyl]isothiourea; 2-[2-(2-carbamimidoylsulfanylmethyl-4-fluorophenoxy)-5-fluorobenzyl]isothiourea; 2-[2-({[amino(imino)nnethyl]thio}methyl)-4-fluorophenoxy]benzyl imidothiocarbamate; 2-[2-(2-carbamimidoylsulfanylmethyl-phenylsulfanyl)-benzyl]-isothiourea;
2-[2-(2-carbamimidoylsulfanylmethyl-4-fluoro-phenylsulfanyl)-5-fluoro-benzyl]- isothiourea;
2-[2-(2-carbamimidoylsulfanylmethyl-4-fluoro-phenylsulfanyl)-benzyl]-isothiourea; 2-[2-(2-carbamimidoylsulfanylmethyl-5-methyl-phenylsulfanyl)-benzyl]-isothiourea; 2-[2-(2-carbamimidoylsulfanylmethyl-4-methoxy-phenylsulfanyl)-benzyl]-isothiourea; 2-[2-(2-carbamimidoylsulfanylmethyl-5-methyl-phenylsulfanyl)-5-fluoro-benzyl]- isothiourea; 2-[2-(2-carbamimidoylsulfanylmethyl-4-methoxy-phenylsulfanyl)-5-fluoro-benzyl]- isothiourea; 2-[2-(2-carbamimidoylsulfanylmethyl-5-chloro-phenylsulfanyl)-5-fluoro-benzyl]- isothiourea;
2-[2-(2-carbamimidoylsulfanylmethyl-6-methylphenylsulfanyl)-benzyl]isothiourea; 2-[2-(2-carbamimidoyIsulfanylmethyl-4,5-difluorophenylsulfanyl)-benzyl]isothiourea; 2-[2-(2-carbamimidoylsulfanylmethyl-4-methyl-phenylsulfanyl)-benzyl]-isothiourea; 2-[2-(2-carbamimidoylsulfanylmethyl-5-chloro-phenylsulfanyl)-benzyl]-isothiourea; 2-[2-(2-carbamimidoylsulfanylmethyl-5-fluoro-phenylsulfanyl)-benzyl]-isothiourea; 2-[2-(2-carbamimidoylsulfanylmethyl-5-fluoro-phenylsulfanyl)-5-fluorobenzyl]- isothiourea;
2-[2-(2-carbamimidoylsulfanylmethyl-4,5-difluorophenylsulfanyl)-5- fluorobenzyφsothiourea;
2-{[2-({[amino(imino)methyl]thio}methyl)-4-fluorophenyl]thio}-3-nitrobenzyl imidothiocarbamate; 2-{[2-({[amino(imino)methyl]thio}methyl)-4-fluorophenyl]thio}-5-nitrobenzyI imidothiocarbamate; 2-[2-(2-carbamimidoylsulfanylmethyl-4-fluorophenylsulfanyl)-5-
(trifluoromethyl)benzyl]isothiourea;
2-[2-(2-carbamimidoylsulfanylmethylphenylsulfanyl)-5-aminobenzyl]-isothiourea; 2-[2-(2-carbamimidoylsulfanylmethyl-4-chlorophenylsulfanyl)-5- fluorobenzyφsothiourea; 2-[2-(2-carbamimidoylsulfanyImethylphenylsulfanyl)-5-ethylaminobenzyl]isothiourea; 2-[2-(2-carbamimidoylsulfanylmethyl-4-chlorophenylsulfanyl)-5- chlorobenzyφsothiourea;
2-[2-(2-carbamimidoylsulfanylethyl-4-fluorophenylsulfanyl)-5-fluorobenzyl]isothiourea;
2-[2-(2-carbamimidoy!sulfanylethyl-4-chlorophenylsulfanyl)-5-fluorobenzyl]isothiourea; 2-[2-(2-carbamimidoylsulfanylethylphenylsulfanyl)benzyl]isothiourea;
2-[2-(2-carbamimidoylsulfanylmethylphenylsulfanyl)-5-chlorobenzyl]-isothiourea;
2-[2-(2-carbamimidoylsulfanylmethylphenylsulfanyl)-5- (trifluoromethyl)benzyl]isothiourea;
2-[2-(2-methylcarbamimidoylsulfanylmethylphenylsulfanyl)benzyl]-methylisothiourea; 2-[2-(2-carbamimidoylsulfanylmethyl-4-fluorophenylsulfanyl)-5- (methylsulfonyl)benzyl]isothiourea;
2-({2-({[amino(imino)methyl]thio}methy!)-4-[(dimethylamino)sulfonyl]phenyl}thio)-5- fluorobenzyl imidothiocarbamate;
2-[2-(2-carbamimidoylsulfanyImethylphenylsulfanyl)benzyl]-isothiourea; 2-{[2-({[amino(imino)methyl]thio}methyl)-4-fluorophenyl]thio}-4-(methylsulfonyl)benzyl imidothiocarbamate;
2-{[2-({[amino(imino)methyl]thio}methyl)-4-chlorophenyl]thio}-5-cyanobenzyl imidothiocarbamate;
2-[2-(2-carbamimidoylsulfanylmethylphenylsulfonyl)benzyl]-isothiourea; 2-[2-(2-carbamimidoylsuifanylmethyl-4-fluorophenylsulfonyl)-5-fluorobenzyl]isothiourea;
2-(6-((aminoamidino)thiomethyl)phenyl)thio-1-((aminoamidino)thiomethyl)benzene;
2-(6-(2-amidinoethyl)phenyl)thio-1-(2-amidinoethyl)benzene;
2-(6-(amidinomethyl)phenyl)thio-1-(amidinomethyl)benzene;
2-(6-(3-amidinopropyl)phenyl)thio-1-(3-amidinopropyl)benzene; 2-(6-((cyanoamidino)methyl)phenyl)thio-1-((cyanoamidino)methyl)benzene;
2-(6-(2-(cyanoamidino)ethyl)phenyl)thio-1-(2-(cyanoamidino)ethyl)benzene;
2-(6-(3-(cyanoamidino)propyl)phenyl)thio-1-(3-(cyanoamidino)propyl)benzene;
1-(2-(2-(guanidinomethyl)phenylthio)benzyl)guanidine;
2,2'-thiobis(Λ/-(diaminomethylene)benzamide); 2-(6-(amidinothiomethyl)phenyl)carbonyl-1-(amidinothiomethyl)benzene;
2-(6-(amidinothiomethyl)phenyl)carbonyl-1-(amidinothiomethyl)-5-fluorobenzene;
2-(6-(amidinothiomethyl)phenyl)carbonyl-1-(amidinothiomethyl)-4-fluorobenzene;
2-(6-(amidinothiomethyl)phenyl)carbonyl-1-(amidinothiomethyl)-4-chlorobenzene;
2-(6-(amidinothiomethyl)phenyl)carbonyl-1-(amidinothiomethyl)-5-chlorobenzene; 4,4-diisothiourea benzophenone; 2,2-(methylazanediyl)bis(2,1-phenylene)bis(methylene)dicarbamimidothioate;
2-[2-(1-carbamimidoylsulfanylmethyl-naphthalen-2-ylsulfanyl)-benzyl]-isothiourea;
2-[2-(1-carbamimidoylsulfanylmethylnaphthalen-2-ylsulfanyl)-5-fluorobenzyl]- isothiourea; (2-{[2-({[amino(imino)methyl]thio}methyl)-4-fluorophenyl]thio}-3-thienyl)methyl imidothiocarbamate;
(4-{[2-({[amino(imino)methyl]thio}methyl)-4-fluorophenyl]thio}-3-thienyl)methyl imidothiocarbamate;
2-pyridin-2-yl-4,5-dihydro-2/-/-benzo[e]indazol-1-ol; 7-bromo-2-pyridin-2-yl-4,5-dihydro-2/-/-benzo[e]indazol-1 -ol;
2-pyridin-2-yl-2,4,5,6-tetrahydro-2,3-diaza-benzo[e]azulen-1-ol;
1-hydroxy-2-pyridin-2-yl-4,5-dihydro-2/-/-benzo[e]indazole-7-carbonitrile;
7-phenyl-2-pyridin-2-yl-4,5-dihydro-2/-/-benzo[e]indazol-1-ol;
7-benzo[1 ,3]dioxol-5-yl-2-pyridin-2-yl-4,5-dihydro-2/-/-benzo[e]indazol-1-ol; 4-(1-hydroxy-2-pyridin-2-yl-4,5-dihydro-2H-benzo[e]indazol-7-yl)-benzonitrile;
2-pyridin-2-yl-7-p-tolyl-4,5-dihydro-2/-/-benzo[e]indazol-1-ol;
7-(4-methoxyphenyl)-2-pyridin-2-yl-4,5-dihydro-2/-/-benzo[e]indazol-1-ol;
2-(1-hydroxy-4,5-dihydro-2/-/-benzo[e]indazol-2-yl)pyridine 1 -oxide;
7-methoxy-2-pyridin-2-yl-4,5-dihydro-2/-/-benzo[e]indazol-1-ol; 8-methoxy-2-pyridin-2-yl-4,5-dihydro-2H-benzo[e]indazol-1-ol;
2-(5-(trifluoromethyl)pyridin-2-yl)-4,5-dihydro-2/-/-benzo[e]indazol-1-ol;
2-(4-(trifluoromethyl)pyridin-2-yl)-4,5-dihydro-2/-/-benzo[e]indazol-1-ol;
2-(5-nitropyridin-2-yl)-4,5-dihydro-2/-/-benzo[e]indazol-1-ol;
2-(3-(trifluoromethyl)pyridin-2-yl)-4,5-dihydro-2/-/-benzo[e]indazol-1-ol; 2-(5-methylpyridin-2-yl)-4,5-dihydro-2/-/-benzo[e]indazol-1-ol;
2-(benzo[c/]thiazol-2-yl)-4,5-dihydro-2/-/-benzo[e]indazol-1-ol;
2-(6-methoxybenzo[c/]thiazol-2-yl)-4,5-dihydro-2/-/-benzo[e]indazol-1-ol;
2-(6-fluorobenzo[c/]thiazol-2-yl)-4,5-dihydro-2H-benzo[e]indazol-1-ol;
2-(benzo[cf]thiazol-2-yl)-7-methoxy-4,5-dihydro-2/-/-benzo[e]indazol-1-ol; 2-(6-methylbenzo[c(]thiazol-2-yl)-4,5-dihydro-2H-benzo[e]indazol-1-ol;
2-(4-methylbenzo[c/]thiazol-2-yl)-4,5-dihydro-2H-benzo[e]indazol-1-ol;
2-(1-hydroxy-4,5-dihydro-2/-/-benzo[e]indazol-2-yl)benzo[c/]thiazole-6-carboxylic acid;
2-(1f7-benzo[c/]imidazol-2-yl)-4,5-dihydro-2/-/-benzo[e]indazol-1-ol;
2-(4-tert-butylthiazol-2-yl)-4,5'dihydro-2/-/-benzo[e]indazol-1-ol hydrochloride; 1 -(2-bromophenyl)-3-methyl-4-phenyl-1 H-pyrazol-5-amine; 3-methyl-1-(pyridin-2-yl)-4-(4-(trifluoromethyl)phenyl)-1/-/-pyrazol-5-ol;
3-methyl-4-phenyl-1 -(pyridin-2-yl)-1 /-/-pyrazol-5-amine;
2-(3,5-dimethyl-4-phenyl-1 H-pyrazol-1 -yl)pyridine;
3-methyl-4-phenyl-1 -(pyridin-2-yl)-1 H-pyrazol-5-yl trifluoromethanesulfonate; 3-methyl-1 -(pyridin-2-yl)-4-(3-(trifluoromethyl)phenyl)-1 H-pyrazol-5-ol;
4-(5-hydroxy-3-methyl-1-(pyridin-2-yl)-1H-pyrazol-4-yl)benzoic acid;
4-(5-hydroxy-3-methyl-1-(pyridin-2-yl)-1H-pyrazol-4-yl)-Λ/-(6-hydroxyhexyl)benzamide;
3-methyl-4-(4-(methylsulfonyl)phenyl)-1-(pyridin-2-yl)-1 H-pyrazol-5-ol;
4-(4-(hydroxymethyl)phenyl)-3-methyl-1-(pyridin-2-yl)-1 H-pyrazol-5-ol; 3-methyl-4-phenyl-1 -(pyrimidin-2-yl)-1 H-pyrazol-5-ol;
6-(5-hydroxy-3-methyl-4-phenylpyrazol-1-yl)-pyridazin-3-ol;
3-methyl-4-phenyl-1 -(thiazol-2-yl)-1 H-pyrazol-5-ol;
1 -(benzo[c/]thiazol-2-yl)-3-methyl-4-phenyl-1 H-pyrazol-5-ol;
1-(1 H-benzo[c/]imidazol-2-yl)-3-methyl-4-phenyl-1 H-pyrazol-5-ol; 1 -(isoquinolin-1 -yl)-3-methyl-4-phenyl-1 H-pyrazol-5-ol;
1-(6-methoxybenzo[c/]thiazoI-2-yl)-3-methyl-4-phenyl-1 H-pyrazol-5-ol;
1-(6-fluorobenzo[c/]thiazol-2-yl)-3-methyl-4-phenyl-1 H-pyrazol-5-ol;
3-methyl-1-(5-nitrothiazol-2-yl)-4-phenyl-1 H-pyrazol-5-ol;
3-methyl-4-(4-nitrophenyl)-1-(pyridin-2-yl)-1 H-pyrazol-5-ol; 4-(4-aminophenyl)-3-methyl-1-(pyridin-2-yl)-1 H-pyrazol-5-ol;
4-(4-methoxyphenyl)-3-methyl-1-(pyridin-2-yl)-1 H-pyrazol-5-ol;
4-(4-(diethylamino)phenyl)-3-methyl-1-(pyridin-2-yl)-1 H-pyrazol-5-ol;
4-(5-hydroxy-3-methyl-1-(pyridin-2-yl)-1H-pyrazol-4-yl)benzonitrile;
4-(4-fluorophenyl)-3-methyl-1 -(pyridin-2-yl)-1 H-pyrazol-5-ol; ethyl 4-(5-hydroxy-3-methyl-1 -(pyridin-2-yl)-1 H-pyrazol-4-yl)benzoate;
3-methyl-4-phenyl-1 -(pyridin-2-yl-methyl)-1 H-pyrazol-5-ol;
1 ,3-phenylenebis(methylene) dicarbamimidothioate;
(2-fluoro-1 ,3-phenylene)bis(methylene) dicarbamimidothioate;
1 ,3-phenylene dicarbamimidothioate; (5-methyl-1 ,3-phenylene)bis(methylene) dicarbamimidothioate;
(2,4,6-trimethylbenzene-1 ,3,5-triyl)tris(methylene) tricarbamimidothioate;
2-{1-[3-(1-carbamimidoylsulfanyl-1-methylethyl)phenyl]-1-methylethyl}isothiourea;
(2-cyano-1 ,3-phenylene)bis(methylene) dicarbamimidothioate;
(4,6-dimethyl-1 ,3-phenylene)bis(methylene) dicarbamimidothioate; diethyl 4,6-bis(carbamimidoylthiomethyl)isophthalate; (5-bromo-4,6-dimethyl-1 ,3-phenylene)bis(methylene) dicarbamimidothioate;
(2,4,5,6-tetramethyl-1 ,3-phenylene)bis(methylene) dicarbamimidothioate;
2-{1-[3-(1-carbamimidoylsulfanylethyl)-2,4,6-trimethylphenyl]ethyl}isothiourea;
(2-hydroxy-5-methyl-1 ,3-phenylene)bis(methylene) dicarbamimidothioate; 1 ,3-di[(methylamidino)thiomethyl]-2,4,6-trimethylbenzene;
(5-hydroxy-2,4,6-trimethyl-1 ,3-phenylene)bis(methylene) dicarbamimidothioate;
(2,4,5,6-tetrachloro-1 ,3-phenylene)bis(methylene) dicarbamimidothioate;
(2-methoxy-5-methyl-1 ,3-phenylene)bis(methylene) dicarbamimidothioate;
(2-methyl-1 ,3-phenylene)bis(methylene) dicarbamimidothioate; (4-methoxy-1 ,3-phenylene)bis(methylene) dicarbamimidothioate;
(5-methoxy-1 ,3-phenylene)bis(methylene) dicarbamimidothioate;
(4,6-dibromo-1 ,3-phenylene)bis(methylene) dicarbamimidothioate;
(4,6-diisopropyl-1 ,3-phenylene)bis(methylene) dicarbamimidothioate;
1 ,3-di[(2-cyano-3-methylguanidino)methyl]-2,4,6-trimethylbenzene; 2,2'-(1 ,3-phenylene)diacetimidamide;
Λ/-(3-guanidinomethyl-2,4,6-trimethylbenzyl)guanidine; pyridine-2,6-diylbis(methylene) dicarbamimidothioate;
(2,4,6-trimethylpyridine-3,5-diyl)bis(methylene) dicarbamimidothioate;
(1 ,2-phenylene)bis(methylene) dicarbamimidothioate; (3,4,5,6-tetramethyl-1 ,2-phenylene)bis(methylene) dicarbamimidothioate; naphthalene-1 ,2-diylbis(methylene) dicarbamimidothioate; naphthalene-1 ,8-diylbis(methylene) dicarbamimidothioate;
2-(5-carbamimidoylsulfanecarbonyl-3,4-dichlorothiophene-2-carbonyl)isothiourea; thiophene-2,5-diylbis(methylene) dicarbamimidothioate; (3,4-diphenylthiophene-2,5-diyl)bis(methylene) dicarbamimidothioate;
(3,4-dimethylthiophene-2,5-diyl)bis(methylene) dicarbamimidothioate;
(3,4-dimethylthieno[2,3-b]thiophene-2,5-diyl)bis(methylene) dicarbamimidothioate;
(4-amino-4H-1 ,2,4-triazole-3,5-diyl)bis(methylene) dicarbamimidothioate; and
(1/-/-1 ,2,4-triazole-3,5-diyl)bis(methylene) dicarbamimidothiodate. Of the above compounds, the following compound, which is described in PCT
Published Application No. WO 2008/151288, is referred to herein as "Compound A":
Figure imgf000034_0001
and is named herein as (2,4,6-trimethyl-1 ,3-phenylene)bis(methylene) dicarbamimidothioate dihydrochloride. This compound is useful as a DMT1 inhibitor in the practice of the invention.
The DMT1 inhibitors disclosed herein can be prepared by the methods disclosed in PCT Published Application No. WO 2008/109840; PCT Published
Application No. WO 2008/115999; PCT Published Application No. WO 2008/118790; PCT Published Application No. WO 2008/121861 ; and PCT Published Application No. WO 2008/151288, the disclosures of which are incorporated in full herein in their entireties. Typically, a successful active ingredient of the present invention, including the
DMT1 inhibitors, will meet some or all of the following criteria. Oral availability should be at less than 5%. Animal model efficacy is less than about 0.1 μg/Kg to about 100 mg/Kg body weight and the target human dose is between 0.1 μg/Kg to about 100 mg/Kg body weight, although dosages outside of this range may be acceptable ("mg/Kg" means milligrams of compound per kilogram of body mass of the subject to whom it is being administered). The potency (as expressed by IC50 value) should be less than 10 μM, preferably below 1 μM and most preferably below 50 nM. The IC50 ("Inhibitory Concentration - 50%") is a measure of the amount of compound required to achieve 50% inhibition of DMT1 , over a specific time period, in an assay of the invention.
Iron Chelators of the Invention
Iron chelators are small molecule ligands that bind iron as Fe3+ with high affinity. The iron bound to the iron chelators is then excreted out of the body typically in the feces, urine or both. Iron chelators are able to induce a net iron excretion in a dose dependent way. Typically, clinically relevant iron excretion is between 0.1 mg/Kg to 0.5 mg/Kg per day.
Some iron chelators have exhibited side effects such as acute renal failure or serum creatinine increases above acceptable and normal levels. The serum creatinine level increases in a dose dependent way. If these side effects are observed, it is recommended that patients receive a dose reduction, interruption or discontinuation of treatment with the iron chelator. For example, for Exjade® (Deferasirox, [4-[(3Z,5E)- 3,5-bis(6-oxo-1-cyclohexa-2,4-dienylidene)-1 ,2,4-triazolidin-1-yl]benzoic acid, Novartis) treated adult patients their daily dose of Exjade® should be reduced by 10 mg/Kg if a >33% rise in serum creatinine should occur at ≥ 2 consecutive post baseline visits. Such action in the chelation treatment regime would reduce the daily net iron excretion in a patient and may leave the patient vulnerable to iron overload disorders or associated diseases. Other side effects associated with most chelation therapy include for example cytopenias, hepatic failures, liver enzyme abnormailites, and skin rashes. Under these circumstances, chelation treatment is also recommended to be dose modified by reducing the daily dose, interrupted or discontinued. Thus, in one aspect the present invention contemplates reducing the incidence of these side effects of chelation therapy by using lower dosages of an iron chelator in the combination therapy. In another aspect, the invention provides a combination therapy for treating iron overload disorders without utilizing an iron chelator in order to avoid the side effects.
Examples of pharmaceutically acceptable iron chelators that may be utilized in the combination therapies of the invention are selected from the group consisting of Deferasirox, Deferoxamine (Desferal®, DFO, Desferoxamine, Λ/-[5-{3-[(5- aminopentyl)hydroxycarbamoyl]propionamido}pentyl]-3-{[5-/V- hydroxyacetamido)pentyl]carbamoyl}propionohydroxamic acid or pharmaceutically acceptable salts thereof, Novartis), Deferiprone (Ferriprox™, (3-hydroxy-1 ,2-dimethyl- 4(1 H)-pyridone), Apotex), Deferitrin (GT-56-252, Genzyme), Dexrazoxane ((+)-( S)-4,4'- propylenedi-2,6-piperazinedione), Desferrithiocin (DFT), Desferri-exochelin (D-Exo), C94, tachpyridine, aroylhydrazones, thiosemicarbazones, pyridoxal isonicotinyl hydrazones, /V,Λ/'-bis(2-hydroxybenzoyl)-ethylenediamine-Λ/,Λ/'-diacetic acid (HBED), and 1-substituted-2-alkyl-3-hydroxy-4-pyridones, including 1-(2'-carboxyethyl)-2- methyl-3-hydroxy-4-pyridone, and other analogs or moieties known by one skilled in the art to chelate iron. Still other pharmaceutically acceptable iron chelators contemplated by the present invention include those described in the following U.S. Patent Application Nos. 10/300,071 ; 10/491 ,310; 10/534,357; 10/544,570; 10/580,011 ; 10/803,724; 11/011 ,110; and 11/367,042; and U.S. Patent Nos. 7,144,904; 7,126,004; 7,074,815; 6,932,960; 6,855,711 ; 6,723,742; 6,623,721 ; 6,589,966; 6,583,182; 6,559,315; 6,541 ,490; 6,525,080; 6,521 ,652; 6,469,162; 6,465,504; 6,455,578; 6,448,273; 6,335,353; 6,083,966; 5,874,573; 5,840,739; 5,834,492; 5,721 ,209; 5,480,894; 5,430,058; 5,322,961 , 4,840,958; 4,684,482; and 4,528,196.
Proton Pump Inhibitors of the Invention
The proton pump inhibitors (PPIs) utilized in the combination therapies of the invention are compounds having H+, K+-ATPase inhibiting activity. PPIs suppress gastric acid secretion by specific inhibition of the H+, K+-ATPase in the gastric parietal cell. This inhibition of gastric acid is thought to lead to a higher pH in the stomach and gut which could lead to a decrease in the solubilization of non-heme iron and subsequently to decrease its absorption into the body. PPIs may, if desired, be in the form of free base, free acid, salt, ester, solvates (in particular hydrates), anhydrate, amide, enantiomer, isomer, tautomer, prodrug, polymorph, or any other pharmacologically suitable derivative, or the like, that is therapeutically active. Examples of PPIs of the invention include, but are not limited to, Omeprazole (Prilosec, AstraZeneca), Esomeprazole (Nexium, AstraZeneca), Dontoprazole, Perprazole, Habeprazole, Ransoprazole, Pariprazole, Lansoprazole (Prevacid, TAP Pharmaceuticals), Pantoprazole (Protonix, Wyeth-Ayerst), Leminoprazole, Nepaprazole, Rabeprazole (Aciphex, Janssen Pharmaceutica), Leminoprazole, Nepaprazole and Tenatoprazole (or Benatoprazole) (Mitsubishi Pharma). Other PPIs, include, but are not limited to: Soraprazan (Atlana); llaprazole (II-
Yang Pharm); AZD-0865 (AstraZeneca); YH-1885 (PCT Published Patent Application No. WO 96/05177); SB-641257 (2-pyrimidinamine, 4-(3,4-dihydro-1-methyl-2(1H)- isoquinolinyl)-Λ/-(4-fluorophenyl)-5,6-dimethyl-monohydrochloride) (YuHan); BY-112 (Altana); SPI-447 (lmidazol(1 ,2-a)thieno(3,2-c)pyridine-3-amine,5-methyl-2-(2-methyl- 3-thieny-l) (Shinnippon); 3-hydroxymethyl-2methyl-9-phenyl-7H-8,9-dihydro-pyrano(2,- 3-c)-imidazo(1 ,2-a)pyridine (PCT Published Patent Application No. WO 95/27714, AstraZeneca); Pharma projects No. 4950 (3-hydroxymethyl-2-methyl-9-phenyl-7H-8,9- dihydro-pyrano(2,3-c)-imidazol(1 ,2-a)pyridine) (AstraZeneca, ceased) (PCT Published Patent Application No. WO 95/27714); Pharmaprojects No. 4891 (EP 700899) (Aventis); Pharmaprojects No. 4697 (PCT Published Patent Application No. WO 95/32959) (AstraZeneca); H-335/25 (AstraZeneca); T-330 (Saitama 335) (Pharmacological Research Lab); Pharmaprojects No. 3177 (Roche); BY-574 (Altana); Pharmaprojects No. 2870 (Pfizer); AU-1421 (EP 264883) (Merck); AU- 2064 (Merck); AY-28200 (Wyeth); Pharmaprojects No. 2126 (Aventis); WY-26769 (Wyeth); pumaprazole (PCT Published Patent Application No. WO 96/05199) (Altana); YH- 1238 (YuHan); Pharmaprojects No. 5648 (PCT Published Patent Application No. WO 97/32854) (Dainippon); BY-686 (Altana); YM-020 (Yamanouchi); GYKI-34655 (Ivax); FPL-65372 (Aventis); Pharmaprojects No. 3264 (EP 509974) (AstraZeneca); nepaprazole (To aEiyo); HN- 11203 (Nycomed Pharma); OPC-22575; pumilacidin A (BMS); saviprazole (EP 234485) (Aventis); SKand F-95601 (GSK, discontinued); Pharmaprojects No. 2522 (EP 204215) (Pfizer); S-3337 (Aventis); RS-13232A
(Roche); AU-1363 (Merck); SKand F-96067 (EP 259174) (Altana); SUN 8176 (Daiichi
Phama); Ro-18-5362 (Roche); ufiprazole (EP 74341 ) (AstraZeneca); and Bay-p-1455
(Bayer). Still other proton pump inhibitors contemplated by the present invention include those described in the following U.S. Patent Nos.: 4,628,098; 4,689,333; 4,786,505;
4,853,230; 4,965,269; 5,021 ,433; 5,026,560; 5,045,321 ; 5,093,132; 5,430,042;
5,433,959; 5,576,025; 5,639,478; 5,703,110; 5,705,517; 5,708,017; 5,731 ,006;
5,824,339; 5,855,914; 5,879,708; 5,948,773; 6,017,560; 6,123,962; 6,187,340; 6,296,875; 6,319,904; 6,328,994; 4,255,431 ; 4,508,905; 4,636,499; 4,738,974;
5,690,960; 5,714,504; 5,753,265; 5,817,338; 6,093,734; 6,013,281 ; 6,136,344;
6,183,776; 6,328,994; 6,479,075; 6,559,167 and 6,617,338.
PPIs as well as their salts, hydrates, esters, amides, enantiomers, isomers, tautomers, polymorphs, prodrugs, and derivatives may be prepared using standard procedures known to those skilled in the art of synthetic organic chemistry. See, e.g.,
March, Advanced Organic Chemistry: Reactions, Mechanisms and Structure, 4th Ed.
(New York: Wiley-lnterscience, 1992); Leonard et al., Advanced Practical Organic
Chemistry (1992); Howarth et a/., Core Organic Chemistry (1998); and Weisermel et ai, Industrial Organic Chemistry (2002). Calcium Channel Blockers of the Invention
Calcium channel blockers utilized in the combination therapies of the invention are compounds having the ability to interact with and block calcium transport through L-type calcium channels located on skeletal and smooth muscles. L-type calcium channel blockers also exert a direct effect on DMT1 , prolonging the activity of DMT1 , thereby stimulating renal iron excretion and decreasing serum iron levels.
Examples of calcium channel blockers that, if present, may be utilized in the combination therapies of the invention are selected from the group consisting of dihydropyridine compounds such as nifedipine, nicardipine, niguldipine and nimopidine. Still other calcium channel blockers contemplated by the present invention include those described in the following U.S. Patent Nos.: 7,485,653; 7,235,570; 7,098,211 ; 6,852,742; 5,143,915; 5,068,337; 5,021 ,436; 4,849,436; 4,841 ,054; and 4,833,150.
The foregoing exemplary list of active ingredients are meant to be illustrative and not exhaustive as a person of ordinary skill in the art would recognize that there are many other suitable DMT1 inhibitors, PPIs, pharmaceutically acceptable iron chelators and calcium channel blockers which could be utilized in the combination therapies of the invention.
EMBODIMENTS OF THE INVENTION Of the various aspects of the invention as set forth above in the Summary of the Invention, one embodiment of the invention is where the pharmaceutically acceptable iron chelator is selected from Deferasirox, Deferoxamine, DFO, Desferoxamine, Deferiprone, Deferitrin, Dexrazoxane, Desferrithiocin, Desferri- exochelin, C94, tachpyridine, aroylhydrazones, thiosemicarbazones, pyridoxal isonicotinyl hydrazones, Λ/,Λ/'-bis(2-hydroxybenzoyl)-ethylenediamine-Λ/,Λ/'-diacetic acid, or 1-substituted-2-alkyl-3-hydroxy-4-pyridones (including 1-(2'-carboxyethyl)-2- methyl-3-hydroxy-4-pyridone).
Another embodiment of the invention is where the pharmaceutically acceptable iron chelator is selected from Deferasirox, Deferiprone, Deferitrin or Deferoxamine. Another embodiment of the invention of the invention is where the proton pump inhibitor is selected from Omeprazole, Esomeprazole, Dontoprazole, Perprazole, Habeprazole, Ransoprazole, Pariprazole, Lansoprazole, Pantoprazole, Leminoprazole, Nepaprazole, Rabeprazole, Leminoprazole, Nepaprazole, Tenatoprazole, Soraprazan, llaprazole, AZD-0865, YH-1885, ,4-(3,4-dihydro-1-methyl-2(1 W)-isoquinolinyl)-Λ/-(4- fluorophenyl)-5,6-dimethyl-2-pyrimidinamine monohydrochloride, BY-112, imidazol(1 ,2- a)thieno(3,2-c)pyridine-3-amine,5-methyl-2-(2-methyl-3-thienyl), 3-hydroxymethyl- 2methyl-9-phenyl-7H-8,9-dihydro-pyrano(2,-3-c)-imidazo(1 ,2-a)pyridine, ,3- hydroxymethyl-2-methyl-9-phenyl-7/-/-8,9-dihydro-pyrano(2,3-c)-imidazol(1 ,2- a)pyridine, Pharmaprojects No. 4891 , Pharmaprojects No. 4697, H-335/25, T-330, Pharmaprojects No. 3177, BY-574, Pharmaprojects No. 2870, AIM 421 , AU- 2064, AY-28200, Pharmaprojects No. 2126, WY-26769, pumaprazole, YH-1238, Pharmaprojects No. 5648, BY-686, YM-020, GYKI-34655, FPL-65372, Pharmaprojects No. 3264, nepaprazole, HN- 11203, OPC-22575, pumilacidin A, saviprazole, SKand F- 95601 , Pharmaprojects No. 2522, S-3337, RS-13232A, AU-1363, SKand F-96067, SUN 8176, Ro-18-5362, ufiprazole, or Bay-p-1455.
Another embodiment of the invention is where the proton pump inhibitor is selected from the group consisting of Omeprazole, Esomeprazole, Lansoprazole, Pantoprazole, Rabeprazole and Tenatoprazole.
Another embodiment of the invention is where the proton pump inhibitor is selected from the group consisting of Omeprazole and Lansoprazole.
Another embodiment of the invention is where the calcium channel blocker is selected from the group consisting of: nifedipine, nicardipine, niguldipine and nimopidine. Another embodiment of the invention is where the calcium channel blocker is nifedipine.
Another embodiment of the invention is where the calcium channel blocker is a
L-type calcium channel blocker.
Another embodiment of the invention is where the DMT1 inhibitor is selected from:
(2,4,6-trimethyl-1 ,3-phenylene)bis(methylene) dicarbamimidothioate; dibenzo[ιb,c(|thiophene-4,6-diylbis(methylene) dicarbamimidothioate;
(2-fluorodibenzo[b,c/]thiophene-4,6-diyl)bis(methylene) dicarbamimidothioate;
(3,7-dibromodibenzo[ib;c/]thiophene-4,6-diyl)bis(methylene) dicarbamimidothioate; (2-chloro-8-fluorodibenzo[jfc),c/]thiophene-4,6-diyl)bis(methylene) dicarbamimidothioate;
(3-bromodibenzo[jb,cf]thiophene-4,6-diyl)bis(methylene) dicarbamimidothioate; dibenzo[/b,d]thiophene-1 ,9-diylbis(methylene) dicarbamimidothioate;
(9-oxo-9H-xanthene-4,5-diyl)bis(methylene) dicarbamimidothioate; dibenzo[b,c/]furan-4,6-diylbis(methylene) dicarbamimidothioate; (3, 7-dimethyldibenzo[tι,cf]furan-4,6-diyl)bis(methylene) dicarbamimidothioate;
(3,7-dichloroldibenzo[b,o(]furan-4,6-diyl)bis(methyiene) dicarbamimidothioate;
(3, 7-dibromodibenzo[/b,c/]furan-4,6-diyl)bis(methylene) dicarbamimidothioate;
(2-fluorodibenzo[b,c/]furan-4,6-diyl)bis(methylene) dicarbamimidothioate;
(2,8-dibromodibenzo[ib;c/]furan-4,6-diyl)bis(methylene) dicarbamimidothioate; biphenylene-1 ,8-diylbis(methylene) dicarbamimidothioate;
(3,6-difluorobiphenylene-1 ,8-diyl)bis(methylene) dicarbamimidothioate; biphenylene-1 ,4,5,8-tetrayltetrakis(methylene) tetracarbamimidothioate;
2-(8-carbamimidoylsulfanylmethyl-9-όxo-9/-/-fluoren-1-ylmethyl)-isothiourea;
(9-oxo-9/-/-fluorene-4,5-diyl)bis(methylene) dicarbamimidothioate; 2-(2,7-di-tert-butyl-5-carbamimidoylsulfanylmethyl-9,9-dimethyl-9/-/-xanthen-4- ylmethyl)-isothiourea; phenoxathiine-4,6-diylbis(methylene) dicarbamimidothioate;
(5,7-dihydrodibenzo[c,e]thiepine-1 ,1 i-diyl)bis(methylene) dicarbamimidothioate;
2-(2'-carbamimidoylsulfanylmethyl-biphenyl-2-ylmethyl)-isothiourea; (6,6'-dimethylbiphenyl-2,2'-diyl)bis(methylene) dicarbamimidothioate dihydrobromide; biphenyl-2,2',6,6'-tetrayltetrakis(methylene) tetracarbamimidothioate; dimethyl 6,6'-bis(carbamimidoylthiomethyl)biphenyl-2,2'-dicarboxylate;
2-[2-(2-carbamimidoylsulfanylmethyl-phenoxy)-benzyl]-isothiourea;
2-(1-{2-[2-(1-carbamimidoylsulfanyl-ethyl)-phenoxy]-phenyl}-ethyl)-isothiourea; 2-[2-({[amino(imino)methyl]thio}methyl)-4-fluorophenoxy]-5-nitrobenzyl imidothiocarbamate;
2-[2-({[amino(imino)methyl]thio}methyl)-5-chlorophenoxy]-4-nitrobenzyl imidothiocarbamate;
2-[2-({[amino(imino)methyl]thio}methyl)-5-chlorophenoxy]-5-fluorobenzyl imidothiocarbamate;
2-[2-(2-carbamimidoylsulfanylmethyl-3-chlorophenoxy)-5-fluorobenzyl]isothiourea;
2-[2-(2-carbamimidoylsulfanylmethyl-4-chlorophenoxy)-5-fluorobenzyl]isothiourea;
2-[2-({[amino(imino)methyl]thio}methyl)-4-chlorophenoxy]benzyl imidothiocarbamate;
2-[2-({[amino(imino)methyl]thio}methyl)-3-chlorophenoxy]benzyl imidothiocarbamate; 2-[2-({[amino(imino)methyl]thio}methyl)-5-chlorophenoxy]benzyl imidothiocarbamate;
2-[2-({[amino(imino)methyl]thio}methyl)-5-chlorophenoxy]-5-nitrobenzyl imidothiocarbamate;
2-[2-({[amino(imino)methyl]thio}methyl)-4-chlorophenoxy]-5-nitrobenzyl imidothiocarbamate; 2-[2-(2-carbamimidoylsulfanylmethyl-5-fluorophenoxy)-5-fluorobenzyl]isothiourea;
2-[2-(2-carbamimidoylsulfanylmethyl-4-fluorophenoxy)-5-fluorobenzyl]isothiourea;
2-[2-({[amino(imino)methyl]thio}methyl)-4-fluorophenoxy]benzyl imidothiocarbamate;
2-[2-(2-carbamimidoylsulfanylmethyl-phenylsulfanyl)-benzyl]-isothiourea;
2-[2-(2-carbamimidoylsulfanylmethyl-4-fluoro-phenylsulfanyl)-5-fluoro-benzyl]- isothiourea;
2-[2-(2-carbamimidoylsulfanylmethyl-4-fluoro-phenylsulfanyl)-benzyl]-isothiourea;
2-[2-(2-carbamimidoylsulfanylmethyl-5-methyl-phenylsulfanyl)-benzyl]-isothiourea;
2-[2-(2-carbamimidoylsulfanylmethyl-4-methoxy-phenylsulfanyl)-benzyl]-isothiourea;
2-[2-(2-carbamimidoylsulfanylmethyl-5-methyl-phenylsulfanyl)-5-fluoro-benzyl]- isothiourea;
2-[2-(2-carbamimidoylsulfanylmethyl-4-methoxy-phenylsulfanyl)-5-fluoro-benzyl]- isothiourea;
2-[2-(2-carbamimidoylsulfanylmethyl-5-chloro-phenylsulfanyl)-5-fluoro-benzyl]- isothiourea; 2-[2-(2-carbamimidoylsulfanylmethyl-6-methylphenylsulfanyl)-benzyl]isothiourea; 2-[2-(2-carbamimidoylsulfanylmethyl-4,5-difluorophenylsulfanyl)-benzyl]isothiourea; 2-[2-(2-carbamimidoylsulfanylmethyl-4-methyl-phenylsulfanyl)-benzyl]-isothiourea; 2-[2-(2-carbamimidoylsulfanylmethyl-5-chloro-phenylsulfanyl)-benzyl]-isothiourea; 2-[2-(2-carbamimidoylsulfanylmethyl-5-fluoro-phenylsulfanyl)-benzyl]-isothiourea; 2-[2-(2-carbamimidoylsulfanylmethyl-5-fluoro-phenylsulfanyl)-5-fluorobenzyl]- isothiourea; 2-[2-(2-carbamimidoylsulfanylmethyl-4,5-difluorophenylsulfanyl)-5- fluorobenzyφsothiourea;
2-{[2-({[amino(imino)methyl]thio}methyl)-4-fluorophenyl]thio}-3-nitrobenzyl imidothiocarbamate;
2-{[2-({[amino(imino)methyl]thio}methyl)-4-fluorophenyl]thio}-5-nitrobenzyl imidothiocarbamate; 2-[2-(2-carbamimidoylsulfanylmethyl-4-fluorophenylsulfanyl)-5-
(trifluoromethyl)benzyl]isothiourea; 2-[2-(2-carbamimidoylsulfanylmethylphenylsulfanyl)-5-aminobenzyl]-isothiourea; 2-[2-(2-carbamimidoylsulfanylmethyl-4-chlorophenylsulfanyl)-5- fluorobenzyφsothiourea;
2-[2-(2-carbamimidoylsulfanylmethylphenylsulfanyl)-5-ethylaminobenzyl]isothiourea; 2-[2-(2-carbamimidoylsulfanylmethyl-4-chlorophenylsulfanyl)-5- chlorobenzyljisothiourea;
2-[2-(2-carbamimidoylsulfanylethyl-4-fluorophenylsulfanyl)-5-fluorobenzyl]isothiourea; 2-[2-(2-carbamimidoylsulfanylethyl-4-chlorophenylsulfanyl)-5-fluorobenzyl]isothiourea; 2-[2-(2-carbamimidoylsulfanylethylphenylsulfanyl)benzyl]isothiourea; 2-[2-(2-carbamimidoylsulfanylmethylphenylsulfanyl)-5-chlorobenzyl]-isothiourea; 2-[2-(2-carbamimidoyIsulfanylmethylphenylsulfanyl)-5-
(trifluoromethyl)benzyl]isothiourea;
2-[2-(2-methylcarbamimidoylsulfanylmethylphenylsulfanyl)benzyl]-methylisothiourea; 2-[2-(2-carbamimidoylsulfanylmethyl-4-fluorophenylsulfanyl)-5-
(methylsulfonyl)benzyl]isothiourea; 2-({2-({[amino(imino)methyl]thio}methyl)-4-[(dimethylamino)sulfonyl]phenyl}thio)-5- fluorobenzyl imidothiocarbamate;
2-[2-(2-carbamimidoylsulfanylmethylphenylsulfanyl)benzyl]-isothiourea; 2-{[2-({[amino(imino)methyl]thio}methyl)-4-fluorophenyl]thio}-4-(methylsulfonyl)benzyl imidothiocarbamate; 2-{[2-({[amino(imino)methyl]thio}methyl)-4-chlorophenyl]thio}-5-cyanobenzyl imidothiocarbamate;
2-[2-(2-carbamimidoylsuifanylmethylphenylsulfonyl)benzyl]-isothiourea;
2-[2-(2-carbamimidoylsulfanylmethyl-4-fluorophenylsulfonyl)-5-fluorobenzyl]isothiourea;
2-(6-((aminoamidino)thiomethyl)phenyl)thio-1-((aminoamidino)thiomethyl)benzene; 2-(6-(2-amidinoethyl)phenyl)thio-1 -(2-amidinoethyl)benzene;
2-(6-(amidinomethyl)phenyl)thio-1-(amidinomethyl)benzene;
2-(6-(3-amidinopropyl)phenyl)thio-1-(3-amidinopropyl)benzene;
2-(6-((cyanoamidino)methyl)phenyl)thio-1-((cyanoamidino)methyl)benzene;
2-(6-(2-(cyanoamidino)ethyl)phenyl)thio-1-(2-(cyanoamidino)ethyl)benzene; 2-(6-(3-(cyanoamidino)propyl)phenyl)thio-1 -(3-(cyanoamidino)propyl)benzene;
1-(2-(2-(guanidinomethyl)phenylthio)benzyl)guanidine;
2,2'-thiobis(Λ/-(diaminomethylene)benzamide);
2-(6-(amidinothiomethyl)phenyl)carbonyl-1-(amidinothiomethyl)benzene;
2-(6-(amidinothiomethyl)phenyl)carbonyl-1-(amidinothiomethyl)-5-fluorobenzene; 2-(6-(amidinothiomethyl)phenyl)carbonyl-1 -(amidinothiomethyl)-4-fluorobenzene;
2-(6-(amidinothiomethyl)phenyl)carbonyl-1-(amidinothiomethyl)-4-chlorobenzene;
2-(6-(amidinothiomethyl)phenyl)carbonyl-1-(amidinothiomethy!)-5-chlorobenzene;
4,4-diisothiourea benzophenone;
2,2-(methylazanediyl)bis(2,1-phenyiene)bis(methylene)dicarbamimidothioate; 2-[2-(1-carbamimidoylsulfanylmethyi-naphthalen-2-ylsulfanyl)-benzyl]-isothiourea;
2-[2-(1-carbamimidoylsulfanylmethylnaphthalen-2-ylsulfanyl)-5-fluorobenzyl]- isothiourea;
(2-{[2-({[amino(imino)methyl]thio}methyl)-4-fluorophenyl]thio}-3-thienyl)methyl imidothiocarbamate; (4-{[2-({[amino(imino)methyl]thio}methyl)-4-fIuorophenyl]thio}-3-thienyl)methyl imidothiocarbamate;
2-pyridin-2-yl-4,5-dihydro-2/-/-benzo[e]indazoi-1-ol;
7-bromo-2-pyridin-2-yl-4,5-dihydro-2/-/-benzo[e]indazol-1-ol;
2-pyridin-2-yl-2,4,5,6-tetrahydro-2,3-diaza-benzo[e]azulen-1-ol; 1-hydroxy-2-pyridin-2-yl-4,5-dihydro-2/-/-benzo[e]indazole-7-carbonitrile;
7-phenyl-2-pyridin-2-yl-4,5-dihydro-2/-/-benzo[e]indazol-1-ol;
7-benzo[1 ,3]dioxol-5-yl-2-pyridin-2-yl-4,5-dihydro-2/-/-benzo[e]indazol-1-ol;
4-(1-hydroxy-2-pyridin-2-yl-4,5-dihydro-2/-/-benzo[e]indazol-7-yl)-benzonitrile;
2-pyridin-2-yl-7-p-tolyl-4,5-dihydro-2H-benzo[e]indazol-1-ol; 7-(4-methoxyphenyl)-2-pyridin-2-yl-4,5-dihydro-2/-/-benzo[e]indazol-1-ol; 2-(1-hydroxy-4,5-dihydro-2/-/-benzo[e]indazol-2-yl)pyridine 1 -oxide;
7-methoxy-2-pyridin-2-yl-4,5-dihydro-2/-/-benzo[e]indazol-1-ol;
8-methoxy-2-pyridin-2-yl-4,5-dihydro-2/-/-benzo[e]indazol-1-ol;
2-(5-(trifluoromethyl)pyridin-2-yl)-4,5-dihydro-2/-/-benzo[e]indazol-1-ol; 2-(4-(trifluoromethyl)pyridin-2-yl)-4,5-dihydro-2/-/-benzo[e]indazol-1-ol;
2-(5-nitropyridin-2-yl)-4,5-dihydro-2H-benzo[e]indazol-1-ol;
2-(3-(trifluoromethyl)pyridin-2-yl)-4,5-dihydro-2/-/-benzo[e]indazol-1-ol;
2-(5-methylpyridin-2-yl)-4,5-dihydro-2H-benzo[e]indazol-1-ol;
2-(benzo[c/]thiazol-2-yl)-4,5-dihydro-2/-/-benzo[e]indazol-1-ol; 2-(6-methoxybenzo[cf|thiazol-2-yl)-4,5-dihydro-2H-benzo[e]indazol-1 -ol;
2-(6-fluorobenzo[cf]thiazol-2-yl)-4,5-dihydro-2/-/-benzo[e]indazol-1-ol;
2-(benzo[cy]thiazol-2-yl)-7-methoxy-4,5-dihydro-2/-/-benzo[e]indazol-1-ol;
2-(6-methylbenzo[d]thiazol-2-yl)-4,5-dihydro-2/-/-benzo[e]indazol-1-ol;
2-(4-methylbenzo[c/]thiazol-2-yl)-4,5-dihydro-2/-/-benzo[e]indazol-1-ol; 2-(1 -hydroxy^.δ-dihydro^H-benzo^indazol^-yObenzofc/lthiazole-θ-carboxylic acid;
2-(1 /-/-benzo[c/|imidazol-2-yl)-4,5-dihydro-2/-/-benzo[e]indazol-1-ol;
2-(4-tert-butylthiazol-2-yl)-4,5-dihydro-2/-/-benzo[e]indazol-1-ol hydrochloride;
1-(2-bromophenyl)-3-methyl-4-phenyl-1/-/-pyrazol-5-amine;
3-methyl-1-(pyridin-2-yl)-4-(4-(trifluoromethyl)phenyl)-1 /-/-pyrazol-5-ol; 3-methyl-4-phenyl-1 -(pyridin-2-yl)-1 /-/-pyrazol-5-amine;
2-(3,5-dimethyl-4-phenyl-1/-/-pyrazol-1-yl)pyridine;
3-methyl-4-phenyl-1 -(pyridin-2-yl)-1 H-pyrazoI-5-yl trifluoromethanesulfonate;
3-methyl-1-(pyridin-2-yl)-4-(3-(trifluoromethyl)phenyl)-1 /-/-pyrazol-5-ol;
4-(5-hydroxy-3-methyl-1 -(pyridin-2-yl)-1 /-/-pyrazol-4-yl)benzoic acid; 4-(5-hydroxy-3-methyl-1 -(pyridin-2-yi)-1 /-/-pyrazol-4-yl)-Λ/-(6-hydroxyhexyl)benzamide;
3-methyl-4-(4-(methylsulfonyl)phenyl)-1-(pyridin-2-yl)-1/-/-pyrazol-5-ol;
4-(4-(hydroxymethyl)phenyl)-3-methyl-1-(pyridin-2-yl)-1/-/-pyrazol-5-ol;
3-methyl-4-phenyl-1 -(pyrimidin-2-yl)-1 H-pyrazol-5-ol;
6-(5-hydroxy-3-methyl-4-phenylpyrazol-1-yl)-pyridazin-3-ol; 3-methyl-4-phenyl-1 -(thiazol-2-yl)-1 H-pyrazol-5-ol;
1 -(benzo[d]thiazol-2-yl)-3-methyl-4-phenyl-1 H-pyrazol-5-ol;
1 -(1 H-benzo[αφmidazol-2-yl)-3-methyl-4-phenyl-1 /-/-pyrazol-5-ol;
1 -(isoquinolin-1 -yl)-3-methyl-4-phenyl-1 /-/-pyrazol-5-ol;
1-(6-methoxybenzo[c/]thiazol-2-yl)-3-methyl-4-phenyl-1 H-pyrazol-5-ol; 1 -(6-fluorobenzo[d]thiazol-2-yl)-3-methyl-4-phenyl-1 H-pyrazol-5-ol; 3-methyl-1 -(5-nitrothiazol-2-yl)-4-phenyl-1 H-pyrazol-5-ol;
3-methyl-4-(4-nitrophenyl)-1 -(pyridin-2-yl)-1 H-pyrazol-5-ol;
4-(4-aminophenyl)-3-methyl-1-(pyridin-2-yl)-1 H-pyrazol-5-ol;
4-(4-methoxyphenyl)-3-methyl-1-(pyridin-2-yl)-1H-pyrazol-5-oi; 4-(4-(diethylamino)phenyl)-3-methyl-1 -(pyridin-2-yl)-1 H-pyrazol-5-ol;
4-(5-hydroxy-3-methyl-1-(pyridin-2-yl)-1H-pyrazol-4-yl)benzonitrile;
4-(4-fluorophenyl)-3-methyl-1-(pyridin-2-yl)-1 H-pyrazol-5-ol; ethyl 4-(5-hydroxy-3-methyl-1 -(pyridin-2-yl)-1 H-pyrazol-4-yl)benzoate;
3-methyl-4-phenyl-1-(pyridin-2-yl-methyl)-1 H-pyrazol-5-ol; 1 ,3-phenylenebis(methylene) dicarbamimidothioate;
(2-fluoro-1 ,3-phenylene)bis(methylene) dicarbamimidothioate;
1 ,3-phenylene dicarbamimidothioate;
(5-methyl-1 ,3-phenylene)bis(methylene) dicarbamimidothioate;
(2,4,6-trimethylbenzene-1 ,3,5-triyl)tris(methylene) tricarbamimidothioate; 2-{1-[3-(1-carbamimidoylsulfanyl-1-methylethyl)phenyl]-1-methylethyl}isothiourea;
(2-cyano-1 ,3-phenylene)bis(methylene) dicarbamimidothioate;
(4,6-dimethyl-1 ,3-phenylene)bis(methylene) dicarbamimidothioate; diethyl 4,6-bis(carbamimidoylthiomethyl)isophthalate;
(5-bromo-4,6-dimethyl-1 ,3-phenylene)bis(methylene) dicarbamimidothioate; (2,4,5,6-tetramethyl-1 ,3-phenylene)bis(methylene) dicarbamimidothioate;
2-{1-[3-(1-carbamimidoylsulfanylethyl)-2,4,6-trimethylphenyl]ethyl}isothiourea;
(2-hydroxy-5-methyl-1 ,3-phenylene)bis(methylene) dicarbamimidothioate;
1 ,3-di[(methylamidino)thiomethyl]-2,4,6-trimethylbenzene;
(5-hydroxy-2>4,6-trimethyl-1 ,3-phenylene)bis(methylene) dicarbamimidothioate; (2,4,5,6-tetrachloro-1 ,3-phenylene)bis(methylene) dicarbamimidothioate;
(2-methoxy-5-methyl-1 ,3-phenylene)bis(methylene) dicarbamimidothioate;
(2-methyl-1 ,3-phenylene)bis(methylene) dicarbamimidothioate;
(4-methoxy-1 ,3-phenylene)bis(methylene) dicarbamimidothioate;
(5-methoxy-1 ,3-phenylene)bis(methylene) dicarbamimidothioate; (4,6-dibromo-1 ,3-phenylene)bis(methylene) dicarbamimidothioate;
(4,6-diisopropyl-1 ,3-phenylene)bis(methylene) dicarbamimidothioate;
1 ,3-di[(2-cyano-3-methylguanidino)methyl]-2,4,6-trimethylbenzene;
2,2'-(1 ,3-phenylene)diacetimidamide;
A/-(3-guanidinomethyl-2,4,6-trimethylbenzyl)guanidine; pyridine-2,6-diylbis(methylene) dicarbamimidothioate; (2,4>6-trimethylpyridine-3,5-diyl)bis(methylene) dicarbamimidothioate;
(1 ,2-phenylene)bis(methylene) dicarbamimidothioate;
(3,4,5,6-tetramethyl-1 ,2-phenylene)bis(methylene) dicarbamimidothioate; naphthalene-1 ,2-diylbis(methylene) dicarbamimidothioate; naphthalene-1 ,8-diylbis(methylene) dicarbamimidothioate;
2-(5-carbamimidoylsulfanecarbonyl-3,4-dichlorothiophene-2-carbonyl)isothiourea; thiophene-2,5-diylbis(methylene) dicarbamimidothioate;
(3,4-diphenylthiophene-2,5-diyl)bis(methylene) dicarbamimidothioate;
(3,4-dimethylthiophene-2,5-diyl)bis(methylene) dicarbamimidothioate; (3,4-dimethylthieno[2,3-b]thiophene-2,5-diyl)bis(methylene) dicarbamimidothioate;
(4-amino-4H-1 ,2,4-triazole-3,5-diyl)bis(methy!ene) dicarbamimidothioate; or
(1 H- 1 ,2,4-triazole-3,5-diyl)bis(methylene) dicarbamimidothiodate; or pharmaceutically acceptable salts thereof.
Another embodiment of the invention is where the DMT1 inhibitor is (2,4,6- trimethyl-1 ,3-phenylene)bis(methylene) dicarbamimidothioate dihydrochloride.
Another embodiment of the invention is wherein the therapeutically effective amount of the DMT1 inhibitor is a daily dosage amount from about 0.001 mg/Kg body weight to about 100 mg/Kg body weight of the mammal.
Another embodiment of the invention is wherein daily dosage amount of the DMT1 inhibitor is selected from about 10 mg/Kg body weight, about 25 mg/Kg body weight, or about 50 mg/Kg body weight of the mammal.
Another embodiment of the invention is wherein the daily dosage amount of the pharmaceutically acceptable iron chelator is from about 10 mg/Kg body weight to about
100 mg/Kg body weight of the mammal. Another embodiment of the invention is wherein the daily dosage amount of the pharmaceutically acceptable iron chelator is selected from about 10 mg/Kg body weight, about 20 mg/Kg body weight, about 30 mg/Kg body weight, about 50 mg/Kg body weight, about 75 mg/Kg body weight or about 100 mg/Kg body weight of the mammal. Another embodiment of the invention is wherein the therapeutically effective amount of the proton pump inhibitor is a daily dosage amount from about 10 mg/Kg body weight to about 100 mg/Kg body weight of the mammal.
Another embodiment of the invention is wherein the daily dosage amount of the proton pump inhibitor is selected from about 15 mg/Kg body weight, about 40 mg/Kg body weight, or about 75 mg/Kg body weight of the mammal. Another embodiment of the invention is wherein the therapeutically effective amount of the calcium channel blocker is a daily dosage amount from about 0.001 mg/Kg body weight to about 100 mg/Kg body weight of the mammal.
Another embodiment of the invention is wherein the daily dosage amount of the calcium channel blocker is selected from about 0.1 mg/Kg body weight, about 2.0 mg/Kg body weight, or about 20 mg/Kg body weight of the mammal.
Another embodiment of the invention is wherein the therapeutically effective amount of the DMT1 inhibitor is a daily dosage amount from about 0.001 mg/Kg body weight to about 100 mg/Kg body weight of the mammal and the therapeutically effective amount of the proton pump inhibitor is a daily dosage amount from about 10 mg/Kg body weight to about 100 mg/Kg body weight of the mammal.
Another embodiment of the invention is wherein the daily dosage amount of the DMT1 inhibitor is selected from the group consisting of 10 mg/Kg body weight, 25 mg/Kg body weight, and 50 mg/Kg body weight of the mammal, and the daily dosage amount of the proton pump inhibitor is selected from the group consisting of 15 mg/Kg body weight, 40 mg/Kg body weight, and 75 mg/Kg body weight of the mammal.
Another embodiment of the invention is wherein the daily dosage amount of the DMT1 inhibitor is 10 mg/Kg body weight of the mammal and the daily dosage amount of the proton pump inhibitor is 15 mg/Kg body weight of the mammal. Another embodiment of the invention is wherein the daily dosage amount of the
DMT1 inhibitor is 25 mg/Kg body weight of the mammal and the daily dosage amount of the proton pump inhibitor is 40 mg/Kg body weight of the mammal.
Another embodiment is wherein the daily dosage amount of the DMT1 inhibitor is 50 mg/Kg body weight of the mammal and the daily dosage amount of the proton pump inhibitor is 75 mg/Kg body weight of the mammal.
Another embodiment of the invention is wherein all of the active ingredients are concomitantly administered to the mammal.
Another embodiment of the invention is wherein all of the active ingredients are sequentially administered to the mammal. Another embodiment of the invention is wherein at least two of the active ingredients are concomitantly administered to the mammal.
Another embodiment of the invention is wherein at least two of the active ingredients are sequentially administered to the mammal.
Another embodiment is wherein the DMT1 inhibitor and the proton pump inhibitor are concomitantly administered to the mammal. Another embodiment of the invention is wherein the DMT1 inhibitor and the proton pump inhibitor are sequentially administered to the mammal.
Another embodiment of the invention is where the iron overload disorder is a primary iron overload disorder. Another embodiment of the invention is where the primary iron overload disorder is independently selected from hereditary hemochromatosis, juvenile hemochromatosis, ferroportin disease, neonatal hemochromatosis, Bantu siderosis, African iron overload, gracile syndrome, or Friedreich Ataxia.
Another embodiment of the invention is where the primary iron overload is hereditary hemochromatosis.
Another embodiment of the invention is where the iron overload disorder is a secondary iron overload disorder.
Another embodiment of the invention is where the iron overload disorder is transfusional iron overload disorder. Another embodiment of the invention is where the iron overload disorder is associated with a disease and/or condition independently selected from thalassemia (beta and alpha, major, minor and intermedia), hypochromic microcytic anemia, sickle cell anemia, microcytic iron loading anemia, hereditary sideroblastic anemia, congenital dyserythropoeitic anemia, porphyria cutanea tarda, pyruvate kinase deficiency, hereditary atransferrinemia, ceruloplasmin deficiency, myelodysplastic syndromes, pulmonary hemosiderosis, aceruloplasminemia or x-linked sideroblastic anemia.
Another embodiment of the invention is where the iron overload disorder is associated with a disease and/or condition independently selected from neurodegenerative disease (including ALS, prion diseases, Parkinson's, and Alzheimers), cardiovascular disease (including atherosclerosis, ischemic cerebrovascular disease and ischemic stroke), inflammation (including arthritis and disease progression in viral hepatitis), cancer, insulin resistance, non-alcoholic liver disease, alcoholic liver disease, or infectious disease (including HIV, malaria and Yersinia infections).
It is understood that any of the foregoing embodiments of the invention may be combined with any one or more of the foregoing embodiments to arrive at an embodiment of the invention not specifically disclosed herein. Such embodiments are considered to be within the scope of the invention. Specific embodiments of the invention are described in more detail below in the following sections.
UTILITY AND TESTING OF THE COMBINATION THERAPIES OF THE INVENTION
The present invention is directed to the use of certain DMT1 inhibitors in combination with pharmaceutically acceptable iron chelators, calcium channel blockers and/or proton pump inhibitors in treating iron overload disorders in mammals, preferably in human.
The term "iron overload disorder" refers to a condition in a mammal, preferably a human, wherein the level of iron in the body is elevated compared to the normal level of iron for the particular mammal, such as an elevated iron serum level compared to the normal iron serum level for the mammal or an increased level of iron in the liver of the mammal as compared to the normal level of iron in the liver in the mammal. Elevated serum iron levels can be determined by direct measurement of serum iron using a colorimetric assay, or by the standard transferrin saturation assay (which reveals how much iron is bound to the protein that carries iron in the blood), or by the standard serum ferritin assay. For example, transferrin saturation levels of 45% or higher are usually indicative of abnormally high levels of iron in the serum. Elevated iron levels in the liver can be determined measuring the iron content of the liver from tissue obtained by a liver biopsy or by imaging technique such as MRI and/or SQUID. The degree of iron levels in other tissues (e.g., brain, heart) may also be estimated using these and other imaging techniques.
Preferably, the iron overload disorder is a primary iron overload disorder (including, but not limited to, hereditary hemochromatosis, juvenile hemochromatosis, ferroportin disease, neonatal hemochromatosis, Bantu siderosis, African iron overload, gracile syndrome, and Friedreich Ataxia, as well as all of the anemias listed below in which patients may not be transfused but may become iron overloaded due to increased erythroid drive and the resulting increased iron absorption in the gut) and secondary (or transfusional) iron overload disorder which can be caused by repeated transfusions used to treat a number of distinct anemias, including, but not limited to, thalassemia (beta and alpha, major, minor and intermedia), hypochromic microcytic anemias, sickle cell anemia, microcytic iron loading anemias, hereditary sideroblastic anemias, congenital dyserythropoeitic anemias, porphyria cutanea tarda, pyruvate kinase deficiency, hereditary atransferrinemia, ceruloplasmin deficiency, myelodysplastic syndromes, pulmonary hemosiderosis, aceruloplasminemia and x- linked sideroblastic anemia. Divalent metal transporter-1 (DMT1 ), also known as natural resistance- associated macrophage protein-2 (NRAMP2) and divalent cation transporter-1 (DCT1 ), is an expressed transmembrane protein involved in the maintenance of iron levels in the body. DMT1 is particularly important for iron absorption in the duodenum of the small intestine, where it is localized in the cytoplasm and brush border membrane of the villus enterocytes and mediates the influx of dietary non-heme iron from the intestinal lumen into the enterocytes (Gunshin et al., J. CHn. Invest, 2005, 115:1258- 1266). Once dietary iron is absorbed across the intestinal wall, there is no known physiologic mechanism for excreting iron from the body. Thus, excess absorbed iron is largely retained in the body and can accumulate throughout life. Excess accumulation of iron leads to considerable tissue damage and increased subsequent disease risk such as, for example, cirrhosis or hepatocellular carcinoma. Therefore, DMT1 is a primary focal point of controlling intestinal iron absorption for the maintenance of body iron homeostatsis. There is evidence that the upregulation (i.e., increased activity) of DMT1 has a role in iron overload disorders caused by genetic abnormalities, such as hereditary hemochromatosis. Hereditary hemochromatosis is an iron overload disorder due to intestinal iron hyperabsorption. Hereditary hemochromatosis is characterized by a slow accumulation of iron from the diet to toxic levels resulting in tissue injury and multi-organ malfunction. Patients, typically men, develop symptoms of hemochromatosis in their fourth and fifth decade with variable combinations of cirrhosis, hepatoma, arthritis, hypogonadism, diabetes mellitus and cardiomyopathy. The biochemical profile shows elevated transferrin saturation above 45% and a high serum ferritin. The underlying genetic defect in hereditary hemochromatosis is a mutation in the hemochromatosis gene (HFE) on chromosome 6p21. 90% of Northern Europeans with hereditary hemochromatosis are homozygous for a single missense mutation, C282Y in exon 4 of the HFE gene.
DMT1 activity has also been implicated in the etiology and pathophysiology of hypochromic microcytic anemias, thalassemia, microcytic iron loading anemias, hereditary sideroblastic anemias, hereditary hypochromic anemias, congenital dyserythropoietic anemias, pyruvate kinase deficiency, hereditary atransferrinemia, and certain myelodysplastic syndromes, as there is a direct correlation between the degree of iron limited anemia, increased DMT1 expression in the duodenum and, by extension, increased iron absorption via DMT1 (Morgan et al., Blood Cells Molecules and Diseases, 2002, 29:384-399). There is also evidence that DMT1 has a role in iron overload disorders such as acquired iron overload. The risk factors for acquired iron overload might include for example excessive ingestion of red meat, iron supplements or foods that are iron fortified. Acquired iron overload can also occur from the use of iron cookware, drinking unpurified tap water, use of oral contraceptives, blood transfusions and cigarette smoking. DMT1 pattern of expression and function supports it as a candidate target for the treatment of acquired iron overload and other related maladies.
In addition to the small intestine, DMT1 is also highly expressed in the kidney suggesting a role in renal iron handling and possibly reabsorption of filtered iron (Ferguson et al., Am. J. Physiol. Renal. Physiol., 2001 , 280: F803-F814) and is also involved in the delivery of iron to peripheral tissues by transferrin (Fleming et al., Proc. Natl. Acad. Sci., 1998, 85:1148-1153). DMT1 inhibitors, when dosed in a fashion that increases their systemic exposure, may be useful in an acute unloading of iron via the urine, by inhibiting DMT1 expressed in the kidney Other reports in the literature suggest that DMT1 may not play a crucial role in regulating urinary iron excretion. For example, studies performed in the Belgrade rat, which carries a missense neomorphic mutation in DMT1 , G185R, demonstrated that the resultant loss of iron transporting activity in DMT1 had no effect on urinary iron excretion (Ferguson et al., Kidney International (2003) 64:1755-1764). However, since G185R affects DMT1 function by greatly reducing iron transport but by increasing calcium transport activity, these results must be interpreted with caution (Xu et al., PIoS Biology (2004) 2:0378-0386). Indeed, Ferguson et al. noted increased urinary calcium in their experiments in the Belgrade rat.
In addition, dietary studies suggest that the creation of secondary iron overload in rats, using a high iron diet, results in decreased DMT1 expression and increased urinary iron output (Wareing et al., AJP - Renal (2003) 285:1050-1059), in contrast to the findings of Ludwiczek et al., who suggest that an increase in DMT1 function via L- type calcium channel blockers acts to increase urinary iron output. Thus, it is unclear from these reports whether a DMT1 inhibitor or DMT1 agonist would be of use in increasing urinary iron. This discrepancy in results may be explained by the fact that there are reports of multiple cell types expressing DMT1 in the kidney; it is possible that dietary iron overload specifically decreases DMT1 expression in regions of the kidney that are involved in iron reabsorption and that nifedipine agonizes DMT1 function in a fashion that results in a net increase in urinary iron excretion. Alternatively, it could be that L-type Calcium channel blockers exert their effect on urinary iron excretion via some mechanism other than DMT1 , for example by modulating iron flux via L-type voltage gated ion channels in the kidney, as has been observed for iron permeation in cardiomyoctes (Oudit et al., Nat. Med., 2003, 9:1187- 1193; Zhao et al., Kidney International (2002) 61 : 1393-1406.) Finally, alteration of calcium flux in the kidney via L-type voltage gated iron channels could reduce iron reabsorption via DMT1 or other mechanisms, as calcium has been shown to affect iron uptake in other cell types (Barton et al., Gastroenterology (1983) 84:90-101 ; Ci et al., Cell Calcium (2003) 33:257-266). This work would suggest that inhibition of L-type calcium channels in the kidney could reduce iron reabsorption, leading to increased urinary iron output.
DMT1 may also play a role in regulating iron flux to the brain. As there is some indication that iron overload in the brain may play a role in brain pathology, such as Alzheimer's, DMT1 inhibitors may act to reduce the amount of iron absorbed by the brain, when dosed in a fashion that increases their systemic exposure and allows them to play a role at the blood brain barrier or within the brain (Lehmann et al., 2006, J. Med. Genet, 2006, 43(10):e52; Schenck et al., Top. Magn Reson. Imaging., 2006,17(1 ):41-50).
Studies show that mutant mice that are defective in DMT1 activity (mk/mk) develop hyprochromic microcytic anemia, a severe form of iron deficiency anemia, due to a defect in intestinal iron absorption. In contrast, the hfe';' knockout mouse model of hereditary hemochromatosis is characterized by an enhanced intestinal iron uptake and total body iron overload. The hfe'imk/mk double mutant mouse, which carries mutations in both the HFE and DMT1 genes, fails to load iron, indicating that hemochromatosis {hfe';') can be prevented by blocking the flux of iron through the DMT1 protein (Levy et al., J. Clin. Invest, 2000, 105:1209-16). In addition, studies of human patients with hereditary hemochromatosis show that DMT1 is inappropriately upregulated at the intestinal brush border. This aberrant excessive expression of DMT1 in hereditary hemochromatosis is fundamental to the primary pathophysiology of this condition (Zoller et al., Gastroenterology, 2001 , 120:1412-1419). These findings have made DMT1 a therapeutic target for the treatment of iron overload disorders in general, and, in particular, for the treatment of hereditary hemochromatosis. In further support of DMT1 as a therapeutic target in the treatment of iron overload, it has been shown in clinical studies that the majority of the excess iron burden is absorbed in the form of ferrous (non-heme) iron, as opposed to heme-iron (Lynch et al., Blood, 1989, 74:2187-2193). While not wishing to be bound to any particular mechanism of action, DMT1 inhibitors of the invention are useful in treating iron disorders by directly interacting with a region of the DMT1 protein that modulates or controls iron flux. A direct interaction is supported by electrophysiology studies and the fact that the DMT1 inhibitors of the invention are not potent inhibitors of cation flux in the closely related transporter Natural Resistance-Associated Macrophage Protein-1 (NRAMP1 ). The DMT1 inhibitors of the invention modulate the activity of DMT1 downwards, thereby inhibiting the ability of DMT1 to uptake non-heme iron across the cellular membrane. The combination therapies of the invention are therefore useful in treating iron overload disorders which can be ameliorated by the inhibition of DMT1 activity.
The combination therapies of the invention are also useful in treating or preventing symptoms, diseases and/or conditions in a mammal associated with hereditary hemochromatosis due to accumulation of iron in body tissues such as arthritis, liver disease, heart disease, impotence, early menopause, abnormal skin pigmentation, thyroid deficiency, damage to pancreas, diabetes, and damage to adrenal gland (Sheth et al., Annu. Rev. Med., 2000, 51 :443-464).
The combination therapies of the invention are also useful in treating other forms of hemochromatosis including, but are not limited to, juvenile hemochromatosis and neonatal hemochromatosis. Juvenile hemochromatosis has a much earlier onset and exhibits more severe symptoms such as endocrine dysfunction, joint disease, and cardiac abnormalities due to excessive iron deposition from an early age. Neonatal hemochromatosis is a rare fetal gestational condition that results in iron accumulation in the liver of the fetus.
The combination therapies of the invention are also useful in treating or preventing transfusional iron overload. Chronic blood transfusion is the established therapy for thalassaemia major, bone marrow failure and complications of sickle cell anaemia and other related disorders. With hypertransfusion, the systemic iron load accumulates. Because there is no natural way for the body to eliminate the iron, the excess iron in the transfused blood builds up to cause iron overload and becomes toxic to tissues and organs, particularly the liver, heart, and pancreas. Transfusional iron overload typically results in the patient's premature death from organ failure. The transfusional iron overload is unfortunately augmented by increased iron absorption, which is the natural attempt of the body to increase iron levels in order to promote erythropoiesis, which is itself compromised by the disease states above. Decreased absorption of iron by the inhibition of DMT1 activity may reduce the iron overload related to the transfusional iron overload and supports the use of DMT1 inhibitors in the combination therapies of the invention.
The combination therapies of the invention may be useful in reducing the daily dosage of chelation therapy and preventing the chelation therapy associated side effects in patients including, but not limited to, those with iron overload and renal impairment, and without a corresponding reduction in daily net excreted iron.
The combination therapies of the invention may also be useful in delaying when younger patients are required to begin chelation therapy and thereby protecting them from chelation associated toxicities by blocking the uptake of iron from the gut. The combination therapies of the invention may be useful in reducing the daily dosage of PPIs and preventing the PPI associated side effects in patients, particularly those on long term use, including, but not limited to, those with iron overload and renal impairment, and without a corresponding reduction in daily net excreted iron.
It is expected that combination therapies of the present invention will exert greater than additive effects or synergistic effects as the mechanisms of action employed for the active ingredients are different and each will probably act independently of one another. Each active ingredient will act upon a different pool of iron. For example, PPIs may limit gut uptake by reducing non-heme iron solubility in the gut, whereas DMT1 inhibitors will block the uptake of iron in the gut through this transporter, calcium channel blockers will enhance renal urinary iron excretion and iron chelators can bind and excrete iron that is already within the body.
The combination therapies of the invention may reduce the number or frequency of phlebotomies or therapeutic erythrocytapheresis treatments required to meet hemochromatosis patient serum iron and tissue iron therapeutic targets. The combination therapies of the invention may also prevent iron overload in patients or may prevent re-loading of iron overlaod in patients that have concluded their phlebotomy treatments.
The general value of the combination therapies of the invention in decreasing serum iron levels, can be determined using the assays described herein or below in the Biological Assays section. Alternatively, the general value of the combination therapies of the invention in treating iron overload disorders in mammals, preferably humans, may be established in industry standard animal models for demonstrating the efficacy of combination therapies in treating iron overload disorders.
For example, an iron deficient anemia, and therefore a hyperabsorptive iron state, may be induced by dietary means, or by treatment with phenylhydrazine, or by phlebotomy (Refino et al., Am. J. Clin. Nutr. 1983, 37:904-909; Redondo et al., Lab. Animal Sci. 1995, 45:578-583; Frazer ef a/., Gastroenterology, 2002, 123:835-844). Alternatively, iron absorption can also be stimulated by creating an hypoxic state to stimulate erythropoiesis (Raja et al., Br. J. Haematol., 1988, 68:373-378). In these models, combination therapies of the invention can be assessed by measuring reduced iron flux via the duodenum acutely or by monitoring whether chronic exposure to a combination therapy causes a decrease in the amount of iron loading as measured by serum iron, transferrin saturation, ferritin and liver iron. Alternatively, iron flux in these animals can be measured by tracing the absorption of radioactive iron administered orally. These experiments can also be performed in iron replete animals, although changes in these parameters will be less pronounced and therefore compound efficacy will be more difficult to judge.
Alternatively, genetic rat models of iron overload offer another format to show efficacy of the combination therapies of the invention. These models are applicable to a variety of iron disorders such as hereditary hemochromatosis (Levy et al., Blood, 1999, 94:9-11 ), juvenile hemochromatosis (Huang et al., J. Clin. Invest, 2005 115:2187-2191), beta-2-microglobulin (de Sousa et al., Immun. Lett, 1994, 39:105- 111 ), thalassemia (Ciavatta et al., Proc. Nat. Acad. Sci., 1995, 92: 9259-9263), hypotransferrinmia (Craven et al., Proc. Nat. Acad. ScL, 1987, 84(10):3457-61 ) and other hypochromic microcytic anemias. A combination therapy's efficacy can be assessed by measuring reduced iron flux via the duodenum acutely or by monitoring whether chronic exposure to a compound causes a decrease in the amount of iron loading as judged by serum iron, transferrin saturation, ferritin and liver iron. Alternatively, iron flux in these animals can be measured by tracing the absorption of radioactive iron administered orally.
Typically, a successful active ingredient of the present invention, including the DMT1 inhibitors, will meet some or all of the following criteria. Oral availability should be at less than 5%. Animal model efficacy is less than about 0.1 μg to about 100 mg/Kg body weight and the target human dose is between 0.1 μg to about 100 mg/Kg body weight, although doses outside of this range may be acceptable ("mg/Kg" means milligrams of compound per kilogram of body mass of the subject to whom it is being administered). The potency (as expressed by IC50 value) should be less than 10 μM, preferably below 1 μM and most preferably below 50 nM. The IC50 ("Inhibitory Concentration - 50%") is a measure of the amount of compound required to achieve 50% inhibition of DMT1 , over a specific time period, in an assay of the invention. ADMINISTRATION OF THE COMBINATION THERAPY OF THE INVENTION
Administration of the active ingredients of the combination therapy of the invention, in pure form or in an appropriate pharmaceutical composition, to a mammal in need thereof, can be carried out via any of the accepted modes of administration of agents for serving similar utilities. The recipients of the combination therapy of the invention can be any vertebrate animal, such as mammals. Among mammals, the preferred recipients are mammals of the Orders Primate (including humans, apes and monkeys), Arteriodactyla (including horses, goats, cows, sheep, pigs), Rodenta (including mice, rats, rabbits, and hamsters), and Camivora (including cats, and dogs). Among birds, the preferred recipients are turkeys, chickens and other members of the same order. The most preferred recipients are humans.
Each active ingredient, or a pharmaceutical composition comprising a therapeutically effective amount of the active ingredient, may be concomitantly administered to a mammal in need thereof. The term "concomitantly administered" means the administration of the active ingredients substantially concurrently. The term "concomitantly administered" encompasses not only administering the active ingredients in a single pharmaceutical dosage form but also the administration of each active ingredient in its own pharmaceutical dosage form, such as a pharmaceutical composition for oral administration. Alternatively, each active ingredient, or a pharmaceutical composition comprising a therapeutically effective amount of the active ingredient, may be sequentially administered to a mammal in need thereof. The term "sequentially administered" means the administration of the active ingredients at separately staggered times. Thus the active ingredient can be sequentially administered such that the beneficial pharmaceutical effects of the active ingredients are realized by the mammal at substantially the same time, depending on the pharmaceutically active half-life of each active ingredient.
The active ingredients of the combination therapy of the invention are administered in a therapeutically effective amount, which will vary depending upon a variety of factors including the activity of the specific active ingredient employed; the metabolic stability and length of action of the active ingredient; the age, body weight, general health, sex, and diet of the patient; the mode and time of administration; the rate of excretion; the drug combination; the severity of the particular disorder or condition; and the subject undergoing therapy.
The preferred daily dosage amount of each active ingredient will be tailored to the individual subject, as is understood and determinable by one skilled in the relevant arts, (see, e.g., Physicians' Desk Reference (latest edition); Berkowet al., eds., The Merck Manual, 16th edition, Merck and Co., Rahway, N.J., 1992; Goodman et al., eds., Goodman and Gilman's The Pharmacological Basis of Therapeutics, 11th edition, The MacGraw-Hill Companies, (2005); Avery's Drug Treatment: Principles and Practice of Clinical Pharmacology and Therapeutics, 3rd edition, ADIS Press, LTD., Williams and Wilkins, Baltimore, MD. (1987), Ebadi, Pharmacology, Little, Brown and Co., Boston, (1985); Osolci al., eds., Remington's Pharmaceutical Sciences, 18th edition, Mack Publishing Co., Easton, PA (1990); Katzung, Basic and Clinical Pharmacology, Appleton and Lange, Norwalk, CT (1992)). The ranges of therapeutically effective daily dosages for each active ingredient provided herein are therefore not intended to be limiting.
Generally, treatment with a combination therapy of the invention may be initiated using a daily dosage amount of each active ingredient which is less than the optimum daily dosage amount of the active ingredient. Thereafter, the dosage may be increased by small increments until the optimum effect under the circumstances is reached, i.e., until a negative iron balance in the mammal undergoing treatment.
Generally, a therapeutically effective daily dosage amount of a DMT1 inhibitor of the combination therapies of the invention is from about 0.001 mg/Kg body weight to about 100 mg/Kg body weight; preferably a therapeutically effective dose is from about 0.01 mg/Kg body weight to about 75 mg/Kg body weight; more preferably a therapeutically effective dose is from about 1 mg/Kg body weight to about 50 mg/Kg body weight. In preferred embodiments, a therapeutically effective daily dosage amount of a DMT1 inhibitor is daily dosage amount is 10 mg/Kg body weight, 25 mg/Kg body weight, or 50 mg/Kg body weight. The DMT1 inhibitor may be dosed orally once, twice, three times or four or more times a day.
Generally, a therapeutically effective daily dosage amount of a pharmaceutically acceptable iron chelator of the combination therapies of the invention is from about 10 mg/Kg body weight to about 100 mg/Kg body weight. In preferred embodiments, a therapeutically effective daily dosage amount of a pharmaceutically acceptable iron chelator of the combination therapies of the invention is about 10 mg/Kg body weight , about 20 mg/Kg body weight , about 30 mg/Kg body weight , about 50 mg/Kg body weight , about 75 mg/Kg body weight or about 100 mg/Kg body weight. The pharmaceutically acceptable iron chelator can be orally administered one, two, three or more times daily depending on the daily dosage. Alternatively, pharmaceutically acceptable iron chelators can be intravenously infused daily for up to 12 hours. In other embodiments, the pharmaceutically acceptable iron chelators can be administered by subcutaneous infusion in therapeutically effective dosage in treating iron overload. Generally, a therapeutically effective daily dosage amount of a PPI of the combination therapies of the invention is from about 1 mg to about 120 mg. In preferred embodiments, a therapeutically effective daily dosage of a PPI of the combination therapies of the invention is about 1 mg/Kg body weight, about 5 mg/Kg body weight, about 10 mg/Kg body weight, about 15 mg/Kg body weight, about 20 mg/Kg body weight, about 30 mg/Kg body weight, about 40 mg/Kg body weight, about 60 mg/Kg body weight, about 90 mg/Kg body weight, about 100 mg/Kg body weight, or about 120 mg/Kg body weight. Preferably, the daily dosage amount per body weight is 15 mg/Kg, 40 mg/Kg or 75 mg/Kg. The daily dosage amount of a PPI in a combination therapy of the invention can be less than what the current recommended daily dosage is for the PPI. PPIs can be administered orally once, twice or three times a day depending on the dosage. For example, Esomeprazol (Nexium) can be administered orally once a day to adults at 20 mg or 40 mg dosage, or at 10 mg or 20 mg dosage to children 1 to 11 years, or 20 mg or 40 mg dosage to children 12 to 17 years.
Generally, a therapeutically effective daily dosage amount of a calcium channel blocker of the combination therapies of the invention is from about 0.001 mg/Kg body weight to about 100 mg/Kg body weight; preferably a therapeutically effective dose is from about 0.01 mg/Kg body weight to about 75 mg/Kg body weight; more preferably a therapeutically effective dose is from about 0.1 mg/Kg body weight to about 25 mg/Kg body weight. In a preferred embodiment, a therapeutically effective daily dosage amount of a calcium channel blocker is about 0.1 mg/Kg body weight, about 1.0 mg/Kg body weight, or about 10 mg/Kg body weight. In another preferred embodiment, a therapeutically effective daily dosage amount of a calcium channel blocker is about 0.1 mg/Kg body weight, about 2.0 mg/Kg body weight, or about 20 mg/Kg body weight. The daily dosage amount of a calcium channel blocker in a combination therapy of the invention can be less that what the current recommended daily dosage is for a particular calcium channel blocker. The calcium channel blocker may be dosed orally once, twice, three times or four or more times a day. For example, the recommended starting dosage for immediate-release nifedipine capsules is 10 mg, taken three times daily and the recommended starting dosage for extended-release nifedipine is 30 to 60 mg, taken once daily. . Each active ingredient of the combination therapies of the invention may be dosed as if it is the only therapy being taken alone as each active ingredient may work additively through different mechanisms. Combination therapy may be from 1 month to 12 months or from 1 year to 5 years or for the life of the recipient. In another aspect, the combination therapies of the invention may be administered to the mammal, preferably the human, in combination with phlebotomy treatment. Therapeutic phlebotomy entails periodic removal of fixed amounts of blood. Typically, phlebotomy will remove about 250 mg of iron with each treatment per week. The combination therapies maybe administered prior to the initiation of phlebotomy, during phlebotomy therapy, after phlebotomy therapy has concluded or at all of these timepoints.
Pharmaceutical compositions comprising a therapeutically effective amount of a DMT1 inhibitor and a pharmaceutically acceptable excipient are disclosed in PCT Published Application No. WO 2008/109840; PCT Published Application No. WO 2008/115999; PCT Published Application No. WO 2008/118790; PCT Published Application No. WO 2008/121861 ; and PCT Published Application No. WO 2008/151288; the disclosures of which are incorporated in full by reference herein. Pharmaceutical compositions for pharmaceutically acceptable iron chelators, proton pump inhibitors and calcium channel blockers are well-known in the art. The present invention also relates to pharmaceutical compositions (hereinafter referred to as "combination-therapy pharmaceutical compositions") comprising a therapeutically effective amount of a DMT1 inhibitor and a therapeutically effective amount of a pharmaceutically acceptable iron chelator and/or a therapeutically effective amount of a proton pump inhibitor, and/or a therapeutically effective amount of a calcium channel blocker, and a pharmaceutically acceptable excipient.
Such combination-therapy pharmaceutical compositions can be prepared by combining the active ingredients with an appropriate pharmaceutically acceptable carrier, diluent or excipient, and may be formulated into preparations in solid, semi-solid, liquid or gaseous forms, such as tablets, capsules, powders, granules, ointments, solutions, suppositories, injections, inhalants, gels, microspheres, and aerosols. Typical routes of administering such combination-therapy pharmaceutical compositions include, without limitation, oral, topical, transdermal, inhalation, parenteral, sublingual, rectal, vaginal, and intranasal. The term parenteral as used herein includes subcutaneous injections, intravenous, intramuscular, intrasternal injection or infusion techniques. Such combination-therapy pharmaceutical compositions are formulated so as to allow the active ingredients contained therein to be bioavailable upon administration of the composition to a patient.
The combination-therapy pharmaceutical compositions disclosed herein also contain a pharmaceutically acceptable carrier, including any suitable diluent or excipient, which includes any pharmaceutical agent that does not itself induce the production of antibodies harmful to the individual receiving the composition, and which may be administered without undue toxicity. Pharmaceutically acceptable carriers include, but are not limited to, liquids, such as water, saline, glycerol and ethanol, and the like. A thorough discussion of pharmaceutically acceptable carriers, diluents, and other excipients is presented in REMINGTON'S PHARMACEUTICAL SCIENCES (Mack Pub. Co., N.J. current edition).
The combination-therapy pharmaceutical compositions disclosed herein may be in the form of a solid or liquid. In one aspect, the carrier(s) are particulate, so that the compositions are, for example, in tablet or powder form. The carrier(s) may be liquid, with the compositions being, for example, an oral syrup, injectable liquid or an aerosol, which is useful in, for example, inhalatory administration.
When intended for oral administration, the combination-therapy pharmaceutical compositions disclosed herein is preferably in either solid or liquid form, where semi-solid, semi-liquid, suspension and gel forms are included within the forms considered herein as either solid or liquid.
As a solid composition for oral administration, the combination-therapy pharmaceutical compositions disclosed herein may be formulated into a powder, granule, compressed tablet, pill, capsule, chewing gum, wafer or the like form. Such a solid composition will typically contain one or more inert diluents or edible carriers. In addition, one or more of the following may be present: binders such as carboxymethylcellulose, ethyl cellulose, microcrystalline cellulose, gum tragacanth or gelatin; excipients such as starch, lactose or dextrins, disintegrating agents such as alginic acid, sodium alginate, Primogel, corn starch and the like; lubricants such as magnesium stearate or Sterotex; glidants such as colloidal silicon dioxide; sweetening agents such as sucrose or saccharin; a flavoring agent such as peppermint, methyl salicylate or orange flavoring; and a coloring agent.
When the combination-therapy pharmaceutical compositions disclosed herein are in the form of capsules, for example, a gelatin capsule, they may contain, in addition to materials of the above type, a liquid carrier such as polyethylene glycol or oil. The combination-therapy pharmaceutical compositions disclosed herein may be in the form of a liquid, for example, an elixir, syrup, solution, emulsion or suspension. The liquid may be for oral administration or for delivery by injection, as two examples. When intended for oral administration, preferred composition contain, in addition to the active ingredients, one or more of a sweetening agent, preservatives, dye/colorant and flavor enhancer. In a composition intended to be administered by injection, one or more of a surfactant, preservative, wetting agent, dispersing agent, suspending agent, buffer, stabilizer and isotonic agent may be included.
Liquid combination-therapy pharmaceutical compositions, whether they be solutions, suspensions or other like form, may include one or more of the following adjuvants: sterile diluents such as water for injection, saline solution, preferably physiological saline, Ringer's solution, isotonic sodium chloride, fixed oils such as synthetic mono or diglycerides which may serve as the solvent or suspending medium, polyethylene glycols, glycerin, propylene glycol or other solvents; antibacterial agents such as benzyl alcohol or methyl paraben; antioxidants such as ascorbic acid or sodium bisulfite; chelating agents such as ethylenediaminetetraacetic acid; buffers such as acetates, citrates or phosphates and agents for the adjustment of tonicity such as sodium chloride or dextrose. The parenteral preparation can be enclosed in ampoules, disposable syringes or multiple dose vials made of glass or plastic. Physiological saline is a preferred adjuvant. An injectable pharmaceutical composition is preferably sterile.
A liquid combination-therapy pharmaceutical composition of the invention intended for either parenteral or oral administration should contain an amount of a compound of the invention such that a suitable dosage will be obtained. Typically, this amount is at least 0.01 % of a compound of the invention in the composition. When intended for oral administration, this amount may be varied to be between 0.1 and about 70% of the weight of the composition. Preferred oral combination-therapy pharmaceutical compositions contain between about 4% and about 50% of the compound of the invention. Preferred combination-therapy pharmaceutical compositions and preparations according to the present invention are prepared so that a parenteral dosage unit contains between 0.01 to 10% by weight of the compound prior to dilution of the invention.
The combination-therapy pharmaceutical composition of the invention may be intended for topical administration, in which case the carrier may suitably comprise a solution, emulsion, ointment or gel base. The base, for example, may comprise one or more of the following: petrolatum, lanolin, polyethylene glycols, bee wax, mineral oil, diluents such as water and alcohol, and emulsifiers and stabilizers. Thickening agents may be present in a pharmaceutical composition for topical administration. If intended for transdermal administration, the composition may include a transdermal patch or iontophoresis device. Topical formulations may contain a concentration of the compound of the invention from about 0.1 to about 10% w/v (weight per unit volume). The combination-therapy pharmaceutical composition of the invention may be intended for rectal administration, in the form, for example, of a suppository, which will melt in the rectum and release the drug. The composition for rectal administration may contain an oleaginous base as a suitable nonirritating excipient. Such bases include, without limitation, lanolin, cocoa butter and polyethylene glycol.
The combination-therapy pharmaceutical composition of the invention may include various materials, which modify the physical form of a solid or liquid dosage unit. For example, the composition may include materials that form a coating shell around the active ingredients. The materials that form the coating shell are typically inert, and may be selected from, for example, sugar, shellac, and other enteric coating agents. Alternatively, the active ingredients may be encased in a gelatin capsule.
The combination-therapy pharmaceutical composition of the invention in solid or liquid form may include an agent that binds to the compound of the invention and thereby assists in the delivery of the compound. Suitable agents that may act in this capacity include a monoclonal or polyclonal antibody, a protein or a liposome.
The combination-therapy pharmaceutical composition of the invention may consist of dosage units that can be administered as an aerosol. The term aerosol is used to denote a variety of systems ranging from those of colloidal nature to systems consisting of pressurized packages. Delivery may be by a liquefied or compressed gas or by a suitable pump system that dispenses the active ingredients. Aerosols of compounds of the invention may be delivered in single phase, bi-phasic, or th-phasic systems in order to deliver the active ingredient(s). Delivery of the aerosol includes the necessary container, activators, valves, subcontainers, and the like, which together may form a kit. One skilled in the art, without undue experimentation may determine preferred aerosols.
The combination-therapy pharmaceutical compositions of the invention may be prepared by methodology well known in the pharmaceutical art. For example, a combination-therapy pharmaceutical composition intended to be administered by injection can be prepared by combining a compound of the invention with sterile, distilled water so as to form a solution. A surfactant may be added to facilitate the formation of a homogeneous solution or suspension. Surfactants are compounds that non-covalently interact with the compound of the invention so as to facilitate dissolution or homogeneous suspension of the compound in the aqueous delivery system. The combination-therapy pharmaceutical compositions of the invention can be formulated so as to provide quick, sustained or delayed release of the active ingredient after administration to the patient by employing procedures known in the art. Controlled release drug delivery systems include osmotic pump systems and dissolutional systems containing polymer-coated reservoirs or drug-polymer matrix formulations. Examples of controlled release systems are given in U.S. Pat. Nos. 3,845,770 and 4,326,525 and in P. J. Kuzma et al, Regional Anesthesia 22 (6): 543- 551 (1997), all of which are incorporated herein by reference.
The combination-therapy pharmaceutical compositions of the invention can also be delivered through intra-nasal drug delivery systems for local, systemic, and nose-to-brain medical therapies. Controlled Particle Dispersion (CPD)™ technology, traditional nasal spray bottles, inhalers or nebulizers are known by those skilled in the art to provide effective local and systemic delivery of drugs by targeting the olfactory region and paranasal sinuses.
Current methods for ocular delivery include topical administration (eye drops), subconjunctival injections, periocular injections, intravitreal injections, surgical implants and iontophoresis (uses a small electrical current to transport ionized drugs into and through body tissues). Those skilled in the art would combine the best suited excipients with the compound for safe and effective intra-occular administration.
BIOLOGICAL ASSAYS Various techniques are known in the art for testing the effectiveness of the combination therapies of the invention. In order that the invention described herein may be more fully understood, the following biological assays are set forth. It should be understood that these examples are for illustrative purposes only and are not to be construed as limiting this invention in any manner.
BIOLOGICAL ASSAY 1
The Effect of a Combination Therapy on the Acute Iron Deficiency Rat Model
This assay was conducted to determine if a single oral dose of 75 tng/Kg Lansoprazole or 50 mg/Kg of Compound A (i.e. a DMT1 inhibitor) can block the observed increase of serum iron in a dose dependent manner in a previously developed model of iron deficiency in Sprague Dawley rats when a 1 mg/Kg oral iron challenge is given 2 hours following drug dosing. This assay was also conducted to determine if a combination therapy consisting of an oral dose of 15 mg/Kg Lansoprazole and 10 mg/Kg Compound A or an oral dose of 40 mg/Kg Lansoprazole and 25 mg/Kg Compound A can block the observed increase of serum iron in a dose dependent manner in a previously developed model of iron deficiency in Sprague Dawley rats when a 1 mg/Kg oral iron challenge is given 2 hours following dosing of the active ingredients. Methods
Four-week-old Sprague Dawley rats were placed on an iron deficient diet (TD.80396) for 3 weeks. Twenty-seven rats were allocated to one of the treatment groups (N = 3 or 6) ensuring that weights were not statistically different between the groups. An additional four animals were utilized to establish baseline serum iron levels. Lansoprazole was administered twice at 18 and 2 hours before the iron challenge. Compound A was dosed once orally following a four-hour fast. Animals were then given an oral iron challenge two hours later. The iron challenge consisted of an oral dose of elemental iron at 1 mg/Kg. The rats' individual iron status was evaluated by measuring serum iron 1 hour following iron challenge as an endpoint. Test compounds, Vehicles and Iron Challenge
1. Lansoprazole (PPI)
Vehicle: 0.2% Tween20 : 99.8% CMC (carboxymethylcellulose) (1%) adjusted to pH 9.0. Dose volume: 4 mL/Kg. Dose: 15, 40 or 75 mg/Kg.
2. Compound A (Cpd A)
Vehicle: 5% DMSO : 10% EtOH : 30% PEG400 : 35% PG : 20% H2O.
Dose volume: 4 mL/Kg. Dose: 10, 25 or 50 mg/Kg. 3. Iron Challenge
Iron solutions were made up fresh just prior to iron challenge to prevent excessive oxidation. FeSO4 «7H2O crystals were flushed with nitrogen following use to prevent oxidation. 0.25 mg/mL FeSO4*7H2O was formulated as 0.325 g in 50 mL H2O.
Dose volume: 1 mL/Kg (1 mg/Kg of FeSO4)
Results
The iron-deficient rats responded to the oral iron challenge by absorbing iron rapidly, such that the vehicle for Lansoprazole and the vehicle for Compound A both resulted in an 11 -fold increase in serum iron within 1 hour of the iron challenge.
As shown below in Table 1 , oral administration of 75 mg/Kg Lansoprazole (PPI) alone blocked iron absorption by approximately 56% when compared to the oral administration of the vehicle alone. Oral administration of 50 mg/Kg Compound A (Cpd A) alone blocked iron absorption by approximately 62% when compared to the oral administration of the vehicle alone.
Oral administration of a combination of 15 mg/Kg Lansoprazole and 10 mg/Kg Compound A blocked iron absorption by approximately 33% when compared to the oral administration of the appropriate vehicle alone.
Oral administration of a combination of 40 mg/Kg Lansoprazole and 25 mg/Kg Compound A blocked iron absorption by approximately 75% when compared to the oral administration of the appropriate vehicle alone.
Administration of each of the combination therapies appears to have resulted in a higher blocked iron absorption percentage than when the active ingredients therein are administered at a higher dose alone.
Table 1 : Serum Iron Level Change
Figure imgf000065_0001
* PPI = Lansoprazole ** Cpd A = Compound A BIOLOGICAL ASSAY 2
The Effect of a Combination Therapy on the Acute Iron Deficiency Rat Model This assay was conducted to determine if a single oral dose of 15, 40 or 75 mg/Kg Lansoprazole or 10, 25 or 50 mg/Kg of Compound A (i.e. a DMT1 inhibitor) can block the observed increase of serum iron in a dose dependent manner in a previously developed model of iron deficiency in Sprague Dawley rats when a 1 mg/Kg oral iron challenge is given 2 hours following drug dosing. This assay was also conducted to determine if a combination therapy consisting of an oral dose of 15 mg/Kg Lansoprazole and 10 mg/Kg Compound A, an oral dose of 40 mg/Kg Lansoprazole and 25 mg/Kg Compound A, or an oral dose of 75 mg/Kg Lansoprazole and 50 mg/Kg Compound A can block the observed increase of serum iron in a dose dependent manner in a previously developed model of iron deficiency in Sprague Dawley rats when a 1 mg/Kg oral iron challenge is given 2 hours following dosing of the active ingredients. Methods
Four-week-old Sprague Dawley rats were placed on an iron deficient diet (TD.80396) for 3 weeks. Forty-four rats were allocated to one of the treatment groups (N = 4) ensuring that weights were not statistically different between the groups. An additional four animals were utilized to establish baseline serum iron levels. Lansoprazole was administered twice at 18 and 2 hours before the iron challenge. Compound A was dosed once orally following a four-hour fast. Animals were then given an oral iron challenge two hours later. The iron challenge consisted of an oral dose of elemental iron at 1 mg/Kg. The rats' individual iron status was evaluated by measuring serum iron 1 hour following iron challenge as an endpoint. Test compounds, Vehicles and Iron Challenge
1. Lansoprazole
Vehicle: 0.2% Tween20 : 99.8% CMC (carboxymethylcellulose) (1%) adjusted to pH 9.0. Dose volume: 4 mL/Kg. Dose: 15, 40 or 75 mg/Kg.
2. Compound A
Vehicle: 5% DMSO : 10% EtOH : 30% PEG400 : 35% PG : 20% H2O.
Dose volume: 4 mL/Kg. Dose: 10, 25 or 50 mg/Kg. 3. Iron Challenge
Iron solutions were made up fresh just prior to iron challenge to prevent excessive oxidation. FeSO4*7H2O crystals were flushed with nitrogen following use to prevent oxidation. 0.25 mg/mL FeSO4*7H2O was formulated as 0.325 g in 50 ml_ H2O. Dose volume: 1 mL/Kg (1 mg/Kg of FeSO4)
Results
The iron-deficient rats responded to the oral iron challenge by absorbing iron rapidly, such that the vehicle for Lansoprazole and the vehicle for Compound A both resulted in an 4-fold increase in serum iron within 1 hour of the iron challenge.
As shown below in Tables 2a, oral administration of 15 mg/Kg Lansoprazole alone blocked iron absorption by approximately 4% when compared to the oral administration of the vehicle alone. Oral administration of 40 mg/Kg Lansoprazole alone blocked iron absorption by approximately 27% when compared to the oral administration of the vehicle alone.
Oral administration of 75 mg/Kg Lansoprazole alone blocked iron absorption by approximately 38% when compared to the oral administration of the vehicle alone.
Oral administration of 10 mg/Kg Compound A alone blocked iron absorption by approximately 16% when compared to the oral administration of the vehicle alone.
Oral administration of 25 mg/Kg Compound A alone blocked iron absorption by approximately 11 % when compared to the oral administration of the vehicle alone.
Oral administration of 50 mg/Kg Compound A alone blocked iron absorption by approximately 89% when compared to the oral administration of the vehicle alone. As shown below in Table 2b, oral administration of a combination of 15 mg/Kg
Lansoprazole and 10 mg/Kg Compound A blocked iron absorption by approximately 27% when compared to the oral administration of the appropriate vehicle alone.
Oral administration of a combination of 40 mg/Kg Lansoprazole and 25 mg/Kg Compound A blocked iron absorption by approximately 70% when compared to the oral administration of the appropriate vehicle alone.
Oral administration of a combination of 75 mg/Kg Lansoprazole and 50 mg/Kg Compound A blocked iron absorption by approximately 97% when compared to the oral administration of the appropriate vehicle alone.
Administration of each of the combination therapies appears to have resulted in a higher blocked iron absorption percentage than when the active ingredients therein are administered alone.
Table 2a: Serum Iron Level Change (Individual Active Ingredients)
Figure imgf000068_0001
PPl = Lansoprazole * Cpd A = Compound A
Table 2b: Serum Iron Level Change (Combination Therapies)
Figure imgf000068_0002
* PPI = Lansoprazole
** Cpd A = Compound A
1 an average value for vehicle was calculated form the values of the two vehicle's data.
BIOLOGICAL ASSAY 3 The Effect of a Combination Therapy on the Acute Iron Deficiency Rat Model
This assay was conducted to determine if a single oral dose of 75 mg/Kg Omeprazole or 50 mg/Kg of Compound A (i.e. a DMT1 inhibitor) can block the observed increase of serum iron in a dose dependent manner in a previously developed model of iron deficiency in Sprague Dawley rats when a 1 mg/Kg oral iron challenge is given 2 hours following drug dosing. This assay was also conducted to determine if a combination therapy consisting of an oral dose of 40 mg/Kg Omeprazole and 25 mg/Kg Compound A can block the observed increase of serum iron in a dose dependent manner in a previously developed model of iron deficiency in Sprague Dawley rats when a 1 mg/Kg oral iron challenge is given 2 hours following dosing of the active ingredients.
Methods Four-week-old Sprague Dawley rats were placed on an iron deficient diet
(TD.80396) for 3 weeks. Eighteen rats were allocated to one of the treatment groups (N = 2 or 4) ensuring that weights were not statistically different between the groups. An additional four animals were utilized to establish baseline serum iron levels. Omeprazole was administered twice at 18 and 2 hours before the iron challenge. Compound A was dosed once orally following a four-hour fast. Animals were then given an oral iron challenge two hours later. The iron challenge consisted of an oral dose of elemental iron at 1 mg/Kg. The rats' individual iron status was evaluated by measuring serum iron 1 hour following iron challenge as an endpoint. Baseline serum iron levels were not taken. Therefore termination serum iron levels were provided instead of change in serum iron levels. Only one vehicle group (the vehicle for Omeprazole) was included due to the limited number of available animals.
Test compounds, Vehicles and Iron Challenge
1. Omeprazole
Vehicle: 0.2% Tween20 : 99.8% CMC (carboxymethylcellulose) (1%) adjusted to pH 9.0.
Dose volume: 4 mL/Kg. Dose: 40 or 75 mg/Kg.
2. Compound A
Vehicle: 5% DMSO : 10% EtOH : 30% PEG400 : 35% PG : 20% H2O. Dose volume: 4 mL/Kg.
Dose: 25 or 50 mg/Kg.
3. Iron Challenge
Iron solutions were made up fresh just prior to iron challenge to prevent excessive oxidation. FeSO4*7H2O crystals were flushed with nitrogen following use to prevent oxidation. 0.25 mg/mL FeSO4*7H2O was formulated as 0.325 g in 50 ml_ H2O. Dose volume: 1 mL/Kg (1 mg/Kg of FeSO4)
Result and Conclusion
As shown below in Table 3, oral administration of a combination of 40 mg/Kg Omeprazole and 25 mg/Kg Compound A blocked iron absorption by approximately 69% when compared to the oral administration of the Omeprazole vehicle.
Oral administration of a combination of 75 mg/Kg Omeprazole and 50 mg/Kg Compound A blocked iron absorption by approximately 76% when compared to the oral administration of the Omeprazole vehicle.
Oral administration of only 75 mg/Kg Omeprazole blocked iron absorption by approximately 52% when compared to the oral administration of only the vehicle.
Oral administration of only 50 mg/Kg Compound A blocked iron absorption by approximately 55% when compared to the oral administration of only the vehicle.
Administration of each of the combination therapies appears to have resulted in a higher blocked iron absorption percentage than when the active ingredients therein are administered alone.
Table 3: Termination Serum Iron Levels
Figure imgf000070_0001
* * * * * All of the U.S. patents, U.S. patent application publications, U.S. patent applications, foreign patents, foreign patent applications and non-patent publications referred to in this specification are incorporated herein by reference in their entireties.
Although the foregoing invention has been described in some detail to facilitate understanding, it will be apparent that certain changes and modifications may be practiced within the scope of the appended claims. Accordingly, the described embodiments are to be considered as illustrative and not restrictive, and the invention is not to be limited to the details given herein, but may be modified within the scope and equivalents of the appended claims.

Claims

WHAT IS CLAIMED IS
1. A method of treating an iron overload disorder in a mammal, wherein the method comprises administering to the mammal in need thereof a therapeutically effective amount of a DMT1 inhibitor and a therapeutically effective amount of a pharmaceutically acceptable iron chelator.
2. The method of Claim 1 wherein the iron chelator is selected from Deferasirox, Deferiprone, Deferitrin or Deferoxamine.
3. The method of Claims 1 or 2 further comprising the administration of a therapeutically effective amount of a proton pump inhibitor.
4. A method of treating an iron overload disorder in a mammal, wherein the method comprises administering to the mammal in need thereof a therapeutically effective amount of a DMT1 inhibitor and a therapeutically effective amount of a proton pump inhibitor.
5. The method of any one of Claims 3 to 4 wherein the proton pump inhibitor is selected from Omeprazole, Esomeprazole, Lansoprazole, Pantoprazole, Rabeprazole or Tenatoprazole.
6. The method of Claim 5 wherein the proton pump inhibitor is selected from Omeprazole or Lansoprazole.
7. The method of any one of Claims 1 to 6 wherein the DMT1 inhibitor is selected from:
(2,4,6-trimethyl-1 ,3-phenylene)bis(methylene) dicarbamimidothioate; dibenzo[b,c(]thiophene-4,6-diylbis(methylene) dicarbamimidothioate; (2-fluorodibenzo[/b,d]thiophene-4,6-diyl)bis(methylene) dicarbamimidothioate; (3,7-dibromodibenzo[t»,c/]thiophene-4,6-diyl)bis(methylene) dicarbamimidothioate; (2-chloro-8-fluorodibenzo[ib,cf]thiophene-4,6-diyl)bis(methylene) dicarbamimidothioate; (3-bromodibenzo[b,c/]thiophene-4,6-diyl)bis(methylene) dicarbamimidothioate; dibenzo[b,c/]thiophene-1 ,9-diylbis(methylene) dicarbamimidothioate; (9-oxo-9/-/-xanthene-4,5-diyl)bis(methylene) dicarbamimidothioate; dibenzo[jb,d]furan-4,6-diylbis(methylene) dicarbamimidothioate; (3, 7-dimethy!dibenzo[b,cf]furan-4,6-diyl)bis(methylene) dicarbamimidothioate; (3,7-dichloroldibenzo[b,c(]furan-4,6-diyl)bis(methylene) dicarbamimidothioate; (3,7-dibromodibenzo[jfc),c(]furan-4,6-diyl)bis(methylene) dicarbamimidothioate; (2-fluorodibenzo[jb,cdfuran-4,6-diyl)bis(methylene) dicarbamimidothioate; (2,8-dibromodibenzo[b,c/]furan-4,6-diyl)bis(methylene) dicarbamimidothioate; biphenylene-1 ,8-diylbis(methylene) dicarbamimidothioate; (3,6-difluorobiphenylene-1 ,8-diyl)bis(methylene) dicarbamimidothioate; biphenylene-1 , 4, 5, δ-tetrayltetrakis(methylene) tetracarbamimidothioate; 2-(8-carbamimidoylsulfanylmethyl-9-oxo-9H-fluoren-1-ylmethyl)-isothiourea; (9-oxo-9/-/-fluorene-4,5-diyl)bis(methylene) dicarbamimidothioate; 2-(2,7-di-tert-butyl-5-carbamimidoylsulfanylmethyl-9,9-dimethyl-9/-/-xanthen-4- ylmethyl)-isothiourea; phenoxathiine-4,6-diylbis(methylene) dicarbamimidothioate; (5,7-dihydrodibenzo[c,e]thiepine-1 ,1 i-diyl)bis(methylene) dicarbamimidothioate; 2-(2'-carbamimidoylsulfanyImethyl-biphenyl-2-ylmethyl)-isothiourea; (6,6'-dimethylbiphenyl-2,2'-diyl)bis(methylene) dicarbamimidothioate dihydrobromide; biphenyl-2, 2', 6, 6'-tetrayltetrakis(methylene) tetracarbamimidothioate; dimethyl 6,6'-bis(carbamimidoylthiomethyl)biphenyl-2,2'-dicarboxylate; 2-[2-(2-carbamimidoylsulfanylmethyl-phenoxy)-benzyl]-isothiourea; 2-(1-{2-[2-(1-carbamimidoylsulfanyl-ethyl)-phenoxy]-phenyl}-ethyl)-isothiourea; 2-[2-({[amino(imino)methyl]thio}methyl)-4-fluorophenoxy]-5-nitrobenzyl imidothiocarbamate; 2-[2-({[amino(imino)methyl]thio}methyl)-5-chlorophenoxy]-4-nitrobenzyl imidothiocarbamate; 2-[2-({[amino(imino)methyl]thio}methyl)-5-chlorophenoxy]-5-fluorobenzyl imidothiocarbamate;
2-[2-(2-carbamimidoylsulfanylmethyl-3-chlorophenoxy)-5-fluorobenzyl]isothiourea; 2-[2-(2-carbamimidoylsulfanylmethyl-4-chlorophenoxy)-5-fluorobenzyl]isothiourea; 2-[2-({[amino(imino)methyl]thio}methyl)-4-chlorophenoxy]benzyl imidothiocarbamate; 2-[2-({[amino(imino)methyl]thio}methyl)-3-chlorophenoxy]benzyl imidothiocarbamate; 2-[2-({[amino(imino)methyl]thio}methyl)-5-chlorophenoxy]benzyl imidothiocarbamate; 2-[2-({[amino(imino)methyl]thio}methyl)-5-chlorophenoxy]-5-nitrobenzyl imidothiocarbamate; -[2-({[amino(imino)methyl]thio}methyl)-4-chlorophenoxy]-5-nitrobenzyl imidothiocarbamate; -[2-(2-carbamimidoylsulfanylmethyl-5-fluorophenoxy)-5-fluorobenzyl]isothiourea;-[2-(2-carbamimidoylsulfanylmethyl-4-fluorophenoxy)-5-fluorobenzyl]isothiourea;-[2-({[amino(imino)methyl]thio}methyl)-4-fluorophenoxy]benzyl imidothiocarbamate;-[2-(2-carbamimidoylsulfanylmethyi-phenylsulfanyl)-benzyl]-isothiourea; -[2-(2-carbamimidoylsulfanylmethyl-4-fluoro-phenylsulfanyl)-5-fluoro-benzyl]- isothiourea; -[2-(2-carbamimidoylsulfanylmethyl-4-fluoro-phenylsulfanyl)-benzyl]-isothiourea;-[2-(2-carbamimidoylsulfanylmethyl-5-methyl-phenylsulfanyl)-benzyl]-isothiourea;-[2-(2-carbamimidoylsulfanylmethyl-4-methoxy-phenylsulfanyl)-benzyl]-isothiourea;-[2-(2-carbamimidoylsulfanylmethyl-5-methyl-phenylsulfanyl)-5-fluoro-benzyl]- isothiourea; -[2-(2-carbamimidoylsulfanylmethyl-4-methoxy-phenylsulfanyl)-5-fluoro-benzyl]- isothiourea; -[2-(2-carbamimidoylsulfanylmethyl-5-chloro-phenylsulfanyl)-5-fluoro-benzyl]- isothiourea; -[2-(2-carbamimidoylsulfanylmethyl-6-methylphenylsulfanyl)-benzyl]isothiourea;-[2-(2-carbamimidoylsulfanylmethyl-4,5-difluorophenylsulfanyl)-benzyl]isothiourea;-[2-(2-carbamimidoylsulfanylmethyl-4-methyl-phenyisulfanyl)-benzyl]-isothiourea;-[2-(2-carbamimidoylsulfanylmethyl-5-chloro-phenylsulfanyl)-benzyl]-isothiourea;-[2-(2-carbamimidoylsulfanylmethyl-5-fluoro-phenylsulfanyl)-benzyl]-isothiourea;-[2-(2-carbamimidoylsulfanylmethyl-5-fluoro-phenylsulfanyl)-5-fluorobenzyl]- isothiourea; -[2-(2-carbamimidoylsulfanylmethyl-4,5-difluorophenylsulfanyl)-5- fluorobenzyφsothiourea; -{[2-({[amino(imino)methyl]thio}methyl)-4-fluorophenyl]thio}-3-nitrobenzyl imidothiocarbamate; -{[2-({[amino(imino)methyl]thio}methyl)-4-fluorophenyl]thio}-5-nitrobenzyl imidothiocarbamate; -[2-(2-carbamimidoylsulfanylmethyl-4-fluorophenylsulfanyl)-5-
(trifluoromethyl)benzyl]isothiourea; -[2-(2-carbamimidoylsulfanylmethylphenylsulfanyl)-5-aminobenzyl]-isothiourea;-[2-(2-carbamimidoylsulfanylmethyl-4-chlorophenylsulfanyl)-5- fluorobenzyl]isothiourea; -[2-(2-carbamimidoylsulfanylmethylphenylsulfanyl)-5-ethylaminobenzyl]isothiourea;-[2-(2-carbamimidoylsulfanylmethyl-4-chlorophenylsulfanyl)-5- chlorobenzyl]isothiourea; -[2-(2-carbamimidoylsulfanylethyl-4-fluorophenylsulfanyl)-5-fluorobenzyl]isothiourea;-[2-(2-carbamimidoylsulfanylethyl-4-chlorophenylsulfanyl)-5-fluorobenzyl]isothiourea;-[2-(2-carbamimidoylsulfanylethylphenylsulfanyl)benzyl]isothiourea; -[2-(2-carbamimidoylsulfanylmethylphenylsulfanyl)-5-chlorobenzyl]-isothiourea; -[2-(2-carbamimidoylsulfanylmethylphenylsulfanyl)-5-
(trifluoromethyl)benzyl]isothiourea; -[2-(2-methylcarbamimidoylsulfanylmethylphenylsulfanyl)benzyl]-methylisothiourea;-[2-(2-carbamimidoylsulfanylmethyl-4-fluorophenylsulfanyl)-5-
(methylsulfonyl)benzyl]isothiourea; -({2-({[amino(imino)methyl]thio}methyl)-4-[(dimethylamino)sulfonyl]phenyl}thio)-5- fluorobenzyl imidothiocarbamate; -[2-(2-carbamimidoylsulfanylmethylphenylsulfanyl)benzyl]-isothiourea; -{[2-({[amino(imino)methyl]thio}methyl)-4-fluorophenyl]thio}-4-(methylsulfonyl)benzyl imidothiocarbamate; -{[2-({[amino(imino)methyl]thio}methyl)-4-chlorophenyl]thio}-5-cyanobenzyl imidothiocarbamate; -[2-(2-carbamimidoylsulfanylmethyiphenylsulfonyl)benzyl]-isothiourea; -[2-(2-carbamimidoylsulfanylmethyl-4-fluorophenylsulfonyl)-5-fluorobenzyl]isothiourea;-(6-((aminoamidino)thiomethyl)phenyl)thio-1-((aminoamidino)thiomethyl)benzene;-(6-(2-amidinoethyl)phenyl)thio-1-(2-amidinoethyl)benzene; -(6-(amidinomethyl)phenyl)thio-1-(amidinomethyl)benzene; -(6-(3-amidinopropyl)phenyl)thio-1-(3-amidinopropyl)benzene; -(6-((cyanoamidino)methyl)phenyl)thio-1-((cyanoamidino)methyl)benzene; -(6-(2-(cyanoamidino)ethyl)phenyl)thio-1-(2-(cyanoamidino)ethyl)benzene; -(6-(3-(cyanoamidino)propyl)phenyl)thio-1-(3-(cyanoamidino)propyl)benzene; -(2-(2-(guanidinomethyl)phenylthio)benzyl)guanidine; ,2'-thiobis(Λ/-(diaminomethylene)benzamide); -(6-(amidinothiomethyl)phenyl)carbonyl-1-(amidinothiomethyl)benzene; -(6-(amidinothiomethyl)phenyl)carbonyl-1-(amidinothiomethyl)-5-fluorobenzene; -(6-(amidinothiomethyl)phenyl)carbonyl-1-(amidinothiomethyl)-4-fluorobenzene; -(6-(amidinothiomethyl)phenyl)carbonyl-1-(amidinothiomethyl)-4-chlorobenzene; 2-(6-(amidinothiomethyl)phenyl)carbonyl-1-(amidinothiomethyl)-5-chlorobenzene; 4,4-diisothiourea benzophenone;
2,2-(methylazanediyl)bis(2,1-phenylene)bis(methylene)dicarbamimidothioate; 2-[2-(1-carbamimidoylsulfanylmethyl-naphthalen-2-ylsulfanyl)-benzyl]-isothiourea; 2-[2-(1-carbamimidoylsulfanylmethylnaphthalen-2-ylsulfanyl)-5-fluorobenzyl]- isothiourea; (2-{[2-({[amino(imino)methyl]thio}methyl)-4-fluoropheny!]thio}-3-thienyl)methyl imidothiocarbamate; (4-{[2-({[amino(imino)methyl]thio}methyl)-4-fluorophenyl]thio}-3-thienyl)methyl imidothiocarbamate;
2-pyridin-2-yl-4,5-dihydro-2/-/-benzo[e]indazol-1-ol; 7-bromo-2-pyridin-2-yl-4,5-dihydro-2H-benzo[e]indazol-1-ol; 2-pyridin-2-yl-2,4,5,6-tetrahydro-2,3-diaza-benzo[e]azulen-1-ol; 1-hydroxy-2-pyridin-2-y!-4,5-dihydro-2/-/-benzo[e]indazole-7-carbonitrile; 7-phenyl-2-pyridin-2-yl-4,5-dihydro-2/-/-benzo[e]indazol-1-ol; 7-benzo[1 ,3]dioxol-5-yl-2-pyridin-2-yl-4,5-dihydro-2/-/-benzo[e]indazol-1-ol; 4-(1-hydroxy-2-pyridin-2-yl-4,5-dihydro-2/-/-benzo[e]indazol-7-yl)-benzonitrile; 2-pyridin-2-yl-7-p-tolyl-4,5-dihydro-2H-benzo[e]indazol-1-ol; 7-(4-methoxyphenyl)-2-pyridin-2-yl-4,5-dihydro-2/-/-benzo[e]indazol-1-ol; 2-(1-hydroxy-4,5-dihydro-2/-/-benzo[e]indazol-2-yl)pyridine 1 -oxide; 7-methoxy-2-pyridin-2-yl-4,5-dihydro-2/-/-benzo[e]indazol-1-ol; 8-methoxy-2-pyridin-2-yl-4,5-dihydro-2/-/-benzo[e]indazol-1-ol; 2-(5-(trifluoromethyl)pyridin-2-yl)-4,5-dihydro-2/-/-benzo[e]indazol-1-ol; 2-(4-(trifluoromethyl)pyridin-2-yl)-4,5-dihydro-2/-/-benzo[e]indazol-1-ol; 2-(5-nitropyridin-2-yl)-4,5-dihydro-2H-benzo[e]indazol-1-ol; 2-(3-(trifluoromethyl)pyridin-2-yl)-4,5-dihydro-2H-benzo[e]indazol-1-ol; 2-(5-methylpyridin-2-yl)-4,5-dihydro-2/-/-benzo[e]indazol-1-ol; 2-(benzo[d]thiazol-2-yl)-4,5-dihydro-2/-/-benzo[e]indazol-1-ol; 2-(6-methoxybenzo[c/]thiazol-2-yl)-4,5-dihydro-2/-/-benzo[e]indazol-1-ol; 2-(6-fluorobenzo[c/]thiazol-2-yl)-4,5-dihydro-2/-/-benzo[e]indazol-1-ol; 2-(benzo[c/]thiazoI-2-yl)-7-methoxy-4,5-dihydro-2/-/-benzo[e]indazol-1-ol; 2-(6-methylbenzo[cdthiazol-2-yl)-4,5-dihydro-2H-benzo[e]indazol-1-ol; 2-(4-methylbenzo[d]thiazol-2-yl)-4,5-dihydro-2H-benzo[e]indazol-1-ol; 2-(1-hydroxy-4,5-dihydro-2H-benzo[e]indazol-2-yl)benzo[d]thiazole-6-carboxylic acid; 2-(1/-/-benzo[cdimidazol-2-yl)-4,5-dihydro-2/-/-benzo[e]indazol-1-ol; 2-(4-tert-butylthiazol-2-yl)-4,5-dihydro-2/-/-benzo[e]indazol-1-ol hydrochloride;
1 -(2-bromophenyl)-3-methyl-4-phenyl-1 /-/-pyrazol-5-amine;
3-methyl-1-(pyridin-2-yl)-4-(4-(trifluoromethyl)phenyl)-1/-/-pyrazol-5-ol;
3-methyl-4-phenyl-1 -(pyridin-2-yl)-1 /-/-pyrazol-5-amine;
2-(3,5-dimethyl-4-phenyi-1H-pyrazol-1-yl)pyridine;
3-methyl-4-phenyl-1 -(pyridin-2-yl)-1 /-/-pyrazol-5-yl trifluoromethanesulfonate;
3-methyl-1-(pyridin-2-yl)-4-(3-(trifluoromethyl)phenyl)-1 /-/-pyrazol-5-ol;
4-(5-hydroxy-3-methyl-1-(pyridin-2-yl)-1/-/-pyrazol-4-yl)benzoic acid;
4-(5-hydroxy-3-methyl-1-(pyridin-2-yl)-1 /-/-pyrazol-4-yl)-Λ/-(6-hydroxyhexyl)benzamide;
3-methyl-4-(4-(methylsulfonyl)phenyl)-1-(pyridin-2-yl)-1/-/-pyrazol-5-ol;
4-(4-(hydroxymethyl)phenyl)-3-methyl-1-(pyridin-2-yl)-1/-/-pyrazol-5-ol;
3-methyl-4-phenyl-1 -(pyrimidin-2-yl)-1 H-pyrazol-5-ol;
6-(5-hydroxy-3-methyl-4-phenylpyrazol-1-yl)-pyridazin-3-ol;
3-methyl-4-phenyl-1 -(thiazoi-2-yl)-1 /-/-pyrazol-5-ol;
1 -(benzo[c/|thiazol-2-yl)-3-methyl-4-phenyl-1 H-pyrazol-5-ol;
1 -(1 H-benzo[d]imidazol-2-yl)-3-methyl-4-phenyl-1 /-/-pyrazol-5-ol;
1 -(isoquinolin-1 -yl)-3-methyl-4-phenyl-1 H-pyrazol-5-ol;
1-(6-methoxybenzo[d]thiazol-2-yl)-3-methyl-4-phenyl-1 H-pyrazol-5-oI;
1-(6-fluorobenzo[d]thiazol-2-yl)-3-methyl-4-phenyl-1 H-pyrazol-5-ol;
3-methyl-1 -(5-nitrothiazol-2-yl)-4-phenyl-1 H-pyrazol-5-ol;
3-methyl-4-(4-nitrophenyi)-1 -(pyridin-2-yl)-1 H-pyrazol-5-ol;
4-(4-aminophenyl)-3-methyl-1-(pyridin-2-yl)-1 H-pyrazol-5-ol;
4-(4-methoxyphenyl)-3-methyl-1-(pyridin-2-yl)-1 H-pyrazol-5-ol;
4-(4-(diethylamino)phenyl)-3-methyl-1-(pyridin-2-yl)-1 H-pyrazol-5-ol;
4-(5-hydroxy-3-methyl-1-(pyridin-2-yl)-1H-pyrazol-4-yl)benzonitrile;
4-(4-fluorophenyl)-3-methyl-1 -(pyridin-2-yl)-1 H-pyrazol-5-ol; ethyl 4-(5-hydroxy-3-methyl-1-(pyridin-2-yl)-1H-pyrazol-4-yl)benzoate;
3-methyl-4-phenyl-1 -(pyridin-2-yl-methyl)-1 H-pyrazol-5-ol;
1 ,3-phenylenebis(methylene) dicarbamimidothioate;
(2-fluoro-1 ,3-phenylene)bis(methylene) dicarbamimidothioate;
1 ,3-phenylene dicarbamimidothioate;
(5-methyl-1 ,3-phenylene)bis(methylene) dicarbamimidothioate;
(2,4,6-trimethylbenzene-1 ,3,5-triyl)tris(methylene) tricarbamimidothioate;
2-{1-[3-(1-carbamimidoylsulfanyl-1-methylethyl)phenyl]-1-methylethyl}isothiourea;
(2-cyano-1 ,3-phenylene)bis(methylene) dicarbamimidothioate; (4,6-dimethyl-1 ,3-phenylene)bis(methylene) dicarbamimidothioate; diethyl 4,6-bis(carbamimidoylthiomethy!)isophthalate; (5-bromo-4,6-dimethyl-1 ,3-phenylene)bis(methylene) dicarbamimidothioate; (2,4,5,6-tetramethyl-1 ,3-phenylene)bis(methylene) dicarbamimidothioate; 2-{1-[3-(1-carbamimidoylsulfanylethyl)-2,4,6-trimethylphenyl]ethyl}isothiourea; (2-hydroxy-5-methyl-1 ,3-phenylene)bis(methylene) dicarbamimidothioate; 1 ,3-di[(methylamidino)thiomethyl]-2,4,6-trimethylbenzene; (5-hydroxy-2,4,6-trimethyl-1 ,3-phenylene)bis(methylene) dicarbamimidothioate; (2,4,5,6-tetrachloro-1 ,3-phenylene)bis(methylene) dicarbamimidothioate; (2-methoxy-5-methyl-1 ,3-phenylene)bis(methylene) dicarbamimidothioate; (2-methyl-1 ,3-phenylene)bis(methylene) dicarbamimidothioate; (4-methoxy-1 ,3-phenylene)bis(methylene) dicarbamimidothioate; (5-methoxy-1 ,3-phenylene)bis(methylene) dicarbamimidothioate; (4,6-dibromo-1 ,3-phenylene)bis(methylene) dicarbamimidothioate; (4,6-diisopropyl-1 ,3-phenylene)bis(methylene) dicarbamimidothioate; 1 ,3-di[(2-cyano-3-methylguanidino)methyl]-2,4,6-trimethyl benzene; 2,2'-(1 ,3-phenylene)diacetimidamide; Λ/-(3-guanidinomethyl-2,4,6-trimethylbenzyl)guanidine; pyridine-2,6-diylbis(methylene) dicarbamimidothioate; (2, 4,6-trimethylpyridine-3,5-diyl)bis(methylene) dicarbamimidothioate; (1 ,2-phenylene)bis(methylene) dicarbamimidothioate; (3,4,5, 6-tetramethyl-1 ,2-phenylene)bis(methylene) dicarbamimidothioate; naphthalene-1 ,2-diylbis(methylene) dicarbamimidothioate; naphthalene-1 ,8-diylbis(methylene) dicarbamimidothioate; 2-(5-carbamimidoylsulfanecarbonyl-3,4-dichlorothiophene-2-carbonyl)isothiourea; thiophene-2,5-diylbis(methylene) dicarbamimidothioate; (3,4-diphenylthiophene-2,5-diyl)bis(methyIene) dicarbamimidothioate; (3, 4-dimethylthiophene-2,5-diyl)bis(methylene) dicarbamimidothioate; (3,4-dimethylthieno[2,3-jfc)]thiophene-2,5-diyl)bis(methylene) dicarbamimidothioate; (4-amino-4H-1 ,2,4-triazole-3,5-diyl)bis(methylene) dicarbamimidothioate; or (1 /-/-1 ,2,4-triazo!e-3,5-diyl)bis(methylene) dicarbamimidothiodate; and pharmaceutically acceptable salts thereof.
8. The method of Claim 7 wherein the DMT1 inhibitor is (2,4,6-trimethyl- 1 ,3-phenylene)bis(methylene) dicarbamimidothioate dihydrochloride.
9. The method of any one of Claims 1 to 8 wherein the therapeutically effective amount of the DMT1 inhibitor is a daily dosage amount from about 0.001 mg/Kg body weight of the mammal to about 100 mg/Kg body weight of the mammal.
10. The method of Claim 9 wherein the daily dosage amount of the DMT1 inhibitor is selected from 10 mg/Kg body weight of the mammal, 25 mg/Kg body weight of the mammal, or 50 mg/Kg body weight of the mammal.
11. The method of any one of Claims 3 to 8 wherein the therapeutically effective amount of the proton pump inhibitor is a daily dosage amount from about 10 mg/Kg body weight of the mammal to about 100 mg/Kg body weight of the mammal.
12. The method of Claim 1 1 wherein the daily dosage amount of the proton pump inhibitor is selected from 15 mg/Kg body weight of the mammal, 40 mg/Kg body weight of the mammal, or 75 mg/Kg body weight of the mammal.
13. The method of any one of Claims 4 to 8 wherein the therapeutically effective amount of the DMT1 inhibitor is a daily dosage amount from about 0.001 mg/Kg body weight of the mammal to about 100 mg/Kg body weight of the mammal and the therapeutically effective amount of the proton pump inhibitor is a daily dosage amount from about 10 mg/Kg body weight of the mammal to about 100 mg/Kg body weight of the mammal.
14. The method of Claim 13 wherein the daily dosage amount of the DMT1 inhibitor is selected from 10 mg/Kg body weight of the mammal, 25 mg/Kg body weight of the mammal, or 50 mg/Kg body weight of the mammal, and the daily dosage amount of the proton pump inhibitor is selected from 15 mg/Kg body weight of the mammal, 40 mg/Kg body weight of the mammal, or 75 mg/Kg body weight of the mammal.
15. The method of Claim 14 wherein the daily dosage amount of the DMT1 inhibitor is 10 mg/Kg body weight of the mammal and the daily dosage amount of the proton pump inhibitor is 15 mg/Kg body weight of the mammal.
16. The method of Claim 14 wherein the daily dosage amount of the DMT1 inhibitor is 25 mg/Kg body weight of the mammal and the daily dosage amount of the proton pump inhibitor is 40 mg/Kg body weight of the mammal.
17. The method of Claim 14 wherein the daily dosage amount of the DMT1 inhibitor is 50 mg/Kg body weight of the mammal and the daily dosage amount of the proton pump inhibitor is 75 mg/Kg body weight of the mammal.
18. The method of any one of Claims 3 to 17 wherein the DMT1 inhibitor and the proton pump inhibitor are concomitantly administered to the mammal.
19. The method of any one of Claims 3 to 17 wherein the DMT1 inhibitor and the proton pump inhibitor are sequentially administered to the mammal.
20. The method of any one of Claims 1 to 19 wherein the iron overload disorder is a primary iron overload disorder.
21. The method of Claim 20 wherein the primary iron overload disorder is independently selected from hereditary hemochromatosis, juvenile hemochromatosis, ferroportin disease, neonatal hemochromatosis, Bantu siderosis, African iron overload, gracile syndrome, ataxia, or Friedreich Ataxia.
22. The method of any one of Claims 1 to 19 wherein the iron overload disorder is a secondary iron overload disorder.
23. The method of any one of Claims 1 to 19 wherein the iron overload disorder is transfusional iron overload disorder.
24. The method of any one of Claims 1 to 19 wherein the iron overload disorder is associated with a disease and/or condition independently selected from thalassemia (beta and alpha, major, minor and intermedia), hypochromic microcytic anemia, sickle cell anemia, microcytic iron loading anemia, hereditary sideroblastic anemia, congenital dyserythropoeitic anemia, porphyria cutanea tarda, pyruvate kinase deficiency, hereditary atransferrinemia, ceruloplasmin deficiency, myelodysplastic syndromes, pulmonary hemosiderosis, aceruloplasminemia or x-linked sideroblastic anemia.
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Cited By (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2016112381A1 (en) 2015-01-09 2016-07-14 The Board Of Trustees Of The University Of Illinois Restoring physiology in iron-deficient organisms using small molecules
CN107119009A (en) * 2017-06-30 2017-09-01 云南农业大学 A kind of porcine kidney cell iron deficiency model and its method for building up
US10780055B2 (en) 2017-10-25 2020-09-22 Chiesi Farmaceutici S.P.A. Delayed release deferiprone tablets and methods of using the same
EP3785696A1 (en) * 2019-08-28 2021-03-03 B.R.A.I.N. Biotechnology Research And Information Network AG Cosmetic use of aromatic formamidine derivatives
CN115515948A (en) * 2020-03-20 2022-12-23 阿克比治疗有限公司 PHD inhibitor compounds, compositions and uses
CN115605467A (en) * 2020-03-20 2023-01-13 阿克比治疗有限公司（Us） PHD inhibitor compounds, compositions and uses thereof
WO2023018989A1 (en) * 2021-08-13 2023-02-16 The University Of Kansas Cross-linked polymeric chelators compositions and use thereof
US12016850B2 (en) 2022-04-11 2024-06-25 Chiesi Farmaceutici S.P.A. Modified release pharmaceutical formulations comprising deferiprone
US12016851B2 (en) 2022-04-11 2024-06-25 Chiesi Farmaceutici S.P.A. Modified release pharmaceutical formulations comprising deferiprone

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1998044954A1 (en) * 1997-04-04 1998-10-15 Brigham And Women's Hospital Metal ion transporter and uses therefor
WO2008115999A1 (en) * 2007-03-19 2008-09-25 Xenon Pharmaceuticals Inc. Biaryl and biheteroaryl compounds useful in treating iron disorders
WO2008121861A2 (en) * 2007-03-28 2008-10-09 Xenon Pharmaceuticals Inc. Pyrazole and pyrrole compounds useful in treating iron disorders
WO2008151288A2 (en) * 2007-06-05 2008-12-11 Xenon Pharmaceuticals Inc. Aromatic and heteroaromatic compounds useful in treating iron disorders

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1998044954A1 (en) * 1997-04-04 1998-10-15 Brigham And Women's Hospital Metal ion transporter and uses therefor
WO2008115999A1 (en) * 2007-03-19 2008-09-25 Xenon Pharmaceuticals Inc. Biaryl and biheteroaryl compounds useful in treating iron disorders
WO2008121861A2 (en) * 2007-03-28 2008-10-09 Xenon Pharmaceuticals Inc. Pyrazole and pyrrole compounds useful in treating iron disorders
WO2008151288A2 (en) * 2007-06-05 2008-12-11 Xenon Pharmaceuticals Inc. Aromatic and heteroaromatic compounds useful in treating iron disorders

Non-Patent Citations (5)

* Cited by examiner, † Cited by third party
Title
BEUTLER ET AL: "Iron storage disease: Facts, fiction and progress", BLOOD CELLS, MOLECULES AND DISEASES, LAJOLLA, US, vol. 39, no. 2, 31 July 2007 (2007-07-31), pages 140 - 147, XP022182369, ISSN: 1079-9796 *
CHAN ET AL: "Aripiprazole treatment for psychosis associated with Friedreich's ataxia", GENERAL HOSPITAL PSYCHIATRY, ELSEVIER SCIENCE PUBLISHING, NEW YORK, NY, US, vol. 27, no. 5, 1 September 2005 (2005-09-01), pages 372, XP005077433, ISSN: 0163-8343 *
LAERIE W. POWELL: "Hereditary hemochromatosis and iron overload diseases", JOURNAL OF GASTROENTEROLOGY AND HEPATOLOGY, vol. 17, no. Suppl, 2002, pages S191 - S195, XP002551754 *
S. LUDWICZEK ET AL: "Ca2+ channel blockers reverse iron overload by a new mechanism via divalent metal transporter-1", NATURE MEDICINE, vol. 13, no. 4, April 2007 (2007-04-01), pages 448 - 454, XP002551753 *
WETLI H A ET AL: "Small-Molecule Screening Identifies the Selanazal Drug Ebselen as a Potent Inhibitor of DMT1-Mediated Iron Uptake", CHEMISTRY AND BIOLOGY, CURRENT BIOLOGY, LONDON, GB, vol. 13, no. 9, 1 September 2006 (2006-09-01), pages 965 - 972, XP025131729, ISSN: 1074-5521, [retrieved on 20060901] *

Cited By (18)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
AU2016205030B2 (en) * 2015-01-09 2021-04-01 The Board Of Trustees Of The University Of Illinois Restoring physiology in iron-deficient organisms using small molecules
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EP3242723B1 (en) * 2015-01-09 2021-10-06 The Board of Trustees of the University of Illinois Use of hinokitiol for restoring physiology in iron-deficient organisms
CN107119009A (en) * 2017-06-30 2017-09-01 云南农业大学 A kind of porcine kidney cell iron deficiency model and its method for building up
US11357731B2 (en) 2017-10-25 2022-06-14 Chiesi Farmaceutici S.P.A. Delayed release deferiprone tablets and methods of using the same
US10940115B2 (en) 2017-10-25 2021-03-09 Chiesi Farmaceutici S.P.A. Delayed release deferiprone tablets and methods of using the same
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US11458103B2 (en) 2017-10-25 2022-10-04 Chiesi Farmaceutici S.P.A. Delayed release deferiprone tablets and methods of using the same
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