WO2001077077A1

WO2001077077A1 - Substituted (hetero)aryl carboxamide derivatives as microsomal triglyceride transfer protein (mtp) and apolipoprotein b (apo b) secretion

Info

Publication number: WO2001077077A1
Application number: PCT/EP2001/004052
Authority: WO
Inventors: Robert E. Ii Damon
Original assignee: Novartis Ag; Novartis-Erfindungen Verwaltungsgesellschaft M.B.H.
Priority date: 2000-04-10
Filing date: 2001-04-09
Publication date: 2001-10-18
Also published as: AU2001262185A1

Abstract

Compounds of formula (I) wherein R1, L, L', alk, W and Z have the meanings as defined hereinbefore; and pharmaceutically acceptable salts thereof; are useful as inhibitors of microcosmal triglyceride transfer protein (MTP) and of apolipoprotein B (apo B) secretion.

Description

Organic Compounds

The present invention relates to the amide derivatives described herein which are particularly useful as inhibitors of microsomal triglyceride transfer protein (MTP) and of apolipoprotein B (apo B) secretion, methods for preparation thereof, pharmaceutical compositions comprising said compounds, a method of inhibiting MTP and apo B secretion and of treating conditions in mammals which are responsive to MTP inhibition or inhibition of apo B secretion using said compounds or pharmaceutical compositions comprising said compounds of the invention.

The invention relates to the compounds of formula I

O R,— L— C— NH — L'— V — alk— W — Z (I)

wherein

Ri is aryl, cycloalkyl, heterocyclyl, aryl-lower alkoxy or aryl-lower alkylthio; provided that Ri is not carboxyphenyl when both V and W are O or when one of V and W is O and the other is a direct bond, and when Z is aryl or heteroaryl;

L and L' are arylene or heteroarylene;

V and W are independently O, S(O)_m, NR₀ or a direct bond, provided that only one of V and may be a direct bond;

Z is aryl or heteroaryl; or when W is NR₀, Z is also

_/ . heteroaryl-lower alkyl,—- COR_a,-CON , — COOR_d, or — SO₂R_θ in which

Re

R_a, R_d and R_e are independently optionally substituted lower alkyl, cycloalkyl, adamantyl, aryl, heteroaryl or (aryl or heteroaryl)-lower alkyl; and Rb and R_c are independently hydrogen, cycloalkyl, optionally substituted lower alkyl, aryl, heteroaryl or (aryl or heteroaryl)-lower alkyl; or R and R₀ together represent lower alkylene or lower-alkylene interrupted by O, S or N-(H, lower alkyl, acyl or aralkyl); R₀ is hydrogen, lower alkyl or aryl-lower alkyl; m is zero, 1 or 2; and alk is lower alkylene; and pharmaceutically acceptable salts thereof.

More particularly, the invention relates to compounds of formula I wherein R^ is monocyclic_. (aryl, heteroaryl, or aryl-lower alkoxy); and L and L' are monocyclic (arylene or heteroarylene).

A particular embodiment of the invention relates to the compounds of formula II

wherein

R is hydrogen, lower alkyl, lower alkoxy, halo or trifluoromethyl;

Ri is aryl, cycloalkyl, heterocyclyl, aryl-lower alkoxy or aryl-lower alkylthio; provided that R_t is not carboxyphenyl when V and W are O or when one of V and W is O and the other is a direct bond, and when Z is aryl or heteroaryl;

L is arylene or heteroarylene;

Y is CH or N;

V and W are independently O, S(O)_m, NR₀ or a direct bond, provided that only one of V and W may be a direct bond;

Z is aryl or heteroaryl; or when W is NR₀, Z is also

heteroaryl-lower — COOR_d, or — SO₂R in which

R_a» R and R_β are independently optionally substituted lower alkyl, cycloalkyl, adamantyl, heteroaryl or (aryl or heteroaryl)-lower alkyl; and R_b and R₀ are independently hydrogen, cycloalkyl, optionally substituted lower alkyl, aryl, heteroaryl or (aryl or heteroaryl)-lower alkyl; or R_b and R₀ together represent lower alkylene or lower-alkylene interrupted by O, S or N-(H, lower alkyl, acyl or aralkyl);

R₀ is hydrogen, lower alkyl or aryl-lower alkyl; m is zero, 1 or 2; and n is an integer from 1-4 provided that either V or W is a direct bond when n is 1 ; and pharmaceutically acceptable salts thereof.

Another particular embodiment of the invention relates to the compounds of formula III

wherein X is N or CR₂;

Y is N or CH;

R is hydrogen, lower alkyl, lower alkoxy, halo or trifluoromethyl;

R₀ is hydrogen or lower alkyl;

Ri is aryl, cycloalkyl, heteroaryl, aryl-lower alkoxy or aryl-lower alkylthio; provided that Ri is not carboxyphenyl when both V and W are O or when one of V and W is O and the other is a direct bond;

Ar is monocyclic aryl or heteroaryl;

R₂, R₃, R4, and R₅ are independently hydrogen, optionally substituted alkyl, halo, amino, substituted amino, trifluoromethyl, cyano, carboxyl, alkoxycarbonyl, aralkoxycarbonyl, (alkyl, aryl or aralkyl)-thio, (alkyl, aryl, or aralkyl)-oxy, acyloxy, (alkyl, aryl or aralkyl)-aminocarbonyloxy; or any two or R₂, R3, R and R₅ at adjacent positions are alkylenedioxy; n is 2 or 3; m is zero; and pharmaceutically acceptable salts thereof.

Preferred are said compounds of formula III wherein V is O, S(O)_m or NR₀, and W is a direct bond; and pharmaceutically acceptable salts thereof.

Particularly preferred are the compounds of formula III wherein Ri is phenyl or phenyl-lower alkoxy, each optionally substituted on phenyl by lower alkyl, lower alkoxy, halo, trifluoromethyl or cyano; X is CR₂; R, R₂, R3, R4 and R₅ are independently hydrogen, lower alkyl, lower alkoxy, halo or trifluoromethyl; Y is CH or N; V is NH, N-CH₃ or O; n is 1 , 2 or 3; W is a direct bond; and Ar is phenyl or phenyl substituted by lower alkyl, halo, trifluoromethyl, lower alkoxy or cyano; or Ar is thienyl, pyridyl, indolyl or pyrimidyl optionally substituted by lower alkyl; and pharmaceutically acceptable salts thereof.

Further preferred are the compounds of formula III wherein Ri is trifluoromethylphenyl or benzyloxy; Y is CH or N; X is CR₂; R₂ is methyl; R is hydrogen, methyl, or chloro; R₃, R₄ and R₅ are hydrogen; V is NH; n is 2 or 3; W is a direct bond; Ar is phenyl or 2-pyridyl; and pharmaceutically acceptable salts thereof.

Another particular embodiment of the invention relates to the compounds of formula IV

wherein X is N or CR₂;

Y is N or CH;

V is O, S(O)_m, NR₀ or a direct bond;

R is hydrogen, lower alkyl, lower alkoxy, halo or trifluoromethyl;

R₀ is hydrogen or lower alkyl;

R_! is aryl, cycloalkyl, heteroaryl, aryl-lower alkoxy or aryl-lower alkylthio; R₂, R₃, R₄, and R₅ are independently hydrogen, lower alkyl, halo, trifluoromethyl, cyano, (lower alkyl, aryl, or aralkyl)-thio, (lower alkyl, aryl, or aralkyl)-oxy, acyloxy, (alkyl, aryl or aralkyl)-aminocarbonyloxy; or any two or R₂, R₃, R₄ and R₅ at adjacent positions are alkylenedioxy;

Z is heteroaryl-lower alkyl, — COR_a,-CON^' , — COOR_d, or — SO₂R_e in which

Re

R_a, R_d and R_e are independently optionally substituted lower alkyl, cycloalkyl, adamantyl, aryl, heteroaryl or (aryl or heteroaryl)-lower alkyl; and R and R_c are independently hydrogen, cycloalkyl, optionally substituted lower alkyl, aryl, heteroaryl or (aryl or heteroaryl)-lower alkyl; or R and R_c together represent lower alkylene or lower-alkylene interrupted by O, S or N-(H, lower alkyl, acyl or aralkyl); m is zero, 1 or 2; and n is an integer from 1-4; and pharmaceutically acceptable salts thereof.

Preferred are the compounds of formula IV wherein Ri is phenyl or phenyl -lower alkoxy, each optionally substituted on phenyl by lower alkyl, lower alkoxy, halo, trifluoromethyl or cyano; X is CR₂; Y is CH or N; R, R₂, R₃, R₄ and R₅ are independently hydrogen, lower alkyl, lower alkoxy, halo or trifluoromethyl; R₀ is hydrogen; V is a direct bond, NH, N-CH₃ or O; n is 2 or 3; Z is heteroaryl-

lower alkyl, — , — COOR_d or — SO₂R_θ;

R_b and R_c are lower alkyl; or R and R_c together represent lower alkylene or lower alkylene interrupted by O, S, N-(H, lower alkyl or aralkyl);

R_d is lower alkyl or aryl -lower alkyl;

-Rβ-is-lαwer-alkyl_^TrjonoxxyOlic^ryl-oii-morjo and pharmaceutically acceptable salts thereof.

Further preferred are the compounds of formula IV wherein R is hydrogen, methyl or chloro; Ri is trifluoromethylphenyl; X is CR₂; Y is CH or N; R₂ is methyl; R₃, FU and R₅ are hydrogen; Ro is hydrogen; V is NH, O, or a direct bond; n is 2 or 3; and Z is -COOR_d wherein R_d is lower alkyl; and pharmaceutically acceptable salts thereof.

Listed below are definitions of various terms used to describe the compounds of the instant invention. These definitions apply to the terms as they are used throughout the specification (unless they are otherwise limited in specific instances either individually or as part of a larger group).

The term "lower" referred to herein in connection with organic radicals or compounds respectively generally defines, if not defined differently, such with up to and including 7, preferably up and including 4 and advantageously one or two carbon atoms. Such may be straight chain or branched.

The term "optionally substituted lower alkyl" refers to unsubstituted or substituted straight or branched chain hydrocarbon groups having 1 to 7 carbon atoms. Exemplary unsubstituted alkyl groups include methyl, ethyl, propyl, isopropyl, n-butyl, t-butyl, isobutyl, pentyl, hexyl, isohexyl, heptyl, 4,4-dimethylpentyl, and the like.

The term "substituted lower alkyl" refers to alkyl groups substituted by one or more of the following groups: halo (such as CCI₃ or CF₃), hydroxy, alkoxy, alkoxyalkoxy, aryloxy, cycloalkyl, alkanoyl, alkanoyloxy, amino, substituted amino, alkanoylamino, thiol, alkylthio, arylthio, alkylthiono, alkylsulfonyl, arylsulfonyl, heteroarylsulfonyl, aminosulfonyl, nitro, cyano, carboxy, carbamyl, alkoxycarbonyl, aryl, aralkoxy, heterocyclyl (e.g., indolyl, imidazolyl, furyl, thienyl, thiazolyl, pyrrolidyl, pyridyl, pyrimidyl), fluorenyl, and the like.

The term "lower alkyl" refers to those alkyl groups as described above having 1 to 7, preferably 1 to 4 carbon atoms.

The term "halogen" or "halo" refers to fluorine, chlorine, bromine and iodine, preferably

The term "lower alkylene" refers to a straight chain bridge of 1 to 7 carbon atoms connected by single bonds (e.g., -(CH₂)χ- wherein x is 1 to 7) which may be substituted with 1 to 3 lower alkyl groups. The term "lower alkylene interrupted by O, S, N-(H, alkyl, acyl or aralkyl)" refers to a straight chain of 2 to 6 carbon atoms which is interrupted by O, S, N-(H, alkyl, acyl or aralkyl), such as (m)ethyleneoxy(m)ethylene, (m)ethylenethio(m)ethylene, or (m)ethyleneimino(m)ethylene.

The term "cycloalkyl" refers to cyclic hydrocarbon groups of 3 to 8 carbon atoms such as cyclopentyl, cyclohexyl or cycloheptyl.

The term "alkoxy" or "alkyloxy" refers to alkyl-O-.

The term "alkanoyl" refers to alkyl-C(O)-.

The terms "alkylamino" and "dialkylamino" refer to (alkyl)NH- and (alkyl)2N-, respectively.

The term "alkanoylamino" refers to alkyl-C(O)-NH-.

The term "alkylthio" refers to alkyl-S-.

The term "alkoxycarbonyl" refers to alkyl-O-C(O)-.

The term "acyl" refers to alkanoyl, aroyl, heteroaroyl, aryl-alkanoyl, heteroarylalkanoyl, and the like.

The term "aryl" or "ar", refers to monocyclic or bicyclic aromatic hydrocarbon groups having 6 to 12 carbon atoms in the ring portion, such as phenyl, naphthyl, tetrahydronaphthyl, and biphenyl groups, each of which may optionally be substituted by one to four substituents such as alkyl, halo, trifluoromethyl, hydroxy, alkoxy, halo-alkyl, alkanoyl, alkanoyloxy, amino, substituted amino, alkanoylamino, thiol, alkylthio, nitro, cyano, carboxy, carboxyalkyl, carbamyl, alkoxycarbonyl, alkylthiono, alkylsulfonyl, aminosulfonyl, heterocyclyl and the like.

The term "aralkoxy" refers to an aryl group linked to an alkoxy group, such as benzyloxy.

The term "arylsulfonyl" refers to aryl-SO₂-. The term "aroyl" refers to aryl-CO-.

The term "heterocyclyl" refers to an optionally substituted, fully saturated or unsaturated, aromatic or nonaromatic cyclic group, for example, which is a 4 to 7 membered monocyclic, 7 to 11 membered bicyclic system, which has at least one heteroatom in at least one carbon atom-containing ring. Each ring of the heterocyclic group containing a heteroatom may have 1 , 2 or 3 heteroatoms selected from nitrogen atoms, oxygen atoms and sulfur atoms, where the nitrogen and sulfur heteroatoms may also optionally be oxidized. The heterocyclic group may be attached at any heteroatom or carbon atom.

Exemplary monocyclic heterocyclic groups include pyrrolidinyl, pyrrolyl, pyrazolyl, oxetanyl, pyrazolinyl, imidazolyl, imidazolinyl, imidazolidinyl, oxazolyl, oxazolidinyl, isoxazolinyl, isoxazolyl, thiazolyl, thiadiazolyl, thiazolidinyl, isothiazolyl, isothiazolidinyl, fury!, tetrahydrofuryl, thienyl, oxadiazolyl, piperidinyl, piperazinyl, 2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxopyrrolodinyl, 2-oxoazepinyl, azepinyl, 4-piperidonyl, pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, tetrahydropyranyl, morpholinyl, thiamorpholinyl, thiamorpholinyl sulf oxide, thiamorpholinyl sulfone, 1 ,3-dioxolane and tetrahydro-1,1-dioxothienyl, and the like.

Exemplary bicyclic heterocyclic groups include indolyl, benzothiazolyl, benzoxazolyl, benzothienyl, quinuclidinyl, quinolinyl, tetrahydroisoquinolinyl, isoquinolinyl, benzimidazolyl, benzopyranyl, indolizinyl, benzofuryl, chromonyl, coumarinyl, benzopyranyl, cinnolinyl, quinoxalinyl, indazolyl, pyrrolopyridyl, furopyridinyl (such as furo[2,3-c]pyridinyl, furo[3,2-b]pyridinyl] or furo[2,3-b]pyridinyl), dihydroisoindolyl, dihydroquinazolinyl (such as 3,4-dihydro-4-oxo-quinazolinyl) and the like.

The term "heterocyclyl" also includes substituted heterocyclic groups. Substituted heterocyclic groups refer to heterocyclic groups substituted independently with 1 , 2 or 3 of e.g. the following:

(a) lower alkyl;

(b) — hydroxy-(or-protected hydroxy)

(c) halo;

(d) oxo (i.e. = O);

(e) amino or substituted amino;

(f) lower alkoxy; or

(g) cyano; The term "heteroaryl" "or heteroar" refers to an aromatic heterocycle, for example monocyclic or bicyclic aryl, such as pyrrolyl, pyrazolyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, furyl, thienyl, pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, indolyl, benzothiazolyl, benzoxazolyl, benzothienyl, quinolinyl, isoquinolinyl, benzimidazolyl, benzofuryl, and the like, optionally substituted by e.g., lower alkyl, lower alkoxy or halo. The point of attachment of the heteroaryl group is at any of the available carbon positions.

The term "pyridyl" refers to 2-,3- or 4-pyridyl.

The term "heteroaroyl" refers to heteroaryl-CO-. .

The term "acylamino" refer to acyl-NH-.

The term "substituted amino" refers to amino mono- or, independently, disubstituted by alkyl, aralkyl, aryl, heteroaryl, cycloalkyl, cycloalkylalkyl, heteroaralkyl, or disubstituted by lower alkylene or lower alkylene interrupted by O, S, N-(H, alkyl, acyl or aralkyl), and the like.

The term "heteroarylene" for "L" refers to a monocyclic or bicyclic heteroaryl linking group preferably pyridylene, thienylene, furanylene, each optionally substituted by lower alkyl, lower alkoxy, halo or cyano with the points of attachment of the two groups in formula I attached thereto being preferably at adjacent carbon atoms, such as 2,3-pyridylene, 2,3- thienylene or 2,3-furanylene.

The term "arylene" for the linking group L refers to a monocyclic or bicyclic aryl linking group, preferably phenylene, advantageously ortho-phenylene optionally substituted by 1 to 4 substituents independently selected from lower alkyl, lower alkoxy, halo, trifluoromethyl and cyano, or arylene preferably refers to phenylene optionally substituted at adjacent positions by alkylenedioxy.

The term "heteroarylene" for "L* " refers to a monocyclic or bicyclic heteroaryl linking group preferably pyridylene, thienylene, furanylene, each optionally substituted by e.g. lower alkyl, lower alkoxy, halo or cyano, with the points of attachment of the two groups in formula I attached thereto being preferably at non-adjacent carbon atoms, such as pyridylene, preferably 2,4- or 2,5-pyridylene.

The term "arylene" for the linking group L' refers preferably to phenylene, advantageously para-phenylene optionally substituted by 1 to 4 substituents independently selected from lower alkyl, lower alkoxy, halo, trifluoromethyl or cyano, or arylene refers to phenylene optionally substituted at adjacent positions by alkylenedioxy.

Pharmaceutically acceptable salts are primarily acid addition salts, such as of mineral acids, organic carboxylic, and organic sulfonic acids e.g., hydrochloric acid, methanesulfonic acid, maleic acid, provided a basic group, such as amino or pyridyl, constitutes part of the structure.

Pharmaceutically acceptable salts of any acidic compounds of the invention are salts formed with bases, namely cationic salts such as alkali and alkaline earth metal salts, such as sodium, lithium, potassium, calcium, magnesium, as well as ammonium salts, such as ammonium, trimethylammonium, diethylammonium, and tris-(hydroxymethyl)- methylammonium salts.

The compounds of the invention depending on the nature of the substituents, may possess one or more asymmetric carbon atoms, and therefore may exist as racemates and the (R) and (S) enantiomers thereof. All are within the scope of the invention.

The compounds of the invention of formula I are prepared by condensing a compound of the formula

RrL-COOH (V)

-or--a -eacliv^-deιiv-atiy-e he^ Ri and L have meaning as defined above, with an amine of the formula

NHa-L'-V-alk-W-Z (VI)

wherein L', V, alk, W and Z have meaning as defined hereinabove. The condensation, using a carboxylic acid, is carried out under conditions well known in the art for the preparation of amides, and as illustrated in the examples, in the presence of condensing agents such as 1-ethyl-3-(dimethylaminopropyl)carbodiimide hydrochloride (EDCI) optionally in combination with 1 -hydroxy-7-azabenzotriazole (HOAt). In case a reactive derivative of the acid, e.g. an acid chloride is the starting material, the condensation is carried out in the presence of a base such as N-methylmorpholine, diisopropylethylamine or pyridine.

The substituted aryl carboxylic acids of formula V are prepared as follows.

The acids wherein Ri represents aryl or heteroaryl and L is arylene or heteroarylene are generally prepared by a palladium catalyzed aryl- aryl coupling reaction, e.g. as described in Bioorg. Med. Chem. Lett. 7 (13), 1595 (1997).

For example, acids of formula VII wherein Ri is aryl or heteroaryl and R₂ - R₅ have meaning as defined above for compounds of formula III, are prepared as exemplified in the scheme below by palladium catalyzed coupling of aryl boronic acids of formula VIII with bromo-, iodo- or trifluromethanesulfonoxy- substituted arylcarboxylic acid esters e.g., of formula IX. Subsequent hydrolysis of the ester group of resulting compounds of formula X gives the carboxylic acids of formula VII.

(IX) (VIII) (X) (VII)

Reactive derivatives of the carboxylic acids, e.g. acid chloride, are prepared by reaction with a chlorinating agent, such as oxalyl chloride. The carboxylic acids of formula V wherein Ri represents aryl-lower alkoxy or aryl-lower alkylthio are prepared from the appropriate phenols or thiophenols by reaction with e.g. an aryl-lower alkyl halide e.g. as illustrated below.

The amine starting materials of formula VI are prepared according to general methods known in the art, e.g. as illustrated below and in the example herein.

Illustrative thereof the amines of formula XI

(XI)

wherein R, V, Y, W, n and Ar have meaning as defined for compounds of formula III are prepared by reducing the corresponding nitro compounds of formula XII

(XII) under conditions well known in the art, e.g. by catalytic hydrogenation.

Nitro intermediates of formula XII, e.g. those wherein V is NR₀ and W is a direct bond are prepared by condensing a reactive aromatic nitro derivative of formula XIII

(XIII) (XIV)

with an amine of formula XIV in the presence of a base, e.g. a tertiary amine such as triethylamine, in a polar solvent such as ethanol or dimethylsulfoxide.

Similarly, nitro intermediates of formula XII wherein V is O or S are prepared by condensing e.g. a compound of formula XIII with the appropriate alcohol or thiol in the presence of a base, such as sodium hydride, in a solvent, such as dimethylformamide.

Nitro intermediates of formula XII wherein V is O and W is a direct bond can also be prepared by condensing a nitro compound of formula XV with the appropriate alcohol in the presence

(XV) ©f riphenylphosphine^ncWiethyl-azodicarbc^late^ is O and V is a direct bond are similarly prepared.

The amine starting materials for the preparation of compounds of the invention represented by formula IV, namely the amines of formula XVI

(XVI)

wherein Y, V, R_OJ n and Z have meaning as defined for compounds of formula IV, are also prepared by reduction of the corresponding nitro compounds of the formula XVII

e.g. by catalytic hydrogenation.

The nitro compounds of formula XVII are in turn prepared by condensation of a reactive aromatic nitro derivative, e.g. of formula XIII, with a compound of formula XVIII

VH - (CH₂)_n - NRo - Z (XVIII)

wherein V is O, S or NR₀,

and Z is e.g.— COR_a, — COORd or — SO₂ R_e as defined above.

The condensation is carried out in the presence of a base. When V is NR₀, the condensation is preferably carried^'oT.tlrrth^TB^Ti ϋ e^gT^ triethylamine, or sodium acetate, using a polar solvent such as ethanol. When V is O or S, the condensation is preferably carried out using a strong base, such as sodium hydride, in a solvent such as dimethylformanide. When V is O, intermediates of formula XVII can also be prepared by condensation of a nitro compound of formula XV with an alcohol of formula XVIII (V being O) in the presence of triphenylphosphine and diethyl azodicarboxylate.

Alternately, the nitro intermediates of formula XVII can be prepared by treating an amine of the formula

with a reactive form of a compound of the formula ZH, e.g. with a compound of the formula Z-CI, wherein Z has meaning as defined above for the intermediates of formula XVIII, in the presence of a base, such as triethylamine, in a polar solvent such as dimethylformamide. Certain of the amine intermediates of formula XIX may be prepared by condensation of starting materials of formula XIII or XV with an appropriate compound of the formula HV- (CH₂)_n NHR₀.

In general, the amines of formula XIX can be prepared by N-deprotection of compounds of formula XVII wherein Z is t-butoxycarbonyl under conditions well known in the art for the removal of a t-butoxycarbonyl group, e.g. with trifluoroacetic acid in methylene chloride.

Alternatively, compounds of the invention represented by formula IV can be prepared from compounds of formula IV wherein Z is t-butoxycarbonyl, prepared as described herein. Removal of the t-butoxycarbonyl, e.g. with trifluoroacetic acid in methylene chloride give amines of formula

wherein R, R₀, Ri, R₃-R₅, X, Y, V and n have meaning as previously described.

Treatment of a compound of formula IVa with a compound of the formula Z-CI wherein Z has meaning as defined for intermediates of formula XVIII, under conditions described for the preparation of intermediates of formula XIX, gives the corresponding compounds of formula IV. This method for the preparation of compounds of the invention is most appropriate for compounds of formula IV wherein X is CR2 and furthermore wherein Y is CH.

Compounds of formula IV wherein Z is heteroaryl-lower alkyl can be prepared similarly to previously described methods using e.g. an heteroaryl-alkyl halide, such as an heteroaryl- alkyl bromide. Compounds of the invention of formula IV wherein V is a direct bond are prepared from either an intermediate of formula IVa or XVI wherein V is a direct bond according to processes previously described.

The starting materials of formula XVII wherein V is a direct bond can be prepared starting from the corresponding nitro-substituted (aryl or heteroaryl)-alkyl-carboxylic acids esters or alcohols according to methods well known in the art for the conversion of esters and alcohols to amines and substituted amines.

The starting materials of formula IVa may also be similarly prepared starting e.g. from the corresponding carboxylic acid derivatives which are prepared by acylation of an appropriate amino-substituted (aryl or heteroaryl)-alkylcarboxylic acid ester with an acid of e.g. formula VII. The starting materials of formula IVa wherein V is a direct bond may also be prepared by deprotection of an intermediate of formula XVI wherein Z is t-butoxycarbonyl amino and V is a direct bond. Such intermediate of formula XVI may be prepared by condensation of a nitro-substituted aryl or heteroaryl bromide with e.g. 3-(t-butoxycarbonylamino)-propyne in the presence of e.g. Cμl, Pd (PPh₃)CI₂ and triethylamine, followed by catalytic hydrogenation.

Compounds of the invention may be converted to other compounds of the invention. For example a compound of formula IV wherein R₀ is hydrogen and Z is -SO₂ R_θ may be converted to the corresponding compound wherein R₀ is methyl, by treatment with e.g. methyl iodide in the presence of sodium hydride in dimethylformamide.

In starting compounds and intermediates which are converted to the compounds of the invention in a manner described herein, functional groups present, such as amino, carboxyl, and hydroxy groups, are optionally protected by conventional protecting groups that are common in preparative organic chemistry. Protected amino, carboxyl, and hydroxy groups are those that can be converted under mild conditions into free amino, carboxyl and hydroxy groups without the molecular framework being destroyed or other undesired side reactions taking place.

The purpose of introducing protecting groups is to protect the functional groups from undesired reactions with reaction components under the conditions used for carrying out a desired chemical transformation. The need and choice of protecting groups for a particular reaction is known to those skilled in the art and depends on the nature of the functional group to be protected (hydroxy group, amino group, etc.), the structure and stability of the molecule of which the substituent is a part and the reaction conditions.

Well-known protecting groups that meet these conditions and their introduction and removal are described, for example, in J.F.W. McOmie. "Protective Groups in Organic Chemistry", Plenum Press, London, New York, 1973, T. W. Greene, "Protective Groups in Organic Synthesis", third edition, Wiley, New York, 1999.

In the processes cited herein, reactive functional derivatives of carboxylic acids represent, for example, anhydrides (especially mixed anhydrides), acid halides, acid azides, lower alkyl esters, and activated esters thereof. Mixed anhydrides are preferably such from pivalic acid, or a lower alkyl (ethyl, isobutyl) hemiester of carbonic acid; acid halides are for example chlorides or bromides; activated esters for example succinimido, phthalimido or 4- nitrophenyl esters; lower alkyl esters are for example the methyl or ethyl esters.

Depending on the choice of starting materials and methods, the new compounds may be in the form of one of the possible isomers or mixtures thereof, for example, as substantially pure geometric (cis or trans) isomers, optical isomers (antipodes), racemates, or mixtures thereof. The aforesaid possible isomers or mixtures thereof are within the purview of this invention.

Any resulting mixtures of isomers can be separated on the basis of the physico-chemical differences of the constituents, into the pure geometric or optical isomers, diastereoisomers, racemates, for example by chromatography and/or fractional crystallization, or resolved by enzymatic resolution.

Any resulting racemates of intermediates can be resolved into the optical antipodes by known methods, e.g., by separation of the diastereoisomeric salts thereof, obtained with an optically active acid or base, and liberating the optically active acidic or basic compound. The amine intermediates can thus be resolved into their optical antipodes e.g., by fractional crystallization of salts of d- or l-carboxylic acids (e.g., d-or l-tartaric acid or d- or I- camphorsulfonic acid). Racemic products can also be resolved by chiral chromatography, e.g., high-pressure liquid chromatography using a chiral absorbent.

Finally, compounds of the invention are either obtained in the free form, or as a salt thereof if salt forming groups are present.

Compounds of the invention having basic groups can be converted into acid addition salts,

-especialJy-pharmaceutically^cceptaW inorganic acids, such as mineral acids, for example sulfuric acid, a phosphoric or hydrohalic acid, or with organic carboxylic acids, such as (Cι-C₄)-alkanecarboxylic acids which, for example, are unsubstituted or substituted by halogen, for example acetic acid, such as saturated or unsaturated dicarboxylic acids, for example oxalic, succinic, maleic or fumaric acid, such as hydroxycarboxylic acids, for example glycolic, lactic, malic, tartaric or citric acid, such as amino acids, for example aspartic or glutamic acid, or with organic sulfonic acids, such as (CrC₄)-alkylsulfonic acids (for example methanesulfonic acid) or arylsulfonic acids which are unsubstituted or substituted (for example by halogen). Preferred are salts formed with hydrochloric acid, methanesulfonic acid and maleic acid.

In view of the close relationship between the free compounds and the compounds in the form of their salts, whenever a compound is referred to in this context, a corresponding salt is also intended, provided such is possible or appropriate under the circumstances.

The compounds, including their salts, can also be obtained in the form of their hydrates, or include other solvents used for their crystallization.

The pharmaceutical compositions according to the invention are those suitable for enteral, such as oral or rectal, transdermal and parenteral administration to mammals, including man, to inhibit microsomal triglyceride transfer protein (MTP) and apolipoprotein B (apo B) secretion, and for the treatment of disorders responsive thereto, comprising an effective amount of a pharmacologically active compound of the invention, alone or in combination, with one or more pharmaceutically acceptable carriers.

The pharmacologically active compounds of the invention are useful in the manufacture of pharmaceutical compositions comprising an effective amount thereof in conjunction or admixture with excipients or carriers.especially suitable for either enteral or parenteral application. Preferred are tablets and gelatin capsules comprising the active ingredient together with a) diluents, e.g., lactose, dextrose, sucrose, mannitol, sorbitol, cellulose and/or glycine; b) lubricants, e.g., silica, talcum, stearic acid, its magnesium or calcium salt and/or polyethyleneglycol; for tablets also c) binders e.g., magnesium aluminum silicate, starch paste, gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose and or polyvinylpyrrolidone; if desired d) disintegrants, e.g., starches, agar, alginic acid or its sodium salt, or effervescent mixtures; and/or e) absorbants, colorants, flavors and sweeteners. Injectable compositions are preferably aqueous isotonic solutions or suspensions, and suppositories are advantageously prepared from fatty emulsions or suspensions. Said compositions may be sterilized and/or contain adjuvants, such as preserving, stabilizing, wetting or emulsifying agents, solution promoters, salts for regulating the osmotic pressure and/or buffers. In addition, they may also contain other therapeutically valuable substances. Said compositions are prepared according to conventional mixing, granulating or coating methods, respectively, and contain about 0.1 to75%, preferably about 1 to 50%, of the active ingredient. Suitable formulations for transdermal application include an effective amount of a compound of the invention with carrier. Advantageous carriers include absorbable pharmacologically acceptable solvents to assist passage through the skin of the host. Characteristically, transdermal devices are in the form of a bandage comprising a backing member, a reservoir containing the compound optionally with carriers, optionally a rate controlling barrier to deliver the compound of the skin of the host at a controlled and predetermined rate over a prolonged period of time, and means to secure the device to the skin.

Suitable formulations for topical application, e.g., to the skin and eyes, are preferably aqueous solutions, ointments, creams or gels well known in the art.

The pharmaceutical formulations contain an effective therapeutic amount of a compound of the invention as defined above, either alone or in combination with another therapeutic agent, at an effective therapeutic dose as reported in the art. Such therapeutic agents are well known in the art.

In conjunction with another active ingredient, a compound of the invention may be administered either simultaneously, before or after the other active ingredient, either separately by the same or different route of administration or together in the same pharmaceutical formulation.

The dosage of active compound administered is dependent on the species of warmblooded animal (mammal), the body weight, age and individual condition, and on the form of administration. A unit dosage for oral administration to a mammal of about 50 to 70 kg -may-contain between-about 1 and 1000 mg^advantageously between-about 5 and 500 mg- of the active ingredient, preferably between about 10 and 100 mg.

The present invention also relates to methods of using the compounds of the invention and their pharmaceutically acceptable salts, or pharmaceutical compositions thereof, in mammals for the treatment of elevated levels of MTP and of apo B and conditions related thereto.

The compounds of the invention are inhibitors of microsomal triglyceride transfer protein (MTP) and of apolipoprotein B (apo B) secretion and are thus useful for lowering serum lipid levels, including serum triglyceride and serum cholesterol levels. Such compounds are therefore useful for the treatment and prevention of hyperlipidemia, hypercholesterolemia and hypertriglyceridemia and diseases associated therewith, e.g., cardiovascular diseases including cardiac ischemia, atherosclerosis and its clinical sequelae, as well as obesity, pancreatitis and diabetes.

The above-cited properties are demonstrable in vitro and in vivo tests using advantageously mammals, e.g., rats, hamsters, mice, dogs, monkeys, and isolated cells or enzyme preparations. Said compounds can be applied in vitro in the form of solutions, e.g., aqueous solutions, and in vivo advantageously orally, topically or parenterally, e.g., intravenously. The dosage in vitro may range from about 10^'5 to 10^'9 molar concentrations. The dosage in vivo may range, depending on the route of administration, between about 1 and 100 mg/kg. The tests are generally known in the art. For in vivo evaluation, the compounds are generally administered as a solution or suspension, e.g., as a suspension in 3% cornstarch.

The activity of a compound according to the invention can be assessed by the following methods:

The inhibition of the cellular secretion of apo B is determined as follows:

Hep G2 cells are maintained in T-75 culture flasks (Corning) in Dulbecco's modified Eagles

Medium (DMEM; Gibco-BRL) supplemented with 10% fetal calf serum Gibco-BRL) in a humidified^nrros rrere^on niιιg-5yό^^ compounds, Hep G2 cells from the T-75 maintenance flasks are harvested and seeded in

96-well culture plates (Corning) and are grown for 72 hours (approximately 80% confluent).

Test compound is dissolved at 1 mg/mL (w/v;1-5 mM) in dimethyl sulfoxide DMSO; Sigma) as stock solution. Prior to use, the stock solution of compound is diluted to 133 μM with DMSO and diluted further with growth medium (DMEM containing 10% fetal calf serum) to obtain 1 μM of compound in 100 μl of growth medium. 100 μL of growth medium containing the test compound is added to separate wells of a 96-well culture plate containing Hep G2 cells. For testing dose response of compound, a stock solution of test compound in DMSO is made at 665 μM and various dilutions from this solution are made in growth medium to obtain range of concentration of compound from 0.01 μM to 5 μM in 100 μL of growth medium. 100 μL of the growth medium containing different concentrations of test compound is added to separate wells containing Hep G2 cells. Twenty-four hours later, growth medium is collected and assayed by specific ELISA for apolipoprotein B (apo B). At the same time Hep G2 cells from wells are assayed for protein (BioRad ; cat. # 500-0006 ) and/ or cell viability (Promega; CellTiter 96 Aqueous, cat. # G3581 ). Inhibitors are identified as compounds that decrease apo B secretion into the medium without decreasing the total cellular protein and/or cell viability. For performing apo B ELISA, an antisera for human apo B is made by immunizing rabbit with purified human apo B. The antisera is further purified by using an affinity column (CNBr activated Sepharose 4B, Pharmacia) with human LDL as ligand and used as primary antibody for human apo B. A secondary antibody for apo B is prepared by conjugating the human apo B antibody with alkaline phosphatase (Sigma). The ELISA for apo B is performed as follows. 15 μL of primary antibody solution prepared against apo B is diluted to a final volume of 10 mL with coating buffer (containing 15 mM sodium carbonate, 35 mM sodium bicarbonate, 3 mM sodium azide, pH 9.6). 200 μL of diluted antibody solution is added to each well of a 96 well plate (Maxisorb, Nunc , cat. # 439454). After an overnight incubation at 4^°C, the antibody solution is removed. Nonspecific sites on the plastic well are blocked by adding 300 μL of blocking solution containing phosphate buffered saline (PBS) , 1 % (w/v) bovine serum albumin (Sigma), pH 7.4) and incubated for 45 minutes at room temperature. After removing blocking solution, 200 μL of dilution buffer (containing PBS/ 0.05% Tween 20 / 5 mM decyl sodium sulfate (Acros Organics) / 2% BSA, pH 7.4) containing 20 μL of growth medium from Hep G2 cells or 1 - 30 ng of apo B standards (prepared in dilution buffer) is added to each well. After 2 hours incubation at 37^°C, solution from each well is removed and washed five times with washing buffer (containing PBS and 0.05% Tween 20, pH 7.4). 200 μL of diluted conjugated secondary antibody for apo B (15 μL diluted to a final volume of 10 mL in dilution buffer) is added to each well. After 2 hours incubation at 37°C, the solution is removed and the well is washed five times with washing buffer. p-Nitrophenyl phosphate disodium hexahydrate solution (Sigma, cat. # 104-0) is prepared in substrate buffer (containing 0.95M diethanolamine / 0.5mM MgCI2 / 3 mM sodium azide, pH 9.5) at a concentration of 1 mg/mL and 200 μL of substrate solution is added to each well and incubated for 45-60 minutes. Absorbance of each well is read at 405 nm using a Beckman Biomek workstation. Apo B concentration is calculated from a standard curve generated from purified LDL standards that are run in parallel in the same assay. Secreted apo B values are normalized with the total cellular protein assay and/or cell viability assay.

The inhibition of MTP is measured as follows:

Inhibition of the lipid transfer activity of MTP can be quantitated by measuring the inhibition of transfer of radiolabeled triglyceride from donor vesicles to acceptor vesicles in presence of soluble rat MTP. The procedure for preparing MTP is based on the method of Wetterau and Zilversmit (Biochim. Biophys. Acta (1986) 875:610). Briefly rats are decapitated under ether anesthesia. The liver is placed in ice cold sucrose buffer (contains 0.25M sucrose, 50 mM Tris HCI, 1 mM EDTA, 0.02% sodium azide, pH 7.4) rinsed several times with the sucrose buffer.

All subsequent steps are performed on ice. A 57% homogenate (120 g/210 mL) of rat liver in 0.25M sucrose buffer is prepared by using a Potter-Elvehjem homogenizer. The homogenate is then centrif uged at 4°C for 30 min at 13,000 x g to remove large cellular organells. The supernatant is then centrifuged for 90 min at 105,000 x g to pellet the microsomes. The pellet is resuspended in 10 mM Tris-HCI buffer pH 8.6. and centrifuged for 90 min at 105,000 x g. The washed pellet is then resuspended in 1 mM Tris buffer (pH 8.6) and centrifuged for 2 hrs. The pellet is resuspended in 28.5 mL of 0.25M sucrose solution and 1 mL aliquotes containing 4.2 g of liver are stored frozen at -80^°C until needed. Prior to performing the assay, the thawed pellet is suspended in 12 mL of cold Tris-HCI, 50 mM KCI, 5 mM MgCI, pH 7.4 and 1.2 mL of a 0.54% deoxycholate solution (pH 7.4) is added^lowly-with^entle-mixing_^The^uspension-is-kept^n ice-for-30-inin-and hen centrifuged at 105,000 g for 75 min. The supernatant containing soluble MTP is dialyzed against assay buffer (150 mM Tris-HCI, 40 mM NaCI, 1 mM EDTA, 0.02% NaN₃, pH 7.4). The protein content is measured using the Sigma Lowry micro total protein method and reagents (Sigma Cat. # 690A). The rat MTP is diluted with assay buffer to contain 15 μg protein per 50 μL and stored at 4°C.

Donor and acceptor liposomes are prepared as follows. For preparation of donor vesicles, 12.4 mg of egg phosphatidylcholine (Sigma, cat. # P-3556), 5.2 mgs of cardiolipin (Sigma, Cat. # C-0563) and 8 mg of hydroxybutylate toluene are dissolved in 4 mL of chloroform. To this solution, 34.8 μL of ³H labeled Triolein (Amersham, Cat. # TRA 191 , glycerol tri[1 ,9- ³H]oleate) is added and mixed. 200 μL of this mixture is transferred into a screw cap glass vial, dried under nitrogen and reconstituted in 2 mL of assay buffer. The lipid suspension is sonicated for 30 min at 1.5 setting with pulse at 75 using Branson 450 sonifier in a water bath with ice. For preparation of acceptor vesicles, 18 mgs of egg phophatidylcholine and 4 mgs of hydroxybutylated toluene is added in 1 mL of chloroform. A 200 μL aliquot from this mixture is transferred into a screw cap glass vial. To this vial, 10 μL of Triolein ( 0.92 mg/ml in chloroform) and 2 μL of ¹ C labeled phosphotidylcholine (Amersham, cat. # CFA 695, L-3- phosphatidylcholine,1,2-di[1-¹⁴C]oleoyl) are added and dried under nitrogen and reconstituted in 2 ml of assay buffer. The lipid suspension is sonicated using Branson 450 sonifier as described above. The donor and acceptor liposomes are centrifuged for 2 hours at 7 ° C at 46,000 rpm in Ti50 rotor using Beckman Ultracentrifuge. The upper 75% of the supernatant is carefully removed and stored at 4°C until used for MTP transfer assay .

MTP activity is measured using a MTP transfer assay. In this assay, donor and acceptor vesicles are mixed together with soluble MTP and test compound to measure the transfer of triglycerides from donor vesicles to acceptor vesicles. 50 μL of donor vesicles, 50 μL of acceptor vesicles, 20 μL of bovine serum albumin (10% w/v) and 50 μL of MTP (15 μg protein) are added along with various concentrations of test compound in a final volume 450 μL of assay buffer.

After incubation at 37 °C for 45 min, the triglyceride transfer is terminated by addition of 300 μL of DEAE cellulose suspension (50%, w/v). After 4 min of vortexing, the donor vesicles bound to the DEAE cellulose are separated from acceptor vesicles by centrifuging at 14,000 rpm for 7 min.250 μL of supernatant containing acceptor vesicles are counted using 5.5 mL of Ready safe scintillation solution (Beckman, cat. # 158735). The ¹⁴C and ³H counts are used to calculate the percent recovery of acceptor liposomes and the percent of triglyceride transfer using first order kinetics. Inhibition of triglyceride transfer by test compound is calculated by measuring the decrease in ³H label of triglyceride present in the acceptor vesicles as compared to controls where no test compound is present.

Illustrative of the invention, the compound of Example 1 demonstrates an IC₅₀ of about 1nM in the apo B assay and an IC₅₀ of about 90 nM in the MTP assay.

The in vivo serum triglyceride lowering effect of the compounds of the invention can be determined by measuring their effect on triglyceride levels in mice, rats or dogs according to methodology well known in the art, e.g., in a model of pre-established hypertriglyceridemia in fructose fed rats or in normolipidemic rats.

The in vivo serum cholesterol lowering effect of the compounds of the invention can be determined by measuring their effect on cholesterol levels in mice, rats, or dogs according to methodology well known in the art, e.g., in normolipidemic rats.

Illustrative of the invention, the compound of example 1 lowers both plasma triglycerides and cholesterol at a dose of 5 mg/kg. p.o. in the rat.

The following examples are intended to illustrate the invention and are not to be construed as being limitations thereon. Temperatures are given in degrees Centrigrade. If not mentioned otherwise, all evaporations are performed under reduced pressure, preferably between about 15 and 100mm Hg (= 20-133 mbar). The structure of final products, intermediates and starting materials is confirmed by standard analytical methods, e.g., microanalysis and spectroscopic characteristics (e.g., MS, IR, NMR). Abbreviations used are those conventional in the art. The concentration for [α] determinations is expressed in mg/mL. Compounds are purified by standard methods, e.g., recrystallization or flash chromatography.

Abbreviations for reagents, solvents and the like are those conventionally used in the art. Hlustra1rve^lτeτeOf-aτeihe^o1lowirrgr

BOC: t-Butoxycarbonyl DEAD: Diethyl azodicarboxylate DIEA: Diisopropylethyl amine DMF: Dimethylformamide

DMSO: Dimethyl sulfoxide

EDCI: 1 -Ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride

HBTU: O-Benzotriazolyl-N,N,N',N'-tetramethyluronium hexafluorophosphate

HOAt: 1 -Hydroxy-7-azabenzotriazole

MTBE: Methyl t-butyl ether

THF: Tetrahydrofuran

EXAMPLES

A. Preparation of 6-methyl-4'-trifluoromethyl-1.1'-biphenyl-2-carboxylic acid

A solution of 2-bromo-3-methylbenzoic acid (21.5 g, 100 mmol) in 500 mL of methanol and 8 mL of concentrated sulfuric acid is refluxed overnight. Methanol is removed under reduced pressure, the residue is taken up in ether, washed with sodium bicarbonate, brine, and dried over magnesium sulfate, filtered, and evaporated under reduced pressure to give methyl 2-bromo-3-methylbenzoate as an oil.

A mixture of methyl 2-bromo-3-methylbenzoate (22.33 g, 97.5 mmol), potassium phosphate (82.8 g, 390 mmol), [1 ,1'-bis(diphenylphosphino)-ferrocene]dichloro palladium (II) complex with dichloromethane (1:1) (3.98 g, 4.87 mmol), and p-trifluoromethylphenylboronic acid (22.2 g, 117 mmol) in 500 mL of DME is degassed and refluxed under an atmosphere of argon overnight. The mixture is concentrated, poured into water and extracted with ethyl acetate. The combined organic extracts are washed with brine, dried over magnesium sulphate, filtered and evaporated under reduced pressure. The residue is purified by silica gel chromatography eluting with ethyl acetate / toluene (1 :9) to give methyl 6-methyl-4'- trifluoromethyl-1 ,1 '-biphenyl-2-carboxylate.

AlteiΩative.y_τ-metøyI^-methv1-44^ as follows:

To a slurry of 3-methylsalicylic acid (200 g) in 600 mL of methanol at -15⁹ is added 65.7 g of concentrated sulfuric acid. The mixture is treated at reflux temperature for 5 days. The reaction mixture is concentrated, and methyl-t-butyl ether (500 mL) and water (250 mL) are added. The ether layer is separated, washed with bicarbonate solution and evaporated to dryness to give methyl 3-methylsalicylate an oil.

A mixture of methyl 3-methylsalicylate (150 g), pyridine (178.5 g) and methylene chloride (1500 mL) is cooled to -5^s. Triflic anhydride (305.6 g) is added over 30 minutes. 4-Dimethylaminopyridine (3.31 g) is then added, the reaction mixture is stirred at room temperature overnight, washed with 1 N HCI, then with saturated sodium bicarbonate solution and finally with brine. The solution is dried over magnesium sulfate, and evaporated to dryness in the presence of toluene. The residual oil is dissolved in toluene to obtain a volume of 3000 mL and the solution of methyl 2-trifluoromethanesulfonyloxy-3- methylbenzoate is used as is in the next step.

A solution of p-trifluoromethylbromobenzene (814.8 g) and triisopropoxyborane (681.0 g) in tetrahydrofuran (6300 mL) is cooled to -78⁹ and n-butyllithium (2.5 m in hexanes, 1448 mL) is added over 30 minutes at a temperature below -60² to yield p- trifluoromethylphenylboronic acid.

To a solution of methyl 2-trifluoromethanesulfonyloxy-3-methylbenzoate (900 g, in toluene), potassium carbonate (629.6 g), tetrahydrofuran (2700 mL) and deionized water (5400 mL) under nitrogen is added tetrakis (triphenylphosphine) palladium (0) (104.6 g). To this is added the above solution of p-trifluoromethylphenylboronic acid and the mixture is heated at reflux for 2 days. The reaction mixture is filtered and evaporated to dryness. The residue is partitioned between water and ethyl acetate. The ethyl acetate layer is separated and evaporated to dryness. The residue is taken up in heptane-ethyl acetate (9:1), the mixture is filtered and the filtrate is evaporated to dryness to give methyl 6-methyl-4'- trifluoromethyl-1,1'-biphenyl-2-carboxylate as an oil.

A mixture of methyl 6-methyl-4'-trifluoromethyl-1,1'-biphenyl-2-carboxylate (13.6 g, 46.3 mmoL)_r-and_lN-NaQH_(9P.5 ml , 92.5 mmol) in 295 ml of ethanol is refluxed for 5 h. Water is added to the mixture and the aqueous layer is washed with ether. The aqueous layer is acidified with 1 N HCI and extracted with ethyl acetate, dried over magnesium sulfate, filtered, and concentrated under reduced pressure. The product is purified by crystallization from ethyl acetate / hexanes to yield 6-methyl-4'-trifluoromethyl-1 ,1'-biphenyl-2-carboxylic acid melting at 202-203°C. ¹HNMR CDCI₃: 300 MHz): δ 7.87 (1H, d),7.65(2H, d), 7.47(1 H, d),7.35 (1H, t), 7.25 (2H, d), 2.05(3H, s). MS m/z 279 (M -1).

Alternately, 6-methyl-4'-trifluoromethyl-1,1'-biphenyl-2-carboxylic acid can be prepared as follows:

3-Methylsalicylic acid is esterified to methyl 3-methylsalicylate by reaction under reflux for about 48 hours with methanol in the presence of trimethyl orthoformate (4.0 moles) and concentrated sulfuric acid (1.1 moles) while removing by distillation the generated methyl formate and replacing the methanol which is lost by distillation. The reaction mixture is then evaporated to dryness at 40⁹ under vacuum and toluene is added. The toluene solution is washed with water, then 20% aqueous potassium bicarbonate solution and saturated sodium chloride solution. The toluene solution is filtered through neutral activated aluminum oxide and evaporated to dryness to yield methyl 3-methylsalicylate as a liquid.

To a solution of 1.0 mole of methyl 3-methylsalicylate in toluene are added 2.33 moles of 4- methylmorpholine and 0.022 moles of 4-dimethylaminopyridine. The resulting solution is then treated with 1.07 moles of trifluoromethanesulfonic acid anhydride (triflic anhydride) at -15⁹C. The reaction mixture is stirred overnight at -3 to -4⁹C and washed consecutively with 3.7% aqueous HCI, 20% potassium bicarbonate solution and saturated sodium chloride solution. The toluene solution is then filtered through alumina and evaporated to dryness at less than 50⁹C and about 40 mm Hg pressure. The residue is distilled at 50^gC and 5 mmHg pressure to obtain methyl 3-methyl-2-trifluoromethanesulfonyloxybenzoate as an oil. A solution of 140.75 g of 4-bromobenzotrifluoride and 117.6 g of triisopropylborate in 1050 mL of dry and peroxide-free THF is cooled to -72± 3⁹C. 275 mL of 2.5 M n-BuLi solution in hexane are slowly added over a period of 90 minutes at such a rate that the internal temperature of the reaction mixture remains below -60⁹C. The mixture is stirred at -65 to - 70^SC for 30 minutes after the addition and warmed to 0⁹C over 90 minutes. This solution of diisoprop-yj--i--trjfιuo-oj- eir^^

A solution of 155 g of methyl 3-methyl-2-trifluoromethylmethanesulfonyloxybenzoate, 107.8 g of potassium carbonate in 467.5 mL of THF and 935 g of deionized water is stirred under nitrogen for 20 minutes. 15.0 g of Tetrakis(triphenylphosphine)palladium (0) and -1610 mL of boronate solution from step above are added and the reaction mixture is heated under gentle reflux (~64⁹C) for 16 hours. The reaction mixture is cooled to room temperature and filtered through a pad of 40 g of the filter agent Celite^® 521. The filter cake is washed with 100 mL of THF and the total filtrate is partially evaporated under reduced pressure (110-120 mbar/40⁹C) to remove about 1500 mL of distillate and obtain a three phase mixture (about 1250 mL). 500 mL of f-Butyl methyl ether and 200 mL of 2% sodium chloride solution are added. The mixture is stirred for 5 minutes and filtered through a pad of 40 g of Filter agent, Celite^® 521 and the filter cake is washed with 100 mL of f-butyl methyl ether. The top organic layer (1100 mL) is separated from the bottom aqueous layer (950 mL). The bottom aqueous layer (950 mL) is extracted with 300 mL of f-butyl methyl ether. The combined upper organic phases are washed with 300 mL of 2% sodium chloride solution and evaporated under reduced pressure (28 mbar/40⁹C) to obtain methyl 6-methyl-4'- trifluoromethyl-1 ,1 '-biphenyl-2-carboxylate.

6N Sodium hydroxide (260 mL) is added slowly at room temperature to a solution of 183 g of methyl 6-methyl-4'-trifluoromethyl-biphenyl-2-carboxylate in 1200 mL of methanol. The reaction mixture is heated under gentle reflux for 2.5 hours, cooled to room temperature and diluted with 300 mL of water. The reaction mixture is evaporated under reduced pressure (110-120 mbar/40⁹C) to a suspension of about 750 mL which is filtered through the filter agent Celite^® 521 , and the filter cake is washed sequentially with 250 mL of water and 250 mL of heptane. The organic layer is separated and the aqueous layer is washed with 250 mL of heptane. The aqueous layer is acidified with 500 mL of 4N hydrochloric acid and extracted with ethyl acetate. The ethyl acetate extract is washed with water and filtered through the filter agent Celite^® 521. The ethyl acetate solution (ca.1000 mL) is evaporated under reduced pressure (110-120 mbar/40⁹C) to a volume of 600 mL to which is added heptane (3125 mL). The suspension is then heated to reflux until a clear solution is obtained, the solution is cooled to 0⁹C and the resulting solid is filtered off to yield 6-methyl- 4'-trifluoromethyl-1 ,1'-biphenyl-2-carboxylic acid.

B. Preparation of 2-(4-trifluoromethyl-phenyl)-nicotinic acid chloride

To a solution of 2-chloronicotinic acid (2.00 g, 12.69 mmol) in DMF (40 mL) at 0⁹C is added cesium carbonate (4.96 g, 15.23 mmol) followed by iodomethane (0.95 mL, 15.23 mmol). The reaction mixture is warmed to room temperature and stirred 16 h. Dilution with ethyl acetate is followed by washing with water, 8% NaHCO₃ solution and brine. The organic layer is dried (MgSO4₄) and concentrated under reduced pressure to give 2-chloronicotinic acid methyl ester as an oil: ¹H NMR (CDCI₃, 300 MHz): δ 8.55 (1H, dd), 8.18 (1H, dd), 7.33 (1H, dd), 3.97 (3H, s).

2-(4-Trifluoromethyl-phenyl)-nicotinic acid methyl ester is prepared by palladium [PdCI₂ (dppf)] catalyzed coupling of 2-chloronicotinic acid methyl ester (1.45 g, 8.45 mmol) and 4- trifluoromethylbenzeneboronic acid (2.41 g, 12.68 mmol) to give an oil: ¹H NMR (CDCI₃, 300 MHz): δ 8.80 (1H, dd), 8.19 (1H, dd), 7.67 (4H, q), 7.40 (1H, dd), 3.70 (3H, s). MS (ES+) m/z 282 (M+1).

To a solution of 2-(4-trifluoromethyl-phenyl)-nicotinic acid methyl ester (0.626 g, 2.228 mmol) in 1 :1 THF:H₂O (10 mL) is added LiOH H₂O (0.187 g, 4.456 mmol). After 3.5 h, the reaction mixture is concentrated to dryness under reduced pressure. To a slurry of the crude lithium salt in methylene chloride (10 mL) is added oxalyl chloride (0.78 mL, 8.91 mmol) followed by a few drops DMF. After stirring 16 h, the reaction mixture is concentrated to dryness under reduced pressure to give the acid chloride which is used as is without purification.

C. Preparation of 2-(4-trifluoromethylbenzyloxy)-3-rnethylbenzoic acid

To a solution of 2-hydroxy-3-methylbenzoic acid (18.1 g, 119 mmol) in 70 mL of methanol is added dropwise 3.3 mL of concentrated sulfuric acid. The resulting solution is heated under reflux for 24 hours. Another 7 mL of concentrated sulfuric acid is added and the reaction is heated at 70⁹C overnight. The mixture is cooled and a pink layer separated. This layer is dissolved in 200 mL of ethyl acetate and the solution is washed with saturated aqueous sodium bicarbonate and brine, dried over sodium sulfate, filtered and concentrated to give an oil which is purified by chromatography on silica gel using a mixture of 30% ethyl acetate and 70% hexane as the eluent. Methyl 2-hydroxy-3-methylbenzoate is obtained as a clear oil.

Sodium hydride is added to a solution of methyl 2-hydroxy-3-methylbenzoate (694 mg, 4.18 mmol) in 15 mL of DMF at 0⁹C. The reaction is stirred for 15 minutes after which 4- trifluoromethyl-benzyl bromide (1.0 g, 4.18 mmol) is added. The reaction is allowed to warm up to room temperature and stirred overnight. The mixture is partitioned between ethyl acetate and water. The organic phase is washed with brine, dried over sodium sulfate, filtered and concentrated. The residue is chromatographed on silica gel using 4% ethyl acetate in hexane as the eluent to obtain methyl 2-(4-trifluoromethylbenzyloxy)-3- methylbenzoate as a clear liquid.

To a solution of methyl 2-(4-trifluoromethylbenzyloxy)-3-methylbenzoate (1.17g, 3.61 mmol) is added 10 mL of 1 N LiOH. The mixture is stirred at room temperature for 3 days, then concentrated under vacuum. The residue is washed with hexane, then acidified to pH 1 with 6N HCI. A white precipitate forms which is extracted with ethyl acetate. The organic solution is dried over sodium sulfate, filtered and concentrated to give the title acid as a white solid , m.p. 104-106⁹C.

Example 1

β-Methyl-Λ/-r4-rr2-(2-pyridinvnethvπaminolphenvπ-4'-ftrifluoromethvn-r .1'-biphenvπ-2- carboxamide

Step A. Λ/-(4-Nitrophenyl)-2-pyridineethanamine

4-chloronitrobenzene (4.13 g, 30 mmol) and 2-(2-aminoethyl)pyridine (7.33g, 60 mmol) are heated at 120⁹ for 16 hr in DMSO (7 mL). The reaction mixture is then cooled and partitioned between methylene chloride (150 mL) and water (250 mL). The organic phase is washed with an additional portion of water, dried over sodium sulfate, and applied to a silica gel column (120 g of silica gel). The column is eluted with methylene chloride to give product as a yellow solid; MS: 244.0 (M+H).

Step B. Λ/¹-[2-(2-Pyridinyl)ethyl]-1,4-benzenediamine

A mixture of the product from Step A (1.98 g, 8.14 mmol) and 10% Pd/C (0.47g) in ethanol (80 mL) is shaken under 40 psi of hydrogen for 3 hours. The solution is filtered and concentrated under vacuum to give desired product as a pink solid. Step C. 6- ethyl-Λ^-[4-[[2-(2-pyridinyl)ethyl]amino]phenyl]-4'-(trif luoromethyl)-[1 ,1 '- biphenyl]-2-carboxamide

6-Methyl-4'-trifluoromethyl-1 ,1'-biphenyl-2-carboxylic acid (2.688g, 9.59 mmol) and 1- hydroxy-7-azabenzotriazole (HOAt) (1.43 g, 10.55 mmol) are dissolved in DMF (15 mL), and EDCI (2.02g, 10.55 mmol) is added. After stirring for 1h, a solution of the amine from Step B (2.05g, 9.59 mmol) in DMF is added and the reaction is stirred for an additional 16 hours at rt. Saturated aqueous sodium bicarbonate (10 mL) is added slowly over 30 minutes, and then an additional portion of water (50 mL) is added over 15 min. The mixture is stirred and filtered to give a gray solid which is washed with water (2 x 10 mL). The solid is then dissolved in methylene chloride (40 mL), and residual suspended water is removed. The methylene chloride solution is applied to a silica gel column (120 g of silica gel) and eluted first with methylene chloride (6 x 250 mL fractions), then 1 % methanol in methylene chloride (4 x 250 mL fractions), 1.5% methanol in methylene chloride (4 x 250 mL fractions), 3% methanol in methylene chloride (4 x 250 mL fractions), and 5% methanol in methylene chloride (4 x 250 mL fractions) to give derived product as a white solid. This material is triturated with MTBE, dissolved in methylene chloride and the solution is treated with charcoal and evaporated to dryness. The residue is triturated again with MTBE to give the title amide, m.p. 144-145 ⁹C.

Example 2

6-Methyl-/v-r4-r2-(2-pyridinyloxy)ethyllphenvn-4'-(trifluoromethyl.-n.1'-biphenvn-2- carboxamide

Step A. 2-[2-(4-Nitrophenyl)ethoxy]-pyridine

To a solution of triphenylphosphine (2.07 g, 7.89 mmol) in 20 mL of THF at room temperature is added diethyl azodicarboxylate (DEAD), (1.37 g, 7.89 mmol) dropwise with stirring. After 30 minutes, 2-(4-nitrophenyl) ethyl alcohol and 2-hydroxypyridine are added separately. The mixture is stirred at room temperature for 24 hours. The mixture is then concentrated, the residue is dissolved in methylene chloride and chromatographed on a silica gel column using 500 mL of hexane-ethyl acetate (4:1 mixture), followed by 1 L of hexane-ethyl acetate (2:3 mixture) as eluent. The desired product is isolated as an oil which crystallizes on standing. MS: 245 (M + 1).

Step B. 4-[2-(2-Pyridinyloxy)ethyl]-benzenamine

The nitro compound from Step A (480 mg, 1.97 mmol) is shaken overnight in ethanol (20 mL) with 20 mg of 10% palladium on carbon under 40 psi of hydrogen gas. The mixture is then filtered and the filtrate is concentrated to give the desired amine as a light pink solid; MS: 214.9 (M + 1).

Step C. 6-Methyl-/V-[4-[2-(2-pyridinyloxy)ethyl]phenyl]-4'-(trifluoromethyl)-[1 ,1 '- biphenyl]-2-carboxamide

A mixture of 6-methyl-4'-trifluoromethyl-1 ,1'-biphenyl-2-carboxylic acid (200 mg, 0.71 mmol), aniline (153 mg, 0.71 mmol) , EDCI (137 mg, 0.71 mmol), and HOAt (97 mg, 0.71 mmol) in DMF is stirred overnight at room temperature. The mixture is then partitioned between water and ethyl acetate. The organic solution is washed with brine, dried over sodium sulfate, and concentrated to give an oil. The residual oil so obtained is chromatographed on silica gel using hexane-ethyl acetate (7:3 mixture) as the eluent to obtain the title compound; MS: 477 (M + 1); mp. 111-114⁹C.

Example 3

6-Methyl-.V-r4-r2-(2-pyridinyl.ethoxylphenvπ-4^,-ftrifluoromethvn-ri.1'-biphenvπ-2- carboxamide

Step A. 2-[2-(4-Nitrophenoxy)ethyl]-pyridine

To a solution of 2-(2-hydroxyethyl)pyridine (2.0 g, 16.24 mmol) in 20 mL of DMF at 0⁹C. is added sodium hydride. After 15 minutes, 1-chloro-4-nitrobenzene (1.7 g, 10.8 mmol) is added. The mixture is stirred at 0⁹C. for 1 h, then heated to 80⁹C overnight. The mixture is partitioned between ethyl acetate and water, and the organic solution is washed with brine, dried over sodium sulfate, filtered, and concentrated. The residue is chromatographed on silica gel using hexane-ethyl acetate (6:4 mixture) as the eluent to give the title compound; MS: 245 (M + 1).

Step B. 4-[2-(2-Pyridinyl)ethoxy]-benzenamine

The nitrobenzene derivative from Step A (140 mg, 0.57 mmol) is shaken under 40 psi of hydrogen gas in 20 mL of ethanol with 20 mg of 10% palladium on carbon for 6 h. The mixture is filtered and concentrated to give the title aniline as an oil; MS: 215 (M + 1). Step C. 6-Methyl-iV-[4-[2-(2-pyridinyl)ethoxy]phenyl]-4'-(trifluoromethyl)-[1 ,1 '- biphenyl]-2-carboxamide

A mixture of EDCI (108 mg, 0.56 mmol), HOAt (76 mg, 0.56 mmol), 6-methyl-4'- trifluoromethyl-1,1'-biphenyl-2-carboxylic acid (157 mg, 0.56 mmol) and the aniline from Step B (120 mg, 0.56 mmol) is stirred in DMF at room temperature overnight. The mixture is partitioned between water and ethyl acetate. The organic solution is washed with brine, dried over sodium sulfate, and concentrated. The residue is chromatographed on silica gel using hexane-ethyl acetate (1:1 mixture) as the eluent to obtain the title compound, m.p. 44- 45 ⁹C; MS: 477 (M + 1).

Example 4

Prepared similarly to the previous examples are the compounds listed below. Starting with 2-chloro-5-nitropyridine and 2-chloro(3- or 4)-methyl-5-nitropyridine instead of 4- chloronitrobenzene gives the corresponding pyridyl compounds, e.g. compounds (a)-(m) and (o)-(r).

41

-43-

Example 5

N-r4-r2-r(Phenylsulfonyl.aminolethyllphenyll-4^,-arifluoromethvn-ri.r-biphenyll-2- carboxamide

Step A. Λ/-[2-(4-Aminophenyl)ethyl]-benzenesulfonamide

^"cFξcC

Benzenesulfonyl chloride (1.77g, 10 mmol) is added dropwise to a solution of triethylamine

(2.53 g, 25 mmol) and 4-aminophenethylamine (1.36 g, 10 mmol) in 100 mL of methylene chloride at room temperature. The mixture is concentrated to remove the methylene chloride, and the residue is taken up in ethyl acetate and washed with brine, 0.1 N HCI, brine, and saturated aqueous sodium bicarbonate. The organic solution is then dried over sodium sulfate and concentrated to give the derived sulfonamide which is used without further purification.

Step B. /v-[4-[2-[(Phenylsulfonyl)amino]ethyl]phenyl]-4'-(trifluoromethyl)-[1 ,1 '- biphenyl]-2-carboxamide

A solution of 4'-(trifluoromethyl)-1 ,1'-biphenyl-2-carboxylic acid (300 mg, 1.1 mmol), the aniline from Step A (276.2 mg, 1 mmol), HOAt (408 mg, 3.0 mmol), HBTU (1.12 g, 3.0 mmol) and DIEA (388 mg, 3.0 mmol) in 5 mL of DMF is stirred at room temperature overnight. The DMF solution is diluted with 200 mL of brine and the mixture is filtered. The collected solid is taken up in 100 mL of ethyl acetate and the solution is washed with 1 N HCI, brine, saturated aqueous sodium bicarbonate, and brine, dried over sodium sulfate, and concentrated to give the crude product. The crude product is purified by chromatography on silica gel using a hexane-ethyl acetate (65:35) mixture as the eluent. to give the title amide as a white solid, m.p. 163-163.5⁹C. Example 6

r2-rr4-rrr6-Methyl-4'-(trifluoromethvn-ri.1'-biphenyll-2-yllcarbonvnaminolphenvnaminol- ethvH-carbamic acid, methyl ester

Step A. Λ.¹-(4-Nitrophenyl)-1,2-ethanediamine

Ethylenediamine (49.6 g, 827 mmol) is added to 1 -chloro-4-nitrobenzene (13 g, 82.5 mmol) and the mixture is heated to 125⁹C overnight. A yellow precipitate forms. The mixture is filtered and the solid is dried to give the desired nitro compound; MS: 180 (M - 1) and 182 (M + 1).

Step B. [2-[(4-Nitrophenyl)amino]ethyl]-carbamic acid, methyl ester

Methyl chloroformate (261 mg, 2.76 mmol) is added to a solution of the diamine derivative from Step A (500 mg, 2.76 mmol) in 10 mL of DMF containing triethylamine (559 mg, 5.52 mmol) at 0 ⁹C. The mixture is allowed to warm to room temperature and is stirred overnight after which it is partitioned between water and ethyl acetate. The organic solution is washed with brine, dried over sodium sulfate, filtered, and concentrated to give the desired compound as a yellow solid; MS: 240 (M + 1).

Step C. [2-[(4-Aminophenyl)amino]ethyl]-carbamic acid, methyl ester

A solution of the nitrobenzene derivative from Step B (600 mg, 2.51 mmol) in ethanol (20 mL) with 60 mg of 10% palladium on carbon is shaken under 40 psi of hydrogen for 6 hours. The mixture is filtered and concentrated to give desired compound as a red oil; MS:

209.9 (M + 1).

Step D. [2-[[4-[[[6-Methyl-4'-(trifluoromethyl)-[1 ,1 '-biphenyl]-2-yl]carbonyl]amino]- phenyl]amino]ethyl]-carbamic acid, methyl ester

A solution of 6-methyl-4'-trifluoromethyl-1 ,1'-biphenyl-2-carboxylic acid (200 mg, 0.71 mmol), aniline derivative from Step C (149 mg, 0.71 mmol), EDCI (137 mg, 0.71 mmol) and HOAt (97 mg, 0.71 mmol) is stirred in DMF overnight at room temperature. The mixture is partitioned between ethyl acetate and water, and the organic phase is washed with brine, dried over sodium sulfate, filtered, and concentrated to give the crude product as a black oil. This residue is chromatographed on silica gel using a hexane-ethyl acetate mixture (1 :1) as the eluent to give title compound as a yellow foam; m.p. 66-68 ⁹C; MS: 472 (M + 1) and 470 (N --1-).

Example 7 2-rr4-rr6-Methyl-4'-(trifluoromethvn-n.1^,-biphenvn-2-vncarbonvnaminolphenoxy1ethvn- carbamic acid, methyl ester

Step A. [2-(4-Nitrophenoxy)ethyl]-carbamic acid, t-butyl ester

To a solution of triphenylphosphine (1.414 g, 5.39 mmol) in 20 mL of THF is added DEAD dropwise with stirring. After 30 minutes at room temperature, 4-nitrophenol and t-butyl N-(2- hydroxyethyl)carbamate is added separately. The mixture is then stirred overnight at room temperature. The mixture is concentrated to give a yellow oil which is chromatographed on silica gel using a hexane-ethyl acetate mixture (60:40) as the eluent. Product is obtained as a light pink oil which is used directly in the next step.

Step B. [2-(4-Aminophenoxy)ethyl]-carbamic acid, t-butyl ester

The nitrobenzene derivative from Step A (590 mg) is shaken under 40 psi of hydrogen gas in ethanol with 60 mg of 10% palladium on carbon at room temperature overnight. The reaction is then filtered and concentrated to give the desired aniline derivative as an oil. MS: 253 (M + 1). Step C. 2-[[4-[[6-Methyl-4'-(trif luoromethyl)-[1 , 1 '-biphenyl]-2-yl]carbonyl]amino]- phenoxy]ethyl]-carbamic acid, t-butyl ester

A mixture of 6-methyl-4'-trifluoromethyl-1,1'-biphenyl-2-carboxylic acid (500 mg, 1.78 mmol), the carbamate from Step B (450 mg, 1.78 mmol) HOAt (243 mg, 1.78 mmol), and EDCI (342 mg, 1.78 mmol) in DMF is stirred at room temperature overnight. The mixture is partitioned between water and ethyl acetate. The organic solution is washed with brine, dried over sodium sulfate, filtered and concentrated to give an oil. The crude product is chromatographed on silica gel using a hexane- ethyl acetate mixture (70:30) as the eluent. The desired compound is obtained as a slightly pink, clear oil. MS: 515 (M + 1).

Step D. Λ/-[4-(2-Aminoethoxy)phenyl]-6-methyl-4'-(trifluoromethyl)-[1 ,1 '-biphenyl]-2- carboxamide

Trifluoroacetic acid (1.55 g, 13.6 mmol) is added to a solution of the BOC derivative from Step C (700 mg, 1.36 mmol) in methylene chloride at 0⁹C. The mixture is allowed to warm to room temperature and stirred overnight after which it is concentrated. The residue is partitioned between 1 N HCI and ethyl acetate. The aqueous layer is basified with 6N sodium hydroxide to pH 12 and extracted with ethyl acetate. The organic phase is dried over sodium sulfate, filtered, and concentrated to give the desired product as a yellow foam. MS: 415 (M + 1). Step E. 2-[[4-[[6-Methyl-4'-(trifluoromethyl)-[1 ,1 '-biphenyl]-2-yl]carbonyl]amino]- phenoxy]ethyl]-carbamic acid, methyl ester

To a solution of the amine from the previous step (180 mg, 0.43 mmol) and triethylamine (88 mg, 0.87 mmol) in DMF at 0 ⁹C is added methyl chloroformate (49 mg, 0.52 mmol). The resulting mixture is stirred at room temperature overnight. The mixture is then partitioned between water and ethyl acetate. The organic solution is washed with brine, dried over sodium sulfate, filtered and concentrated to a residue which is chromatographed on silica gel using a 1 :1 hexane-ethyl acetate mixture as the eluent to give the title product as a white foam, m.p. 48-50 ⁹C.

Example 8

r2-rr5-rrr4^,-(Trifluoromethvn-ri.1'-biphenvn-2-vncarbonvnamino1-2- pyridinyllamino.etrtvπ-carbamic acid, methyl ester

Step A. [2-[(5-Nitro-2-pyridinyl)amino]ethyl]-carbamic acid, t-butyl ester

A mixture of 2-chloro-5-nitropyridine (993.9 mg, 6.27 mmol), sodium acetate (534 mg, 6,51 mmol), triethylamine (653 mg, 6.46 mmol) and N-BOC-ethylenediamine (1000 mg, 6.27 mmol) in 20 mL of ethanol is stirred at 80⁹C for 24 h. The mixture is filtered and concentrated to give a yellow solid residue. The solid is taken up in ethyl acetate and washed with water and brine. The organic solution is dried over magnesium sulfate and concentrated to give desired product as a yellow solid. MS: 283 (M + 1).

Step B. [2-[(5-Amino-2-pyridinyl)amlno]ethyl]- carbamic acid, t-butyl ester

A mixture of the nitropyridine derivative from Step A (1.67 g, 5.91 mmol) and 10% palladium on carbon (200 mg) in 20 mL of THF under 50 psi of hydrogen is shaken for 7 h. The mixture is filtered and concentrated to give desired product as a dark foam.

Step C. [2-[[5-[[[4'-(Trifluoromethyl)-[1 ,1 '-biphenyl]-2-yl]carbonyl]amino]-2-pyridinyl]- amino]ethyl]-carbamic acid, t-butyl ester

A solution of 4'-(trifluoromethyl)-1 ,1'-biphenyl-2-carboxylic acid (1 ,732 mg, 6.51 mmol), EDCI (1 ,282 mg, 6.5 mmol), HOAt (887.2 mg, 6.52 mmol) in 6 mL of DMF is stirred for 1.5 h. Then a solution of the crude aminopyridine derivative from previous Step B (ca. 5.91 mmol) in 8 mL of DMF is added. The reaction is stirred at room temperature for 24 h, then concentrated under vacuum to remove most of the DMF. The residue is taken up in ethyl acetate and washed with water, saturated aqueous sodium bicarbonate, and brine. The combined aqueous extracts are washed once with ethyl acetate, and the combined organic solutions are treated with charcoal, dried with magnesium sulfate and concentrated to dryness. The crude product is chromatographed on silica gel using a hexane-ethyl acetate mixture (1:2) as the eluent to yield a tan solid. Recrvstallization from 1:1 hexane-ethyl acetate gives the desired product as a white solid, m.p. 166-166.5 ²C. Step D. iV-[6-[(2-Aminoethyl)amino]-3-pyridinyl]-4'-(trifluoromethyl)-[1 ,1 '-biphenyl]-2- carboxamide

Trifluoroacetic acid (4.39 g, 38.5 mmol) is added to a suspension of the t-BOC derivative from Step C (1.93 g, 3.85 mmol) in methylene chloride at room temperature. The reaction mixture is stirred 24 h, then concentrated under vacuum. Toluene (4 mL) is added to the residue and the mixture is again concentrated under vacuum. Toluene addition and concentration is repeated twice more, and the residue is taken up in a small amount of water. Sodium carbonate (3 g in 20 mL of water) is added with stirring, and the mixture is extracted with ethyl acetate and MTBE. The organic solution is washed with brine, dried over magnesium sulfate, and concentrated to give desired product as an off-white foam. MS: 401.1 (M + 1).

Step E. [2-[[5-[[[4'-(Trif luoromethyl)-[1 ,1 '-biphenyl]-2-yl]carbonyl]amino]-2- pyridinyl]amino]ethyl]-carbamic acid, methyl ester

CH_jCl,, α00₂CH₃

Methyl chloroformate (39 mg, 0.41 mmol) is added to a mixture of the amine from Step D

133 mg, 0.33 mmol) and polymer-bound morpholine in methylene chloride at room temperature. The reaction mixture is shaken on an orbital shaker for 2.5 h. Then polymer bound tris-(2-aminoethyl)amine (76 mg) and polymer-bound isocyanate (61 mg) and methylene chloride (0.5 mL) are added and shaking is continued for 2.5 h. The mixture is filtered through a fritted glass filter and the solids are washed with THF, acetone, and methylene chloride. The filtrate is concentrated to give a tan oil which is chromatographed on silica gel using a hexane-ethyl acetate mixture (1 :3) as the eluent to yield title product, m.p. 180-182; MS: 459 (M + 1).

Example 9

Prepared similarly to previous examples 5 to 8 are the compounds listed below, using the appropriate starting materials.

The compound of example 9(c) is converted to the compound of example 9(d) as follows:

9 (c) 9 (d)

Sodium hydride (19.5 mg, 0.81 mmol) is added to a solution of the sulfonamide (example 9c, 420 mg, 0.74 mmol) in 15 mL of DMF at 0⁹C. Methyl iodide (105 mg, 0.74 mmol) is then added and the mixture is allowed to warm to room temperature and stirred overnight. The mixture is then partitioned between ethyl acetate and water. The ethyl acetate solution is washed with brine, dried over sodium sulfate, filtered and concentrated to give the crude product. The crude product is purified by flash chromatography on silica gel using 30% ethyl acetate in hexane as the eluent to give the compound of example 9(d). Example lO

Hard gelatin capsules, comprising 50 mg active substance can be prepared for example as follows:

Composition (for 1000 capsules) Active ingredient 50.0 g

Lactose 250.0 g

Microcrystalline cellulose 30.0 g Sodium Lauryl sulfate 2.0 g

Magnesium stearate 8.0 g

The sodium lauryl sulfate is added to the active ingredient. Both components are intimately mixed. Then first the lactose is added and then the microcrystalline cellulose. Thereupon these components are intimately mixed for a further 10 minutes. Finally the magnesium stearate is added. After 3 minutes of further mixing, the formulation is filled into hard gelatin capsules of size 0 (340 mg each).

Claims

What is claimed is:

1. A compound of the formula

O

R.- L— I CI — NH — U— — alk— W — Z (0

wherein

L and L' are arylene or heteroarylene;

Z is aryl or heteroaryl; or when W is NR₀, Z is also

heteroaryl-lower — COOR_d, or — SO₂R_θ in which

R_a, Rd and R₈ are independently optionally substituted lower alkyl, cycloalkyl, adamantyl, aryl, heteroaryl or (aryl or heteroaryl)-lower alkyl; and R_b and R₀ are independently hydrogen, cycloalkyl, optionally substituted lower alkyl, aryl, heteroaryl or (aryl or heteroaryl)-lower alkyl; or R_b and R_c together represent lower alkylene or lower-alkylene interrupted by O, S or N-(H, lower alkyl, acyl or aralkyl);

R₀ is hydrogen, lower alkyl or aryl-lower alkyl; m is zero, 1 or 2; and alk is lower alkylene; or a pharmaceutically acceptable salt thereof.

2. A compound according to claim 1 of the formula

wherein

R is hydrogen, lower alkyl, lower alkoxy, halo or trifluoromethyl; i is aryl, cycloalkyl, heterocyclyl, aryl-lower alkoxy or aryl-lower alkylthio, provided that Ri is not carboxyphenyl when V and W are O or when one of V and W is O and the other is a direct bond, and when Z is aryl or heteroaryl;

L is arylene or heteroarylene;

Y is CH or N;

Z is aryl or heteroaryl; or when W is NR₀, Z is also

heteroaryl-lower alkyl, — COOR_d, or — SO₂R_e in which

R_a, R_d and R_e are independently optionally substituted lower alkyl, cycloalkyl, adamantyl, heteroaryl or (aryl or heteroaryl)-lower alkyl; and R and R₀ are independently hydrogen, cycloalkyl, optionally substituted lower alkyl, aryl, heteroaryl or (aryl or heteroaryl)-lower alkyl; or R_b and R_c together represent lower alkylene or lower-alkylene interrupted by O, S or N-(H, lower alkyl, acyl or aralkyl);

R₀ is hydrogen, lower alkyl or aryl-lower alkyl; m is zero, 1 or 2; and n is an integer from 1 -4 provided that either V or W is a direct bond when n is 1 ; or a pharmaceutically accepfable sairthereof:

3. A compound according to claim 1 of the formula

wherein X is N or CR₂;

Y is N or CH;

R is hydrogen, lower alkyl, lower alkoxy, halo or trifluoromethyl;

R₀ is hydrogen or lower alkyl;

R-i is aryl, cycloalkyl, heteroaryl, aryl-lower alkoxy or aryl-lower alkylthio; provided that Ri is not carboxyphenyl when both V and W are O or when one of V and W is O and the other is a direct bond;

Ar is monocyclic aryl or heteroaryl;

R₂, R₃, R₄, and R₅ are independently hydrogen, optionally substituted alkyl, halo, amino, substituted amino, trifluoromethyl, cyano, carboxyl, alkoxycarbonyl, aralkoxycarbonyl, (alkyl, aryl or aralkyl)-thio, (alkyl, aryl, or aralkyl)-oxy, acyloxy, (alkyl, aryl or aralkyl)-aminocarbonyloxy; or any two or R₂, R₃, R₄ and R₅ at adjacent positions are alkylenedioxy; n is 2 or 3; m is zero; or a pharmaceutically acceptable salt thereof.

4. A compound according to claim 3 of formula III wherein V is O, S(O)_m or NR₀, and W is a direct bond; or a pharmaceutically acceptable salt thereof.

5. A compound according to claim 3 of formula III wherein R is phenyl or phenyl-lower alkoxy, each optionally substituted on phenyl by lower alkyl, lower alkoxy, halo, trifluoromethyl or cyano; X is CR₂; R, R₂, R₃, R₄ and R₅ are independently hydrogen, lower alkyl, lower alkoxy, halo or trifluoromethyl; Y is CH or N; V is NH, N-CH₃ or O; n is 1 , 2 or 3; W is a direct bond; and Ar is phenyl or phenyl substituted by lower alkyl, halo, trifluoromethyl, lower alkoxy or cyano; or Ar is thienyl, pyridyl, indolyl or pyrimidyl optionally substituted by lower alkyl; or a pharmaceutically acceptable salt thereof.

6. A compound according to claim 3 of formula 111 wherein R is hydrogen, methyl or chloro; Ri is trifluoromethylphenyl or benzyloxy; Y is CH or N; X is CR₂; R₂ is methyl; R₃, R₄ and R₅ are hydrogen; V is NH; n is 2 or 3; W is a direct bond; Ar is phenyl or 2-pyridyl; or a pharmaceutically acceptable salt thereof.

7. A compound according to claim 5 of formula III wherein R is hydrogen; Ri is 4- trifluoromethylphenyl; Y is CH; X is CR₂; R₂ is methyl; R₃-R₅ are hydrogen; V is O; n is 2; W is a direct bond; and Ar is 2-pyridyl; or a pharmaceutically acceptable salt thereof.

8. A compound according to claim 2

wherein X is N or CR₂;

Y is N orCH;

V is O, S(O)_m, NR_o or a direct bond;

R is hydrogen, lower alkyl, lower alkoxy, halo or trifluoromethyl;

R₀ is hydrogen or lower alkyl;

Ri is aryl. cycloalkyl, heteroaryl, aryl-lower alkoxy or aryl-lower alkylthio;

R₂, R₃, R₄, and R₅ are independently hydrogen, lower alkyl, halo, trifluoromethyl, cyano, (lower alkyl, aryl, or aralky!)-thio, (lower alkyl, aryl, or aralkyl)-oxy, acyloxy, (alkyl, aryl or aralkyl)-aminocarbonyloxy; or any two or R₂, R₃, R₄ and R5 at adjacent positions are alkylenedioxy; Z is heteroaryl-lower alkyl, — COR_a,-CON^ , — COOR_d, or — SO₂R_e in which _c

R_a, R_d and R_e are independently optionally substituted lower alkyl, cycloalkyl, adamantyl, aryl, heteroaryl or (aryl or heteroaryl)-lower alkyl; and R_b and R_c are independently hydrogen, cycloalkyl, optionally substituted lower alkyl, aryl, heteroaryl or (aryl or heteroaryl)-lower alkyl; or R_b and R₀ together represent lower alkylene or lower-alkylene interrupted by O, S or N-(H, lower alkyl, acyl or aralkyl); m is zero, 1 or 2; and n is an integer from 1-4; or a pharmaceutically acceptable salt thereof.

9. A compound according to claim 8 of formula IV wherein Ri is phenyl or phenyl-lower alkoxy, each optionally substituted on phenyl by lower alkyl, lower alkoxy, halo, trifluoromethyl or cyano; X is CR₂; Y is CH or N; R, R₂, R₃, R and R₅ are independently hydrogen, lower alkyl, lower alkoxy, halo or trifluoromethyl; R₀ is hydrogen; V is a direct bond, NH, N-CH₃ or O; n is 2 or 3; Z is _b heteroaryl-lower alkyl,— CON , ^— COOR_d or — SO₂R_e; _c

R_b and R_c are lower alkyl; or R_b and R₀ together represent lower alkylene or lower alkylene interrupted by O, S, N-(H, lower alkyl or aralkyl);

R_d is lower alkyl or aryl -lower alkyl;

R_e is lower alkyl, monocyclic aryl or monocyclic aryl-lower alkyl;or a pharmaceutically acceptable salt thereof.

10. A compound according to claim 9 of formula IV wherein R is hydrogen or chloro; Ri is Jrifluor-oιifietrxylrjheιyl;_X s CR?; Y is CH or N: is methyl: R_a. R-t and R₅ are hydrogen: Ro is hydrogen; V is NH, O, or a direct bond; n is 2 or 3; and Z is -COOR_d wherein R_d is lower alkyl; or a pharmaceutically acceptable salt thereof.

11. A pharmaceutical composition comprising a compound according to any one of claims 1-10 or a pharmaceutically acceptable salt thereof and one or more pharmaceutically acceptable carriers.

12. Use of a compound according to any one of claims 1-10 or of a pharmaceutically acceptable salt thereof for the preparation of a medicament for treating microsomal triglyceride transfer protein or lipoprotein B dependent conditions in mammals.

13. Use according to claim 12 for the preparation of a medicament for treating atherosclerosis, hypertriglyceridemia or hypercholesteremia.

14. A method of treating microsomal triglyceride transfer protein or lipoprotein B dependent conditions in mammals which comprises administering to a mammal in need thereof an effective amount of a compound according to any one of claims 1 -10 or a pharmaceutically acceptable salt thereof.