WO2003090746A1 - 1,3-thiazoles as lxr modulators in the treatment of cardiovascular diseases - Google Patents

Abstract

The present invention relates to methods for treatment of certain diseases or conditions mediated by Liver X Receptor (LXR) by the administration of a composition containing as an active ingredient a compound according to Formula I. In particular, the invention relates to methods for treatment of cardiovascular diseases and atherosclerosis through the administration of a compound which modulates LXR activity.

Description

, 3-THIAZOLES AS LXR MODULATORS IN THE TREATMENT OF CARDIOVASCULAR DISEASES

FIELD OF THE INVENTION

The present invention relates to methods for treatment of certain diseases or conditions mediated by Liver X Receptor (LXR) by the administration of a composition containing as an active ingredient a compound according to Formula I. In particular, the invention relates to methods for treatment of cardiovascular diseases and atherosclerosis through the administration of a compound which modulates LXR activity.

BACKGROUND

Liver X receptors (LXRs), LXRα and LXRβ, are nuclear receptors that regulate the expression of cytochrome P450 7A (CYP7A1), and thus the metabolism of several important Iipids, including cholesterol and bile acids. LXRs were first identified as orphan members of the nuclear receptor superfamily (Song et al., Proc. Natl. Acad. Sci. 191 :10809-10813 (1994). Willy et al., Gene Deve. 9:1033-1045 (1995)). The identification of a specific class of oxidized derivatives of cholesterol (oxysterol) as ligands for the LXRs, in combination with the description of an LXR response element in the promoter of the rat cholesterol 7α-hydroxylase genes, suggested that LXRs play an important role in the regulation of cholesterol homeostasis. Additional support for this role came from the analysis of LXRα-deficient mice (LXRα-/-), which uncovered the dysregulation of the CYP7A1 gene and several other important lipid-associated genes (Peet et al., Curr. Opin. Genet. Dev. 8: 571-575 (1998)). Studies utilizing these animals confirmed the essential function of LXRα as a major sensor of dietary cholesterol and an activator of the bile acid synthetic pathway in mice. LXRα is expressed most highly in the liver and to a lesser extent in the kidney, small intestine, spleen and adrenal gland. On the contrary, LXRβ is ubiquitously expressed. Naturally occurring or synthetic oxysterols such as 22(R)- hydroxycholesterol, 24(S)-hydroxycholesterol, and 24(S),25-epoxycholesterol are believed to be transcriptional activators of LXRα and β. These oxysterols exist at concentrations that activate LXRs in tissues (e.g. liver, brain and placenta) where both cholesterol metabolism and LXR expression are high. In human monocyte-derived macrophages by cholesterol loading, it was demonstrated a dose-dependent induction of ABCA1 and ABCG1 genes, suggesting that 27-hydroxycholesterol is an endogenous ligand for LXRs (Fu et al, J. Biol. Chem. 276:38378-38387(2001 )).

LXRs bind to the ATP binding cassette transporter-1 (ABCA1) gene promoter and increases expression of the gene to result in increased ABCA1 protein. ABCA1 is a membrane bound transport protein which is involved in the regulation of cholesterol efflux from extrahepatic cells onto nascent HDL particles. Humans with mutations in the gene ABCA1 have low levels of high density lipoprotein (HDL) and a concomitant increased risk of cardiovascular diseases such as atherosclerosis, myocardial infarction and ischemic stroke (Brooks-Wilson et al, Nat. Genet. 22: 336-345 (1999), Bodzioch et al., Nat Genet. 22: 347-351 (1999); and Rust et al., Nat. Genet. 22: 352-355 (1999)). LXRα and β agonists were demonstrated to increase ABCA1 gene expression which resulted in increased HDL cholesterol, and decreased absorption of cholesterol and thereby decreased the risk of cardiovascular diseases (Sparrow et al., J. Biol. Chem. 277:10021-10027 (2002).

LXRs signaling pathways play a central role in the control of macrophage cholesterol efflux through the coordinate regulation of ABCA1 and ABCG1 and surface constituent of plasma lipoprotein apolipoprotein E (apoE) gene expression. Recently, it was demonstrated that LXR/RXR heterodimers regulate apoE transcription directly, through interaction with a conserved LXR response element present in both ME.1 and ME.2. The ability of oxysterol and synthetic ligands to regulate apoE expression in adipose tissue and peritoneal macrophages is reduced in LXRα-/- or LXRβ-/- mice and abolished in double knockouts.

LXRs also play an important role in fatty acid metabolism by activating the sterol regulatory element-binding protein-1c (SREBP-1c) gene (Tobin, et al., J. Biol. Chem. 277:10691-10697 (2002). In rodent liver and hepatoma cells, transcription of the SREBP-1c gene is stimulated by naturally occurring oxysterols, like 24(S),25-expoxycholesterol and 22(R)-hydroxycholesterol, that bind to LXRα and β. LXRs are also activated by T0901317, a synthetic nonsteroidal compound. The level of SREBP-1c mRNA declined dramatically when cultured rat hepatoma cells were treated with inhibitors of 3-hydroxy-3- methylglutaryl coenzyme reductase, which block the synthesis of endogenous LXR ligands. This inhibition was reversed when the cells were incubated with either T0901317 or mevalonate, the product of the reductase reaction. These data indicated that basal transcription of the SREBP-1c gene requires an endogenous sterol that activates LXRs (Laffitte et al., Proc. Natl. Acad. Sci. 98: 507-512 (2001 )).

Accordingly, compounds which function as LXR agonists would be useful in methods of increasing ABCA1 , SREBP-1c, and apoE expression, increasing HDL cholesterol and treating LXR mediated diseases or conditions such as hypercholesterolemia and cardiovascular diseases.

SUMMARY OF THE INVENTION The present invention relates to the use of compounds of the following structure (I):

I Where Y, R-i and R₂ are as defined below. Other aspects of this invention will become apparent as the description of this invention continues. Hence, the foregoing merely summarizes certain aspects of the invention and is not intended, nor should it be construed, as limiting the invention in any way.

DETAILED DESCRIPTION OF THE INVENTION

The detailed description of the invention that follows is not intended to be exhaustive or to limit the invention to the precise details disclosed. It has been chosen and described to best explain the details of the invention to others skilled in the art.

The present invention relates to the use of compounds of the following structure (I):

wherein Y is selected from S or O;

Ri and R₂ are different, and each is independently selected from hydrogen, halogen, hydroxy, nitro, cyano, C1-C12 alkyl, substituted Cι-C-|₂ alkyl, alkylamino, alkylsulfanyl, aryl, C₁-C₁2 alkoxy, substituted C-ι-Cι₂ alkoxy, -C(X)R₃, -CR₈(V)Rn, -CH₂CR₈(V)Rn, -S(0)_nR₃, -NR₄P(0)(R₅)₂, -P(0)(R₅)₂, or a substituted or unsubstituted monocyclic 3-7 membered ring containing zero to three heteroatoms, wherein such heteroatoms are selected from O, N, or S, wherein such substituents are independently chosen from Rg, or Ri or R₂ can be a group of the following formulae:

in which ring D is a substituted or unsubstituted monocyclic 3-7 membered ring containing zero to three heteroatoms, wherein such heteroatoms are selected from O, N, or S, wherein such substituents are independently chosen from Rg, and R₃ and R₄, taken together can form a substituted or unsubstituted monocyclic 3-7 membered ring containing zero to three heteroatoms, wherein such heteroatoms are selected from O, N, or S, wherein such substituents are independently chosen from Rg, or Ri and R₂ taken together can form a monocyclic 5-7 membered substituted or unsubstituted ring containing zero to three heteroatoms, wherein such heteroatoms are selected from O, N, or S, and such substituents are independently selected from Re and R₇; R₃ is selected from hydrogen, hydroxy, Cι-C-ι₂ alkyl, substituted C₁-C₁₂ alkyl, alkylamino, aryl, Ci-C-i₂ alkoxy, substituted C₁-C-₁₂ alkoxy, or a monocyclic 3-7 membered ring containing zero to three heteroatoms, wherein such heteroatoms are selected from O, N, or S, wherein such substituents are independently chosen from Rg; R is selected from hydrogen, C₁-C12 alkyl, substituted C₁-C₁₂ alkyl, aryl, or a monocyclic 3-7 membered ring containing zero to three heteroatoms, wherein such heteroatoms are selected from O, N, or S, wherein such substituents are ^• independently chosen from Rg ; R5 is selected from hydrogen, hydroxy, C1-C-₁₂ alkyl, substituted C-₁-C₁₂ alkyl, alkylamino, aryl, or a monocyclic 3-7 membered ring containing zero to three heteroatoms, wherein such heteroatoms are selected from O, N, or S, wherein such substituents are independently chosen from R_g;

Re and R are different, and independently selected from Ri and R₂, with the improviso that R₆ and R₇ can not be taken together to form a ring;

R₈ is selected from hydrogen, C-ι-Cι₂ alkyl, substituted C₁-C₁₂ alkyl, heterocyclyl, or aryl;

R₉ is selected from hydrogen, halogen, -CN -C(0)CF₃, -S(0)_nCF₃, -C(0)CH₂F, -

CH(OH)CF₃, -N(CN)₂, -C(CN)₃, -CHRι₀R₈, C C₁₂ alkyl, substituted Cι-C₁₂ alkyl, -CF₃, -(CF₂)_mCF₃, -CH(CF₃)₂, -CF(CF₃)₂, -S0₃H, alkylamino, alkylsulfanyl, aryl,

C C₁₂ alkoxy, substituted C^C^ alkoxy, -C(X)R₁₀, -CR₈(V)Rn, -CH₂CR₈(V)Rn, -

S(0)_nRιι, -S(0)₂NHMe(OH), -S(0)₂NH(2-thiazolyl), -(4-oxo-2-thioxo-thiazolidin-

5-ylidene), tetrazolyl, -CH₂(1 ,1-dioxo-1lambda*6*-thiomorpholin-4-yl), -

S(0)₂CH₂NO₂, -S(0)2CH₂S(0)₂Rιι -P(0)(OR₈)Rn, -NR₈P(0)ORn, - P(0)(NRsRn), a substituted or unsubstituted monocyclic 3-7 membered ring containing zero to three heteroatoms, wherein such heteroatoms are selected from O, N, or S;

R₁₀ is selected from C-₁-C12 alkyl, substituted d-Cι₂ alkyl, heterocyclyl, or aryl;

R₁1 is selected from C₁-C12 alkyl, substituted C₁-C₁2 alkyl, heterocyclyl, or aryl; A is a bond, -NR₄-, or -CR₄R₅-;

B is a bond, -NR₄-, -CR₄H-, -CR₄(OH)- or -CR₄R₅-;

G is -CH(CH₂)m-, >C=CH-, -N(CH₂)_m;

L is a bond, -0-, -C(O)-, -NR₅-, -CR₄H-,.CR₄(OH)-, or -CR₄R₅-, -NHNR₅-;

Q is a bond, -NR₄-, -C(O)-, -O- or -CR₄R₅-; X is O, S, NR_4> NOR₄, NCN, NN0_2l CR₈N0₂₎ CR₈CN, C(CN)_2> CR₈Rn, or N-

NR₈Rn;

V is -OH, -SH, -CN; m is zero, one, two or three; n is one or two; its corresponding enantiomers, diastereoisomers or tautomers, or a pharmaceutically acceptable salt, or a prodrug thereof in an pharmaceutically- acceptable carrier.

There are several preferred embodiments of the present invention. One such preferred embodiment is that Y is S. Another such preferred embodiment is that Ri and R₂ taken together form a substituted five to seven membered ring containing zero to three heteroatoms selected from N, O, or S, wherein such substituents are independently chosen from R₆ and R₇. Another such preferred embodiment is that Re and R are selected from -CR₈(V)Rn or - CH2CR_β(V)Rιι, wherein V is either -OH and -CN, -P(0)(OR₄)R₃₎ -P(0)(R₃)₂, or a group of the following formulae:

A particularly preferred embodiment is that the ring formed from Ri and R₂ is a 6- membered aryl containing zero to three nitrogens which is substituted according to the following formulae:

Wherein Y is S, and U is selected from C or N.

An even more preferred embodiment is that in the above described bicyclic structures, R₆ and R₇ are different, and selected from H or a group of the following formulae:

DEFINITIONS

As used herein, "alkyl" means a straight chain alkane, alkene, or alkyne substituent containing only carbon and hydrogen, such as methyl, ethyl, butyl, pentyl, heptyl and the like. Alkyl groups can be saturated or unsaturated (i.e., containing -C=C- or -Cf C- linkages), at one or several positions. When a specific degree of unsaturation is preferred, said substituent is referred to as either "alkenyl" or "alkynyl", denoting substituents containing -C=C- or -CfC- linkages, respectively. The number of carbons may be denoted as "Cj-Cj-alkyl" wherein I and j refer to the minimum and maximum number of carbon atoms, respectively. Typically, alkyl groups will comprise 1 to 12 carbon atoms, preferably 1 to 10, and more preferably 2 to 8 carbon atoms.

As used herein, "substituted alkyl" means a hydrocarbon substituent, which is linear, cyclic or branched, in which one or more hydrogen atoms are substituted by carboxy, hydroxy, alkoxy, cyano, nitro, carbonyl, aryl, carboxyalkyl, mercapto, amino, amido, ureido, carbamoyl, sulfonamido, sulfamido, or halogen. Preferred substituted alkyls have their alkyl spacers (i.e., portion which is alkyl) of 1 to about 5 carbons, and may be branched or linear, and may include cyclic substituents, either as part or all of their structure. Preferred examples of "substituted alkyls" include 4-carboxybutyl, pyridin-2-ylmethyl, and 1 ,3-thiazol-2- ylmethyl, benzyl, phenethyl, and trifluoromethyl. The term "substituted alkyl" may be combined with other art accepted terms. For example "substituted alkoxy" means alkoxy as understood in the art, wherein the alkyl portion of the substituent is substituted.

As used herein, "branched alkyl" means a subset of "alkyl", and thus is a hydrocarbon substituent, which is branched. Preferred branched alkyls are of 3 to about 12 carbons, and may include cycloalkyl within their structure. Examples of branched alkyl include isopropyl, isobutyl, 1 ,2-dimethyl-propyl, cyclopentyl methyl and the like. The term "branched alkyl" may be combined with other art accepted terms. For example "branched alkoxy" means alkoxy as understood in the art, wherein the alkyl portion of the substituent is branched.

As used herein, "cycloalkyl" is a hydrocarbon substituent that is cyclic, and can be substituted or unsubstituted. Where it is substituted, one or more hydrogen atoms are substituted by carboxy, hydroxy, alkoxy, cyano, nitro, carbonyl, aryl, carboxyalkyl, mercapto, amino, amido, ureido, carbamoyl, sulfonamido, sulfamido, or halogen. Preferred cyclic alkyls are of 3 to about 7 carbons. Examples of cycloalkyl include cyclopropyl, cyclopentyl, 4-fluoro- cyclohexyl, 2,3-dihydroxy-cyclopentyl, and the like.

As used herein, "alkylene" is an alkyl diradical, i.e., an alkyl that has open valences on two different carbon atoms. Hence "(alkylene)Rj" is an alkyl diradical attached at one carbon and having substituent R_\ attached at another carbon, which may be one or more carbons away from the point of attachment. Alkylene can be linear, branched, or cyclic. Examples of alkylene include -CH₂-, CH₂CH₂-, -(CH₂)4-, -(cyclohexyl)-, and the like.

As used herein, "aryl" is a substituted or unsubstituted aromatic, i.e., Hϋckel 4n + 2 rule applies, radical having a single-ring (e.g., phenyl) or multiple condensed rings (e.g., naphthyl or anthryl), which may contain zero to 4 heteroatoms. Hence the term "heteroaryl" is clearly contemplated in the term "aryl". Preferred carbocyclic aryl, is phenyl. Preferred monocyclic heterocycles, i.e., heteroaryls, are 5 or 6 membered rings. Preferably, where the term "aryl" represents an aromatic heterocycle, it is referred to as "heteroaryl" or "heteroaromatic", and has one or more heteroatom(s). Preferred numbers of such heteroatoms are from one to three N atoms, and preferably when "heteroaryl" is a heterocycle of five members, it has one or two heteroatoms selected from O, N, or S. Hence, preferred heterocycles have up to three, more preferably two or less, heteroatoms present in the aromatic ring. The skilled artisan will recognize that among heteroaryl, there are both five and six membered rings. Examples of "heteroaryl" include; thienyl, pyridyl, pyrimidyl, pyridazyl, furyl, oxazolyl, imidazolyl, thiazolyl, oxadiazilyl, triazinyl, triazolyl, thiadiazolyl, and others, which the skilled artisan will recognize. In this definition it is clearly contemplated that substitution on the aryl ring is within the scope of this invention. Where substitution occurs, the radical is referred to as "substituted aryl". Preferably one to three, more preferably one or two, and most preferably one substituent is attached to the aryl ring. Although many substituents will be useful, preferred substituents include those commonly found in aryl compounds, such as alkyl, hydroxy, alkoxy, cyano, nitro, halo, haloalkyl, mercapto and the like. Such substituents are prepared using known methodologies. These substituents may be attached at various positions of the aryl ring, and wherein a given placement is preferred, such placement is indicated by "o,/77,p-Rj-aryl". Thus, if substituent R is attached at the para position of the aryl, then this is indicated as "p-Rj-substituted aryl".

As used herein, "amide" includes both RNR'CO- (in the case of R = alkyl, alkaminocarbonyl-) and RCONR'- (in the case of R = alkyl, alkyl carbonylamino-). As used herein, "ester" includes both ROCO- (in the case of R = alkyl, alkoxycarbonyl-) and RCOO- (in the case of R = alkyl, alkylcarbonyloxy-).

As used herein, "halogen" is a chloro, bromo, fluoro or iodo atom radical. Chloro, bromo and fluoro are preferred halogens. The term "halogen" also contemplates terms sometimes referred to as "halo" or "halide".

As used herein, "alkylamino" is an amine radical in which at least one hydrogen atom on the nitrogen has been replaced with alkyl. Preferred examples include ethylamino, butylamino, isopropylamino, and the like. The alkyl component may be linear, branched, cyclic, substituted, saturated, or unsatu rated.

As used herein, "alkylsulfanyl" is a thiol radical in which the hydrogen atom on sulfur has been replaced with alkyl. Preferred examples include ethylsulfanyl, butylsulfanyl, isopropylsulfanyl, and the like. The alkyl component may be linear, branched, cyclic, substituted, saturated, or unsaturated.

As used herein, "alkoxy" is a hydoxyl radical in which the hydrogen atom on oxygen has been replaced with alkyl. Preferred examples include ethoxy, butoxy, benzyloxy, and the like. The alkyl component may be linear, branched, cyclic, substituted, saturated, or unsaturated.

As used herein, "heterocycle(s)" means ring systems, preferably of 3-7 members, which are saturated or unsaturated, and non-aromatic. These may be substituted or unsubstituted, and are attached to other parts of the molecule via any available valence, preferably any available carbon or nitrogen. More preferred heterocycles are of 5 or 6 members. In six-membered monocyclic heterocycles, the heteroatom(s) are from one to three of O, S, or N, and wherein when the heterocycle is five-membered, preferably it has one or two heteroatoms selected from O, N, or S.

As used herein, "heterocyclyl" means radical heterocycles. These may be substituted or unsubstituted, and are attached to other via any available valence, preferably any available carbon or nitrogen.

As used herein, "sulfamido" means an alkyl-N-S(0)₂N-, aryj-NS(0) N- or heterocyclyl-NS(0)₂N- group wherein the alkyl, aryl or heterocyclyl group is as defined herein above. As used herein, "sulfonamido" means an alkyl-S(0)₂N-, aryl-S(0)₂N- or heterocyclyl- S(0)₂N- group wherein the alkyl, aryl or heterocyclcyl group is as herein described.

As used herein, "ureido" means an alkyl-NCON-, aryl-NCON- or heterocyclyl-NCON- group wherein the alkyl, aryl or heterocyclyl group is as herein described.

A substituent referred to as a radical in this specification may form a ring with another radical as described herein. When such radicals are combined, the skilled artisan will understand that there are no unsatisfied valences in such a case, but that specific substitutions, for example a bond for a hydrogen, is made. Hence certain radicals can be described as forming rings together. The skilled artisan will recognize that such rings can and are readily formed by routine chemical reactions, and it is within the purview of the skilled artisan to both envision such rings and the methods of their formations. Preferred are rings having from 3-7 members, more preferably 5 or 6 members. Compounds described herein may have cyclic structures therein, such as a ring Ri and R₂. In that regard the skilled artisan recognizes that this method of description is routine in medicinal chemistry, though such may not rigorously reflect the chemical synthetic route. As used herein the term "ring" or "rings" when formed by the combination of two radicals refers to heterocyclic or carbocyclic radicals, and such radicals may be saturated, unsaturated, or aromatic. For example, preferred heterocyclic ring systems include heterocyclic rings, such as morpholinyl, piperdinyl, imidazolyl, pyrrolidinyl, and pyridyl. The skilled artisan will recognize that the radical of formula:

represents a number of different functionalities. Preferred functionalities represented by this structure include amides, ureas, thioureas, carbamates, esters, thioesters, amidines, ketones, oximes, nitroolefines, hydroxyguanidines and guanidines. More preferred functionalities include ureas, thioureas, amides, and carbamates.

The skilled artisan will recognize that some structures described herein may be resonance forms or tautomers of compounds that may be fairly represented by other chemical structures. The artisan recognizes that such structures are clearly contemplated within the scope of this invention, although such resonance forms or tautomers are not represented herein. For example, the structures:

clearly represent the same compound(s), and reference to either clearly contemplates the other. In addition, the compounds in this invention can be provided as prodrugs, the following of which serve as examples:

wherein R is a group (or linkage) removed by biological processes. Hence clearly contemplated in this invention are compounds provided as biohydrolyzable prodrugs, as they are understood in the art. "Prodrug", as used herein is any compound wherein when it is exposed to the biological processes in an organism, is hydrolyzed, metabolized, derivatized or the like, to yield an active substance having the desired activity. The skilled artisan will recognize that prodrugs may or may not have any activity as prodrugs. It is the intent that the prodrugs described herein have no deleterious effect on the subject to be treated when dosed in safe and effective amounts. These include for example, biohydrolyzable amides and esters. A "biohydrolyzable amide" is an amide compound which does not essentially interfere with the activity of the compound, or that is readily converted in vivo by a cell, tissue, or human, mammal, or animal subject to yield an active compound of the invention. A "biohydrolyzable ester" refers to an ester compound of the invention that does not interfere with the activity of these compounds or that is readily converted by an animal to yield an active formula (l) compound. Such biohydrolyzable prodrugs are understood by the skilled artisan and are embodied in regulatory guidelines.

Compounds and compositions herein also specifically contemplate pharmaceutically acceptable salts, whether cationic or anionic. A "pharmaceutically-acceptable salt" is an anionic salt formed at any acidic (e.g., carboxyl) group, or a cationic salt formed at any basic (e.g., amino) group. Many such salts are known in the art, as described in World Patent Publication

87/05297, Johnston et al., published September 11 , 1987 (incorporated by reference herein). Preferred counter-ions of salts formable at acidic groups can include cations of salts, such as the alkali metal salts (such as sodium and potassium), and alkaline earth metal salts (such as magnesium and calcium) and organic salts. Preferred salts formable at basic sites include anions such as the halides (such as chloride salts). Of course, the skilled artisan is aware that a great number and variation of salts may be used, and examples exist in the literature of either organic or inorganic salts useful in this manner.

Inasmuch as the compounds of the invention may contain one or more stereogenic centers, "Optical isomer", "stereoisomer", "enantiomer," "diastereomer," as referred to herein have the standard art recognized meanings (cf. Hawleys Condensed Chemical Dictionary, 1 1th Ed.) and are included in the compounds claimed, whether as racemates, or their optical isomers, stereoisomers, enantiomers, and diastereomers.

Likewise, inasmuch as the compounds of the invention may exist as "regio-isomers", specifically those compounds of formula I that have several relative orientations of the heteroatoms within ring C, reference to such isomers has the standard art recognized meaning (cf. Hawleys Condensed Chemical Dictionary, 11th Ed.) and are included in the compounds claimed.

As used herein "cardiovascular diseases" include arrhthymia, atrial fibrillation, congestive heart failure, coronary artery disease, hypertension, myocardial infarction, stroke, ventricular fibrillation, among others, particularly cardiovascular ischemia and those conditions modulated by LXR.

COMPOSITIONS The compositions of the present invention comprise:

(a) a safe and effective amount of a LXR modulating compound (I), prodrug or pharmaceutical salt thereof; and

(b) a pharmaceutically-acceptable carrier.

As discussed above, numerous diseases can be mediated by LXR related therapy. Thus, the compounds of this invention are useful in therapy with regard to conditions involving this LXR activity.

Accordingly, the compounds of this invention can therefore be formulated into pharmaceutical compositions for use in prophylaxis, management and treatment of these conditions. Standard pharmaceutical formulation techniques are used, such as those disclosed in Remington's Pharmaceutical Sciences, Mack Publishing Company, Easton, PA.

A "safe and effective amount" of a compound of the present invention is an amount that is effective, to modulate LXR activity, in a subject, a tissue, or a cell, and preferably in an animal, more preferably in a mammal, without undue adverse side effects (such as toxicity, irritation, or allergic response), commensurate with a reasonable benefit/risk ratio, when used in the manner of this invention. The specific "safe and effective amount" will, obviously, vary with such factors as the particular condition being treated, the physical condition of the patient, the duration of treatment, the nature of concurrent therapy (if any), the specific dosage form to be used, the carrier employed, the solubility of the compound therein, and the dosage regimen desired for the composition.

In addition to the subject compound, the compositions of the subject invention contain a pharmaceutically-acceptable carrier. The term "pharmaceutically-acceptable carrier", as used herein, means one or more compatible solid or liquid filler diluents or encapsulating substances which are suitable for administration to a mammal. The term "compatible", as used herein, means that the components of the composition are capable of being commingled with the subject compound, and with each other, in a manner such that there is no interaction which would substantially reduce the pharmaceutical efficacy of the composition under ordinary use situations. Pharmaceutically-acceptable carriers must, of course, be of sufficiently high purity and sufficiently low toxicity to render them suitable for administration preferably to an animal, preferably mammal being treated. Some examples of substances, which can serve as pharmaceutically- acceptable carriers or components thereof, are sugars, such as lactose, glucose and sucrose; starches, such as corn starch and potato starch; cellulose and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose, and methyl cellulose; powdered tragacanth; malt; gelatin; talc; solid lubricants, such as stearic acid and magnesium stearate; calcium sulfate; vegetable oils, such as peanut oil, cottonseed oil, sesame oil, olive oil, corn oil and oil of theobroma; polyols such as propylene glycol, glycerine, sorbitol, mannitol, and polyethylene glycol; alginic acid; emulsifiers, such as the TWEENS; wetting agents, such sodium lauryl sulfate; coloring agents; flavoring agents; tableting agents, stabilizers; antioxidants; preservatives; pyrogen-free water; isotonic saline; and phosphate buffer solutions.

The choice of a pharmaceutically-acceptable carrier to be used in conjunction with the subject compound is basically determined by the way the compound is to be administered.

If the subject compound is to be injected, the preferred pharmaceutically- acceptable carrier is sterile, physiological saline, with blood-compatible suspending agent, the pH of which has been adjusted to about 7.4. In particular, pharmaceutically-acceptable carriers for systemic administration include sugars, starches, cellulose and its derivatives, malt, gelatin, talc, calcium sulfate, vegetable oils, synthetic oils, polyols, alginic acid, phosphate buffer solutions, emulsifiers, isotonic saline, and pyrogen-free water. Preferred carriers for parenteral administration include propylene glycol, ethyl oleate, pyrrolidone, ethanol, and sesame oil. Preferably, the pharmaceutically-acceptable carrier, in compositions for parenteral administration, comprises at least about 90% by weight of the total composition.

The compositions of this invention are preferably provided in unit dosage form. As used herein, a "unit dosage form" is a composition of this invention containing an amount of a compound that is suitable for administration to an animal, preferably mammal subject, in a single dose, according to good medical practice. (The preparation of a single or unit dosage form however, does not imply that the dosage form is administered once per day or once per course of therapy. Such dosage forms are contemplated to be administered once, twice, thrice or more per day, and are expected to be given more than once during a course of therapy, though a single administration is not specifically excluded. The skilled artisan will recognize that the formulation does not specifically contemplate the entire course of therapy and such decisions are left for those skilled in the art of treatment rather than formulation.) These compositions preferably contain from about 5 mg (milligrams), more preferably from about 10 mg to about 1000 mg, more preferably to about 500 mg, most preferably to about 300 mg, of the selected compound. The compositions of this invention may be in any of a variety of forms, suitable (for example) for oral, nasal, rectal, topical (including transdermal), ocular, intracereberally, intravenous, intramuscular, or parenteral administration. (The skilled artisan will appreciate that oral and nasal compositions comprise compositions that are administered by inhalation, and made using available methodologies.) Depending upon the particular route of administration desired, a variety of pharmaceutically-acceptable carriers well-known in the art may be used. These include solid or liquid fillers, diluents, hydrotropies, surface-active agents, and encapsulating substances. Optional pharmaceutically-active materials may be included, which do not substantially interfere with the inhibitory activity of the compound. The amount of carrier employed in conjunction with the compound is sufficient to provide a practical quantity of material for administration per unit dose of the compound. Techniques and compositions for making dosage forms useful in the methods of this invention are described in the following references, all incorporated by reference herein: Modern Pharmaceutics, Chapters 9 and 10 (Banker & Rhodes, editors, 1979); Lieberman et al., Pharmaceutical Dosage Forms: Tablets (1981 ); and Ansel, Introduction to Pharmaceutical Dosage Forms 2d Edition (1976).

Various oral dosage forms can be used, including such solid forms as tablets, capsules, granules and bulk powders. These oral forms comprise a safe and effective amount, usually at least about 5%, and preferably from about 25% to about 50%, of the compound. Tablets can be compressed, tablet triturates, enteric-coated, sugar-coated, film-coated, or multiple-compressed, containing suitable binders, lubricants, diluents, disintegrating agents, coloring agents, flavoring agents, flow-inducing agents, and melting agents. Liquid oral dosage forms include aqueous solutions, emulsions, suspensions, solutions and/or suspensions reconstituted from non-effervescent granules, and effervescent preparations reconstituted from effervescent granules, containing suitable solvents, preservatives, emulsifying agents, suspending agents, diluents, sweeteners, melting agents, coloring agents and flavoring agents.

The pharmaceutically-acceptable carrier suitable for the preparation of unit dosage forms for peroral administration are well-known in the art. Tablets typically comprise conventional pharmaceutically-compatible adjuvants as inert diluents, such as calcium carbonate, sodium carbonate, mannitol, lactose and cellulose; binders such as starch, gelatin and sucrose; disintegrants such as starch, alginic acid and croscarmelose; lubricants such as magnesium stearate, stearic acid and talc. Glidants such as silicon dioxide can be used to improve flow characteristics of the powder mixture. Coloring agents, such as the FD&C dyes, can be added for appearance. Sweeteners and flavoring agents, such as aspartame, saccharin, menthol, peppermint, and fruit flavors, are useful adjuvants for chewable tablets. Capsules typically comprise one or more solid diluents disclosed above. The selection of carrier components depends on secondary considerations like taste, cost, and shelf stability, which are not critical for the purposes of the subject invention, and can be readily made by a person skilled in the art.

Peroral compositions also include liquid solutions, emulsions, suspensions, and the like. The pharmaceutically-acceptable carriers suitable for preparation of such compositions are well known in the art. Typical components of carriers for syrups, elixirs, emulsions and suspensions include ethanol, glycerol, propylene glycol, polyethylene glycol, liquid sucrose, sorbitol and water. For a suspension, typical suspending agents include methyl cellulose, sodium carboxymethyl cellulose, AVICEL RC-591 , tragacanth and sodium alginate; typical wetting agents include lecithin and polysorbate 80; and typical preservatives include methyl paraben and sodium benzoate. Peroral liquid compositions, may also contain one or more components such as sweeteners, flavoring agents and colorants disclosed above.

Such compositions may also be coated by conventional methods, typically with pH or time-dependent coatings, such that the subject compound is released in the gastrointestinal tract in the vicinity of the desired topical application, or at various times to extend the desired action. Such dosage forms typically include, but are not limited to, one or more of cellulose acetate phthalate, polyvinylacetate phthalate, hydroxypropyl methyl cellulose phthalate, ethyl cellulose, Eudragit coatings, waxes and shellac.

Compositions of the subject invention may optionally include other drug actives. Other compositions useful for attaining systemic delivery of the subject compounds include sublingual, buccal and nasal dosage forms. Such compositions typically comprise one or more of soluble filler substances such as sucrose, sorbitol and mannitol; and binders such as acacia, microcrystalline cellulose, carboxymethyl cellulose and hydroxypropyl methyl cellulose. Glidants, lubricants, sweeteners, colorants, antioxidants and flavoring agents disclosed above may also be included.

The compositions of this invention can also be administered topically to a subject, e.g., by the direct application or spreading of the composition on the epidermal or epithelial tissue of the subject, or transdermally via a "patch". Such compositions include, for example, lotions, creams, solutions, gels and solids. These topical compositions preferably comprise a safe and effective amount, usually at least about 0.1 %, and preferably from about 1 % to about 5%, of the compound. Suitable carriers for topical administration preferably remain in place on the skin as a continuous film, and resist being removed by perspiration or immersion in water. Generally, the carrier is organic in nature and capable of having dispersed or dissolved therein the compound. The carrier may include pharmaceutically-acceptable emolients, emulsifiers, thickening agents, solvents and the like.

METHODS OF ADMINISTRATION

The compounds and compositions of this invention can be administered topically or systemically. Systemic application includes any method of introducing compound into the tissues of the body, e.g., intra-articular, intrathecal, epidural, intramuscular, transdermal, intravenous, intraperitoneal, subcutaneous, sublingual administration, inhalation, rectal, or oral administration. The compounds of the present invention are preferably administered orally.

The specific dosage of the compound to be administered, as well as the duration of treatment is to be individualized by the treating clinicians. Typically, for a human adult (weighing approximately 70 kilograms), from about 5 mg, preferably from about 10 mg to about 3000 mg, more preferably to about 1000 mg, more preferably to about 300 mg, of the selected compound is administered per day. It is understood that these dosage ranges are by way of example only, and that daily administration can be adjusted depending on the factors listed above.

In all of the foregoing, of course, the compounds of the invention can be administered alone or as mixtures, and the compositions may further include additional drugs or excipients as appropriate for the indication. For example, in the treatment of cardiovascular diseases, it is clearly contemplated that the invention may be used in conjunction with beta-blockers, calcium antagonists, ACE inhibitors, diuretics, angiotensin receptor inhibitors, or known cardiovascular drugs or therapies. Hence, in this example, novel compounds or compositions of this invention are useful when dosed together with another active and can be combined in a single dosage form or composition.

The composition can also be administered in the form of liposome delivery system, such as small unilamellar vesicles, large unilamellar vesicles, and mutilamellar vesicles. Liposomes can be formed from a variety of phospholipids, such as cholesterol, stearylamine, or phosphayidylcholines.

BIOLOGICAL ACTIVITY

In Vitro LXR and β activity assay:

The assays for gene expression (apoE, ABCA1 , and SREBP-1c) are performed following the literature procedures (Sparrow et al., J. Biol. Chem. 277:10021- 10027(2002); Laffitte et al., Proc. Natl. Acad. Sci., 98:507-512(2001); Repa et al., Genes Dev. 14:2819-2830(2000)). LXR modulator refers to compounds which achieve at least 50% activation or inhibition of LXR relative to 24(S)25- epoxycholesterol, the positive control, or which stimulate the expression of the responsive genes, such as ABCA1 genes, in a cell system.

To determine the effectiveness of representative compounds of the present invention as LXR modulators, THP-1 cells were maintained in suspension for passage and growth in PRMI 1640 (Invitrogen) containing 10% FBS (Irvine Scientific, Santa Ana, CA), 100 units/ml penicillin/100ug/ml streptomycin (Irvine Scientific), 1mM sodium pyruvate (Invitrogen) and 55 uM β- mercaptoethanol (Sigma). Passaging was performed every 3-4 days at a 1 :4 dilution. For experiments, 1X106 cells/well were plated in 6-welI plates in media supplemented with 100 ng/ml phorbol 12-myristate-13-acetate (PMA, Sigma) to induce differentiation. Cells were maintained in this media for 5-days prior to treatment with LXR agonists. Generally, the culture media was replaced with media containing vehicle (DMSO or ethanol) or 1-10 uM drugs at 0 h. THP-1 cells were dosed a second time at 24 h and then harvested for RNA isolation 24 h later.

Total RNA samples were diluted to 100 ug/ml and treated with 40 units/ml RNA-free Dnase I (Ambion, Austin, TX) for 30 min at 37°C followed by inactivation at 75oC for 5 min. Samples were quantitated by spectrophotometry or with the RiboGreen assay (Molecular Probes, Eugee, OR) and diluted to a concentration of 10ng/ul. Samples were assayed in duplicate 25-ul reactions using 35ng of RNA/reaction with PerkinElmer chemistry on an ABI Prism 7700 (Applied Biosystems). Gene specific primers were used at 7.5 or 22.5 pmol/reaction and optimized for each gene examined, and the gene-specific fluorescently tagged probe was used at 5 pmol/reaction. In this system, the probe is degraded by Taq polymerase during the amplification phase, releasing the fluorescent tag from its quenched state; amplification data is expressed as the number of PCR cycles required to elevate the fluorescence signal beyond a threshold intensity level. Fold induction values were calculated by subtracting the mean threshold cycle number for each treatment group from the mean threshold cycle number of the vehicle group and raising 2 to the power of this difference.

As shown in Table A, LXR modulating activity was determined from the magnitude of gene expression induction as compared to a control (DMSO). Compounds showing significant induction of the ABCA1 gene in a THP-1 cell system, as compared to the control (DMSO), demonstrates that the compounds of the present invention are useful LXR modulators for increasing ABCA1 expression, increasing HDL cholesterol and treating LXR mediated diseases or conditions such as hypercholesterolemia and cardiovascular diseases. Table A. Gene Expression Induction.

PREPARATION OF COMPOUNDS OF THE INVENTION

The starting materials used in preparing the compounds of the invention are known, made by known methods, or are commercially available. It will be apparent to the skilled artisan that methods for preparing precursors and functionality related to the compounds claimed herein are generally described in the literature. The skilled artisan given the literature and this disclosure is well equipped to prepare any of the claimed compounds.

It is recognized that the skilled artisan in the art of organic chemistry can readily carry out manipulations without further direction, that is, it is well within the scope and practice of the skilled artisan to carry out these manipulations. These include reduction of carbonyl compounds to their corresponding alcohols, reductive alkylation of amines, oxidations, acylations, aromatic substitutions, both electrophilic and nucleophilic, etherifications, esterification, saponification and the like. These manipulations are discussed in standard texts such as March Advanced Organic Chemistry (Wiley), Carey and Sundberg, Advanced Organic Chemistry and the like.

The skilled artisan will readily appreciate that certain reactions are best carried out when other functionality is masked or protected in the molecule, thus avoiding any undesirable side reactions and/or increasing the yield of the reaction. Often the skilled artisan utilizes protecting groups to accomplish such increased yields or to avoid the undesired reactions. These reactions are found in the literature and are also well within the scope of the skilled artisan.

Examples of many of these manipulations can be found for example in T.

Greene and P. Wuts Protecting Groups in Organic Synthesis, 2^nd Ed., John

Wiley & Sons (1991). The following example schemes are provided for the guidance of the reader, and represent preferred methods for making the compounds exemplified herein. These methods are not limiting, and it will be apparent that other routes may be employed to prepare these compounds. Such methods specifically include solid phase based chemistries, including combinatorial chemistry. The skilled artisan is thoroughly equipped to prepare these compounds by those methods given the literature and this disclosure.

Scheme 1

As shown in Scheme 1 , amino-2-mercaptobenzothiazole(s) 1 , which are commercially available or easily prepared using known methods, are condensed with aldehydes or ketones and reduced with sodium cyanoborohydride to afford the corresponding /V-alkylaniline(s) 2. Compound(s) 2 are subsequently converted to the corresponding urea(s) or thiourea(s) (3, R = aliphatic, aromatic, heterocyclyl or heteroaryl), amides (4, R = aliphatic, aromatic, heterocyclyl or heteroaryl), sulfonamide(s) (5, R₄ = aliphatic, aromatic, heterocyclyl or heteroaryl), or carbamate(s) (6, R = aliphatic, aromatic, heterocyclyl or heteroaryl), under the reaction conditions depicted.

As shown in Scheme 2, thiohydantoin analogs (7) were prepared through the direct treatment of 1 with various amino acid isothiocyanato esters (R₉ = alkyl, aryl, hetrocyclyl) in hot pyridine. Scheme 2

The pyrimidine and pyridine scaffold(s) 10 are obtained as shown in Scheme 3. Iterative displacement of the chlorines in 8 (which are commercially available or prepared using known methods) by sodium diethyldithiocarbamate, and a primary amine (R₄NH₂), respectively give intermediate(s) 9, which upon reduction/thiolysis with basic sodium sulfide and condensation with potassium xanthic acid salt afford 10.

Scheme 3

The skilled artisan recognises that compounds such as 10 are used in like fashion as compound(s) 1 as described above for the preparation of the compounds in the present invention. Also understood by the skilled artisan are the use of similar pyrimidines and pyridines (i.e., regio-isomers thereof) as described herein in like fashion to 1 for the preparation of the compounds described in this invention. Other compounds described in the present invention are prepared in analogous fashion to the teachings disclosed in International Patent Application Publication No. WO 02/064136 (published August 22, 2002), International Patent Application Publication No. WO 02/058690 (published August 1 , 2002), and International Patent Application Publication No. WO 02/058698 (published August 1 , 2002), wherein the disclosures therein are incorporated herein by reference. EXAMPLES

To further illustrate this invention, the following examples are included. The examples should not be construed as specifically limiting the invention. Variations of these examples within the scope of the claims are within the purview of one skilled in the art are considered to fall within the scope of the invention as described, and claimed herein. The reader will recognize that the skilled artisan, armed with the present disclosure, and skill in the art is able to prepare and use the invention without exhaustive examples.

Trademarks used herein are examples only and reflect illustrative materials used at the time of the invention. The skilled artisan will recognize that variations in lot, manufacturing processes, and the like, are expected. Hence the examples, and the trademarks used in them are non-limiting, and they are not intended to be limiting, but are merely an illustration of how a skilled artisan may choose to perform one or more of the embodiments of the invention. ¹H nuclear magnetic resonance spectra (NMR) is measured in CDCI₃ or other solvents as indicated by a Varian NMR spectrometer (Unity Plus 400, 400 MHz for ¹H) unless otherwise indicated and peak positions are expressed in parts per million (ppm) downfield from tetramethylsilane. The peak shapes are denoted as follows, s, singlet; d, doublet; t, triplet; q, quartet; m, multiple! The following abbreviations have the indicated meanings:

Ac = acetyl

Bn = benzyl

Bz= benzoyl •

CDI = carbonyl diimidazole CH₂CI₂ = dichloromethane

DIBAL= diisobutylaluminum hydride

DMAP = 4~(dimethylamino)-pyridine

DMF= N,N-dimethylformamide

DMSO = dimethylsulfoxide ECAC = 1-[3-(dimethylamino)propyl]-3-ethylcarbodiimide hydrochloric acid

ESIMS = electron spray mass spectrometry

Et₃N = triethylamine EtOAc = ethyl acetate

HMTA = hexamethylenetetramine

LDA = lithium diisopropylamide

LHDMS = lithium bis(trimethylsilyl)amide MgS0 = magnesium sulfate

NaH = sodium hydride

NBS = N-bromosuccinimide

NCS = N-chlorosuccinimide

NH CI= ammonium chloride Ph = phenyl

Py = pyridinyl r.t.= room temperature

TFA = trifluoroacetic acid

THF = tetrahydrofuran TLC = thin layer chromatography

Tf₂0 = triflic anhydride

Alkyl group abbreviations

Me = methyl

Et = ethyl n-Pr = normal propyl i-Pr = isopropyl n-Bu = normal butyl i-Bu = isobutyl t-Bu = tertiary butyl s-Bu = seconday butyl c-Hex = cyclohexyl

Example 1. Preparation of Λ/-alkyl benzothiazoles.

Table t

Example procedure for synthesis of Λ/-alkyl benzothiazoles:

6-[(2-methylpropyl)amino]-1 ,3-benzothiazole-2-thiol (Table 1 , Example 1-1 ): Into a 500 ml round bottomed flask were added 6-amino-1 ,3-benzothiazole-2- thiol (5.0 g, 0.03 mol), hexanal (3.0 g, 0.03 mol), methanol (250 ml), glacial acetic acid (2.5 ml), water (1 ml), and NaCNBH₃ (1.9 g, 0.03 mol). The mixture was stirred for 4 h, filtered, and the filtrate was concentrated to give light yellow solid. The solid was washed with water and triturated with diethyl ether to give 4.0 g (61 %) of the title compound. ¹H NMR (DMSO-c ₆) δ = 0.88 (t, 6H), 1.79 (m, 1 H), 2.77 (t, 2H), 6.64 (d, 1 H), 6.72 (s, 1 H), 6.99 (d, 2H); ESIMS: tτ?/z 239 (M-H).

Example 2. Preparation of benzothiazole-ureido compounds.

Table 2.

Example procedure for synthesis of benzothiazole-ureido compounds:

Λ/-(2-mercapto-1 ,3-benzothiazol-6-yl)-N-(2-methylpropyl)-N'-[4-

(trifluoromethyl)phenyl]urea (Table 2, Example 2-1 ):

Into 50 ml round bottomed flask were added 6-[(2-methylpropyl)amino]-1 ,3- benzothiazole-2-thiol (52.36 mg, 0.22 mmol), dichloromethane (10 ml), and α,α,α-trifluoro-p-tolyl isocyanate (41.1 mg, 0.22 mmol). The reaction mixture was stirred 8 h, filtered, and the solid that was obtained was triturated with chloroform, followed with chloroform / methanol (9:1 mixture) to give 60 mg (64%) of the title compound. ¹H NMR (DMSO- /₆) δ = 0.84 (d, 6H), 1.66 (m, 1 H), 3.49 (d, 2H), 7.30 (s, 2H), 7.50 (d, 2H), 7.60 (d, 2H), 7.71 (s, 1H); ESIMS: m/z424 (M-H). Example 3. Preparation of benzothiazole-carbamyl compounds.

Table 3.

Example procedure for synthesis of benzothiazole-carbamyl compounds:

4-chlorophenyl 2-mercapto-1 ,3-benzothiazol-6-yl(methylpropyl)carbamate (Table 3, Example 3-1):

Into a 50 ml round bottomed flask were added 6-[(2-methylpropyl)amino]-1 ,3- benzothiazole-2-thiol (100 mg, 0.42 mmol), acetone (10 ml), N,N- diisopropylethylamine (54.2 mg, 0.42 mmol) and 4-chlorophenyl chloroformate (78.8 mg, o.42 mmol). The reaction mixture was stirred for 10 h, filtered, and the solid that was obtained was triturated with diethyl ether to afford 38 mg (48%) of the title compound. ¹H NMR (DMSO-c/₆) δ = 0.85 (d, 6H), 1.70 (m, 1 H), 3.53 (br s, 2H), 7.12 (d, 2H), 7.31 (d, 1 H), 7.41 (m, 3H), 7.79 (s, 1 H); ESIMS: m/z 393 (M+H).

Example 4. Synthesis of benzothiazole-sulfonamide compounds.

4-fluoro-N-isopropyl-N-(2-mercaptobenzothiazol-6-yl)-benzenesulfonamamide: Into a 50 ml round bottomed flask were added 6-[(2-methylpropyl)amino]-1 ,3- benzothiazole-2-thiol (200 mg, 0.89 mmol), pyridine (10 ml), and 4-fluorophenyl sulfonyl chloride (208 mg, 1.07 mmol). The reaction mixture was stirred for 1 h, then concentrated. The solid that was obtained was triturated with diethyl ether, then purified by preparative TLC (5% MeOH in CHCI₃) to afford 11 mg (3 %) of the title compound. ¹H NMR (DMSO- ₆) δ = 0.90 (d, 6H), 4.41 (m, 1 H), 6.91 (br s, 1 H), 7.24 (br s, 1H), 7.41 (m, 3H), 7.77 (m, 2H), 13.9 (br s, 1H); ESIMS: m/z 381 (M-H).

Example 5. Preparation of benzothiazole-thiohvdantoin compounds.

Table 5:

Example procedure for synthesis of benzothiazole- thiohydantoin compounds:

3-(2-mercapto-1 ,3-benzothiazol-6-yl)-2-thioxoimidazolidin-4-one (Table 5, Example 5-1 ):

Into a 13 X 100 mm screw cap Pyrex® vial were added 6-amino-1 ,3~ benzothiazole-2-thiol (108 mg, 0.59 mmol), methyl-2-isothiocyanatoacetate (94.8 mg, 0.72 mmol), and pyridine (2 ml). The vial was purged with Ar, capped, and shaken in an oven at 60 °C for 8 h. The solution was concentrated, and the residue was triturated with Et₂0 to give a rusty brown powder (59 mg, 35 %). ¹H

10 NMR (DMSO-c/e) δ = 4.27 (s, 2H), 7.26 (d, 1 H), 7.35 (d, 1 H), 7.61 (s, 1 H), 10.40 (s, 1 H), 13.86 (br s, 1 H); ESIMS: m/z 280 (M-H); mp >266 °C.

Example 6. Preparation of 5-alkylamino-thiazoir5,4-b1pyridine(or pyrimϊdine)-2- thiol(s).

Table 6.

20 Example procedure for synthesis of 5-aIkylamino-thiazol[5,4-b]pyridine-2-thiol(s):

Preparation of 5-isopropylamino-thiazol[5,4-b]pyridine-2-thiol (Table 6, Example 6-1): Step 1: Into a 1 L flask were added 2,6-dichIoro-3-nitropyridine (15.01g, 77.78 mmol) and anhydrous THF. The solution was cooled in an ice-water bath for 10 min and degassed by evacuation/purging with Ar. A solution of sodium diethyldithiocarbamate (19.34g, 85.8 mmol) in 275 mL THF was prepared and degassed, and then added dropwise over 30 min to the solution of the chloropyridine. The solution was stirred at 0 degrees for 5 h, then allowed to warm to room temperature. The solvent was evaporated, and the dark orange residue was taken up in EtoAc and washed 3x with water, then 1x with brine. The organic fraction was dried (MgS0₄), filtered, and concentrated to afford an amber oil. Purification by flash column (Si0₂ gel; 10% EtoAC/hexanes) afforded 17.3 g (73%) of an orange oil (diethyl-dithiocarbamic acid 6-chloro-3-nitro- pyridin-2-yl ester).

Step 2: Into a 250 mL round bottomed flask were added the product obtained in step 1 (12.38 g, 40.48 mmol), K₂C0₃ (5.64 g, 40.8 mmol), and acetonitrile (100 mL). Isopropylamine (3.5 mL, 40.5 mmol) was added over 5 min, and the solution was stirred overnight. The reaction solution was filtered through a 2-cm pad of Celite, concentrated, and the residue was taken up in EtOAc and washed 3x with 1 M citric acid, then brine. The organic fraction was dried (Mg S0₄), filtered, and concentrated to afford an orange-red oil. Crystallization from Et₂0/hexanes gave of 12.3 g (92%) of the title compound (9, Z=C, R₅ = isopropyl) as orange crystals. ¹H NMR (CDCI₃) δ = 1.98 (d, 6H), 1.24 (t, 3H), 1.28 (t, 3H), 3.81 (q, 2H), 4.08 (q, 2H), 5.08 (d, 1 H), 6.18 (d, 1 H), 8.19 (d, 1 H).

Step 3: Into a 250 mL round bottomed flask were added (9; Z= C, R₅ = isopropyl,

4.55 g, 13.8 mmol) and EtOH (100 mL). A solution of NaOH (5.66 g, 141 mmol) and Na₂S (5.58 g, 71.5 mmol) in 50 mL water was slowly added, and the reaction mixture was refluxed overnight under Ar. The reaction mixture was neutralized to pH 6 with citric acid, then concentrated. The residue was taken up in EtOAc, and the layers were separated. The aqueous fraction was extracted 2x with EtOAc, and the combined organic fractions were washed with brine, then dried (MgS0₄), filtered, and concentrated to afford 3-amino-6-isopropylamino- pyridine-2-thiol as a brown cake.

Step 4: Into a 250 mL round bottomed flask containing the crude product obtained in step 3 (-13.8 mmol) was added ethyl xanthic acid, potassium salt (3.42 g, 21.3 mmol), and EtOH (100 mL). The reaction mixture was refluxed 5 h under Ar, then cooled to ca. 40 degrees and decolorized with charcoal. Filtration through a 2-cm pad of Celite gave a light brown cake, which was dissolved in a minimal amount of water and acidified with acetic acid to afford the crude product as a tan solid. Filtration and washing with water followed by ether afforded 2.55 g (82% from 9) of 5-isopropylamino-thiazol[5,4-b]pyridine-2-thioI (Table 6, entry 1 ) as a tan powder. ¹H NMR (DMSO-c/₆) δ = 1.09 (d, 6H), 3.90 (m, 1H), 6.44 (d, 1H), 6.74 (d, 1H), 7.25 (d, 1 H), 13.36 (br s, 1 H); ESIMS: m/z 224 (M-H).

Example 7. Preparation of F5,4-b1pyridine-ureido compounds.

Table 7.

Example procedure for synthesis of [5,4-b]pyridine-ureido compounds:

3-(4-butoxy-phenyl)-1-ethyl-1-(2-mercapto-thiazolo[5,4-b]pyridin-5-yl)-urea (Table 7, Example 7-1 ):

Into 13x100 mm pyrex reaction vial were added 5-ethylamino-thiazolo[5,4- b]pyridine-2-thioI (105 mg, 0.49 mmol), anhydrous pyridine (2 ml), and 4- butoxyphenyl isocyanate (135 μl, 0.74 mmol). The reaction mixture was stirred 3.5 h, then concentrated. Purification by preparative TLC (40% EtoAc in Hexanes) gave 26 mg (13%) of the title compound. ¹H NMR (DMSO-c₆) δ = 0.91 (t, 3H), 1.10 (t, 3H), 1.39 (m, 2H), 1.61 (m, 2H), 3.91 (m, 1 H), 6.80 (d, 2H), 7.30 (m, 3H), 7.99 (br d, 1 H), 9.43 (s, 1 H); ESIMS: m/z 401 (M-H); mp 134.3 - 135 C.

Example 8. Preparation of f5,4-blpyridine-amido compounds.

Table 8.

Example procedure for synthesis of [5,4-b]pyridine-amido compounds:

N-ethyl-N-(2-mercapto-thiazolo[5,4-b]pyridin-5-yl)-2-phenoxy-acetamide (Table 8, Example 8-1):

Into 13x100 mm pyrex reaction vial were added 5~ethylamino~thiazolo[5,4- b]pyridine-2-thiol (99 mg, 0.47 mmol), anhydrous pyridine (1.5 ml), and 4- butoxyphenyl isocyanate (100 μl, 0.72 mmol). The reaction mixture was stirred 2.5 h, then concentrated. Purification by preparative TLC (40% EtoAc in Hexanes) gave 36 mg (22%) of the title compound. ¹H NMR (DMSO- /₆) δ = 1.02 (t, 3H), 3.73 (m, 2H), 6.70 (d, 2H), 6.86 (t, 1 H), 7.19 (t, 2H), 7.29 (d, 1 H), 7.59 (br d, 1 H; ESIMS: m/z 344 (M-H); mp 107.5- 109.0 C.

Example 9. Synthesis of [5,4-blpyridine-carbamyl compounds.

Into 1.5 ml microwave reaction vial were added 5-isopropylamino-thiazolo[5,4- b]pyridine-2-thiol (50 mg, 0.22 mmol), anhydrous pyridine (1.5 ml), and 4- chlorophenyl chloroformate (40 μl, 0.29 mmol). The reaction mixture was heated by microwave at 120 C for 15 min, then concentrated. Purification by preparative TLC (40% EtoAc in Hexanes) gave 10.2 mg (12%) of the title compound. ¹H NMR (DMSO-c/e) δ = 1.21 (d, 6H), 4.42 (m, 1 H), 7.17 (d, 2H), 7.39 (d, 2H), 7.70 (d, 1 H), 8.39 (d, 1H; ESIMS: m/z 378 (M-H).

Claims

We claim:

1. A method for modulating Liver X Receptor (LXR) which comprises the administration of a therapeutically effective amount of a member selected from the group consisting of compounds of the following formula (I):

wherein Y is selected from S or O;

Ri and R₂ are different, and each is independently selected from hydrogen, halogen, hydroxy, nitro, cyano, C₁-C₁2 alkyl, substituted Cι-C-ι₂ alkyl, alkylamino, alkylsulfanyl, aryl, C₁-C₁₂ alkoxy, substituted Cι-Cι₂ alkoxy, - C(X)R_3l -CR₈(V)R₁₁, -CH₂CR₈(V)R₁₁, -S(0)_nR₃, -NR₄P(0)(R₅)₂, -

P(0)(R₅)₂, or a substituted or unsubstituted monocyclic 3-7 membered ring containing zero to three heteroatoms, wherein such heteroatoms are selected from 0, N, or S, wherein such substituents are independently chosen from Rg, or Ri or R₂ can be a group of the following formulae:

in which ring D is a substituted or unsubstituted monocyclic 3-7 membered ring containing zero to three heteroatoms, wherein such heteroatoms are selected from O, N, or S, wherein such substituents are independently chosen from Rg, and R₃ and R₄, taken together can form a substituted or unsubstituted monocyclic 3-7 membered ring containing zero to three heteroatoms, wherein such heteroatoms are selected from O, N, or S, wherein such substituents are independently chosen from Rg, or R-i and R₂ taken together can form a monocyclic 5-7 membered substituted or unsubstituted ring containing zero to three heteroatoms, wherein such heteroatoms are selected from O, N, or S, and such substituents are independently selected from Re and R₇; R₃ is selected from hydrogen, hydroxy, C-₁-C₁₂ alkyl, substituted C-₁-C₁₂ alkyl, alkylamino, aryl, C1-C12 alkoxy, substituted Cι-C₁₂ alkoxy, or a monocyclic 3-7 membered ring containing zero to three heteroatoms, wherein such heteroatoms are selected from O, N, or S, wherein such substituents are independently chosen from R_g;

R₄ is selected from hydrogen, Cι-C₁₂ alkyl, substituted C₁-C₁₂ alkyl, aryl, or a monocyclic 3-7 membered ring containing zero to three heteroatoms, wherein such heteroatoms are selected from O, N, or S, wherein such substituents are independently chosen from Rg ; R₅ is selected from hydrogen, hydroxy, C-ι~Cι₂ alkyl, substituted Cι-Cι₂ alkyl, alkylamino, aryl, or a monocyclic 3-7 membered ring containing zero to three heteroatoms, wherein such heteroatoms are selected from O, N, or S, wherein such substituents are independently chosen from Rg;

Re and R₇ are different, and independently selected from Ri and R₂, with the improviso that R₆ and R are not taken together to form a ring;

R₈ is selected from hydrogen, C-ι-Cι₂ alkyl, substituted C₁-C₁₂ alkyl, heterocyclyl, or aryl;

R₉ is selected from hydrogen, halogen, -CN -C(0)CF₃, -S(0)_nCF₃, -C(0)CH₂F, - CH(OH)CF₃, -N(CN)₂, -C(CN)₃, -CHRι₀R₈, C-rds alkyl, substituted d- C₁₂ alkyl, -CF₃, -(CF₂)mCF₃, -CH(CF₃)₂, -CF(CF₃)₂, -S0₃H, alkylamino, alkylsulfanyl, aryl, C1-C₁2 alkoxy, substituted C1-C12 alkoxy, -C(X)R₁₀, - CR₈(V)R_{11 ;} -CH₂CR₈(V)Rn, -S(0)_nRn, -S(0)₂NHMe(OH), -S(0)₂NH(2- thiazolyl), -(4-oxo-2-thioxo-thiazolidin-5-ylidene), tetrazolyl, -CH₂(1 ,1- dioxo-1lambda*6*-thiomorpholin-4-yl), -S(0)₂CH₂N0₂, -S(0)₂CH₂S(0)₂Rn -P(0)(OR₈)Rn, -NR₈P(0)ORn, -P(0)(NR₈Rn), a substituted or unsubstituted monocyclic 3-7 membered ring containing zero to three heteroatoms, wherein such heteroatoms are selected from O, N, or S;

R-io is selected from C1-C12 alkyl, substituted C1-C₁2 alkyl, heterocyclyl, or aryl;

R₁₁ is selected from C-ι-Cι₂ alkyl, substituted C₁-C₁₂ alkyl, heterocyclyl, or aryl; A is a bond, -NR4-, or -CR₄R₅-;

B is a bond, -NR₄-, -CR4H- -CR₄(OH)- or -CR₄R₅-;

G is -CH(CH₂)_m-, >C=CH-, -N(CH₂)_m;

L is a bond, -0-, -C(O)-, -NR₅-, -CR H- _CR₄(OH)-, or -CR₄R₅-, -NHNR5-; Q is a bond, -NR₄-, -C(O)-, -O- or -CR₄R₅-;

X is O, S, NR₄, NOR₄, NCN, NN0₂, CR₈N0₂, CR₈CN, C(CN)₂, CR₈Rn, or N- NR₈Rιι;

V is -OH, -SH, -CN; m is zero, one, two or three; n is one or two; its corresponding enantiomers, diastereoisomers or tautomers, or a pharmaceutically acceptable salt, or a prodrug thereof in an pharmaceutically- acceptable carrier.

2. A method according to Claim 1 wherein Y is S.

3. A method according to Claim 2 wherein Ri and R₂ taken together form a substituted 5 or 6-membered aryl containing zero to three heteroatoms selected from N, O, or S, wherein such substituents are selected from R₆ and R₇.

4. A method according to Claim 3, in which I is selected from the following formulae:

wherein U is selected from C or N, and R₆ and R₇ are defined as above.

A method according to Claim 4 wherein I has the formula:

wherein U is selected from C or N, R₇ is H, and R₆ is defined as above.

6. A method according to Claim 5 wherein R₆ is chosen from the following groups:

wherein X, R₄, and R₅ are defined as above.

7. A method according to Claim 6 wherein U is C and R_δ is selected from the following groups:

wherein R₄ and R5 are as defined above.

8 A method according to claim 1 comprising the administration of a composition containing a compound selected from the group consisting of:

Isopropyl-(2-mercapto-benzothiazol-6-yl)-carbamic acid 4-chlorophenyl ester,

3-(4-Cyano-phenyl)-1 -isopropyl-1 -(2-mercapto-thiazolo[5,4-b]pyridin-5-yl)-urea, 3-(4-Cyano-phenyl)-1 -isopropyl-1 -(2-mercapto-benzothiazol-6-yl)-urea, N-Ethyl-N-(2-mercapto-thiazolo[5,4-b]pyridin-5-yl)-4-trifluoromethyl-benzamide, 1-lsopropyl-1-(2-mercapto-benzothiazol-6-yl)-3-(4-trifuoromethyl-phenyl)-urea, lsopropyl-(2-mercapto-benzothiazo[5,4-b]pyridin-5-yl)-carbamic acid 4- chlorophenyl ester.