OA12363A - Therapeutic combination of a CETP inhibitor and atTartrate salts of thiazolidinedione derivative. orvastatin. - Google Patents

Description

012363 -4-

THERAPEUTIC COMBINATION

This invention relates to pharmaceutical combinations of cholestérol ester transfer protein (CETP) inhibitors in particular, [2R, 4S]4-[(3,5-bis-trifluoromethyi- benzyl)-methoxycarbonyl-aminoJ-2-ethyl-6-trifluoromethyl-3.4-dihydro-2H-quinoline-1- carboxylic acid ethyl ester, and atorvastatin and métabolites thereof and pharmaceutically acceptable salts thereof.

BACKGROUND OF THE INVENTION

[2R, 4S]4-[(3,5-Bis-trifluoromethyl-benzyl)-methoxycarbonyl-amino3-2-ethyl-6-trifluoromethyl-3.4-dihydro-2H-quinoline-1-carboxylic acid ethyl ester is disclosed inPCT/IB99/01532 application published as WO 00/17164 on March 30, 2000 as aCETP inhibitor for the élévation of certain plasma lipid levels and to lower certainother plasma lipid levels and accordingly to prevent the occurrence of and treatdiseases such as lipid abnormalities, atherosclerosis and cardiovascular diseases. ..That published application also discloses the combination of a genus of 4-carboxyamino-2-substituted-1,2,3,4-tetrahydroquinolines with a preferred group of 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase inhibitors beinglovastatin, simvastatin, pravastatin, fluvastatin, atorvastatin or rivastatin.

Commonly assigned U.S. provisional application serial no. 60/168,051 filedNovember 30, 1999 discloses crystalline forms of [2R, 4S]4-[(3,5-bis-trifluoromethyl-benzyl)-methoxycarbonyI-amino}-2-ethyl-6-trifiuQromethyl-3,4-dÎhydro-2H-quinolÎne-1-carboxylic acid ethyl ester, specifically anhydrous and monoethanolate crystallineforms.

Commonly assigned U.S. provisional application serial no. ^β/167,967 filedNovember 30, 1999 discloses methods for making [2R, 4S]4-[(3,5-bis-trifluoromethyl-benzyl)-methoxycarbonyl-aminoj-2-ethyl-6-trifluoromethyl-3,4-dihydro-2H-quinoIine-1-carboxylic acid ethyl ester.

The conversion of 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) tomevalonate is an early and rate-limiting step in the cholestérol biosynthetic pathway.This step is catalyzed by the enzyme HMG-CoA reductase. Statiris inhibit HMG-CoAreductase from catalyzing this conversion. As such, statins are lipid lowering agents.

Atorvastatin calcium, disclosed in U.S. Patent No. 5,273,995, which isincorporated herein by reference, is currently sold as Lipitor® and has the formula 012363

Atorvastatin calcium is a sélective, compétitive inhibitor of HMG-CoA. As such,atorvastatin calcium is a potent lipid lowering compound. The free carboxylic acidform of atorvastatin existe predominantly as the lactone of the formula

and is disclosed in U.S. Patent No. 4,681,893, which is incorporated herein byréférencé.

Hydroxylated dérivatives of atorvastatin (hydroxy métabolites) having the10 formula below wherein R1 is hydroxy are disclosed in U.S. Pat. No. 5,385,929, the disclosure of which is hereby incorporated by référencé. ♦ 012363 -3-

One dérivative disclosed in U.S. Pat No. 5,385,929 is (2R-trans)-5-(4-fluorophenyl)-2-(1-methylethyl)-N-(2-hydroxyphenyl)-4-phenyl-1-[2-(tetrahydro-4- 5 hydroxy-6-oxo-2H-pyran-2-yl)ethyl]-1 H-pyrrole-3-carboxamide.

Another dérivative disclosed in U.S. Pat. No. 5,385,929 (Exemple 2) is (2R- trans)-5-(4-fluorophenyl)-2-(1-methylethyl)-N-(3-hydroxyphenyl)-4-phenyl-1-{2- (tetrahydro-4-hydroxy-6-oxo-2H-pyran-2-yl)ethyI]-1H-pyrrole-3-carboxamide.

Yet another dérivative disclosed in U.S. Pat. No. 5,385,929 (Example 1) is 10 (2R-trans)-5-(4-fluorophenyl}-2-(1 -methy!ethyl)-N-(4-hydroxyphenyl)-4-phenyl-1 -[2-(tetrahydro-4-hydroxy-6-oxo-2H-pyran-2-yl)ethyl]-1H-pyrrole-3-carboxamide.

Atherosclerosis is a condition characterized by irregularly distributed lipiddeposits in the intima of arteries, including coronary, carotid and peripheral arteries.Atherosclerotic coronary heart disease (hereinafter termed “CHD”) accounts for 53% 15 of ail deaths attributable to a cardiovascular event. CHD accounts for nearly one-half(about $50-60 billion) of the total U.S. cardiovascular healthcare expenditures andabout 6% of the overall national medical bill each year. Despite attempts to modifysecondary risk factors such as, inter alia, smoking, obesity and lack of exercise, andtreatment of dyslipidemia with dietary modification and drug therapy, CHD remains 20 the most common cause of death in the United States. 012363 -4-

Risk for development of this condition has been shown to be stronglycorrelated with certain plasma lipid levels. While elevated LDL-C may be the mostrecognized form of dysfipidemia, it is by no means the only significant lipid associatedcontributor to CHD. Low HDL-C is also a known risk factor for CHD (Gordon, D.J., etal.,: “High-density Lipoprotein Cholestérol and Cardiovascular Disease”, Circulation,(1989), 79: 8-15).

High LDL-cholesterol and triglycéride levels are positively correlated, whilehigh levels of HDL-cholesterol are negatively correlated with the risk for developingcardiovascular diseases. Thus, dyslipidemia is not a unitary risk profile for CHD butmay be comprised of one or more lipid aberrations.

Among the many factors controlling plasma levels of these diseasedépendent principes, cholesteryl ester transfer protein (CETP) activity affects ailthree. The rôle of this 70,000 dalton plasma glycoprotein found in a number of animalspecies, including humans, is to transfer cholesteryl ester and triglycéride betweenlipoprotein particles, including high density lipoproteins (HDL), low density lipoproteins(LDL), very low density lipoproteins (VLDL), and chylomicrons. The net resuit ofCETP activity is a lowering of HDL cholestérol and an increase in LDL cholestérol.

This effect on lipoprotein profile is believed to be pro-atherogenic, especially insubjects whose lipid profile constitues an increased risk for CHD.

No whotly satisfactory HDL-elevating thérapies exist. Niacin can significantlyincrease HDL, but has serious toleration issues whïch reduce compliance. Fibratesand the HMG CoA reductase inhibitors raise HDL-C only modestly (-10-12%). As aresuit, there is a significant unmet medical need for a well-tolerated agent which cansignificantly elevate plasma HDL levels, thereby reversing or slowing the progressionof atherosclerosis.

High levels of blood cholestérol and blood lipids are conditions involved in theonset of atherosclerosis. It is well known that inhibitors of 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMG-CoA reductase) are effective in lowering the level ofblood plasma cholestérol, especially low density lipoprotein cholestérol (LDL-C), inman (Brown and Goldstein, New England Journal of Medicine, 1981, 305, No. 9, 515-517). It has now been established that lowering LDL-C levels affords protection fromcoronary heart disease (see, e.g., The Scandinavian Simvastatin Survival StudyGroup: Randomised trial of cholestérol lowering in 4444 patients with coronary heartdisease: the Scandinavian Simvastatin Survival Study (4S), Lancet, 1994, 344, 1383- .012363 -5- 89; and Shepherd, J. et al., Prévention of coronary heart disease with pravastatin inmen with hypercholesterolemia, New England Journal of Medicine, 1995, 333, 1301-07).

Angina pectoris is a severe constricting pain in the chest, often radiating fromthe précordium to the left shoulder and down the left arm. Often angina pectoris isdue to ischemia of the heart and is usually caused by coronary disease.

Currently the treatment of symptomatic angina pectoris varies significantlyfrom country to country. In the U.S., patients who présent with symptomatic, stableangina pectoris are frequently treated with surgical procedures or PTCA. Patientswho undergo PTCA or other surgical procedures designed to treat angina pectorisfrequently expérience complications such as restenosis. This restenosis may bemanifested either as a short term proliférative response to angioplasty-inducedtrauma or as long term progression of the atherosclerotic process in both graftvessels and angioplastied segments.

The symptomatic management of angina pectoris involves the use of anumber of drugs, frequently as a combination of two or more of the following classes:beta blockers, nitrates and calcium channel blockers. Most, if not ail, of these patientsrequire therapy with a lipid lowering agent as well. The National CholestérolEducation Program (NCEP) recognizes patients with existing coronary artery diseaseas a spécial class requiring aggressive management of raised LDL-C.

SUMMARY OF THE INVENTION

This invention is directed to a pharmaceutical composition comprising atherapeutically effective amount of a composition comprising: a. [2R, 4S]4-[(3,5-bis-trifluoromethyl-benzyl)-methoxycarbonyl-amino]-2-ethyl-5-trifluoromethyl-3,4-dihydro-2H-quinoline-1-carboxylic acid ethyl ester; b. atorvastatin or the corresponding cyclized iactone form ofatorvastatin, a 2-hydroxy, 3-hydroxy or 4-hydroxy dérivative of atorvastatin or thecyclized Iactone form of atorvastatin, or a pharmaceuticaîly acceptable sait thereof;and c. a pharmaceuticaîly acceptable carrier, vehicle or diluent.

As used herein the dérivatives (hydroxy métabolites) of the cyclized Iactone form of atorvastatin or atorvastatin (the open chain form) described as 2-hydroxy, 3- hydroxy or 4-hydroxy hâve the Formula I and IA below, respectively, 012363

OH

\ /CH2 , , ΝΗ

Formula and

wherein R1 is hydroxy. 012363 t -7-

Preferably the composition comprises atorvastatin and it is especially preferred that the composition comprises the hemicalcium sait oî atorvastatin.

Preferably R1 is 2-hydroxy.

This invention is also directed to a method for treating a mammal (e.g., ahuman either maie or female) in need of therapeutic treatment comprisingadministering to said mammal a therapeutically effective amount of: (a) a first compound, said First compound being [2R, 4S]4-[(3,5-bis-trifluoromethyl-benzyl)-methoxycarbonyl-amino3-2-ethyl-6-trifluoromethyl-3,4-dihydro-2H-quinoline-1-carboxylic acid ethyl ester; and (b) a second compound, said second compound being atorvastatin orthe cyclized lactone form of atorvastatin, a 2-hydroxy, 3-hydroxy or 4-hydroxydérivative of said compounds or a pharmaceutically acceptable sait thereof;wherein said first compound and said second compound are each optionally andindependently administered together with a pharmaceutically acceptable carrier,vehicle or diluent.

Preferably the composition comprises atorvastatin and it is especiallypreferred that the composition comprises the hemicalcium sait of atorvastatin.

Preferably R1 is 2-hydroxy.

Preferably the first compound and the second compound are administeredsimultaneously.

Preferably the first compound and the second compound are administeredsequentially in either order.

Preferably the therapeutic treatment comprises antiatherosclerotic treatment.Preferably the therapeutic treatment comprises slowing and/or arresting the progression of atherosclerotic plaques.

Preferably the progression of atherosclerotic plaques is slowed in coronary arteries.

Preferably the progression of atherosclerotic plaques is slowed in carotidarteries.

Preferably the progression of atherosclerotic plaques is slowed in theperipheral arterial System.

Preferably. the treatment of atherosclerosis causes the régression ofatherosclerotic plaques. 012363 -8-

Preferably the régression of atherosclerotic plaques occurs in coronary arteries.

Preferably the régression of atherosclerotic plaques occurs in càrotid arteries.Preferably the régression of atherosclerotic plaques occurs in the peripheral arterial System.

Preferably the therapeutic treatment comprises HDL élévation treatment andantihyperlipidemic treatment (including LDL lowering).

Preferalby the therapeutic treatment comprises antianginal treatmentPreferably the therapeutic treatment comprises cardiac risk management.

This invention is also directed to a kit for achieving a therapeutic effect in a mammal comprising a therapeutically effective amount of a composition comprising: a. [2R, 4S]4-[(3,5-bis-trifluoromethyl-benzyl)-methoxycarbonyl-amino]-2-ethyl-6-trifluoromethyl-3,4-dihydro-2H-quinoline-1-carboxylic acid ethyl esterand a pharmaceutically acceptable carrier, vehicle or diluent in a first unit dosageform; b. atorvastatin or the cyclized lactone form of atorvastatin, a 2-hydroxy, 3-hydroxy or 4-hydroxy dérivative of said compounds or a pharmaceuticallyacceptable sait thereof and a pharmaceutically acceptable carrier, vehicle or diluentin a second unit dosage form; and c. means for containing said first and second dosage forms.Preferably the composition comprises atorvastatin and it is especially preferred that the composition comprises the hemicaicium sait of atorvastatin.Preferably R1 is 2-hydroxy.

This invention is also particulariy directed to a kit wherein the therapeuticeffect is the prévention and/or treatment of atherosclerosis.

This invention is still more particulariy directed to a kit wherein the treatmentof atherosclerosis slows the progression of atherosclerotic plaques.

This invention is further directed to a kit wherein the progression ofatherosclerotic plaques is slowed in coronary arteries.

This invention is still further directed to a kit wherein the progression ofatherosclerotic plaques is slowed in carotid arteries.

This invention is still further directed to a kit wherein the progression ofatherosclerotic plaques is slowed in the peripheral arterial System. 012363 -9-

This invention is still further directed to a kit wherein the treatment of atherosclerosis causes the régression of atherosclerotic plaques.

This invention is still further directed to a kit wherein the régression of atherosclerotic plaques occurs in coronary arteries.

This invention is still further directed to a kit wherein the régression ofatherosclerotic plaques occurs in carotid arteries.

This invention is still further directed to a kit wherein the régression ofatherosclerotic plaques occurs in the peripheral arterial System.

This invention is still more particularly directed to a kit wherein the therapeuticeffect is treatment of low HDL levels and hyperlipidemia.

This invention is still more particularly directed to a kit wherein the therapeuticeffect is the prévention and/or treatment of angina pectoris.

This invention is also particularly directed to a kit wherein the therapeuticeffect is the management of cardiac risk.

This invention is also directed to a first pharmaceutical composition for usewith a second pharmaceutical composition for achieving a therapeutic effect in amammal, which effect is greater than the individual therapeutic effects achieved byadministering said first or second pharmaceutical compositions separately and whichsecond pharmaceutical composition comprises an amount of atorvastatin or thecyclized lactone form of atorvastatin, a 2-hydroxy, 3-hydroxy or 4-hydroxy dérivativeof said compounds or a pharmaceutically acceptable sait thereof and apharmaceutically acceptable carrier, vehicle or diluent, said first pharmaceuticalcomposition comprising of [2R, 4S]4-[(3,5-bis-trifluoromethyl-benzyl)-methoxycarbonyl-amino]-2-ethyl-6-trifluoromethyl-3,4-dihydro-2H-quinoiine-1-carboxylic acid ethyl ester and a pharmaceutically acceptable carrier, vehicle ordiluent.

This invention is also directed to a first pharmaceutical composition for usewith a second pharmaceutical composition for achieving a therapeutic effect in amammal, which effect is greater than the individual therapeutic effects achieved byadministering said first or second pharmaceutical compositions separately and whichsecond pharmaceutical composition comprises an amount of [2R,4S]4-[(3,5-bis-trifluoromethyl-benzyl)-methoxycarbonyl-amino]-2-ethyl-6-trifluoromethyl-3,4-dihydro-2H-quinoline-1 -carboxylic acid ethyl ester and a pharmaceutically acceptable carrier,vehicle or diluent, said first pharmaceutical composition comprising an amount of 012363 -10- atorvastatin or the cyclized lactone form of aton/astatin, a 2-hydroxy, 3 hydroxy or 4-hydroxy dérivative of said compounds or a pharmaceutically acceptable sait thereofand a pharmaceutically acceptable carrier, vehicle or diluent.

In the above two pharmaceutical compositions the following are preferredembodiments.

Preferably the therapeutic effect is the prévention and/or treatment ofatherosclerosis.

Preferably the therapeutic effect is a LDL-C lowering effect and a HDL-Craising effect in a mammal suffering from hyperiipidemia and low HDL levels.

Preferably the therapeutic effect is the prévention of the occurrence of anginain a mammal at high risk thereof.

Preferably the therapeutic effect is the management of cardiac risk in amammal at risk of suffering an adverse cardiac event.

Preferably the composition comprises atorvastatin and it is especiallypreferred that the composition comprises the hemicalcium sait of atorvastatin.

Preferably R1 is 2-hydroxy.

Preferably the antiatherosclerotic effect is manifested by a slowing of theprogression of atherosclerotic plaques.

Preferably the progression of atherosclerotic plaques is slowed in coronaryarteries.

Preferably the progression of atherosclerotic plaques is slowed in carotidarteries.

Preferably the progression of atherosclerotic plaques is slowed in theperipherai arterial System.

Preferably the antiatherosclerotic effect is manifested by a régression ofatherosclerotic plaques.

Preferably the régression of atherosclerotic plaques occurs in coronaryarteries.

Preferably the régression of atherosclerotic plaques occurs in carotid arteries.

Preferably the régression of atherosclerotic plaques occurs in the peripherai arterial System.

The expression "pharmaceutically-acceptable sait" refers to nontoxic anionic salts containing anions such as (but not limited to) chloride, bromide, iodide, sulfate, bisulfate, phosphate, acetate, maleate, fumarate, oxalate, lactate, tartrate, citrate, «r 012363 -11- gluconate, methanesulfonate and 4-toluene-sulfonate. The expression also refers tonontoxic cationic salts such as (but not limited to) sodium, potassium, calcium,magnésium, ammonium or protonated benzathine (N.N'-dibenzyiethyienediamine),choline, ethanoiamine, diethanolamine, ethylenediamine, meglamine (N-methyl-glucamine), benethamine (N-benzylphenethylamine), piperazine or tromethamine (2-amino-2-hydroxymethyl-1,3-propanediol).

Where used herein, the term “cardiac risk" means the iikelihood that a subjectwill suffer a future adverse cardiac event such as, e.g., myocardial infarction, cardiacarrest, cardiac failure or cardiac ischaemia. Cardiac risk is calculated using theFramingham Risk Equation. The term “cardiac risk management” means that the riskof future adverse cardiac events is substantially reduced.

As used herein, the expressions "reaction-inert solvent" and "inert solvent"refers to a solvent or a mixture thereof which does not interact with starting matériels,reagents, intermediates or products in a manner which adversely affects the yield ofthe desired product. A chemist of ordinary skill will recognize that certain compounds of thisinvention will contain one or more atoms which may be in a particular stereochemicalor géométrie configuration, giving rise to stereoisomers and configurational isomers.Ail such isomers and mixtures thereof are included in this invention. Hydrates andsolvatés of the compounds of this invention are also included.

As used herein the term mammals is meant to refer to ail mammals whichcontain CETP in their plasma, for example, rabbits and primates such as monkeysand humans (e.g., male orfemale). Certain other mammals e.g., dogs, cats, cattle,goats, sheep and horses do not contain CETP in their plasma and so are notincluded herein.

The term "treating", "treat" or "treatment" as used herein includes preventative(e.g., prophylactic) and palliative treatment.

By "pharmaceutically acceptable" is meant the vehicle, carrier, diluent,excipients, and/or sait must be compatible with the other ingrédients of theformulation, and not deleterious to the récipient thereof. ♦ 0i2363 -12- DETAILED DESCRIPTION OF THE INVENTION[2R, 4S]4-[(3,5-Bis-trifluoromethyl-benzyl)-methoxycarbonyl-amino]-2-ethyl-6- trifluoromethyl-3.4-dihydro-2H-quinoline-1-carboxyiic acid ethyl ester is disclosed inPCT/IB99/01532 application published as WO 00/17164 on March 30, 2000 and may 5 readily be prepared as described therein (see Examples 7 (racemate) and Example120). Methods for préparation of this compound (and polymorphs thereof) are alsodisclosed in commonly assigned U.S. provisional applications serial nos. 60/168,051and 60/168,051 and hereinafter.

Example 1 10 c/s^-fQ.S-Bis-trifluoromethvl-benzvD-methoxvcarbonvI-aminol^-ethvl-e- trifluoromethyl-3.4-dihvdro-2H-quinoline-1-carboxvlic acid ethvl ester: A solution of c/s-4-(3,5-bis-trifluoromethyl-benzyIamino)-2-ethyl-6-trifiuoromethyl-3,4-dihydro-2K-quinoline-1-carboxylic acid ethyl ester (2.0 g, 3.7 mmol) and pyridine(0.58 g, 7.4 mmol) in 100 mL of dichloromethane was cooled in an ice/water bath as 15 methyl chloroformate (0.87 g, 9.2 mmol) was added slowly. After stirring overnight at room température, the reaction mixture was washed twice with a 2N hydrochloric acidsolution, dried over magnésium sulfate, filtered and concentrated in vacuo to affordthe crude product, which was purified by silica gel chromatography using 5-10% ethylacetate/hexanes as eluent to afford 1.8 g of the title product. MS mfz 601 (M+ +1); 20 1H NMR (coalescing mixture of conformers, CDCI3) δ 0.6-0.8 (bm, 3H), 1.2-1.3 (bm,3H), 1.3-1.5 (bm, 2H), 1.6-1.75 (bm, 1H), 2.1-2.3 (bm, 1H), 3.7-3.9 (bs, 3H), 4.0-4.4(bm, 4H), 5.0-5.6 (bm, 2H), 7.1 (s, 1H), 7.4-7.6 (bm, 2H), 7.6-7.8 (bm, 3H).

[2R,4S]4-[(3,5-bis-trifluoromethyl-benzyl)-methoxycarbonyl-amino]-2-ethyl-6- 25 trifluoromethyl-3,4-dihydro-2H-quinoline-1-carboxyl»c acid ethyl ester was prepared inoptically enriched form by resolution of the corresponding racemate, or anintermediate in its synthesis, using standard methods.

Exampie 2 ( 1 -Benzotriazol-1 -vl-propvl|-(4-trifluorornethvl-phenvi)-amine 30 A two liter, four neck flask under nitrogen atmosphère was charged with benzotriazole (36.96 g, 310 mmol, 1.0 equiv) and dry toluene (400 mL). A room température solution of 4-(trifluoromethyl)aniline (39.1 mL, 310 mmol, 1.0 equiv) and 50 mL toluene was added over one minute. A room température solution of propionaldéhyde (24.6 mL, 341 mmol, 1.1 equiv) and 50 mL toluene was then added ,.- 0)2363 -13- over 20 minutes. There was an exotherm from 23°C to 30°C during this addition.After stirring 24 h, n-heptane (500 mL) was added, and the sîurry stirred an additional1 h. The suspension was filtered, the soiids were washed with n-heptane (1 x 100mL, then 1 x 200 mL, and dried. (1-Benzotriazol-1-yl-propyl)-(4-trifiuoromethyl- 5 phenyi)-amine was isolated as shiny white needles (81.3 g, 82%). After 24 h, asecond crop was isolated from the filtrate (8.7 g, 9%). mp 130-132 °C; 1H NMR(DMSO-d6,400 MHz) δ 0.82 (t, 3H, J=7.5 Hz), 2.25 (m, 2H), 6.49 (m, 1H), 6.80 (d, 2H, J=8.7 Hz), 7.35 (m, 3H), 7.50 (m, 1H), 7.88 (d, 1H, J=8.3 Hz), 7.99 (m, 1H), 8.09(d, 1H, J=8.5 Hz); 13C NMR (DMSO-d6, 100 MHz) δ 149.32, 146.19, 131.46, 127.73, 10 126.8,125.33 (q, J=270 Hz), 124.44,119.88,118.27 (q, J=31.7 Hz), 112.91,111.56, 71.03, 28.08, 10.29; DEPT spectrum: quaternary carbons δ 149.32,146.19,131.46,125.33, 118.27; CH carbons δ 127.73, 126.8, 124.44, 119.88, 112.91, 111.56, 71.03;CH2 carbon δ 28.08; CH3 carbon δ 10.29; IR (drifts) 3292 (s), 3038 (m), 2975 (m),1621 (s), 1331 (s), 1320 (s), 1114 (vs); Anal. Calcd for C16H1SN4F3: C, 59.99; H, 4.72; 15 N, 17.49. Found (first crop): C, 60.16; H, 4.74; N, 17.86. Found (second crop): C,59.97; H, 4.66; N, 17.63.

Exampie 3 cis-(2-Ethyl-6-trifluoromethyl-1,2,3.4-tetrahydro-quinolin-4-vl)-carbamic acid benzvl ester 20 A one iiter, four neck flask under nitrogen atmosphère was charged with N-vinyl-carbamic acid benzyl ester (27.66 g, 156 mmol, 1.0 equiv) and dry toluene (500 mL).(1-Benzotriazol-1-yl-propyl)-(4-trifluoromethyl-phenyl)-amine (50.0 g, 156 mmol, 1.0equiv) and p-to!uenesuifonic acid monohydrate (297 mg, 1.56 mmol, 0.01 equiv) wereadded, and the mixture heated to 70°C. After 2 h, the mixture was cooled to room 25 température and transfered to a separatory funnel. Ethyl acetate (500 mL) was

added. The mixture was washed 1 x 200 mL 1N NaOH, 1 x 200 mL H20,1 x 200 mLbrine, and dried (MgSO4). The mixture was filtered and the soiids washed 1 x 50 mLethyl acetate. The filtrate was concentrated to approximately 250 mL. 500 mLtoluene were added, and the mixture concentrated to approximately 500 mL. 500 mL 30 n-heptane were added, the slurry was stirred 1 h, filtered through a Buchner funnel, and dried. cis-(2-Ethyl-6-trif!uoromethyl-1,2,3,4-tetrahydro-quinolin-4-yl)-carbamic

acid benzyl ester was isolated as a white powder (45.04 g, 76%): mp 155-157 °C; 1H NMR (DMSO-d6, 400 MHz) δ 0.92 (t, 3H, J=7.5 Hz), 1.5 (m, 3H), 2.00 (m, 1H), 3.35 , o’23B3 -14- (m, 1H), 4.77 (m, 1H), 5.07 (d, 1H, J=12.5 Hz), 5.15 (d, 1H, J=12.5 Hz), 6.35 (s, 1H),6.61 (d, 1H, J=8.5 Hz), 7.12 (s, 1H), 7.18 (dd, 1H, J=1.9, 8.5 Hz), 7.4 (m, 5H), 7.70(d, 1H, J=9.1 Hz); 13C NMR (DMSO-d6,100 MHz) δ 157.03, 149.02, 137.79,128.82,128.23,128.03,125.9 (q, J=270 Hz), 125.06, 123.50,121.73, 115.2 (q, J=31.7 Hz),113.33, 65.85, 52.09, 47.83, 34.02, 28.68, 9.93; DEPT spectrum: quaternarycarbons δ 157.03,149.02,137.79,125.9,121.73, 115.2; CH carbons δ 128.82,128.23, 128.03, 125.06,123.50, 113.33, 52.09, 47.83; CH2 carbons δ 65.85, 34.02,28.68; CH3 carbon δ 9.93; IR (drifts) 3430 (m), 3303 (s), 2951 (m), 1686 (vs), 1542(vs), 1088 (vs); MS (APCI+) m/z (rel. intensity) 379 (M+H+, 53), 228 (100); Anal.

Calcd for C20H21N2O2F3: C, 63.48; H, 5.59; N, 7.40; Found: C, 63.69; H, 6.06, N,7.36.

Example 4 cis-4-Benzvloxvcarbonvlamino-2-ethvl-6-trifluoromethvl-3,4-dihvdro-2H-quinoline-1- carboxvlic acid ethvl ester A three liter, four neck flask under nitrogen atmosphère was charged with cis-(2-ethyl-6-trifluoromethyl-1,2,3,4-tetrahydro-quinolin-4-yl)-carbamic acid benzyl ester(96.0 g, 254 mmol, 1.0 equiv), dry dichloromethane (720 mL), and dry pyridine (103mL, 1.27 mol, 5.0 equiv). A solution of ethyl chioroformate (121 mL, 1.27 mol, 5.0equiv), in dry dichloromethane (240 mL), was added slowly over 4 h. The additionwas exothermic and required a reflux condenser. Once the chioroformate additionwas complété, the reaction was cooled in an ice bath and 1350 mL 1N NaOH wereadded. The mixture was stirred 15 min, then transferred to a separatory funnel. Theiayers were separated and the aqueous extracted 1 x 1L dichloromethane. Thecombined dichloromethane Iayers were washed 1 x 1350 mL 1N HCl, 1 x 1Lsaturated aq. NaHCO3, 1 x 1L brine, and dried (Na2SO4). The mixture was filtered,and the filtrate concentrated to an orange oil. 570 mL abs. éthanol were added, andthe solution was concentrated. The solids were dissolved in 1370 mL abs. éthanol.570 mL H2O were added dropwise over 45 min. The résultant thick siurry was stirred18 h and filtered. The solids were washed with cold 7:3 abs. ethanol/water (1 x 250mL, then 1 x 100 mL) and dried (vac oven, 45°C) to give cîs-4- benzyloxycarbonylamino-2-ethyl-6-trifluoromethyl-3,4-dihydro-2H-quinoline-1- carboxylic acid ethyl ester as a white, crystalline solid (94.54 g, 83%): mp 92-96°C; 1H NMR (CDCIs, 400 MHz) δ 0.84 (t, 3H, J=7.4 Hz), 1.28 (t, 3H, J=7.0 Hz), 1.4 (m, «.θ 12363 -15- 2Η), 1.62 (m, 1 Η). 2.53 (m, 1Η), 4.23 (m, 2H), 4.47 (m, 1H), 4.79 (m, 1H), 5.01 (ci, 1H, J=9.2 Hz), 5.18 (m, 2H), 7.4 (m, 5H), 7.5 (m, 2H), 7.57 (m, 1H); 13C NMR (CDCI3,100 MHz) δ 155.97, 154.43,139.44, 136.21, 134.33, 128.61, 128.33, 128.22, 126.32(q, J=31.7 Hz), 126.18,124.22,124.19, 124.12 (q, J=273 Hz), 120.74, 120.70, 67.22,62.24, 53.47, 46.79, 37.75,28.25, 14.38, 9.78; DEPT spectrum: quaternary carbonsδ 155.97,154.43, 139.44,136.21, 134.33,126.32, 124.12; CH carbons δ 128.61, 128.33.128.22, 126.18,124.22, 124.19,120.74, 120.70, 53.47, 46.79; CH2 carbons δ 67.22, 62.24, 37.75, 28.25; CH3 carbons δ 14.38, 9.78; IR (drifts) 3304 (s), 3067 (m),3033 (m), 2982 (m), 2932 (m), 1723 (s), 1693 (s), 1545 (s); MS (APCI+) m/z (rel.intensity) 451 (M+H\ 2), 300 (100); Anal. Calcd for C23H2SN2O4F3: C, 61.33; H, 5.60;N, 6.22. Found: C, 61.07; H, 5.69; N, 6.22.

Example 5 cis-4-Amino-2-ethvl-6-trifluoromethyl-3.4-dihvdro-2H-quinoline-1 -carboxvlic acid ethvl ester A one liter, four neck flask under nitrogen atmosphère was charged with cis-4-benzyloxycarbonylamino-2-ethyl-6-trifluoromethy!-3,4-dihydro-2H-quinoline-1-carboxylic acid ethyl ester (40.1 g, 89 mmol, 1.0 equiv), methanol (400 mL), andammonium formate (14.0 g, 223 mmol, 2.5 equiv). 10% Pd/C, 50% water wet (4.0 g)was added, and the slurry heated to 40° C over 1 h. After 1.5 h, the mixture wascooled to room température and filtered through Celite®. The cake was washed 2 x100 mL methanol. The filtrate was concentrated to approximately 75 mL, transferredto a separatory funnel, and diluted with 400 mL ethyl acetate. The mixture waswashed 1 x 125 mL saturated aq. NaHCO3,1 x 100 mL brine, and dried (Na2SO4).The mixture was filtered and the filtrate concentrated to a clearoil. The oil wascrystallized from 100 mL n-heptane to give cis-4-amino-2-ethyl-6-trifluoromethyl-3,4-dihydro-2H-quinoline-1-carboxylic acid ethyl ester as a white crystalline soiid (26.05 g,93%): mp 61.5-63.5° C; 1H NMR (CDCi3,400 MHz) δ 0.79 (t, 3H, J=7.5 Hz), 1.24 (m,4H), 1.42 (m, 1H), 1.51 (brs, 2H), 1.62 (m, 1H), 2.46 (m, 1H), 3.73 (m, 1H), 4.17 (m,2H), 4.36 (m, 1H), 7.44 (m, 2H), 7.66 (m, 1H); 13C NMR (CDCJ3, 100 MHz) δ 154.6,139.3,138.9, 126.3 (q, J=32 Hz), 125.7, 124.3 (q, J=271 Hz), 123.5, 119.8, 61.96,54.16,46.91, 41.50, 28.85,14.38, 9.60; DEPT spectrum: quaternary carbons δ154.6,139.3, 138.9, 126.3,124.3; CH carbons δ 125.7, 123.5,119.8, 54.16, 46.91;CH2 carbons δ 61.96, 41.50, 28.85; CH3 carbons δ 14.38, 9.60; IR (drifts) 3350 (s), 012363 -16- 3293 (m), 2972 (s), 1697 (vs); MS (ES+) m/z (rel. intensity) 358 (M+H+CH3CN*, 55),317 (M+H+, 7), 300 (100); Anal. Calcd for C1SH19N2OZF3: C, 56.96; H, 6.06; N, 8.86.Found: C, 56.86; H, 6.28; N, 8.82.

Example 6 (-) (2R, 4S)-4-Amino-2-ethvl-6-trifluoromethvl-3.4-dihvdro-2H-quinoline-1-carboxvlic acid ethyl ester hemi-(-)-dibenzovl-L-tartrate sait A one iiter flask under nitrogen atmosphère was charged with cis-4- benzyloxycarbonyIamino-2-ethyl-6-trifiuoromethyl-3,4-dihydro-2H-quinoline-1-carboxylic acid ethyl ester (24.0 g, 75.9 mmol, 1.0 equiv) and (-) dibenzoyl-L-tartaricacid (anhydrous) (27.19 g, 75.9 mmol, 1.0 equiv). 300 mL of approximately 97%éthanol (prepared by addirg 10.5 mL H2O to 500 mL absolute éthanol, mixing, andmeasuring out 300 mL) was added. The mixture was stirred at room température for18 h, then filtered. The solids were washed 1 x 48 mL approximately 97% éthanol,and dried to give (-) (2R, 4S)-4-amino-2-ethyl-6-trifluoromethyl-3,4-dihydro-2H-quinoline-1-carboxylic acid ethyl ester hemi-(-)-dibenzoyl-L~tartrate sait as a whitecrystalline solid (14.77 g, 39%); mp 189.5-191.5 °C (dec); 1H NMR (DMSO-d6, 400MHz) δ 0.62 (t, 3H, J=7.3 Hz), 1.16 (t, 3H, J=7.1Hz), 1.3 (m, 3H), 2.5 (m, 1H), 4.1 (m,4H), 5.63 (s, 1H, methine proton in DBTA), 7.47 (m, 2H, DBTA aromatic H’s), 7.6 (m,3H, DBTA aromatic H’s), 7.68 (s, 1H), 7.95 (m, 2H), 8.2 (br s, NH3+, did notintegrate); 13C NMR (DMSO-d6, 100 MHz) δ 169.85,165.53, 154.10,140.14, 134.59,133.51,130.74, 129.69,128.98,126.74,124.82 (q, J=31.7 Hz), 124.69 (q, J=271Hz), 124.50, 120.90, 74.49, 62,14, 53.51, 45.94, 38.81, 28.23, 14.63, 9.58; DEPTspectrum: quaternary carbons δ 169.85,165.53,154.10,140.14,134.59, 130.74,124.82,124.69; CH carbons δ 133.51,129.69, 128.98,126.74,124.50,120.90, 74.49, 53.51,45.94; CH2 carbons δ 62.14, 38.81, 28.23; CH3 carbons δ 14.63, 9.58; IR (drifts) 3278 (m), 2400-3100 (broad), 1703 (vs); MS (ES+) m/z (rel. intensity) 358(M+H+CH3CN+, 55), 317 (M+H+, 7), 300 (100); Anal. Calcd for C15H19N2O2F3.C9H7O4:C, 58.18; H, 5.29; N, 5.65. Found: C, 57.99; H, 5.15; N, 5.64; Chiral HPLC: mobilephase 950:50:2 n-hexane:2-propanol:HOAc, flow rate 1.50 mL/min, column temp40°C, chiralpak™ AD 4.6 x 250 mm, sample concentration approximately 0.5 mg/mLin approximately 1:1 n-hexane:2-propanol. Authentic racemate shows rétention timesof 7.5 min and 10.0 min. (-) (2R, 4S)-4-Amino-2-ethyl-6-trifluoromethyl-3,4-dihydro-2H-quinoline-1-carboxylic acid ethyl ester hemi-(-)-dibenzoyl-L-tartrate sait: 10.0 min, , 072363 -17- 88.9%, 7.5 min «1%, 2.0 min (solvent front) 11.1%; [ct]D = -153 (c=1.07, CH3OH).Example 7 (-)-(2R, 4S)-4-(3,5-Bis-trifluoromethvl-benzylamino)-2-ethvl-6-trifluoromethvl-3,4- dihvdro-2H-quinoline-1-carboxylic acid ethyl ester tosylate sait (-) (2R, 4S)-4-Amino-2-ethyl-6-trifluoromethyl-3l4-dihydro-2H-quinoiine-1 -carboxylicacid ethyl ester hemi-(-)-dibenzoyl-L-tartrate sait (13.0 g, 26.2 mmol, 1.0 equiv) wassuspended in 1,2-dichloroethane (260 mL) in a 500 mL separatory funnel. Themixture was washed 1 x 65 mL 1N NaOH, 1 x 65 mL brine, and dried (MgSO4). Themixture was filtered, concentrated to approximately approximately 80 mL, andtransferred to a 250 mL three neck flask. 3,5-Bis(trifluoromethyl)benzaldehyde (4.53mL, 27.5 mmol, 1.05 equiv) was added, and the mixture stirred 1 h at roomtempérature under nitrogen atmosphère. Sodium triacetoxyborôhydride (11.1 g, 52.4mmol, 2.0 equiv) was added in one portion, and the white slurry was stirred 18 h. 50mL 1,2-dichloroethane and 50 mL 2N NaOH were added, and the aqueous layerextracted 2 x 50 mL 1,2-dichloroethane. The combined organic extracts werewashed 1 x 31 mL 1N HCl, 1 x 50 mL saturated aq. NaHCO3,1 x 50 mL brine, anddried (Na2SO4). The mixture was filtered and concentrated to a clear oil. The oïl wasdissolved in methanol (71 mL). p-Toluenesulfonic acid monohydrate (5.23 g, 27.5mmol, 1.05 equiv) was added. After 5 min, 284 mL isopropyl ether was added. Thesolution was concentrtated to approximately 35mL, transferred to a 500 mL threeneck flask (mech. stirrer), and diluted with 284 mL isopropyl ether. A thick whiteslurry formed in 10 minutes. After stirring 3 h, the slurry was filtered and the cakewashed 2 x 70 mL isopropyl ether. After drying, (-)-(2R, 4S)-4-(3,5-bis-trifluoromethyl-benzylamino)-2-ethyl-6-trifluoromethyl-3,4-dihydro-2H-quino)ine-1-carboxylic acid ethyl ester tosylate sait was isolated as a white powder (16.18 g, 86%overall): mp 191-192'C; 1H NMR (DMSO-d6,400 MHz) δ 0.78 (t, 3H, J=7.5 Hz), 1.21(t, 3H, J=7.0 Hz), 1.5 (m, 3H), 2.24 (s, 3H), 3.08 (m, 1H), 4.17 (m, 2H), 4.41 (m, 1H),4.50 (m, 2H), 4.79 (m, 1H), 7.04 (d, 2H, J=7.9 Hz), 7.42 (d, 2H, J=7.9 Hz), 7.7 (m, 2H), 7.81 (s, 1H), 8.21 (s, 1H), 8.35 (s, 2H), 9.58 (brs, 1H), 9.83 (br s, 1H); 13C NMR(DMSO-d6, 100 MHz) δ 154.00, 145.46, 140.21,138.39, 135.33, 132.51, 131.62,130.79 (q, J=33.2 Hz), 128.49,127.40, 125.82, 125.36, 124.99 (q, J=31.7 Hz), 124.59 (q, J=271 Hz), 123.69 (q, J=273 Hz), 123.44,120.33, 62.32, 53.99, 53.79, 47.98, 33.30, 28.61, 21.13, 14.63, 9.58; DEPT spectrum: quaternary carbons δ 154.00, 145.46,140.21,138.39, 135.33, 130.79, 124.99, 124.59, 123.69; CH carbons « 0123Q3 2. -18- δ 132.51, 131.62,128.49,127.40, 125.82, 125.36,123.44,120.33, 53.99, 53.79; CH2carbons δ 62.32, 47.98, 33.30, 28.61; CH3 carbons δ 21.13,14.63, 9.58; IR (drifts)2300-3100 (broad), 2974 (m), 2731 (m), 2620 (m), 2455 (m), 1714 (s), 1621 (m), 1283 (vs), 1169 (vs), 1126 (vs); MS (ES+) m/z (rel. intensity) 584 (M+H+CH3CN*, 100), 543 (M+H+, 80); Anal. Calcd for C24H23N2O2F9.C7H8O3S·. C, 52.11 ; H, 4.37; N,3.92. Found: C, 52.15; H, 4.22; N, 3.69; [a]D =-77.9 (c= 1.05, CH3OH).

Example 8 (-H2R, 4S)-4-f(3,5-Bis-trifluoromethvl-benzv0-methoxvcarbonvl-amino1-2-ethyl-6- trifluoromethvl-3,4-dihydro-2H-quinoline-1-carboxvlic acid ethvl ester mono ethanolate

Na2CO3 (s) (6.75 g, 63.7 mmol, 3.5 equiv) was added to a room température solutionof (-)-(2R, 4S)-4-(3,5-bis-trifluoromethyl-benzylamino)-2-ethyl-6-trifluoromethyl-3,4-dihydro-2H-quinoline-1-carboxylic acid ethyl ester tosylate sait (13.0 g, 18.2 mmol, 1.0 equiv) in dry THF (130 mL), Methyl chloroformate (3.51 ml_, 45.5 mmol, 2.5equiv) was added neat, dropwise over 2 min. After 24 h, the mixture wasconcentrated to 65 mL, diluted with 260 mL ethyl acetate, and transferred to aseparatory funnel. The mixture was washed 1 x 90 mL 1N HCl (CO2 évolution), 1 x90 mL saturated aq. NaHCO3, 1 x 90 mL brine, and dried (MgSO4). Filtration andconcentration of filtrate afforded a clear oil, which was costripped 3 x 33 mL 2Béthanol. The oil was dissolved in 33 mL 2B éthanol and seeded with a few miiligramsof (-)-(2R, 4S)-4-[(3,5-bis-trifluoromethyI-benzyl)-methoxycarbonyl-amino]-2-ethyl-6-trifluoromethyl-3,4-dihydro-2H-quinoline-1-carboxyIic acid ethyl ester monoethanolate. After stirring 18 h at room température, the slurry was filtered and driedto give (-)-(2R, 4S)-4-[(3,5-bis-trifluoromethyl-benzyl)-methoxycarbonyl-amino]-2-ethyl-6-trifluoromethyl-3,4-dihydro-2H-quinoline-1-carboxylic acid ethyl ester monoethanolate as a white crystalline powder (8.66 g, 74%): mp 54-58 °C; 1H NMR(CDCI3, 400 MHz, 55°C) δ 0.73 (t, 3H, J=7.0 Hz), 1.20 (t, EtOH), 1.27 (t, 3H, J=7.1Hz), 1.42 (m, 2H), 1.66 (m, 1H), 2.25 (br s, 1H), 3.67 (q, EtOH), 3.79 (s, 3H), 4.2 (m,3H), 4.33 (m, 1H), 5.2 (br s, 2H), 7.12 (s, 1H), 7.49 (d, 1H, J=8.3 Hz), 7.57 (d, 1H,J=8.5 Hz), 7.73 (s, 2H), 7.78 (s, 1 H); 13C NMR (CDCI3, 400 MHz) δ 157.74,154.37,141.73,140.05,133.83,132.14 (q, J=33 Hz), 126.94,124.49, 123.96 (q, J=273 Hz),123.13 (q, J=273 Hz), 121.31, 119.17, 62.29, 58.28, 54.42, 53.71, 53.08, 46.67, 37.01, 29.02,18.29,14.32, 9.22, (note: the fourth quartet appears to be buried under the δ 126.94 peak, with J approximately 32 Hz); DEPT spectrum: quaternary carbons -té* * θ 12363 -19- δ 157.74,154.37,141.73,140.05,133.83,132.14,123.96,123.13; CH carbonsδ126.94, 124.49, 121.31, 119.17, 54.42, 53.08; CH2carbons δ 62.29, 58.28,46.67,37.01,29.02; CH3 carbons δ 53.71, 18.29, 14.32, 9.22; IR (drifts) 3489 (s), 2974 (s),2884 (m), 1701 (vs), 1280 (vs), 1131 (vs); MS (ES+) m/z (rel. intensity) 601 (M+H+,100); Anal. Calcd for C26H25N2O4F9.C2H6O: C, 52.01; H, 4.83; N, 4.33. Found: C,51.84; H, 4.54; N, 4.33; chiral HPLC: mobile phase 950:50:2 n-hexane:2-propanol:HOAc, flow rate 1.0 mL/min, 254 nm, chiralpak AD 4.6 x 250 mm, columntemp 40°C, sample concentration approximately 0.5 mg/mL in 90:10 n-hexane:2-propanol, authentic racemate rétention times 3.6 and 4.6 min. (-)-(2R, 4S)-4-[(3,5-Bis-trifluoromethyl-benzyl)-methoxycarbonyl-amino]-2-ethyl-6-trifluoromethyl-3,4-dihydro-2H-quinoline-1-carboxylic acid ethyl ester mono ethanolate shows 4.6 min, 99.1%and 3.6 min, not detected; [a]D = -93.3 (c = 1.08, CH3OH).

Example 9

Anhydrous, (-)-(2R,4S)-4-f(3,5-Bis-trifluoromethyl-benzyl)-methoxycarbonyl-amino1-2- ethvl-6-trifluoromethyl-3.4-dihvdro-2H-quinoline-1-carboxylic acid ethyl ester. A 2.6g portion of 4(S)~[(3,5-bis-trifluoromethyl-benzyl)-methoxycarbonyl-amino]-2(R)-ethyl-6-trifluoromethy[-3,4-dihydro-2H-quinoline-1-carboxy!ic acid ethyl ester (amixture of predominantly amorphous material with traces of ethanolate crystallineform; the title compound was also prepared in an analogous manner starting frompure amorphous or pure ethanolate material) was charged to 13 milliliters of hexanesand heated to effect a solution at about 60°C. The heat was removed and thereaction was allowed to cool to ambient over a one hour period. The reaction wasseeded with anhydrous (-)-(2R,4S)-4-[(3,5-bis-trif)uoromethyl-benzyl)- methoxycarbonyl-amino]-2-ethyl-6-trifluoromethyl-3,4-dihydro-2H-quinoline-1-carboxylic acid ethyl ester and granulated for eighteen hours under ambientconditions. Alternately, the anhydrous crystals may be prepared from hexaneswithout seeding. The product was collected by filtration and air dried. The isolatedproduct X-ray pattern matched the calculated powder pattern.

Density: 1.406

Crystal System: Trigonal

Microscopy: Well formed rods and equant (fractured rods) crystals demonstratinghigh biréfringence when viewed across the C axis. Being in the Trigonal crystal * 012363 -20- system the crystals do not demonstrate biréfringence when viewed down the C axis.The crystals demonstrate a cieavage plane perpendicular to the C axis.

Fusion Microsocopy: In Type A oil------dissolution at 50°C.

Dry---------clear melt at 86°C. NMR: No trace of ethanolateDegree of crystaliinity: Highly crystalline

Hygroscopicity: Non-hygroscopic at 100% relative humidity over48 hours.Appearance: Free flowing white powder.

Example 10

Monoethanolate, (-)-(2R.4S)-4-f(3.5-Bis-trifluoromethyl-benzyl)-methoxycarbonvl- arnino1-2-ethvl-6-trifluoromethyl-3,4-dihvdro-2H-quinoline-1 -carboxylic acid ethvl ester. 4.0 grams of (-)-(2R,4S)-4-[(3,5-bis-trifluoromethyl-benzyl)-methoxycarbonyl-amino]- 2-ethyl-6-trifluoromethyl-3,4-dihydro-2H-quinoline-1-carboxylic acid ethyl ester_weredissolved in 3.5 ml éthanol and sonicated for two minutes to complété dissolution. Awhite soiid formed to which 10 ml éthanol was added and stirred at ambienttempérature overnight. A white powder was filtered and collected on 0.22 pm LSfilter paper followed by washing with about 15 ml. éthanol. The isolated product X-raypattern matched the calculated powder pattern.

Density: 1.402

Crystal System: orthorhombic

Microscopy: crystalline needles with moderate biréfringence.

Fusion Microsocopy: In Type A oil—melt and dissolution at 43°C with loss of water

Dry------------clear melt at 43°C NMR: shows éthanol of solvationDegree of crystaliinity: highly crystallineHygroscopicity: non-hygroscopicAppearance: free-flowing white power

Example 11

Anhydrous (-)-(2R.4ST4-ff3,5-bis-trifluromethvlbenzvl)-methoxvcarbonyl-amino1-2- ethyl-6-trifluoromethvl-3.4-dihvdro-2H-quinoline-1-carboxylic acid ethvl ester. A crude solution of approximately 42 g of (-)-(2R,4S)-4-[(3,5-bis-trifluromethylbenzyl)-methoxycarbonyI-amino]-2-ethyl-6-trifluoromethyl-3,4-dihydro-2H-quinoIine-1- 0/2363 -21- carboxylic acid ethyl ester in 500 ml of ethyl acetate (obtained via the processdescribed in Example 8) was concentrated under vacuum to a volume of 100-135 ml.The remaining ethyl acetate was displaced with 3 X 220 ml 2B EtOH to a final volumeof 100-135 ml. This solution was seeded with a crystal of anhydrous (-)-(2R,4S)-4-[(3,5-bis-trifIuromethylbenzyl)-methoxycarbonyI-amino]-2-ethyl-6-trifluoromethyl-3,4-dihydro-2H-quinoline-1 -carboxylic acid ethyl ester. After stirring 18 hr at roomtempérature the slurry was filtered and vacuum dried to give 19.81 g of anhydrous (-)-(2R,4S)-4-[(3,5-bis-trifluromethylbenzyl)-methoxycarbonyl-amino]-2-ethyl-6-trifluoromethyl-3,4-dihydro-2H-quinoline-1 -carboxylic acid ethyl ester. The meitingpoint behaviour was the same as the matériel prepared via Example 9 confirming theanhydrous nature of the material.

Atorvastatin or its cyclized lactone form may readily be prepared as describedin U.S. Patent No. 4,681,892, which is incorporated herein by référencé, Thehemicalcium sait of atorvastatin, which is currently sold as Lipitor®, may readily beprepared as described in U.S. Patent No. 5,273,995, which is incorporated herein byréférencé.

The hydroxylated dérivatives (métabolites) of atorvastatin (or the cyclizedlactone form or pharmaceutically acceptable salts thereof) may be prepared asdescribed in U.S. pat. No. 5,385,929. The ortho, meta and para hydroxy dérivativesare encompassed herein: (2R-trans)-5-(4-fluorophenyl)-2-(1-methyiethyl)-N-(2-hydroxyphenyi)-4-phenyl- 1-[2-(tetrahydro-4-hydroxy-6-oxo-2H-pyran-2-yl)ethyl}-1H-pyrroie-3-carboxamide. (2R-trans)-5-(4-fluorophenyl)-2-(1-methylethyl)-N-(3-hydroxyphenyl)-4-phenyl-1 -[2-(tetrahydro-4-hydroxy-6-oxo-2H-pyran-2-yl)ethyl}-1 H-pyrrole-3-carboxamide; and (2R-trans)-5-(4-fluorophenyl)-2-(1-methylethyl)-N-(4-hydroxyphenyl)-4-phenyl-1 -[2-(tetrahydro-4-hydroxy-6-oxo-2H-pyran-2-yl)ethyl]-1 H-pyrrole-3-carboxam ide.

The expression “pharmaceutically acceptable salts” inciudes bothpharmaceutically acceptable acid addition salts and pharmaceutically acceptablecationic salts. The expression "pharmaceutically-acceptable cationic salts" isintended to define but is not limited to such salts as the alkali métal salts, (e.g.sodium and potassium), alkaline earth métal salts (e.g. calcium and magnésium),aluminum salts, ammonium salts, and salts with organic amines such as benzathine(Ν,Ν-dibenzylethylenediamine), choline, diethanolamine, ethylenediamine,meglumine (N-methyiglucamine), benethamine (N-benzyiphenethylamine), • ÛÎ2363 -22- diethylamine, piperazine, tromethamine (2-amino-2-hydroxymethyl-1,3-propanediol)and procaine. The expression "pharmaceutically-acceptable acid addition salts" isintended to define but is not limited to such salts as the hydrochloride, hydrobromide,sulfate, hydrogen sulfate, phosphate, hydrogen phosphate, dihydrogenphosphate,acetate, succinate, citrate, methanesulfonate (mesylate) and p-toluenesulfonate(tosylate) salts.

Other pharmaceutically-acceptable cationic salts of atorvastatin may bereadily prepared by reacting the free acid form of atorvastatin with an appropriatebase, usually one équivalent, in a co-solvent. Typicai bases are sodium hydroxide,sodium methoxide, sodium ethoxide, sodium hydride, potassium methoxide,magnésium hydroxide, calcium hydroxide, benzathine, choline, diethanolamine,piperazine and tromethamine. The sait is isolated by concentration to dryness or byaddition of a non-solvent. In many cases, salts are preferably prepared by mixing asolution of the acid with a solution of a different sait of the cation (e.g., sodium orpotassium ethylhexanoate, magnésium oleate) and employing a solvent (e.g., ethylacetate) from which the desired cationic sait précipitâtes. The salts may afso beisolated by concentrating the reaction solution and/or by adding a non-solvent.

The acid addition salts of atorvastatin may be readily prepared by reacting thefree base form of atorvastatin with the appropriate acid. When the sait is of amonobasic acid (e.g., the hydrochloride, the hydrobromide, the p-toluenesulfonate,the acetate), the hydrogen form of a dibasic acid (e.g., the hydrogen sulfate, thesuccinate) or the dihydrogen form of a tribasic acid (e.g., the dihydrogen phosphate,the citrate), at least one moiar équivalent and usually a molar excess of the acid isempioyed. However when such salts as the sulfate, the hemisuccinate, the hydrogenphosphate or the phosphate are desired, the appropriate and exact Chemicaléquivalents of acid will generally be used. The free base and the acid are usuallycombined in a co-solvent from which the desired sait précipitâtes, or can be otherwiseisolated by concentration and/or addition of a non-solvent.

In addition, [2R, 4SJ4-[(3,5-bis-trifluoromethyl-benzyl)-methoxycarbonyl- amino]-2-ethyl-6-trifluoromethyI-3,4-dihydro-2H-quinoiine-1-carboxylic acid ethyl ester can exist as a monoethanolate and an anhydrous form as described in provisional U.S. application serial no. 60/167,967 and such forms are within the scope of the invention. & 0123$$ -23-

Atorvastatin, or the cyclized lactone form the ortho, meta and para hydroxydérivatives of said compounds and pharmaceuticaliy acceptable salts thereof mayoccur as hydrates or solvatés. Said hydrates and solvatés are also within the scopeof the invention.

The pharmaceutical combinations and methods of this invention are ailadapted to therapeutic use as agents in the treatment of atherosclerosis, anginapectoris, and a condition characterized by the presence of both low HDL levels andhyperlipidemia in mammals, particularly humans. Further, since these diseases andconditions are closely related to the development of cardiac disease and adversecardiac conditions, these combinations and methods, by virtue of their action asantiatherosclerotics, antianginals and antihyperlipidemics, are useful in themanagement of cardiac risk as well as mixed lipid disorders, such as those seen indiabètes and other metabolic syndromes.

The utility of the compounds of the présent invention as medical agents in thetreatment of atherosclerosis in mammals (e.g. humans) is demonstrated by theactivity of the compounds of this invention in conventional assays and the clinicalprotocol described below:

In the following protocols référencé to a CETP inhibitor X is to [2R, 4S]4-[(3,5-bis-trifluoromethyl-benzyl)-methoxycarbonyl-amino3-2-ethyl-6'trifluoromethyl-3,4-dihydro-2H-quinoline-1-carboxylic acid ethyl ester.

Atherosclerosis Protocol

This study is a prospective randomized évaluation of the effect of acombination of CETP inhibitor X and atorvastatin (or its metabolities) on theprogression/regression of atherosclerotic disease. The study is used to show that acombination of CETP inhibitor X and atorvastatin (or its metabolities) are effective inslowing or arresting the progression or causing régression of existing atheroscleroticdisease as evidenced by changes in plaque and/or lumen parameters via variousimaging techniques, coronary angiography or carotid ultrasound, in subjects with orwithout established disease.

This study is an imaging documentation of atherosclerotic disease carried out as a doubie-blind trial of a minimum of about 500 subjects and preferably of about 780 to about 1200 subjects. lt is especially preferred to study about 1200 subjects in this study. Subjects are admitted into the study after satisfyïng certain entry criteria set forth below. .012363 -24-

Entry criteria: Subjects accepted for entry into this trial must satisfy certaincriteria. Thus the subject must be an adult, either male or female, aged 18-80 yearsof âge in whom cardiovascular imaging is clinically indicated. Subjects will hâveevidence of atherosclerotic disease that is judged not likely to require intervention 5 over the next 3 years. It is required that the vessels undergoing analysis hâve notbeen interfered with. Since percutaneous transluminal cardiac angioplasty (PTCA)interfères with segments by the insertion of a balloon cathéter, non-PTCA segmentsare required for analysis. It is also required that the vessels to be analyzed hâve notsuffered a thrombotic event, such as a myocardial infarct (Ml). Thus the requirement 10 for non-MI vessels. Potential areas to be analyzed include: left main, proximal, midand distal left anterior descending, first and second diagonal branch, proximal anddistal left circumflex, first or largest space obtuse marginal, proximal, mid and distalright coronary artery.

Generally, due to the number of patients and the physical limitations of any 15 one facility, the study is carried out at multiple sites. At entry into the study, subjects undergo quantitative coronary as well as carotid artery and/or peripheral vesselimaging at designated testing centers. This establishes baselines for each subject.Once admitted into the test, subjects are randomized to receive either CETP inhibitorX (10 -100 mgs) with atorvastatin calcium (10 - 80 mgs) or its métabolites (.02 20 mg/kg-200 mg/kg), each one separately, and/or neither. Ail doses set forth in this protocol are per day doses. The amount of CETP inhibitor X or atorvastatin (or itsmetaboiites) may be varied as required.

The subjects are monitored for a one to three year period, generally threeyears being preferred. Imaging assessment of vessels that does not require an 25 invasive procedure are performed at regular intervals throughout the study.

Generally, six month intervals are suitable. Typically this assessment is performed using B-mode ultrasound and/or équivalent equipment However, aperson skilled in the art may use other methods of performing this assessment.Invasive imaging is performed at the conclusion of the one to three year treatment 30 period. The baseline and post-treatment images are evaluated for new lésions or progression of existing atherosclerotic lésions.

The primary objective of this study is to show that the combination of CETP inhibitor X and atorvastatin (or metaboiites thereof) or pharmaceutically acceptable salts thereof reduces the progression of atherosclerotic lésions as measured by 012363 -25- quantitative coronary angiography (QCA) or CBCT, or 1WS in subjects with ciinicalcoronary artery disease. These techniques measure the amount of atherosclerosis ina vessei.

The primary endpoint of the study is the change in atherosclerotic burden ofthe affected vessei. Using QCA as an example, the diameter of an arterial segmentis measured at various portions along the length of that segment. The averagediameter of that segment is then determined. After the average segment diameter ofmany segments has been determined, the average of ail segment averages isdetermined to arrive at the average mean segment diameter. The mean segmentdiameter of subjects taking atorvastatin (or its métabolites) or pharmaceuticallyacceptable salts thereof and the CETP inhibitor X will décliné more slowiy, will behalted completely, or there will be an increase in the mean segment diameter. Theseresults represent slowed progression of atherosclerosis, halted progression ofatherosclerosis and régression of atherosclerosis, respectively.

The secondary objective of this study is to show that the combination of theCETP inhibitor X and atorvastatin (or its métabolites) or a pharmaceutically saitthereof reduces the rate of progression of atherosclerosis in other arteries. Forexample, using carotid arteries as an example, as measured by the slope of themaximum intimal-medial thickness measurements averaged over 12 separate wallsegments (Mean Max) as a function of time, more than does the CETP inhibitor X oratorvastatin (or its métabolites) or pharmaceutically acceptable sait thereof, alone.The intimal-medial thickness of subjects taking atorvastatin (or its métabolites) orpharmaceutically acceptable sait thereof and the CETP inhibitor X will increase moreslowiy, will cease to increase or will decrease. These results represent slowedprogression of atherosclerosis, halted progression of atherosclerosis and régressionof atherosclerosis, respectively.

The utility of the compounds of the présent invention as medical agents in thetreatment of angina pectoris in mammals (e.g., humans) is demonstrated by theactivrty of the compounds of this invention in conventional assays and the ciinicalprotocol described below:

Angina Protocol

This study is a double blind, parallel arm, randomized study to show theeffectiveness of the CETP inhibitor X and atorvastatin (or its métabolites) or 012363 * -26- pharmaceutically acceptable salts thereof given in combination in the treatment ofsymptomatic angina.

Entrv criteria: Subjects are males or females between 18 and 80 years of âgewith a history of typical chest pain associated with one of the following objectiveévidences of cardiac ischemia: (1) stress test segment élévation of about onemillimeter or more from the ECG; (2) positive treadmili stress test; (3) newwallmotion abnormality on ultrasound; or (4) coronary angiogram with a significantqualifying stenosis. Generally a stenosis of about 30-50% is considered to besignificant.

Each subject is evaluated for about ten to thirty-two weeks. At least tenweeks are generally required to complété the study. Sufficient subjects are used inthis screen to ensure that about 200 to 800 subjects and preferably about 400 subjectare evaluated to complété the study. Subjects are screened for compliance with theentry criteria, set forth below, during a four week run in phase. After the screeningcriteria are met, subjects are washed out from their current anti-anginal médicationand stabilized on a long acting nitrate such as, for example, nitroglycerin, isosorbide-5-mononitrate or isosorbide dinitrate. The term “washed out”, when used inconnection with this screen, means the withdrawal of current anti-anginal médicationso that substantially ail of said médication is eliminated from the body of the subject. A period of eight weeks is preferably allowed for both the wash out period and for theestablishment of the subject on stable doses of said nitrate. Subjects having one ortwo attacks of angina per week while on stable doses of long acting nitrate aregenerally permitted to skip the wash out phase. After subjects are stabilized onnitrates, the subjects enter the randomization phase provided the subjects continue tohâve either one or two angina attacks per week. In the randomization phase, thesubjects are randomly placed into one of the four arms of the study set forth below.After completing the wash out phase, subjects in compliance with the entry criteriaundergo twenty four hour ambulatory electrocardigram (ECG) such as Holtermonitoring, exercise stress testing such as a treadmili and évaluation of myocardialperfusion using PET (photon émission tomography) scanning to establish a baselinefor each subject. When conducting a stress test, the speed of the treadmili and thegradient of the treadmili can be controlled by a technician. The speed of the treadmiliand the angle of the gradient are generally increased during the test. The time 012363 -27-

Wo υζ, intervals between each speed and gradient increase is generally determined using amodified Bruce Protocol.

After the baseline investigations hâve been compieted, subjects are initiatedon one of the following four arms of the study: (1) placebo; (2) atorvastatin calcium 5 (about 2.5 mg to about 160 mg) or its métabolites (.02 mg/kg-200 mg/kg) ; (3) CETPinhibitor X (about 10 mg to about 120 mg); or (4) a combination of the above doses ofCETP inhibitor X and atorvastatin calcium (or its métabolites) together. The subjectsare then monitored for two to twenty-four weeks.

After the monitoring period has ended, subjects will undergo the following 10 investigations: (1) twenty four hour ambulatory ECG, such as Holter monitoring; (2)exercise stress testing (e.g. treadmill using said modified Bruce Protocol); and (3)évaluation of myocardial perfusion using PET scanning. Patients keep a diary ofpainful ischémie events and nitroglycérine consumption. It is generally désirable tohâve an accurate record of the number of anginal attacks suffered by the patient 15 during the duration of the test. Since a patient generally takes nitroglycerin to ease the pain of an anginal attack, the number of times that the patient administersnitroglycérine provides a reasonably accurate record of the number of anginalattacks.

To demonstrate the effectiveness of the compound combination of this 20 invention, and to détermine the dosage amounts of the compound combination of thisinvention, the person conducting the test will evaluate the subject using the testsdescribed. Successful treatment will yield fewer instances of ischémie events asdetected by ECG, will allow the subject to exercise longer or at a higher intensity levelon the treadmill, or to exercise without pain on the treadmill, or will yield better 25 perfusion or fewer perfusion defects on photoemission tomography (PET).

The utility of the compounds of the présent invention as medical agents in the treatment of lipid abnormalities in mammals (e.g., humans) suffering from acombination of low HDL-C and high LDL-C is demonstrated by the activity of thecompounds of this invention in conventional assays and the clinical protocol 30 described below:

Dvslipidemia Protocol

This study is a double blind, paraliel arm, randomized study to show the effectiveness of CETP inhibitor X and atorvastatin calcium (or its métabolites) or pharmaceutically acceptable salis thereof given in combination in controlling both low 012363 -28-

'S V HDL-C and high LDL-C in subjects who hâve mild, moderate, or severe levels ofthese iipid abnoimalities.

Each subject is evaluated for 10 to 20 weeks and preferabiy for 14 weeks.Sufficient subjects are used in this screen to ensure that about 400 to 800 subjects 5 are evaluated to complété the study.

Entrv criteria: Subjects are male or female adults between 18 and 80 years of âge having both low HDL-C and high LDL-C. The presence of these abnormalities isevidenced by évaluation of the low density lipoprotein (LDL) level of the subjectrelative to certain positive risk factors and évaluation of their HDL-C levels.. If the 10 subject has no coronary heart disease (CHD) and has less than two positive riskfactors, then the subject is considered to hâve high LDL if the LDL of the subject isgreater than or equal to 190 mg/dl. If the subject has no CHD and has two or morepositive risk factors, then the subject is considered to hâve hyperlipidemia if the LDLof the subject is greater than or equal to 160 mg/dl. If the subject has CHD, then the 15 subject is considered to hâve hyperlipidemia if the LDL of the subject is greater than or equal to 130.

Positive risk factors include (1) male over 45, (2) female over 55 wherein saidfemale is not undergoing hormone replacement therapy (HRT), (3) family history ofprématuré cardiovascular disease, (4) the subject is a current smoker, (5) the subject 20 has diabètes, (6) an HDL of less than 35, and (7) the subject has hypertension. AnHDL of greater than 60 is considered a négative risk factor and will offset one of theabove mentioned positive risk factors.

The presence of low HDL is evidenced by a level less than 35 mg/dl.

Subjects are screened for compliance with the entry criteria set forth above. 25 After ail screening criteria are met, subjects are washed out from their current Iipidlowering médication and are placed on the NCEP ATP II Step 1 diet. The NCEP ATPIl (adult treatment panel, 2nd révision) Step 1 diet sets forth the amount of saturatedand unsaturated fat which can be consumed as a proportion of the total calorieintake. The term “washed out" where used in connection with this screen, means the 30 withdrawal of current Iipid lowering médication so that substantially ail of said médication is eliminated from the body of the subject. Newiy diagnosed subjects generally remain untreated until the test begins. These subjects are also placed on the NCEP Step 1 diet. After the four week wash out and diet stabilization period, subjects undergo the following baseline investigations: (1) medical history and (2) 012363 -29- fasting lipid screen. The fasting lipid screen détermines baseline lipid levels in thefasting State of a subject. Generally, the subject abstains from food for twelve hours,at which time lipid levels are measured.

After the baseline investigations are performed subjects are started on one ofthe foliowing: (1) a fixed dose of CETP inhibitor X, generally about 10 to 120 mg; (2)a fixed dose of atorvastatin calcium, generally about 10 to 80 mg or its métabolites(.02 mg/kg-200 mg/kg); or (3) a combination of the above doses of CETP inhibitor Xand atorvastatin calcium (or its métabolites) together. Subjects remain on thesedoses for a minimum of six weeks, and generally for no more than eight weeks. Thesubjects return to the testîng cénter at the conclusion of the six to eight weeks so thatthe baseline évaluations can be repeated. The lipid screen measures the totalcholestérol, LDL-cholesterol, HDL-cholesterol, triglycérides, apoB, VLDL (very lowdensity lipoprotein) and other components of the lipid profile of the subjectImprovements in the values obtained after treatment relative to pretreatment valuesindicate the utility of the compound combination.

The utility of the compounds of the présent invention as medical agents in themanagement of cardiac risk in mammals (e.g., humans) at risk for an adversecardiac event is demonstrated by the activity of the compounds of this invention inconventional assays and the clinical protocol described below:

Risk of Future Cardiovascular Events

This study is a double blind, parallel arm, randomized study to show theeffectiveness of the CETP inhibitor X and atorvastatin (and its métabolites) orpharmaceutically acceptable salts thereof given in combination in reducing the overallcalculated risk of future events in subjects who are at risk for having futurecardiovascular events. This risk is calculated by using the Framingham RiskEquation. A subject is considered to be at risk of having a future cardiovascularevent if that subject is more than one standard déviation above the mean ascalculated by the Framingham Risk Equation. The study is used to evaluate theefficacy of a fixed combination of the CETP inhibitor X and atorvastatin (or itsmétabolites) in controlling cardiovascular risk by controlling both low HDL and highLDL in patients who hâve both mild to moderate abnormalities in these lipids.

Each subject is evaluated for 10 to 20 weeks and preferably for 14 weeks.

Sufficient subjects are recruited to ensure that about 400 to 800 subjects are evaluated to complété the study. 012363 -30

Entrv criteria: Subjects included in the study are male or female adult subjectsbetween 18 and 80 years of âge with a baseline five year risk which risk is above themédian for said subject's âge and sex, as defined by the Framingham Heart Study,which is an ongoing prospective study of aduit men and women showing that certain 5 risk factors can be used to predict the development of coronary heart disease. Theâge, sex, systolic and diastolic blood pressure, smoking habit, presence or absenceof carbohydrate intolérance, presence or absence of left ventricuiar hypertrophy,sérum cholestérol and high density lipoprotein (HDL) of more than one standarddéviation above the norm for the Framingham Population are ail evaluated in 10 determining whether a patient is at risk for adverse cardiac event. The values for the risk factors are inserted into the Framingham Risk équation and caiculated todétermine whether a subject is at risk for a future cardiovascular event.

Subjects are screened for compliance with the entry criteria set forth above.After ail screening criteria are met, patients are washed out from their current lipid 15 lowering médication and any other médication which wïll impact the résulte of the screen. The patients are then placed on the NCEP ATP II Step 1 diet, as describedabove. Newly diagnosed subjects generally remain untreated until the test begins.These subjects are also placed on the NCEP ATP II Step 1 diet. After the four weekwash out and diet stabilization period, subjects undergo the foliowing baseline 20 investigations: (1) blood pressure; (2) fasting; (3) lipid screen; (4) glucose tolérancetest; (5) ECG; and (6) cardiac ultrasound. These tests are carried out using standardprocedures well known to persons skilled in the art. The ECG and the cardiacultrasound are generally used to measure the presence or absence of left ventricuiarhypertrophy. 25 After the baseline investigations are performed patients will be started on one of the foliowing: (1) a fixed dose of CETP inhibitor X (about 10-120 mg); (2) a fixeddose of atorvastatin (about 10 to 80 mg) or its métabolites (.02 mg/kg-200 mg/kg); or(3) the combination of the above doses of CETP inhibitor X and atorvastatin (or itsmétabolites). Patients are kept on these doses and are asked to return in six to eight 30 weeks so that the baseline évaluations can be repeated. At this time the new values are entered into the Framingham Risk équation to détermine whether the subject has a lower, greater or no change in the risk of future cardiovascular event. The above assays demonstrating the effectiveness of [2R, 4SJ4-[(3,5-bis-trifluoromethyl-benzyl)- methoxycarbonyI-amino]-2-ethyl-6-trifluoromethyl-3.4-dihydro-2H-quinoline-1- 012363 -31- carboxylic acid ethyl ester and atorvastatin or hydroxy dérivatives thereof orpharmaceutically acceptable saits thereof in the prévention and/or treatment ofangina pectoris, atheroscierosis, low HDL and high LDL together, and themanagement of cardiac risk, also provide a means whereby the activities of thecompounds of this invention can be compared between themselves and with theactivities of other known compounds. The resuits of these comparisons are useful fordetermining dosage levels in mammais, inciuding humans, for the prévention and/ortreatment of such diseases. in general, [2R, 4S]4-[(3,5-bis-trifluoromethyl-benzyl)-methoxycarbonyi-amino]-2-ethyl-6-trifluoromethyl-3,4-dihydro-2H-quinoline-1-carboxylic acid ethyl esteris administered in a dosage in the range of about 0.1 to about 10 mg/kg/daypreferably about 0.5 to about 5 mg/kg/day. in general atorvastatin or the cyclized lactone form or its pharmaceuticallyacceptable salts, is administered in a dosage of about 2.5 mg/day to about 160mg/day. Preferably, atorvastatin calcium is administered in a dosage of about 10mg/day to about 80/mg day. Typicaliy the hydroxy métabolites of these compoundsare administered in a dosage of about .02 mg/kg/day-200 mg/kg/day. These dosagesbeing based on an average human subject having a weight of about 65 to about 70kg.

The compounds of the présent invention are generally administered in theform of a pharmaceutical composition comprising at least one of the compounds ofthis invention together with a pharmaceutically acceptable carrier, vehicle or diluent.Thus, the compounds of this invention can be administered either individually ortogether in any conventional oral, parentéral or transdermal dosage form.

For oral administration a pharmaceutical composition can take the form ofsolutions, suspensions, tablets, pills, capsules, powders, and the like. Tabletscontaining various excipients such as sodium citrate, calcium carbonate and calciumphosphate are employed along with various disintegrants such as sta'rch andpreferably potato or tapioca starch and certain complex silicates, together withbinding agents such as polyvinylpyrrolidone, sucrose, gelatin and acacia.

Additionally, lubricating agents such as magnésium stéarate, sodium lauryl sulfate and talc are often very useful for tabletting purposes. Solid compositions of a similar type are also employed as fillers in soft and hard-filled gelatin capsules; preferred materiais in this connection also include lactose or milk sugar as well as high 012363 -32- molecular weight polyethylene glycols. When aqueous suspensions and/or élixirs aredesired for oral administration, the compounds of this invention can be combined withvarious sweetening agents, flavoring agents, colorïng agents, emulsifying agentsand/or suspending agents, as weil as such diiuents as water, éthanol, propylenegiycol, glycerin and various like combinations thereof.

The combination of this invention may also be administered in a controliedrelease formulation such as a slow release or a fast release formulation. Suchcontrolied release dosage formulations of the combination of this invention may beprepared using methods well known to those skiiled in the art. The method ofpreferred administration will be determined by the attendant physician or other personskiiled in the art after an évaluation of the subject’s condition and requirements. Thegenerally preferred formulation of atorvastatin is Lipitor®. For [2R, 4S]4-[(3,5-bis-trifluoromethyl-benzyi)-metboxycarbonyI-amino]-2-ethyl-6-trifluoromethyl-3.4-dihydro-2H-quinoline-1-carboxylic acid ethyl ester a generally preferred formulation is adosage unit form in a capsule, for example a gel capsule, it may contain, in additionto or instead of materials of the above type, a liquid carrier such as a fatty glycerideor mixtures of fatty glycerides, such as olive oil, or Miglyol TM or Capmul TMglycerides. Dosage forms may also include oral suspensions.

For purposes of parentéral administration, solutions in sesame or peanut oilor in aqueous propylene giycol can be employed, as well as stérile aqueous solutionsof the corresponding water-soluble salts. Such aqueous solutions may be suitablybuffered, if necessary, and the liquid diluent first rendered isotonie with sufRcientsaline or glucose. These aqueous solutions are especially suitable for intravenous,intramuscular, subeutaneous and intraperitoneal injection purposes. in thisconnection, the steriie aqueous media employed are ail readily obtainable bystandard techniques well-known to those skiiled in the art.

Methods of preparing various pharmaceutical compositions with a certainamount of active ingrédient are known, or will be apparent in light of this disclosure, tothose skiiled in this art. For examples, see Reminqton's Pharmaceutical Sciences,Mack Publishing Company, Easter, Pa., 15th Edition (1975).

Pharmaceutical compositions accordtng tothe invention may contain O.1%- 95% of the compound(s) of this invention, preferably 1%-70%. In any event, the composition or formulation to be administered will contain a quantity of a 012363 -33- compound(s) according to the invention in an amount effective to treat the conditionor disease of the subject being treated.

Since the présent invention relates to the treatment of diseases andconditions with a combination of active ingrédients which may be administered 5 separately, the invention aiso relates to combining separate pharmaceutical compositions in kit form. The kit inciudes two separate pharmaceutical compositions:[2R,4S]4-[(3,5-bis-trifluoromethyl-ben2yl)-methoxycarbonyl-aminoî-2-ethyl-6-trifiuoromethyl-3.4-dihydro-2H-quinoline-1-carboxylic acid ethyi ester and atorvastatin(or its metaboiites) or a pharmaceuticaily acceptable sait thereof. The kit inciudes 10 means for containing the separate compositions such as a divided bottle or a dividedfoil packet, however, the separate compositions may also be contained within asingle, undivided container. Typically the kit inciudes directions for the administrationof the separate components. The kit form is particularly advantageous when theseparate components are preferably administered in different dosage forms (e.g., 15 oral and parentéral), are administered at different dosage intervais, or when titrationof the individual components of the combination is desired by the prescribingphysician.

It shouid be understood that the invention is not iimited to the particularembodiments described herein, but that various changes and modifications may be 20 made without departing from the spirit and scope of this novel concept as defined bythe foiiowing daims.

Claims (19)

1. 012363 -34- CLAIMS

   1. A pharmaceutical composition comprising a therapeutically effective amount of acomposition comprising: a. [2R, 4S]4-[(3,5-bis-trifluoromethyl-benzyl)-methoxycarbonyi-amino]5 2-ethyl-6-trifluoromethyl-3,4-dihydro-2H-quinoiine-1-carboxyIic acid ethyl ester; b. a compound of the Formula I

   Formula I 10 or, the open chain Formula IA 012363 -35-

   wherein R1 is hydrogen or hydroxy or the pharmaceutically acceptable salts 5 thereof; and c. a pharmaceutically acceptable carrier, vehicle or diluent.
2. 2. A pharmaceutical composition as recited in claim 1 wherein R1 is hydrogen or apharmaceutically acceptable sait thereof.
3. 3. A pharmaceutical composition as recited in claim 2 comprising the hemicalcium 10 sait of atorvastatin.
4. 4. A pharmaceutical composition as recited in claim 1 wherein R1 is 2-hydroxy or apharmaceutically acceptable sait thereof.
5. 5. Use of 15 a a first compound, said first compound being [2R, 4S]4-[(3,5-bis- trifluoromethyl-benzyl)-methoxycarbonyl-amino]-2-ethyl-6-trifluoromethyl-3,4-dihydro2H-quinoline-1-carboxylic acid ethyl ester; and b. a second compound, said second compound being a compoundhaving the Formula I 012363 -36-

   Formula I or, the open chain Formula IA

   5 ,, .01,2363 -37- wherein R1 is hydrogen or hydroxy or the pharmaceutically acceptable saltsthereof ; and wherein said first compound and said second compound are each optionaliy andindependently administered togetner with a pharmaceutically acceptable carrier,vehicle or diluent in the manufacture of a médicament for treating a mammal in need oftherapeutic treatment.
6. 6. Use as recited in claim 5 wherein R1 is hydrogen or a pharmaceuticallyacceptable sait thereof.
7. 7. Use as recited in claim 6 comprising the hemicalcium sait of atorvastatin.
8. 8. Use as recited in claim 5 wherein R1 is 2-hydroxy or a pharmaceutically acceptable sait thereof.
9. 9. Use as recited in claim 5 wherein atherosclerosis is prevented or treated.
10. 10. Use as recited in claim 5 wherein the progression of atherosclerotic plaques is slowed.
11. 11. Use as recited in claim 10 wherein the treatment of atherosclerosis causesthe régression of atherosclerotic plaques.
12. 12. Use as recited in claim 5 wherein the therapeutic treatment comprises HDLélévation treatment and antihyperlipidemic treatment.
13. 13. Use as recited in claim 5 wherein angina is prevented.
14. 14. Use as recited in claim 5 wherein the therapeutic treatment comprisescardiac risk management.
15. 15. A kit for achieving a therapeutic effect in a mammal comprising a therapeuticaliyeffective amount of a composition comprising: a. [2R, 4S]4-[(3,5-bis-trifiuoromethyi-bsnzyl)-methoxycarbonyl-arnino}-2-ethyl-6-trifiuoromethyi-3,4-dihydro-2H-quinoiine-1-carboxyiic acid ethyl ester and apharmaceutically acceptable carrier, vehicle or diluent in a first unit dosage form; b. a compound having the Formula I 0.12363 -38-

    R' Formula I or, the open chain Formula IA

    5 - 012363 -39- 10 15 wherein R1 is hydrogen or hydroxy or the pharmaceutically acceptable salts thereof and a pharmaceutically acceptable carrier, vehicle or diluent in a second unit dosageform; and c. means for containing said first and second dosage forms.
16. 16. A kit as recited in daim 15 wherein R1 is hydrogen or a pharmaceuticallyacceptable sait thereof.
17. 17. A kit as recited in claim 16 comprising the hemicalcium sait of atorvastatin.
18. 18. A kit as recited in claim 15 wherein R1 is 2-hydroxy or a pharmaceuticallyacceptable sait thereof.
19. 19. A first pharmaceutical composition for use with a second pharmaceuticalcomposition for achieving a therapeutic effect in a mammal, which effect is greaterthan the individual therapeutic effects achieved by administering said first or secondpharmaceutical compositions separately and which second pharmaceuticalcompositions comprises an amount of a Formula I or IA compound or apharmaceutically acceptable sait thereof having the Formula I

    Formula I