Patent Application
20090291955
The present invention relates to azacyclohexane derivatives which are inhibitors of stearoyl-coenzyme A delta-9 desaturase (SCD) and the use of such compounds to control, prevent and/or treat conditions or diseases mediated by SCD activity. The compounds of the present invention are useful for the control, prevention and treatment of conditions and diseases related to abnormal lipid synthesis and metabolism, including cardiovascular disease, such as atherosclerosis; obesity; diabetes; neurological disease; metabolic syndrome; insulin resistance; cancer; and hepatic steatosis.
At least three classes of fatty acyl-coenzyme A (CoA) desaturases (delta-5, delta-6 and delta-9 desaturases) are responsible for the formation of double bonds in mono- and polyunsaturated fatty acyl-CoAs derived from either dietary sources or de novo synthesis in mammals. The delta-9 specific stearoyl-CoA desaturases (SCDs) catalyze the rate-limiting formation of the cis-double bond at the C9-C10 position in monounsaturated fatty acyl-CoAs. The preferred substrates are stearoyl-CoA and palmitoyl-CoA, with the resulting oleoyl and palmitoleoyl-CoA as the main components in the biosynthesis of phospholipids, triglycerides, cholesterol esters and wax esters (Dobrzyn and Natami, Obesity Reviews, 6: 169-174 (2005)).
The rat liver microsomal SCD protein was first isolated and characterized in 1974 (Strittmatter et al., PNAS 71: 45654569 (1974)). A number of mammalian SCD genes have since been cloned and studied from various species. For example, two genes have been identified from rat (SCD1 and SCD2, Thiede et al., J. Biol. Chem., 261, 13230-13235 (1986)), Mihara, K., J. Biochem. (Tokyo), 108: 1022-1029 (1990)); four genes from mouse (SCD1, SCD2, SCD3 and SCD4) (Miyazaki et al., J. Biol. Chem., 278: 33904-33911 (2003)); and two genes from human (SCD1 and ACOD4 (SCD2)), (Zhang, et al., Biochem. J., 340: 255-264 (1991); Beiraghi, et al., Gene, 309: 11-21 (2003); Zhang et al., Biochem. J., 388: 135-142 (2005)). The involvement of SCDs in fatty acid metabolism has been known in rats and mice since the 1970's (Oshino, N., Arch. Biochem. Biophys., 149: 378-387 (1972)). This has been further supported by the biological studies of a) Asebia mice that carry the natural mutation in the SCD1 gene (Zheng et al., Nature Genetics, 23: 268-270 (1999)), b) SCD1-null mice from targeted gene deletion (Ntambi, et al., PNAS, 99: 11482-11486 (2002), and c) the suppression of SCD1 expression during leptin-induced weight loss (Cohen et al., Science, 297: 240-243 (2002)). The potential benefits of pharmacological inhibition of SCD activity has been demonstrated with anti-sense oligonucleotide inhibitors (ASO) in mice (Jiang, et al., J. Clin. Invest., 115: 1030-1038 (2005)). ASO inhibition of SCD activity reduced fatty acid synthesis and increased fatty acid oxidation in primary mouse hepatocytes. Treatment of mice with SCD-ASOs resulted in the prevention of diet-induced obesity, reduced body adiposity, hepatomegaly, steatosis, postprandial plasma insulin and glucose levels, reduced de novo fatty acid synthesis, decreased the expression of lipogenic genes, and increased the expression of genes promoting energy expenditure in liver and adipose tissues. Thus, SCD inhibition represents a novel therapeutic strategy in the treatment of obesity and related metabolic disorders.
There is compelling evidence to support that elevated SCD activity in humans is directly implicated in several common disease processes. For example, there is an elevated hepatic lipogenesis to triglyceride secretion in non-alcoholic fatty liver disease patients (Diraison, et al., Diabetes Metabolism, 29: 478-485 (2003)); Donnelly, et al., J. Clin. Invest., 115: 1343-1351 (2005)). The postprandial de novo lipogenesis is significantly elevated in obese subjects (Marques-Lopes, et al., American Journal of Clinical Nutrition, 73: 252-261 (2001)). There is a significant correlation between a high SCD activity and an increased cardiovascular risk profile including elevated plasma triglycerides, a high body mass index and reduced plasma HDL (Attie, et al., J. Lipid Res., 43: 1899-1907 (2002)). SCD activity plays a key role in controlling the proliferation and survival of human transformed cells (Scaglia and Igal, J. Biol. Chem., (2005)).
Other than the above mentioned anti-sense oligonucleotides, inhibitors of SCD activity include non-selective thia-fatty acid substrate analogs [B. Behrouzian and P. H. Buist, Prostaglandins, Leukotrienes, and Essential Fatty Acids, 68: 107-112 (2003)], cyclopropenoid fatty acids (Raju and Reiser, J. Biol. Chem., 242: 379-384 (1967)), certain conjugated long-chain fatty acid isomers (Park, et al., Biochim. Biophys. Acta, 1486: 285-292 (2000)), a series of pyridazine derivatives disclosed in published international patent application publications WO 2005/011653, WO 2005/011654, WO 2005/011656, WO 2005/011656, and WO 2005/011657, all assigned to Xenon Pharmaceuticals, Inc., and a series of heterocyclic derivatives disclosed international patent application publications WO 2006/014168, WO 2006/034279, WO 2006/034312, WO 2006/034315, WO 2006/034338, WO 2006/034341, WO 2006/034440, WO 2006/034441, and WO 2006/034446, all assigned to Xenon Pharmaceuticals, Inc.
The present invention is concerned with novel azacyclohexane derivatives as inhibitors of stearoyl-CoA delta-9 desaturase which are useful in the treatment and/or prevention of various conditions and diseases mediated by SCD activity including those related, but not limited, to elevated lipid levels, as exemplified in non-alcoholic fatty liver disease, cardiovascular disease, obesity, diabetes, metabolic syndrome, and insulin resistance.
The role of stearoyl-coenzyme A desaturase in lipid metabolism has been described by M. Miyazaki and J. M. Ntambi, Prostaglandins, Leukotrienes, and Essential Fatty Acids, 68: 113-121 (2003). The therapeutic potential of the pharmacological manipulation of SCD activity has been described by A. Dobryzn and J. M. Ntambi, in “Stearoyl-CoA desaturase as a new drug target for obesity treatment,” Obesity Reviews, 6: 169-174 (2005).
The present invention relates to azacyclohexane derivatives of structural formula I:
These azacyclohexane derivatives are effective as inhibitors of SCD. They are therefore useful for the treatment, control or prevention of disorders responsive to the inhibition of SCD, such as diabetes, insulin resistance, lipid disorders, obesity, atherosclerosis, and metabolic syndrome.
The present invention also relates to pharmaceutical compositions comprising the compounds of the present invention and a pharmaceutically acceptable carrier.
The present invention also relates to methods for the treatment, control, or prevention of disorders, diseases, or conditions responsive to inhibition of SCD in a subject in need thereof by administering the compounds and pharmaceutical compositions of the present invention.
The present invention also relates to methods for the treatment, control, or prevention of Type 2 diabetes, insulin resistance, obesity, lipid disorders, atherosclerosis, and metabolic syndrome by administering the compounds and pharmaceutical compositions of the present invention.
The present invention also relates to methods for the treatment, control, or prevention of obesity by administering the compounds of the present invention in combination with a therapeutically effective amount of another agent known to be useful to treat the condition.
The present invention also relates to methods for the treatment, control, or prevention of Type 2 diabetes by administering the compounds of the present invention in combination with a therapeutically effective amount of another agent known to be useful to treat the condition.
The present invention also relates to methods for the treatment, control, or prevention of atherosclerosis by administering the compounds of the present invention in combination with a therapeutically effective amount of another agent known to be useful to treat the condition.
The present invention also relates to methods for the treatment, control, or prevention of lipid disorders by administering the compounds of the present invention in combination with a therapeutically effective amount of another agent known to be useful to treat the condition.
The present invention also relates to methods for treating metabolic syndrome by administering the compounds of the present invention in combination with a therapeutically effective amount of another agent known to be useful to treat the condition.
The present invention is concerned with azacyclohexane derivatives useful as inhibitors of SCD. Compounds of the present invention are described by structural formula I:
or a pharmaceutically acceptable salt thereof; wherein
each n is independently 0, 1 or 2;
p is 0, 1, or 2;
X-Y is N—C(O), N—S(O)2, N—CR1R2, CH—O, CH—S(O)p, CH—NR13, CR17—CR1R2, or CH—C(O);
Ar is phenyl, naphthyl, or heteroaryl optionally substituted with one to five R3 substituents;
HetAr is a fused heteroaromatic ring selected from the group consisting of:
wherein Z is O, S, or N—R18;
R1 and R2 are each independently hydrogen, halogen, or C1-3 alkyl, wherein alkyl is optionally substituted with one to three substituents independently selected from fluorine and hydroxy; or R1 and R2 together with the carbon atom to which they are attached can form a spirocyclopropyl ring system;
each R3 is independently selected from the group consisting of:
C1-6 alkyl,
(CH2)n-phenyl,
(CH2)n-naphthyl,
(CH2)n-heteroaryl,
(CH2)n-heterocyclyl,
(CH2)nC3-7 cycloalkyl,
halogen,
OR4,
(CH2)nN(R4)2,
(CH2)nC≡N,
(CH2)nCO2R4,
NO2,
(CH2)nNR4SO2R4
(CH2)nSO2N(R4)2,
(CH2)nS(O)pR4,
(CH2)nNR4C(O)N(R4)2,
(CH2)nC(O)N(R4)2,
(CH2)nNR4C(O)R4,
(CH2)nNR4CO2R4,
O(CH2)nC(O)N(R4)2,
CF3,
CH2CF3,
OCF3, and
OCH2CF3;
in which phenyl, naphthyl, heteroaryl, cycloalkyl, and heterocyclyl are optionally substituted with one to three substituents independently selected from halogen, hydroxy, C1-4 alkyl, trifluoromethyl, and C1-4 alkoxy; and wherein any methylene (CH2) carbon atom in R3 is optionally substituted with one to two groups independently selected from fluorine, hydroxy, and C1-4 alkyl; or two substituents when on the same methylene (CH2) group are taken together with the carbon atom to which they are attached to form a cyclopropyl group;
each R4 is independently selected from the group consisting of
hydrogen,
C1-6 alkyl,
(CH2)n-phenyl,
(CH2)n-heteroaryl,
(CH2)n-naphthyl, and
(CH2)nC3-7 cycloalkyl;
wherein alkyl, phenyl, heteroaryl, and cycloalkyl are optionally substituted with one to three groups independently selected from halogen, C1-4 alkyl, and C1-4 alkoxy; or two R4 groups together with the atom to which they are attached form a 4- to 8-membered mono- or bicyclic ring system optionally containing an additional heteroatom selected from O, S, NH, and NC1-4 alkyl;
R5, R6, R7, R8, R9, R10, R11, and R12 are each independently hydrogen, fluorine, or C1-3 alkyl, wherein alkyl is optionally substituted with one to three substituents independently selected from fluorine and hydroxy;
each R13 is independently hydrogen or C1-6 alkyl;
R14 is independently selected from the group consisting of amino, hydroxy, mercapto, C1-4 alkoxy, C1-4 alkylthio, C1-4 alkylamino, di-(C1-4 alkyl)amino, arylamino, aryl-C1-2 alkylamino, C1-4 alkylcarbonylamino, aryl-C1-2 alkylcarbonylamino, arylcarbonylamino, C1-4 alkylaminocarbonylamino, C1-4 alkylsulfonylamino, arylsulfonylamino, aryl-C1-2 alkylsulfonylamino, C1-4 alkyloxycarbonylamino, aryloxycarbonylamino, and aryl-C1-2 alkyloxycarbonylamino;
R15 and R16 are each independently hydrogen or C1-4 alkyl optionally substituted with amino, hydroxy, C1-4 alkoxy, C1-4 alkylthio, C1-4 alkylsulfonyl, C1-4 alkylcarbonyloxy, phenyl, heteroaryl, or one to five halogens;
R17 is hydrogen, C1-3 alkyl, fluorine, or hydroxy; and
R18 is selected from the group consisting of hydrogen, C1-4 alkyl, C1-4 alkylcarbonyl, aryl-C1-2 alkylcarbonyl, arylcarbonyl, C1-4 alkylaminocarbonyl, C1-4 alkylsulfonyl, arylsulfonyl, aryl-C1-2 alkylsulfonyl, C1-4 alkyloxycarbonyl, aryloxycarbonyl, and aryl-C1-2 alkyloxycarbonyl.
In one embodiment of the compounds of the present invention, n is 0.
In a second embodiment of the compounds of the present invention, X-Y is N—C(O). In a class of this embodiment, Ar is phenyl substituted with one to three R3 substituents as defined above. In a subclass of this class, phenyl is substituted at the ortho-position with one of the R3 substituents.
In a third embodiment of the compounds of the present invention, X-Y is N—S(O)2. In a class of this embodiment, Ar is phenyl substituted with one to three R3 substituents as defined above. In a subclass of this class, phenyl is substituted at the ortho-position with one of the R3 substituents.
In a fourth embodiment of the compounds of the present invention, X-Y is CH—O. In a class of this embodiment, Ar is phenyl substituted with one to three R3 substituents as defined above. In a subclass of this class, phenyl is substituted at the ortho-position with one of the R3 substituents.
In a fifth embodiment of the compounds of the present invention, X-Y is CH—S(O)p. In a class of this embodiment, Ar is phenyl substituted with one to three R3 substituents as defined above. In a subclass of this class, phenyl is substituted at the ortho-position with one of the R3 substituents.
In a sixth embodiment of the compounds of the present invention, X-Y is N—CR1R2. In a class of this embodiment, Ar is phenyl substituted with one to three R3 substituents as defined above. In yet another class of this embodiment, R1 and R2 are hydrogen and Ar is phenyl substituted with one to three R3 substituents. In a subclass of this class, phenyl is substituted at the ortho-position with one of the R3 substituents.
In a seventh embodiment of the compounds of the present invention, X-Y is CH—NR13. In a class of this embodiment, Ar is phenyl substituted with one to three R3 substituents as defined above. In yet another class of this embodiment, R13 is hydrogen and Ar is phenyl substituted with one to three R3 substituents. In a subclass of this class, In a subclass of this class, phenyl is substituted at the ortho-position with one of the R3 substituents.
In an eighth embodiment of the compounds of the present invention, X-Y is CH—C(O). In a class of this embodiment, Ar is phenyl substituted with one to three R3 substituents as defined above. In a subclass of this class, phenyl is substituted at the ortho-position with one of the R3 substituents.
In a ninth embodiment of the compounds of the present invention, X-Y is CR17—CR1R2. In a class of this embodiment, Ar is phenyl substituted with one to three R3 substituents as defined above. In yet another class of this embodiment, R1, R2, and R17 are hydrogen and Ar is phenyl substituted with one to three R3 substituents. In a subclass of this class, phenyl is substituted at the ortho-position with one of the R3 substituents.
In a further embodiment of the compounds of the present invention, R5—R12 are hydrogen.
In yet a further embodiment of the compounds of the present invention, each R3 is independently selected from the group consisting of halogen, C1-4 alkyl, trifluoromethyl, cyano, C1-4 alkoxy, C1-4 alkylthio, and phenyl.
In yet a further embodiment, HetAr is a fused heteroaromatic ring selected from the group consisting of:
wherein Z, R13, R15, and R16 are as defined above. In a class of this embodiment, Z is S, R13 is hydrogen, and R15 and R16 are each independently hydrogen, methyl, or hydroxymethyl. In another class of this embodiment, HetAr is
wherein Z, R13, and R15 are as defined above. In a subclass of this class, Z is S, and R13 is hydrogen and R15 is hydrogen, methyl, or hydroxymethyl.
Illustrative, but nonlimiting examples, of compounds of the present invention that are useful as inhibitors of SCD are the following:
and pharmaceutically acceptable salts thereof.
As used herein the following definitions are applicable.
“Alkyl”, as well as other groups having the prefix “alk”, such as alkoxy and alkanoyl, means carbon chains which may be linear or branched, and combinations thereof, unless the carbon chain is defined otherwise. Examples of alkyl groups include methyl, ethyl, propyl, isopropyl, butyl, sec- and tert-butyl, pentyl, hexyl, heptyl, octyl, nonyl, and the like. Where the specified number of carbon atoms permits, e.g., from C3-10, the term alkyl also includes cycloalkyl groups, and combinations of linear or branched alkyl chains combined with cycloalkyl structures. When no number of carbon atoms is specified, C1-6 is intended.
“Cycloalkyl” is a subset of alkyl and means a saturated carbocyclic ring having a specified number of carbon atoms. Examples of cycloalkyl include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, and the like. A cycloalkyl group generally is monocyclic unless stated otherwise. Cycloalkyl groups are saturated unless otherwise defined.
The term “alkoxy” refers to straight or branched chain alkoxides of the number of carbon atoms specified (e.g., C1-6 alkoxy), or any number within this range [i.e., methoxy (MeO—), ethoxy, isopropoxy, etc.].
The term “alkylthio” refers to straight or branched chain alkylsulfides of the number of carbon atoms specified (e.g., C1-6 alkylthio), or any number within this range [i.e., methylthio (MeS—), ethylthio, isopropylthio, etc.].
The term “alkylamino” refers to straight or branched alkylamines of the number of carbon atoms specified (e.g., C1-6 alkylamino), or any number within this range [i.e., methylamino, ethylamino, isopropylamino, t-butylamino, etc.].
The term “alkylsulfonyl” refers to straight or branched chain alkylsulfones of the number of carbon atoms specified (e.g., C1-6 alkylsulfonyl), or any number within this range [i.e., methylsulfonyl (MeSO2—), ethylsulfonyl, isopropylsulfonyl, etc.].
The term “alkylsulfinyl” refers to straight or branched chain alkylsulfoxides of the number of carbon atoms specified (e.g., C1-6 alkylsulfinyl), or any number within this range [i.e., methylsulfinyl (MeSO—), ethylsulfinyl, isopropylsulfinyl, etc.].
The term “alkyloxycarbonyl” refers to straight or branched chain esters of a carboxylic acid derivative of the present invention of the number of carbon atoms specified (e.g., C1-6 alkyloxycarbonyl), or any number within this range [i.e., methyloxycarbonyl (MeOCO—), ethyloxycarbonyl, or butyloxycarbonyl].
“Aryl” means a mono- or polycyclic aromatic ring system containing carbon ring atoms. The preferred aryls are monocyclic or bicyclic 6-10 membered aromatic ring systems. Phenyl and naphthyl are preferred aryls. The most preferred aryl is phenyl.
“Heterocyclyl” refer to saturated or unsaturated non-aromatic rings or ring systems containing at least one heteroatom selected from O, S and N, further including the oxidized forms of sulfur, namely SO and SO2. Examples of heterocycles include tetrahydrofuran (THF), dihydrofuran, 1,4-dioxane, morpholine, 1,4-dithiane, piperazine, piperidine, 1,3-dioxolane, imidazolidine, imidazoline, pyrroline, pyrrolidine, tetrahydropyran, dihydropyran, oxathiolane, dithiolane, 1,3-dioxane, 1,3-dithiane, oxathiane, thiomorpholine, 2-oxopiperidin-1-yl, 2-oxopyrrolidin-1-yl, and 2-oxoazetidin-1-yl, and the like.
“Heteroaryl” means an aromatic or partially aromatic heterocycle that contains at least one ring heteroatom selected from O, S and N. Heteroaryls thus includes heteroaryls fused to other kinds of rings, such as aryls, cycloalkyls and heterocycles that are not aromatic. Examples of heteroaryl groups include: pyrrolyl, isoxazolyl, isothiazolyl, pyrazolyl, pyridyl, oxazolyl, oxadiazolyl (in particular, 1,3,4-oxadiazol-2-yl and 1,2,4-oxadiazol-3-yl), thiadiazolyl, thiazolyl, imidazolyl, triazolyl, tetrazolyl, furyl, triazinyl, thienyl, pyrimidyl, benzisoxazolyl, benzoxazolyl, benzothiazolyl, benzothiadiazolyl, dihydrobenzofuranyl, indolinyl, pyridazinyl, indazolyl, isoindolyl, dihydrobenzothienyl, indolizinyl, cinnolinyl, phthalazinyl, quinazolinyl, naphthyridinyl, carbazolyl, benzodioxolyl, quinoxalinyl, purinyl, furazanyl, isobenzylfuranyl, benzimidazolyl, benzofuranyl, benzothienyl, quinolyl, indolyl, isoquinolyl, dibenzofuranyl, and the like. For heterocyclyl and heteroaryl groups, rings and ring systems containing from 3-15 atoms are included, forming 1-3 rings.
“Halogen” refers to fluorine, chlorine, bromine and iodine. Chlorine and fluorine are generally preferred. Fluorine is most preferred when the halogens are substituted on an alkyl or alkoxy group (e.g. CF3O and CF3CH2O).
Compounds of structural formula I may contain one or more asymmetric centers and can thus occur as racemates and racemic mixtures, single enantiomers, diastereomeric mixtures and individual diastereomers. The present invention is meant to comprehend all such isomeric forms of the compounds of structural formula I.
Compounds of structural formula I may be separated into their individual diastereoisomers by, for example, fractional crystallization from a suitable solvent, for example methanol or ethyl acetate or a mixture thereof, or via chiral chromatography using an optically active stationary phase. Absolute stereochemistry may be determined by X-ray crystallography of crystalline products or crystalline intermediates which are derivatized, if necessary, with a reagent containing an asymmetric center of known absolute configuration.
Alternatively, any stereoisomer of a compound of the general structural formula I may be obtained by stereospecific synthesis using optically pure starting materials or reagents of known absolute configuration.
If desired, racemic mixtures of the compounds may be separated so that the individual enantiomers are isolated. The separation can be carried out by methods well known in the art, such as the coupling of a racemic mixture of compounds to an enantiomerically pure compound to form a diastereomeric mixture, followed by separation of the individual diastereomers by standard methods, such as fractional crystallization or chromatography. The coupling reaction is often the formation of salts using an enantiomerically pure acid or base. The diasteromeric derivatives may then be converted to the pure enantiomers by cleavage of the added chiral residue. The racemic mixture of the compounds can also be separated directly by chromatographic methods utilizing chiral stationary phases, which methods are well known in the art.
Some of the compounds described herein contain olefinic double bonds, and unless specified otherwise, are meant to include both E and Z geometric isomers.
Some of the compounds described herein may exist as tautomers, which have different points of attachment of hydrogen accompanied by one or more double bond shifts. For example, a ketone and its enol form are keto-enol tautomers. The individual tautomers as well as mixtures thereof are encompassed with compounds of the present invention.
It will be understood that, as used herein, references to the compounds of structural formula I are meant to also include the pharmaceutically acceptable salts, and also salts that are not pharmaceutically acceptable when they are used as precursors to the free compounds or their pharmaceutically acceptable salts or in other synthetic manipulations.
The compounds of the present invention may be administered in the form of a pharmaceutically acceptable salt. The term “pharmaceutically acceptable salt” refers to salts prepared from pharmaceutically acceptable non-toxic bases or acids including inorganic or organic bases and inorganic or organic acids. Salts of basic compounds encompassed within the term “pharmaceutically acceptable salt” refer to non-toxic salts of the compounds of this invention which are generally prepared by reacting the free base with a suitable organic or inorganic acid. Representative salts of basic compounds of the present invention include, but are not limited to, the following: acetate, benzenesulfonate, benzoate, bicarbonate, bisulfate, bitartrate, borate, bromide, camsylate, carbonate, chloride, clavulanate, citrate, edetate, edisylate, estolate, esylate, fumarate, gluceptate, gluconate, glutamate, hexylresorcinate, hydrobromide, hydrochloride, hydroxynaphthoate, iodide, isothionate, lactate, lactobionate, laurate, malate, maleate, mandelate, mesylate, methylbromide, methylnitrate, methylsulfate, mucate, napsylate, nitrate, N-methylglucamine ammonium salt, oleate, oxalate, pamoate (embonate), palmitate, pantothenate, phosphate/diphosphate, polygalacturonate, salicylate, stearate, sulfate, subacetate, succinate, tannate, tartrate, teoclate, tosylate, triethiodide and valerate. Furthermore, where the compounds of the invention carry an acidic moiety, suitable pharmaceutically acceptable salts thereof include, but are not limited to, salts derived from inorganic bases including aluminum, ammonium, calcium, copper, ferric, ferrous, lithium, magnesium, manganic, mangamous, potassium, sodium, zinc, and the like. Particularly preferred are the ammonium, calcium, magnesium, potassium, and sodium salts. Salts derived from pharmaceutically acceptable organic non-toxic bases include salts of primary, secondary, and tertiary amines, cyclic amines, and basic ion-exchange resins, such as arginine, betaine, caffeine, choline, N,N-dibenzylethylenediamine, diethylamine, 2-diethylaminoethanol, 2-dimethylaminoethanol, ethanolamine, ethylenediamine, N-ethylmorpholine, N-ethylpiperidine, glucamine, glucosamine, histidine, isopropylamine, lysine, methylglucamine, morpholine, piperazine, piperidine, polyamine resins, procaine, purines, theobromine, triethylamine, trimethylamine, tripropylamine, tromethamine, and the like.
Also, in the case of a carboxylic acid (—COOH) or alcohol group being present in the compounds of the present invention, pharmaceutically acceptable esters of carboxylic acid derivatives, such as methyl, ethyl, or pivaloyloxymethyl, or acyl derivatives of alcohols, such as acetyl, pivaloyl, benzoyl, and aminoacyl, can be employed. Included are those esters and acyl groups known in the art for modifying the solubility or hydrolysis characteristics for use as sustained-release or prodrug formulations.
Solvates, in particular hydrates, of the compounds of structural formula I are included in the present invention as well.
The subject compounds are useful in a method of inhibiting the stearoyl-coenzyme A delta-9 desaturase enzyme (SCD) in a patient such as a mammal in need of such inhibition comprising the administration of an effective amount of the compound. The compounds of the present invention are therefore useful to control, prevent, and/or treat conditions and diseases mediated by high or abnormal SCD enzyme activity.
Thus, one aspect of the present invention concerns a method of treating hyperglycemia, diabetes or insulin resistance in a mammalian patient in need of such treatment, which comprises administering to said patient an effective amount of a compound in accordance with structural formula I or a pharmaceutically salt or solvate thereof.
A second aspect of the present invention concerns a method of treating non-insulin dependent diabetes mellitus (Type 2 diabetes) in a mammalian patient in need of such treatment comprising administering to the patient an antidiabetic effective amount of a compound in accordance with structural formula I.
A third aspect of the present invention concerns a method of treating obesity in a mammalian patient in need of such treatment comprising administering to said patient a compound in accordance with structural formula I in an amount that is effective to treat obesity.
A fourth aspect of the invention concerns a method of treating metabolic syndrome and its sequelae in a mammalian patient in need of such treatment comprising administering to said patient a compound in accordance with structural formula I in an amount that is effective to treat metabolic syndrome and its sequelae. The sequelae of the metabolic syndrome include hypertension, elevated blood glucose levels, high triglycerides, and low levels of HDL cholesterol.
A fifth aspect of the invention concerns a method of treating a lipid disorder selected from the group consisting of dyslipidemia, hyperlipidemia, hypertriglyceridemia, hypercholesterolemia, low HDL and high LDL in a mammalian patient in need of such treatment comprising administering to said patient a compound in accordance with structural formula I in an amount that is effective to treat said lipid disorder.
A sixth aspect of the invention concerns a method of treating atherosclerosis in a mammalian patient in need of such treatment comprising administering to said patient a compound in accordance with structural formula I in an amount effective to treat atherosclerosis.
A seventh aspect of the invention concerns a method of treating cancer in a mammalian patient in need of such treatment comprising administering to said patient a compound in accordance with structural formula I in an amount effective to treat cancer.
A further aspect of the invention concerns a method of treating a condition selected from the group consisting of (1) hyperglycemia, (2) low glucose tolerance, (3) insulin resistance, (4) obesity, (5) lipid disorders, (6) dyslipidemia, (7) hyperlipidemia, (8) hypertriglyceridemia, (9) hypercholesterolemia, (10) low HDL levels, (11) high LDL levels, (12) atherosclerosis and its sequelae, (13) vascular restenosis, (14) pancreatitis, (15) abdominal obesity, (16) neurodegenerative disease, (17) retinopathy, (18) nephropathy, (19) neuropathy, (20) fatty liver disease, (21) polycystic ovary syndrome, (22) sleep-disordered breathing, (23) metabolic syndrome, and (24) other conditions and disorders where insulin resistance is a component, in a mammalian patient in need of such treatment comprising administering to the patient a compound in accordance with structural formula I in an amount that is effective to treat said condition.
Yet a further aspect of the invention concerns a method of delaying the onset of a condition selected from the group consisting of (1) hyperglycemia, (2) low glucose tolerance, (3) insulin resistance, (4) obesity, (5) lipid disorders, (6) dyslipidemia, (7) hyperlipidemia, (8) hypertriglyceridemia, (9) hypercholesterolemia, (10) low HDL levels, (11) high LDL levels, (12) atherosclerosis and its sequelae, (13) vascular restenosis, (14) pancreatitis, (15) abdominal obesity, (16) neurodegenerative disease, (17) retinopathy, (18) nephropathy, (19) neuropathy, (20) fatty liver disease, (21) polycystic ovary syndrome, (22) sleep-disordered breathing, (23) metabolic syndrome, and (24) other conditions and disorders where insulin resistance is a component, and other conditions and disorders where insulin resistance is a component, in a mammalian patient in need of such treatment comprising administering to the patient a compound in accordance with structural formula I in an amount that is effective to delay the onset of said condition.
Yet a further aspect of the invention concerns a method of reducing the risk of developing a condition selected from the group consisting of (1) hyperglycemia, (2) low glucose tolerance, (3) insulin resistance, (4) obesity, (5) lipid disorders, (6) dyslipidemia, (7) hyperlipidemia, (8) hypertriglyceridemia, (9) hypercholesterolemia, (10) low HDL levels, (11) high LDL levels, (12) atherosclerosis and its sequelae, (13) vascular restenosis, (14) pancreatitis, (15) abdominal obesity, (16) neurodegenerative disease, (17) retinopathy, (18) nephropathy, (19) neuropathy, (20) fatty liver disease, (21) polycystic ovary syndrome, (22) sleep-disordered breathing, (23) metabolic syndrome, and (24) other conditions and disorders where insulin resistance is a component, in a mammalian patient in need of such treatment comprising administering to the patient a compound in accordance with structural formula I in an amount that is effective to reduce the risk of developing said condition.
In addition to primates, such as humans, a variety of other mammals can be treated according to the method of the present invention. For instance, mammals including, but not limited to, cows, sheep, goats, horses, dogs, cats, guinea pigs, rats or other bovine, ovine, equine, canine, feline, rodent, such as a mouse, species can be treated. However, the method can also be practiced in other species, such as avian species (e.g., chickens).
The present invention is further directed to a method for the manufacture of a medicament for inhibiting stearoyl-coenzyme A delta-9 desaturase enzyme activity in humans and animals comprising combining a compound of the present invention with a pharmaceutically acceptable carrier or diluent. More particularly, the present invention is directed to the use of a compound of structural formula I in the manufacture of a medicament for use in treating a condition selected from the group consisting of hyperglycemia, Type 2 diabetes, insulin resistance, obesity, and a lipid disorder in a mammal, wherein the lipid disorder is selected from the group consisting of dyslipidemia, hyperlipidemia, hypertriglyceridemia, hypercholesterolemia, low HDL, and high LDL.
The subject treated in the present methods is generally a mammal, preferably a human being, male or female, in whom inhibition of stearoyl-coenzyme A delta-9 desaturase enzyme activity is desired. The term “therapeutically effective amount” means the amount of the subject compound that will elicit the biological or medical response of a tissue, system, animal or human that is being sought by the researcher, veterinarian, medical doctor or other clinician.
The term “composition” as used herein is intended to encompass a product comprising the specified ingredients in the specified amounts, as well as any product which results, directly or indirectly, from combination of the specified ingredients in the specified amounts. Such term in relation to pharmaceutical composition, is intended to encompass a product comprising the active ingredient(s) and the inert ingredient(s) that make up the carrier, as well as any product which results, directly or indirectly, from combination, complexation or aggregation of any two or more of the ingredients, or from dissociation of one or more of the ingredients, or from other types of reactions or interactions of one or more of the ingredients. Accordingly, the pharmaceutical compositions of the present invention encompass any composition made by admixing a compound of the present invention and a pharmaceutically acceptable carrier. By “pharmaceutically acceptable” it is meant the carrier, diluent or excipient must be compatible with the other ingredients of the formulation and not deleterious to the recipient thereof.
The terms “administration of” and or “administering a” compound should be understood to mean providing a compound of the invention or a prodrug of a compound of the invention to the individual in need of treatment.
The utility of the compounds in accordance with the present invention as inhibitors of stearoyl-coenzyme A delta-9 desaturase (SCD) enzyme activity may be demonstrated by the following microsomal and whole-cell based assays:
The activity of compounds of formula I against the SCD enzyme is determined by following the conversion of radiolabeled-stearoyl-CoA to oleoyl-CoA using SCD1-induced rat liver microsome and a previously published procedure with some modifications (Joshi, et al., J. Lipid Res., 18: 32-36 (1977)). After feeding wistar rats with a high carbohydrate/fat-free rodent diet (LabDiet #5803, Purina) for 3 days, the SCD-induced livers were homogenized (1:10 w/v) in 250 mM sucrose, 1 mM EDTA, 5 mM DTT and 50 mM Tris-HCl (pH 7.5). After a 20 min centrifugation (18,000×g/4° C.) to remove tissue and cell debris, the microsome was prepared by a 100,000×g centrifugation (60 min) with the resulting pellet suspended in 100 mM sodium phosphate, 20% glycerol and 2 mM DTT. Test compound in 2 μL DMSO was incubated for 15 min. at room temperature with 180 μL of the microsome (typically at about 100 μg/mL, in Tris-HCl buffer (100 mM, pH 7.5), ATP (5 mM), Coenzyme A (0.1 mM), Triton X-100 (0.5 mM) and NADH (2 mM)). The reaction was initiated by the addition of 20 μL of [3H]-Stearoyl-CoA (final concentration at 2 μM with the radioactivity concentration at 1 μCi/mL), and terminated by the addition of 150 μL of 1N sodium hydroxide. After 60 min at room temperature to hydrolyze the oleoyl-CoA and stearoyl-CoA, the solution was acidified by the addition of 150 μL of 15% phosphoric acid (v/v) in ethanol supplemented with 0.5 mg/mL stearic acid and 0.5 mg/mL oleic acid. [3H]-oleic acid and [3H]-stearic acid were then quantified on a HPLC that is equipped with a C-18 reverse phase column and a Packard Flow Scintillation Analyzer. Alternatively, the reaction mixture (80 μL) was mixed with a calcium chloride/charcoal aqueous suspension (100 μL of 15% (w/v) charcoal plus 20 μL of 2 N CaCl2). The resulting mixture was centrifuged to precipitate the radioactive fatty acid species into a stable pellet. Tritiated water from SCD-catalyzed desaturation of 9,10-[3H]-stearoyl-CoA was quantified by counting 50 μL of the supernant on a scintillation counter.
Human HepG2 cells were grown on 24-well plates in MEM media (Gibco cat#11095-072) supplemented with 10% heat-inactivated fetal bovine serum at 37° C. under 5% CO2 in a humidified incubator. Test compound dissolved in the media was incubated with the subconfluent cells for 15 min at 37° C. [1-14C]-stearic acid was added to each well to a final concentration of 0.05 μCi/mL to detect SCD-catalyzed [14C]-oleic acid formation. 0.05 μCi/mL of [1-14C]-eicosatrienoic acid or [1-14C]-linolenic acid plus 10 μM of 2-amino-N-(3-chlorophenyl)benzamide (a delta-5 desaturase inhibitor) was used to index the delta-5 and delta-6 desaturase activities, respectively. After 4 h incubation at 37° C., the culture media was removed and the labeled cells were washed with PBS (3×1 mL) at room temperature. The labeled cellular lipids were hydrolyzed under nitrogen at 65° C. for 1 h using 400 μL of 2N sodium hydroxide plus 50 μL of L-α-phosphatidylcholine (2 mg/mL in isopropanol, Sigma #P-3556). After acidification with phosphoric acid (60 μL), the radioactive species were extracted with 300 μL of acetonitrile and quantified on a HPLC that was equipped with a C-18 reverse phase column and a Packard Flow Scintillation Analyzer. The levels of [14C]-oleic acid over [14C]-stearic acid, [14C]-arachidonic acid over [14C]-eicosatrienoic acid, and [14C]-eicosatetraenoic acid (8,11,14,17) over [14C]-linolenic acid were used as the corresponding activity indices of SCD, delta-5 and delta-6 desaturase, respectively.
The SCD inhibitors of formula I generally exhibit an inhibition constant IC50 of less than 1 μM and more typically less than 0.1 μM. Generally, the IC50 ratio for delta-5 or delta-6 desaturases to SCD for a compound of formula I is at least about ten or more, and preferably about hundred or more.
The in vivo efficacy of compounds of formula I was determined by following the conversion of [1-14C]-stearic acid to [1-14C]-oleic acid in animals as exemplified below. Mice were dosed with a compound of formula I and one hour later the radioactive tracer, [1-14C]-stearic acid, was dosed at 20 μCi/kg IV. At 3 h post dosing of the compound, the liver was harvested and then hydrolyzed in 10 N sodium hydroxide for 24 h at 80° C., to obtain the total liver fatty acid pool. After phosphoric acid acidification of the extract, the amount of [1-14C]-stearic acid and [1-14C]-oleic acid was quantified on a HPLC that was equipped with a C-18 reverse phase column and a Packard Flow Scintillation Analyzer.
The subject compounds are further useful in a method for the prevention or treatment of the aforementioned diseases, disorders and conditions in combination with other agents.
The compounds of the present invention may be used in combination with one or more other drugs in the treatment, prevention, suppression or amelioration of diseases or conditions for which compounds of Formula I or the other drugs may have utility, where the combination of the drugs together are safer or more effective than either drug alone. Such other drug(s) may be administered, by a route and in an amount commonly used therefor, contemporaneously or sequentially with a compound of Formula I. When a compound of Formula I is used contemporaneously with one or more other drugs, a pharmaceutical composition in unit dosage form containing such other drugs and the compound of Formula I is preferred. However, the combination therapy may also include therapies in which the compound of formula I and one or more other drugs are administered on different overlapping schedules. It is also contemplated that when used in combination with one or more other active ingredients, the compounds of the present invention and the other active ingredients may be used in lower doses than when each is used singly. Accordingly, the pharmaceutical compositions of the present invention include those that contain one or more other active ingredients, in addition to a compound of Formula I.
Examples of other active ingredients that may be administered in combination with a compound of formula I, and either administered separately or in the same pharmaceutical composition, include, but are not limited to:
(a) dipeptidyl peptidase IV (DPP-IV) inhibitors;
(b) insulin sensitizers including (i) PPARγ agonists, such as the glitazones (e.g. troglitazone, pioglitazone, englitazone, MCC-555, rosiglitazone, balaglitazone, and the like) and other PPAR ligands, including PPARα/γ dual agonists, such as KRP-297, muraglitazar, naveglitazar, Galida, TAK-559, PPARα agonists, such as fenofibric acid derivatives (gemfibrozil, clofibrate, fenofibrate and bezafibrate), and selective PPARγ modulators (SPPARγM's), such as disclosed in WO 02/060388, WO 02/08188, WO 2004/019869, WO 2004/020409, WO 2004/020408, and WO 2004/066963; (ii) biguanides such as metformin and phenformin, and (iii) protein tyrosine phosphatase-1B (PTP-1B) inhibitors;
(c) insulin or insulin mimetics;
(d) sulfonylureas and other insulin secretagogues, such as tolbutamide, glyburide, glipizide, glimepiride, and meglitinides, such as nateglinide and repaglinide;
(e) α-glucosidase inhibitors (such as acarbose and miglitol);
(f) glucagon receptor antagonists, such as those disclosed in WO 98/04528, WO 99/01423, WO 00/39088, and WO 00/69810;
(g) GLP-1, GLP-1 analogues or mimetics, and GLP-1 receptor agonists, such as exendin-4 (exenatide), liraglutide (N,N-2211), CJC-1131, LY-307161, and those disclosed in WO 00/42026 and WO 00/59887;
(h) GIP and GIP mimetics, such as those disclosed in WO 00/58360, and GIP receptor agonists;
(i) PACAP, PACAP mimetics, and PACAP receptor agonists such as those disclosed in WO 01/23420;
(j) cholesterol lowering agents such as (i) HMG-CoA reductase inhibitors (lovastatin, simvastatin, pravastatin, cerivastatin, fluvastatin, atorvastatin, itavastatin, and rosuvastatin, and other statins), (ii) sequestrants (cholestyramine, colestipol, and dialkylaminoalkyl derivatives of a cross-linked dextran), (iii) nicotinyl alcohol, nicotinic acid or a salt thereof, (iv) PPARα agonists such as fenofibric acid derivatives (gemfibrozil, clofibrate, fenofibrate and bezafibrate), (v) PPARα/γ dual agonists, such as naveglitazar and muraglitazar, (vi) inhibitors of cholesterol absorption, such as beta-sitosterol and ezetimibe, (vii) acyl CoA:cholesterol acyltransferase inhibitors, such as avasimibe, and (viii) antioxidants, such as probucol;
(k) PPARδ agonists, such as those disclosed in WO 97/28149;
(l) antiobesity compounds, such as fenfluramine, dexfenfluramine, phentermine, sibutramine, orlistat, neuropeptide Y1 or Y5 antagonists, CB1 receptor inverse agonists and antagonists, β3 adrenergic receptor agonists, melanocortin-receptor agonists, in particular melanocortin-4 receptor agonists, ghrelin antagonists, bombesin receptor agonists (such as bombesin receptor subtype-3 agonists), and melanin-concentrating hormone (MCH) receptor antagonists;
(m) ileal bile acid transporter inhibitors;
(n) agents intended for use in inflammatory conditions such as aspirin, non-steroidal anti-inflammatory drugs (NSAIDs), glucocorticoids, azulfidine, and selective cyclooxygenase-2 (COX-2) inhibitors;
(o) antihypertensive agents, such as ACE inhibitors (enalapril, lisinopril, captopril, quinapril, tandolapril), A-II receptor blockers (losartan, candesartan, irbesartan, valsartan, telmisartan, and eprosartan), beta blockers and calcium channel blockers;
(p) glucokinase activators (GKAs), such as those disclosed in WO 03/015774; WO 04/076420; and WO 04/081001;
(q) inhibitors of 11β-hydroxysteroid dehydrogenase type 1, such as those disclosed in U.S. Pat. No. 6,730,690; WO 03/104207; and WO 04/058741;
(r) inhibitors of cholesteryl ester transfer protein (CETP), such as torcetrapib; and
(s) inhibitors of fructose 1,6-bisphosphatase, such as those disclosed in U.S. Pat. Nos. 6,054,587; 6,110,903; 6,284,748; 6,399,782; and 6,489,476.
Dipeptidyl peptidase-IV inhibitors that can be combined with compounds of structural formula I include those disclosed in U.S. Pat. No. 6,699,871; WO 02/076450 (3 Oct. 2002); WO 03/004498 (16 Jan. 2003); WO 03/004496 (16 Jan. 2003); EP 1 258 476 (20 Nov. 2002); WO 02/083128 (24 Oct. 2002); WO 02/062764 (15 Aug. 2002); WO 03/000250 (3 Jan. 2003); WO 03/002530 (9 Jan. 2003); WO 03/002531 (9 Jan. 2003); WO 03/002553 (9 Jan. 2003); WO 03/002593 (9 Jan. 2003); WO 03/000180 (3 Jan. 2003); WO 03/082817 (9 Oct. 2003); WO 03/000181 (3 Jan. 2003); WO 04/007468 (22 Jan. 2004); WO 04/032836 (24 Apr. 2004); WO 04/037169 (6 May 2004); and WO 04/043940 (27 May 2004). Specific DPP-IV inhibitor compounds include isoleucine thiazolidide (P32/98); NVP-DPP-728; LAF 237; P93/01; and saxagliptin (BMS 477118).
Antiobesity compounds that can be combined with compounds of structural formula I include fenfluramine, dexfenfluramine, phentermine, sibutramine, orlistat, neuropeptide Y1 or Y5 antagonists, cannabinoid CB1 receptor antagonists or inverse agonists, melanocortin receptor agonists, in particular, melanocortin-4 receptor agonists, ghrelin antagonists, bombesin receptor agonists, and melanin-concentrating hormone (MCH) receptor antagonists. For a review of anti-obesity compounds that can be combined with compounds of structural formula I, see S. Chaki et al., “Recent advances in feeding suppressing agents: potential therapeutic strategy for the treatment of obesity,” Expert Opin. Ther. Patents, 11: 1677-1692 (2001); D. Spanswick and K. Lee, “Emerging antiobesity drugs,” Expert Opin. Emerging Drugs, 8: 217-237 (2003); and J. A. Fernandez-Lopez, et al., “Pharmacological Approaches for the Treatment of Obesity,” Drugs, 62: 915-944 (2002).
Neuropeptide Y5 antagonists that can be combined with compounds of structural formula I include those disclosed in U.S. Pat. No. 6,335,345 (1 Jan. 2002) and WO 01/14376 (1 Mar. 2001); and specific compounds identified as GW 59884A; GW 569180A; LY366377; and CGP-71683A.
Cannabinoid CB1 receptor antagonists that can be combined with compounds of formula I include those disclosed in PCT Publication WO 03/007887; U.S. Pat. No. 5,624,941, such as rimonabant; PCT Publication WO 02/076949, such as SLV-319; U.S. Pat. No. 6,028,084; PCT Publication WO 98/41519; PCT Publication WO 00/10968; PCT Publication WO 99/02499; U.S. Pat. No. 5,532,237; U.S. Pat. No. 5,292,736; PCT Publication WO 03/086288; PCT Publication WO 03/087037; PCT Publication WO 04/048317; PCT Publication WO 03/007887; PCT Publication WO 03/063781; PCT Publication WO 03/075660; PCT Publication WO 03/077847; PCT Publication WO 03/082190; PCT Publication WO 03/082191; PCT Publication WO 03/087037; PCT Publication WO 03/086288; PCT Publication WO 04/012671; PCT Publication WO 04/029204; PCT Publication WO 04/040040; PCT Publication WO 01/64632; PCT Publication WO 01/64633; and PCT Publication WO 01/64634.
Melanocortin-4 receptor (MC4R) agonists useful in the present invention include, but are not limited to, those disclosed in U.S. Pat. No. 6,294,534, U.S. Pat. Nos. 6,350,760, 6,376,509, 6,410,548, 6,458,790, U.S. Pat. No. 6,472,398, U.S. Pat. No. 5,837,521, U.S. Pat. No. 6,699,873, which are hereby incorporated by reference in their entirety; in US Patent Application Publication Nos. US 2002/0004512, US2002/0019523, US2002/0137664, US2003/0236262, US2003/0225060, US2003/0092732, US2003/109556, US 2002/0177151, US 2002/187932, US 2003/0113263, which are hereby incorporated by reference in their entirety; and in WO 99/64002, WO 00/74679, WO 02/15909, WO 01/70708, WO 01/70337, WO 01/91752, WO 02/068387, WO 02/068388, WO 02/067869, WO 03/007949, WO 2004/024720, WO 2004/089307, WO 2004/078716, WO 2004/078717, WO 2004/037797, WO 01/58891, WO 02/070511, WO 02/079146, WO 03/009847, WO 03/057671, WO 03/068738, WO 03/092690, WO 02/059095, WO 02/059107, WO 02/059108, WO 02/059117, WO 02/085925, WO 03/004480, WO 03/009850, WO 03/013571, WO 03/031410, WO 03/053927, WO 03/061660, WO 03/066597, WO 03/094918, WO 03/099818, WO 04/037797, WO 04/048345, WO 02/018327, WO 02/080896, WO 02/081443, WO 03/066587, WO 03/066597, WO 03/099818, WO 02/062766, WO 03/000663, WO 03/000666, WO 03/003977, WO 03/040107, WO 03/040117, WO 03/040118, WO 03/013509, WO 03/057671, WO 02/079753, WO 02//092566, WO 03/-093234, WO 03/095474, and WO 03/104761.
One particular aspect of combination therapy concerns a method of treating a condition selected from the group consisting of hypercholesterolemia, atherosclerosis, low HDL levels, high LDL levels, hyperlipidemia, hypertriglyceridemia, and dyslipidemia, in a mammalian patient in need of such treatment comprising administering to the patient a therapeutically effective amount of a compound of structural formula I and an HMG-CoA reductase inhibitor.
More particularly, this aspect of combination therapy concerns a method of treating a condition selected from the group consisting of hypercholesterolemia, atherosclerosis, low HDL levels, high LDL levels, hyperlipidemia, hypertriglyceridemia and dyslipidemia in a mammalian patient in need of such treatment wherein the HMG-CoA reductase inhibitor is a statin selected from the group consisting of lovastatin, simvastatin, pravastatin, cerivastatin, fluvastatin, atorvastatin, and rosuvastatin.
In another aspect of the invention, a method of reducing the risk of developing a condition selected from the group consisting of hypercholesterolemia, atherosclerosis, low HDL levels, high LDL levels, hyperlipidemia, hypertriglyceridemia and dyslipidemia, and the sequelae of such conditions is disclosed comprising administering to a mammalian patient in need of such treatment a therapeutically effective amount of a compound of structural formula I and an HMG-CoA reductase inhibitor.
In another aspect of the invention, a method for delaying the onset or reducing the risk of developing atherosclerosis in a human patient in need of such treatment is disclosed comprising administering to said patient an effective amount of a compound of structural formula I and an HMG-CoA reductase inhibitor.
More particularly, a method for delaying the onset or reducing the risk of developing atherosclerosis in a human patient in need of such treatment is disclosed, wherein the HMG-CoA reductase inhibitor is a statin selected from the group consisting of: lovastatin, simvastatin, pravastatin, cerivastatin, fluvastatin, atorvastatin, and rosuvastatin.
In another aspect of the invention, a method for delaying the onset or reducing the risk of developing atherosclerosis in a human patient in need of such treatment is disclosed, wherein the HMG-Co A reductase inhibitor is a statin and further comprising administering a cholesterol absorption inhibitor.
More particularly, in another aspect of the invention, a method for delaying the onset or reducing the risk of developing atherosclerosis in a human patient in need of such treatment is disclosed, wherein the HMG-Co A reductase inhibitor is a statin and the cholesterol absorption inhibitor is ezetimibe.
In another aspect of the invention, a pharmaceutical composition is disclosed which comprises:
(1) a compound of structural formula I;
(2) a compound selected from the group consisting of:
(a) dipeptidyl peptidase IV (DPP-IV) inhibitors;
(b) insulin sensitizers including (i) PPARγ agonists, such as the glitazones (e.g. troglitazone, pioglitazone, englitazone, MCC-555, rosiglitazone, balaglitazone, and the like) and other PPAR ligands, including PPARα/γ dual agonists, such as KRP-297, muraglitazar, naveglitazar, Galida, TAK-559, PPARα agonists, such as fenofibric acid derivatives (gemfibrozil, clofibrate, fenofibrate and bezafibrate), and selective PPARγ modulators (SPPARγM's), such as disclosed in WO 02/060388, WO 02/08188, WO 2004/019869, WO 2004/020409, WO 2004/020408, and WO 2004/066963; (ii) biguanides such as metformin and phenformin, and (iii) protein tyrosine phosphatase-1B (PTP-1B) inhibitors;
(c) insulin or insulin mimetics;
(d) sulfonylureas and other insulin secretagogues, such as tolbutamide, glyburide, glipizide, glimepiride, and meglitinides, such as nateglinide and repaglinide;
(e) α-glucosidase inhibitors (such as acarbose and miglitol);
(f) glucagon receptor antagonists, such as those disclosed in WO 98/04528, WO 99/01423, WO 00/39088, and WO 00/69810;
(g) GLP-1, GLP-1 analogues or mimetics, and GLP-1 receptor agonists, such as exendin-4 (exenatide), liraglutide (N,N-2211), CJC-1131, LY-307161, and those disclosed in WO 00/42026 and WO 00/59887;
(h) GIP and GIP mimetics, such as those disclosed in WO 00/58360, and GIP receptor agonists;
(i) PACAP, PACAP mimetics, and PACAP receptor agonists such as those disclosed in WO 01/23420;
(j) cholesterol lowering agents such as (i) HMG-CoA reductase inhibitors (lovastatin, simvastatin, pravastatin, cerivastatin, fluvastatin, atorvastatin, itavastatin, and rosuvastatin, and other statins), (ii) sequestrants (cholestyramine, colestipol, and dialkylaminoalkyl derivatives of a cross-linked dextran), (iii) nicotinyl alcohol, nicotinic acid or a salt thereof, (iv) PPARα agonists such as fenofibric acid derivatives (gemfibrozil, clofibrate, fenofibrate and bezafibrate), (v) PPARα/γ dual agonists, such as naveglitazar and muraglitazar, (vi) inhibitors of cholesterol absorption, such as beta-sitosterol and ezetimibe, (vii) acyl CoA:cholesterol acyltransferase inhibitors, such as avasimibe, and (viii) antioxidants, such as probucol;
(k) PPARδ agonists, such as those disclosed in WO 97/28149;
(l) antiobesity compounds, such as fenfluramine, dexfenfluramine, phentermine, sibutramine, orlistat, neuropeptide Y1 or Y5 antagonists, CB1 receptor inverse agonists and antagonists, β3 adrenergic receptor agonists, melanocortin-receptor agonists, in particular melanocortin-4 receptor agonists, ghrelin antagonists, bombesin receptor agonists (such as bombesin receptor subtype-3 agonists), and melanin-concentrating hormone (MCH) receptor antagonists;
(m) ileal bile acid transporter inhibitors;
(n) agents intended for use in inflammatory conditions such as aspirin, non-steroidal anti-inflammatory drugs (NSAIDs), glucocorticoids, azulfidine, and selective cyclooxygenase-2 (COX-2) inhibitors;
(o) antihypertensive agents, such as ACE inhibitors (enalapril, lisinopril, captopril, quinapril, tandolapril), A-II receptor blockers (losartan, candesartan, irbesartan, valsartan, telmisartan, and eprosartan), beta blockers and calcium channel blockers;
(p) glucokinase activators (GKAs), such as those disclosed in WO 03/015774; WO 04/076420; and WO 04/081001;
(q) inhibitors of 11β-hydroxysteroid dehydrogenase type 1, such as those disclosed in U.S. Pat. No. 6,730,690; WO 03/104207; and WO 04/058741;
(r) inhibitors of cholesteryl ester transfer protein (CETP), such as torcetrapib; and
(s) inhibitors of fructose 1,6-bisphosphatase, such as those disclosed in U.S. Pat. Nos. 6,054,587; 6,110,903; 6,284,748; 6,399,782; and 6,489,476; and
(3) a pharmaceutically acceptable carrier.
When a compound of the present invention is used contemporaneously with one or more other drugs, a pharmaceutical composition containing such other drugs in addition to the compound of the present invention is preferred. Accordingly, the pharmaceutical compositions of the present invention include those that also contain one or more other active ingredients, in addition to a compound of the present invention.
The weight ratio of the compound of the present invention to the second active ingredient may be varied and will depend upon the effective dose of each ingredient. Generally, an effective dose of each will be used. Thus, for example, when a compound of the present invention is combined with another agent, the weight ratio of the compound of the present invention to the other agent will generally range from about 1000:1 to about 1:1000, preferably about 200:1 to about 1:200. Combinations of a compound of the present invention and other active ingredients will generally also be within the aforementioned range, but in each case, an effective dose of each active ingredient should be used.
In such combinations the compound of the present invention and other active agents may be administered separately or in conjunction. In addition, the administration of one element may be prior to, concurrent to, or subsequent to the administration of other agent(s).
The compounds of the present invention may be administered by oral, parenteral (e.g., intramuscular, intraperitoneal, intravenous, ICV, intracisternal injection or infusion, subcutaneous injection, or implant), by inhalation spray, nasal, vaginal, rectal, sublingual, or topical routes of administration and may be formulated, alone or together, in suitable dosage unit formulations containing conventional non-toxic pharmaceutically acceptable carriers, adjuvants and vehicles appropriate for each route of administration. In addition to the treatment of warm-blooded animals such as mice, rats, horses, cattle, sheep, dogs, cats, monkeys, etc., the compounds of the invention are effective for use in humans.
The pharmaceutical compositions for the administration of the compounds of this invention may conveniently be presented in dosage unit form and may be prepared by any of the methods well known in the art of pharmacy. All methods include the step of bringing the active ingredient into association with the carrier which constitutes one or more accessory ingredients. In general, the pharmaceutical compositions are prepared by uniformly and intimately bringing the active ingredient into association with a liquid carrier or a finely divided solid carrier or both, and then, if necessary, shaping the product into the desired formulation. In the pharmaceutical composition the active object compound is included in an amount sufficient to produce the desired effect upon the process or condition of diseases. As used herein, the term “composition” is intended to encompass a product comprising the specified ingredients in the specified amounts, as well as any product which results, directly or indirectly, from combination of the specified ingredients in the specified amounts.
The pharmaceutical compositions containing the active ingredient may be in a form suitable for oral use, for example, as tablets, troches, lozenges, aqueous or oily suspensions, dispersible powders or granules, emulsions, hard or soft capsules, or syrups or elixirs. Compositions intended for oral use may be prepared according to any method known to the art for the manufacture of pharmaceutical compositions and such compositions may contain one or more agents selected from the group consisting of sweetening agents, flavoring agents, coloring agents and preserving agents in order to provide pharmaceutically elegant and palatable preparations. Tablets contain the active ingredient in admixture with non-toxic pharmaceutically acceptable excipients which are suitable for the manufacture of tablets. These excipients may be for example, inert diluents, such as calcium carbonate, sodium carbonate, lactose, calcium phosphate or sodium phosphate; granulating and disintegrating agents, for example, corn starch, or alginic acid; binding agents, for example starch, gelatin or acacia, and lubricating agents, for example magnesium stearate, stearic acid or talc. The tablets may be uncoated or they may be coated by known techniques to delay disintegration and absorption in the gastrointestinal tract and thereby provide a sustained action over a longer period. For example, a time delay material such as glyceryl monostearate or glyceryl distearate may be employed. They may also be coated by the techniques described in the U.S. Pat. Nos. 4,256,108; 4,166,452; and 4,265,874 to form osmotic therapeutic tablets for control release.
Formulations for oral use may also be presented as hard gelatin capsules wherein the active ingredient is mixed with an inert solid diluent, for example, calcium carbonate, calcium phosphate or kaolin, or as soft gelatin capsules wherein the active ingredient is mixed with water or an oil medium, for example peanut oil, liquid paraffin, or olive oil.
Aqueous suspensions contain the active materials in admixture with excipients suitable for the manufacture of aqueous suspensions. Such excipients are suspending agents, for example sodium carboxymethylcellulose, methylcellulose, hydroxypropylmethylcellulose, sodium alginate, polyvinyl-pyrrolidone, gum tragacanth and gum acacia; dispersing or wetting agents may be a naturally-occurring phosphatide, for example lecithin, or condensation products of an alkylene oxide with fatty acids, for example polyoxyethylene stearate, or condensation products of ethylene oxide with long chain aliphatic alcohols, for example heptadecaethyleneoxycetanol, or condensation products of ethylene oxide with partial esters derived from fatty acids and a hexitol such as polyoxyethylene sorbitol monooleate, or condensation products of ethylene oxide with partial esters derived from fatty acids and hexitol anhydrides, for example polyethylene sorbitan monooleate. The aqueous suspensions may also contain one or more preservatives, for example ethyl or n-propyl p-hydroxybenzoate, one or more coloring agents, one or more flavoring agents, and one or more sweetening agents, such as sucrose or saccharin.
Oily suspensions may be formulated by suspending the active ingredient in a vegetable oil, for example arachis oil, olive oil, sesame oil or coconut oil, or in a mineral oil such as liquid paraffin. The oily suspensions may contain a thickening agent, for example beeswax, hard paraffin or cetyl alcohol. Sweetening agents such as those set forth above, and flavoring agents may be added to provide a palatable oral preparation. These compositions may be preserved by the addition of an anti-oxidant such as ascorbic acid.
Dispersible powders and granules suitable for preparation of an aqueous suspension by the addition of water provide the active ingredient in admixture with a dispersing or wetting agent, suspending agent and one or more preservatives. Suitable dispersing or wetting agents and suspending agents are exemplified by those already mentioned above. Additional excipients, for example sweetening, flavoring and coloring agents, may also be present.
The pharmaceutical compositions of the invention may also be in the form of oil-in-water emulsions. The oily phase may be a vegetable oil, for example olive oil or arachis oil, or a mineral oil, for example liquid paraffin or mixtures of these. Suitable emulsifying agents may be naturally-occurring gums, for example gum acacia or gum tragacanth, naturally-occurring phosphatides, for example soy bean, lecithin, and esters or partial esters derived from fatty acids and hexitol anhydrides, for example sorbitan monooleate, and condensation products of the said partial esters with ethylene oxide, for example polyoxyethylene sorbitan monooleate. The emulsions may also contain sweetening and flavoring agents.
Syrups and elixirs may be formulated with sweetening agents, for example glycerol, propylene glycol, sorbitol or sucrose. Such formulations may also contain a demulcent, a preservative and flavoring and coloring agents.
The pharmaceutical compositions may be in the form of a sterile injectable aqueous or oleagenous suspension. This suspension may be formulated according to the known art using those suitable dispersing or wetting agents and suspending agents which have been mentioned above. The sterile injectable preparation may also be a sterile injectable solution or suspension in a non-toxic parenterally-acceptable diluent or solvent, for example as a solution in 1,3-butanediol. Among the acceptable vehicles and solvents that may be employed are water, Ringer's solution and isotonic sodium chloride solution. In addition, sterile, fixed oils are conventionally employed as a solvent or suspending medium. For this purpose any bland fixed oil may be employed including synthetic mono- or diglycerides. In addition, fatty acids such as oleic acid find use in the preparation of injectables.
The compounds of the present invention may also be administered in the form of suppositories for rectal administration of the drug. These compositions can be prepared by mixing the drug with a suitable non-irritating excipient which is solid at ordinary temperatures but liquid at the rectal temperature and will therefore melt in the rectum to release the drug. Such materials are cocoa butter and polyethylene glycols.
For topical use, creams, ointments, jellies, solutions or suspensions, etc., containing the compounds of the present invention are employed. (For purposes of this application, topical application shall include mouthwashes and gargles.)
The pharmaceutical composition and method of the present invention may further comprise other therapeutically active compounds as noted herein which are usually applied in the treatment of the above mentioned pathological conditions.
In the treatment or prevention of conditions which require inhibition of stearoyl-CoA delta-9 desaturase enzyme activity an appropriate dosage level will generally be about 0.01 to 500 mg per kg patient body weight per day which can be administered in single or multiple doses. Preferably, the dosage level will be about 0.1 to about 250 mg/kg per day; more preferably about 0.5 to about 100 mg/kg per day. A suitable dosage level may be about 0.01 to 250 mg/kg per day, about 0.05 to 100 mg/kg per day, or about 0.1 to 50 mg/kg per day. Within this range the dosage may be 0.05 to 0.5, 0.5 to 5 or 5 to 50 mg/kg per day. For oral administration, the compositions are preferably provided in the form of tablets containing 1.0 to 1000 mg of the active ingredient, particularly 1.0, 5.0, 10.0, 15.0, 20.0, 25.0, 50.0, 75.0, 100.0, 150.0, 200.0, 250.0, 300.0, 400.0, 500.0, 600.0, 750.0, 800.0, 900.0, and 1000.0 mg of the active ingredient for the symptomatic adjustment of the dosage to the patient to be treated. The compounds may be administered on a regimen of 1 to 4 times per day, preferably once or twice per day.
When treating or preventing diabetes mellitus and/or hyperglycemia or hypertriglyceridemia or other diseases for which compounds of the present invention are indicated, generally satisfactory results are obtained when the compounds of the present invention are administered at a daily dosage of from about 0.1 mg to about 100 mg per kilogram of animal body weight, preferably given as a single daily dose or in divided doses two to six times a day, or in sustained release form. For most large mammals, the total daily dosage is from about 1.0 mg to about 1000 mg, preferably from about 1 mg to about 50 mg. In the case of a 70 kg adult human, the total daily dose will generally be from about 7 mg to about 350 mg. This dosage regimen may be adjusted to provide the optimal therapeutic response.
It will be understood, however, that the specific dose level and frequency of dosage for any particular patient may be varied and will depend upon a variety of factors including the activity of the specific compound employed, the metabolic stability and length of action of that compound, the age, body weight, general health, sex, diet, mode and time of administration, rate of excretion, drug combination, the severity of the particular condition, and the host undergoing therapy.
Alk=alkyl
APCI=atmospheric pressure chemical ionization
Ar=aryl
Boc=tert-butoxycarbonyl
br=broad
d=doublet
DBU=1,8-diazabicyclo[5.4.0]undec-7-ene
DAST=diethylaminosulfur trifluoride
Deoxofluor®=bis(2-methoxyethyl)aminosulfur trifluoride
DIBAL-H=diisobutylaluminum hydride
DMSO=dimethyl sulfoxide
ESI=electrospray ionization
EtOAc=ethyl acetate
m=multiplet
m-CPBA=3-chloroperoxybenzoic acid
MeOH=methyl alcohol
MS=mass spectroscopy
NaHMDS=sodium bis(trimethylsilyl)amide
NMR=nuclear magnetic resonance spectroscopy
PG=protecting group
rt=room temperature
s=singlet
t=triplet
THF=tetrahydrofuran
TsOH=toluene-4-sulfonic acid
The compounds of structural formula I can be prepared according to the procedures of the following Schemes and Examples, using appropriate materials and are further exemplified by the following specific examples. The compounds illustrated in the examples are not, however, to be construed as forming the only genus that is considered as the invention. The Examples further illustrate details for the preparation of the compounds of the present invention. Those skilled in the art will readily understand that known variations of the conditions and processes of the following preparative procedures can be used to prepare these compounds.
Compounds of structural formula I wherein X is C can be prepared by Method A. An appropriately substituted and N-protected-4-hydroxypiperidine (1) is first coupled to an ArOH, ArSH or ArNH(acyl) unit by a Mitsunobu reaction (see Tanaka, N.; Goto, R.; Ito, R.; Hayakawa, M.; Ogawa, T.; Fujimoto, K. Chem. Pharm. Bull. 1998, 46, 639-646; Fletcher, S. R.; Burkamp, F.; Blurton, P.; Cheng, S. K. F.; Clarkson, R.; O'Connor, D.; Spinks, D.; Tudge, M.; Niel, M. B.; Patel, S.; Chapman, K.; J. Med. Chem. 2002, 45, 492-503; Ohno, K.-I.; Fukushima, T.; Santa, T.; Waizumi, N.; Tokuyama, H.; Masako, M.; Imai, K.; Anal. Chem. 2002, 74, 43914396). The piperidine nitrogen protecting group (PG) is then cleaved to give 3 which is coupled to a brominated-heteroaromatic ester 4 via a nucleophillic-aromatic-substitution reaction to give 5. The ester functionality in 5 is converted into an aldehyde by a two step sequence involving treatment with a reducing agent such as DIBAL-H or LiAlH4 followed by oxidation of the resulting alcohol with MnO2 or alternatively a DMSO derived oxidant (i.e. Swern, Moffatt). The aldehyde can alternatively be converted to a ketone by reaction with an organometallic reagent followed by oxidation in a similar manner. Knoevenagel condensation of aldehyde or ketone 6 with a 2-substituted-malonic acid 7 leads to the unsaturated acid 8 which is converted to the corresponding acylazide 9 through the intermediacy of an acid chloride or mixed anhydride (as shown). Curtius rearrangement of the α,β-unsaturated-acylazide under thermal conditions generates an interconverting mixture of cis- and trans-α,β-unsaturated-isocyanates. The cis-isomer subsequently participates in an intramolecular electrophillic-aromatic-substitution reaction (see Shafiee, A.; Ghazar, H. J. Heterocycl. Chem. 1986, 23, 1171-1173) to give compounds of the present invention represented by structure 10. For Y=S, further elaboration of 10 to the corresponding sulfoxide (p=1) and sulfone (p=2) derivatives (11) is accomplished by oxidation with a suitable oxidizing agent, such as m-CPBA.
Piperidine 3 from above can be elaborated to 12 according to documented literature procedures (Z=S, O, N; see Ried W.; Kuhnt D. Liebigs. Ann. Chem. 1986, 780-784; McCarty, C. G. et al., J. Org. Chem. 1970, 35, 2067-2069; Gante J.; Mohr G. Chem. Ber. 1975, 108, 174-180, respectively). Treatment of 12 with a suitable base and solvent combination such as triethylamine in methanol affords the 5-membered heteroaromatic ring in 13 via an intramolecular attack of a resonance-stabilized carbanion (G=nitrile, ester or amide when Z=S; nitrile when Z=O or NR18) onto the carbon of the cyanamide (see Ried W.; Kuhnt D. Liebigs Ann. Chem. 1986, 780-784). When G=CONH2, subsequent reaction with an orthoester in the presence of an acid catalyst such as TsOH affords compounds of the present invention denoted by 14. Alternatively, diazotization of the amino group in 13 with nitrous acid and concomitant attack of the amide nitrogen onto the terminal diazonium nitrogen (see Ried W.; Kuhnt D. Liebig Ann. Chem. 1986, 780-784) leads to compounds of the present invention represented by 15. When G=CN, reaction of 13 with acetimidates such as 16 gives compounds of the present invention embodied by 17.
Compounds of the present invention wherein X-Y is CR17CR1R2 can be prepared as outlined below in conjunction with the transformations outlined in Methods A and B. Compounds wherein R1, R2, and R17 are hydrogen can be accessed from intermediate 20 which is prepared by a Wittig reaction between 19 and piperidone 18 followed by reduction of the resulting alkene with H2 in the presence of a suitable catalyst such as Pd on charcoal. Compounds wherein X-Y is CH—C(═O) can be generated from 22 which is prepared by reaction of 18 with alkoxysubstituted-Wittig reagent 21 followed by acid catalyzed hydrolysis of the resulting enol-ether. Intermediate 22 can be further elaborated to chlorinated derivative 23 via treatment of the corresponding hydrazone with CuCl2 (see Takeda, T.; Sasaki, R.; Satoshi, Y.; Fujiwara, T. Tetrahedron 1997, 53, 557-566) or to fluorinated derivative 24 by treatment of 22 with DAST or Deoxofluor®. Alkenyl-analogs such as 26 are prepared from 22 using Wittig reagents of the type represented by 25. Olefin 26 can also be transformed into the corresponding spiro-cyclopropane 27 by a Simmons-Smith reaction.
Introduction of an alcohol or fluorine substituent at the 4-position of the piperidine subunit can be accomplished as outlined below. Epoxide 28 is reacted with a suitable organometallic reagent to give alcohol 29 (see Castro, J. L.; Collins, I.; Russell, M. G. N.; Watt, A. P.; Sohal, B. et al. J. Med. Chem. 1998, 41, 2667-2670). Subsequent treatment with DAST or Deoxofluor® yields the fluorinated intermediate 30. Utilization of 29 or 30 in Method A or B yields compounds of the present invention.
Compounds wherein X-Y is CF—CF2 can be prepared as follows. The enol-ether product of the Wittig reaction between compounds 18 and 21 can be converted to the corresponding epoxide 31a by treatment with an appropriate oxidizing agent such as m-CPBA. Hydrolytic opening of the epoxide yields α-hydroxyketone 32. Intermediate 32 can alternatively be prepared from 31b in an analogous manner. Treatment of 32 with DAST or Deoxofluor® leads to trifluorinated intermediate 33. Compounds 32 and 33 can then be converted to compounds of the present invention according to Methods A and B.
Compounds wherein X-Y is CH—CHF can be prepared as follows. Addition of an aryl-Grignard or aryl-lithium reagent to aldehyde 34 gives alcohol 35 and subsequently fluoro-compound 36 through treatment with DAST or Deoxofluor®. Intermediates 35 and 36 are then transformed into compounds of the present invention using Method A or B.
Piperidine 3 can be converted, using standard methods, to the corresponding urea (Z=O) (phosgene and ammonia), thiourea (Z=S) (thiophosgene and ammonia; also see Mohanta, P. K.; Dhar, S.; Samal, S. K.; Junjappa, H. Tetrahedron 2000, 56, 629-637) or guanidine (Z=NH) (see Ghosh, A. K.; Hol W. G. J.; Fan, E. J. Org. Chem. 2001, 66, 2161-2164; Lazar, J.; Bernath, G. J. Heterocycl. Chem. 1990, 27, 1885-1892) derivatives 37. Subsequent Hantzsch-type reaction with the appropriate bromobarbituric acid yields compound 38 of the present invention.
Wherein X-Y is N—C(O) or NS(O)2, a t-butyloxycarbonyl (Boc) or benzyloxycarbonyl (Cbz) protected piperazine 39 is reacted with an aroyl halide in the presence of a base such as a tertiary amine, alkali metal carbonate, and alkali metal hydroxide. The intermediate is then deprotected in a standard manner to give the desired amine 40 for further elaboration into the final compounds as described in Methods A-G.
Piperidine 3 can participate in an SNAr reaction with heterocycles such as 41 and 42 to afford 43 and 45 respectively. Reaction of 43 or 45 with appropriately substituted hydrazines affords compounds of the present invention embodied by 44 and 46.
An appropriately substituted heteroaryl halide 1 is reacted with an appropriately substituted cyclic amine 2 in the presence of a base such as 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU), triethylamine or an alkali metal (K, Na, Cs) carbonate in a solvent such as N,N-dimethylformamide (DMF), ethanol, 2-methoxyethanol, and aqueous mixtures thereof at a temperature range of about room temperature to about refluxing temperature. Extractive work up and purification by flash column chromatography or precipitation of the product by the addition of saturated sodium bicarbonate solution or water gives desired condensed product 3.
The following examples are provided to illustrate the invention and are not to be construed as limiting the invention in scope in any manner.
Diethyl azodicarboxylate (18.9 mL, 120 mmol) was added dropwise to a 0° C. solution of tert-butyl-4-hydroxypiperidine-1-carboxylate (20.13 g, 100 mmol), 2-trifluoromethylphenol (17.83 g, 110 mmol) and triphenylphosphine (31.44 g. 120 mmol) in THF (300 mL). The mixture was then warmed to rt and stirred for 16 h before being concentrated and partitioned between ether and water. The ether phase was washed with 2 M NaOH and water, dried over Na2SO4 and concentrated. The residue was then suspended in a mixture of ether and hexanes (35/65) and filtered to remove most of the triphenylphosphine oxide by-product. The filtrate was concentrated and the residue was subjected to flash chromatography on silica gel eluting with 35/65 ether/hexanes to afford the title compound as a colorless solid.
A solution of tert-butyl 4-[2-(trifluoromethyl)phenoxy]piperidine-1-carboxylate (28.65 g, 83.0 mmol) in CH2Cl2 (200 μL) was cooled to 0° C. and treated with trifluoroacetic acid (25.5 mL, 330 mmol) with stirring at rt for 10 h. The reaction mixture was then concentrated and the residue was taken up in ethyl acetate, washed with 2 M NaOH and brine, and the organic phase was dried over Na2SO4. Concentration in vacuo and flash chromatography on silica gel eluting with Jan. 9, 1990 NH4OH/MeOH/CH2Cl2 gave the title compound as a faint-yellow syrup.
A solution of 4-[2-{trifluoromethyl}phenoxy]piperidine (5.70 g, 23.3 mmol), ethyl 2-bromothiazole-4-carboxylate (5.50 g, 23.3 mmol) and DBU (7.0 mL, 46.6 mmol) in THF (70 mL) was heated at 80° C. for 20 h. The mixture was then poured into water and extracted with ethyl acetate. The organic phase was washed with water, dried (Na2SO4) and concentrated, and the residue was subjected to flash chromatography on silica gel eluting with 3/7 EtOAc/hexanes to yield the title compound as a yellow oil.
DIBAL-H (3.3 mL, 1.5 M in toluene, 5.0 mmol) was added dropwise to a −70° C. solution of ethyl 2-{4-[2-{trifluoromethyl}phenoxy]piperidin-1-yl}-1,3-thiazole-4-carboxylate (1.22 g, 3.05 mmol) in a mixture of THF and CH2Cl2 (1:1, 50 mL). After 2 h at this temperature, methanol (5 mL) was added dropwise and the reaction vessel contents were warmed slowly to rt and then partitioned between water and ethyl acetate. The organic phase was washed with water, dried (Na2SO4) and concentrated. Flash chromatography of the residue on silica gel eluting with an increasing proportion of ethyl acetate in hexanes (2/3 to 1/1 to 7/3) afforded the title compound as a yellow oil.
2-{4-[2-{Trifluoromethyl}phenoxy]piperidin-1-yl}-1,3-thiazole-4-carbaldehyde (558 mg, 1.56 mmol), malonic acid (180 mg, 1.72 mmol) and piperidine (20 μL, 10 mol %) were heated together in pyridine (1.6 mL) at reflux for 0.5 h. Concentration in vacuo and flash on silica eluting with a mixture consisting of ethyl acetate/hexanes (1:1) and 0.5% v/v acetic acid provided the title compound as a yellow oil.
Ethyl chlorocarbamate (65 μL, 0.64 mmol) was added to a mixture of 3-(2-{4-[2-(trifluoromethyl)phenoxy]piperidin-1-yl}-1,3-thiazol-4-yl)acrylic acid (240 mg, 0.61 mmol) and Et3N (110 μL, 0.73 mmol) in acetone (2.5 mL) at 0° C. After stirring 15 min at this temperature, a solution of NaN3 (72 mg, 1.1 mmol) in water (2 mL) was introduced dropwise followed by continued stirring at 0° C. for 1 h. The reaction mixture was then partitioned between ethyl acetate and water and the layers were separated. The organic phase was dried over Na2SO4 and concentrated, and the residue was subjected to flash chromatography on silica eluting with 1/3 ethyl acetate/hexanes to give the title compound as a yellow oil.
3-(2-{4-[2-(Trifluoromethyl)phenoxy]piperidin-1-yl}-1,3-thiazol-4-yl)acryloyl azide (140 mg, 0.34 mmol) was heated at 200° C. in diphenyl ether (2 mL) for 2 h. The reaction mixture was then loaded onto silica gel and eluted with 7/3 ethyl acetate/hexanes followed by 9/1 ethyl acetate/methanol to yield the title compound as a faint-yellow solid. 1H NMR (500 MHz, d6-acetone): δ 8.80 (1H, d), 8.51 (1H, dd), 7.68-7.60 (1H, m), 7.35 (1H, dd), 7.29 (1H, t), 7.13 (1H, t), 5.06 (1H, m), 4.05-3.84 (2H, m), 3.78-3.61 (2H, m), 2.40-1.89 (4H, m) ppm. MS (+ESI) 380.0 [M+H]+.
4-[2-{Trifluoromethyl}phenoxy]piperidine (1.12 g, 4.57 mmol, from Step 1 of Example 1) and dimethyl N-cyanodithioiminocarbonate (670 mg, 4.60 mmol) were heated together at reflux in ethanol (1.5 mL) for 30 min. The mixture was then concentrated in vacuo to afford the title compound as a thick yellow syrup.
Triethylamine (2.0 mL, 15 mmol) was added to a mixture of methyl N-cyano-[2-(trifluoromethyl)phenoxy]piperidine-1-carbimidothioate (1.56 g, 4.57 mmol), 2-mercaptoacetamide (4.2 mL, 4.6 mmol, 10 wt % in methanolic ammonia) and solution was left to stand at rt overnight after thorough mixing by swirling. The mixture was then cooled to 0° C. and filtered. The solid that was collected was washed with ice cold methanol and dried under vacuum to afford the title compound as a colorless powder.
A solution of NaNO2 (90 mg, 1.3 mmol) in H2O (0.30 mL) was added dropwise at 0° C. to a slurry of 4-amino-2-{4-[2-(trifluoromethyl)phenoxy]piperidin-1-yl}-1,3-thiazole-5-carboxamide (110 mg, 260 μmol) in 6 M HCl (1.5 mL). After stirring at 0° C. for 10 min, then at rt for 1 h, the solids produced during the reaction were removed by filtration and loaded onto silica gel. Flash chromatography eluting with 1/3 acetone/benzene gave the title compound as a yellow solid. 1H NMR (500 MHz, d6-acetone): δ 14.05 (1H, br s), 7.68-7.64 (2H, m), 7.41 (1H, d), 7.14 (1H, t), 5.11 (1H, m), 3.93 (4H, m), 2.28-2.20 (2H, m), 2.15-2.05 (2H, overlapped m) ppm. MS (+ESI) 398.0 [M+H]+.
4-Amino-2-{4-[2-(trifluoromethyl)phenoxy]piperidin-1-yl}-1,3-thiazole-5-carboxamide (from Step 2 of Example 2) (100 mg, 260 μmol) and trimethylorthoformate (3 mL) were heated together at reflux in the presence of TsOH.H2O (5 mg, 10 mol %) for 30 min. The mixture was then cooled to rt and the solid produced by the reaction was collected by filtration and washed with ether to afford the title compound as a colorless powder. 1H NMR (400 MHz, d6-acetone): δ 11.10 (1H, br s), 8.13 (1H, s), 7.66 (2H, m), 7.41 (1H, d), 7.14 (1H, t), 5.08 (1H, m), 3.91-3.83 (4H, m), 2.25-2.19 (2H, m), 2.11-2.04 (2H, overlapped m) ppm. MS (−APCI) 395.0 [M−H]−.
A solution of thioacetonitrile (0.8 M in methanol, 10 ml, 8 mmol; for preparation of this reagent see Gaumont, A. C.; Wazneh, L.; Denis, J. M. Tetrahedron 1991, 47, 492740) and methyl N-cyano-[2-(trifluoromethyl)phenoxy]piperidine-1-carbimidothioate (from Step 1 of Example 2) (1.1 g, 3.2 mmol) was treated with Et3N (2.2 mL, 22 mmol) with stirring at rt for 1 h. The mixture was then poured into water (40 mL) and the resulting solid was collected by filtration and washed well with water. Recrystallization from ethanol/water yielded the title compound as a light-brown powder.
A solution of 4-amino-2-{4-[2-(trifluoromethyl)phenoxy]piperidin-1-yl}-1,3-thiazole-5-carbonitrile (110 mg, 300 mmol) was heated in acetamide (0.5 mL) at 210° C. in a sealed tube under microwave irradiation for 20 min. The mixture was then poured into water and extracted with 1:1 EtOAc/THF. The organic extract was washed with brine, dried (Na2SO4/MgSO4) and concentrated. Flash chromatography of the residue on silica gel eluting with Jan. 9, 1990 NH4OH/MeOH/CH2Cl2 provided the title compound as a faint-yellow powder. 1H NMR (500 MHz, d3-acetonitrile): δ 7.65-7.59 (2H, m), 7.28 (1H, d), 7.10 (1H, t), 6.36 (2H, br s), 4.93 (1H, m), 3.84-3.77 (4H, m), 2.13-2.07 (2H, m), 1.96-1.91 (2H, m) ppm. MS (+ESI) 396.3 [M+H]+.
A mixture of 2-(trifluoromethyl)benzyl bromide (12.25 g, 51.25 mmol) and triphenylphosphine (13.44 g, 51.30 mmol) were heated together in refluxing acetonitrile (50 mL) for 24 h. The acetonitrile was removed in vacuo and the residue was triturated with ether to afford the title compound as a colorless powder.
A solution of NaHMDS (1 M in THF, 22 mL) was added dropwise to a 0° C. slurry of triphenyl[2-(trifluoromethyl)benzyl]phosphonium bromide (10 g, 20 mmol) in THF (20 mL). The resulting orange slurry was stirred at rt for 20 min prior to the introduction of N-Boc-4-piperidone (4.40 g, 22 mmol) as a solid followed by heating at 50° C. for 24 h. Water and 2 M HCl were added at 0° C. and the mixture was extracted with ether. The organic phase was washed with water, brine, saturated aqueous sodium bicarbonate solution, and dried (Na2SO4/MgSO4). Concentration in vacuo and flash chromatography on silica eluting with 1/4 ether/hexanes provided the title compound as a colorless oil.
A mixture of tert-butyl 4-[2-(trifluoromethyl)benzylidene]piperidine-1-carboxylate (3.33 g, 9.74 mmol) and 10% palladium on charcoal (2.10 g) in ethyl acetate (60 mL) was stirred under an atmosphere of hydrogen for 3 h. The catalyst was removed by filtration through Celite® and the filtrate was concentrated to yield the title compound as a colorless syrup.
Trifluoroacetic acid (20 mL) was added to a 0° C. solution of tert-butyl 4-[2-(trifluoromethyl)benzyl]piperidine-1-carboxylate (3.31 g, 9.62 mmol) in CH2Cl2 (20 mL) with stirring at rt for 1 h. Concentration in vacuo afforded the crude amine that was taken up in ethyl acetate and washed with 10% aqueous Na2CO3 solution, brine and dried (Na2SO4). Evaporation of the solvents and trituration with ether gave the title compound as a colorless powder.
The title compound was prepared from 4-[2-(trifluoromethyl)benzyl]piperidine according to the procedures detailed in Example 2 (step 2) and Example 3 to afford the title compound as a colorless solid. 1H NMR (500 MHz, d6-DMSO): δ 12.37 (1H, s), 8.08 (1H, s), 7.71 (1H, d), 7.64 (1H, t), 7.51 (1H, d), 7.45 (1H, t), 4.05 (2H, s), 3.15 (2H, br t), 2.74 (2H, br d), 1.94 (1H, m), 1.73-1.65 (2H, br d), 1.36-1.28 (2H, m) ppm. MS (+ESI) 395.1 [M+H]+.
A mixture of 4-[2-(trifluoromethyl)phenoxy]piperidine (1-1) (6.0 g, 24.5 mmol), dimethyl N-cyanodithioiminocarbonate (1-2) (3.75 g, 25.6 mmol) in ethanol (24 mL) was stirred at 65° C. for 2 h. The volatile materials were removed under reduced pressure and the resulting yellow syrup was dried under high vacuum.
The methyl N-cyano-4-[2-(trifluoromethyl)phenoxy]piperidine-1-carbimidothioate from step 1 (1-3) (8.4 g, 24.5 mmol) was dissolved in a solution of 2-mercaptoacetamide in methanol (30 mL of 10 g/100 mL, 32.9 mmol). To this mixture, triethylamine was added dropwise over 10 min, and the reaction was stirred at rt for 2 h. The mixture was cooled in an ice bath to promote crystallisation with continued stirring. It was then brought back to rt and water (2 mL) was added dropwise and stirred at rt overnight. The resulting white solid was collected by filtration, washed with methanol/water (2:1) and dried under high vacuum to give the title compound as a white solid. MS (+ESI) 386.9 [M+H]+.
To a solution of 4-amino-2-{4-[2-(trifluoromethyl)phenoxy]piperidin-1-yl}-1,3-thiazole-5-carboxamide from step 2 (1-4) (1.01 g, 2.6 mmol) in acetic acid (4 mL) was slowly added acetyl chloride (0.4 mL) and the resulting mixture was stirred at 120° C. overnight. The reaction was allowed to cool to rt and it was partitioned between EtOAc and half-saturated NaHCO3, dried over Na2SO4 and concentrated. The crude product was swished in EtOAc and diethyl ether (1:1, 50 mL) to give the title compound as a light yellow solid. 1H NMR (400 MHz, d6-Acetone): δ 11.2 (1H, br. s), 7.67 (2H, m), 7.40 (1H, d), 7.12 (1H, dd), 5.06 (1H, m), 3.82 (4H, m), 2.43 (3H, s), 2.20 (2H, m), 2.0 (2H, m) ppm. MS (+APCI) 411.1 [M+H]+.
A mixture of 4-amino-2-{4-[2-(trifluoromethyl)phenoxy]piperidin-1-yl}-1,3-thiazole-5-carboxamide from Step 2 of Example 6 (0.40 g, 1.04 mmol) with chloroacetic acid (2.56 g) was warmed to 80° C. to melt the chloroacetic acid and give a homogeneous solution to which was slowly added chloroacetyl chloride (0.17 mL, 2.1 mmol) and the resulting mixture was stirred at 130° C. for 5 h. The reaction was allowed to cool to rt and it was partitioned between EtOAc and half-saturated NaHCO3, dried over Na2SO4 and concentrated. The crude product was swished and sonicated in EtOAc (7 mL), filtered, and dried to give the title compound as a light beige solid. 1H NMR (400 MHz, d6-Acetone): δ 11.42 (1H, br. s), 7.64 (2H, m), 7.39 (1H, d), 7.12 (1H, dd), 5.08 (1H, m), 4.62 (2H, s), 3.86 (4H, m), 2.20 (2H, m), 2.02 (2H, m) ppm. MS (+ESI) 445.0 (M+1).
To a solution of 5-(chloromethyl)-2-{4-[2-(trifluoromethyl)phenoxy]piperidin-1-yl}[1,3]thiazolo[4,5-d]pyrimidin-7(6H)-one (160 mg, 0.36 mmol) in DMF (1 mL) was added sodium acetate (107 mg, 1.3 mmol). The resulting mixture was stirred at 70° C. for 3 h. It was then partitioned between EtOAc and half-saturated NaHCO3, the organic layer was dried over Na2SO4 and concentrated. This crude product was purified by flash chromatography using a gradient of ethanol in EtOAc going from 0 to 10% to give the title compound as a white solid. 1H NMR (400 MHz, d6-Acetone): δ 11.22 (1H, br. s), 7.67 (2H, m), 7.40 (1H, d), 7.12 (1H, dd), 5.08 (2H+1H, s+m), 3.86 (4H, m), 2.20 (2H, m), 2.15 (3H, s), 2.03 (2H, m) ppm. MS (+APCI) 469.2 (M+1).
To a solution of (7-oxo-2-{4-[2-(trifluoromethyl)phenoxy]piperidin-1-yl}-6,7-dihydro[1,3]thiazolo[4,5-d]pyrimidin-5-yl)methyl acetate (55 mg, 0.12 mmol) in ethanol (1.0 mL) was added a freshly prepared solution of sodium ethoxide in ethanol (0.1 mL of 0.69M, 0.069 mmol). The resulting solution was stirred at room temperature overnight. It was then partitioned between EtOAc and NH4OAc buffer, the organic layer was dried over Na2SO4, and concentrated. This crude product was purified by flash chromatography using a gradient of ethanol in EtOAc going from 0 to 15% to give the title compound as a white solid. 1H NMR (400 MHz, d6-Acetone): δ 11.60 (1H, br. s), 7.67 (2H, m), 7.43 (1H, d), 7.12 (1H, dd), 5.51 (1H, br. S), 5.04 (1H, m), 4.46 (2H, m), 3.81 (4H, m), 2.16 (2H, m), 1.97 (2H, m) ppm. MS (+ESI) 427.0 (M+1).
To a solution of 5-(chloromethyl)-2-{4-[2-(trifluoromethyl)phenoxy]piperidin-1-yl}[1,3]thiazolo[4,5-d]pyrimidin-7(6H)-one (200 mg, 0.45 mmol) in DMSO (3.0 mL) was added sodium cyanide (99 mg, 2.0 mmol). The resulting solution was stirred at room temperature overnight. It was then partitioned between EtOAc and half-saturated aqueous NaHCO3, the organic layer was dried over Na2SO4, and concentrated. This crude product was purified by flash chromatography using a gradient of EtOAc in hexane going from 80 to 100% to give the title compound as a light yellow solid. 1H NMR (400 MHz, d6-Acetone): δ 11.39-11.34 (b, 1H), 7.67-7.60 (m, 2H), 7.39 (d, 1H), 7.16-7.10 (t, 1H), 5.10-5.04 (m, 1H), 4.19-4.15 (m, 2H), 3.91-3.79 (m, 4H), 2.25-2.16 (m, 2H), 2.05-1.98 (m, 2H). MS (+ESI) 436.0 (M+1).
(7-Oxo-2-{4-[2-(trifluoromethyl)phenoxy]piperidin-1-yl}-6,7-dihydro[1,3]thiazolo[4,5-d]pyrimidin-5-yl)acetonitrile (165 mg, 0.38 mmol) and azidotributyltin (0.13 mL, 0.47 mmol) in toluene (1.0 mL) were mixed and heated at 130° C. for 17 h with only a needle connected to an oil bubbler to allow the evaporation of the solvent. The resulting brown residue was partitioned between EtOAc and 1N HCl, the organic layer was extracted with saturated aqueous NaHCO3 which was washed with EtOAc. The aqueous layer was carefully acidified using 10N HCl and then extracted with EtOAc. The organic layer was dried over Na2SO4 and concentrated. The resulting amber gum was diluted with EtOAc (3 mL) and sonicated for 1 min. The resulting light-beige solid was collected by vacuum filtration. The 400 MHz NMR spectrum was recorded in a 1:10 DMSO-d6/acetone-d6 mixture: δ 7.64 (t, 2H), 7.40 (d, 1H), 7.12 (t, 1H), 5.05-5.02 (m, 1H), 4.43 (s, 2H), 3.82-3.77 (m, 4H), 2.19-2.11 (m, 2H), 1.98-1.92 (m, 2H). MS (+APCI) 479.1 (M+1).
A mixture of 8-bromoadenine (150 mg, 0.7 mmol), 4-[2-(trifluoromethyl)phenoxy]-piperidine (0.2 mL, 0.84 mmol) and triethylamine (0.12 mL, 0.84 mmol) in 2-methoxymethanol/water (4:1, 2.3 mL) was heated at 130° C. for 16 h. The volatiles were evaporated to one third the volume and diluted with saturated aqueous NaHCO3 (2 mL). The resulting solid was filtered and washed with water and then ethyl acetate. The product was dried under high vacuum to give the title product as a solid. 1H NMR (500 MHz, acetone-d6): δ 7.97 (s, 1H), 7.67-7.62 (m, 2H), 7.37 (d, 1H), 7.12 (d, 1H), 5.80 (s, 1H), 4.98 (s, 1H), 3.85 (d, 2H), 3.71 (s, 2H), 2.20-2.14 (m, 2H), 1.99-1.92 (m, 2H). MS (+ESI) m/z 379 (MH+).
A mixture of 2-amino-8-bromo-1,9-dihydro-6H-purin-6-one (100 mg, 0.4 mmol), 4-[2-(trifluoromethyl)phenoxy]piperidine (0.24 mL, 0.98 mmol) and triethylamine (0.1 mL, 0.65 mmol) in 2-methoxymethanol/water (4:1, 0.7 mL) was heated at 130° C. for 16 h. The volatiles were evaporated to one third the volume and diluted with saturated aqueous NaHCO3 (2 mL). The resulting solid was filtered and washed with water and then MeOH. The product was dried under high vacuum to give the title product as a solid.
1H NMR (500 MHz, DMSO-d6): δ 11.26 (s, 1H), 10.26 (s, 1H), 7.64-7.60 (m, 2H), 7.37 (d, 1H), 7.09 (t, 1H), 5.97 (s, 1H), 4.87 (s, 1H), 3.65 (s, 2H), 3.50 (s, 2H), 1.98 (s, 2H), 1.71 (s, 2H). MS (+ESI) m/z 395 (MH+).
A mixture of 8-chlorotheophylline (150 mg, 0.7 mmol), 4-[2-(trifluoromethyl)phenoxy]-piperidine (0.21 mL, 0.83 mmol) and triethylamine (0.14 mL, 1.0 mmol) in 2-methoxyethanol/water (4:1, 1.4 mL) was heated at 130° C. for 16 h. The volatiles were evaporated to one third the volume and diluted with saturated aqueous NaHCO3 (2 mL). The resulting solid was filtered and washed with water and then ether. The product was dried under high vacuum to give the title product as a solid.
1H NMR (500 MHz, acetone-d6): δ 7.64-7.58 (m, 2H), 7.34 (d, 1H), 7.09 (t, 1H), 4.99-4.95 (m, 1H), 3.87-3.81 (m, 2H), 3.77-3.71 (m, 2H), 3.43 (s, 3H), 3.23 (s, 3H), 2.17-2.10 (m, 2H), 1.96-1.90 (m, 2H). MS (+ESI) m/z 424 (MH+).
To a mixture of xanthine (100 mg, 0.66 mmol) in water (0.65 mL) was added bromine (0.05 mL, 0.99 mmol) in a 4-mL glass vial. The vial was capped and heated at 100° C. After 2 h, the mixture was cooled to room temperature, filtered and the solid washed with water and then Et2O. The product was dried under high vacuum to give the title product as a solid.
1H NMR (500 MHz, DMSO-d6): δ 14.09 (1H, s), 11.64 (1H, s), 10.91 (1H, s). MS (+ESI) m/z 232, 233 (MH+).
A mixture of 8-bromo-3,9-dihydro-1H-purine-2,6-dione (80 mg, 0.34 mmol), 4-[2-(trifluoromethyl)phenoxy]piperidine (0.10 mL, 0.41 mmol) and triethylamine (0.07 mL, 0.52 mmol) in 2-methoxyethanol/water (4:1, 0.69 mL) was heated at 130° C. for 16 h. The volatiles were evaporated to one third the volume and diluted with saturated aqueous NaHCO3 (2 mL). The resulting solid was filtered and washed with water and then ether. The product was recrystallized from MeOH/Et2O to give the title product as a solid.
1H NMR (500 MHz, CD3OD): δ 7.59-7.53 (m, 2H), 7.22 (d, 1H), 7.04 (t, 1H), 3.72-3.59 (m, 4H), 2.08-2.01 (m, 2H), 1.95-1.88 (m, 2H). MS (+ESI) m/z 396 (MH+).
As a specific embodiment of an oral composition of a compound of the present invention, 50 mg of the compound of any of the Examples is formulated with sufficient finely divided lactose to provide a total amount of 580 to 590 mg to fill a size 0 hard gelatin capsule.
While the invention has been described and illustrated in reference to specific embodiments thereof, those skilled in the art will appreciate that various changes, modifications, and substitutions can be made therein without departing from the spirit and scope of the invention. For example, effective dosages other than the preferred doses as set forth hereinabove may be applicable as a consequence of variations in the responsiveness of the human being treated for a particular condition. Likewise, the pharmacologic response observed may vary according to and depending upon the particular active compound selected or whether there are present pharmaceutical carriers, as well as the type of formulation and mode of administration employed, and such expected variations or differences in the results are contemplated in accordance with the objects and practices of the present invention. It is intended therefore that the invention be limited only by the scope of the claims which follow and that such claims be interpreted as broadly as is reasonable.
| Filing Document | Filing Date | Country | Kind | 371c Date |
|---|---|---|---|---|
| PCT/CA2006/001847 | 11/10/2006 | WO | 00 | 4/21/2008 |
| Number | Date | Country | |
|---|---|---|---|
| 60813644 | Jun 2006 | US | |
| 60736824 | Nov 2005 | US |
