TRICYCLIC COMPOUNDS USEFUL FOR MODULATING LXR
FIELD OF THE INVENTION
The current invention relates to the fields of medicinal organic chemistry, pharmacology, and medicine. Further, the current invention relates to a group of compounds that demonstrate utility for treating pathological states of dislipidemia and/or atherosclerosis through modulation of liver X receptor (s) .
BACKGROUND OF THE INVENTION
Dislipidemia and its pathological sequelae, e.g., atherosclerosis, elevated blood pressure, stroke, etc., are a major cause of death, morbidity, and economic loss in the human population. The lowering of plasma lipids, especially cholesterol by various pharmacologic interventions as well as changes in life-style behavior has had a great impact on the improvement of cardio-vascular health of people throughout the world. Especially important has been the introduction and wide spread use of cholesterol lowering agents such as the chemical class of compounds known as the statins, e.g., lovostatin, etc. (For further information see: Goodman and Gilman's, 'The Pharmacological Basis of Therapeutics', 8th Ed., 1990, McGraw-Hill, Sec. VII, pp. 874-896.) Although these compounds have been very effective in improving patients with hyperlipidemia, especially hypercholesterolemia, there are still areas for further beneficial interventions. Compounds of the statin class have been found to lower the plasma lipid component known as LDL (low-density lipoprotein) or 'bad cholesterol'; however, they have little or no effect on HDL (high-density
lipoprotein) or 'good cholesterol'. The elevation of plasma HDL levels has been correlated with protection of coronary pathology independent of changes in LDL levels. Further, patients suffering from genetic pre-dispositions of HDL deficiency such as Tangier's disease, suffer from early and often fatal cardio-vascular pathology linked to dislipidemic sequelae. Thus, it would be an improvement in the art if therapeutic agents that increased HDL were available.
The plasma levels of HDL are regulated by a complex series of biological actions, which synthesize and degrade various components of the HDL particle. Regulation of these biologic pathways occurs at both the enzymatic level and at the gene regulation level. (For further information, see: Repa, J.J., et al . , 'Regulation of Absorption and ABC1- Mediated Efflux of Cholesterol by RXR Heterodimers" , Science, 289, pp. 1524-1529 (2000), Spencer, T.A., et al . ,
' Pharmacophore Analysis of the Oxysterol Receptor LXRα' , J. Med. Chem. , 2001, 44, pp. 886-897, Ou, J., et al . , 'Unsaturated fatty acids inhibit transcription of the sterol regulatory element-binding protein-lc (SREBP-lc) gene by antagonizing ligand-dependent activation of the LXR', PNAS, 98, 11, pp. 6027-6032 (2001), and references cited therein.)
The nuclear hormone receptor LXR and its ensuing gene activation is a major regulatory pathway for the metabolic fate of plasma HDL and metabolism of cholesterol. Gene products under the influence of LXR regulation include key enzymes such as ABC1 and CYP7A1 inter alia .
Recently, compounds have been reported in the literature which demonstrate the ability to up-regulate and activate (agonize) the effect of LXR, thus increasing HDL. (See: Li et al . , LXR Modulators, WO 00/54759 (2000).) Although these compounds have demonstrated beneficial effects on plasma HDL levels, it has also been reported that
they increase serum triglyceride levels which is an undesirable side-effect.
It would be useful if novel agents were available which demonstrated an enhanced and beneficial profile of lipid regulation by modulating the nuclear hormone receptor, LXR.
SUMMARY OF THE INVENTION
The current invention provides novel compounds of formula I :
Rl and Rη are independently selected from -H, -
OH, or -ORa, wherein Ra is a hydroxyl protecting group or -
CORb, or Ri and R^ ' taken together form a keto f nction;
R2 and R2 ' are independently selected from the group consisting of -H, -C_.-C6 alkyl, phenyl, or substituted phenyl, -OH, -0Ra, or R2 and R2 ' taken together to form a keto function;
R3 and R3 ' are independently selected from the group consisting of -H, -C_,-C6 alkyl, phenyl, or substituted phenyl, -OH, -0Ra; or R3 and R3 ' taken together form a keto function;
Rj-) is selected from the group consisting of -C_.-C6 alkyl, -C3-C7 cycloalkyl, phenyl, aryl , alkylaryl , and alkylheterocyclic;
R4 is selected from the group consisting of -H,
-OH, -ORa, -C_,-C6 alkyl, phenyl, or substituted phenyl; R4' is -H;
R5 is a group selected from hydrogen, -C_.-C6 alkyl, phenyl, or substituted phenyl;
Rς, and R7 are each independently selected from the group consisting of -H, -Cι-C6 alkyl, -C2-C8 alkenyl , phenyl, or substituted phenyl;
Rg and R9 are each independently selected from -H,
-Cι-C6 alkyl, phenyl, or substituted phenyl, -halo, -N02, and -NR12R13, -CONR14R15, and -COORig.
R10 is _H' 0H' 0Ra/ CORa, -Cι-C6 alkyl, -C2-C6 alkenyl, -C2-C6 alkynyl, phenyl, or substituted phenyl, - CH2ORa, -CHO, -CONRι4R15, or -COOR16;
R]_. is ~H# -Cι-C6 alkyl, -C2-C8 alkenyl, phenyl or substituted phenyl, aryl , alkylaryl, or alkylheterocycle;
R]_2 an<3 χ3 are independently selected from -H, - C_.-C6 alkyl, -C3-C7 cycloalkyl, phenyl, aryl, alkylaryl, or R^2 taken together with R]_3 forms a 4, 5, 6, or 7-membered heterocyclic ring containing a nitrogen atom;
R_4 and R15 are each independently selected from -
H, -Cι-C6 alkyl, -C3-C7 cycloalkyl, phenyl, aryl, alkylaryl, or taken together form a 4, 5, 6, or 7-membered heterocyclic ring containing a nitrogen atom;
Rig is -H, -C_.-C6 alkyl, phenyl, substituted phenyl , or benzyl ; provided that when R9 is pyrolidine, R5 is methyl, and R^Q is carboxyethyl ester group, and R^ is in a trans relationship to R5, then R]_ is not -OH; and also provided that if Ri and Ri' are -OH and H respectively, or taken together to form a ketone, then R9 can not be pyrolidinyl and Rτ_o can not be methyl, or hydroxylmethyl ;
or a pharmaceutically acceptable salt or solvate thereof.
Contemplated within the scope of the current invention are the geometric isomers associated with the asymmetric carbon atoms in compounds of formula I .
The current invention also provides methods for modulating LXR comprising the use of a compound of formula I:
wherein:
Rl and R]_' are independently selected from -H, -
OH, or -ORa, wherein Ra is a hydroxyl protecting group or -
COR]-,, or Ri and R^' taken together form a keto function;
R2 and R2 ' are independently selected from the group consisting of -H, -C_.-C6 alkyl, phenyl, or substituted phenyl, -OH, -ORa, or R2 and R2 ' taken together to form a keto function;
R3 and R3' are independently selected from the group consisting of -H, -C_.-C6 alkyl, phenyl, or substituted phenyl, -OH, -ORa; or R3 and R3 ' taken together form a keto function;
Rk is selected from the group consisting of -C_.-C6 alkyl, -C3-C7 cycloalkyl, phenyl, aryl, alkylaryl, and alkylheterocyclic ;
R4 is selected from the group consisting of -H,
-OH, -ORa, -Cι-C6 alkyl, phenyl, or substituted phenyl; R ' is -H;
R5 is a group selected from hydrogen, -Cι-C3 alkyl, phenyl, or substituted phenyl;
Rg, and R7 are each independently selected from the group consisting of -H, -C_.-C6 alkyl, -C2-C8 alkenyl, phenyl, or substituted phenyl;
Rg and R9 are each independently selected from -H,
-C_.-C6 alkyl, phenyl, or substituted phenyl, -halo, -N02, and -NR12Ri3, -CONR14R15, and -COORιS .
R10 is _H' 0H' 0R / CORa, -Ci-Ce alkyl, -C2-C6 alkenyl, -C2-C6 alkynyl , phenyl, or substituted phenyl, - CH2ORa, -CHO, -CONR14R15, or -COOR16;
Rll is -H, -Cι-C6 alkyl, -C2-C8 alkenyl, phenyl or substituted phenyl, aryl, alkylaryl, or alkylheterocycle;
R12 an Rχ3 are independently selected from -H, - Cι-C6 alkyl, -C3-C7 cycloalkyl, phe nyl , aryl, alkylaryl, or Rl2 taken together with R13 forms a 4, 5, 6, or 7-membered heterocyclic ring containing a nitrogen atom;
R^4 and R15 are each independently selected from -
H, -C_.-C6 alkyl, -C3-C7 cycloalkyl, phe nyl, aryl, alkylaryl, or taken together form a 4, 5, 6, or 7-membered heterocyclic ring containing a nitrogen atom;
Rig is -H, -Cι-C6 alkyl, phenyl, substituted phenyl , or benzyl ; or a pharmaceutically acceptable salt or solvate thereof.
The present invention also provides a compound of formula I
I wherein :
R^ and R^ ' are independently selected from -H, -
OH, or -ORa, wherein Ra is a hydroxyl protecting group or - COR]-,, or Ri and Rχ; taken together form a keto function; R2 and R2 ' are independently selected from the group consisting of -H, -C_.-C6 alkyl, phenyl, or substituted phenyl, -OH, -ORa, or R2 and R2 ' taken together to form a keto function;
R3 and R3 ' are independently selected from the group consisting of -H, -Cι-C6 alkyl, phenyl, or substituted phenyl, -OH, -ORa; or R3 and R3 ' taken together form a keto function;
R]-, is selected from the group consisting of -C_.-C6 alkyl, -C3-C7 cycloalkyl, phenyl, aryl, alkylaryl, and alkylheterocyclic;
R4 is selected from the group consisting of -H,
-OH, -0Ra, -Cι-C6 alkyl, phenyl, or substituted phenyl; R4' is -H;
R5 is a group selected from hydrogen, -C_.-C3 alkyl, phenyl, or substituted phenyl;
Rg , and R7 are each independently selected from the group consisting of -H, -C;_-C6 alkyl, -C2-C8 alkenyl, phenyl, or substituted phenyl;
Rg and R9 are each independently selected from -H, -C_.-C6 alkyl, phenyl, or substituted phenyl, -halo, -N02, and -NR12R13, -CONR14R15, and -COORι6.
R10 is -H, OH, ORa, CORa, -Cι-C6 alkyl, -C2-C6 alkenyl, -C2-C6 alkynyl, phenyl, or substituted phenyl, - CH2ORa, -CHO, -CONR14R15 , or -COOR16;
R^. is -H' -Cι-C6 alkyl, -C2-C8 alkenyl, phenyl or substituted phenyl, aryl, alkylaryl, or alkylheterocycle;
R^_2 an( R-13 a*"e independently selected from -H, - Cι-C6 alkyl, -C3-C7 cycloalkyl, phenyl, aryl, alkylaryl, or R12 taken together with R^3 forms a 4, 5, 6, or 7-membered heterocyclic ring containing a nitrogen atom;
Rl4 and R15 are each independently selected from -
H, -Cι-C6 alkyl, -C3-C7 cycloalkyl, phenyl, aryl, alkylaryl, or taken together form a 4, 5, 6, or 7-membered heterocyclic ring containing a nitrogen atom;
Rl6 is -H, -Cχ-C6 alkyl, phenyl, substituted phenyl , or benzyl ; provided that when R9 is pyrolidine, R5 is methyl, and R^Q is carboxyethyl ester group, and R^ is in a trans relationship to R5 , then R^ is not -OH; and also provided that if Ri and Ri' are -OH and H respectively, or taken together to form a ketone, then R9 can not be pyrolidinyl and Rio can not be methyl, or hydroxylmethyl ; or a pharmaceutically acceptable salt or solvate thereof, useful in the manufacture of a medicament for the treatment and/or prevention of LXR mediated diseases diseases including dislipidemia and /or atherosclerosis.
The geometric isomers associated with the asymmetric carbon atoms of compounds of formula I are also contemplated
to be within the scope of the current invention as useful for the treatment of diseases related to LXR modulation.
A further aspect of the current invention provides methods of treatment and/or prevention of the harmful effects due to dislipidemia and some other risk factors leading to atherosclerosis. Such methods comprise the administration of an effect dose of a compound of formula I in a pharmaceutically acceptable formulation to a patient in need thereof .
The present invention also provides compounds useful for the activation of LXR transcriptional activity.
The present invention also provides compounds useful as regulators of lipid levels.
The present invention also provides a combination therapy involving a compound of formula I and other cardiovascular agents such as for example statins, leptin, and/or RXR regulating agents useful for the treatment and/or prevention of atherosclerosis.
DETAILED DESCRIPTION OF THE INVENTION
The current invention provides for novel compounds of formula I and compounds of formula I useful in modulating LXR.
The term "modulation" would include, but not be limited to, up-regulation, down-regulation, inhibition, agonism, antagonism of the LXR receptor and the resulting biological sequelae from such intervention. Also, this concept of modulation includes the selective intervention of
various sub-types of the LXR receptor family, e.g., LXR α or β, etc. The phrase "diseases" or "diseases related to LXR modulation" refers to pathological states where atherosclerosis and cardiovascular diseases are prone because of dislipidemia and other risk factors and are therefore beneficially affected by up-regulation (modulation) of LXR. These diseases include but are not limited to hyperlipidemia and it sequelae such as atherosclerosis, elevated blood pressure, stroke, and hypertriglyceremia .
The term "treatment" bears its usual meaning which includes prohibiting, inhibiting, ameliorating, halting, restraining, slowing or reversing the progression, or reducing the severity of a pathological symptom related to or resultant from the modulation of LXR, especially as related to raising plasma levels of HDL, controlling atherosclerosis, hyperlipidemia and/or hypercholesterolemia.
General chemical terms used in the description of compounds herein described bear their usual meanings. For example, the term "Ci-6 alkyl," or " (Cι-C6) alkyl" or "C_-C6 alkyl refers to straight or branched aliphatic chains of 1 to 6 carbon atoms including but not limited to methyl, ethyl, propyl , iso-propyl, n-butyl, pentyl , and hexyl . The term alkylaryl refers to an alkyl group i.e. C_,-C6 alkyl attached to an aryl group and whereby the resulting group (alkylaryl group) is attached to the nucleus via the alkyl group .
The term "substituted phenyl" refers to a phenyl group having one or more substituents selected from the group consisting of Cι-C6 alkyl, Cι-C6 alkoxy, hydroxy, nitro, chloro, fluoro, bromo, iodo, or tri (chloro or fluoro) methyl .
The term, "pharmaceutically accepted salt", refers to any one of the base addition salts which are known to be
non-toxic and are commonly used in the pharmaceutical literature. Commonly used basic addition salts would be the salts formed by: alkali or alkaline earth hydroxides, ammonium hydroxide, sodium hydroxide, alkyl or aromatic amines and the like.
The term "aryl" refers to a substituted or unsubstituted aromatic or heteroaromatic radical (wherein the terms "aromatic group" and "heteroaromatic group" refer to common aromatic rings having 4n + 2 pi electrons in a monocyclic or bicyclic conjugated system) selected from the group consisting of 2-furyl, 3-furyl, 2-thienyl 3- thienyl, 1-pyrrolyl, 2-pyrrolyl, 3-pyrrolyl, phenyl, benzyl, 2- pyridyl, 3-pyridyl, 4-pyridyl, 1-naphthyl, 2-naphthyl, 2- benzofuryl, 3-benzofuryl, 4-benzofuryl , 5-benzofuryl , 6- benzofuryl, 7-benzofuryl , 2-benzothieny, 3-benzothienyl , 4- benzothienyl , 5-benzothienyl , 6-benzothienyl , 7- benzothienyl , 1-indolyl, 2-indolyl, 3-indolyl, 4-indolyl, 5- indolyl, 6-indolyl and 7-indolyl. Aryl groups may be optionally substituted at one or two carbon atoms of the aryl group, and may be with C1-4 alkyl, C1-4 alkoxy, halogen, -NO2 , -CN, -COOH, -CONH2, -SO3H, -SO2NH2 or trifluoromethyl . Examples of substituted aryl groups are 4- methyl-3-furyl , 3 , 4 -dimethyl -2 -thienyl , 2 , 4-dimethyl-3- thienyl, 3-ethoxy-4-methyl-2-benzofuryl , 2-cyano-3- benzofuryl, 4-trifluoromethyl-2 -benzothienyl , and the like.
The term "solvate" represents an aggregate that comprises one or more molecules of the solute, such as a formula I compound, with one or more molecules of solvent.
The term, "hydroxy protecting group" (Ra) contemplates numerous functionalities used in the literature to protect a hydroxyl function during a chemical sequence and which can be removed to yield the alcohol . Included within this group would be acyls, mesylates, tosylates, benzyl, -0(C]_-C3) alkyls, and the like. Numerous reactions
for the formation and removal of such protecting groups are described in a number of standard works including, for example, Protective Groups in Organic Chemistry, Plenum Press (London and New York, 1973); Green, T.W., Protective Groups in Organic Synthesis, Wiley, (New York, 1981) ; and The Peptides, Vol. I, Schrooder and Lubke, Academic Press (London and New York, 1965) .
The term "heterocycle" or "heterocyclic" refers to a 5, 6 or 7 membered saturated, partially unsaturated, or aromatic mono-cyclic or benzofused bicyclic ring containing 1-5 heteroatoms selected from N, S or O, wherein said heterocycle is optionally substituted 1-4 times with C_.-C6 alkyl groups. Most preferred hereocyclic groups include pyrolidinyl, piperidinyl , hexamethyleneimmino, morpholino, and the like. As a corollary, the term "alkylheterocyclic" or "alkylheterocycle" is understood to mean that the alkyl group is attached to the heterocycle and that the point of attachment to the molecular backbone or nucleus is the alkyl group .
The term "molecular backbone" or "nucleus" refers to the tricyclic nucleus (X) or core of the molecule i.e. the molecule depicted by formula I without substitutents as shown below:
(X) The term "carbocycle" or carbocyclic" as used herein refers to a 3, 4, 5, 6, 7, 8, 9, or 10 members ring consisting entirely of carbon atoms and may be saturated,
fully or partially unsaturated, aromatic or non aromatic and may be optionally substituted 1-4 times with C!-C6 alkyl groups .
The term "Prodrugs" describes derivatives of the compounds of the invention which have chemically or metabolically cleavable groups and become by solvolysis or under physiological conditions the compounds of the invention which are pharmaceutically active in vivo. Derivatives of the compounds of this invention have activity in both their acid and base derivative forms, but the acid derivative form often offers advantages of solubility, tissue compatibility, or delayed release in a mammalian organism (see, Bundgard, H. , Design of Prodrugs, pp. 7-9, 21-24, Elsevier, Amsterdam 1985) . Prodrugs include acid derivatives, such as, esters prepared by reaction of the parent acidic compound with a suitable alcohol, or amides prepared by reaction of the parent acid compound with a suitable amine . Simple aliphatic esters (e.g., methyl, ethyl, propyl, isopropyl, butyl, sec-butyl, tert-butyl) or aromatic esters derived from acidic groups pendent on the compounds of this invention are preferred prodrugs. Other preferred esters include morpholinoethyloxy, diethylglycolamide and diethylaminocarbonylmethoxy. In some cases it is desirable to prepare double ester type prodrugs such as (acyloxy) alkyl esters or ( (alkoxycarbonyl) oxy) alkyl esters .
Preferred Embodiments of The Invention
Preferred R]_ and R]_ '
Preferred compounds of the present invention include compounds of formula I wherein R^ is selected from -OH,
-OCH3, -OC2H5 and Ri' is hydrogen. Most preferred are compounds of formula I wherein R^ is OH, R]_' is H, and R^ is in a trans relationship with R5.
Preferred R2 and R2 '
Preferred compounds of the present invention include compounds of formula I wherein R2 is selected from H, -OH,
-OCH3, -OC2H5, Cι-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, phenyl, benzyl, and R2 ' is hydrogen. Most preferred are compounds of formula I wherein R2 and R2 ' are both hydrogen atoms .
Preferred R3 and R3 '
Preferred compounds of the present invention include compounds of formula I wherein R3 is selected from -H, -OH,
OCH3, -OC2H5, -C_,-C6 alkyl, -C2-C6 alkenyl, -C2-C3 alkynyl, phenyl, benzyl, and R3 ' is hydrogen. Most preferred are compounds of formula I wherein R3 and R3 ' are both H.
Preferred R4 and R4 '
Preferred compounds of the present invention include compounds of formula I wherein R4 is selected from H, OH,
OCH3, -OC H5, -Cι-C6 alkyl, -C2-C6 alkenyl, -C2-C6 alkynyl, phenyl, benzyl, and wherein R4 ' is hydrogen. Most preferred are compounds of formula I wherein R4 and R4 ' are both hydrogen atoms .
Preferred R5
Preferred R5 groups include hydrogen, C_-C6 alkyl. Most preferred R5 groups include hydrogen and C^-C6 alkyl in a cis relationship at the bridgehead hydrogen and in a trans relationship with R^ .
Preferred Rg, and R7
Preferred Rg, and R7 are each independently selected from the groups represented by is -H, -C_.-C6 alkyl, -C2-C6 alkenyl, -C2-C6 alkynyl, phenyl, and benzyl.
Preferred Rg and R9
A preferred embodiment of the current invention include compounds of formula I in which Rg and Rg are indedpendently selected from pyrolidinyl or piperidinyl or morpholino.
Preferred R^Q
Yet another preferred embodiment include compounds of formula I wherein R10 is methyl, -COOMe, COOEt or -COOiPr.
Preferred R^
A preferred embodiment of the invention consist of compounds of formula I wherein Rn is selected from ~H. ~Cι- C6 alkyl and benzyl .
Also preferred is an embodiment of the invention wherein the angular methyl and adjacent hydrogen at the decalin bridgehead is in the cis configuration.
A most preferred embodiment of the current invention is the use of a compound of formula I wherein Rl', and R2 ' are hydrogen, R^ is hydroxy, R^Q i-s -COOEt, Rg
is pyrolidinyl, the angular methyl and adjacent hydrogen at the decalin bridgehead is in a cis configuration, and the R] is in a trans- configuration relative to the angular hydrogen .
The compound of formula III was commercially available from Aldrich Chemical Co., Milwaukee WI , USA.
III
However, until the present invention there is no known utility for the compound of formula III, and the compounds of formula I .
Certain compounds of formula I may be synthesized from the compound of formula III by chemical transformations well known in the art. Compounds of formula I wherein R3 is
-COOH, -COORb, other than -COOEt, or -CONR8R9 may be prepared by hydrolysis of the ethyl ester to the carboxylic acid and subsequent conversion to other esters and amides by methods known to one of skill in the art i.e. use of coupling agent such as EDCI or DCU (ethylenediamine carbodiimide or dicyclohexylurea) for making amides or reaction of the resulting free acid with ammonia to substited amines to form primary or secondary amides .
Compounds where Rio is methyl, hydroxylmethyl, or an aldehyde may be prepared by reduction of the precursor carboxylic acid by reducing agents well known in the art.
Making Compounds of the Invention
Synthesis of representative compounds of the present invention is shown in Scheme I below:
Scheme 1
Preparation of compound 3 is described in Ruel et al . Org. Synthesis 1997, 75, 69. Compound 1 is readily available from commmercial chemical sources such as for example Aldrich Chemical Company, Milwaukee, USA. Halogenation of 1 is usually performed using iodine or bromine in pyridine or a mixture of pyridine and an ethereal solvent such as for example diethyl ether. Variously substituted analogs of a compound of formula I may be. made by starting with appropriately substituted analogs of compound 1. Boronic acid coupling of 2 gives ketone 3 which can be treated with an organometallic reagent such as vinylmagnesium bromide or substituted vinylmagnesium bromide to give compound 4 or substituted analogs thereof.
Procedures and/or references for oxidative coupling via boronic acid or other organometallic coupling agents are known to one of skill in the art and may be found in general reference texts, such as for example, Advanced Organic Chemistry by J. March, 4th edition, Wiley-Interscience, New York, NY. Oxidative rearrangement of compound 4 using PDC or PCC in methylene chloride gives 5 which can be cyclized according to the procedure disclosed in Majetich et al . J. Org. Chew. 1997, 62, 6928. Treatment of the cyclized product 6 with an organometallic reagent such as a methylcuperate introduces an alkyl group at the ring junction to give compound 7. Other substitutions can be effected at the 5- position (bridgehead) by use of differently substituted organocuperates or organometallic agents. Nitration can be accomplished using reagents such as cuprous nitrate (Davis et al . Aust . J. Chem . 1987, 40, 1283) , cold fuming nitric acid or a mixture of H2S04 and KN03. Treatment of 8 with BBr3 or pyridine hydrochloride deprotects the methoxy group to give 9.
Compounds of formula I wherein R_.0 is ORa may be obtained by alkylation of the phenol (9) with an alkyl halide or triflate and a base i.e. sodium hydride, sodium ethoxide or lithium isopropoxide .
Alternatively, reaction of 9 with trifluoromethanesulfonic anhydride and pyridine gives 10. Reaction of 10 with carbon monoxide, a palladium catalyst containing a suitable ligand and methanol (Prince, et al . Synlett, 1991, 405) gives 11. Hydrogenation of the nitro group in 11 with Pd/C and hydrogen gives the primary amino compound 12 which can be bisalkylated with THF in the presence of Ti02 as a catalyst to give 13 (Hargis, et al .
Tetrahedron Lett, 1990, 31 , 2991) . Alternatively the amino group in 12 may be reductively aminated with an aldehyde or ketone or the compound 12 is further reacted with an aldehyde or ketone and then reduced to form either a primary or secondary amine respectively. Procedures for reductive amination of aldehydes or ketones are known to one of skill in the art or may be found in general reference texts. Treatment the ketone 13 with hydride reagents such as for example, sodium borohydride gives the final product 14.
Compounds of formula I wherein R10 is CORb may be prepared from the carbethoxy or carbomethoxy analog of compounds 11, 12, 13, or 14 as appropriate, by a ester to ketone conversion reaction such as for example the use of a Grignard (RbMgX) or alkyl or aryl lithium (RbLi) reagent. The reaction is performed preferably in the presence of an aprotic or ethereal reagents such as for example tetrahydrofuran or diethyl ether. Procedures for such conversions are known to one of skill in the art.
Compounds of formula I wherein, R10 is an alkyl group, alkylaryl (i.e. benzyl) or aryl may be prepared by reduction of the corresponding ketone (R_.0 is C0Rb) discussed above .
Compounds of formula I wherein R_.0 is ORa may be obtained by alkylation of the phenol (9) with an alkyl halide or triflate and a base i.e. sodium hydride, sodium ethoxide or lithium isopropoxide .
Compounds of formula I wherein R10 is -CH0HRa may be prepared from compounds wherein R10 is CORa discussed above by a reduction of the ketone function to the corresponding alcohol. The reduction is effected by
procedures known to one of skill in the art such as use of (sodium borohydride) in ethanol at or about room temperature .
The alcohol or (i.e. R10 is CHOHRa) may be dehydrated to form the corresponding alkene or isomers thereof (which may be separated) to afford a compound of formula I wherein R10 is -C2-C8 alkenyl. Dehydration may be accomplished for example by acid promoted elimination of water usually under heating to possibly to reflux, or by dehydrating agents such as phosphorus pentoxide, etc. Methods of effecting such elimination or dehydration are known to one of skill in the art and are disclosed in for example R.C. Larock, Transformations in Organic Synthesis, 4th ed., Wiley-VCH publishers, New York, NY, and references therein.
Alternatively, an alkene functionality may be introduced at R10 by olefination of the ester (11) or its carbethoxy analog to afford an α,β-unsaturated ester at the Rio position. The ester may be olefinated for example by use of the Peterson olefination procedure (Org. React . , 38, 1, (1990) .
Olefination of the aldehyde or ketone functionality (Ri0 = CHO or CORa) may be accomplished for example by applying the Wittig reaction (see Advanced Organic Chemistry by J. March, 3th edition, page 845, Wiley- Interscience, New York, NY, and references therein.
Alternatively, an aldehyde obtained for example from the esters 11, 12, 13 or 14 by reduction methods known to one of skill in the art may be olefinated by use of the Wardsworth-Emmons modification of the Wittig reaction (see
"Wittig Olefination and Modifications Involving Phosphoryl Stabilized Carbanions," Chem. Rev., 89, 863 (1989).
Compounds of formula I wherein R10 is CONR Rι5, may be prepared from compounds 11, 12, 13 or 14 by hydrolysis of the esters to the acid followed by reaction of the incipient acid with ammonia or saturated solution of ammonia or ammonia gas under pressure. Alternatively, the amide, particularly substituted amides may be prepared using the amine HNRι4Rι5 following acid to amide conversion procedures known to one of skill in the art. See for example Weinreb et al . , An alternative procedure for the aluminum-mediated conversion of esters to amides, Syn . Cowm. , 1982, 12, 989- 993, or R.C. Larock, Transformations in Organic Synthesis, 4th ed., Wiley-VCH publishers, New York, NY and references therein.
Compound of Formula I wherein R2 or R2' is a hydroxyl may be prepared using a procedure described in Scheme 2.
Scheme 2
According to Scheme 2, Aldol condensation of commercially available 7-methoxy-l-tetralone and methyl vinyl ketone or substituted methylvinyl ketone in the presence of a base such as NaOEt gives compound 1 of Scheme 2 or substituted analog respectively. Unsaturated ketone 1 or substituted analog thereof, can be reduced to the alcohol 2 using catalytic hydrogenation. Protection of the alcohol 2 with a protecting agent e.g. a silyl protecting group such as t-butyldiphenylsilyl chloride followed by the same transformation procedure described in Scheme 1 yields compound 4 of scheme 2. Deprotection of the silyl group with tetrabutylammonium fluoride (Bu4NF) or aqueous acids sucha s for example aqueous hydrochloric acid, provides alcohol 5.
Compound of Formula I wherein R3 or R3' is a hydroxyl may be prepared using a procedure described in Scheme 3.
Scheme 3
3
According to Scheme 3, Aldol condensation of commercially available 7-methoxy-2-tetralone and methyl vinyl ketone in the presence of a base such as NaOEt gives unsaturated ketone 1 of Scheme 3. 1,4 addition of 1 with LiCu(CH3)2 produces ketone 2. Using the same transformation procedure described in Scheme 1 affords ketone 3. The ketone 3 can be further reduced with NaBH4 or other hydrides or reducing agents to afford the desired alcohol 4 of Scheme 3.
Compound of Formula I wherein R4 or R4' is a hydroxyl may be prepared using a procedure described in Scheme 4.
Scheme 4
Alkylation of 7-methoxy-2-tetralone with 1-bromo- 5-methyl-5-hexene gives compound 1 of Scheme 4. Ozonolysis of 1 (Scheme 4) followed by intramolecular aldol condensation yields ketone 4. Procedures for ozonolysis are known to one of skill in the art and may be found in general reference texts described earlier. Procedures to effect aldol condensation are known to one of skill in the art. Reduction of the unsaturated ketone 4 and dehydration of the resulting alcohol produces olefin 5. Further ozonolysis of 5 affords ketone 6. The same transformation procedure described in scheme 1 (beginning with compound 6) or scheme 3 (beginning with compound 2) may be applied to compound 6 of Schemem 4 to afford alcohol 7 of Scheme 4.
Examples
The following examples are provided for the purpose of illustrating the compounds of the current invention and are not intended to limit its scope.
Example 1
Trans-8 -Hydroxy- 9-hydro- 1, 2- [a,b] [ (l-carboxyethyl-2-N- pyrolidinyl) benzo-4 , 5-yl] -cis-10-methyldecalin also named 5- Hydroxy-8a-methyl-2-pyrrolidin-l-yl-4b, 5,6,7,8, 8a, 9, 10- octahydro-phenanthrene-3 -carboxylic acid ethyl ester, and available from Aldrich as Compound #95709 Aldrich Chemical Co . , USA.
Compound of example 1 is a white crystalline powder with the following physical chemical characteristics:
PMR: 7.40δ (lH,s); 6.55δ (lH,s); 4.40δ (2H,q); 4.05δ (lH,s);
3.28δ (3H,m) ; 2.55δ (lH,dd) ; 2.33δ (lH,s) ; 2.05-1.80δ (6H,m) ;
1.58δ (4H,m) ; 1.40δ (6H,t,m) ; 1.15δ (lH,d) ; l.OOδ (lH,s) ;
0.82δ (3H,s) (CDC13) .
MS: e/z=358.3 (M+l) ES+ consistent with C22H3ιN03
Further NMR experiments using XH, 13C, COSY, or ROESY, and
EHSQC techniques, determined that the angular methyl and hydrogen groups at the ring juncture of the decalin are located on the same side of each other, i.e., cis. Also,
the hydroxyl function on the 8 -position of the decalin ring system is on the opposite side from the angular hydrogen at the 10-position.
The racemic mixture (41 mg) from Example 1 above, was further purified using a Chiralpak AD column (0.46 X25 cm) and eluted with a mixture of isopropyl alcohol (20%) and heptane (80%) at flow rate of 1.0 mL/min to give two enantiomers, A and B. Enantiomer 1A has shown EC50 4 μM, while enantiomer IB is less active in the ABC1 bDNA assay. The characteristics of these two enantiomers are as follows:
Enantiomer 1A: 14.8 mg, 90% ee (enantimeric excess), t = 6.664 min, [α] 24 D -25.81° (c 1.24, CH2C12) .
Enantiomer IB: 15.1 mg, 90% ee, t = 7.263 min, [ ] 24 D +25.19° (c 1.35, CH2C12) .
Example 2
8-Keto-l,2-[a,b] [ (1-carboxyethyl-l-N-pyrolidinyl) benzo-4 , 5- yl] -10-methyldecalin also named 8a-Methyl-5-oxo-2- pyrrolidin-l-yl-4b, 5, 6, 7, 8, 8a, 9, 10-octahydro-phenanthrene-3- carboxylic acid ethyl ester
Compound of example 2 is a white crystalline powder with the following physical chemical characteristics:
PMR: 7.18δ (lH,s); 6.60δ (lH,s); 4.3lδ (2H,q); 3.28δ (4H,m); 2.95δ (lH,m); 2.85δ (lH,m); 2.35δ (2H,m); 2.10-1.65δ (8H,m); 1.57δ (2H,s); 1.45δ (lH,dd); 1.35δ (3H,t); 1.05δ (3H,s) CDC13. MS: e/z=356.1 (M+l) ES+, consistent with C22H29N03.
Example 3
8-Hydroxy-l, 2- [a,b] [ (1-hydroxymethyl-l-N-pyrolidinyl) benzo- 4, 5-yl] -10-methyldecalin also named 6 , 10a-Dimethyl-7- pyrrolidin-l-yl-1, 2, 3, 4, 4a, 9, 10, 10a-octahydro-phenanthren-4- ol , also named 6-Hydroxymethyl-10a-methyl-7-pyrrolidin-l-yl- l,2,3,4,4a,9,10, 10a-octahydro-phenanthren-4-ol
Compound of example 3 is a white crystalline powder with the following physical chemical characteristics:
PMR: 7.10δ (lH,s); 7.05δ (lH,s); 4.95δ (2H, s) ; 4.38δ (2H,m);
4.08δ. (2H,m); 3.45δ (2H,m); 2.8lδ (2H,dd); 2.65-2.30δ
(5H,m) ; 1.95δ (lH,d) ; 1.86δ (lH,t) ; 1.67δ (3H,m) ; 1.48δ
(3H,m) ; 1.25δ (lH,d) ; 0.81δ (3H,s) CDCl3. MS: e/z=316.3 (M+l) ES+ consistent with C2oH29N02.
Example 4
8-Hydroxy-l , 2- [a, b] [ (1-methyl -1-N-pyrolidinyl) benzo-4 , 5-yl] - lO-methyldecalin, also named 6, 10a-Dimethyl-7-pyrrolidin-l- yl-1,2, 3, 4, 4a, 9, 10, 10a-octahydro-phenanthren-4-ol
Compound of example 4 is a white crystalline powder with the following physical chemical characteristics:
PMR: 6.85δ (lH,s); 6.62δ (lH,s); 4.03δ (2H, s) ; 3.23δ (3H, m)- ; 2.76δ (2H,m); 2.50δ (lH,m); 2.28δ (3H,m); 2.00-1.70δ (5H,m); 1.55δ (3H,s); 1.4lδ (2H,t); l.lOδ (lH,d); l.OOδ (lH,m); 0.83δ (3H,s) CDC13. MS: e/z=300.3 (M+l) ES+
E/z=299.2 (M) FD consistent with C2oH29NO.
ASSAY
The following assay protocol and result (s) thereof demonstrating the utility and efficacy of the methods of the current invention are given for the purpose of illustration and are not meant to be limiting in any way.
bDNA Assay for ABC1 mRNA
Cell incubation and treatment Human monocytic THP-1 cells were plated in 96 well plate at the density of 1 x 105/well on day 0 and treated
with phorbol 12-myristate 13-acetate at 10 nM for 48 hours. The medium was then changed to DMEM with 10% lipoprotein deficient serum and treated with compounds at various concentrations for 24 hours. Cells were then lysed and mRNA measured as described.
The bDNA assay was performed according to the manufacturer protocol for the QuantiGene High Volume Kit (Bayer Diagnostics) . After the challenge of the cells with a compound of the invention, cells were lysed with QuantiGene lysis buffer containing the ABC1 mRNA oligonucleotides described below. The ABCl oligonucleotides (capture extenders (CEs) , label extenders (LEs) , and blockers (BLs) ) are shown in Table 1.
After a 15 minute incubation at 37°C, lOOμl of the lysis buffer from each well were transferred to the corresponding wells of the capture plate. The capture plate was incubated overnight at 53°C. The capture plate was then washed twice with QuantiGene () wash buffer followed by addition of lOOμl/well QuantiGene amplifier working reagent. The plate was incubated for 60 minutes at 46°C followed by two washes. The mRNA to be measured was then labeled by addition of lOOμl QuantiGene label probe working buffer followed by a 60 minute incubation at 46°C. The capture plate was then washed twice followed by addition of lOOμl/well QuantiGene substrate plus QuantiGene enhancer reagent. The plates were incubated at 37°C for up to 30 minutes and then read on a luminometer to detect the luminescent signal. The induction of ABCl mRNA expression was calculated as a ratio of treated luminescent levels compared to untreated control levels. An EC50 values i.e. effective response relative to maximal; response was calculated. Percent efficiency i.e. the percentage efficacy
relative to a lOuM 22r hydroxyxholesterol (a well characterized LXR standard known to one of skill in the art) was also calculated.
Results :
Compound Example 1 EC50 3.895uM
%Eff 75.40
Table 1 bDNA oligonucleotides for detection and quantitation of human ABCl mRNA
Method of Treatment
As used herein, the term "effective amount" means an amount of compound of the present invention, i.e., formula I, which is capable of alleviating the symptoms of the various pathological conditions herein described. The specific dose of a compound administered according to this invention will, of course, be determined by the particular circumstances surrounding the case including, for example, the compound administered, the route of administration, the state of being of the patient, and the pathological condition being treated. A typical daily dose will contain a nontoxic dosage level of from about 1 mg to about 1000 mg/day of a compound of the present invention. Preferred daily doses generally will be from about 5 mg to about 250 mg/day.
The compounds of this invention can be administered by a variety of routes including oral, rectal, transdermal, subcutaneous, intravenous, intramuscular, and intranasal . These compounds preferably are formulated prior to administration, the selection of which will be decided by the attending physician. Thus, another aspect of the present invention is a pharmaceutical composition comprising an effective amount of a compound of Formula I, or a pharmaceutically acceptable salt thereof, solvate, prodrug, enantiomer or prodrug thereof, and a pharmaceutically acceptable carrier, diluent, or excipient.
The total active ingredients in such formulations comprises from 0.1% to 99.9% by weight of the formulation. By "pharmaceutically acceptable" it is meant the carrier, diluent, excipients and salt must be compatible with the other ingredients of the formulation, and not deleterious to the recipient thereof .
Pharmaceutical formulations of the present invention can be prepared by procedures known in the art using well-known and readily available ingredients. For example, the compounds of formula I can be formulated with common excipients, diluents, or carriers, and formed into tablets, capsules, suspensions, powders, and the like. Examples of excipients, diluents, and carriers that are suitable for such formulations include the following: fillers and extenders such as starch, sugars, mannitol, and silicic derivatives; binding • agents such as carboxymethyl cellulose and other cellulose derivatives, alginates, gelatin, and polyvinyl -pyrrolidone; moisturizing agents such as glycerol ; disintegrating agents such as calcium carbonate and sodium bicarbonate; agents for retarding dissolution such as paraffin; resorption accelerators such as quaternary ammonium compounds; surface active agents such as cetyl alcohol, glycerol monostearate; adsorptive carriers such as kaolin and bentonite; and lubricants such as talc, calcium and magnesium stearate, and solid polyethyl glycols.
The compounds also can be formulated as elixirs or solutions for convenient oral administration or as solutions appropriate for parenteral administration, for example, by intramuscular, subcutaneous or intravenous routes. Additionally, the compounds are well suited to formulation as sustained release dosage forms and the like. The formulations can be so constituted that they release the active ingredient only or preferably in a particular physiological location, possibly over a period of time. The
coatings, envelopes, and protective matrices may be made, for example, from polymeric substances or waxes.
Compounds of formula I, generally, will be administered in a convenient formulation as determined by the attending physician. The following formulation examples are only illustrative and are not intended to limit the scope of the present invention.
Formulations In the formulations which follow, "active ingredient" means a compound of formula I, a salt solvate, racenate, enantiomer or prodrug thereof.
Formulation 1: Gelatin Capsules
Hard gelatin capsules are prepared using the following:
Ingredient Quantity (mg/capsule) Active ingredient 0.1 - 1000
Starch, NF 0 - 650
Starch flowable powder 0 - 650
Silicone fluid 350 centistokes 0 - 15
The formulation above may be changed in compliance with the reasonable variations provided.
A tablet formulation is prepared using the ingredients below:
Formulation 2 : Tablets
Ingredient „ Quantity (mg/tablet)
Active ingredient 2.5 - 1000
Cellulose, microcrystalline 200 - 650
Silicon dioxide, fumed 10 - 650 Stearate acid 5 - 15
The components are blended and compressed to form tablets.
Alternatively, tablets each containing 2.5 - 1000 mg of active ingredient are made up as follows:
Formulation 3 : Tablets
Ingredient Quantity (mg/tablet]
Active ingredient 25 - 1000
Starch 45
Cellulose, microcrystalline 35
Polyvinylpyrrolidone 4
(as 10% solution in water)
Sodium carboxymethyl cellulose 4.5
Magnesium stearate 0.5
Talc 1
The active ingredient, starch, and cellulose are passed through a No. 45 mesh U.S. sieve and mixed thoroughly. The solution of polyvinylpyrrolidone is mixed with the resultant powders which are then passed through a No. 14 mesh U.S. sieve. The granules so produced are dried at 50°-60° C and passed through a No. 18 mesh U.S. sieve. The sodium carboxymethyl starch, magnesium stearate, and talc, previously passed through a No. 60 U.S. sieve, are then added to the granules which, after mixing, are compressed on a tablet machine to yield tablets.
Suspensions each containing 0.1 - 1000 mg of medicament per 5 ml dose are made as follows:
Formulation 4 : Suspensions
Ingredient Quantity (mg/5 ml)
Active ingredient 0.1 - 1000 mg
Sodium carboxymethyl cellulose 50 mg
Syrup 1.25 mg
Benzoic acid solution 0.10 mL
Flavor q.v.
Color q.v.
Purified water to 5 L
The medicament is passed through a No. 45 mesh U.S. sieve and mixed with the sodium carboxymethyl cellulose and syrup to form a smooth paste. The benzoic acid solution, flavor, and color are diluted with some of the water and added, with stirring. Sufficient water is then added to produce the required volume.
An aerosol solution is prepared containing the following ingredients :
Formulation 5: Aerosol
Ingredient Quantity (% by weight) Active ingredient 0.25
Ethanol 25.75
Propellant 22 (Chlorodifluoromethane) 70.00
The active ingredient is mixed with ethanol and the mixture added to a portion of the propellant 22, cooled to 30° C, and transferred to a filling device. The required amount is then fed to a stainless steel container and diluted with the remaining propellant. The valve units are then fitted to the container.
Formulation 6: Intravenous Solution
Ingredient Quantity
Active ingredient 50 mg
Isotonic saline 1,000 mL
The solution of the above ingredients is intravenously administered to a patient at a rate of about 1 mL per minute .