CA2077226A1 - N,n'n'-trisubstituted-5-bis- aminomethylene-1,3-dioxane-4, 6-dione inhibitors of acyl-coa: cholesterol-acyl transferase - Google Patents

Abstract

Abstract of the Disclosure A compound of the formula:

in which X, Y and Z are, independently, hydrogen, halogen, hydroxy, nitro, cyano, carboxyl, trifluoromethyl, phenyl, amino, alkylamino, dialkylamino, alkyl or alkoxy; R1 is alkyl, alkenyl, cycloalkyl, phenyl, benzyl or substituted phenyl or benzyl where the substituents are alkyl or alkoxy; R2 is , or

Description

2~.?'~

, N,~ TRISUBSTITUTED-5-~1~-AMINOMETHYLENE-1,3-DIOXANE-4,6-DIONE INHIBITORS OF ACYL-CoA:CHOLE~l'EROL-ACYL TRANSFERASE
S .
Back~rQund of the Invention This invention relates to chemical compounds which display inhibition of Acyl-Coenzyme A: Cholesterol Acyltransferase (ACAT). Compounds of this type aid in 10 reducing cholesterol absorption and its effect on atherosclerosis.

Atherosclerosis is the most common form of arteriosclerosis and is characterized by the buildup of phospholipids and esterified cholesterol in large and medium arteries causing them to be inelastic and thus weakened. These inelastic and 15 occluded arteries are the most common cause of ischemic heart disease.

ACAT is an important enzyme for the intracellular es~erification of cholesterol.Studies of this enzyme in cultured cells (M.S. Brown, J. Biol. Chem. 1980, 255, 9344) has shown that increases in ACAT activity represent increases in the presence of 20 cholesterol laden lipoproteins. Regulation of ACAT helps prevent the absorption of cholesterol in the intestinal mucosa, and assists in the reversal of already present atherosclerotic lesions.

~escription of thç Invention In accordance with this invention, there is provided a group of diarninomethylene dioxane dione derivatives of formula 1:
X

O O

d~R
X ~ R2 Y z Z~37 in which X, Y and Z are, independently, hydrogen, halogen, hydroxy, nitro, cyano, carboxyl, trifluoromethyl, phenyl,~amino, alkylamino of 1 to 12 carbon atoms, diaL~cylamino in which each alkyl group has 1 to 12 carbon atoms, alkyl of 1 to 12 carbon atoms or alkoxy of 1 to 12 carbon atoms;
Rl is hydrogen,alkyl of 1 to 18 carbon atoms, alkenyl of 2 to 18 carbon atoms, cycloalkyl of 5 to 8 carbon atoms, phenyl, benzyl or substituted phenyl or benzyl where the substituents are alkyl of 1 to 12 carbon atoms or alkoxy of 1 to 12 carbon atoms;
R2 is ~NX~X ? I~Z ~Y

Z, X or Z
or a pharmaceutically acceptable salt thereof.

The halogen substituent referred to above may be chlorine, bromine, fluorine or iodine, fluorine being preferred. The pharmaceutically acceptable salts are derived from known inorganic or organic acids such as hydrochloric, hydrobromic, sulfuric, nitric, phosphonc, methanesulfonic, ethanesulfonic, hydroxyethanesulfonic, toluene sulfonic, naphthalenesulfonic, formic, acetic, propionic, oxalic, succinic, glycollic, lactic, malic, tartaric, citric, ascorbic, maleic, hydroxyrnaleic, pyruvic, phenylacetic, benzoic, para-amino benzoic, para-hydroxybenzoic, salicylic, sulfanilic acids, and the like.
Of these compounds, those preferred of the quinolinyl substituted series, on thebasis of their in ~tro and in vivo potency are those of formula 3:

23! ~ ~r~

0~0 0~0 .1 in which X and Z are, independently, alpha branched alkyl of 1 to 6 carbon atoms;
Rl is alkyl of 1 to 18 carbon atoms;
5 and R2 is 2-, 3- or 4-quinolinyl;
or a pharmaceutically acceptable salt thereo~.

The preferred compounds containing the thienyl substituents and those of formula S:

O o O~o X~N N ~R, 15 in which X, Y and Z are, independently, hydrogen, alkyl of 1 to 6 carbon atoms, alkoxy of 1 to 6 carbon atoms, or hydroxy; more preferrably, the alkyl groups have 1 to 4 carbon atoms and the alkoxy groups have 1 to 3 carbon atoms;

2~3'~

Rl is alkyl of 6 to 10 carbon atoms or cycloalkyl of 5 to 7 carbon atoms, most preferrably Rl is alkyl of 6 to 10 carbon atoms, optionally alpha branched;
and R3 is alkyl of 1 to 6 carbon atoms, more preferrably R3 is branched chain alkyl of 3 to 6 carbon atoms.
The prefe~red compounds containing the benzothienyl substituents are those of formula 6:
OX
0~o N N~RI
X ~ ~;X

Z Z

10 in which X, Y and Z are, independently, alkyl of 1 to 4 carbon atoms, alkoxy of 1 to 4 carbon atoms or hydroxy;
and R1 is alkyl of 1 to 18 carbon atoms;
or a pharmaceutically acceptable salt thereof.
The compounds of this invention are prepared by conversion of 2,2-dimethyl-1,3-dioxane-4,6-dione to the corresponding S-bis-(methylthio) methylene derivative with carbon disulfide and methyl iodide in dimethylsulfoxide in the presence of a base such as triethylamine, followed by sequential displacement of the methylthio groups with the desired amines, thusly:

5 -- - ~

~X O~ O
cs2 ~ Et3N + CH3I ' ~
MeS SMe ~J
HN ~ X Y
r~

The following examples illustrate without limitation the preparation of 5 representative compounds of this invention.

~çthod A
Exa~le 1 ~-rrr~,~-Bi~(1.1-dimethylethvl)-4-hvdroxy,phenvllaminolrhexvl-L4-~uinolinylmetbvl)a~olmethvlenel-2.2-dimethvl-1.3-dioxane-4.6-dione p~e 1 To a solution of 6.4 g (25.8 mmol) of 5-[bis(methylthio)methylene]-2,2-dimethyl-1,3-dioxane-4,6-dione and 4.84 g (56.0 mmol) of sodium bicarbonate in 10 mL of degassed DMSO was added 10.0 g (36.0 mmol) of 3,5-di-t-butyl-4-hydroxyaniline hydrochloride in 30 mL of degassed DMSO over a 5 hour period at room temperature. Stirring was continued for an additional 19 hours. The reaction mixture was poured into cold H2O and the product filtered. The solid was dried and dissolved in ethyl acetate and filtered again. The solvent was removed at reduced 2 E377~
- 6 - ~ . . s .

pressure and the residue submitted to a column chromatography on silica gel (3: 1 to 2: 1 hexane-ethyl acetate) to yield 9.8 g (90%) of a solid that was used without further purification.

Procedure 2 To a solution of 3.05 mL (98.8 mmol) of hexylamine and 16.5 mL (33 mmol) of 2N HCI in 25 mL of methanol was dissolved 2.59 g (16.5 mmol) of 4-quinolinecarboxaldehyde. To this mixture was added 0.62 g (9.9 mmol) of sodium cyanoborohydride. After 1 hour the pH was lowered to 7.8 with 2N HCI. This stirred at room temperature for 1 hour. The solvents were removed at reduced pressure and the residue was added to 30 mL of H2O and the solution was acidified to pH2 withconcentrated HCI. This solution was washed 3 X 50 mL of diethyl ether and then it was made basic with NaOH to pH12. The aqueous solution was extracted 3 times with 50 rnL of CHC13 which extracts were combined, dried (Na2SO4) and the solvent removed at reduced pressure. Column chromatography of the residue on silica gel (90:10 ethyl acetate - hexane to 5:95 triethylamine-ethyl acetate) yielded 2.5 g (64%) of an oil which was used without further puri~lcation.

Procedur~ 3 To a solution of 1.2 g (2.86 mmol) of the compound from Method A, P'rocedure 1 in 30 mL of acetonitrile was added 0.73 g (3.0 mmol) of the amine from Method A, Procedure 2, 0.4 mL (2.86 mmol) of triethylamine and 0.47 g (1.57 mmol) of mercuric sulfate. The mixture was allowed to stir at reflux for 4 hours. The solution was cooled, diluted with ethyl acetate and filtered through Celite E9. The solvents were removed at reduced pressure and column chromatography of the residue on silica gel (80:20 ethyl acetate - hexanes to 100% ethyl acetate) yielded 0.52 g (46%) of a yellow powder. This was recrystallized from ethyl acetate - hexanes to yield the title compound as a yellow solid (mp 127-133C). IR (KBr): 3420, 3222, 2950, 2868, 1612 1565, 1507, 1462, 1429, 1381, 1360, 1251, 1229, 1198, 1161, 1113, 1089, 1021, 922, 883, 782 and 762 cm~ H NMR (400 MHz, CDC13): ~ 9.60 (br s, lH), 8.90 (d, lH, J = 4.48 Hz), 8.16 (d, lH, J = 8.36), 7.74-7.61 (m, 3H), 7.55 (t, lH, J
= 7.28 Hz), 6.71 (s, 2H), 5.22 (s, lH), 4.90 (s, 2H), 3.23 (br s, 2H), 1.67 (br s, 8H), 1.22 (br s, 24H), 0.83 (t, 3H, J = 6.76 Hz).

Elemental analysis for C37H4gN3Os Calc'd: C, 72.17; H, 8.02; N, 6.82 Found: C, 71.78; H, 8.00; N, 6.60 S Method 1~
E2~1e 2 ~-~rf2~4~Dimeth~xvr~
uuino~ nethyl)aminQlmethv~ ~thv~ dioxan~ ~-diQD
Pr~cedure 1 To a solution containing 2.0 g (8.05 mmol) of 5-[bis(methylthio)methylene]-2,2-dimethyl- 1,3-dioxane-4,6-dione 1 in 40 mL of t-butanol, was added 1.23 g (8.05 mmol) of 2,4-dimethoxyaniline. The reaction mixture was allowed to stir at reflux for 24 hours. The mixture was cooled to room temperature and diluted with hexanes. The solid was filtered and used without further purification. Isolated: 2.3 g, 81% yield.

procedure 2 To a solution of 0.84 g (2.38 mmol) of the product from Method B, Procedure 1, was added 30 mL of acetonitrile, 0.33 g (1.31 mmol) of mercuric sulfate, 0.33 mL
(2.38 mmol) of triethylamine and 0.64 g (2.6 mmol) of the amine synthesized in Method A, Procedure 2. The reaction mixture was allowed to stir at reflux for 1.5 hours. The rnixture was then cooled to room temperature, diluted with ethyl acetate and filtered through Celite(3~. The solvent was removed at reduced pressure and column chromatography of the residue on silica gel (ethyl acetate to 90:10 ethyl acetate -ethanol) yielded a solid which after recrystallization from diisopropyl ether yielded 1.08 g (83%) of the title compound as a white solid (mp 144-147C). IR (KBr): 3420, 2930, 2855, 1700, 1633, 1571, 1517, 1464, 1386, 1352, 1312, 1204, 1160, 1088, 1031 and 930 cm-l. lH NMR (400 MHz, CDC13): o 8.90 (d, lH, J = 4.36 Hz), 8.8 (br s, lH exchangeable), 8.17 (d, lH, J = 8.08 Hz), 7.76-7.66 (m, 3H), 7.54 (t, lH, J = 7.28 Hz), 6.91 (d, lH, J = 8.52 Hz), 6.33 (d, lH, J = 2.48 Hz), 6.26 (dd, lH, J
= 8.52, 2.52 Hz), 4.94 (br s, 2H), 3.75 (s, 3H), 3.58 (s, 3H), 3.26 (br s, 2H), 1.61 (br s, 8H), 1.25 (br s, 6H), 0.83 (t, 3H, J = 6.84 Hz).

Elemental analysis for C3lH37N3O6 Calc'd: C, 67.99; H, 6.81 Found: C, 67.84; H, 6.91 2~7~?2~

.. .. - .. ~ iJ

Ex~mnl~
~-rrr3.s-si~(l 1 -din~çthyl~-4-hvdroxypbenvllamilLolrh,exvl-(2-auinolinvlmethvl)aminLf~ thvlenel-~dime~l.3-dioxane-4.6-dione s This compound was synthesized using the methodology described in Method A
except 2-quinolinecarboxaldehyde was substituted for 4-quinolinecarboxaldehyde to yield the title compound as a white solid (mp 161-163C).

10Elemental analysis for C37H4gN3Os Calc'd: C, 72.17; H, 8.02 Found: C, 71.91; H, 8.09 F,~ e 4 155-rr(2,4-Dimethoxvpheny~ nolrb*xvl-(2-ouinolinylmethvl)amillolmethvlenel-2.2 dimethvl-1.3-dioxane-4.6-dione This compound was synthesized using the methodology described in Method B
except 2-quinolinecarboxaldehyde was substituted for 4-quinolinecarboxaldehyde to yield the title compound as a light brown solid (mp 106- 109C).

Elemental analysis for C3lH37N3O6 Calc'd: C, 67.99; H, 6.81 Found: C, 67.54; H, 6.82 Exam~
5-rr~2,4-Dim~h~hel~yl~am~-r~ hvlhexvl)~2-~ui~Qlinvlmethvl~amjnolm~h~ene~ -dio~e-4~dione This compound was synthesized using the methodology described in Method A
except 2-aminoheptane was used in place of hexylamine and 2-quinolinecarboxaldehyde was used instead of 4-quinolinecarboxaldehyde to yield the title compound as a solid (mp 148-151C).

Elemental analysis for C32H3sNso6 Calc'd: C, 58.43; H, 7.00; N, 7.48 Found: C, 58.10; H, 7.13; N, 7.29 Exaln~ 6 5-rrr3.5-P~is~ -dimethvlethvl)-4-hvdroxvDhenyllam jaQlr(1 methvlh ~ dime~hy1-1.3-d(~ L~-diorL

This compound was synthesized using the methodology described in Method A
except 2-aminoheptane was used in place of hexylamine and 2-quinolinecarboxaldehyde was used instead of 4-quinolinecarboxaldehyde to yield the title compound as a solid (mp 124- 128C).

Elemental analysis for C3gHslN3Os Calc'd: C, 72.47; H, 8.16; N, 6.67 Found: C, 72.16; H, 8.04; N, 6.29 F,~amDle 7 S-r~(2,4-Dinle~hoxyDhenvl~a~ L~-metbvlhexvl)(4-jl~olinvlmethyl)aminglmethvlenel-2.2-dimethy~-i.3-dioxall,e-4.6-dione This compound was synthesized using the same methodology described in Method B except that 2-aminoheptane was substituted for hexylamine to yield the title 25 compound as a solid (mp 197-202C).

Elemental analysis for C32H39N3o6 Calc'd: C, 68.43; H, 7.00; N, 7.48 Found: C, 68.41; H, 6.89; N, 7.48 ";; ,}

This compound was synthesized using the same me~hodology described in Method A except that 2-aminoheptane was substituted for hexylamine to yicld the title compound as a solid (mp 147-151C).

iO Elemental analysis for C3gH51N3O5 CalG'd: C, 72.46; H, 8.16; N, 6.67 Found: C, 72.11; H, B.21; N, 6.35 E,xample ,~

5~rrr3.5-Bi~ imethvlethYl)~4-hvdroxvghenvllamirlolrkexyl-(3 nolinvlmetllv~ inolmethylenel-2~2~ methv~ 3-diox~n~-4~6-diQn~

This compound was synthesized using the methodology described in Method A
20 except 3-quinolinecarboxaldehyde was substituted for 4-quinolinecarboxaldehyde to yield a light orange powder (mp 130-136C).

Elemental analysis for C37~gN3Os Calc'd: C, 72.17; H, 8.02; N, 6.82 Found: C, '71.74; H, 8.07; N, 6.81 k~E~
Exam~,ç IQ

~r(2~4-Dimethoxvplhenvlamino~!r~ .2-~linlÇlhYlP~X~
thienvlmetllYllheJ2~vlamil~Qlmetlly~ L-Z.2-~ ethYl-L~-diQ~.nne-4.Ç

Procedure 1 To a solution of 50.4 g (0.6 mol) of thiopllene and 72.3 g (0.6 mol) of pivaloylchloride in 500 mL of benzene at 0C was added 70.~ mL (0.6 mol) of SnC14 over a 2~7~

0.75 hour period. The solution stirred at 0C for 0.5 hours then at room temperature for 2 hours. The reaction was quenched with 100 mL of 10% HCI and the organic layer was separated, washed twice with H2O, then dried (MgSO4) and the solvents were removed at reduced pressure. Distillation under vac~uum (1.2 mm Hg) at 71-73C
yielded 67.0 g (5S%) of an oil. IR (film) 3090, 2975, 1640, 1481, 1411, 1363, 1347, 1276, 1178, 1059, 908, 851 and 718 cm-l. IH NMR (200 MHz, CDC13): ~7.83 (d, lH, J = 4.0 Hz), 7.62 (dd, IH, J = 4.0, 3.8 Hz), 7.17 (dd, lH, J = 4.0, 3.8 Hz),1.43 (s, 9H).

Pro~dure 2 To a solution of 67.0 g (0.4 mol) of ketone from Method C, Procedure 1 (thien-2-yl, tert-butyl ketone) in 250 mL of ethanol was added 48.0 mL (1.0 mol) hydrazine.
The reaction mixture was allowed to reflux for 10 days. The solvents were then removed at reduced pressure. The residue was added to 300 mL of toluene and 45 g(0.4 mol) of potassium -butoxide was added. The reaction was heated slowly (exothermic) then taken to reflux for 3.5 hours. The reaction mixture was cooled to room temperature and added to H2O. The layers were separated and the aqueous layer was extracted with diethyl ether. The combined organic layers were dried (MgSO4)and the solvents removed at reduced pressure. Distillation at 16-18 mm Hg yielded 60 gms (98%) of a liquid (B.P. 86-89C). IR (film) 2942, 1478, 1469, 1425, 1387, 1357, 1232, 1194, 1178, 1107, 1072, 1039, 848, 819 and 682 cm-l. IH NMR (80 MHz, CDC13): ~ 7.30 (m, 3H), 2.65 (s, 2H), 0.98 (s, 9H).

procedure 3 To a solution of 11.0 mL (0.14 mol) of dimethylformamide in 15 mL of dichloroethane at 0C was added 13.2 mL (0.14 mol) o~;phosphorous oxychloride over a 0.5 hour period. At 0C a solution 20 g (0.13 molj of the 2-neopentylthiophene30 (Method C, Procedure 2) in 40 mL of ethylene dichloride was added over a 1 hour period. The reaction mixture was allowed to warm to room temperature over 0.5 hours then to reflux for 2 hours. The solution was cooled when 96 g of NaOAc-3 H2O in 200 mL of H2O was added and stirred for 10 minutes. The layers were separated and the aqueous layer was extracted twice with diethyl ether. The combined organic layers 35 were washed with saturated aqueous K2C03, then dried (MgSO4) and concen~rated at reduced pressule. Distillatioll of the residue at ().6 mm Hg yielded 19.6 g ~83%) of an oi3 (BP. 107C). This was used wi~hout further cl-arac~erization.

To a solution of 19.6 g (0.11 mol) of the aldehyde fiom Methocl C, Procedure 3 (5-(2,2-siimethylpropyl)-2-thienyl-carboxaldehyde) in 70 mL of benzene was added16.0 mL ~0.11 mol) of 1-aminoheptane and a crystal of p-toluenesulfonic acid. The reaction mixture was allowed to reflux for 16 hours using a Dean-Stark trap. Thesolution was cooled to room temperature and the solvent was remo~ed at reduced pressure. The residue was added to 250 mI. of dry 'I~IF and HCI gas was bubbled in ~or 10 minutes. The mixture was cooled to 0C and 6.72 g (0.11 mol) of sodium cyanoborohydride in 50 mL of methanol was slowly added. The reaction mixh~re wasallowed to stir at 0C for 0.5 hours then at room temperature ~or 17 hours. The mixture was poured into 150 mL of 0.5 N NaOH and extracted twice with diethyl ether. Thecombined organic layers were dried (Na2S04) and condensed at reduced pressure.
Distillation at 0.4 mm Hg yielded 24.7 g (82%~ of an oil (B.P. 153-155C). IR (film) 3075, 2928, 2854, 1460, 1362, 1238, l l l l, 800 and 739 cm-l. IH NMR (400 MHz, CDCl3): ~ 6.68 (d, IH, J = 6.0 Hz~, 6.54 (d, lH, J = 6.0 Hz), 3.80 (s, 2H), 2.62 (m, 4H), 1.60 (br s, lH), 1.48 (m, 2H), 1.28 ~m, 8H), 0.g4 (s, 9H), 0.88 (t, 3H, J = 5.8 Hz~.

~,~

To a solution containing 2.0 g (8.05 mmol) of 5-[bis(methylthio)methylene]-2,2-dirrlethyl-1,3-dioxane-4,6-dione in 40 mL of -butanol, was added 1.23 g (8.05 mmol) of 2,4-dimethoxyaniline. The reaction mixture was allowed to stir at reflux for 24 hours. The mixture was cooled to room temperature and diluted with hexanes. The solid was filtered and used without further purific.ltion. Isolated: 2.3 g, 81~7/o yield.

To a solution of 0.62 g (1.15 mmol) of the compound from Method C, Procedure S in 10 mL of acetonitrile was added 0.'19 g (1.75 mmol) of the amine from Method C, Procedure 4, 0.31 g (1.()5 mmol) of HgSO4 and 0.25 mL (1.75 mmol) of triethylamine. This reaction mixture was allowed to reflux ~or 18 hours. The mixture 2~

was cooled, diluted with ethyl acetate and filtered through Celite~. The solvent was removed at reduced pressure and the residue was chromatographed on silica gel (2: 1 hexanes-ethyl acetate to 1:1 hexanes - ethyl acetate) to yield 0.73 g (71%) of the title compound as a solid (mp 69-72C). IR (KBr) 3420, 32~10, 2955, 2860, 1702, 1634, 1571, 1512, 1456, 1390, 1367, 1312, 1208, 1160, 1084, 1038, 932, and 890 cm~1.
lH NMR (400 MHz, CDC13): ~ 7.06 (d, lH, J = 8.72 Hz), 6.85 (d, lH, J = 3.40 Hz), 6.61 (d, lH, J = 3.40 Hz), 6.47 (d, lH, J = 2.56 Hz), 6.34 (dd, lH, J = 8.72, 2.56 Hz), 4.57 (s, 2H), 3.81 (s, 3H), 3.77 (s, 3H), 2.96 (m, 2H), 1.7Q (br s, 6H), 1.55 (m, 2H), 1.17 lm, 8H), 0.95 (s, 9H), 0.84 (t, 3H, J = 6.87 Hz).
Elemental analysis for C32H46N206S
Calc'd: C, 65.50; H, 7.90; N, 4.77 Found: C, 65.73; H, 7.97; N, 4.74 ME;T~QD D

s-rrr3.s-si~ dime~ h~lroxvph~ laminolrr5-(2~2-dime~_ylpropvl)-2-thienylmethy11hevtylaminQlmethylenel-2.2-dimethyl-L~-diox~ ~ç~dioL

P,ru~edu~r,el To a solution of 6.4 g (25.8 mmol) of 5-[bis(methylthio)methylene]-2,2-25 dimethyl-1,3-dioxane-4,6-dione and 4.84 g (56.0 mmol) of sodium bicarbonate in 10 mL of degassed DMSO was added 10.0 g (36.0 mmol) of 3,5-di-t-butyl-4-hydroxyaniline hydrochloride in 30 mL of degassed DMSO over a 5 hour period at room tempera.ure. Stirring was continued for an additional 19 hours. The reaction mixture was poured into cold H2O and the product filtered. The solid was dried and 30 dissolved in ethyl acetate and filtered again. The solvent was removed at reduced pressure and the residue submitted to a column chromatography on silica gel (3: 1 to 2: 1 hexane-ethyl acetate) to yield 9.8 g (90%) of a solid that was used without further purification.

- 14- - . . .

PLcedure ~
To a solution of 0.58 g (2.08 mmol) of the amine synthesized in Method C, Procedure 4 in 20 mL of CH3CN was added 0.84 g of th~e compound from Method D, Procedure 1, 0.29 g (0.99 mmol) of HgSO4 and 0.2 g (1.98 mmol) of triethylamine.The solution was allowed to reflux for 18 hours. The reaction mixture was cooled to room temperature, diluted with ethyl acetate and filtered through Celite~. The solvents were removed at reduced pressure and the residue was chromatographed on silica gel (3: 1 hexanes - ethyl acetate to 1: 1 hexanes - ethyl acetate) to yield after recrystallization (hexanes - ethyl acetate) 1.0 g (77%) of the title compound as a pale yellow solid (mp 178-179C). IR (KBr) 3405, 2950, 2859, 1696, 1623, 1573, 1462, 1432, 1383, 1361, 1233, 1204, 1115, 1088, 931 and 800 cm-l. lH NMR (400 MHz, CDC13):
6.93 (s, 2H), 6.85 (d, lH, J = 3.32 Hz), 6.59 (d, lH, J = 3.32 Hz), 5.23 (br s, lH), 4.56 (s, 2H), 3.05 (m, 2H), 2.62 (s, 2H), 1.72-1.55 (m, 8H), 1.38 (s, 18H), 1.34-1.09 (m, 8H), 0.94 (s, 9H), 0.85 (t, 3H, J = 6.9 Hz).

Elemental analysis for C3sH4sN2Oss Calc'd: C, 69.65; H, 8.93; N, 4.28 Found: C, 69.76; H, 9.05; N, 4.12 Exam~le 12 meJ~hQ~2heny~ ~ ~imel~hvl~rQrlyl)~2 thje~y~methy~ h~exyl)amjnnlm~h~el-2,2-dinl,e~
diox~-4.6-dione This compound was synthesized using the same methodology as in Method C
except 2-aminoheptane was substituted for 1-aminoheptane to yield 1.37 g (81%) of a white powder (mp 152-153C).
Elernental analysis for C32H46N2o6s Calc'd: C, 65.50; H, 7.90; N, 4.77 Found: C, 65.88; H, 8.00; N, 4.66 7.~

~yamL~le 13 5-Frr3.5~ dimethvlethyl)-4-hvdroxvvhenyl~m~_lrr$-(2.~-dimethy~opvl)-2-thienylmethvll~l-methvlhe S dim~thvl l.3-dioxalle-4.6-~J.Q~

This compound was synthesized using the same methodology as in Method FDexcept 2-aminoheptane was substituted for 1-aminoheptane in Method C,Procedure4 to yield 2.0 g (85%) of a white powder (mp 184-185C).
Elemental analysis for C3gHsgN2OsS
Calc'd: C, 69.69; H, 8.92; N, 4.28 Found: C, 69.56; H, 8.83; N, 4.34 Example 14 5-rrCvcloll,~e2~y~ ~thvl~ro~vl)-2-thienvlmethvllaminol-r2.4-dimethQ~YIlh~olmethYlenel-2 2-dimethyl-1.3-nioxan~-4.6-dj ,o, rLe This compound was synthesized using the same methodology as in Method C
except cyclohexylamine was substituted for 1-aminoheptane to yield 1.21 g (71 %) of an off-white powder (mp 143-145C).

Elemental analysis for C31H42N2O6S
Calc'd: C, 65.24; H, 7.42; N, 4.91 Found: C, 65.61; H, 7.57; N, 4.60 2~ o~

3_- A ' Metho~L~
E~ljam~ 15 S-r(Benzorllltl~jophen-3.vlmethxl)-hexvlanl,~Lo)-(3,5-di-te~.butyl.4.
5 hvdrs~xvphenvlamino)methylenel.2.2-dimethyl-rl,~ldioxane-4.6-dione Procedurç 1 To a solution of 7.0 g (52.16 mmol) of thianapthene in 315 mL of anhydrous diethyl ether at -78 C was added 34 mL of 2.5 M n-butyllithium dropwise. ~fter 0.5 hours at -78C, the solution was warmed slowly to 0C and a solution of 6.4 mL
(52.16 mmol) of N-methylformanilide in 10 mL of diethyl ether was added dropwise.
After 1.5 hours at 0C, the reaction mixture was slowly brought to reflux for 1 hour.
The mixture was cooled to room temperature and poured into 30 mL of 3N HCI. The layers were separated and the aqueous layer was extracted 3 times with 30 mL of diethyl ether. The combined ether extracts were then washed with lN HCI (30 mL) and once with saturated aqueous NaHCO3 (30 mL) then dried (MgSO4) and the solvents removed at reduced pressure. The residue was dissolved in 15 mL of ethanol and 4.5 mL of saturated sodium bisulfite was added. After thorough mixing, the solution was allowed to stand at room temperature for 20 minutes. The white precipitate was filtered and washed three times with diethyl ether. After drying under vacuum, the solid was dissolved in H2O, cooled to O C and saturated Na2CO3 was added. This aqueous layer was extracted with ethyl acetate which was dried (MgSO4), filtered, and the solvents were removed at reduced pressure. Column chromatography of the residue on silica gel (275 g silica gel, 93% hexanes - 7% ethyl acetate to 89%
hexanes - 11% ethyl acetate) gave after crystallization (diethyl ether-hexanes) 1.32 g of a solid which was used without further purification or characterization.

Proce To a solution of 0.97 g (5.98 mmol) of 2-benzo[b]thiophenecarboxaldehyde, from Method E, Procedure 1, in 10 mL of methanol was added 1.81 g (17.9 mmol) ofhexylamine and then methanol saturated with gaseous HCI until the pH was 7. Sodium cyanoborohydride, 0.25g (4.18 mmol), was added as a solid and stirred at room temperature for 18 hours. The pH of the solution was lowered to 2 with concentrated HCI and the methanol was removed at reduced pressure. H2O was added to the residue 2~

. ~ , which was washed with diethyl ether. The aqueous layer was made basic with solidKOH and the solution was extracted twice with ethyl acetate. The combined ethyl acetate layers were dried (Na2SO4) and the solvents removed under reduced pressure.
Column chromatography of the residue (120 g silica gel,75% EtOAc-hexanes) yielded S 0.55 g (37%) of a yellow oil. NMR (200 MHz, CDC13) ~ 7.8-7.63 (m 2H), 7.36-7.20 (m, 2H), 7.12 (s, lH), 4.06 (s, 2H), 2.66 (t, 2H, J=6.4 Hz), 1.58-1.22 (m, 8H), 0.90 (t, 3H, J=6.6 Hz).

Proc~dur~ 3 To a solution of 6.4 g (25.8 mmol) of 5-[bis(methylthio)methylene]-2,2-dimethyl-1,3-dioxane-4,6-dione and 4.84 g (56.0 mmol) of sodium bicarbonate in 10 mL of degassed DMSO was added 10.0 g (36.0 mmol) of 3,5-di-t-butyl-4-hydroxyaniline hydrochloride in 30 mL of degassed DMSO over a S hour period at room temperature. Stirring was continued for an additional 19 hours. The reaction mixture was poured into cold H2O and the product filtered. The solid was dried and dissolved in ethyl acetate and filtered again. The solvent was removed at reduced pressure and the residue submitted to a column chromatography on silica gel (3: 1 to 2: 1 hexanes-ethyl acetate) to yield 9.8 g (90%) of a solid that was used without further purification.

Procedure 4 To a solution of 0.55 g (2.22 mmol) of the amine from Method E, Procedure 2 in 20 mL CH3CN was added 0.94 g (2.22 mmol) of the compound from Method E, Procedure 3, 0.22 g (2.22 mmol) of triethylamine and 0.36 g (1.22 mmol) of HgSO4.
The reaction mixture was allowed to reflux for 18 hours. After cooling to room temperature, the reaction mixture was filtered through Celite g~ and the solvents were removed at reduced pressure. Column chromatography of the residue (110 gm silicagel, 65:35 hexanes-ethyl acetate) yielded after recrystallization (ethyl acetate-hexanes) 0.95 gm (69%) of a solid (m.p. 155-159C). IR (KBr) 3410, 3220, 2952, 2870, 1688, 1635, 1554, 1432, 1383, 1352, 1251, 1203, 1117, 1089, 929 and 740 cm-l.
lH NMR (400 MHz, CDC13) ~ 7.76 (d, lH, J=7.05 Hz), 7.71 (d, lH, J=6.95 Hz), 7.35 (m, 2H), 7.24 (s, lH), 6.95 (s, lH), 5.23 (s, lH), 4.69 (s, lH), 3.09 (m, 2H), 1.69 (s, 6H), 1.63 (m, 2H), 1.35 (s, 18H), 1.26-1.19 (m, 6H), 0.83 (t, 3H, J=6.9Hz).

2~77 Elemental analysis for C36H48N2O5S
Calc'd: C, 69.64; H, 7.79; N, 4.51 Found: C, 69.81; H, 8.09; N, 4.38 s MethQ~I F
E~mJ21 5-rr(BeDz~[~l~he~ h~x~laminol~(2 4 d_o)~hy~ di~rlh~ 4 Procedure 1 To a solution containing 2.0 g (8.05 mmol) of 5-[bis(methylthio)methylene]-2.2-dimethyl-1,3-dioxane-4,6-dione in 40 mL of t-butanol, was added 1.23 g (8.05mmol) of 2.4-dimethoxyaniline. The reaction mixture was allowed to stir at reflux for 24 hours. The mixture was cooled to room temperature and diluted with hexanes. l he solid was filtered and used without further purification. Isolated: 2.3 g,81 % yield.

P-~

To a solution of 0.76 g (3.08 mmol) of the amine from Method GE Procedure 2, in 15 mL of acetonitrile, was added 0.31 g (3.08 mmol) of triethylamine, 1.08 g (3.08 mmol) of the product from Method F, Procedure 1 and 0.50 g (1.69 mmol) of 25 HgSO4. The reaction mixture was allowed to stir at reflux for 18 hours. The mixture was cooled to room temperature, filtered through Celite~ and the solvents removed at reduced pressure. Column chromatography (120 g silica gel, 65:35 hexanes-ethyl acetate to 50:50 hexanes-ethylacetate) of the residue yielded after recrystallization (ethyl acetate-hexanes) a yellow solid (m.p. 96-99C). IR (KBr) 3430, 2960, 2925, 2860,30 1697, 1632, 1572, 1512, 1456, 1437, 1386, 1347, 1312, 1208, 1158, 1079, 1032,930 and 788 cm-l. IH NMR (400 MHz, CDCL3) ~ 7.79 (d, 2H, J=8.0 Hz), 7.71 (d, lH, J=7.8 Hz), 7.33-7.28 (m, 2H), 7.25 (s, lH), 7.14 (d, lH, J=8.7 Hz), 6.45 (d,lH, J=2.7 Hz), 6.30 (dd, lH, J=2.68, 8.72 Hz), 4.70 (s, 2H), 3.75 (s, 6H), 3.07 (m, 2H), 1.64-1.56 (m, 8H), 1.22-1.12 (m, 6H), 0.80 (t, 3H, J=6.84 Hz).

:'' ~ -.

~, ~ 7....~J~
lg Elemental analysis for C30H36N2o6s Calc'd: C, 65.20; H, 6.57; N, 5.07 Found: C7 64.85; H, 6.58; N, 5.02 ~~
This conlpound was synthesized using the methodology described in Method F except 2-aminoheptane was substituted for hexylamine to yield a white solid (m.p.
167 C).

Elemental analysis for C31H38N2O6S
Calc'd: C, 65.70; H, 6.76; N, 4.94 Found: C, 65.64; H, 6.67; N, 4.84 Ex~mple 1 2~ 5- uBenzQr~~io~

This compound was synthesized using the methodology described in Method E except 2-aminoheptane was substituted for hexylamine to yield a white solid (m.p.
195-197 C).

Elemental analysis for C37HsoN2oss Calc'd: C, 70.00; ~1, 7.94; N, 4.41 Found: C, 70.21; ~1, 7.85; N, 4.46 The ability of the compounds of this invention to inhibil acyl-coenzyme A:
cholesterol acyltransferase was established by initially showing that they inhibited intracellular cholesterol esterification by subjecting them to the standard experimental test procedure of Ross et al., J. Biol. Chem. 252 815 (1984). The results of these siudies are presented in:

2~

Table I

Example%Inhib. (Concentra~
1 69 (25) 2.96 3 55 (25) 2.33 4 72 (25) 5.16 90 (25) 2.42 6 95 (25) 1.55 7 51(25) 9.41 8 97 (25) 1.65 9 98 (25) 1.35 94 (25) 2.96 11 ~ ' 15 12 97 (25) 13 - 0.64 14 95 (25) 98 (25) 0.38 16 94 (25) 0.62 20 17 97 (25) 0.35 18 98 (25) 0.31 Representative compounds were further tested in vivo to establish the percent inhibition of cholesterol absorption. In this study, normal rats were dosed (oral gavage) with 14C-cholesterol plus the test compound. Blood samples taken exactly six hours later were analyzed and the percent inhibition of cholesterol absorption was calculated as shown in:

Table II
In Vivo Testing 14C-Cholesterol Absorption in Normal Rats % Inhibidon ExampleDose mg~c~ of Absorption 2~

In addition, the product of Example 1 was studied in vivo in the cholesterol-cholic acid fed rat to determine the percent decrease of cholesterol in their plasma This study involves rats which are, prior to testing, trained for one week to eat over a four 5 hour time period each day. Upon initiation of the experiment, the rats diet issupplemented wilh 1.0 percent cholesterol and 0.25 percent cholic acid. The rats are dosed with the test compound by oral gavage just prior to and just following the four hour feeding period. This is repeated for four days. On the fifth day, the rats are sacrificed and the total plasma cholesterol content is determined. The percent decrease 10 in elevated plasma cholesterol levels is calculated in comparison with normal-fed controls. The compound of Example 1 at 10 mg/kg resulted in a 50 percent decrease in plasma cholesterol.

From these data, the ability of the compounds to inhibit ACAT is clearly 15 established. Hence, the compounds of this invention are useful in the treatment of those disease states which are amenable to treatment by reduction of the rate ofcholesterol esterification, the rate of accumulation and deposits of cholesteryl esters on arterial walls and the rate of formation of atheromatous lesions. As such, the anti-atherosclerotic agents of this invention may be administered to a mammal in need of 20 intracellular cholesteryl ester concentration reduction orally or parenterally in an amount sufficient to inhibit ACAT catalysis of cholesterol esterification.

The compounds of this invention may be administered by themselves or in combination with pharmaceutically acceptable liquid or solid carriers. Oral 25 administration in conventional formulations as tablets, capsules, powders, or suspensions is preferred.

A solid carrier can include one or more substances which may also act as flavoring agents, lubricants, solubilisers, suspending agents, fillers, glidants, 30 compression aids, binders or tablet-disintegrating agents; it can also be an encapsulating material. In powders, the carrier is a finely divided solid which is in admixture with the finely divided active ingredient. In tablets, the active ingredient is mixed with a carrier having the necessary compression properties in suitable proportions and compacted in the shape and size desired. The powders and tablets preferably contain up to 99% of 35 the active ingredient. Suitable solid carriers include, for example, calcium phosphate, magnesium stearate, talc, sugars, lactose, dextrin, starch, gelatin, cellulose, methyl 7 ~~~

cellulose, sodium carboxymethyl cellulose, polyvinylpyrrolidine, low melting waxes and ion exchange resins.

Liquid carriers are used in preparing solutions, suspensions, emulsions, 5 syrups, elixirs and pressurized compositions. The active ingredient can be dissolved or suspended in a pharmaceutically acceptable liquid carrier such as water, an organic solvent, a mixture of both of pharmaceutically acceptable oils or fats. The liquid carrier can contain other suitable pharmaceutical additives such as solubilisers, emulsifiers, buffers, preservatives, sweeteners, flavoring agents, suspending agents, thickening 10 agents, colors, viscosity regulators, stabilisers or osmo-regulators. Suitable examples of liquid carriers for oral and parenteral administration include water (particularly containing additives as above, e.g. cellulose derivatives, preferably sodium carboxymethyl cellulose solution), alcohols (including monohydric alcohols and polyhydric alcohols, e.g., glycols) and their derivatives, and oils (e.g. fractionated 15 coconut oil and arachis oil). For parenteral administration, the carrier can also be an oil ester such as ethyl oleate and isopropyl myristate. Sterile liquid carriers are used in sterile liquid form compositions for parenteral administration. The liquid carrier for pressurized compositions can be halogenated hydrocarbon or other pharmaceutically acceptable propellent.
Liquid pharmaceutical compositions which are sterile solutions or suspensions can be utilized by, for example, intramuscular intraperitoneal or subcutaneous injection.
Sterile solutions can also be administered intravenously. When the compound is orally active, it can be administered orally either in liquid or solid composition form.
Preferably, the pharmaceutical composition is in unit dosage form, e.g. as tablets or capsules. In such form, the composition is sub-divided in unit dose containing appropriate quantities of the active ingredient; the unit dosage forms can be packaged compositions, for example, packeted powders, vials, ampoules, prefilled30 syringes or sachets containing liquids. The unit dosage form can be, for example, a capsule or tablet itself, or it can be the appropriate number of any such compositions in package form.

The dosage to be used in the treatment of a specific 35 hypercholesterolemic/atherosclerotic condition must be subjectively determined by the ,, - - , .

~ ~ ... . .

7.~3,^~

attending physician. The variables involved include the extent of the disease state, size, age and response patte~n of the patient.

Claims

THE EMBODIMENTS OF THE INVENTION IN WHICH AN EXCLUSIVE
PROPERTY OR PRIVILEGE IS CLAIMED ARE DEFINED AS FOLLOWS:

A compound of the formula:

in which X and Z are, independently, hydrogen, halogen, hydroxy, nitro, cyano, carboxyl, trifluoromethyl, phenyl, amino, alkylamino of 1 to 12 carbon atoms, dialkylamino in which each alkyl group has 1 to 12 carbon atoms, alkyl of 1 to 12 carbon atoms or alkoxy of 1 to 12 carbon atoms;
R1 is hydrogen, alkyl of 1 to 18 carbon atoms, alkenyl of 2 to 18 carbon atoms, cycloalkyl of 5 to 8 carbon atoms, phenyl, benzyl or substituted phenyl or benzyl where the substituents are alkyl of 1 to 12 carbon atoms or alkoxy of 1 to 12 carbon atoms;
R2 is , , or or a pharmaceutically acceptable salt thereof.

A compound of Claim 1 of the formula:

in which X and Z are, independently, alpha branched alkyl of 1 to 6 carbon atoms;
R1 is alkyl of 1 to 18 carbon atoms;
and R2 is 2-, 3- or 4-quinolinyl;
or a pharmaceudcally acceptable salt thereof.

A compound of Claim 1 which is 5-[[[3,5-bis(1,1-dimethylethyl)-4-hydroxyphenyl]amino][hexyl-(4-quinolinylmethyl)amino]methylene]-2,2-dimethyl-1,3-dioxane-4,6-dione, or a pharmaceutically acceptable salt thereof.

A compound of Claim 1 which is 5-[[[3,5-bis(1,1-dimethylethyl)-4-hydroxyphenyl]amino][hexyl-(3-quinolinylmethyl)ammino]methylene]-2,2-dimethyl-

1,3-dioxane-4,6-dione, or a pharmaceutically acceptable salt thereof.

A compound of Claim 1 of the formula:

.

in which X, Y and Z are, independently, hydrogen, alkyl of 1 to 6 carbon atoms, alkoxy of 1 to 6 carbon atoms, or hydroxy;

R1 is alkyl of 6 to 10 carbon atoms or cycloalkyl of 5 to 7 carbon atoms;

R3 is alkyl of 1 to 6 carbon atoms.

A compound of Claim 1 which is 5-[[3,5-bis(1,1-dimethylethyl)-4-hydroxyphenylamino][[5-(2,2-dimethylpropyl)-2-thienylmethyl]heptylamino]-methylene]-2,2-dimethyl-1,3-dioxane-4,6-dione.

A compound of Claim 1 which is 5-[[3,5-bis(1,1-dimethylethyl)-4-hydroxyphenylamino][[5-(2,2-dimethylpropyl)-2-thienylmethyl](1-methylhexyl)-amino]methylene]-2,2-dimethyl-1,3-dioxane-4,6-dione.

A compound of Claim 1 of the formula:

in which X, Y and Z are, independently, alkyl of 1 to 4 carbon atoms, alkoxy of 1 to 4 carbon atoms or hydroxy;
and R1 is alkyl of 1 to 18 carbon atoms;
or a pharmaceutically acceptable salt thereo A compound of Claim 1 which is 5-[[benzo[b]thiophen-2-ylmethyl-(1-methyl-hexyl)amino]-(3,5-di-tert-butyl-4-hydroxy-phenylamino)methylene]-2,2-dimethyl-[1,3]dioxane-4,6-dione, or a pharmaceutically acceptable salt thereof.