HK1083451B - N-aryl piperidine substituted biphenylcarboxamides as inhibitors of apolipoprotein b secretion - Google Patents

Description

N-aryl piperidine substituted biphenylcarboxamides as inhibitors of apolipoprotein B secretion

The present invention relates to novel N-aryl piperidine substituted biphenylcarboxamides having apolipoprotein B inhibitory activity and concomitant lipid lowering activity. The invention also relates to processes for the preparation of such compounds, pharmaceutical compositions comprising said compounds and the use of the compounds as a medicine for the treatment of hyperlipidemia, obesity and type II diabetes.

Obesity is the cause of a myriad of serious health problems, such as adult onset of diabetes and heart disease. In addition, weight loss is becoming an important focus for more people.

The causal relationship between hypercholesterolemia, particularly that associated with increased plasma concentrations of low density lipoproteins (hereinafter abbreviated as LDL) and very low density lipoproteins (hereinafter abbreviated as VLDL), and early atherosclerosis and/or cardiovascular disease has now been widely observed. However, there are currently only a limited number of drugs for treating hyperlipidemia.

Drugs mainly used for the control of hyperlipidemia include bile acid sequestrant resins such as cholestyramine and lipidol No. II, fibric acid derivatives such as bezafibrate, clofibrate, fenofibrate, ciprofibrate and gemfibrozil, nicotinic acid and cholesterol synthesis inhibitors such as HMG coenzyme-a reductase inhibitors. There remains a need, however, for new lipid lowering agents that have improved efficacy and/or action over the above-mentioned drugs via other mechanisms.

Plasma lipoproteins are high molecular weight water-soluble complexes of fats (cholesterol, triglycerides, phospholipids) and apolipoproteins. The five major classes of lipoproteins, which differ in the proportion of fat and the type of apolipoprotein, all originate from the liver and/or the intestine and have been defined in terms of their density (measured by ultracentrifugation). It includes LDL, VLDL, intermediate density lipoprotein (hereinafter abbreviated as IDL), high density lipoprotein (hereinafter abbreviated as HDL) and chylomicron. Ten major human plasma apolipoproteins have been identified. VLDL, which is secreted from the liver and contains apolipoprotein B (hereinafter abbreviated as Apo-B), undergoes a degradation reaction to give LDL, which can transport 60 to 70% of the total amount of serum cholesterol. Apo-B is also the major protein component of LDL. Increased LDL-cholesterol in plasma is a cause of atherosclerosis due to excessive synthesis or decreased metabolism. In contrast, high density lipoprotein (hereinafter abbreviated HDL) having apolipoprotein Al has a protective effect and is conversely associated with the prevalence of coronary heart disease. Thus, the HDL/LDL ratio is a convenient method for assessing the likelihood that an individual's plasma lipid profile will cause atheroma.

Two isomeric forms of apolipoprotein (apo) B: apo B-48 and apo B-100 are important proteins in human lipoprotein metabolism. Apo B-48, named Apo B-48, was synthesized by the human intestine as approximately 48% of the Apo B-100 size on sodium dodecyl sulfate-polyacrylamide gels. Apo B-48 is essential for the composition of chylomicrons and therefore plays an essential role in the intestinal absorption of dietary fat. ApoB-100 produced by the human liver is required for the synthesis and secretion of VLDL. LDL, which contains approximately 2/3 cholesterol in human plasma, is a metabolite of VLDL. Apo B-100 is also practically the only protein component of LDL. Elevated plasma concentrations of apo B-100 and LDL cholesterol were identified as risk factors for developing atherosclerotic coronary artery disease.

Hyperlipidemia can be caused by a wide variety of inherited and acquired diseases. It can be classified into primary and secondary hyperlipidemic states. The most common causes of secondary hyperlipidemia are diabetes, alcohol abuse, medications, hypothyroidism, chronic renal failure, nephrotic syndrome, cholestasis, and bulimia. Primary hyperlipidemia can also be classified into general hypercholesterolemia, familial combined hyperlipidemia, familial hypercholesterolemia, residual hyperlipidemia, chylomicronemia syndrome, and familial hypertriglyceridemia.

Microsomal triglyceride transfer protein (hereinafter abbreviated as MTP) is known to facilitate transport of triglycerides and cholesterol esters by selection of phospholipids, such as phosphatidylcholine. Sharp, et al Nature (1993) 365: 65 the defect responsible for the beta-lipoprotein deficiency was demonstrated to be located in the MTP gene. This indicates that MTP is essential for the synthesis of Apo B-containing lipoproteins (e.g. VLDL, a precursor of LDL). Thus, the MTP inhibitor will then inhibit the synthesis of VLDL and LDL, thereby lowering the concentration of VLDL, LDL, cholesterol and triglycerides in humans.

It is an object of the present invention to provide an improved treatment for patients suffering from obesity or atherosclerosis, particularly coronary atherosclerosis and more generally diseases associated with atherosclerosis, such as ischemic heart disease, peripheral vascular disease and cerebrovascular disease. It is another object of the invention to produce regression of atherosclerosis and inhibit its clinical consequences, particularly morbidity and mortality.

MTP inhibitors have been disclosed in WO 00/32582, WO 01/96327 and WO 02/20501.

The present invention is based on the unexpected finding that: a novel class of N-aryl piperidine substituted biphenylcarboxamide compounds are useful as selective MTP inhibitors, i.e., capable of selectively blocking MTP at the intestinal wall level in mammals, and are therefore promising as pharmaceutical agents, i.e., for the treatment of hyperlipidemia. The present invention further provides several methods of making the N-aryl piperidine substituted biphenylcarboxamide compounds, as well as pharmaceutical compositions containing such compounds. Also, the present invention provides some novel compounds which are useful intermediates for preparing therapeutically active N-aryl piperidine substituted biphenylcarboxamide compounds, and methods for preparing the intermediates. Finally, the present invention provides a method of treating a condition selected from the group consisting of atherosclerosis, pancreatitis, obesity, hypercholesterolemia, hypertriglyceridemia, hyperlipidemia, diabetes and type II diabetes comprising administering a therapeutically active biphenylcarboxamide compound to a mammal.

The invention relates to a novel compound of formula (I)

N-oxides, pharmaceutically acceptable acid addition salts and stereochemically isomeric forms thereof, wherein

R¹Is hydrogen, C_1-4Alkyl, halogen, or polyhaloC_1-4An alkyl group;

R²is hydrogen, C_1-4Alkyl, halogen, or polyhaloC_1-4An alkyl group;

R³is hydrogen or C_1-4An alkyl group;

R⁴is hydrogen, C_1-4Alkyl or halogen;

n is an integer of 0 or 1;

X¹and X²Or both are carbon, or when X¹And X²When one is nitrogen, X¹And X²The other of (a) and (b) is carbon;

X³is carbon or nitrogen, provided that X¹And X²Only one of which is nitrogen;

y is O or NR⁶Wherein R is⁶Is hydrogen or C^1-4An alkyl group; and

R⁵is hydrogen, optionally substituted by C_1-4Alkoxy, cyano, polyhaloC_1-4Alkyl or aryl substituted C_1-6An alkyl group; c optionally substituted by aryl_2-6An alkenyl group; c optionally substituted by aryl_3-6An alkynyl group;

aryl or heteroaryl;

aryl is phenyl; is one, two or three independently selected from nitro, azido, cyano, halogen, hydroxyl, C_1-6Alkyl radical, C_3-6Cycloalkyl radical, C_1-4Alkoxy, polyhalo C_1-6Alkyl, amino, mono-or di (C)_1-6Alkyl) phenyl substituted with a substituent of the amino group;

heteroaryl is pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, triazinyl, triazolyl, imidazolyl, pyrazolyl, thiazolyl, isothiazolyl, oxazolyl, pyrrolyl, furanyl or thienyl; and optionally substituted by one, two or three independently selected from nitro, azido, cyano, halogen, hydroxy, C_1-6Alkyl radical, C_3-6Cycloalkyl radical, C_1-4Alkoxy, polyhalo C_1-4Alkyl, amino, mono-or di (C)_1-6Alkyl) amino;

unless otherwise indicated, the above definitions and below:

halo generally refers to fluoro, chloro, bromo and iodo;

-C_1-4alkyl means a straight or branched chain saturated hydrocarbon group having 1 to 4 carbon atoms, such as methyl, ethyl, propyl, n-butyl, 1-methylethyl, 2-methylpropyl, 1-dimethylethyl and the like;

-C_1-6the alkyl group comprising C_1-4Alkyl (as previously defined) and its higher homologues having 5 or 6 carbon atoms, such as 2-methylbutyl, n-pentyl, dimethylpropyl, n-hexyl, 2-methylpentyl, 3-methylpentyl and the like;

-polyhalo C_1-4Alkyl is defined as polyhalo-substituted C_1-4Alkyl, especially C, substituted by 2 to 6 halogen atoms_1-4Alkyl (as previously defined) such as difluoromethyl, trifluoromethyl, trifluoroethyl, and the like;

-C_2-6alkenyl is defined as a straight-chain or branched unsaturated hydrocarbon radical having 2 to 6 carbon atoms, e.g. ethenyl, propenyl, butenyl, pentenA phenyl or hexenyl group;

-C_3-6alkynyl is defined as a straight or branched chain hydrocarbon group having 3 to 6 carbon atoms and having a triple bond, such as 2-propynyl, 3-butynyl, 2-pentynyl, 3-methyl-2-butynyl, 3-hexynyl, 2-hexynyl, and the like;

-C_1-4alkylamino is defined as a primary amino group having 1 to 6 carbon atoms, such as methylamino, ethylamino, propylamino, isopropylamino, butylamino, isobutylamino, and the like;

-bis (C)_1-6Alkyl) amino is defined as a secondary amino group having 1 to 6 carbon atoms, such as dimethylamino, diethylamino, dipropylamino, diisopropylamino, N-methyl-N '-ethyl-amino, N-ethyl-N' -propyl-amino, and the like.

The pharmaceutically acceptable acid addition salts hereinbefore defined include the therapeutically active, non-toxic acid addition salt forms which the compounds of formula (I) are able to form. The pharmaceutically acceptable acid addition salts can be readily obtained by treating the base with the appropriate acid. Suitable acids include, for example, inorganic acids, e.g., hydrohalic acids, such as hydrochloric or hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and the like; or organic acids such as acetic acid, propionic acid, glycolic acid, lactic acid, pyruvic acid, oxalic acid (i.e., oxalic acid), malonic acid, succinic acid (i.e., succinic acid), cis-butenedioic acid, trans-butenedioic acid, malic acid, tartaric acid, citric acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, cyclamic acid, salicylic acid, p-aminosalicylic acid, pamoic acid (pamoic acid), and the like.

Conversely, the salt form may be converted to the free base form by treatment with a suitable base.

The term addition salt as used hereinbefore also includes the solvates which the compounds of formula (I) and salts thereof are able to form. Such solvates are for example hydrates, alcoholates and the like.

The N-oxide forms of the compounds of formula (I) may be prepared by methods known in the art, including compounds of formula (I) in which one nitrogen atom is oxidized to the N-oxide.

The term "stereochemically isomeric forms" as used hereinbefore refers to all possible stereoisomeric forms which the compounds of formula (I) may contain. Unless otherwise mentioned or indicated, the chemical designation of a compound denotes the mixture of all possible stereochemically isomeric forms, which mixtures contain all diastereomers and enantiomers of the basic molecular structure. More particularly, the stereocenter may have the R-or S-configuration; the substituents on the divalent cyclic (partially) saturated groups may have either the cis-or trans-configuration. Unless otherwise indicated, the chemical designation of a compound denotes the mixture of all possible stereochemically isomeric forms, which mixtures contain all diastereomers and enantiomers of the basic molecular structure. The same applies to the intermediates disclosed herein for the preparation of the final product of formula (I).

Cis and trans are used herein consistent with chemical abstracts nomenclature and refer to the position of a substituent on the ring moiety.

The absolute stereochemical configuration of the compounds of formula (I) and intermediates used in their preparation can be readily determined by those skilled in the art using well known methods, such as X-ray diffraction.

Also, some compounds of formula (I) and some intermediates used in the preparation of formula (I) may exhibit polymorphism. It is to be understood that the present invention encompasses any polymorph form useful in the treatment of the conditions noted above.

An attractive group of compounds includes compounds of formula (I) wherein one or more of the following limitations apply:

a)R¹is tert-butyl or trifluoromethyl;

b)R²is hydrogen or C_1-4An alkyl group;

c)R³is hydrogen;

d)R⁴is hydrogen;

e)R⁵is C_1-4Alkyl or phenyl substituted C_1-4An alkyl group;

a first group of specific compounds is X in formula (I)¹、X²And X³A compound that is carbon.

A second specific group of compounds is X in formula (I)¹Is carbon, X²Is nitrogen and X³A compound that is carbon.

A third specific group of compounds is X in formula (I)¹Is nitrogen, X²Is carbon and X³A compound that is carbon.

A fourth specific group of compounds of formula (I) wherein X¹Is carbon, X²Is nitrogen and X³A compound that is nitrogen.

A fifth specific group of compounds are those of formula (I) wherein n is an integer 0.

A sixth specific group of compounds are those of formula (I) wherein n is an integer 1.

A first group of preferred compounds is that of formula (I) R¹Is C_1-4Alkyl or trifluoromethyl; r²Is hydrogen or C_1-4An alkyl group; r³Is hydrogen; r⁴Is hydrogen; r⁵Is C_1-4Alkyl or phenyl substituted C_1-4An alkyl group; n is an integer of 0; and X¹、X²And X³A compound that is carbon.

A second group of preferred compounds is that of formula (I) R¹Is C_1-4Alkyl or trifluoromethyl; r²Is hydrogen or C_1-4An alkyl group; r³Is hydrogen; r⁴Is hydrogen; r⁵Is C_1-4Alkyl or phenyl substituted C_1-4An alkyl group; n is an integer of 1; and X¹、X²And X³A compound that is carbon.

A third group of preferred compounds is that of formula (I) R¹Is C_1-4Alkyl or triA fluoromethyl group; r²Is hydrogen or C_1-4An alkyl group; r³Is hydrogen; r⁴Is hydrogen; r⁵Is C_1-4Alkyl or phenyl substituted C_1-4An alkyl group; n is an integer of 0; x³Is carbon and X¹Or X²Is nitrogen, and X¹Or X²Is a carbon compound.

A fourth preferred group of compounds is R in formula (I)¹Is C_1-4Alkyl or trifluoromethyl; r²Is hydrogen or C_1-4An alkyl group; r³Is hydrogen; r⁴Is hydrogen; r⁵Is C_1-4Alkyl or phenyl substituted C_1-4An alkyl group; n is an integer of 1; x³Is carbon and X¹Or X²Is nitrogen, and X¹Or X²Is a carbon compound.

The first group of more preferred compounds is the group of preferred compounds wherein Y is O.

A second group of more preferred compounds is the group of preferred compounds wherein Y is NH.

A first process for the preparation of the compounds of formula (I) is, wherein an intermediate of formula (II)

Wherein R is³、R⁴、R⁵、n、Y、X¹、X²And X³Biphenyl carboxylic acids or halides of the formula (III), as defined for the formula (I)

Wherein R is¹、R²Is as defined in formula (I) and Q¹Selected from hydroxyl and halogen, in at least one reaction-inert solventIn the presence of a suitable base, said process additionally optionally comprising the conversion of a compound of formula (I) into an addition salt thereof, and/or the preparation of a stereochemically isomeric form thereof. When Q is¹In the case of hydroxyl, the biphenyl carboxylic acid of formula (III) can be conveniently activated by adding an effective amount of a reaction promoter. Non-limiting examples of such reaction promoters include: carbonyldiimidazole, diimides, such as N, N '-Dicyclohexylcarbodiimide (DCC) or 1-ethyl-3- (3' -dimethylaminopropyl) -carbodiimide (ECC), and functional derivatives thereof. For this acylation process, a polar aprotic solvent, such as dichloromethane, is preferably used. Suitable bases for carrying out this first process include tertiary amines such as triethylamine, triisopropylamine and the like. Depending on the particular solvent used, suitable temperatures in the first process are generally from about 20 ℃ to about 140 ℃, and most typically the melting point of the solvent.

The second process for preparing the biphenylcarboxamide compounds of the invention is one in which an intermediate of the formula (IV)

Wherein R is¹、R²、R³、n、Y、X¹、X²And X³Is as defined for formula (I), and Q²Selected from halogen and hydroxy, and of the formula R⁵Intermediates (V) of-Y-H, wherein R⁵The reaction is carried out in at least one reaction-inert solvent and optionally in the presence of at least one suitable coupling agent and/or a suitable base, as defined for Y in formula (I), said process additionally optionally comprising converting the compound of formula (I) into an addition salt thereof, and/or preparing a stereochemically isomeric form thereof. When Q is²In the case of hydroxyl groups, the carboxylic acid of formula (IV) may conveniently be activated by the addition of an effective amount of a reaction promoter. Non-limiting examples of such reaction promoters include: carbonyldiimidazoles, imides, e.g. DCC, ECC, hydroxybenzotriazole, benzotriazol-1-yl-N-oxytris- (dimethylamino) *Hexafluorophosphate (BOP), tetrapyrrolidine * hexafluorophosphate, bromotripyrrolidine * hexafluorophosphate, or functional derivatives thereof, for example, as disclosed in "Solid-Phase Synthesis: a practical guide, edited by Steven A. Kates and Fernando Albericio, Marcel Dekker, 2000 (ISBN: 0-8247-.

A third process for preparing the biphenylcarboxamide compounds of the invention is one in which an intermediate of the formula (VI)

Wherein R is¹、R²、R³、R⁴、X¹、X²And X³Is as defined for formula (I), and Q³Selected from halogen, B (OH)₂Alkyl borates (alkylboronates) and cyclic analogues thereof, with reactants of formula (VII)

Wherein n, Y and R⁵The reaction is carried out in at least one reaction-inert solvent and optionally in the presence of at least one transition metal coupling agent and/or at least one suitable ligand, as defined for formula (I), said process additionally optionally comprising converting the compound of formula (I) into an addition salt thereof, and/or preparing a stereochemically isomeric form thereof. Such reactions are known in the art as Buchwald's reactions, and for metal coupling agents and/or suitable ligands that may be used, e.g., palladium compounds, e.g., palladium tetrakis (triphenylphosphine), tris (dibenzylidene-acetonedipalladium, 2 ' -bis (diphenylphosphino) -1, 1 ' -Binaphthyl (BINAP), etc., see, e.g., Tetrahedron Letters, (1996), 37(40), 7181-³Is B (OH)₂According to Tetrahedron Letters, (1998)),39: 2933-6, an alkyl borate ester or its cyclic analogue and then copper acetate as a coupling agent can be used.

A compound of formula (I-a) defined wherein Y represents NH and R³Compounds of formula (I) which represent hydrogen may conveniently be prepared using solid phase synthesis methods as shown in scheme 1 below. Typically, solid phase synthesis involves reacting intermediates in the synthesis with a polymeric support. These polymer-supported intermediates can then be subjected to multiple synthetic steps. After each step, impurities are removed by filtering the resin and washing multiple times with various solvents. In each step, the resin can be isolated and reacted with various intermediates in the next step, whereby a variety of compounds can be synthesized. After the final step of the method, the resin is treated with a reagent or separated from the sample by processing. Further details of methods of use in solid Chemistry are disclosed, for example, in "Handbook of Combinatorial Chemistry: drugs, catalysis, Materials ", K.C.Nicolaou, R.Hanko, and W.Hartwig, volumes 1 and 2, Wiley (ISBN: 3-527-.

Scheme 1:

the abbreviations used in scheme 1 are explained in the experimental section. Substituent R¹、R²、R³、R⁴、R⁴、R⁵、n、Y、X¹、X²And X³The same as defined in formula (I). PG represents a protecting group, e.g. C_1-6Alkoxycarbonyl, phenylmethoxycarbonyl, t-butoxycarbonyl, 9-fluorenylmethoxycarbonyl (Fmoc), and the like.

A compound of formula (I-b), defined as wherein R³The compounds of formula (I), which represent hydrogen, can be prepared using solid phase synthesis methods as shown in scheme 2 below.

Scheme 2

The compounds of formula (I) prepared in the processes disclosed hereinabove may be synthesized as racemic mixtures of enantiomers, which can be separated from one another by known resolution methods. Racemic compounds of formula (I) can be converted into their corresponding diastereomeric salt forms by reaction with an appropriate chiral acid. The diastereomeric salt forms are then separated, for example, by selective or fractional crystallization and the enantiomers are separated therefrom with a base. Another method for separating the enantiomers of the compound of formula (I) includes liquid chromatography using a chiral stationary phase. If this reaction occurs stereospecifically, the pure stereochemically isomeric forms may also be prepared from the corresponding pure stereochemically isomeric forms of the appropriate starting materials. If a particular stereoisomer is desired, the compound will preferably be synthesized by stereospecific methods of preparation. These processes will advantageously use enantiomerically pure starting materials.

N-aryl piperidine substituted biphenylcarboxamides of formula (I), their N-oxide forms, pharmaceutically acceptable salts and stereoisomeric forms thereof have advantageous apolipoprotein B inhibitory activity and concomitant lipid lowering activity. The compounds of the invention are therefore useful in medicine, in particular in a method for treating a patient suffering from hyperlipidemia, obesity, atherosclerosis or type II diabetes. The compounds of the invention are particularly useful in the manufacture of medicaments for the treatment of diseases caused by an excess of Very Low Density Lipoprotein (VLDL) or Low Density Lipoprotein (LDL), especially diseases caused by cholesterol associated with said VLDL and LDL.

The main mechanism of action of the compounds of formula (I) appears to include inhibition of MTP (microsomal triglyceride transfer protein) activity in hepatocytes and intestinal epithelial cells, resulting in a reduction in VLDL and chylomicron production, respectively. This is a new and innovative approach for hyperlipidemia and is expected to reduce LDL-cholesterol and triglycerides by reducing VLDL production in the liver and chylomicrons in the intestine.

Hyperlipidemia can be caused by a wide variety of inherited and acquired diseases. It can be classified into primary and secondary hyperlipidemic states. The most common causes of secondary hyperlipidemia are diabetes, alcohol abuse, medications, hypothyroidism, chronic renal failure, nephrotic syndrome, cholestasis, and bulimia. The primary hyperlipidemia is hypercholesterolemia in general, familial combined hyperlipidemia, familial hypercholesterolemia, residual hyperlipidemia, chylomicronemia syndrome and familial hypertriglyceridemia. The compounds of the invention can be used for the prevention or treatment of patients suffering from obesity or atherosclerosis, in particular coronary atherosclerosis and more generally diseases associated with atherosclerosis, such as ischemic heart disease, peripheral vascular disease, cerebrovascular disease. The compounds of the invention can cause regression of atherosclerosis and inhibit the clinical consequences of atherosclerosis, especially morbidity and mortality.

In view of the effects of the compounds of formula (I), the present invention also provides a method of treating warm-blooded animals, including humans (generally referred to herein as patients), suffering from a disease caused by excess Very Low Density Lipoprotein (VLDL) or Low Density Lipoprotein (LDL), particularly a disease caused by cholesterol associated with said VLDL and LDL. Accordingly, the present invention provides a method of treatment to alleviate a condition in a patient suffering from, for example, hyperlipidemia, obesity, atherosclerosis, or type II diabetes.

Apo B-48 synthesized by the intestine is essential for the composition of chylomicrons and plays an essential role in the intestinal absorption of dietary fat. The present invention provides biphenyl carboxamide compounds as selective MTP inhibitors at the intestinal wall stage.

In addition, the present invention provides pharmaceutical compositions comprising at least one pharmaceutically acceptable carrier and a therapeutically effective amount of an N-aryl piperidine substituted biphenylcarboxamide compound of formula (I).

To prepare the pharmaceutical compositions of this invention, an effective amount of the particular compound, in base or addition salt form, as the active ingredient is combined in intimate admixture with at least one pharmaceutically acceptable carrier, which carrier may take a wide variety of forms depending on the form of preparation desired for administration. Such pharmaceutical compositions are desirably in unit dosage form preferably suitable for oral, rectal, transdermal or parenteral administration.

For example, when preparing compositions for oral administration, any of the usual liquid pharmaceutical carriers may be employed, e.g., water, glycols, oils, alcohols, and the like in the case of oral liquid preparations such as suspensions, syrups, elixirs, and solutions; or in the case of powders, pills, capsules and tablets, solid pharmaceutical carriers such as starches, sugars, kaolin, lubricants, binders, disintegrating agents and the like may be employed. Because of their ease of administration, tablets and capsules represent the most advantageous oral dosage unit form in which case solid pharmaceutical carriers are obviously employed. For parenteral injection compositions, the pharmaceutical carrier will consist essentially of sterile water, although other ingredients may be included to enhance the solubility of the active ingredient. Injectable solutions may be prepared, for example, using a pharmaceutical carrier comprising saline solution, dextrose solution, or a mixture of both. Injectable suspensions may also be prepared using suitable liquid carriers, suspending agents and the like. In compositions suitable for transdermal administration, the pharmaceutical carrier may optionally include a penetration enhancer and/or a suitable wetting agent, optionally mixed with suitable additives present in small proportions that do not cause significant damage to the skin. The additives may be selected to facilitate administration of the active ingredient to the skin and/or to aid in the preparation of the desired composition. Such topical compositions may be administered in a variety of ways, for example in the form of a transdermal patch, spot-on or ointment. Since the addition salts of the compounds of formula (I) have an increased water solubility compared to their corresponding base forms, they are obviously more suitable for the preparation of aqueous compositions.

It is particularly advantageous to formulate the pharmaceutical compositions of the present invention in unit dosage form for ease of administration and uniformity of dosage. As used herein, "dosage unit form" refers to physically discrete units suitable as unitary dosages, each unit containing a predetermined quantity of active ingredient calculated to produce the desired therapeutic effect in association with the required pharmaceutical carrier. Examples of such unit dosage forms are tablets (including scored or coated tablets), capsules, pills, powder packets, film tablets, solutions or suspensions for injection, teaspoonfuls, tablespoonfuls and the like, as well as isolated polyploids thereof.

For oral administration, the pharmaceutical compositions of the present invention may be in solid dosage forms, e.g., tablets (swallowable and chewable forms), capsules or gelcaps, prepared using conventional methods with pharmaceutically acceptable excipients and carriers, e.g., binders (e.g., pregelatinized corn starch, polyvinylpyrrolidone, hydroxypropylmethylcellulose, and the like), fillers (e.g., lactose, microcrystalline cellulose, calcium phosphate, and the like), lubricants (e.g., magnesium stearate, talc, silica, and the like), disintegrants (e.g., potato starch, sodium glycolate starch, and the like), wetting agents (e.g., sodium lauryl sulfate, and the like), and the like. Such tablets may be coated using methods well known in the art.

Liquid preparations for oral administration may be in the form of, for example, solutions, syrups or suspensions, or they may be presented as a dry product for admixture with water and/or other suitable solution carriers before use. Such liquid preparations may be prepared by conventional methods, optionally together with other pharmaceutically acceptable additives such as suspending agents (e.g. sorbitol syrup, methyl cellulose, hydroxypropylmethyl cellulose or hydrogenated edible fats), emulsifying agents (e.g. lecithin or acacia), non-aqueous carriers (e.g. almond oil, oily esters or ethanol), sweetening agents, flavouring agents, masking agents and preservatives (e.g. methyl or propyl p-hydroxybenzoate or sorbic acid).

The pharmaceutically acceptable sweeteners which may be used in the pharmaceutical composition of the present invention preferably comprise at least one intense sweetener such as aspartame, acesulfame potassium, sodium cyclamate, alitame, dihydrochalcone sweeteners, monellin, stevioside sucralose (4, 1 ', 6' -trichloro-4, 1 ', 6' -trideoxygalactosucrose) or, preferably, saccharin, sodium saccharin or calcium, optionally with the addition of at least one bulk sweetener such as sorbitol, mannitol, fructose, sucrose, maltose, isomalt (isomalt), glucose, hydrogenated glucose syrup, xylitol, caramel or honey. Intense sweeteners can be readily used at low concentrations. For example, sodium saccharin, and the concentration may be from about 0.04% to about 0.1% (weight/volume) of the final formulation. Bulk sweeteners may be used in a wide concentration range of about 10% to about 35%, preferably about 10% to 15% (weight/volume).

A pharmaceutically acceptable flavour which can mask bitter components present in the formulation at low doses, preferably a fruit flavour, such as cherry, raspberry, blackcurrant or strawberry flavour. The combination of two fragrances produced very good results. In high dose formulations, stronger pharmaceutically acceptable flavours are required, for example caramel chocolate (CaramelChocolate), Mint Cool (Mint Cool), Fantasy and the like. Each of the flavoring agents may be present in the final composition at a concentration ranging from about 0.05% to 1% (weight/volume). Combinations of the strong fragrances may be advantageously used. It is preferred to use flavorants which do not cause a change or loss in taste and/or color in the formulation.

The N-aryl piperidine substituted biphenylcarboxamide compounds of the invention may be formulated for parenteral administration by injection, conveniently intravenously, intramuscularly or subcutaneously, for example by bolus injection (bolus injection) or continuous intravenous injection. Formulations for injection may be presented in unit dosage form, e.g., in ampoules or in multi-dose containers, with an added preservative. It may be in the form of a suspension, solution or emulsion in an oily or aqueous carrier and may contain formulatory agents such as isotonic agents, suspending agents, stabilizing agents and/or dispersing agents. Alternatively, the active ingredient may be presented in powder form for constitution with a suitable vehicle, e.g., sterile pyrogen-free water, before use. The biphenylcarboxamide compounds of the present invention may also be formulated in rectal compositions such as suppositories or retention enemas, e.g., containing conventional suppository bases such as cocoa butter and/or other glycerides.

The N-aryl piperidine substituted biphenylcarboxamide compounds of the present invention may be used in combination with other pharmaceutical preparations, in particular the pharmaceutical compositions of the present invention may additionally comprise at least one other lipid lowering agent, thereby achieving a so-called combined lipid lowering therapy. The additional lipid lowering agent may be, for example, a conventionally used known drug for the control of hyperlipidemia, such as bile acid sequestrant resins, fibric acid derivatives or nicotinic acid as described in the background of the invention above. Suitable additional lipid lowering agents may also include other cholesterol biosynthesis inhibitors and cholesterol absorption inhibitors, especially HMG-CoA reductase inhibitors and HMG-CoA synthetase inhibitors, HMG-CoA reductase gene expression inhibitors, CETP inhibitors, ACAT inhibitors, squalene synthetase inhibitors and the like.

Any HMG-CoA reductase inhibitor may be used as the second compound in the combination therapy aspect of the invention. The term "HMG-CoA reductase inhibitor" as used herein, unless otherwise indicated, refers to a compound that inhibits the biotransformation of hydroxymethylglutaryl-coenzyme a to mevalonate catalyzed by the enzyme HMG-CoA reductase. One skilled in the art can readily determine this inhibition according to standard analytical Methods, namely Methods of enzymology (1981) 71: 455-. Typical compounds are disclosed in, for example, US4,231,938 (including lovastatin), US4,444,784 (including simvastatin), US4,739,073 (including fluvastatin), US4,346,227 (including pravastatin), EP- ｃA-491,226 (including rivastatin) and US4,647,576 (including atorvastatin).

Any HMG-CoA synthetase inhibitor may be used as the second compound in the combination therapy aspect of the invention. The term "HMG-CoA synthase inhibitor" as used herein, unless otherwise specified, refers to a compound that inhibits the biosynthesis of hydroxymethylglutaryl-coenzyme a from acetyl-coenzyme a and acetoacetyl-coenzyme a catalyzed by HMG-CoA synthase. One skilled in the art can readily determine this inhibition according to standard analytical Methods, namely Methods of enzymology (1985) 110: 19-26. Typical compounds are disclosed in, for example, US5,120,729 for β -lactam derivatives, US5,064,856 for spiro-lactone derivatives and US4,847,271 for oxetane (oxoane) compounds.

Any HMG-CoA reductase gene expression inhibitor may be used as the second compound in the combination therapy aspect of the invention. These agents may be HMG-CoA reductase transcription inhibitors that block DNA transcription, or translation inhibitors that prevent translation of mRNA encoding HMG-CoA reductase into proteins. Such inhibitors may directly affect transcription or translation, or may be bioconverted to a compound having the above-described properties by one or more enzymes in the cholesterol biosynthesis scheme, or may result in the accumulation of metabolites having the above-described activities. This effect can be readily determined by the skilled person according to standard analytical Methods, namely Methods of Enzymology (1985) 110: 9-19. Typical compounds are disclosed, for example, in US5,041,432 and e.i.mercer, prog.lip.res (1993) 32: 357 and 416.

Any CETP inhibitor may be used as the second compound in the combination therapeutic aspect of the invention. The term "CETP inhibitor" as used herein, unless otherwise indicated, refers to compounds that inhibit Cholesteryl Ester Transfer Protein (CETP) -mediated transport of various cholesteryl esters and triglycerides from HDL to LDL and VLDL. Typical compounds are disclosed in, for example, US5,512,548, j.anibiot (1996)49 (8): 815 + 816 and bioorg.med.chem.lett. (1996) 6: 1951- > 1954.

Any ACAT inhibitor may be used as the second compound in the combination therapeutic aspect of the invention. As used herein, unless otherwise indicated, the term "ACAT inhibitor" refers to an inhibitor of ACAT activity by the enzyme acyl-CoA: a compound which inhibits the lactonization of cholesterol in the diet. This inhibition can be readily determined by one skilled in the art according to standard analytical methods, namely Heider et al, Journal of Lipid Research (1983) 24: 1127. Typical compounds are disclosed, for example, in US5,510,379, WO 96/26948 and WO 96/10559.

Any squalene synthetase inhibitor can be used as the second compound in the combination therapy aspect of the invention. The term "squalene synthetase inhibitor" as used herein, unless otherwise specified, refers to a compound that inhibits the condensation reaction of two molecules of farnesyl pyrophosphate to squalene, catalyzed by the enzyme squalene synthetase. One skilled in the art can readily determine this inhibition according to standard analytical Methods, i.e., Methods of Enzymology (1985) 110: 359-373. Typical compounds are disclosed, for example, in EP-0,567,026, EP-0,645,378 and EP-0,645,377.

A therapeutically effective dose of the biphenylcarboxamide compounds of the present invention can be readily determined by those skilled in the treatment of hyperlipidemia based on the results given below. In general, it is contemplated that a therapeutically effective dose will be from about 0.001mg/kg to about 5mg/kg of the body weight of the patient to be treated, more preferably from about 0.01mg/kg to about 0.5mg/kg of the body weight of the patient to be treated. A therapeutically effective amount may also be administered in two or more doses at appropriate time intervals throughout the day. The sub-doses may be formulated in unit dosage forms, for example containing from about 0.1mg to about 350mg, more preferably from about 1 to about 200mg, of the active ingredient per unit dosage form.

Those skilled in the art know that: the exact amount and frequency of administration will depend upon the particular biphenylcarboxamide compound of formula (I) used, the particular condition being treated, the severity of the condition being treated, the age, weight and general physical condition of the particular patient, and the circumstances under which the patient is taking other drugs, including additional lipid lowering agents as described above. And the daily effective amount may be decreased or increased based on the response of the patient being treated and/or based on the evaluation of the physician prescribing the biphenylcarboxamide compounds of the instant invention. Therefore, the daily effective amounts described herein are to be referred to only.

Experimental part

The following abbreviations are used in the experimental part of the methods disclosed below:

"DMSO" means dimethylsulfoxide, "THF" means tetrahydrofuran; "DCM" means dichloromethane; "DIPE" refers to diisopropyl ether; "DMF" refers to N, N-dimethylformamide; "TFFH" refers to tetramethylfluoroformamidinium hexafluorophosphate (tetramethylfluoroformamidinium hexafluorophosphate); "NMP" means N-methyl-2-pyrrolidone; "DIPEA" refers to diisopropylethylamine; "TFA" refers to trifluoroacetic acid; and "TIS" means triisopropylsilane.

A. Synthesis of intermediates

Example A.1

a) A mixture of 4- (ethoxycarbonylmethyl) piperidine (0.0222mol) and 2-chloro-5-nitropyridine (0.0222mol) in DMSO (40ml) was placed over Na₂CO₃Is stirred in the presence of (2) for 2 hours. The reaction mixture was cooled to room temperature and poured into an ice/water mixture. The resulting precipitate was filtered and washed with water. The reaction product was purified by recrystallization from a mixture of ethyl acetate and hexane to give (5 '-nitro-3, 4, 5, 6-tetrahydro-2H- [1, 2']Bipyridyl-4-yl) -acetic acid ethyl ester (intermediate 1, melting point: 99 to 101 ℃ below zero.

b) A mixture of intermediate (1) (0.0102mol) in THF (50ml) was purified over palladium on carbon (10%; 0.3g) was hydrogenated over a catalyst at a temperature of 50 ℃ for 30 minutes. After hydrogen uptake (1 eq), the catalyst was filtered and the filtrate was evaporated to give (5 '-amino-3, 4, 5, 6-tetrahydro-2H- [1, 2' ] bipyridinyl-4-yl) -acetic acid ethyl ester (intermediate 2).

Example A.2

a) A mixture of 4- (ethoxycarbonylmethyl) hexahydropyridine (0.011mol) and 1-fluoro-4-nitrobenzene (0.011mol) in DMSO (20ml) was stirred in the presence of sodium carbonate (0.044mol) at a temperature of 60 ℃ for 2 hours. The reaction mixture was cooled to room temperature and poured into an ice/water mixture. The resulting precipitate was filtered and washed with water. The reaction product was purified by recrystallization from a mixture of ethyl acetate and hexane to give [1- (4-nitrophenyl) -hexahydropyridin-4-yl ] -ethyl acetate (intermediate 3, melting point: 83 ℃ -85 ℃ C.).

b) A mixture of intermediate (3) (0.0055mol) in THF (50ml) was purified as palladium on carbon (10%; 0.16g) was hydrogenated over a catalyst at a temperature of 50 ℃ for 30 minutes. After hydrogen uptake (1 eq), the catalyst was filtered and the filtrate was evaporated to give [1- (4-aminophenyl) -hexahydropyridin-4-yl ] -acetic acid ethyl ester (intermediate 4).

Example A.3

Thionyl chloride (3.6ml) was added to a solution of 4 '- (trifluoromethyl) - [1, 1' -biphenyl ] -2-carboxylic acid (0.025mol) in DMF (1ml) and DCM (100 ml). The mixture was stirred and refluxed for 1 hour. The solvent was evaporated. DCM (50ml) was added to the residue, which was then evaporated to give 4 '- (trifluoromethyl) - [1, 1' -biphenyl-2-carbonyl chloride (intermediate 5).

6-methyl-4 ' - (trifluoromethyl) - [1, 1 ' -biphenyl ] -2-carbonyl chloride (intermediate 6) was prepared similarly from 6-methyl-4 ' -trifluoromethyl-2-carboxylic acid using the method described above.

Example A.4

a) A mixture of Novabiochem 01-64-0261, a commercial resin (5g), benzylamine (1.765g) and titanium (IV) isopropoxide (4.686g) in DCM (50ml) was gently stirred at room temperature for 1 hour. Sodium triacetoxyborohydride (4.5g) was added and the reaction mixture was stirred at room temperature for 18 hours. Methanol (10ml) was added and the mixture was stirred for 1 hour, then filtered, washed once with DCM, once with methanol, then once with DCM (50ml) + DIPEA (5ml), first three times with DCM, then with methanol, then dried to give 5.23g of resin (I-a).

b) Piperidine-1, 4-dicarboxylic acid mono- (9H-fluoren-9-ylmethyl) ester (Fmoc-isoperidate) (0.3mmol) was dissolved in a mixture of DCM (2ml) and DMF (0.5ml) and added to a mixture of resin (I-a) (150mg) in DCM (1ml), followed by addition of TFFH (0.3mmol) in DCM (0.5ml) and DIPEA (0.6mmol) in DCM (0.5 ml). The reaction mixture was shaken at room temperature for 20 hours. The mixture was filtered and washed with DCM (3X), CH₃OH(3×)、DCM(3×)、CH₃OH(3×)、DCM(3×)、CH₃OH (3X) Wash. A mixture of piperidine in DMF (20%, 3ml) was added and the reaction mixture was shaken at room temperature for 3 hours. The mixture was filtered and washed with DCM (3X), CH₃OH(3×)、DCM(3×)、CH₃OH(3×)、DCM(3×)、CH₃OH (3X) to obtain resin (I-b).

c) A mixture of 1-fluoro-4-nitrobenzene (0.5mmol) in NMP (0.5ml) was added to resin (I-b) in NMP (3 ml). DIPEA (1mmol) dissolved in NMP (0.5ml) was added and the reaction mixture was shaken at a temperature of 50 ℃ for 18 hours. The reaction mixture was then cooled, filtered, and quenched with DCM (3X), CH₃OH(3×)、DCM(3×)、CH₃OH(3×)、DCM(3×)、CH₃OH (3X) to obtain resin (I-c).

d) A mixture of the resin (I-c) and tin chloride (2mmol) in NMP (4ml) was shaken at a temperature of 50 ℃ for 94 hours. The reaction mixture was cooled, filtered, and washed with DCM (3X), CH₃OH(3×)、DCM(3×)、CH₃OH(3×)、DCM(3×)、CH₃OH (3X) to obtain resin (I-d).

Example A.5

a) Nitropropyldialdehyde sodium hydrate (0.0143mol) and S-methylisothionium hemisulfate (0.0254mol) were dissolved in water (40ml), and hexahydropyridin-4-yl-ethyl acetate (0.0214mol) (obtained by converting hexahydropyridin-4-yl-ethyl acetate hydrochloride into its free base) was added. The reaction mixture was heated in a water bath for 10 minutes and allowed to stand overnight. The resulting precipitate was filtered and washed with water. The mother liquor layer is treated with NaHCO₃(2g) Treated and heated to 60 ℃ for 10 minutes, then the mixture was cooled and left to stand overnight. Finally, the precipitate obtained is filtered to give [1- (5-nitro-pyrimidin-2-yl) -hexahydropyridin-4-yl]Ethyl acetate (intermediate 7).

b) A solution of intermediate (7) (0.011mol) in ethyl acetate (100ml) was purified at room temperature under atmospheric pressure over palladium on carbon (10%; 0.3g) was hydrogenated over catalyst and hydrogen (3 eq) for 16 h. The reaction mixture was filtered through celite and washed with ethyl acetate. The filtrate was evaporated to give 1.9g of [1- (5-amino-pyrimidin-2-yl) -hexahydropyridin-4-yl ] -acetic acid ethyl ester (intermediate 8).

B. Synthesis of the final Compound

Example B.1

A solution of intermediate (6) (0.005mol) in dioxane (5ml) was added to a solution of intermediate (2) (0.005mol) in dioxane (15ml) and triethylamine (0.005mol) was added. The reaction mixture was stirred at room temperature for 1 hour and then diluted with water. The reaction mixture was extracted with ethyl acetate (100ml) and the organic layer was washed with brine, dried and evaporated, and then the obtained oil was purified by silica gel column chromatography using an ethyl acetate/hexane mixture as an eluent to give compound 14 (melting point: 134-.

Example B.2

4 '- (trifluoromethyl) - [1, 1' -biphenyl ] dissolved in a mixture (1ml) of DCM and DMF (80: 20)]-2-carboxylic acid (0.3mmol) was added to resin (I-d) in DCM (1 ml). A solution of TFFH (0.3mmol) in DCM (1ml) was added followed by a solution of DI PEA (0.6mmol) in DCM (1 ml). The reaction mixture was shaken for 48 hours. The reaction mixture was filtered and washed with DCM (3X), CH₃OH(3×)、DCM(3×)、CH₃OH(3×)、DCM(3×)、CH₃OH (3X) Wash. TFA/TIS/DCM (5: 2: 93) (4ml) was added and the mixture was shaken for 1 hour and then filtered. TFA/TIS/DCM (5: 2: 93) (2ml) was added further and the mixture was shaken for 15 minutes and then filtered. The filtrate was blown dry at 50 ℃ under nitrogen. The residue was taken up in DCM (3ml) and Na₂CO₃And (4) treating with an aqueous solution. The organic phase was purified by HPLC on a Chromasil 5 μm column (20 mmi.d.. times.150 mm) eluting with 100% DCM-DCM/methanol (90/10, 15 min). The desired fractions were collected and the organic solvent was evaporated to give compound (1).

Example B.3

6-methyl-4' -trifluoromethylbiphenyl-2-carboxylic acid (0.0025mol) was dissolved in anhydrous DCM (140ml) with oxalyl dichloride (2.4ml) and a few drops of DMF at 0 ℃. Then, 6-methyl-4' -trifluoromethylbiphenyl-2-carboxylic acid (0.0225mol) was added in portions in a nitrogen stream. The reaction mixture was gently heated to 40 ℃ until a homogeneous solution was produced and evolution of gas ceased. The reaction mixture was cooled to room temperature and then filtered off through a buchner funnel. The filter residue was dissolved in DCM and then added dropwise to a solution of intermediate (4) (0.025mol) and triethylamine (3g) in DCM (140ml) at 0 ℃. The reaction mixture was warmed to room temperature over a period of 90 minutes. The precipitate was filtered off, dried and purified by HPLC on Hyperprep C-18 to give compound (10).

Compound (10) (0.00042mol) was dissolved in 2-propanol (5ml) with heating. A solution of hydrochloric acid (6M) in 2-propanol (0.00042mol) was added and the mixture was cooled to room temperature, then the solvent was evaporated. The residue was crystallized from a mixture of ethanol and DI PE to give the compound (10) as a salt with an acid addition.

Compound (10) (0.00042mol) was dissolved in 2-propanol (5ml) with heating. Methanesulfonic acid (0.00042mol) was added and the solution was cooled to room temperature. The precipitate was filtered off and dried to obtain the methanesulfonic acid addition salt of compound (10).

Compound (10) (0.00042mol) was dissolved in 2-propanol (5ml) with heating. Maleic acid (0.00042mol) was added and the solution was cooled to room temperature. The precipitate was filtered off and dried to obtain a maleic acid addition salt of compound (10).

Example B.4

Compound (16) (0.0014mol) was suspended in ethanol (5ml) and NH was added₃(5ml) and the mixture was stirred under reflux overnight. The mixture was cooled to room temperature and the precipitate was filtered off. The filtrate was evaporated and purified by flash column chromatography to give compound (17).

Example B.5

4' -trifluoromethylbiphenyl-2-carboxylic acid (0.0072mol) in thionyl chloride was stirred and refluxed for 3 hours in a stream of nitrogen gas. The excess thionyl chloride was distilled off. Toluene (10ml) was added to the residue, and the mixture was evaporated in a rotary evaporator. The residue was dissolved in DCM (10ml) and cooled to 0 ℃ under a stream of nitrogen. A solution of intermediate (8) and triethylamine (1.1ml) in DCM (10ml) was added dropwise. The reaction mixture was slowly warmed to 20 ℃ and then stirred continuously for 16 hours. The solvent was evaporated, and the residue was purified by silica gel column chromatography (eluent: ethyl acetate/hexane 1: 1) to give 2.76g of compound (16).

The compounds prepared according to one of the above examples are listed in Table F-1.

TABLE F-1

C pharmacological examples

C.1.ApoB secretion quantification

HepG2 cells were cultured on 24-well plates in MEM Rega3 containing 10% fetal bovine serum. At 70% confluence, the medium was changed and test compound or vehicle (DMSO, 0.4% final concentration) was added. After 24 hours of incubation, the medium was transferred to Eppendorf vials and separated by centrifugation. Sheep antibodies against apoB were added to the supernatant and the mixture was kept at 8 ℃ for 24 hours. Rabbit anti-sheep antibodies were then added and the immune complexes were precipitated at 8 ℃ for 24 hours. The immunoprecipitates were centrifuged at 1320g for 25 minutes to pellet and pelleted with 40mM Mops, 40mM NaH₂PO₄100mM NaF, 0.2mM DTT, 5mM EDTA, 5mM EGTA, 1% Triton-X-100, 0.5% sodium Deoxycholate (DOC), 0.1% SDS, 0.2 μ M leupeptin and 0.2 μ M PMSF. The radioactivity of the pellets was quantitatively determined by liquid scintillation counting.

The resulting IC₅₀Are listed in table c.1.

TABLE C.1：pIC₅₀Value (═ logIC)₅₀Value)

Compound number	pIC
		1	7.595
2	8.219
		3	8.448
4	8.096
		5	7.416

Compound number	pIC
		6	7.934
7	8.621
		8	6.814
9	6.208
		10	7.947

Compound number	pIC
		11	7.917
12	7.503
		13	7.048
14	8.032
		15	7.591

C.1.MTP test

MTP activity was tested using a method similar to that disclosed in J.R.Wetterau and D.B.Zilversmit, Chemistry and Physics of Lipids, 38, 205-222 (1985). To prepare the donor and acceptor vesicles, the appropriate fat in chloroform was placed in a glass tube and dried under a stream of nitrogen. Containing 15mM Tris-HCl (pH7.5), 1mM EDTA, 40mM NaCl, 0.02% NaN₃(test buffer) buffer was added to the dried fat. The mixture was briefly swirled and then the fat was hydrolyzed on ice for 20 minutes. The vesicles were then prepared by sonication for a maximum of 15 minutes at room temperature (Branson 2200). Butylated hydroxytoluene was included in all vesicle preparations at a concentration of 0.1%. In a 1.5ml microcentrifuge tube, the fat transfer test mixture contained a total volume of 675 microliters of donor vesicles (40nmol phosphatidylcholine, 7.5 mol% cardiolipin, and 0.25 mol% glycerotris [1-¹⁴C]Oleate), receptor vesicles (240nmol phosphatidylcholine) and 5mg BSA. Test compounds were added dissolved in DMSO (0.13% final concentration). After a 5 min pre-incubation at 37 ℃, the reaction was started by adding MTP in 100 μ l dialysis buffer. By adding 1mM EDTA, 0.02% NaN at 15mM Tris-HClpH7.5₃The reaction was terminated by pre-equilibration of 400. mu.l DEAE-52 cellulose in (1: 1, vol/vol). The reaction mixture was shaken in an Eppendorf centrifuge at maximum rate for 4 minutes and centrifuged for 2 minutes (4 ℃) to pellet the DEAE-52 bound donor vesicles. The supernatant containing the acceptor liposome in the same portion was counted and used¹⁴C]Counting to calculate the percentage of triglycerides transferred from the donor into the acceptor vesicles.

Claims (7)

1.A compound of formula (I)

The N-oxides, the pharmaceutically acceptable acid addition salts and the stereochemically isomeric forms thereof,

wherein

R¹Is hydrogen, C_1-4Alkyl, halogen, or polyhaloC_1-4An alkyl group;

R²is hydrogen, C_1-4Alkyl, halogen, or polyhaloC_1-4An alkyl group;

R³is hydrogen or C_1-4An alkyl group;

R⁴is hydrogen, C_1-4Alkyl, or halogen;

n is an integer of 0 or 1;

X¹and X²Or both are carbon;

X³is carbon;

y is 0 or NR⁶Wherein R is⁶Is hydrogen or C_1-4An alkyl group; and

R⁵is hydrogen; optionally is covered with C_1-4Alkoxy, cyano, polyhaloC_1-4Alkyl or aryl substituted C_1-6An alkyl group; c optionally substituted by aryl_2-6An alkenyl group; c optionally substituted by aryl_3-6An alkynyl group;

aryl or heteroaryl;

aryl is phenyl; is one, two or three of the substituents are independently selected from nitro, azido, cyano, halogen, hydroxyl, C_1-6Alkyl radical, C_3-6Cycloalkyl radical, C_1-4Alkoxy, polyhalo C_1-6Alkyl, amino, mono-or di (C)_1-6Alkyl) phenyl substituted with a substituent of the amino group; heteroaryl is pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, triazinyl, triazolyl,

Imidazolyl, pyrazolyl, thiazolyl, isothiazolyl, oxazolyl, pyrrolyl, furanyl or thienyl; and optionally substituted by one, two or three substituents each independently selected from nitro, or a combination thereof,

Azido, cyano, halogen, hydroxy, C_1-6Alkyl radical, C_3-6Cycloalkyl radical, C_1-4Alkoxy, polyhalo C_1-4Alkyl, amino, mono-or di (C)_1-6Alkyl) amino.

2. The compound of claim 1, wherein n is the integer 0.

3. The compound of claim 1, wherein n is the integer 1.

4. A pharmaceutical composition comprising a pharmaceutically acceptable carrier and a therapeutically active amount of a compound as claimed in any one of claims 1 to 3.

5. A process for preparing a pharmaceutical composition as claimed in claim 4, wherein a therapeutically active amount of a compound as claimed in any one of claims 1 to 3 is intimately mixed with a pharmaceutically acceptable carrier.

6. A compound according to any one of claims 1 to 3 for use as a medicament.

7. A process for the preparation of a compound of formula (I), wherein

a) Reacting an intermediate of formula (II) wherein R³、R⁴、R⁵、n、Y、X¹、X²And X³As defined in claim 1, in the same manner,

with a biphenylcarboxylic acid or halide of the formula (III), in which R¹And R²Is as defined in formula (I) and Q¹Selected from hydroxyl and halogen, in at least one reaction-inert solvent and optionally in the presence of a suitable base,

b) or, the compounds of formula (I) are converted into each other according to conversion reactions of the prior art; or, if desired, converting a compound of formula (I) into an acid addition salt, or conversely, converting an acid addition salt of a compound of formula (I) into its free base form with a base; and, if desired, preparing stereochemically isomeric forms thereof.