United States Patent [191
`
`[11] Patent Number:
`
`4,716,175
`
`I-Ioefle et al.
`[45] Date of Patent:
`Dec. 29, 1987
`
`
`[54] SATURATED FATIT ACID AMIDES AS
`
`[56]
`
`References Cited
`
`ACYLTRANSFERASE
`
`4,410,597 10/1933 Torok et :1}.
`
`545/337
`
`'
`
`[75]
`
`Inventors: Milton L. Hoefle, Ann Arbor; Ann
`H°'m= Dexter; 3"” 9- R‘'“'« Am
`Arbor, all OfM1Ch.
`[73] Assignec: wane;-.Lambe1-1; Company, Mon-is
`Plains, NJ.
`
`[21] Appl. No.: 17,960
`.
`1221 mv
`[51]
`Int. CL4 ................... .. A61K 31/44; ASIK 31/15;
`AGIK 31/165; C0713 213/36; CD7C 103/19
`[52] us. (:1.
`514/357; 514/506;
`545/337; 545/335; 544/302; 544/311; 544/315;
`544/319; 544/335; 564/189; 564/190; 564/204;
`564/219
`546/336, 337; 564/189,
`564/190, 219, 204; 514/357
`
`[58] Field of Search
`
`OTHER PUBLICATIONS
`Chem. Abstracts; Boyles et a1., vol. 94, No. 121134w
`
`Chem. Abstracts; Volkenshtein et al., vol. 104, 199590111
`(1986).
`
`Primary ExamI’ner—-Richard A. Schwartz
`A '3 IE3:
`' —J.Ri'ht
`...:,'::"::m_:.,:;F.
`TR ,
`
`CT
`ABS
`[57]
`Certain substituted amides of saturated fatty acids are
`potent inhibitors of the enzyme acyl-CoA:cholesterol
`acyltransferase and are thus useful agents for inhibiting
`the intestinal absorption of cholesterol.
`
`7 Claims, No Drawings
`
`Iufll
`
`PENN EX. 2195
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`CFAD V. UPENN
`lPR20l5-01836
`
`

`
`1
`
`4,716,175
`
`SATURATED FATTY ACID AMIDES AS
`INHIBITORS OF ACYL-COA:CI-IOLESTEROL
`ACYLTRANSFERASE
`
`BACKGROUND OF THE INVENTION
`
`10
`
`I5
`
`20
`
`25
`
`30
`
`This invention relat to chemical compounds having
`pharmacological activity, to pharmaceutical composi-
`tions which include these compounds, and to a pharma-
`ceutical method of treatment. More particularly, this
`invention concerns certa.in substituted amides of ct-sub-
`stituted or a,t1-disubstituted alkanoic and alkenoic acids
`which inhibit acyl-coenzyine A:cholesterol acyltrans-
`ferase (ACAT), pharmaceutical compositions contain-
`ing these compounds, and a method of inhibiting intesti-
`nal absorption of cholesterol.
`In recent years the role which elevated blood plasma
`levels of cholesterol plays in pathological conditions in
`man has received much attention. Deposits of choles-
`terol in the vascular system have been indicated as caus-
`ative ot‘ a variety of pathological conditions including
`coronary heart disease.
`Initially,
`studies of this problem were directed
`toward finding therapeutic agents which would be cl‘-
`fective in lowering total serum cholesterol levels. It is
`now known that cholesterol is transported in the blood
`in the form of complex particles consisting of a core of
`cholesteryl esters plus triglycerides and an exterior
`consisting primarily of phospholipids and a variety of
`types of protein which are recognized by specific recep-
`tors. For example, it is now known that cholesterol is
`carried to the sites of deposit in blood vessels in the
`form of low density lipoprotein cholesterol (LDL cho-
`lesterol) and away from such sites of deposit in the form
`of high density lipoprotein cholesterol (HDL choles-
`terol).
`Following these discoveries, the search for therapeu-
`tic agents which control serum cholesterol turned to
`finding compounds which are more selective in their
`action; that is, agents which are effective in elevating
`the blood serum levels of HDL cholterol and/or low-
`ering the levels of LDL cholterol. While such agents
`are effective in moderating the levels of serum choles-
`terol, they have little or no effect on controlling the
`absorption of dietary cholesterol into the body through
`the intestinal wall.
`In intestinal mucosa] cells dietary cholesterol is ab-
`sorbed as free cholesterol which must be esterified by
`the action of the enzyme acyl-CoA:cl1olesterol acyl-
`transferase (ACAT) before it can be packaged into the
`chylomicrons which are then released into the blood
`stream. Thus, therapeutic agents which effectively in-
`hibit the action of ACAT prevent the intestinal absorp-
`tion of dietary cholesterol into the blood stream or the 50
`reahsorption of cholesterol which has been previously
`released into the intestine through the body’s own regu-
`latory action.
`
`35
`
`40
`
`45
`
`50
`
`55
`
`65
`
`SUMMARY OF TI-IE INVENTION
`
`The present invention provides a class of compounds
`with ACAT inhibitory activity having the structure
`
`where A is an unbranched hydrocarbon group contain-
`ing from one to twenty carbon atoms and which may
`contain from one to three carbon-carbon double bonds.
`R1 is hydrogen or alkyl of from one to four carbon
`atoms or phenylmethyl and R; is alkyl of from one to
`four carbon atoms or phenylmethyl. Alternatively, R1
`and R2, taken together with the carbon atom to which
`they are attached may form a saturated carbocyclic ring
`of from three to seven carbon atoms.
`B is selected from
`
`(CH2)n
`
`R4
`
`R3 R1
`
`1
`N
`
`N
`
`Rs
`
`Rs
`
`and
`
`CH3
`|
`_cH
`
`N
`
`I
`
`‘x.
`
`I
`
`where n is zero or one, R3, R4, and R5 are independently
`selected from hydrogen, fluorine, chlorine, bromine,
`trifluoromethyl, alkyl of from one to four carbon atoms,
`and alkoxy of from one to four carbon atoms.
`R4; is allroxy of from one to four carbon atoms, and
`R7 and Rs are independently hydrogen or alkoxy of
`from one to four carbon atoms.
`The terms “alkyl” as used throughout this specifica-
`tion and the appended claims means a branched or un-
`branched hydrocarbon grouping derived from a satu-
`rated hydrocarbon by removal of a single hydrogen
`atom. Examples of alkyl groups contemplated as falling
`within the scope of this invention include methyl, ethyl,
`propyl,
`l-methylethyl,
`butyl,
`l-methylpropyl,
`2-
`methylpropyl. and 1, l -dimethylethyl.
`The term “all:oxy" means an alkyl group, as defined
`above, attached to the parent molecular moiety through
`an oxygen atom.
`The term “halogen” contemplates fluorine, chlorine,
`or bromine.
`Those compounds of the present invention in which
`the rt-carbon atom'of the acid portion of the amide is
`only monosubstituted possess an asymmetric center at
`that carbon atom and are capable of existing in two
`enantiomeric forms. Likewise, an asymmetric center
`exists at C1 of the ethyl group in those compounds of
`this invention where the “B” substituent is l-(2-, 3-, or
`4-pyridinyl)ethyl. The present invention contemplates
`all possible optical isomeric forms as well as mixtures
`thereof.
`
`DETAILED DESCRIPTION
`
`The compounds of the present invention provide a
`class of amides of unsubstituted or a,a-disubstituted
`straight-chain acids which are inhibitors of the enzyme
`
`Zofll
`
`PENN EX. 2195
`
`CFAD V. UPENN
`lPR20l5-01836
`
`

`
`4,716,175
`
`3
`acyl—CoA:cl'1olesterol acyltransferase (ACAT) and are
`thus useful as pharmacological agta for inhibiting the
`intestinal absorption of cholesterol.
`The compounds of the present invention are substi-
`tuted with phenylmethyl groups or one or more alkyl
`groups, containing from one to four carbon atoms, on
`the ct-carbon atom of the acid portion of the amide.
`Preferred compounds "of the invention are those in
`which the a-carbon substituents, R1 and Rzare methyl,
`ethyl or phenylmethyl or those compounds where R1
`and R; taken together with the carbon atom to which
`they are attached form a saturated carbocyclic ring of
`Erom three to seven carbon atoms. It has been found, in
`accordance with the present invention, that when one
`or more alkyl groups are attached to the ct-carbon (i.e.
`the carbon atom immediately adjacent to the carbonyl
`function) of the acid residue of the amide compounds of
`this invention, the in vivo ACAT inhibitory activity of
`the compounds is enhanced over the corresponding
`unsubstituted compounds.
`The amide nitrogen of the compounds of this inven-
`tion is substituted with a group selected from phenyl or
`benzyl, either of which may be mono-, di-, or trisubsti-
`tuted with fluorine-, chlorine, bromine, triiluoromethyl,
`alkyl, or alkoxy; mono-, di. or trisubstituted pyrimidin-
`5-yl; or 1-(2-, 3- or 4-pyridinyl)ethyl.
`Preferred compounds of the present invention are
`those in which the alkyl or alltoxy substituenm contain
`one or two
`atoms, i.e. methyl, ethyl, methoxy,
`.' and ethoxy.
`Compounds falling within the scope of the present
`'--'-'invention are emplified by the following:
`' " N-(2,6-Dimethylphenyl)-2,2-dimethyldodecanamide.
`N-(2,6-Diethylphenyl)-2,2-dimethyldoclecanamide.
`N-[2,6-bis(1-Met.hylethy1)phenyi]-2,2-di1nethyl-
`dodecanamide.
`- N-(2-Ethoxy-6-methylphenyl)—2,2-dimethyl-dodecana-
`'
`. mide.
`
`_:-_2-Methyl—N—[2,6-bi.s(l-n:|ethylethyl)phenyl]-tet-
`'
`radecanamide.
`'::(Z)-N-(2,6—Diethylphenyl-2-methyl-9-octadecenamide.
`(Z)-N-{2,6—Diethy1phenyl}-2.2-din1ethyl-9-octadecena-
`mide.
`
`(Z)-N-(2-Methoxy-6-methylpheuyl)-2,2-
`eicosenamide.
`
`din1ethyl—ll-
`
`2,2-Dimethyl-N-(2,4,6-trin1ethoxyphenyl)dodecana-
`mide.
`
`2-Methyl-N-(2,4,6—trimethoxyphenyl)tetradecanamide.
`2-Ethyl-N-(2,4,6—trimethoxypheuyI)tetradecanamide.
`2,2-Dimethyl-N-(2,4-,6-trimethoxyphenyl)tetradecana-
`mide.
`
`2-MethyI-N-(2,4,6—trirnethoxyphenyl)hexadecanan1ide.
`2,2-DimethyI—N-(2,4,6-trimetho xyphenyl)hexadecana-
`mide.
`
`2,2-Dimethyl-N-(2,4,6-trimethoxyphenyl)octadecaoa-
`mide.
`1-Decyl-N-(-2,4,6—trimethoxyphenyl)cyclobutanecar-
`boxamide.
`
`I0
`
`15
`
`20
`
`25
`
`30
`
`35
`
`-10
`
`45
`
`SD
`
`4
`N-(4,6-Dimethoxy-2-phenyl-5-py'rimidinyl)-2,2-dime-
`thyldodecanarnide.
`N.
`
`-(4,6-Dimethoxy-5-pyrintidinyl)-2-rnethylteb
`radecanamide.
`
`N-(4,6-Dimethorny-5-pyrimidinyl)-2-ethyltetradecanm
`mide.
`
`N-(-4,6-Dimethoxy-5-pyrimidinyl)-2,2-dimethyltet-v
`radecanamide.
`
`N-(4.6-Diethoxy-5-pyrimidinyl)-2-methyltetradecanm
`mide.
`
`I-Decyl-N-(4,6-dimethoxypyrimidin-5—yl)cyclopem
`tauecarboxamide.
`(Z)-N-(-4,6-Dirnethoxw 5-py'rimidinyl)-2,2-din1ethyl- l 1-
`eicosenamide.
`
`2-Methyl-N-[1-(2-pyridinyl)ethyl]dodecanamide.
`2-Ethyl-N-[l-(2-pyridinyl)ethyl]dodecanatnide.
`2-Propyl-N-I1-(2-pyridinyI)ethyl]dodecanamide.
`ct-Decyl-N-[l-(2-pyridinyl)ethyl]benzenepropanamide.
`2-Methyl-N-[1-(2-pyridinyl)ethyl]tetra.decanamide.
`2-Ethyl-N [l-(2-py'ridinyl)ethyl]tetradecanamide.
`2-Methyl-N [l-(2-pyridinyl)ethyl]hexadecanamide.
`2,2-Dimethyl-N-[1-(2-pyridinyl)ethyl]hexadeca.uamide.
`The compounds of the present invention are prepared
`by reacting the acid chloride of the appropriate Ct-S1Jb-
`stituted or a,a-disubstituted acid with the desired sub-
`stituted amine in a polar solvent such as tetrahydrofu-=
`ran, chloroform, dimethylformamide, and the like in the
`presence of a tertiary amine acid scavenger such as
`triethylamine.
`The reaction may be carried out at any temperature
`between 0' C. and the boiling point of the solvent, with
`lower temperatures being preferred.
`The reaction is allowed to proceed until analysis of
`the mixture by a means such as chromatography indi-
`cates substantially complete reaction between the acid
`chloride and the substituted amine. Reaction limes may
`vary between about two hours to about 24 hours, de-
`pending upon the particular reagents and reaction tem-
`perature employed.
`Starting materials are known or, if not previously
`known. are prepared by methods well known in the art.
`For example, the starting ct-alkyl-substituted acids are
`prepared by fast converting diethyl malonate to the
`desired alkyl diethyl inalonate and then reacting the
`sodio-salt of the alkyl diethyl malonate with a bromoa1-
`kane using conventional methods. The product of this
`reaction is then hydrolyzed and decarhoxylated by well
`known methods to produce the a-(alkylsubstituted)
`acid. The acid is converted to the acid chloride by
`reaction with thionyl chloride, oxalyl chloride, phos-
`phoryl chloride or the like by conventional methods.
`The starting cr.,cr.-dialykl-substituted acids may be
`prepared by either of two alternative methods. In the
`case where both a-substituents are methyl, the appro-
`priate bromoalltane is reacted with the lithio salt of
`iso-butyric acid or an ester of isobutyric acid to produce
`the desired a,c:-dimethyl acid.
`In the alternative method for preparing the a,a-dial-
`kyl-substituted acids, the appropriate bromoalkane is
`reacted with the sodio salt of diethyl malouate, to pro-
`duce the alkyl-substituted diethyl malonate. This ceteris
`then hydrolyzed to the corresponding alltyl-substituted
`malonic acid and decarboxylated in the conventional
`manner. The resulting monocarboxylic acid is then
`ct-alkylated by first converting the acid to its ct-lithio
`carbanion salt, and then reacting that salt with the ap-
`
`1-Decyl-N-(2,4,6-trimethoxyphenyl)cyclopentanecar-
`boxamide.
`(Z)-2-Methyl-N-
`-(2,4-,6-trimethoxyphenyl)-9-octadecenamide.
`(Z)-2,2-Dimethy1—N-(2,4,6-trimethoxyphenyl}
`octadecenamide.
`
`60
`
`(Z)-2,2-Dimethyl—N-(2,4, 6-trimethoxyphenyl)- I 1-
`eicosenatnide.
`
`N-(4.6-Dimethoxy-5-pyriniidinyl)-2,2-dimethyl-»
`dodecanamide.
`
`3ofll
`
`PENN EX. 2195
`
`CFAD V. UPENN
`lPR20l5-01836
`
`

`
`4,716,175
`
`5
`
`10
`
`I5
`
`20
`
`25
`
`30
`
`6
`TABLE 1-continued
`IC5o
`
`(Micmmolar)
`Compound
`0.40
`
`§—(4,6-dimethoJry-5-pyrirnid.inyl)-2-
`mclhylbetradecanamide
`§—(4.6-dimethoJry-5-pyrimidi.nyl)-2-
`ethyltetradecansrnide
`§—(4,6-dimethoxy-5-pyrimidinyl)-2.}
`dimethyltetradecenalnide
`§—(4,6-dietl:oaty-S-pyriI'nidinyI)-2-
`methyltetradeeanamide
`(g)-§—(4,6-dimethory-5-pyfimidinyl}
`Lzdimcthyl-1 leicosensmlde
`2—metl1yl-y_—[l-(2-pyridinyl)el;hyl]-
`dodecauemidc
`1~=th¥1-H-[1-(1-PYl'idin¥1)ethy1]-
`dodecauamidc
`1-nr0w1-E—i1-(2-pyridinrl)ethrll-
`dodecanamide
`a-deci'1-!~[I-(1-r=vridinyl)ethyll-
`benznenepropanamidc
`1'm3|’-h¥l‘H‘“[1-(3-PY'~‘idi=IY1}91hYll-
`tetradeeanamide
`3-¢1|1l'1'E*‘[1'(3'P¥I'5d i-11¥1)=thf1l-
`tetradecanamide
`1-merhyl-§—[l-{2-pyr'ldiny1)ethy1]-
`hexadecanamidc
`0.7
`2.1<iimet.hyl-lll—[1-(2-pyridi.nyl)ethyl]~
`hexadecanarnide
`
`
`0.26
`
`1.3
`
`0.5
`
`0.‘l'E-
`
`3.3
`
`1.6
`
`3.?
`
`10
`
`0.8
`
`0.‘!
`
`[.4
`
`In Vivo Tests
`
`40
`
`45
`
`50
`
`55
`
`In the cholestrol-fed rabbit test, male, New Zealand
`.
`white rabbits weighing approximately 1 kg were fed a
`normal diet 40 g per day of rabbit chow (Purina NO.
`5321, Ralston Purina Co., 711 West Fuesser Road, Mas-
`coutah, 111., 62224», USA). After six days on this diet, the
`rabbits were fed 50 g per day for three days of a chol-
`sterol-enriched diet consisting of one part of a choles-
`terol-containing
`chow (Purina
`Catalog
`No.
`84-1206WLI, 0.25% cholesterol) and two parts of nor-
`mal chow. Next, the rabbits were fed 60 g per day for
`four days of a cholsterol-enriched diet consisting of two
`parts of a cholesterol-containing chow (Purina Catalog
`No. 84l206WLI, 0.25% cholesterol) and one part of
`normal chow.
`After this meal adaptation and cholesterol loading
`period, the test compounds of this invention were ad-
`ministered to the test animals in oral doses of 50 mg/kg
`of body weight thirty minutes prior to each meal for
`seven days. Control animals were administered vehicle
`only.
`The animals were sacrificed three hours after their
`last meal in the postabsorptive state. Serum cholesterol
`levels were determined for each animal, and the data
`appear in Table 2 expressed as percent change in serum
`cholesterol level compared to control.
`In the cholesterol-fed rat test, male, Sprague-Dawley
`rats (approximately 200 g body weight) were randomly
`divided into groups and provided ad libitum a regular
`rat chow diet (Purina No. 5002) supplemented with
`5.5% peanut oil, 1.5% cholesterol and 0.3% cholic acid,
`with or without drug admixed at the indicated levels
`(w/w). After one week, the animals (nonfasted) were
`etherized and blood was tak from the heart into
`EDTA (0.l4% final concentration) to measure total
`65 cholesterol using the Abbott VP Analyzer.
`The results of in vivo testing of representative com-
`pounds of the present invention are presented in Table
`2.
`
`5
`propriate alkyl halide. A second ct-alkyl substituent is
`attached by repeating this procedure.
`Details for the reaction conditions for preparing the
`tr-lithio carbanion salt of acids or esters, and for the
`conversion of these salts to a-alkyl-substituted acids or
`esters is found in P. Cneger, Org. Syn., Vol. 50, pp 58 if...
`John Wiley & Sons, New York, 1970.
`The substituted henzeneamine and substituted phe-
`nylmethylamine starting materials are prepared by
`methods well known in the art.
`The substituted pyrimidin-5-ylamines are prepared
`from the mono-, di-. or trichloropyrimidines by first
`nitrating the chloropyriinidines to produce the chlori-
`nated 5-nitropyrimidines. The chlorine substituents are
`then replaced by alkoxy substituents by heating the
`nitrochloropyrirnidines with the sodium salt of the de-
`sired alcohol in the same alcohol as solvent under reflux
`conditions. Following conversion of the ch]oro-5-
`nitropyrhnidines
`to the
`corresponding alkoxy-5-
`nitropyrimidines, the nitro group is reduced to an amine
`function in the conventional manner as, for example, by
`catalytic hydrogenation.
`As shown by the data presented below in Table 1 the
`compounds of the present invention are potent inhibi-
`tors of the enzyme acyl-CoA: cholesterol acyltra.nsl"er-
`ase (ACAT), and are thus eifective in inhibiting the
`esterification and transport of cholesterol across the
`intestinal cell wall.
`The ability of representative compounds of the pres-
`ent invention to inhibit ACAT was measured using an
`in vitro test more fully described in Field, F. J. and
`Salone, R. G., Biochemica et Biophysica 712: 557-570
`(1982). The test assesses the ability of a compound to
`inhibit the acylation of cholesterol by oleic acid by
`measuring the amount of radio-labeled cholesterol ole-
`ate formed from radio-labeled oleic acid in a tissue prep-
`aration containing rabbit intestinal microsomes.
`The data appear in Table 1 where they are expressed
`as [C59 values; i.e. the concentration of test compound
`required to inhibit cholesterol esterification by 50%.
`TABLE 1
`
`
`
`Compound
`lfl—[2,6-dime£hylphenyl}-2.1-di.metlryl-
`dodecsnsmide
`fl—(2¢tho:ry-E»methylphenyl)-2.2-
`dirnethyldodecansmide
`@)-fl—(‘2-methorry-6-melhylpheny])-2,2-
`dimethyl-1 1-eicosenamide
`2,2-dimethyl-§—(2,4,6-t1'imctl1oxy-
`phenyl)dodecan.emide
`2-met.hyl-I;T—(1.4.6-t.rimethoxyphcnyl}-
`tetradecanamide
`2-ethyl-§—(2,4,d-trimethoxyrphenyl}
`bctradccanamide
`2.2-di.ruethyl~E—(2,4.6-t.rimethoxy-
`phenyl)tetradecanaru.ide
`2-methyl-fi_—(2.4.6-trimethoxyphenyl}
`hexadecanamide
`2.2-di:nethy]-Ij—{2,4.ti-n-inretho:ty-
`phenyl)hexadecanamitle
`2.2-dirnethy]-§—(2.4.I5-t.rim:rho:ry-
`phenyboctademnamide
`l-decyl-1fl—(2.4.f>nimethcxypherryl)-
`cyclopentanccarbottatnide
`(§}-!.-methyl-fi—(2.4.6-trimethoxy-
`phenyl)-9-octadecensmide
`(Z)-2.2-dimethyl-§—(2,4.6-tri.methoJry-
`phenyl)-9-octsdeeenamide
`(§)-Z.2»dimc‘lh1|r]-fl-—(2,4.6—l1imethony-
`phenyl}-ll-eicosenamide
`E—(4.6-dimetlroxy-5-pyrimidiny1)-2.2-
`dimethyldodecanamide
`
`[C50
`(Micromolar)
`1.3
`
`0.23
`
`0.63
`
`0.042
`
`0.13
`
`0.05
`
`0.063
`
`0.031
`
`0.0-I4
`
`0.03‘.-‘
`
`0.007
`
`0.034
`
`0.0-14
`
`0.11
`
`0.23
`
`4ofl1
`
`PENN EX. 2195
`
`CFAD V. UPENN
`lPR20l5-01836
`
`

`
`7
`
`TABLE 2
`Percent
`Reduction in Cholesterol
`
`Compound
`Rabbit
`Rat
`2.2-Dimethyl-1~l_—(2.4,IS-tri-
`— 6?
`-54
`methoxyplienyl)dodecan.I.mide
`2-metby1-Ll—(2.4.6-I:rimethony-
`pheny1)taradeca.na.Inide
`§—{4,IS-di1nethoxy-5-pyrim-
`idinyl)-2-methyltetradecanamide
`l-Decyl-fl—(2,4,6-trimetboxy
`plJcny])cyclop¢m1anecarboxamide
`Li_—(4-.6-d.imethoxy-5-pyrimidi.nyl)-
`2-metizyltetradecanamide
`
`4,716,175
`
`ministered to the patient at dosage levels of from 500 to
`2000 mg per day. For a normal human adult of approxi-
`mately 70 kg of body weight, this translates into a dos-
`age of from '3’ to 30 mg/kg of body weight per day. The
`specific dosages employed, however, may be varied
`depending upon the requirements of the patient, the
`severity of the condition being treated, and the activity
`ofthe compound being employed. The determination of
`optimum dosages for a particular situation is within the
`skill of the art.
`
`The following preparative examples are provided to
`enable one skilled in the art to practice the invention,
`and are illustrative thereof. They are not to be read as
`limiting the scope of the invention as it is defined by the
`appended claims.
`
`Representative Example of the Preparation of an
`ct,ct-Dialkylalkanoic Acid
`
`Preparation of 2,2-Dimethyloctadecanoic Acid
`
`Diisopropyl amine (20.6 ml, 28.6 g, 0.283 mol) was
`dissolved in 250 ml of dry tetrahydrofuran. To this
`mixture was added 13.6 g (0.283 mol) of 50% sodium
`hydride. Isobutyric acid (26.2 ml, 24.9 g, 0.233 mol) was
`added dropwise with stirring and the temperature was
`allowed to rise. After addition of the acid was complete,
`the mixture was heated under reflux for an additional 20
`minutes. The mixture was then cooled to 0“ C. and 118
`ml (0.283 mol) of 2.4 M n-butyllithium was slowly
`added while maintaining the temperature below 5° C.
`When addition was complete. the mixtitrc was stirred at
`ice-bath temperature for l 5 minutes and then allowed to
`warm to room temperature and stirred for an additional
`two hours.
`
`The mixture was cooled to 0° C and 99.7 g (0.283
`mol) of 1-iodohexadecane were added dropwise. The
`resulting mixture was stirred at ice bath temperature for
`one hour, allowed to warm to room temperature and
`stirred at room temperature overnight.
`The mixture was again cooled to 0' C. and 400 ml of
`water was added with cooling. The aqueous layer was
`extracted with diethyl ether and the combined organic
`layers were dried and evaporated to yield a heavy gum.
`This material was taken up in hot water, the solution
`was made strongly acid with concentrated hydrochlo-
`ric acid. This mixture was extracted with diethyl ether,
`the ether layer separated, washed with brine, dried, and
`evaporated to yield 92.1 g of 2,2-dimethyloctadecanoic
`acid, mp 50-53“ C.
`
`Representative Example of the Preparation of an
`ct-Alkylalkanoic Acid
`(Alternative Method)
`
`5
`
`I5
`
`20
`
`30
`
`35
`
`50
`
`55
`
`Preparation of 2-Methylhexadecanoic Acid
`
`Sodium metal (12.06 g, 0.52 mol) was dissolved in 400
`ml of absolute ethanol. 2-Methyl-[,3-ptopanedioic acid,
`diethyl ester (95.3 g (0.55 mol) was added dropwise to
`the sodium ethoxide solution with stirring. When the
`addition was complete, the mixture was heated under
`reflux for 15 minutes.
`1-Bromotetradecane (133.65 g, 0.5 mol) was added
`dropwise with sfirring the the above mixture, and the
`resulting mixture was stirred and heated under reflux
`ovemight.
`After this time, the mixture was cooled, neutralized
`with acetic acid, and concentrated under vacuum to
`half its original volume. This residue was diluted with
`
`65
`
`Sofll
`
`PENN EX. 2195
`
`CFAD V. UPENN
`lPR20l5-01836
`
`—-45
`
`-29
`
`—-52
`
`-29
`
`— S2
`
`— 14$
`
`For preparing phannaoeutical compositions from the
`compounds of this invention, inert, pharmaceutically
`acceptable carriers can be either solid or liquid. Solid
`form preparations include powders, tablets, dispersihle
`granules, capsules, and cachets.
`A solid carrier can be one or more substances which
`may also act as diluents, flavoring agents, solubilizers,
`lubricants, suspending agents, binders, or tablet disinte-
`grating agents; it can also be an encapsulating material.
`In powders, the carrier is a finely divided solid which
`is in a mixture with the finely divided active compo-
`nent. In tablets, the active compound is mixed with the
`carrier having the necessary binding properties in suit-
`able proportions and compacted in the shape and size
`desired.
`Powders and tablets preferably contain between
`about 5 to about 70% by weight of the active ingredi-
`-rent. Suitable carriers are magnesium carbonate, magne-
`sium stearate, talc, lactose, sugar, pectin, dextrin, starch,
`tragacanth, methyl cellulose, sodium carboxymethyl
`cellulose, a low-melting wax, cocoa butter, and the like.
`The term “preparation” is intended to include the
`formulation of the active compound with encapsulating
`.-material as a carrier providing a capsule in which the
`active component (with or without other carriers) is
`.surrounded by a carrier, which is thus in association
`-with it. In a similar manner, cachets are also included.
`Tablets, powders. cachets, and capsules can be used
`as solid dosage forms suitable for oral administration.
`Liquid form preparations include solutions suitable
`for oral administration, or suspensions and emulsions
`suitable for oral administration. Aqueous solutions for
`oral administration can be prepared by dissolving the
`active compound in water and adding suitable flavor-
`ants, coloring agents, stabilizers, and thickening agents
`as desired. Aqueous suspensions for oral use can be
`made by dispersing the finely divided active component
`in water together with a viscous material such as natural
`or synthetic gums, resins, methyl cellulose, sodium car-
`boxymethyl cellulose, and other suspding agents
`known to the pharmaceutical formulation art.
`Preferably, the pharmaceutical preparation in is unit
`dosage form. In such form, the preparation is divided
`into unit doses containing appropriate quantities of the
`active component. The unit dosage form can be a pack-
`aged preparation, the package containing discrete quan-
`tities of the preparation, for example, paclteted tablets,
`capsules, and powders in vials or ampoul. The unit
`dosage form can also he a capsule, cachet, or tablet
`itself, or it can be the appropriate number of any of
`these packaged forms.
`In therapeutic use as agents for the inhibition of intes-
`tinal absorption of cholesterol, the compounds utilized
`in the pharmaceutical method of this invention are ad-
`
`

`
`4,716,175
`
`10
`TABLE 3-continued
`
`E*"“"'P'° C°‘"‘P°“""
`M-F ('9)
`10
`2-1'D=fl1lv'l-1£—{2.4.6-trimethox)'-
`109-1 11
`Ph=n¥1)”="9d==W31¥'5d=
`’f=’1‘)'3‘“""-‘“"?°"*°*’*
`‘,’_,°j,‘{mf,,",°“,.N°““‘”"_,,.,,,f
`trintelhoxyphenyntetradecartamide
`2.mou:y1-LI—(2.4.eh-houhoxy-
`phenyllhexademamidc
`,
`2*_7"‘“$“h3"’§‘U""5‘
`%::§';,}_':,'E12;1_:‘:_d'°°"‘m“°
`tfimmmwhgnylhcgdummfide
`1-ooq1—_1g—{2,4.sm'mea.oxy—
`tihenvlmclohntanccarboxamide
`""°“3"'§‘(2*“-""“"""“'°"Y'_
`Ph°"3'D°3'°]°P°”m°°”b°”“"d°
`{Z)o2-1nethyl-N—{2,4,6-
`,,im,,h°,yph;,m_,mmdwmmjdc
`@-z.2-dimethv1-§-—{1.4,6-
`uimeu1oxypheo1yg—9.o;:1;idooohom5oe
`2,2-dirnet
`-
`.
`,5-
`?mou1oxypht:;y1T.E(-eioooensmide
`§—(4,&dimethoxy-5-pyrimidinyl}
`2.2<iimetJIyldodecanam.ide
`§—(4,o-aimothoxy-2-phony]-5
`pyfimidinrl)-2.2-dlmethrldodecannmidc
`§;*,f.;‘,‘,‘,°‘,$‘,I,",‘,’,’,‘,’§;f,',‘,’,,";""“"“"1"
`g-—(4,s-dsneshoxy-s-pyrim1d1hy1y
`2-ethrltetljadecannmide
`:—“‘(f'“'5';"1‘l""""°"3"’5'l’3"i’_"i"-""‘3'D'
`rgfiffigfifigfafigfigyl}
`‘§m,u,,,m,_,,,dw,,,,:,,‘d,
`taooyt-g—(4,s.oimou:oxy.
`Prrimidin-5-rbcrciopenbanecarboxamidc
`ll-eioosenamide
`§mm,_E_[H,_PymY,}
`gt]-,y]]dodacmnm|'de
`2-ethyl-fl—[l-(2-pyridi.nyl}-
`=1h¥1ldDd=¢=-Ilamide
`_
`_
`f'_;}1:"]’1‘fl"O"‘i5‘[1'{?ljl1""“"""")'
`,H,g,,_N_[,_(2_P;n»d,ny1>
`,,h,,1]1,,,,',,_.,,,,,,.,,,.,,_,,,,,.,.,
`2-methyl-g—{1—(2.om‘a1oy1y
`ethrlltnradecammidt-_
`1-="1¥‘-E-I143-P=rnd'“=">
`1}
`
`“
`,,
`
`13
`
`'4
`,5
`
`15
`
`17
`18
`
`19
`20
`
`21
`
`22
`
`2’
`24
`
`25
`15
`
`27
`
`R
`,,
`
`so
`
`3'
`32
`
`33
`
`93*”
`M,
`
`109-111
`
`5345
`6340
`
`$19.90
`
`734“
`Wax
`
`Wax
`Wax
`
`100-101
`
`93-99
`
`“°‘”°
`115-115
`
`9”‘
`m6_m.,
`
`101-103
`
`Wm
`W,
`
`12:14
`
`"'73
`5830
`
`53-71
`
`3‘
`3,
`
`54-35
`SW
`
`20
`
`1’-5
`
`35
`
`9
`water and the aqueous phase was separated and ex-
`tracted twice with diethyl ether. The organic layers
`were combined, washed with water, dried over a.nhy-
`drous magnesium sulfate, and evaporated to yield an oil.
`This oil was mixed with 112 g (1.7 mo1)of85% potas- 5
`sium hydroxide in 900 ml of 95% ethanol, and the re—
`suiting mixture heated to reflux. After about one-half
`hour, the reaction became quite vigorous. The mixture
`was stirred under reluit overnight, cooled to room tem-
`perature, and made strongly acidic with concentrated 10
`hydrochloric acid. The mixture was cooled and filtered.
`The solid was taken up in diethyl ether, dried, and evap-
`orated under vacuum to yield 2-methy1-2-ter.radecylma-
`Ionic acid, mp 83-85‘ C.
`The solid was heated with stirring to 165' C., wherc- 15
`upon evolution of CO: began. The temperature rose
`rapidly to 190' C. with rapid evolution of CO;. The
`.
`.
`.
`solid was then heated for an additional 1/2 hour at
`185-190“ C. to yield 81.3 g of 2-methyl-hexadeeanoic
`acid. mp 4446' C.
`Representative Example of the Preparation ofan
`Amide
`_
`Preparation of
`N-2,4,6-trimethoxyphenyl)-2-methylhexadecanamide
`2-Methylhexadecanoic acid (27.0 g 0.1 mol) was
`mixed with 100 ml of thionyl chloride and the resulting
`mixture was stirred and heated under reflux for eight
`hours and then stirred at room temperature overnight. 30
`The mixture was concentrated under vacuum, diethyl
`ether was added and the mixture again concentrated
`under vacuum. The residue was distilled to yield 25.8 g
`of 2-methylhexadecanoyl chloride, bp 120-125‘ C. at
`0'25 mm Hg’
`2,4,6-Trtmethoxyphenylamme hydrochloride (ass g,
`0.03 11101) and 3.3 ml (6.06 3, 0.06 mol) of tnelhylamtne
`were dissolved in 100 ml of tetrahydrofuran. To this
`mixture was slowly added, with stirring, 8.65 g of 2-
`methylhexa-decanoyl chloride. The resulting mixture 4.0
`was stirred at room temperature overnight, filtered, and
`the filtrate concentrated under vacuum. Water was
`added to the residue, the resulting solid collected by
`filtration, and recrystallized from isopropyl ether to
`yield 12.0 g of N-2,4,6- trimethoJtypher1yl)-2-methyl- 45
`hexadecanamide, mp 109-111" c.
`Employing the general methods detailed above, the
`following compounds in accordance with the present
`invention were prepared.
`TABLE 3
`Example Compound
`1
`E_{2’6_dimethylPhcnYn_m_
`dimethyldodecanamide
`2
`1'_'I—{2.5-dlethylphenyl)-2,‘}
`ditnethyldodecanamide
`E—[1.5-bic(l-methylcthyl}
`phenylI-2.,2-dimet.h3rl-
`dodecanamitle
`§—{2-o:hoxyo-meun-1pheny1)-
`1,2-dimetltyldodecanemide
`1-met.hyl-fi—[2,6-his(]-|netltyl-
`ethyl)pl1enyl1t_etradecanamide
`‘%3;%_T_'&c5'dt.f:e'£YlPhE“¥"2'
`@_N_{2’5_dmh'ylum‘Phy°1)_
`2,2-d_i.methyl-9-octadeceuamide
`{§l-E—{2-me_tho=o~6-methrl-
`”.h'"3")‘2'3{'£“n°‘h"1'l]'
`1’2.d|_m=thy[_}_,[_(2,4,5_
`t1-irnethoxyrpheng.-l)dodecanamide
`
`etl1y1]he:ta:leca.u.Imide
`Y
`as
`2,2-o1oiothy1.§_[1.(2.pyn‘aihy1)-
`32-33
`
`ethvllhexadecarlamide
`50 W chi
`_
`I“:
`3
`l.Acompo1md havmg the formula:
`R,
`0
`1
`/w
`A—C—C
`l
`R2
`
`3
`
`4
`s
`
`5
`-,
`
`8
`
`g
`
`M.p. cc.)
`S344
`‘I-'0-'32
`
`119-120
`
`so-52
`94-96
`
`Wax
`W“
`
`Wax
`
`5940
`
`55
`
`-
`
`\
`
`N—1’l
`,',
`
`60
`
`55
`
`wherein A is an unbranched hydrocarbon group con-
`taioing from one to twenty carbon atoms and which
`may contain from one to three carbon-carbon double
`bonds;
`R:
`is hydrogen, alkyl of from one to four carbon
`atoms or phenyhnethyl;
`R2 is alkyl of from one to four carbon atoms or phe-
`nylmethyl; or
`
`6 0”‘
`
`PENN Ex. 2195
`CFAD V. UPENN
`lPR20l5-01836
`
`

`
`11
`R; and R2 when taken together with the carbon atom
`to which they are attached form a saturated carbo-
`cyclic ring of from three to seven carbon atoms;
`B is selected from
`
`I
`(CH2);
`
`Rs
`
`R4
`
`R3
`
`('21:;
`N
`and—CH
`
`where n is zero or one,
`R3, R4, and R5 are independently selected from
`tri-
`hydrogen,
`fluorine,
`chlorine.
`bromine,
`fluromethyl, alkyl of from one to four carbon
`atoms, and alkoxy of from one to four carbon
`atoms;
`2. A compound inaccordance with claim I wherein B
`
`is
`
`3. A compound in accordance with claim 1 wherein B
`
`is
`
`4,716,175
`
`12
`
`(Z)-N-
`-{2,6-diethylphenyl-2-methy1-9-octadecenamide;
`(Z)-N-(2,6-diethylphenyl)-2,2-dimethyl-%
`octadecenamide;
`(Z)-N-(2-methoxy-6-methylphenyl)-2,2-dimethyl-1 1-
`eicosenamide;
`2,2-dimethyl-N-(2.4,6-trimethoxyphenyl)dodecana—
`mide;
`2-methyl-N—(2,4,6-trimethoxyphcny1)tetradccana-
`mide;
`2~ethyl-N~»[2,4,6-trimethoxyphenyl)tetradecanamide;
`2,2-ditnethyl-N—(-Z4,6-t1'imethoxypheny1)tet-
`radecanamide;
`2-methyl-N-(2.4-,6-trimethoxypheny1)hexadecana-
`mide;
`2,2-dimethyl-N1(2,4,6-trimethoxyphenyl)hexadecana—
`mide;
`2,2-dimethyl-N-(2.4,6-t1'imethoxypl'1enyl)octadecana-
`wide;
`1-decyl-N-{2,4,6-trimethoxypheny1)cyclobutanecar-
`boxamide;
`I-decyl-N-(2,4,6-trimethoxyphenyl)cyc1opentanecar-
`boxamicle;
`(Z)-2-methyl-N-(2,4»,6-trimethoxyphenyl)-9-
`octadecenamide;
`(Z)-2,2-dimethyl-N-(2,4,6-trimethoxyphenyl}
`octadecenamide; and
`(Z)-2,2-dimethyl-N-(2,4»,6-trimethoxyphenyl)-I 1-
`eicoseuamide.
`5. A compound in accordance with claim 3 selected
`from the group consisting of
`2-methy1—N-[1-(2-pyridinyhethyldodecanamide;
`2-ethyl-N-I1-(2-pyridinyl)ethyl}doclecanamide;
`2-propyl-N-[l{2-pyridinyl)ethyl]dodecanamide;
`ct-decyl~N-[l-(2-pyridinyl)ethyl]benzenepropana-
`mide;
`2-methyl-N-[1-(2-pyridinyl)ethyl}tetradecan.an1ide;
`2-ethyl-N-[1-(2-pyridinyl)ethy1]te'tradecanan1ide;
`2-methyl-N-I1-(2-pyridinyl)ethyl]h.exadecanamide;
`and
`2,2-dimethyl-N-[1-(2-pyridinyl]ethyl}hexadecana—
`mide.
`
`10
`
`I5
`
`25
`
`30
`
`35
`
`4. A compound in accordance with claim 2 selected
`from the group consisting of
`N-(2,6-dimethylpheny1)-2.2-dimethyldodecanamide;
`N-(2,6-diethylphenyl)-2,2-dimethyldodecanamide;
`N-[2,6-bis(1-1nethyletl1yl)phenyl]-2,2-dimethyl-
`dodecanamide:
`
`N-(2-ethoxy-6-methylphenyl)-2,2-dimethyldodecana»
`mide;
`2-methyl-N-[2,6-bis(1-methylethyl)pheny1]tet-
`radecanamide;
`
`6. A pliarmaceutical composition useful for inhibiting
`the intestinal absorption of cholesterol comprising an
`45 ACAT-inhibitory effective amount of a compound as
`defined by claim 1 in combination with a pharmaceuti-
`cally acceptable carrier.
`1-‘. A method of inhibiting intestinal absorption of
`cholesterol comprising administering to a patient an
`ACAT-inhibitory effective amount of a compound as
`defined by claim 1 in combination with a pharmaceuti-
`cally acceptable carrier.
`F
`I
`I
`II
`I
`
`50
`
`7ofl1
`
`PENN EX. 2195
`
`CFAD V. UPENN
`lPR20l5-01836
`
`

`
`UNITED