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`E®&flJE@WH;0J
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`§11lAIILL_£. GLOLMIEJB
`
`UNITED STATES DEPARTMENT OF COMMERCE
`
`United States Patent and Trademark Office
`
`February 12, 2014
`
`THIS IS TO CERTIFY THAT ANNEXED IS A TRUE COPY FROM THE
`RECORDS OF THIS OFFICE OF THE FILE WRAPPER AND CONTENTS
`OF:
`
`APPLICATION NUMBER: 07/233, 752
`
`FILING DATE: August 19, 1988
`
`iii”
`
`
`
`By Authority of the
`
`Under Secretary of Commerce for Intellectual Property
`and Director of the United States Patent and Trademark Office
`
`
`
`M. TARVER
`
`Certifying Officer
`
`Mylan Exhibit 1003, Page,_\ I a
`
`
`
`"'
`
`'
`
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`
`Mylan Exhibit 1003, Page 1
`
`
`
`
`
`
`
`RIALNUMBER -
`07/233;752
`-
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`r
`‘ NG DATE CLASS
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`JAPAN} MIKIO SUZUKI; %umAeAsH1,_uAPANi
`xyxno suJ1KAwA, FUNABASHI:
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`JAPAN; MITSUAKI SAKASHITA; MINANISAITAMA-G:
`Jil-\P-'_AS\l$ MA-SAKI K.IT‘AH/WA: MINAMISAI‘TA'Mi\-G:
`JAPAN.
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`ttéomrluulne bATA********i***:********
`VERIFIED
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`'ER'FIED
`JAPAN
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`207224/1937A‘
`JAPAN‘
`15585i]9E_8
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`08120/87
`‘D1/26i8‘8.
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`08LONw FISHER: SPIVAK.
`
`*MQ‘CLELLfiND E MAIER
`1?3V 3. JEFF; DAMIS Hwv.
`CRYSTAL sa. FIVE-STE. 400
`.ARLINGTONr VA 22202
`
`ATTORNEN/‘S
`DOCKET No,
`
`V5».
`
`U.$. DEPT of COMM
`
`
`
`
`
`Mylan Exhibit 1003, Page 2
`
`
`
`PATENT APRLICATION SERIAL NO.
`
`233752
`
`U.S. DEPARTMENT OF COMMERCE
`PATENT AND TRADEMARK OFFICE
`FEE RECORD SHEET
`
`080
`
`OBKZEKSB
`
`233?52
`
`1 101
`
`éE0.fi0 EK
`
`D
`
`PTO—1556
`
`(5/87)
`
`Mylan Exhibit 1003, Page 3
`
`
`
`Mylan Exhibit 1003, Page 3
`
`
`
`
`
`= I-
`
`
`
`DOCS.
`
`'_
`
`49—111—o
`
`
`
`.TRANSIvIITT .
`INVENTOHISI
`
`FILING IS THE PATENT APPLICATION OF
`_, 55%
`IHIRO FUJIKAWA ET AL
`
`FOR
`
`QUINOLINE TYPE MEVALONOLACTONES
`
`ENCLOSED ARE
`
`[3 -__.________. SHEETS OF DRAWINGS.
`
`D A CERTIFIED COPY OF A___,._________________ APPLICATION.
`
`I:'] AN ASSIGNMENT OF THE INVENTION TO
`
`E:| A VERIFIED STATEMENT TO ESTABLISH SMALL ENTITY STATUS UNDER 37 CFR 1.27.
`
`LIST OF INvENTORS' NAMES AND ADDRESSES, NOTICE OF PRIORITY
`WHITE ADVANCE SERIAL NUMBER POSTCARD -
`THE FILING FEE IS CALCULATED BELOW
`
`SMALL ENTITY
`
`OTHER THAN A
`SMALL ENTITY
`
`_(COL: 2)
`(COL. 1)
`
`E _
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`|:I FILING IN NON-ENGLISH LANGUAGE
`
`[:] RECORDATION OF ASS|G_NMENT
`IF THE DIFFERENCE IN COL. 1 IS LESS THAN ZERO,
`ENTER "0" IN COL. 2
`
`150030
`I: PLEASE CHARGE MY DEPOSIT ACCOUNT NO.
`OF
`._____. A DUPLICATE COPY OF THIS SHEET IS ENCLOSED.
`
`IN THE AMOUNT
`
`THE COIYIIYIISSIONER IS HEREI3Y«AUTHORIzED TO CHARGE ANY ADDITIONAL FEES WHICH
`MAY BE REOUIRED,'OR CREDIT ANY OVERPAYMENT TO ACCOUNT NO. _.£;<m0___ .A
`DUPLICATE COPY OF THIS SHEET IS ENCLOSED.
`
`A CHECK IN THE AMOUNT OF _§§_0_-_9.9___ TO COVER THE FILING FEE IS ENCLOSED.
`
`CRYSTAL SOUARE F|VE——SUITE 400
`1755 S. JEFFERSON DAVIS HIGHWAY
`ARLINGTON, VIRGINIA 22202
`703-521-5940
`
`OBLON, FISHER, SPIVAK,
`MCCL
`AND AND MAIER, P.C.
`
`
`
`Norman F. 'OblOn
`ATTORNEY OF RECORD
`REGISTRATION NO,
`24 , 61 8
`Samuel H. Blech
`32,082
`Registration NO:
`Mylan Exhibit 1003, Page 4
`
`
`
`Mylan Exhibit 1003, Page 4
`
`
`
`
`
`
`
`Our Ref.: NC-ll5
`
`9*
`QUINOLINE TYPE MEVALONOLACTONES§
`
`The present invention relates to novel
`
`mevalonolactones having a quinoline ring, processes for
`
`their production, pharmaceutical compositions containing
`
`them and their pharmaceutical uses particularly as
`
`anti-hyperlipidemic, hypolipoproteinemic and
`
`anti—atherosclerotic agents, and intermediates useful for
`
`their production and processes for the production of such
`
`intermediates.
`
`10
`
`Some fermentation metabolic products such as
`
`compactine, CS—5l4, Mevinolin or semi—synthetic
`
`derivatives or fully synthetic derivatives thereof are
`
`known to be inhibitors against HMG—CoA reductase which is
`
`a rate limiting enzyme for cholesterol biosynthesis.
`
`(A.
`
`15
`
`Endo J. Med Chem., 28(4) 401 (1985))
`CS+5l4 and Mevinolin have been clinically proved to be
`
`potentially useful anti-hyperlipoproteinemic agents, and
`
`they are considered to be effective for curing or
`
`preventing diseases of coronary artery sclerosis or
`
`20
`
`atherosclerosis.
`
`(IXth Int. Symp. Drugs Affect. Lipid
`
`Mylan Exhibit 1003, Page 5
`
`
`
`
`
`Mylan Exhibit 1003, Page 5
`
`
`
`
`
`' Metab., 1986, p30, p31, p66)
`
`_ 2 _
`
`However, with respect to fully synthetic derivatives,
`
`particularly hetero aromatic derivatives of inhibitors
`
`against HMG—CoA reductase,
`
`limited information is
`
`disclosed in the following literatures:
`
`WPI ACC NO. 84-158675, 86—O28274, 86—0988l6,
`
`86-332070, 87-124519, 87-220987, 88-07781, 88—008460,
`
`88-091798 and 88-ll2505.
`
`The present inventors have found that mevalonolactone
`
`l0
`
`derivatives having a quinoline ring,
`
`the corresponding
`
`dihydroxy carboxylic acids and salts and esters thereof
`
`have high.inhibitory activities against cholesterol
`
`biosynthesis wherein HMG—CoA reductase acts as a rate
`
`limiting enzyme.
`
`The present invention has been
`
`15
`
`accomplished on the basis of this discovery.
`
`The novel mevalonolactone derivatives of the present
`
`invention are represented by the following formula I:
`
`3
`R \
`
`R4
`
`20
`
`R5
`
`R2
`
`Y-Z
`
`RIX
`
`N
`
`R5
`
`2
`3
`» wherein R1, R , R , R
`
`4
`
`and R
`
`6 are independently hydrogen,
`
`alkyl, C3_6 cycloalkyl, Cl_3 alkoxy, n-butoxy,
`C
`l-6
`i—butoxy, sec-butoxy, R7R8N— (wherein R7 and R8 are
`
`25
`
`independently hydrogen or Cl_3 alkyl),
`
`trifluoromethyl,
`
`trifluoromethoxy, difluoromethoxy, fluoro, chloro, bromo,
`
`Mylan Exhibit 1003, Page 6
`
`
`
`Mylan Exhibit 1003, Page 6
`
`
`
`yvphenyl, phenoxy, benzyloxy, hydroxy,
`
`._3_.
`
`QOR
`diphenyl—t—butylsilyloxy, hydroxymethyl or —O(CH
`(wherein R19 is hydrogen or Cl_3 alkyl, and 2
`is 1,
`2 or
`
`trimethylsilyloxy,
`l9
`
`3); or when located at the ortho position to each other,
`3
`4
`,
`5%%A¢a¢”9
`5% R1 and R2, or R
`and R.
`togethea;form -CH=CH—CH=CH-; or
`when located/§ the ortho position to each other, R1 and
`fl
`.;//W///
`R2 togethe5Lform —OC(Rl5)(Rl6)O— (wherein R15 and R16 are
`independently hydrogen or Cl_3 alkyl); Y is —CH 2"!
`—CH2CH2-,
`-CH=CH-,
`-CH2—CH=CH- or —CH=CH—CH2—; and Z
`-Q-CHZWCHZ-CO2Rl2,
`
`A;
`
`l0
`
`is
`
`r
`
`0 /V 0
`(
`
`‘L
`
`R11
`/
`
`H0
`
`I
`
`0
`
`/
`
`T
`
`A
`
`‘
`
`.
`
`’
`
`.15
`
`_
`
`R
`
`R“
`r
`3
`
`0
`
`0
`
`\//
`I
`
`c02R*=
`‘
`
`R‘7
`[3>T’//
`I
`0\\7///
`.‘
`or
`-C(ORl3)2— or -CH(OH)-; w is -c(o)—,
`(wherein Q is —c(o)—,
`—C(ORl3)2- or -C(Rll)(OH)-7 R11 is hydrogen or Cl_3 alkyl;
`R
`is hydrogen or R
`(wherein Rl4 is physiologically
`
`l2
`
`l4
`
`20
`
`hydrolyzable alkyl or M (wherein M is NH
`
`4/
`
`sodium,
`
`potassium,
`
`l/2 calcium or a hydrate of lower alkylamine,
`
`di—lower alkylamine or tri—lower alkylamine));
`
`two R13 are
`
`V
`
`1
`
`independently primary or secondary Cl_6 alkyl; or two R13
`R17
`l8
`together form —(CH2)2— or ~(CH2)
`and R
`are
`3‘?
`independently hydrogen or Cl_3 alkyl; and R5 is
`
`hydrogen, Cl_6 alkyl, C2_3 alkenyl, C3_6 cycloalkyl,
`9
`'
`"V
`_<§j{R (wherein R9 is hydrogen, Cl_4 alkyl, Cl_3
`
`i
`
`.
`
`25
`
`30
`
`Mylan Exhibit 1003, Page 7
`
`
`
`Mylan Exhibit 1003, Page 7
`
`
`
`_5_
`
`ethyl, n—propyl,
`
`i~propyl, n—butyl,
`
`i—butyl, sec—butyl,
`
`t—butyl, n-pentyl and n—hexyl.
`C3_6 cycloalkyl for R5 includes, for example,
`cyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl.-
`C2_3 alkenyl for R5 includes, for example, vinyl and
`
`i—propenyl.
`
`.
`Phenyl-(CH2)m- for R
`includes, for example, benzyl,
`B-phenylethyl and y-phenylpropyl.
`5
`
`5
`
`Phenyle(CH2)nCH(CH3)— for R
`
`includes, for example,
`
`5
`
`10
`
`a-phenylethyl and a—benzylethyl.
`8
`
`C
`
`1-3
`
`alkyl for R7 and R
`
`includes, for example,
`
`methyl, ethyl, n¥propyl and i-propyl.
`
`Further,
`
`these compoundsnmy have at least one or two
`
`asymmetric carbon atoms and may have at least two to four
`
`15 optical isomers.
`
`The compounds of the formula I_include
`
`all of these optical isomers and all of the mixtures
`
`thereof.
`
`Among compounds having carboxylic acid moieties
`
`falling outside the definition of -COZR
`
`12 of the
`
`20
`
`carboxylic acid moiety of substituent Z of the compounds
`
`of the present invention,
`
`those which undergo
`
`physiological hydrolysis, after intake,
`
`to produce the
`
`corresponding carboxylic acids (compounds wherein the
`-CO2Rl2 moiety is -CO2H) are equivalent to the compounds
`
`25
`
`of the present invention.
`Now, preferred substituents of the compounds of the
`
`present invention will be described.
`
`Mylan Exhibit 1003, Page 8 y
`
`
`
`
`
`Mylan Exhibit 1003, Page 8
`
`
`
`
`
`-6-
`
`In the following preferred, more preferred still
`
`further perferred and most preferred examples,
`
`the
`
`numerals for the positions of the substituents indicate
`
`the positions on the quinoline ring.
`
`For example, N‘
`
`shown by e.g. l' or 2'
`
`indicates the position of the
`
`substituent on the phenyl substituted at the 4—position of‘
`
`the quinoline ring (the carbon connected to the quinoline
`
`ring is designated as 1').
`
`The meanings of the respective
`
`substituents are the same as the above—mentioned meanings.
`l
`2
`Preferred substituents for R , R
`
`and R6 are hydrogen,
`
`l0
`
`fluoro, chloro, bromo, Cl_3 alkyl, Cl_3 alkoxy, C3_6
`cycloalkyl, dimethylamino, hydroxy, hydroxymethyl,
`
`hydroxyethyl,
`
`trifluoromethyl,
`
`trifluoromethoxy,
`
`difluoromethoxy, phenoxy and benzyloxy.
`
`15
`
`Further, when R6 is hydrogen, it is preferred that R1
`and R2 together form methylenedioxy.
`
`As preferred examples for R3 and R4, when R4 is
`hydrogen, R3 is hydrogen, 3'—fluoro, 3'-chloro, 3'—methyl,
`
`4'-methyl, 4'-chloro and 4'—fluoro.
`
`20
`
`Other preferred combinations of R
`
`3 and R4
`
`include
`
`3'—methyl-4'-chloro, 3',5'-dichloro, 3',5'—difluoro,
`
`3',5'-dimethyl and 3'—methyl-4'-fluoro.
`
`Preferred examples for R5 include primary and
`
`secondary Cl_6 alkyl and C3_6 cycloalkyl.
`
`25
`
`Preferred examples for Y include -CH2—CH2— and
`-CH=CH—.
`
`Preferred examples for Z include
`
`Mylan Exhibit 1003, Page 9
`
`Mylan Exhibit 1003, Page 9
`
`
`
`_.8_
`
`V 6,7-difluoro, 6,8—difluoro, 6,7—methylenedioxy,
`
`6,8—dichloro, 5,8-dimethyl, 6,8—dimethyl, 6,7—dimethoxy,
`
`6,7—diethoxy, 6,7-dibromo or 6,8—dibromo.
`
`l
`when R , R
`
`2
`
`6
`
`and R
`
`are not hydrogen,
`
`they together
`
`represent 5,7-dimethoxy—8-hydroxy, 5,8-dichloro-6-hydroxy,
`
`6,7,8—trimethoxy, 6,7,8-trimethyl, 6,7,8-trichloro,
`
`5—fluoro—6,8-dibromo or 5—chloro—6,8—dibromo.
`
`As more preferred examples for R
`
`3
`
`4
`and R , when R
`
`3
`
`is
`
`hydrogen, R4
`
`is hydrogen, 4'—methyl, 4'-chloro or
`
`l0
`
`4'-fluoro.
`
`when both R
`
`3
`
`and R
`
`4
`
`are not hydrogen,
`
`they
`
`together represent 3',5'-dimethyl or 3'—methyl—4'—fluoro.
`
`As more preferred examples for R5
`5
`preferred examples of R may be mentioned.
`
`,
`
`the above-mentioned
`
`As preferred examples for Y,
`
`-CH2-CH2— and (E)——CH=CH—
`
`15
`
`may be mentioned. As more preferred examples for Z,
`
`the
`
`above preferred examples for Z may be mentioned.
`
`Now, still further preferred substituents of the
`
`compounds of the present invention will be described. As
`1
`2
`6
`2
`6
`examples for R , R
`and R , when both R
`
`and R
`
`are
`
`_20
`
`hydrogen, R1 is hydrogen, 6—methyl, 6—ethyl,
`
`6—trifluoromethyl, 6—hydroxy, 6—methoxy, 6—chloro,
`
`6—bromo, 6-n-butyl and 7—dimethylamino.
`2
`when only R6 is hydrogen, R1 and-R represent
`
`6,8—dichloro, 5,8—dimethyl, 6,8—dimethyl, 6,7—dimethoxy,
`
`25
`
`6,7—diethoxy, 6,7-dibromo, 6,8—dibromo, 6,7-difluoro and
`
`6,8—difluoro.-A
`
`_As still further preferred examples for R3 and R4,
`
`Mylan Exhibit 1003, Page 10 I
`
`
`
`Mylan Exhibit 1003, Page 10
`
`
`
`
`
`-9-
`
`when R3 is hydrogen, R4 is hydrogen, 4'—chloro or
`3
`
`4'—fluoro, or R
`
`and R4 together represent
`
`3'~methyl-4f-fluoro.
`Still further preferred examples for R
`
`5
`
`‘include ethyl,
`
`n-propyl,
`
`i-propyl and cyclopropyl.
`
`Still further preferred examples for Y include
`
`(E)——CH=cH—.
`
`As still further preferred examples for Z,
`
`the
`
`above-mentioned preferred example for Z may be mentioned.
`Now,
`the most preferred substituents for the compounds
`
`l0
`
`of the present invention will be described.
`
`and R6, when
`As the most preferred examples for R1, R
`both R2 and R6 are hydrogen, R1 is hydrogen, 6—methyl or
`
`2
`
`6—chloro.
`
`15’
`
`I when only R6 is hydrogen, R
`
`1
`
`and R2 together
`
`represent, for example, 6,7-dimethoxy.
`
`As the most preferred examples for R
`
`3
`
`4
`and R , R
`
`3
`
`is
`
`hydrogen and R4 is hydrogen, 4'-chloro or 4'-fluoro.
`5
`
`The most preferred examples for R
`
`include i—propyl
`
`_2o
`
`and cyclopropyl.
`
`The most preferred example for Y may be
`
`(E)--CH=CH-.
`
`As the most preferred examples for Z,
`
`the
`
`above—mentioned preferred examples for Z may be mentioned.
`
`Now, particularly preferred specific compounds of the
`
`25
`
`present invention will be presented.
`
`The following
`
`compounds
`
`(a)
`
`to (z) are shown in the form of carboxylic
`
`acids. However,
`
`the present invention include not only
`
`Mylan AExhibit 1003, Page 117
`
`
`
`
`
`
`
`Mylan Exhibit 1003, Page 11
`
`
`
`
`
`
`
`-10..
`
`the compounds in the form of carboxylic acids but also the
`
`corresponding lactones formed by the condensation of the
`
`carboxylic acids with hydroxy at the 5-position, and
`
`sodium salts and lower alkyl esters (such as methyl,
`
`i—propyl and n—propyl esters) of the carboxylic
`ethyl,
`acids, which can be physiologically hydrolysed to the
`
`carboxylic acids.
`
`(a)
`
`(E)—3,5-dihydroxy-7—[4'—(4"—fluorophenyl)-2'-
`
`(l"-methylethyl)-quinolin-3'—yl]—hept-6-enoic acid
`
`10
`
`(b)
`
`(E)—3,5—dihydroxy-7-[4'—(4"-fluorophenyl)-2'-
`
`(l"—methylethyl)—6'-chloro—quinolin-3'—yl]—hept-6-enoic
`acid
`I
`I
`(E)-3,5—dihydroxy—7-[4'—(4"-fluorophenyl)—2'—
`
`(c)
`
`(l"-methylethyl)—6'—methyl-quinolin—3'-yl]—hept-6—enoic
`
`15
`
`acid
`
`I
`
`(d)
`
`(E)-3,5—dihydroxy—7—[4'—(4"-fluorophenyl)—2'-
`
`(l"—methylethyl)—6',7’-dimethoxy—quinolin—3'-yl]—hept—6—
`
`enoic acid
`
`(e)
`
`(E)-3,5-dihydroxy—7—[4'-(4"—fluorophenyl)-2'-
`
`V20
`
`cyclopropyl-quinolin—3'—yl]—hept—6-enoic acid
`
`(f)
`
`(E)-3,5—dihydroxy-7—[4'-(4"-fluorophenyl)—2'—
`
`cyclopropyl—6'—chloro—quinolin—3'—yl]—hept—6—enoic acid
`(g)
`(E)—3,5-dihydroxy—7—[4'-(4"-fluorophenyl)-2'-
`
`Cyclopropyl—6'—methyl-quinolin-3'-yl]—hept-6—enoic acid
`
`25
`
`(h)
`
`(E)—3,5—dihydroxyj7—[4'—(4"-fluorophenyl)—2'-
`
`cyclopropyl—6',7'-dimethoxy—quinolin-3'—yl]—hept-6—enoic
`
`acid
`
`Mylan Exhibit 1003, Page 12
`
`Mylan Exhibit 1003, Page 1
`
`
`
`Mylan Exhibit 1003, Page 12
`
`
`
`_ ll _
`
`(i)
`(E)—3,5-dihydroxy-7—[4'—(4"-chlorophenyl)-2'-
`(l"-methylethyl)—quinolin—3'—yl]—hept-6—enoic acid
`
`I
`
`(3')
`
`(E)—3,5-dihydroxy-7-[4'—(4"—chlorophenyl)—2'-
`
`(l"—methylathyl)—6'—chloro¥quinolin—3'—yl]—hept-6—enoicA
`
`acid
`
`(k)
`
`(E)-3,5-dihydroxy—7—[4'—(4"-chlorophenyl)-2'-
`
`(l"-methylethyl)-6'—methyl—quinolin—3'-yl]—hept-6eenoic
`
`acid
`
`(1)
`
`(E)-3,5—dihydroxy-7—[4'-(4"-chlorophenyl)—2'-
`
`l0
`
`(l"-methylethyl)—6',7’-dimethoxy—quinolin-3'—yl]-hept—6-
`
`enoic acid
`
`(m)
`
`(E)-3,5-dihydroxy—7—[4'—(4"—chlorophenyl)-2'-
`
`cyclopropyl—quinolin-3'-yl]-hept-6—enoic acid
`
`(n)
`
`(E)-3,5-dihydroxy—7—[4'-(4"—chlorophenyl)-2'—
`
`15
`
`cyclopropyl-6'-chloro—quinolin—3'-yl]—hept—6—enoic acid
`
`(o)
`
`(E)—3,5-dihydroxy—7—[4'—(4"—chlorophenyl)—2'-
`
`cyclopropyl-6'—methyl—quinolin-3'-yl]-hept-6—enoic acid
`
`(p)
`
`(E)-3,5—dihydroxy-7-[4'—(4"—chlorophenyl)-2'-
`
`cyclopropyl-6'7'—dimethoxy—quinolin-3'—yl]-hept-6—enoic
`acid
`7
`
`20
`
`(q)
`
`(E)-3,5—dihydroxy—7—[4'—phenyl—2'-(l"-
`
`methylethyl)-quinolin—3'—yl]—hept-6—enoic acid
`
`(r)
`
`(E)-3,5—dihydroxy-7—[4'-phenyl—2'—(l"-
`
`methylethyl)—6'—chloro-quinolin—3'—yl]—hept—6—enoic acid
`
`25
`
`(s)
`
`(E)-3,5~dihydroxy—7-[4'—phenyl—2'-(l"-
`
`methylethyl)—6'—methyl-quinolin—3'—yl]—hept—6—enoic acid
`
`(t)
`
`(E)-3,5—dihydroxy-7-[4'-pher1yl—2'—(lA' '-
`
`Mylan Exhibit 1003, Page 13
`
`
`
`Mylan Exhibit 1003, Page 13
`
`
`
`.12..
`
`methylethyl)-6',7'—dimethoxy~quinolin-3'—yl]—hept—6—enoic
`
`acid
`
`(u)
`
`(E)-3,5-dihydroxy-7—[4'—phenyl—2'—cyclopropyl—
`
`quinolin-3'-yl]-hept-6-enoic acid
`(V)
`(£3)-3,5—dihydroxy-7-[4'—phenyl—2'—cyclopropyl-6‘:-
`chloro—quinolin-3'-yl]-hept—6—enoic acid
`
`(w)
`
`(E)—3,5—dihydroxy—7-[4'-phenyl-2'-cyclopropyl-6'—
`
`methyl-quinolin-3'—yl]-hept—6—enoic acid
`
`‘(X)
`
`(E)-3,5-dihydroxy-7-[4'-phenyl—2‘-cyclopropyl—
`
`l0
`
`6',7’-dimethoxy—quinolin-3'—yl]—hept—6—enoic acid
`
`(y)
`
`(E)—3,5-dihydroxy—7—[4'-(4"—fluorophenyl)-2‘-
`
`‘(l"-methylethyl)—6'—methoxy-quinolin—3'—yl]—hept-6-enoic
`
`acid
`
`(2)
`
`(E)-3,5-dihydroxy—7—[4'-(4"-fluorophenyl)—2'—
`
`15
`
`cyclopropyl-6'—methoxy-quinolin—3'-yl]—hept—6-enoic acid
`
`The mevalonolactones of the formula I can be prepared
`
`by the following reaction scheme.
`
`The enal III can also
`
`be prepared by processes K, L and M.
`
`Mylan Exhibit 1003, Page 14
`
`
`
`Mylan Exhibit 1003, Page 14
`
`
`
`-13
`
`R4
`
`cuzon'
`
`R5
`
`;
`'
`
`B
`
`O
`N
`
`VI
`
`R3
`
`
`
`Mylan Exhibit 1003, Page 15
`
`
`
`Mylan Exhibit 1003, Page 15
`
`
`
`-14..
`
`C0z'R'2
`
`Mylan Exhibit 1003, Page 16
`
`
`
`Mylan Exhibit 1003, Page 16
`
`
`
`-15 _
`
`
`
`OH
`
`Mylan Exhibit 1003, Page 17
`
`
`
`Mylan Exhibit 1003, Page 17
`
`
`
`-16-
`
`R“
`
`R‘
`
`R=
`
`R‘
`
`R.
`
`R=
`R‘
`
`CR)
`
`v
`
`R.
`
`cRo —T———>R=
`R5
`R‘
`
`co,R==
`
`-—-—L—e
`
`l
`R‘
`
`CR)
`
`Vfll
`
`
`
`Mylan Exhibit 1003, Page 18
`
`
`
`Mylan Exhibit 1003, Page 18
`
`
`
`R‘ f\/com”
`I
`
`o
`
`R‘
`
`0N
`
`I-6
`
`Mylan Exhibit 1003, Page 19
`
`_]_7_
`
` R“
`
`RI
`
`R.
`
`n=
`
`
`
`Mylan Exhibit 1003, Page 19
`
`
`
`
`
`
`
`_20_.
`
`ethanol at a temperature of from 10 to 25°C.
`
`The free
`
`acid hereby obtained may be converted to a salt with a
`
`suitable base.
`
`‘Step H is a step for forming a mevalonolactone by the
`
`dehydration reaction of the free hydroxy acid I-2.
`dehydration reaction can be conducted in benzene or
`
`The
`
`toluene under reflux while removing the resulting water or
`
`by adding a suitable dehydrating agent such as molecular
`
`sieve.
`
`l0
`
`Further,
`
`the dehydration reaction may be conducted in
`
`dry methylene chloride by using a lactone—forming agent
`
`such as carbodiimide, preferably a water soluble
`
`carbodiimide such as
`
`15
`
`N—cyclohexyl-N’-[2'—(methylmorpholinium)ethyllcarbodiimide
`p-toluene sulfonate at a temperature of from 10 to 35°C,
`preferably from 20 to 25°C.
`
`Step J represents a reaction for hydrogenating the
`
`double bond connecting the mevalonolactone moiety and the
`
`quinoline ring. This hydrogenation reaction can be
`
`20
`
`conducted by using a catalytic amount of palladium—carbon
`
`or rhodium—carbon in a solvent such as methanol, ethanol,
`
`tetrahydrofuran or acetonitrile at a temperature of from O
`
`to 50°C, preferably from 10 to 25°C.
`
`Step K represents a reaction for the synthesis of an
`
`25
`
`a,B—unsaturated carboxylic acid ester, whereby a
`
`trans—form a,B-unsaturated carboxylic acid ester can be
`
`obtained by a so—called Horner—Wittig reaction by using an
`
`Mylan Exhibit 1003, Page 20
`
`
`
`Mylan Exhibit 1003, Page 20
`
`
`
`-21-
`
`alkoxycarbonylmethyl phosphonate.
`
`The reaction is
`
`conducted by using sodium hydride or potassium t-butoxide
`
`as the base in dry tetrahydrofuran at a temperature of
`
`from -30 to 0°C, preferably from -20 to -l5°C.
`
`Step L represents a reduction reaction of the
`
`d,B-unsaturated carboxylic acid ester to an allyl alcohol.
`
`This reduction reaction can be conducted by using various
`
`metal hydrides, preferably diisobutylaluminiumhydride,
`
`in
`
`a solvent such as dry tetrahydrofuran or toluene at a
`
`l0
`
`temperature of from -l0 to l0OC, preferably from -10 to
`
`0°C.
`
`Step M represents an oxidation reaction of the allyl
`
`alcohol
`
`to an enal. This oxidation reaction can be
`
`conducted by using various oxidizing agents, particularly
`
`l5
`
`active manganese dioxide,
`
`in a solvent such as
`
`tetrahydrofuran, acetone, ethyl ether or ethyl acetate at
`
`a temperatrue of from O to l0OOC, preferably from l5 to
`
`50°C.
`
`Step N represents a reaction for the synthesis of an
`
`20
`
`a,B—unsaturated ketone by the selective oxidation of the
`
`dihydroxy carboxylic acid ester. This reaction can be
`
`conducted by using activated manganese dioxide in a
`
`solvent such as ethyl ether,
`
`tetrahydrofuran,
`
`benzene or
`
`toluene at a temperature of from 20 to 800C, preferably
`
`25
`
`from 40 to 80°C.
`
`In addition to the compounds disclosed in Examples
`
`given hereinafter, compounds of the formulas I-2 and I-5
`
`Mylan Exhibit 1003, Page 21
`
`
`
`Mylan Exhibit 1003, Page 21
`
`
`
`_.22_
`
`given in Table l can be prepared by the process of the
`
`present
`
`invention.
`
`In Table l,
`
`i— means iso,
`
`sec— means
`
`secondary and c— means cyclo. Likewise, Me means methyl,
`
`Et means ethyl, Pr means propyl, Bu means butyl, Pent
`
`means pentyl, Hex means hexyl and Ph means phenyl.
`
`Mylan Exhibit 1003, Page 22
`
`
`
`Mylan Exhibit 1003, Page 22
`
`
`
`Table 1
`
`-23..
`
`fbj
`
`R“
`i
`R‘ i<@/1% R5
`
`0 H
`
`OH
`
`co2R1H=
`\
`%:§9::::2:)
`
`
`
`R“
`
`R“
`
`R“
`
`R‘
`
`R2
`
`R“
`
`6-0?-1e
`
`H“
`
`6—OHe
`
`6-Br
`
`V
`
`H
`
`H
`
`H
`
`4-F
`
`4-17
`
`6-F-1e
`
`8-Me
`
`4-F
`
`7-0Me
`
`8-0P1e 4-F
`
`6-Br
`
`H
`
`2-F
`
`H
`
`H
`
`H
`
`H
`
`H
`
`H
`
`6.7‘
`</_\>
`
`H
`
`H
`
`i-Pr
`
`i-Pr
`
`i-Pr
`
`i-Pr
`
`i-Pr
`
`i-Pr
`
`i-Pr
`—<i‘>
`
`H
`
`H
`
`H
`
`H
`
`H
`
`H
`
`H
`H
`
`H
`
`H
`
`64012
`
`6-C2
`
`5—0CH2Ph
`
`H
`
`H
`
`63-02
`
`6—i‘1ezN
`
`H
`
`H
`
`H
`
`H
`
`H
`
`H
`
`H
`
`H
`
`H
`
`4-F
`4-13
`
`4-Ph
`
`H
`H
`
`H
`
`i-Pr
`
`4—PhCHz H
`
`'1-Pr
`
`H
`
`H
`
`H
`
`H
`
`H
`
`H
`
`c-Pr
`
`sec—Bu
`
`V
`
`i-Pr
`
`i—Bu
`
`4-17
`
`4—F
`
`4* F
`
`4-17
`
`4-F
`
`4-F
`
`4‘F
`
`H
`
`H
`
`H
`
`H
`
`H
`
`H
`
`H
`
`C-Pent: H
`
`c-Pent H
`
`i-Pr‘
`
`H
`
`Mylan Exhibit 1003, Page 23
`
`
`
`
`
`Mylan Exhibit 1003, Page 23
`
`
`
`-24-
`
`R‘
`
`R2
`
`R3
`
`R‘
`
`R5
`
`R“
`
`6-He
`
`6—i-Pr
`
`7-Me
`
`6-0Me
`
`6-Br
`
`6—i-Pr
`
`H
`
`H
`
`H
`
`H
`
`H
`
`H
`
`4- F
`
`4- F
`
`4- F
`
`4- F
`
`4- F
`
`4- F
`
`6-02
`
`8-01’.
`
`4- F
`
`5-F
`
`6-Br
`
`4 - F
`
`H
`
`H
`
`H
`
`H
`
`H
`
`H
`
`H
`
`H
`
`H
`
`c-Pr
`
`i-Pr
`
`c-Pr
`
`c-Pr
`
`C-Pr
`
`c-Pr
`
`c-Pr
`
`H
`
`H
`
`H
`
`H
`
`H
`
`H
`
`H
`
`i-Pr
`
`8—Br
`
`i—Pr
`
`8-On‘-1e
`
`6-0:‘-1e
`
`7—0P1e
`
`4 - F
`
`5-Me
`
`7—Me
`
`4- F
`
`6-C17.
`
`7-0.0.
`
`4-"F
`
`H
`
`H
`
`H
`
`H
`
`4- F
`
`4- F
`
`6—0Me
`
`7-0:”-le
`
`H
`
`Home
`
`7-0Me
`
`4'-cg
`
`6-0:‘-1e
`
`7—0He
`
`H
`
`6-0!‘-1e
`
`7-0Me
`
`4-C2
`
`5-0318
`
`7-0:‘1e
`
`4—F
`
`H
`
`H
`
`H
`
`H
`
`H
`
`H
`
`H
`
`H
`
`H
`
`i-Pr
`
`8-Me
`
`i—Pr
`
`8—C£
`
`c—Bu
`
`c-Hex
`
`i—Pr
`
`i-Pr
`
`c-Pr
`
`c-Pr
`
`C-Pr
`
`H
`
`H
`
`H
`
`H
`
`H
`
`H
`
`H
`
`Mylan Exhibit 1003, Page 24
`
`Mylan Exhibit 1003, Page 24
`
`
`
`-25-
`
`R‘
`
`R2
`
`. R3
`
`R“
`
`R5
`
`R"'
`
`6-Me
`
`5-?-’le
`
`6-Me
`
`6-Me
`
`6—Me
`
`6-CE
`
`6-CE
`
`6—CZ
`
`63-61!
`
`6-C12
`
`H
`
`H
`
`H
`
`H
`
`H
`
`H
`
`H
`
`H
`
`H
`
`H
`
`H
`
`H
`
`H
`
`H
`
`H
`
`H
`
`H
`
`H
`
`H
`
`H
`
`H
`
`4-C2
`
`H
`
`4—C2Z
`
`4—F
`
`H
`
`4—C1Z
`
`H
`
`4—CIZ
`
`4-?‘
`
`H
`
`4—C£
`
`H
`
`4-CE
`
`4-F
`
`H
`
`H
`
`H
`
`H
`
`H
`
`H
`
`H
`
`H
`
`H
`
`H
`
`H
`
`H
`
`H
`
`H
`
`i—Pr
`
`H
`
`1-Pr
`
`c-Pr
`
`c-Pr
`
`c-Pr
`
`1-Pr
`
`1-Pr
`
`c-Pr
`
`c-Pr
`
`c-Pr
`
`1-Pr
`
`1—Pr
`
`c-Pr
`
`c-Pr
`
`c Pr
`
`H
`
`H
`
`H
`
`H
`
`H
`
`H
`
`H
`
`H
`
`H
`
`H
`
`H
`
`H
`
`H
`
`H
`
`H
`
`Mylan Exhibit 1003, Page 25
`
`
`
`Mylan Exhibit 1003, Page 25
`
`
`
`
`
`_26_
`
`Further, pharmaceutically acceptable salts such as
`
`potassium salts or esters such as ethyl esters or methyl
`
`esters of these compounds can be prepared in the same
`
`manner.
`
`The compounds of the present invention exhibit high
`
`inhibitory activities against the cholesterol biosynthesis
`
`wherein HMG-CoA reductase acts as a rate limiting enzyme,
`
`as shown by the test results given hereinafter, and thus
`
`are capable of suppressing or reducing the amount of
`
`10
`
`the compounds
`cholesterol in blood as lipoprotein. «Thus,
`of the present invention are useful as curing agents
`
`against hyperlipidemia, hyperlipoproteinemia and
`
`atheroscleosis.
`
`They may be formulated into various suitable
`
`15
`
`formulations depending upon the manner of the
`
`administration.
`
`The compounds of the present invention
`
`may be administered in the form of free acids or in the
`
`20
`
`25
`
`form of physiologically hydrolyzable and acceptable esters
`
`or lactones, or pharmaceutically acceptable salts.
`
`The pharmaceutical composition of the present
`invention is preferably administered orally in the form of
`the compound of the present
`invention per se or in the
`
`form of powders, granules,
`
`tablets or capsules formulated
`
`by mixing the compound of the present invention with a
`
`suitable pharmaceutically acceptable carrier including a
`
`binder such as hydroxypropyl cellulose, syrup, gum arabic,
`
`gelatin, sorbitol,
`
`tragacanth gum, polyvinyl pyrrolidone
`
`Mylan Exhibit 1003, Page 26 T
`
`
`
`Mylan Exhibit 1003, Page 26
`
`
`
`
`
`..
`
`or CMC-Ca, an excipient such as lactose, sugar, corn
`
`starch, calcium phosphate, sorbitol, glycine or crystal
`
`cellulose powder, a lubricant such as magnesium stearate,
`
`talk, polyethylene glycol or silica, and a disintegrator
`
`such as potato starch.
`
`However,
`
`the pharmaceutical composition of the present
`
`invention is not limited to such oral administration and
`
`it is applicable for parenteral administration.
`
`For
`
`example, it may be administered in the form of e.g. a
`
`l0
`
`15
`
`20
`
`suppository formulated by using oily base material such as
`
`lanolin or fatty acid
`cacao butter, polyethylene glycol,
`triglyceride, a transdermal therapeutic base formulated by
`
`using liquid paraffin, white Vaseline,
`
`a higher alcohol,
`
`Macrogol ointment, hydrophilic ointment or hydro—gel base
`
`material, an injection formulation formulated by using one
`
`or more materials selected from the group consisting of
`
`polyethylene glycol, hydro—gel base material, distilled
`
`water, distilled water for injection and excipient such as
`
`lactose or corn starch, or a formulation for
`
`administration through mucous membranes such as an ocular
`
`mucous membrane, a nasal mucous membrane and an oral
`
`mucous membrane.
`
`Further,
`
`the compounds of the present
`
`invention may be
`
`combined with basic ion—exchange resins which are capable
`of binding bile acids and yet not being absorbed in
`
`25
`
`gastraintestinal tract.
`
`The daily dose of the compound of the formula I is
`
`Mylan Exhibit 1003, Page 27
`
`
`
`Mylan Exhibit 1003, Page 27
`
`
`
`-28-
`
`from 0.05 to 500 mg, preferably from 0.5 to 50 mg for an
`
`adult.
`
`It is administered from once to three times per
`
`day.
`
`The dose may of course be varied depending upon the
`
`age,
`
`the weight or the condition of illness of the
`
`5
`
`patient.
`
`The compounds of the formulas II to VII are novel, and
`
`they are important
`
`intermediates for the preparation of
`
`the compounds of the formula I. Accordingly,
`
`the present
`
`invention relates also to the compounds of the formulas II
`
`10
`
`to VII and the processes for their production.
`
`Now,
`
`the present invention will be described in
`
`further detail with reference to Test Examples for the
`
`pharmacological activities of the compounds of the present
`
`invention,
`
`their Preparation Examples and Formulation
`
`15
`
`Examples. However,
`
`it should be understood that the
`
`present invention is by no means restricted by such
`
`specific Examples.
`
`PHARMACOLOGICAL TEST EXAMPLES
`
`Test A:
`
`Inhibition of cholesterol biosynthesis from
`
`20
`
`acetate in vitro
`
`Enzyme solution was prepared from liver of male wistar
`
`rat billialy cannulated and discharged bile for over 24
`
`hours. Liver was cut out at mid—dark and microsome and
`
`supernatant fraction which was precipitable with 40-80% of
`
`25
`
`saturation of ammonium sulfate (sup fraction) were
`
`prepared from liver homogenate according to the modified
`
`method of Knauss et. al.; Kuroda, M., et.
`
`al., Biochim.
`
`Mylan Exhibit 1003, Page 28
`
`
`
`Mylan Exhibit 1003, Page 28
`
`
`
`-29..
`
`Biophys. Acta, 489,
`
`ll9 (l977).
`
`For assay of cholesterol
`
`biosynthesis, microsome (0.1 mg protein) and sup fraction
`
`(1.0 mg protein) were incubated for 2 hours at 37°C in 200
`
`pl of the reaction mixture containing ATP;
`
`1 mM,
`
`5 Glutathione;
`
`6 mM, Glucose-l—phosphate; 10 mM, NAD; 0.25
`
`mM, NADP; 0.25 mM, CoA; 0.04 mM and 0.2 mM [2—l4C]sodium
`
`acetate (0.2 uci) with 4 ul of test compound solution
`
`dissolved in water or dimethyl sulfoxide.
`
`To stop
`
`reaction and saponify,
`
`1 ml of 15% EtOH—KOH was added to
`
`10-
`
`the reactions and heated at 75°C for 1 hour.
`
`Nonsaponifiable lipids were extracted with petroleum ether
`
`and incorporate
`
`d 14
`
`C radioactivity was counted.
`
`Inhibitory activity of compounds was indicated with IC50.
`
`Test B:
`
`Inhibition of cholesterol biosynthesis in
`
`15
`
`culture cells
`
`Hep G2 cells at over 5th passage were seeded to 12
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`well plates and incubated with Dulbecco's modified Eagle
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`t(DME) medium containing l0% of fetal bovine serum (FBS) at
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`37°C;
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`5% CO2 until cells were confluent for about 7 days.
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`20 Cells were exposed to the DME medium containing 5% of
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`lipoprotein deficient serum (LpDS) prepared by
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`ultracentrifugation method for over 24 hours: Medium was
`
`changed to 0.5 ml of fresh 5% LpDS containing DME before
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`assay and 10 ul of test compound solution dissolved in
`
`25 water or DMSO were added.
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`0.2 uci of
`
`l4
`[2— Clsodium
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`acetate (20 pl) was added at O hr(B—l) or 4 hrs(B—2) after
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`addition of compounds. After 4 hrs further incubation
`14
`with [2— Clsodium acetate, medium was removed and cells
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`were washed with phosphate buffered saline(PBS) chilled at~
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`4°C. Cells were scraped with rubber policeman and
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`collected to tubes with PBS and digested with 0.2 ml of
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`0.5 N KOH at 370C. Aliquot of digestion was used for
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`protein analysis and remaining was saponified with 1 ml of
`15% EtOH—KOH at 75°C for 1 hour. Nonsaponifiable lipids
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`were extracted with petroleum ether and
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`4C radioactivity
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`was counted. Counts were revised by cell protein and
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`indicated with DPM/mg protein.
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`Inhibitory activity of
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`l0
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`compounds was indicated with IC50.
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`Test C:
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`Inhibition of cholesterol biosynthesis in vivo
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`Male Sprague—Dawléy rats weighing about 150 g were fed
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`normal Purina chow diet and water ad libitum, and exposed
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`to 12 hours light/12 hours dark lighting pattern (2:00 PM
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`15
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`— 2:00 AM dark) prior to use for in vivo inhibition test
`of cholesterol biosynthesis. Animals were separated
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`groups consisting of five rats as to be average mean body
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`weight
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`in each groups. Test compounds at dosage of
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`0.02-0.2 mg/kg body weight
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`(0.4 ml/100 g body weight),
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`20
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`were dissolved in water or suspended or in 0.5% methyl
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`cellulose and orally administered at 2-3 hours before
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`midsdark (8:00 PM), while cholesterol biosynthesis reaches
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`to maximum in rats.
`As control, rats were orally
`administered only water or vehicle. At 90 minutes after
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`25
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`sample administration, rats were injected
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`intraperitoneally with 10 uCi of
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`[2—l4C]sodium acetate at
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`volume of 0.2 ml per one.
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`2 Hours later,
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`blood samples
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`were obtained and serum were separated immediately. Total
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`lipids were extracted according to the method of Folch et
`al. and saponified with EtOH-KOH. Nonsaponifiable lipids
`
`were extracted with petroleum ether and radio activity
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`5
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`incorporated into nonsaponifiable lipids was counted.
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`Inhibitory activity was indicated as percent decrease
`
`of counts in testing groups (DPM/2 ml serum/2 hours)
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`from
`
`that in control group.
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`With respect to the compounds of the present
`
`l0
`
`invention,
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`the inhibitory activities against the
`
`cholesterol biosynthesis in which HMG—CoA reductase serves
`
`as a rate limiting enzyme, were measured by the above Test
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`A and B.
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`The results are shown in Tables, 2, 2-2,
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`3 and
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`3-2. Further,
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`the results of the measurements by Test C
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`15
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`are also presented.V
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`Table 2: Inhibitory activities by Test A
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`
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`‘fi<駧3(molar concentration)
`
`Compound
`
`l0
`
`15
`
`20
`
`25
`
`30
`
`35
`
`40
`
`45
`
`50
`
`(Compounds
`of the present
`invention)
`
`1-13
`I-51
`
`I-52
`
`I-53
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`(Reference
`compounds)
`
`Mevinolin
`
`_
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`1.25 x 1o'7
`1.0
`x 1o'8
`
`7.1 x 1o"8
`
`1.9
`
`x 1o“7
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`1.4
`
`x 10-8
`
`9.6K x 1o‘9
`cs—514
`
`
`In Table 2-2,
`
`the relative activities are shown based
`
`on the activities of CS-514 being evaluated to be 1.
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`Table 2-2: Relative activities by Test A
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`
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`Relative activities
`Compound
`
`
`(Comounds of
`the present
`invention)
`
`I-l6
`
`I-116
`
`I-117
`
`I-l2O
`
`A
`
`’
`
`1.75
`
`2.25
`
`0.37
`
`3.21
`
`0.76
`I-522
`
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`Structures of reference compounds:
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`(1) Mevinolin
`
`(2) CS-514
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`
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`Table 3: Inhibitory activities by Test B-1
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`
`_T(L5
`/1‘Is5QO(molar concentration)
`v
`Compound
`
`
`(Compound
`of the present
`invention)
`
`I-5l
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`(Reference
`compound)
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`cs—514
`
`1
`
`x 1o'7
`
`3.5 x 1o‘7
`
`5
`
`l0
`
`15
`
`____________________________________________________
`
`20
`
`In Table 3-2,
`
`the relative activities are shown based
`
`on the activities of CS-514 being evaluated to be 1.
`
`25
`
`30
`
`35
`
`Table 3-2: Relative activities by Test B-1
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`‘"——'—-_-_—_"'__‘_‘_—_‘"_‘_—___“'_"—_‘—’_—_-_‘___-—‘_5‘
`Compound
`Relative activities
`'
`_______________________________________._______________
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`I-116
`
`I-520
`
`II-20
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`.
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`19.4
`
`20.0
`
`20.8
`
`A
`
`____________________________________________l________
`Results of the measurement of the inhibitor
`_____________________________________________;Z
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`activities by Test C
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`40
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`The percent decrease of counts after the oral
`
`administration of 0.05 mg/kg of compound I-520 was 55%
`
`relative to the measured value of the control group.
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`The
`
`percent decrease of counts after the oral administration
`
`of 10 mg/kg of CS-514 was 55% under the same condition.
`
`45 The compounds of the present
`
`invention exhibited
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`activities superior to the reference compound such as
`CS-514 or Mevinolin in Test A, and exhibited activities
`
`superior to CS-514 in Tests B and C.
`
`Test D: Acute toxicity
`
`A 0.5% CMC suspension of a test compound was orally
`
`administered to ICR male mice (group of three mice).
`
`The
`
`acute toxicity was determined based on the