[11] Patent Number:
`
`4,761,419
`
`[45] Date of Patent:
`
`Aug. 2, 1988
`
`3/1931 Oka eta]. .......................... .; 424/279
`4,255,444
`4/1931 Mistuietal.
`.. 424/279
`4,262,013
`3/1933 Willard etal..
`424/279
`4,375,475
`4,613,610 9/1986 Wareing ....... ..
`.. 514/406
`4,647,576
`3/1987 Hoefle etal.
`.514/422
`4,668,794
`5/1987 Wareing
`.. 543/342
`4,681,893
`7/1987 Roth .................................. .. 514/422
`
`
`
`OTHER PUBLICATIONS
`
`PCT International Application No. PCT/EP83/00308
`Pub. No. W0 84/02131.
`
`Primary Exam1'ner——Mary C. Lee
`Assistant Examiner—-J. Richter
`Attorney, Agent, or Fz'rm———Jerry F. Janssen
`
`[57]
`
`ABSTRACT
`
`trans-6-[[(substituted)quinolinyl]ethyl]-and
`Certain
`ethenyl]tetrahydro-4-hydroxypyran-2-ones and the cor-
`responding dihydroxy ring-opened acids derived there-
`from are potent inhibitors of the enzyme 3-hydroxy-3-
`methylglutaryl-coenzy'me A reductase (HMG-CoA
`reductase) and are useful as hypocholesterolemic and
`hypolipidemic agents.
`
`United States Patent
`Picard et al.
`
`[19]
`
`[54]
`
`6-(((SUBSTITUTED)QUINOLINYL)ETHYL)-
`AND
`ETHENYDTETRAHYDRO-4-HYDROXYPY-
`RAN-2-ONE INHIBITORS OF
`CHOLESTEROL BIOSYNTHESIS
`
`[75]
`
`Inventors:
`
`Joseph A. Picard; Bruce D. Roth,
`both of Ann Arbor; Drago R.
`Sliskovic, Ypsilanti, all of Mich.
`
`[73]
`
`[21]
`
`[22]
`
`[51]
`
`[52]
`
`[58]
`
`[56]
`
`Assignee: Warner-Lambert Company, Morris
`Plains, N.J.
`
`Appl. No.: 129,516
`
`Filed:
`
`Dec. 7, 1987
`
`Int. Cl.4 ..................... A61K 31/34; A61K 31/47;
`CO7D 217/12; C07D 217/14
`U.S. Cl. .................................. .. 514/311; 514/256;
`514/314; 546/ 167; 546/171; 546/173; 546/174;
`546/175; 544/333
`Field of Search ............. .. 546/167, 171, 173, 174,
`546/175; 544/333; 514/311, 314, 256
`References Cited
`
`U.S. PATENT DOCUMENTS
`
`4,198,425 4/1980 Mistui et al.
`
`...................... .. 424/279
`
`15 Claims, No Drawings
`
`Mylan Exhibit 1021, Page 1
`
`Mylan Exhibit 1021, Page 1
`
`

`
`1
`
`4,761,419
`
`6-(((SUBSTITUTED)QUINOLINYL)ETHYL)-AND
`ETHENYL)TETRAHYDRO-4-HYDROXYPYRAN-
`2-ONE INHIBITORS OF CHOLESTEROL
`BIOSYNTHESIS
`
`BACKGROUND OF THE INVENTION
`
`The present invention is related to compounds and
`pharmaceutical compositions useful as hypocholestero-
`lemic and hypolipidemic agents. More particularly, this
`invention
`concerns
`certain
`trans-6-[[(substituted)-
`quino1inyl]ethy1]- and ethenyl]tetrahydro-4-hydroxypy-
`ran-2-ones and the corresponding dihydroxy ring-
`opened acids which are potent inhibitors of the enzyme
`3-hydroxy-3-methylglutaryl-coenzyme A reductase
`(HMG-CoA reductase), pharmaceutical compositions
`containing such compounds, and a method of lowering
`blood serum cholesterol levels employing such pharma-
`ceutical compositions.
`High levels of blood cholesterol and blood lipids are
`conditions which are involved in the onset of arterio-
`sclerosis. It is well known that inhibitors of HMG-CoA
`reductase are effective in lowering the level of blood
`plasma cholesterol, especially low density lipoprotein
`cholesterol (LDL-C), in man (cf. M. S. Brown and J. L.
`Goldstein, New England Journal of Medicine (1981),
`305, No. 9, 515-517). It has now been established that
`lowering LDL-C levels affords protection from coro-
`nary heart disease (of. Journal of the American Medical
`Association (1984) 251, No. 3, 351-374).
`Moreover,
`it is known that certain derivatives of
`mevalonic acid (3,5-dihydroxy-3-methylpentanoic acid)
`and the corresponding ring-closed lactone form,
`mevalonolactone, inhibit the biosynthesis of cholesterol
`(cf. F. M. Singer et al, Proc. Soc. Exper. Biol. Med.
`(1959), 102, 270) and F. H. Hulcher, Arch. Biochem.
`Biophys.. 30 (1971), 146, 22.
`U.S. Pat. Nos. 3,983,140; 4,049,495 and 4,137,322
`disclose the ferrnentative production of a natural prod-
`uct, now called compactin, having an inhibitory effect
`on cholesterol biosynthesis. Compactin has been shown
`to have a complex structure which includes
`a
`mevalonolactone moiety (Brown et al, J. Chem Soc.
`Perkin L (1976), 1165.
`U.S. Pat. No. 4,255,444 to Oka et al, discloses several
`synthetic derivatives of mevalonolactone having an-
`tilipidemic activity.
`U.S. Pat. Nos. 4,198,425 and 4,262,013 to Mitsue et al,
`disclose aralkyl derivatives of mevalonolactone which
`are useful in the treatment of hyperlipidemia.
`U.S. Pat. No. 4,375,475 to Willard et al, discloses
`certain substituted 4-hydroxytetrahydropyran-2-ones
`which, in the 4(R)-trans stereoisomeric form, are inhibi-
`tors of cholesterol biosynthesis.
`U.S. Pat. No. 4,647,576 to Hoefle, et al, discloses
`certain
`trans-6-[2-[(substituted)-pyrrol-1-yl]-]alkyltet-
`rahydro-4-hydroxypyran-2-ones and the corresponding
`lactone ring-opened acids as inhibitors of cholesterol
`biosynthesis.
`U.S. Pat. No. 4,681,893 to Roth discloses certain
`trans-6-[[(2-, (3-, or (-carboxamido-substituted)pyrrol-1-
`yl]alkyl- or alkenyl]-tetrahydro4-hydroxypyran-2-one
`inhibitors of cholesterol biosynthesis.
`
`SUMMARY OF THE INVENTION
`
`In accordance with the present invention, there are
`provided certain trans-6-[[2-(substituted)quinolinyl]-
`ethyl- or
`ethenyl]tetrahydro-4-hydroxypyran-2-ones
`
`10
`
`15
`
`20
`
`25
`
`30
`
`35
`
`45
`
`50
`
`55
`
`60
`
`65
`
`2
`and the corresponding ring-opened hydroxy-acids
`which are potent inhibitors of cholesterol biosynthesis
`by virtue of their ability to inhibit
`the enzyme 3-
`hydroxy-3methylglutaryl
`coenzyme A reductase
`(HMG-CoA reductase).
`In particular,
`in its broadest chemical compound
`aspect, the present invention provides compounds of
`structural Formula I
`
`R3
`
`R1
`
`I
`
`R4
`
`Rs
`
`X-A
`
`N
`
`R5
`
`R2
`
`wherein A is
`
`Hot H
`
`I III
`
`H
`
`§0
`
`O
`
`and X is ——CH2CH2— or _—CH=CH— (preferably in
`the trans configuration).
`R1 and R2 are independently hydrogen; alkyl of from
`one to six carbons; trifluoromethyl; cyclopropyl; cyclo-
`hexyl; cyclohexylmethyl; phenyl; phenyl substituted
`with fluorine, chlorine, bromine, hydroxy,
`trifluoro-
`methyl, alkyl of from one to four carbon atoms, or
`alkoxy of from one to four carbon atoms; phenylmethyl;
`phenylmethyl substituted with fluorine, chlorine, bro-
`mine, hydroxy, trifluoromethyl, alkyl of from one to
`four carbon atoms, or alkoxy of from one to four carbon
`atoms; 2-, 3-, or 4-pyridinyl; or 2-, -, or 5-pyrimidinyl;
`provided that when X is in the 2-position, R1 is hydro-
`gen and is attached in the 4-position.
`R3, R4, R5, and R5 are independently selected from
`hydrogen; alkyl of from one to six carbon atoms; trifluo-
`romethyl; cyclopropyl;
`fluorine; chlorine; bromine;
`hydroxy; alkoxy of from one to four carbon atoms;
`cyano; nitro; amino; acetylamino; aminomethyl; phenyl;
`phenyl substituted with fluorine, chlorine, bromine,
`hydroxy,
`trifluoromethyl, alkyl of from one to four
`carbon atoms, or alkoxy of from one to four carbon
`atoms; phenylmethyl; or phenylmethyl substituted with
`fluorine, chlorine, bromine, hydroxy, trifluoromethyl,
`or alkyl of from one to four carbon atoms.
`Also contemplated as falling within this aspect of the
`invention are the corresponding dihydroxy-acid com-
`pounds of Formula II corresponding to the opened
`form of the lactone ring of compounds of Formula I
`
`\‘
`s‘ll
`
`where X, R1, R2, R3, R4, R5, and R5 are as defined
`above,
`and the pharmaceutically acceptable salts
`
`Mylan Exhibit 1021, Page 2
`
`Mylan Exhibit 1021, Page 2
`
`

`
`4,761,419
`
`3
`thereof, all of the compounds being in the trans race-
`mate of the tetrahydropyran moiety.
`In another aspect of the present invention, there is
`provided a method of preparing compounds of Formula
`I above by (a) first reacting a substituted [(quinolin-3-
`yl)ethyl- or etheny1]aldehyde compound of Formula III
`
`5
`
`4 111
`
`10
`
`is substituted at the 2-, 3-, or -position with the group
`
`HO
`
`Ho
`S\
`
`where are X, R1, R2, R3, R4, R5, and R5 as defined
`above, with the alkali metal salt of the dianion of ethyl
`acetoacetate to form a compound of structural Formula
`IV
`
`IV
`
`in’
`X—CH(OH)CH2CCH2COOC2H5
`
`R5
`
`R2
`
`where X, R1, R2, R3, R4, R5, and R5 are as defined
`above, then successively (b) reducing Compound IV
`with a trialkylborane and sodium borohydride and (c)
`oxidizing with alkaline hydrogen peroxide to produce
`an ester compound of Formula V
`
`R4
`
`Rs
`
`R3
`
`R1
`
`V
`
`\.H
`,1‘! H0
`H0
`\
`\
`X—C"—"CH2—-C—CH2COOC2H5
`
`N
`
`R5
`
`R2
`
`and finally (d) hydrolyzing and cyclizing, if desired, the
`ester compound of Formula V to a lactone compound
`of Formula I by heating in an inert solvent or, alterna-
`tively converting,
`if desired,
`the intermediate dihy-
`droxy acid thus formed to a pharmaceutically accept-
`able salt.
`
`the present invention provides
`In another aspect,
`pharmaceutical compositions, useful as hypolipidemic
`or hypocholesterolemic agents, comprising a hypolipi-
`demic or hypocholesterolemic affective amount of a
`compound in accordance with this invention as set forth
`above, in combination with a pharmaceutically accept-
`able carrier.
`
`In another aspect, the present invention provides a
`method of inhibiting cholesterol biosynthesis in a pa-
`tient in need of such treatment by administering a phar-
`maceutical composition in accordance with the present
`invention as defined above.
`
`DETAILED DESCRIPTION
`
`The compounds of the present invention form a class
`of substituted quinolines in which the quinoline moiety
`
`15
`
`20
`
`25
`
`30
`
`35
`
`45
`
`50
`
`55
`
`60
`
`65
`
`where X is as previously defined. Compounds in which
`X is attached at position 2 and groups other than hydro-
`gen are attached at position 4 of the quinolinyl moiety
`are difficult to synthesize, and are thus excluded from
`the scope of this invention.
`Preferred compounds of the invention are those in
`which the position of attachment of the ethyl- or ethe-
`nyl-lactone ring is at position 3 of the quinolinyl moiety.
`Preferred substituent groups for R1 and R2 are phenyl,
`substituted phenyl, and lower alkyl, most preferably
`isopropyl.
`As used throughout this specification and the ap-
`pended claims, the term “alkyl” denotes a branched or
`unbranched saturated hydrocarbon group derived by
`the removal of one hydrogen atom from an alkane. The
`term “lower alkyl” denotes alkyl of from one to four
`carbon atoms.
`
`The term “alkoxy” denotes an alkyl group, as just
`defined, attached to the parent molecular
`residue
`through an oxygen atom.
`Particularly preferred compounds of the present in-
`vention include the following:
`[4a,6B(E)]6-[2-[6-Chloro--
`-(4-fluorophenyl)-2-methyl-3-quinolinyl]ethenyl]tet-
`rahydro-4~hydroxy-2H-pyran-2-one.
`[4oL,6B(E)]6-[2-[6-Chloro-4-(-fluorophenyl)-2-(1-
`methylethyl)-3-quinolinyl]ethenyl]tetrahydro-4-
`hydroxy2H-pyran-2-one.
`[4a,6B(E)]6-[2-[2-[6-(-4Fluorophenyl)-2-(1-methyle-
`thyl)-3-quinolinyl]ethenyl]tetrahydro-4-hydroxy-2H-
`pyran-2-one.
`[R*,S"‘-(E)]-7-[6-Chloro-4-(4-fluorophenyl)-2-methyl-3-
`quinolinyl]-3,5-dihydroxy-6-heptenoic acid.
`[R*,S*
`—(E)]-7-[6-Chloro-4»(4-fluorophenyl)-2-(l-
`methylethyl)-3-quinolinyl]-3,5-dihydroxy-6—hep-
`tenoic acid.
`
`[R*,S*-(E)]-7-[4-(4-Fluorophenyl)-2-(l-methylethyl)-3-
`quinolinyl]-3,5-dihydroxy-6-heptenoic acid.
`Compounds of the present invention in which the
`ethyl- or ethenyl-lactone moiety is attached to position
`3 of the quinolinyl moiety are prepared by the general
`synthetic methods outlined in Reaction Sequence 1.
`Compounds of the present invention where the ethyl-
`or ethenyl-lactone moiety is attached to the 2- or 4-posi-
`tion of the quinolinyl moiety are prepared by the gen-
`eral synthetic methods outlined in Reaction Sequence 2.
`
`Mylan Exhibit 1021, Page 3
`
`Mylan Exhibit 1021, Page 3
`
`

`
`5
`
`4,761,419
`
`6
`
`Reacfion Seguencel
`
`/
`R1—C\
`
`0
`
`C1
`
`1
`
`+
`
`R6
`
`NH2
`
`R5
`2
`
`R3
`
`R4
`
`———-9
`
`R5
`
`R1
`
`0
`
`\ /
`C
`
`NH2
`
`R3
`
`R4
`3
`
`R3
`
`R1
`
`R4
`
`R5
`
`R6
`
`‘ CHZOH
`/
`
`N
`
`R2
`
`COOR
`
`CHO
`
`
`CHZOH
`
`COOR
`
`CI-I20!-I
`
`
`
`10
`
`11
`
`Mylanv Exhibit 1021, Page 4
`
`Mylan Exhibit 1021, Page 4
`
`

`
`4,761,419
`
`-continued
`Reaction Sequence 1
`
`COOEt
`
`l3a—l3b
`
`
`
`l2a— 1 2b
`
`Cl-{O
`
`
`
`R2
`
`R1
`
`\ I
`
`N
`
`R3
`
`R6
`
`
`
`14a—l4b
`
`Referring to Reaction Sequence 1, the acid chloride,
`1 having the desired substituent group R1,
`is reacted
`with the appropriately 2,3,4,5-substitued aniline, 2, in
`the presence of zinc chloride at a temperature of about
`200° C., and the resulting reaction mixture hydrolyzed
`with acetic and hydrochloric acids to produce the sub-
`stituted aclamine, 3.
`The acylamine, 3, is reacted with the desired B-l<eto-
`ester, 4, in the presence of p-toluenesulfonic acid, an the
`intermediate thus formed is dehydrated and cyclized to
`the substituted quinoline-3-carboxylic acid ester, 5, by
`heating in toluene under reflux.
`'
`The ester, 5, is reduced at -78° C. by the action of
`diisobutylaluminum hydride (“DIABL”) to yield the
`alcohol, 6. The alcohol, 6, is then oxidized to the corre-
`sponding aldehyde, 7, by the method of Swem (Swem,
`et al, J, Org. Chem, 43:2480 (1978) to yield the desired
`aldehyde, 7.
`Wittig reaction of the aldehyde, 7, with an ylide such
`as carbomethoxy triphenylphosphorane in methylene
`chloride at room temperature produces the unsaturated
`trans-ester, 8, in high yield. T ester, 8, is reduced to the
`allyl alcohol, 9, using a well-known procedure employ-
`ing two equivalents of diisobutyl aluminum hydride at
`-78° C.
`is reduced
`the unsaturated ester, 8,
`Alternatively,
`over Pd/C by the action of hydrogen to produce the
`saturated ester, 10, which is then reduced by the action
`of DIBAL to produce the corresponding alcohol which
`may then be carried forward in the sequence of steps to
`finally produce the product having the saturated ethyl
`bridge (X=ethylene in generic Formula I).
`The alcohols, 9 or 11, are reoxidized to the corre-
`sponding aldehydes, 12a or 12b, by Swem oxidation,
`followed by an aldol condensation with the sodium
`
`35
`
`40
`
`45
`
`50
`
`55
`
`60
`
`65
`
`in
`lithium dianion of ethyl acetoacetate at -78° C.
`tetrahydrofuran (See Kraus, et al,
`J. Org. Chem.,
`4822111 (1983) to form the 5-hydroxy-3-oxo-6-heptenoic
`esters 13a and 13b.
`The product of this condensation is then reduced in a
`sequence of steps in which it is first dissolved in a polar
`solvent such as tetrahydrofuran under a dry atmo-
`sphere. A small excess of triethylborane and catalytic
`amounts of 2,2-dimethylpropanoic acid are next added.
`The mixture is stirred at room temperature for a short
`period, after which it is cooled to a temperature prefera-
`bly between about —60° C. and —80° C. Dry methanol
`is added, followed by sodium borohydride. The mixture
`is kept at low temperature for 4-8 hours before treating
`it with hydrogen peroxide and ice water. The substi-
`tuted 3,5-dihydroxy-6-heptenoic acid ethyl esters, 14a
`and 14b, are isolated" having the preferred R*,S* and
`R*,R* configurations, respectively.
`The esters, 14a and 14b may be utilized as such in the
`pharmaceutical method of this invention, or may be
`converted, if desired, to the corresponding acid salt
`forms, such as the sodium salt, employing basic hydrol-
`sis by generally well-known methods. The free acids,
`produced by acidification of the sodium salts, can be
`dehydrated to the lactones, I by heating the acids in an
`inert solvent such as toluene with concomitant azeo-
`tropic removal of water.
`Referring to Reaction Sequence 2, the substituted
`isatin 15, is condensed by the Pfitzinger Reaction (see
`W. Pfitzinger, J. Prakt. Chem, [2]3:lO0 (1886); 38:582
`(1888)) with the oxime, 16, to produce the substituted
`quinoline-4-carboxylic acid, 17. Alternatively, the po-
`tassium salt of 17 (prepared by the treatment of acid, 17,
`with potassium hydroxide in
`
`Mylan Exhibit 1021, Page 5
`
`Mylan Exhibit 1021, Page 5
`
`

`
`9
`
`4,761,419
`
`Reaction Sequence 2
`
`=0
`
`+
`
`R2—CI-l2—C
`
`N-OH
`
`R1
`
`—%
`
`R4
`
`R5
`
`16
`
`"CO2
`
`R1 = H
`
`10
`
`R3
`
`COZH
`
`©
`N
`
`R5
`
`17
`
`R2
`
`CHO
`
`Haack
`é Vilsmeier-
`
`R4
`
`R5
`
`R3
`
`H
`
`R3
`
`COZR
`
`R2
`
`R4
`
`N
`
`H
`
`R5
`
`N
`
`R5
`
`19
`
`R5
`
`18
`
`R2
`
`R1
`
`R2
`
`R]
`
`R4
`
`R5
`
`R4
`
`R5
`
`R
`
`N
`1'.‘
`
`H
`
`N
`
`R3
`
`R5
`
`R3
`
`R5
`
`15
`
`20
`
`Steps similar
`to Sequence 1
`
`A
`
`Steps similar
`to Sequence 1
`
`OH
`\\“H
`
`0 R0
`
`R2
`
`R1
`
`
`
`I
`
`0
`
`¢
`
`R;
`
`H
`
`R4
`
`R5
`
`R6
`
`R2
`
`0 1,,” H O
`
`"/1
`
`N
`
`OH
`
`methanol) is esterified by heating a tetrahydrofuran
`solution of the salt of 17 with ethyl iodide at a tempera-
`ture of about 70° C. The resulting ethyl ester, 18, is then
`carried throuh a series of reaction steps analogous to
`those discussed above in Reaction Sequence 1 to pro-
`duce the compounds of Formula I.
`The acid, 17,
`is decarboxylated by heating to the
`corresponding quinoline, l9, which is then converted to
`the quinoline-2-carboxaldehyde, 20, by the Vilsmeier-
`Haack formylation reaction (see A. Vilsmeier, et al,
`Ber., 60:119 (1927)).
`The aldehyde, 20, is then carried through a series of
`reaction steps analogous to those discussed above in
`Reaction Sequence 1
`to produce the compounds of
`Formula I.
`
`In the ring—opened dihydroxy acid form, compounds
`of the present invention react to form salts with phar-
`maceutically acceptable metal and amine cations
`formed from organic and inorganic bases.
`The term “pharmaceutically acceptable metal cat-
`ion” contemplates positively charged metal ions de-
`rived from sodium, potassium, calcium, magnesium,
`aluminum, iron, zinc and the like.
`
`50
`
`55
`
`60
`
`65
`
`The term “pharmaceutically acceptable amine cat-
`ion” contemplates the positively charged ions derived
`from ammonia and organic nitrogenous bases strong
`enough to form such cations. Bases useful for the forma-
`tion of pharmaceutically acceptable nontoxic base addi-
`tion salts of compounds of the present invention form a
`class whose limits are readily understood by those
`skilled in the art. (See, for example, Berge, et al, “Phar-
`maceutical Salts,” J. Pharm. Sci., 66:1—19 (1977)).
`The free acid form of the compound may be regener-
`ated from the salt, if desired, by contacting the salt with
`a dilute aqueous solution of an acid such as hydrochlo-
`ric acid.
`
`The base addition salts may differ from the free acid
`form of compounds of this invention in such physical
`characteristics as melting point and solubility in polar
`solvents, but are considered equivalent to the free acid
`forms for purposes of this invention.
`The compounds of this invention can exist in unsol-
`vated as well as solvated forms. In general, the solvated
`forms, with pharmaceutically acceptable solvents such
`as water, ethanol, and the like, are equivalent to the
`unsolvated forms for purposes of this invention.
`
`Mylan Exhibit 1021, Page 6
`
`Mylan Exhibit 1021, Page 6
`
`

`
`4,761,419
`
`TABLE 1
`
`
`
`X
`—CH=CH-'
`—CH= Cl-1-
`
`R3 R4 R5 R5
`R2
`R;
`H Cl H H
`4-Fluorophenyl —CI-I3
`4-Fluorophenyl
`--CI-l(Cl-I3); H Cl H H
`
`CS1 IC5o
`p.Mole/Liter
`0.35
`0.032
`
`For preparing pharmaceutical compositions from the
`compounds described by this invention, inert, pharma-
`ceutically acceptable carriers can be either solid or
`liquid. Solid form preparations include powders, tab-
`lets, dispersible granules, capsules, cachets, and supposi-
`tories.
`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 finely divided active compound. In
`tablets, the active compound is mixed with the carrier
`having the necessary binding properties in suitable pro-
`portions and compacted in the shape and size desired.
`For preparing suppository preparations, a low-melt-
`ing wax such as a mixture of fatty-acid glycerides and
`cocoa butter is first melted, and the active ingredient is
`dispersed homogeneously therein, as by stirring. The
`molten homogeneous mixture is then poured into con-
`venient sized molds and allowed to cool and solidify.
`The powders and tablets preferably contain 5 to
`about 70% of the active ingredient. Suitable solid carri-
`ers are magnesium carbonate, magnesium stearate, talc,
`sugar,
`lactose, pectin, dextrin,
`starch,
`tragacanth,
`methyl cellulose, sodium carboxymethyl cellulose, a
`low-melting wax, cocoa butter, and the like.
`
`45
`
`50
`
`55
`
`65
`
`vided into unit doses containing appropriate quantities
`of the active component. The unit dosage form can be a
`packaged preparation, the package containing discrete
`quantities of preparation, for example, packeted tablets,
`capsules, and powders in vials or ampoules. The unit
`dosage form can also be a capsule, cachet, or tablet itself
`or it can be the appropriate number of any of these
`packaged forms.
`'
`In therapeutic use as hypolipidemic or hypocholes-
`terolemic agents, the compounds utilized in the pharma-
`ceutical method of this invention are administered to
`the patient at dosage levels of from 0 mg to 600 mg per
`day. For a normal human adult of approximately 70 kg
`or body weight, this translates to a dosage of from about
`0.5 mg/kg to about 8.0 mg/kg of body weight per day.
`The dosages, however, may be varied depending
`upon the requirements of the patient, the severity of the
`condition being treated, and the compound being em-
`ployed. Determination of optimum dosages for a partic-
`ular situation is within the skill of the art.
`The following examples illustrate particular methods
`for preparing compounds in accordance with this in-
`vention. These examples are illustrative and are not to
`be read as limiting the scope of the invention as it is
`defined by the appended claims.
`
`Mylan Exhibit 1021, Page 7
`
`11
`The compounds of this invention are useful as hypo-
`cholesterolemic or hypolipidemic agents by virtue of
`' their ability to inhibit the biosynthesis of cholesterol
`through inhibition of the enzyme 3-hydroxy-3-methyl-
`glutaryl-coenzyme A reductase (HMG-CoA reduc-
`tase).
`The ability of compounds of the present invention to
`inhibit the biosynthesis of cholesterol was measured by
`a method (designated CSI screen) which utilizes the
`procedure described by R. E. Dugan et al, Archiv. Bio-
`chem. Biophys, (1972), 152, 21-27. In this method, the
`level of HMG-CoA enzyme activity in standard labora-
`tory rats is increased by feeding the rats a chow diet
`containing Scholestyramine for four days, after which
`the rats are sacrificed.
`
`The rat livers are homogenized, and the incorpora-
`tion of l4C-acetate into nonsaponifiable lipid by the rat
`liver homogenate is measured. The micromolar concen-
`tration of compound required for 50inhibition of sterol
`synthesis over a one-hour period is measured, and ex-_
`pressed as an IC50 value.
`A
`The activities of several representative examples of
`compounds in accordance with the present invention
`appear in Table 1.
`
`5
`
`10
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`15
`
`20
`
`12
`The term “preparation” is intended to include the
`formulation of the active compound with encapsulating
`material as 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. Similarly, cachets are included. Tablets, pow-
`ders, cachets, and capsules can be used as solid dosage
`forms suitable-for oral administration.
`Liquid form preparations include solutions, suspen-
`sions, and emulsions. As an example may be mentioned
`water or water-propylene glycol solutions for paren-
`teral injection. Liquid preparations can also be formu-
`lated in solution in aqueous polyethylene glycol solu-
`tion. Aqueous solutions for oral use can be prepared by
`dissolving the active component in water and adding
`suitable colorants,
`flavoring agents, stabilizers, and
`thickening agents as desired. Aqueous suspensions for
`oral use can be made by dispersing the finely divided
`active component in water with viscous material, i.e.,
`natural or synthetic gums, resins, methyl, cellulose,
`sodium carboxymethyl cellulose, and other well-known
`suspending agents.
`Preferably, the pharmaceutical preparation is in unit
`dosage form. In such form, the preparation is subdi-
`
`Mylan Exhibit 1021, Page 7
`
`

`
`13
`
`EXAMPLE 1
`
`4,761,419
`
`Preparation of
`[4a,6B(E)]-6[-2-[6-Chloro-4-(4-fluorophenyl)-2-methyl-
`3-quinolinyl]ethenyl]-tetrahydro-4-hydroxy-2H-pyran-
`2-one
`
`Step 1—Preparation of
`(2-amino-5-chlorophenyl)-(4-fluorophenyl)methanone
`
`p-Fluorobenzoyl chloride (154.0 g, 2.4 equivalents)
`was heated to 120° C. in a 3-neck, 2-liter round-bottom
`flask. p-Chloroaniline (51.63 g) was added in portions
`with stirring and evolution of HC1 gas. After addition
`was complete, the temperature was raised to 180° C.
`and zinc chloride (66.19 g, 1.2 equivalents) was added in
`small portions. The resulting mixture was heated to 205°
`C. for three hours and then allowed to cool to about
`150° C. Hydrochloric acid (400 4mL, M) was added and
`the mixture was heated to reflux and the hot hydrochlo-
`ric acid decanted. This process was repeated twice to
`remove excess p-fluorobenzoic acid.
`Concentrated hydrochloric acid (500 mL) and glacial
`acetic acid (500 mL) were added and the resulting mix-
`ture was heated under reflux for twenty hours. The
`dark brown mixture was cooled, concentrated under
`vacuum, and partitioned between dichloromethane and
`1M sodium hydroxide solution. The organic layer was
`washed with 1M hydrochloric acid, dried over anhy-
`drous magnesium sulfate, and evaporated to give 57.3 g
`of a brown solid. This material was recrystallized from
`hexanes to yield 3.65 g of (2-amino-5-ch1orophenyl)(-
`4fluorophenyl)methanone as bright yellow needles.
`Proton NMR spectrum (CDCI3): 6 7.7-7.4, (multi-
`plet, 2 protons), 6 7.3-6.9 (multiplet, 4 protons), 6 6.55
`(doublet, 1 proton), 6 6.2 (broad singlet, 2 protons).
`
`Step 2—Preparation of
`6-chloro-4-(4-fluorophenyl)-2-methyl-3-quinolinecar-
`boxylic acid, ethyl ester
`
`To a solution of (2-amino-5-chlorophenyl)-(4-fluoro-
`phenyl)methanone (10.0 g) in 100 mL of toluene was
`added 8.3g (1.6 equivalents) of ethyl acetoacetate and
`0.30 g of p-toluenesulfonic acid. The resulting mixture
`was heated to reflux with azeotropic removal of water.
`After one and one-half hours, the mixture was cooled to
`room temperature and concentrated under vacuum.
`The crude product was purified by flash chromatogra-
`phy over silica gel to give 10.18 g of 6-chloro-4-(4-
`fluorophenyl)-2-methyl-3-quinolinecarboxylic
`acid,
`ethyl ester as an off-white solid.
`1
`Proton NMR spectrum (CDCI3): 6 7.9 doublet,
`proton), 6 7.7-7.0 (multiplet, 6 protons), 6 4.1 (quartet,
`2 protons), 6 2.7 (singlet, 3 protons), 6 1.0 (triplet, 3
`protons).
`
`Step 3—Preparation of
`. 6-chloro-4-(4-fluorophenyl)-2-methyl-3-quino1ineme-
`thanol
`
`6-Chloro-4-(4-fluorophenyl)-2-methyl-3-
`quinolinecarboxylic acid, ethyl ester (39.51 g) was dis-
`solved in 400 mL of dry dichloromethane in a 2-liter,
`3-neck, round-bottom flask under a nitrogen atmo-
`sphere. The solution was cooled to —78° C. and a solu-
`tion of diisobutyl aluminum hydride (2.5 equivalents) in
`dichloromethane was added dropwise with stirring.
`After complete addition,
`the mixture was stirred at
`—78° C. for one hour.
`Sodium sulfate (1.65 g, 2.5 equivalents) in 50 mL of
`water was then added and the cooling bath was re-
`
`14
`moved. The resulting gelatinous mass was stirred for
`one-half hour and then filtered through Celite® and
`sand. The solid was washed with hot ethyl acetate and
`the combined filtrates were dried and evaporated to
`yield 28.72 g of 6-chloro-4-(4-fluorophenyl)-2-methyl3-
`quinolinemethanol as an off-white solid, mp 173°-175°
`C.
`
`10
`
`15
`
`20
`
`25
`
`30
`
`35
`
`45
`
`50
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`55
`
`60
`
`65
`
`1
`Proton NMR spectrum (CDCI3) 6 7.8 doublet,
`proton), 6 7.(doublet of doublets,
`1 proton), 6 7.2-7.0
`(multiplet, 5 protons), 64.5 (singlet, 2 protons), 6 2.8
`(singlet, 3 protons), and 2.1 (broad singlet,
`l proton).
`
`Step 4-Preparation of
`6-chloro-4-(4-fluorophenyl)-2-methyl-3-quinolinecab
`boxaldehyde
`
`To a solution of oxalyl chloride (8.85 mL, 1.1 equiva-
`lents) in 150 mL of dry dichloromethane at —78° C.
`under nitrogen, was added dropwise a solution of di-
`methylsulfoxide (14.2 mL, 2.2 equivalents) in 125 mL of
`dichloromethane. After complete addition, the mixture
`was stirred for five minutes at —78° C. and then a solu-
`tion
`of
`6-chloro-4-(4-fluorophenyl)-2-methyl-3-
`quinolinemethanol (27.8 ) in 100 mL of dichlorometh-
`ane and 50 mL of dimethylsulfoxide was added drop-
`wise. This solution was stirred at —78° C. for one hour,
`after which triethylamine (64.1 mL, 5.0 equivalents)
`was added. The cooling bath was removed and 400 mL
`of saturated ammonium chloride solution was added to
`the reaction mixture.
`
`The organic layer was separated and the aqueous
`layer was extracted with additional dichloromethane.
`The combined organic layers were washed with water
`and then brine solution. The organic solution was dried
`over anhydrous magnesium sulfate and evaporated to
`give 27.63 g of 6-chloro-4-(-fluorophenyl)-2-methyl-
`Squinolinecarboxaldehyde as a tan solid.
`Proton NMR spectrum (CDCI3) 6 9.9 (singlet, 1 pro-
`ton), 6 7.95 (doublet, l proton), 6 7.7 (doublet of dou-
`blets, 1 proton), 6 7.5-7.1 (multiplet, 5 protons), and 6
`2.9 (singlet, 3 protons).
`
`Step 5-—Preparation of
`(E)-3-[6-chloro-4-(-fluorophenyl)-2-methyl-3-quinolyl]-
`propenoic acid, methyl ester
`
`6-Chloro-4-(4—fluorophenyl)-2-methyl-3-
`quinolinecarboxaldehyde (3.60 g) and methyl(triphenyl-
`phosphoranylidene) acetate (4.42 g, 1.1 equivalents)
`were stirred in 100 mL of dichloromethane under a
`nitrogen atmosphere for five hours at room tempera-
`ture. The mixture was concentrated under vacuum and
`the residue was purified by flash chromatography on
`silica gel to give 3.98 g of (E)-3-[6-chloro-4-(-fluoro-
`phenyl)2-methyl-3-quinolyl]propenoic
`acid, methyl
`ester as a white solid.
`
`1
`Proton NMR spectrum (CDCI3): 6 7.9 (doublet,
`proton), 7.7-7.5 (multiplet, 2 protons), 7.3-7.0 (multi-
`plet, 5 protons), 6 .8 (doublet, 1 proton), 6 3.70 (singlet,
`3 protons), and 2.8 (singlet, 3 protons).
`
`Step 6—Preparation of
`(E)--
`-3-[6-chloro-4-(4-fluoro-phenyl)-2—methy1-3-quinolyl]-
`propenol
`
`To a solution of (E-3-[6-chloro-4-(-fluorophenyl)-2-
`methyl-3-quinolyl]propenoic acid, methyl ester (7.99 g)
`in 150 mL of dry dichloromethane at —78° C. was
`added dropwise 2.5 equivalents of a solution of
`
`Mylan Exhibit 1021, Page 8
`
`Mylan Exhibit 1021, Page 8
`
`

`
`15
`diisobutylaluminum hydride (DIBAL, 1M in dichloro-
`methane) under a nitrogen atmosphere. After complete
`addition, the cooling bath was removed and the reac-
`tion was quenched by the addition of 8.0 g (2.5 equiva-
`lents) of a saturated solution of sodium sulfate. The
`resulting mixture was filtered through Celite® and
`sand. The solids were washed with hot ethyl acetate
`and the combined filtrates were dried over anhydrous
`magnesium sulfate and evaporated to yield 6.59 of (E)-
`3-[6-chloro-4-(4-fluorophenyl)-2-methyl-3-quinolyl]-
`propenol as a white solid.
`1
`Proton NMR spectrum (CDC13) 8 7.9 (doublet,
`proton), 7.5 (doublet of doublets,
`1 proton), 8 7.3-7.0
`(multiplet, 5 protons), 8 6.4 (doublet,
`1 proton), 8 5.6
`(doublet of triplets,
`l proton), 8 4.1 (broad singlet, 2
`protons), 8 2.7 (singlet, 3 protons), and 8 1.5 (broad
`triplet, 1 proton).
`
`Step 7—Preparation of
`(E)-3-[6-chloro-4-(4-fluorophenyl)-2-methyl-3-
`quinolyl]propenal
`
`A solution of oxalyl chloride (2.28 mL, 1.3 equiva-
`lents) in 100 mL of dichloromethane was cooled to
`—78° C. under a nitrogen atmosphere. A solution of
`dimethylsulfoxide (3.67 mL, 2.6 equivalents) in 75 mL
`of dichloromethane was added dropwise with stirring.
`Five minutes after addition was complete, a solution of
`(E)-3-[6-chloro-4-(4-fluorophenyl)-2-methyl-3-
`quinolyl]-propenol (6.59 g) in 75 mL of dichlorometh-
`ane was added dropwise. This solution was stirred at
`—78° C. for three-quarters of an hour and then 1mL (5.0
`equivalents) of triethylamine was added.
`The cold bath was removed and the reaction was
`quenched by the addition of 50 mL of saturated ammo-
`nium chloride solution. The organic layer was sepa-
`rated and the aqueous layer was extracted with dichlo-
`romethane. The combined organic layers were washed
`with water and then brine soluion, dried over anhy-
`drous magnesium sulfate, and evaporated to yield 6.14 g
`of
`(E-3-[6-chloro-4-(-fluorophenyl)-2-methyl-3-
`quinolyl]propenal as an orange solid.
`1
`Proton NMR spectrum (CDCl3): 8 9.4 (doublet,
`proton), 8 7.9 (doublet, 1 proton), 8 7.7-7.0 (multiplet, 7
`protons), 6.1 (doublet of doublets,
`1 proton), and 2.8
`(singlet, 3 protons).
`
`Step 8—Preparation of
`(E)7—[3-[6-chloro-4-(4-fluoro-phenyl)-2-methyl-3-
`quinolyl]-5-hydroxy-3-oxo-6-heptenoic acid, ethyl ester
`
`A solution of ethyl acetoacetate (3.1 g, 1.1 equiva-
`lents) in 30 mL of anhydrous tetrahydrofuran was
`added dropwise with stirring to a suspension of sodium
`hydride (0.64 g, 1.2 equivalents) in anhydrous tetrahy-
`drofuran at 0° C. under nitrogen. The resulting mixture
`was stirred at 0° C. for ten minutes, after which n-butyl
`lithium (10.13 mL, 2M solution in tetrahydrofuran) was
`added dropwise. The resulting orange solution was
`stirred for an additional ten minutes and then it was
`cooled to —-78° C. A solution of (E)-3-[6-chloro-4-(4-
`fluorophenyl)-2-methyl-3-quinolyl]propenal (7.20 g) in
`90 mL of anhydrous tetrahydrofuran was added drop-
`wise. The resulting solution was stirred for twenty min-
`utes before the reaction was quenched by the addition
`of 10 mL of glacial acetic acid and the cooling bath was
`removed. -
`
`The pale orange solution was then stirred at room
`temperature for two