Case 1:14-cv-02758-PAC Document 62-1 Filed 05/08/15 Page 1 of 19
`
`Kowa Company, Ltd. et al. v. Aurobindo Pharma Limited et al.,
`Civil Action No. 14-CV-2497 (PAC) (and related cases)
`
`
`
`
`
`Exhibit 1 to Declaration of Thomas R. Burns,
`dated May 8, 2015, in support of Defendants’
`Joint Opening Claim Construction Brief
`
`

`

`Case 1:14-cv-02758-PAC Document 62-1 Filed 05/08/15 Page 2 of 19
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`
`
`@‘.
`
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`UNITED STATES DEPARTMENT OF COMMERCE
`
`United States Patent and Trademark Office
`
`
`
`THIS IS TO CERTIFY THAT ANNEXED HERETO IS A TRUE COPY FROM
`
`THE RECORDS OF THIS OFFICE OF:
`
`March 28, 2014
`
`
`
`
`
`By Authority of the
`
`Under Secretary of Commerce for Intellectual Property
`and Director of the United States Patent and Trademark Office
`
`_
`
`Rowsmce
`
`T. LAWRENCE
`
`Certifying Officer
`
`
`
`
`
` U.S. PATENT: 5,856,336
`ISSUE DATE:
`January 05, I999
`
`
`
`
`
`
`
`
`
`MYLAN Pitav 009113
`
`

`

`Case 1:14-cv-02758-PAC Document 62-1 Filed 05/08/15 Page 3 of 19
`Case 1:14-cv-02758-PAC Document 62-1 Filed 05/08/15 Page 3 of 19
`
`U8005856336A
`5,856,336
`[11] Patent Number:
`[19]
`United States Patent
`
`Fujikawa et al. Jan. 5, 1999 [45] Date of Patent:
`
`
`[54] QUINOLINE TYPE MEVADONOLACTONES
`
`[75]
`
`Inventors: Yoshihiro Fujikawa; Mikio Suzuki;
`Hiroshi Iwasaki, all of Funabashi;
`Mitsuaki Sakashita; Masaki Kitahara,
`both of Shiraoka-machi, all of Japan
`
`[73} Assignee: Nissan Chemical Industries Ltd.,
`Tokyo, Japan
`[21] Appl. No.: 883,398
`
`[22] Filed:
`
`May 15,1992
`
`Related U.S. Application Data
`[62] Division of Ser. No. 631,092, Dec. 19, 1990, which is a
`continuation of Ser. No. 233,752, Aug. 19, 1988.
`
`Primary Examiner-Laura L. Stockton
`‘
`An
`A
`F'
`1
`gen; 0' “"013 on’ Splvak’
`“me”
`Mai“ 8‘ Neus‘ad" RC-
`
`M
`
`celeuand’
`
`[57]
`
`ABSTRACT
`
`A compound 0f the formula
`F
`
`[A]
`
`Z
`
`I
`
`[30]
`
`Foreign Application Priority Data
`
`N
`
`“'1”
`
`[JP]
`[JP]
`[JP]
`
`Aug. 20, 1987
`1311.26, 1988
`Aug. 3, 1988
`C 6
`
`63-15585
`Japan ................................ .. 62-207224
`Japan
`
`Japan
`63-193606
`5
`~
`1
`5
`"""""""" """" A6 K 31/2};
`Esg
`
`[58] Field oi‘ é£21?'I.............iiIIIIIZIIiI 546/1733 514/311
`’
`
`Z=_CH(OH)_CH2_CH(0H)_C1.12_COoyzca
`have HMG—CoA inhibiting eifects, making them use-
`ful as inhibitors of cholesterol biosynthesis. The com-
`Pound may be prepared as a pharmaceutical for reduc-
`ing hype'lipidemia' hyperlipwmteinemia °’
`atherosclerosis.
`
`[56]
`
`References Cited
`U.S. PATENT DOCUMENTS
`
`5,753,675
`
`5/1998 Wattanasin .............................. 514/311
`
`'
`
`2 Claims, No Drawings
`
`Copy provided by USPTO from the PIRS Image Database on 03/21/2014
`MYLAN(Pitav)009114
`
`

`

`Case 1:14-cv-02758-PAC Document 62-1 Filed 05/08/15 Page 4 of 19
`Case 1:14-cv-02758-PAC Document 62-1 Filed 05/08/15 Page 4 of 19
`
`1
`QUINOLINE TYPE MEVALONOLACTONES
`
`5,856,336
`
`10
`
`15
`
`20
`
`25
`
`30
`
`35
`
`40
`
`45
`
`50
`
`55
`
`60
`
`65
`
`This is a division, of application Ser. No. 07/631,092,
`filed on Dec. 19, 1990, which is a continuation of 07/233,
`752, filed Aug. 19, 1988.
`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.
`Some fermentation metabolic products such as
`compactine, CS-514, Mevinolin or semi-synthetic deriva-
`tives or fully synthetic derivatives thereof are known to be
`inhibitors against HMG-CoA reductase which is a rate
`limiting enzyme for cholesterol biosynthesis. (A. Endo J.
`Med Chem., 28(4) 401 (1985))
`CS-514 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 atherosclerosis.
`(IXth Int. Symp. Drugs Affect. Lipid Metab, 1986, p30,
`p31, p66)
`to fully synthetic derivatives,
`However, with respect
`particularly hetero aromatic derivatives of inhibitors against
`HMG—COA reductase, limited information is disclosed in the
`following literatures:
`WPI ACC NO. 84-158675, 86-028274, 86-098816,
`86-332070, 87-124519, 87—220987, 88-07781, 88-008460,
`88-091798 and 88-112505.
`The present inventors have found that mevalonolactone
`derivatives having a quinoline ring, the corresponding dihy-
`droxy 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 accomplished on the basis of this
`discovery.
`The novel mevalonolactone derivatives of the present
`invention are represented by the following formula 1:
`R3
`R4
`
`(1)
`
`Y— Z
`
`N
`
`R5
`
`R R
`
`2
`
`R1
`
`wherein R1, R2, R3, R4 and R6 are independently hydrogen,
`C“, alkyl, C3_6 cycloalkyl, C1_3 alkoxy, n-butoxy, i-butoxy,
`sec-butoxy, R7R8N— (wherein R7 and R8 are independently
`hydrogen or C1,3 alkyl), trifluoromethyl, trifluommethoxy,
`difluoromethoxy, fluoro, chloro, bromo, phenyl, phenoxy,
`benzyloxy, hydroxy,
`trimethylsilyloxy, diphenyl-t-
`butylsilyloxy, hydroxymethyl or ——O(CH2),OR19 (wherein
`R19 is hydrogen or CL3 alkyl, and 1 is 1, 2 or 3); or when
`located at the ortho position to each other, R1 and R2, or R3
`and R“ together form —CH=CH—CH=CH——; or when
`located at the ortho position to each other, R1 and R2
`together form —0C(R15)(R16)0—(wherein R15 and R16 are
`independently hydrogen or C1_3 alkyl); Y is —Cf12——,
`—CH,CH,—,
`-CH=CH——-, —CH2——CH=CH—~ or
`——CH=CH~—CH,—; and z is -Q-CH2WCH2-C02R12,
`
`R11
`HO
`
`2
`
`O
`
`O
`
`O
`
`11
`
`O
`
`0
`
`0
`
`R
`
`R17
`R18
`
`X
`0
`
`COZR12
`
`(wherein Q is —C(O)——, ——C(OR13)2—- or —CH(OH ;
`W is —C(O)——, —C(OR13)2— or —C(R“)(OH)———; R 1 is
`hydrogen or C1_3 alkyl; R12 is hydrogen or R14 (wherein R14
`is physiologically hydrolyzable alkyl or M (wherein M is
`NH4, sodium, potassium, 1/2 calcium or a hydrate of lower
`alkylamine, di-lower alkylamine or tri-lower alkylamine»;
`two R13 are independently primary or secondary C1_5 alkyl;
`or two R13 together form ——(CH 2— or —(CH2)3—; R17
`and R18 are independently hydrogen or CL3 alkyl; and R5 is
`hydrogen, C” alkyl, C2,; alkenyl, CM cycloalkyl,
`R9
`
`(wherein R9 is hydrogen, C1_4 alkyl, Cm alkoxy, fluoro,
`chloro, bromo or trifluoromethyl), phenyl-(CH2)m—
`(wherein m is 1, 2 or 3), —(CH2),,CH(CH3)-phenyl or
`phenyl-(CHQHCH(CH3)—— (wherein n is 0, 1 or 2).
`Various substituents in the formula I will be described in
`detail with reference to specific examples. However, it
`should be understood that the present invention is by no
`means restricted by such specific examples.
`CM; alkyl for R1, R2, R3, R4, R6 and R9 includes, for
`example, methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl,
`sec-butyl and t-butyl. C1_3 alkoxy for R1, R2, R3, R4 and R6
`includes, for example, methoxy, ethoxy, n-propoxy and
`i-propoxy.
`C1_3 alkyl [or R11 includes, for example, methyl, ethyl,
`n-propyl and i-propyl.
`CL3 alkyl for R13 includes, for example, methyl, ethyl,
`n-propyl and i—propyl.
`for example, methyl, ethyl,
`Alkyl
`for R14 includes,
`n-propyl, i-propyl, n-butyl and i-butyl.
`M is a metal capable of forming a pharmaceutically
`acceptable salt, and it includes, for example, sodium and
`potassium.
`CO2M includes, for example, —COZ.NH4 and —COZH.
`(primary to tertiary lower alkylamine such as
`trimethylamine).
`'
`CH, alkyl for R5 includes, for example, methyl, ethyl,
`n-propyl,
`i-propyl, n-butyl,
`i-butyl, sec-butyl,
`t—butyl,
`n-pentyl and n-hexyl.
`C3_6 cycloalkyl for Rs includes, for example, cyclopropyl,
`cyclobutyl, cyclopentyl and cyclohexyl.
`C2_3 alkcnyl for R5 includes, for example, vinyl and
`i-propenyl.
`Phenyl-(CH2)m- for R5 includes, for example, benzyl,
`B-phenylethyl and y-phenylpropyl.
`Phenyl—(CHZ),,CH(CH3)— for R5 includes, for example,
`a-phenylethyl and u-benzylethyl.
`Cm alkyl for R7 and R8 includes, for example, methyl,
`ethyl, n-propyl and i-propyl.
`Further, these compoundsmay have at least one or two
`asymmetric carbon atoms and may have at least two to four
`
`Copy provided by USPTO from the PIRS Image Database on 03/21/2014
`MYLAN(Pitav)009115
`
`

`

`Case 1:14-cv-02758-PAC Document 62-1 Filed 05/08/15 Page 5 of 19
`Case 1:14-cv-02758-PAC Document 62-1 Filed 05/08/15 Page 5 of 19
`
`5,856,336
`
`3
`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 fall-
`ing outside the definition of ——COZR12 of the carboxylic
`acid moiety of substituent Z of the compounds of the present
`invention, those which undergo physiological hydrolysis,
`after intake, to produce the corres onding carboxylic acids
`(compounds wherein the ~—C02R 2 moiety is ~——C02H) are
`equivalent to the compounds of the present invention.
`Now, preferred substituents of the compounds of the
`present invention will be described.
`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. 1' 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.
`Preferred substituents for R1, R2 and R6 are hydrogen,
`fluoro, chloro, bromo, CL3 alkyl, CL3 alkoxy, C3_6
`cycloalkyl, dimethylamino, hydroxy, hydroxymethyl,
`hydroxyethyl,
`trifluoromethyl,
`trifluoromethoxy,
`difluoromethoxy, phenoxy and benzyloxy.
`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‘~methy1,
`4'-methyl, 4'-chloro and 4'-fluoro.
`Other preferred combinations of R3 and R4 include
`3'-methy1-4'-chloro, 3',5'-dichloro, 3',5‘-difiuoro, 3',5‘-
`dimethyl and 3’-methyl-4'-fluoro.
`Preferred examples for R5 include primary and secondary
`CL6 alkyl and C3_6 cycloalkyl.
`Preferred examples for Y include —CH2—CH2—— and
`—CH=CH-—.
`Preferred examples for Z include
`HO
`O
`O
`
`——CH(0H)CH2CH2(0H)CH,C02R12, ~CH(OH)CH2C(O)
`CH2c02R12 and ——-CH(OH)CH2C(OR13)2CH2C02R12.
`Now, more preferred substituents of the compounds of the
`present invention will be described.
`As more preferred examples for R1, R2 and R5, when both
`R2 and R6 are hydrogen, R1 is hydrogen, 5~fluoro, 6-fluoro,
`7-fluoro, 8-fluoro, 5-chloro, 6-chloro, 7-chloro, 8-chloro,
`5-bromo, 6-bromo, 7-bromo, 8-bromo, S-methyl, 6-methyl,
`7-methy1, 8-methyl, S-methoxy, 6-methoxy, 7-methoxy,
`8-methoxy, 5-trifluoromethyl, 6-trifluoromethyl,
`7-trifluoromethyl, 8-trifluoromethyl, 6-trifluoromethoxy,
`6-difluoromethoxy, 8-hydroxyethyl, 5-hydroxy, 6-hydroxy,
`7-hydroxy, 8-hydroxy, 6-ethyl, 6-n-buty1 and
`7-dimethylamino.
`When R5 is hydrogen, R1 and R2 together represent
`6-chloro-8-methyl, 6—bromo-7-methoxy, 6-methy1-7-chlor0,
`6-chloro-8-hydroxy, 5-methyl-2-hydroxy, 6-methoxy-7-
`chloro, 6-chloro-7-methoxy, 6-hydroxy-7—chloro, 6-chloro-
`7-hydroxy, 6-chloro-8-bromo, 5-chloro-6-hydr0xy,'
`6-bromo-8-chloro, 6-bromo-8-hydroxy, 5-methyl-8-chloro,
`7-hydroxy-8-chloro, 6-bromo-8-hydroxy, 6-methoxy-7-
`methyl, 6-chloro-8-bromo, 6-methyl-8-bromo, 6,7-difluoro,
`6,8-difluoro, 6,7-methy1enedioxy, 6,8-dichloro, 5,8-
`
`4
`dimethyl, 6,8-dimethyl, 6,7-dimethoxy, 6,7-diethoxy, 6,7-
`dibromo or 6,8-dibromo.
`When R1, R2 and R6 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 R3 and R4, when R3 is
`hydrogen, R4 is hydrogen, 4'-methyl, 4'—chloro or 4'-fluoro.
`When both R3 and R4 are not hydrogen,
`they together
`represent 3',5'-dimethyl or 3'-methyl~4'-fluoro.
`As more preferred examples for R5, the above-mentioned
`preferred examples of R5 may be mentioned.
`As preferred examples for Y, —CH2———CH2-— and (E)—
`——-CH=CH—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 examples for
`R1, R2 and R6 , when both R2 and R‘5 are hydrogen, R1 is
`hydrogen, 6-methy1, 6-ethyl, 6-trifluoromethyl, 6-hydroxy,
`G-methoxy, 6-chloro, 6-bromo, 6-n-buty1 and
`7-dimethylamino.
`When only R6 is hydrogen, R1 and R2 represent 6,8-
`dichloro, 5,8-dimethyl, 6,8-dimethyl, 6,7-dimethoxy, 6,7-
`diethoxy, 6,7—dibromo, 6,8-dibromo, 6,7-difluoro and 6,8-
`difluoro.
`As still further preferred examples for R3 and R4, when R3
`is hydrogen, R4 is hydrogen, 4'-chloro or 4'-fluoro, or R3 and
`R4 together represent 3’-methyl-4'-fluoro.
`Still further preferred examples for R5 include ethyl,
`n-propyl, i-propyl and cyclopropyl.
`Still further preferred examples for Y include (E)—
`CH=CH—.
`
`the above-
`As still further preferred examples for Z,
`mentioned preferred example for Z may be mentioned.
`Now, the most preferred substituents for the compounds
`of the present invention will be described.
`As the most preferred examples for R], R2 and R6, when
`both R2 and R6 are hydrogen, R1 is hydrogen, 6-methyl or
`6-chloro.
`
`When only R6 is hydrogen, R1 and R2 together represent,
`for example, 6,7-dimethoxy.
`As the most preferred examples for R3 and R4, R3 is
`hydrogen and R4 is hydrogen, 4'-chloro or 4'-fluoro.
`The most preferred examples for Rs include i~propy1 and
`cyclopropyl. The most preferred example for Y may be
`(E)——-CH=CH—.
`the above-
`As the most preferred examples for Z,
`mentioned preferred examples for Z may be mentioned.
`Now, particularly preferred specific compounds of the
`present invention will be presented. The following com-
`pounds (a) to (z) are shown in the form of carboxylic acids.
`However, the present invention include not only the com-
`pounds in the form of carboxylic acids but also the corre-
`sponding lactones formed by the condensation of the car-
`boxylic acids with hydroxy at the 5-position, and sodium
`salts and lower alkyl esters (such as methyl, ethyl, i—propyl
`and n-propyl esters) of the carboxylic acids, which can be
`physiologically hydrolyzed to the carboxylic acids.
`(a)
`(E)-3,5-dihydroxy-7-[4'-(4"-fluorophenyl)-2'—(1"-
`methylethyl)-quinolin—3‘-yl]-hept-6-enoic acid
`(b)
`(E)-3,5-dihydroxy-7-[4'-(4"-fluorophenyl)—2‘-(1“-
`methylethyl)—6'-chloro-quinolin-3'-y1]-hept-6-enoic acid
`(c)
`(E)—3,5-dihydroxy-7-[4'-(4" -fluorophenyl)-2'-(1"-
`methylethyl)—6‘-methy1-quinolin-3'-yl]-hept-6-enoic acid
`(d)
`(E)-3,5-dihydroxy-7-[4'-(4"-fluoropheny1)-2'-
`(1"methylethyl)-6',7'-dimethoxy-quinolin-3'-yl]—hept-6-
`enoic acid
`
`5
`
`10
`
`15
`
`20
`
`25
`
`30
`
`35
`
`40
`
`45
`
`50
`
`55
`
`60
`
`65
`
`Copy provided by USPTO from the PIRS Image Database on 03/21/2014
`MYLAN(Pitav)009116
`
`

`

`Case 1:14-cv-02758-PAC Document 62-1 Filed 05/08/15 Page 6 of 19
`Case 1:14-cv-02758-PAC Document 62-1 Filed 05/08/15 Page 6 of 19
`
`5,856,336
`
`5
`
`5
`(e) (E) -3, 5 -dihydroxy-7-[4'-(4" -fluo rophenyl)-2 '-
`cyclopropyl-quinolin-3'~yl]-hept-6-enoic acid
`(f)
`(E)-3,5 -dihydroxy-7-[4'-(4"-fluor0phenyl)-2'-
`cyclopropyl-6'-chloro-quinolin-3'-y1]-hept-6-enoic acid
`(g)
`(E)-3,5~dihydroxy-7-[4'—(4" -fluorophenyl)-2'-
`cyclopropyl~6'-methyl-quinolin-3’-yl]-hept-6-enoic acid
`(h)
`(E)-3,5-dihydroxy~7-[4'—(4" —fluorophenyl)—2'—
`cyclopropyl-6',7'-dimethoxy-quino1in-3'-yl]-hept-6-en0ic
`acid
`(i) (E)-3,5-dihydroxy-7-[4'—(4"~chloropheny])-2'-(1"- 10
`methylethyl)-quinolin-3‘-yl]-l1ept-6-encic acid
`(E)-3,5-dibydroxy-7—[4'-(4"-chloropheny1)-2'-(1"—
`methylethyl)-6‘-chloro-quinolin-3'-y1]-hept-6-enoic acid
`(k) (E)-3,5~dihydroxy-7-[4'-(4"-chloropheny])—2'-(1"-
`methylethyl)~6'—methyl-quinolin-3‘-y1]-hcpt-6-enoic acid
`(l) (E)-3,5-dihydroxy-7-[4’-(4"-chlorophenyl)-2‘~(1"-
`methylethyl)-6',7'-dimethoxy-quinolin—3'-yl]-hept-6-enoic
`acid
`(E)-3,5-dihydroxy-7-[4'-(4"-chlorophenyl)-2’-
`(m)
`cyclopropyl-quinolin-S'-yl]—hept-6-enoic acid
`([1) (E)-3,5-dihydroxy-7-[4'-(4"-chloropheny1)-2'-
`cyclopropyl-é'-chloro-quinolin-3'-y1]-hept—6-enoic acid
`(0) (E)-3,5-dihydroxy-7-[4'-(4"-chlorophenyl)—2'-
`cyclopropy1-6'-methyI-quinolin-S‘-y1]-hept-6-enoic acid
`(p) (E)-3,5~dihydroxy-7—[4'-(4"—chloropheny1)-2'- 25
`cyclopropyl-6'7’-dimeth0xy-quinolin-3'-yl]-hept—6-enoic
`acid
`
`15
`
`20
`
`(q) (E)-3,S-dihydroxy-7-[4'-pheny1—2'—(1"-methylethyl)-
`quinolin-3'-yl]-hept-6-enoic acid
`(I) (E)-3,5—dihydroxy-7—[4‘-phenyl—2'-(1"-methylcthyl)— 30
`6'-chloro-quinolin-3'-yl]-hepl-6-enoic acid
`(5)
`(E)-3,5-dihydroxy—7-[4'—phenyl—2‘-(1"-methylethyl)-
`6'-methyl~quinolin-3'—y1]—hept-6-enoic acid
`(t) (E)-3,5-dihydroxy-7-[4'-phenyl-2’-( 1"-methylethy1)-
`6‘,7‘-dimethoxy—quinolin-B'-yl]—hept-6-enoic acid
`(u) (E)-3,5-dihydroxy-7-[4'-pheny1-2'-cyclopropyl-
`quinolin-3'-y1]-hept-6-enoic acid
`(v) (E)-3,5-dihydroxy-7-[4'-pheny1—2'-cyelopropy1-6'-
`chloro-quinolin-3’-yl]-hept-6-enoic acid
`(w)
`(E)-3,5-dihydroxy-7-[4'-phenyl-2’~cyclopropy1—6'- 40
`methyl-quinolin-3'-yl]-hepl-6-enoic acid
`(x) (E)-3,5-dihydroxy-7-[4‘-phenyl-2‘-cyclopropy1—6',7'-
`dimethoxy-quinolin-B'-yl]-hcpt-6-enoic acid
`(y) (E)-3,5-dihydroxy-7—[4'-(4"-fluorophenyl)-2'-(1"-
`methylethyl)—6'~methoxy-quinolin-3‘~yl]-hept-6-enoic acid
`(z) (E)-3,5-dihydroxy-7-[4'—(4"-fluorophenyl)-2'-
`cyclopropy1-6'-methoxy-quinolin—3’-y1]-hept-6-enoic acid
`The mevalouolactones of the formula I can be prepared by
`the following reaction scheme. The enal 111 can also be
`prepared by processes K, L and M.
`R3
`R4
`
`35
`
`45
`
`50
`
`6
`-30ntmued
`R4
`
`R3
`
`CH OH
`2
`
`R5
`
`O
`N
`VI
`
`R3
`
`R4
`
`CH0
`
`R5
`
`N
`
`v
`
`R3
`
`R4
`
`/ OE:
`
`H
`
`0 °
`R
`N
`5
`IV
`
`, R3
`
`R4
`
`CH0
`
`N
`
`m
`
`R5
`
`R6
`R1
`
`2
`
`R
`
`R
`R:
`
`R
`
`‘
`
`R5
`R2
`
`l
`
`R
`
`Ra
`R2
`
`R1
`
`_>
`B
`
`79
`
`D>
`
`T9
`
`COZRIZ
`
`55
`
`Copy provided by USPTO from the PIHS Image Database on 03/21/2014
`MYLAN(Pitav)009117
`
`

`

`Case 1:14-cv-02758-PAC Document 62-1 Filed 05/08/15 Page 7 of 19
`Case 1:14-cv-02758-PAC Document 62-1 Filed 05/08/15 Page 7 of 19
`
`5,856,336
`
`7
`-continued
`R4
`
`R3
`
`0H
`
`8
`-continued
`R4
`
`R3
`
` fi
`
`10
`
`15
`
`20
`
`25
`
`30
`
`35
`
`R6
`
`R2
`
`1
`
`R
`
`R6
`
`R2
`
`1
`
`R
`
`CHzOH
`
`fi
`M
`
`0
`N
`
`R5
`
`R3
`
`[X
`
`R4
`
`I
`
`l
`
`CHO
`
`R5
`
`N
`
`III
`
`R3
`
`R4
`
`OH
`
`COZR12
`
`0H
`
`0
`
`fl
`
`R6
`
`R2
`
`com”
`
` l
`
`R
`
`1-1
`
`R4
`
`R3
`
`OH
`
`.
`In the above reacllon scheme, R1, R2, R3, R4, R5, R6 and
`R12 are as defined above with respect to the formula I, and
`
`40
`
`45
`
`50
`
`55
`
`60
`
`65
`
`K
`
`N
`
`R5
`
`R1
`
`[-4
`R4
`
`R3
`
`R6
`R2
`
`1
`
`R
`
`CHO
`
`R5
`
`O
`N
`
`V
`
`‘
`Copy provided by USPTO from the PIRS Image Database on 03/21/2014
`MYLAN(Pitav)009118
`
`

`

`Case 1:14-cv-02758-PAC Document 62-1 Filed 05/08/15 Page 8 of 19
`Case 1:14-cv-02758-PAC Document 62-1 Filed 05/08/15 Page 8 of 19
`
`5,856,336
`
`9
`R21 and R22 independently represent C1,4 lower alkyl such
`as methyl, ethyl, n-propyl, i-propyl or n-butyl.
`,
`Step A represents a reduction reaction of the ester to a
`primary alcohol. Such reduction reaction can be conducted
`by using various metal hydrides, preferably diisobutylalu-
`minium hydride, in a solvent such as tetrahydrofuran or
`toluene at a temperature of from —20° to 20° C., preferably
`from —-10° to 10° C.
`
`Step B represents an oxidation reaction of the primary
`alcohol to an aldehyde, which can be conducted by using
`various oxidizing agents. Preferably, the reaction can be
`conducted by using pyridinium chlorochromate in methyl-
`ene chloride at a temperature of from 0° to 25 ° C., or by
`using oxalyl chloride, dimethyl sulfoxide and a tertiary
`amine such as triethylamine (Swern oxidation), or by using
`a sulfur trioxide pyridine complex.
`Step C represents a synthesis of a 3-ethoxy-1-hydroxy-
`2-propene derivative, which can be prepared by reacting a
`compound V to lithium compound which has been prelimi-
`narily formed by treating cis-1-ethoxy-2—(tri-n-butylstanny1)
`ethylene with butyl lithium in tetrahydrofuran.
`As the reaction temperature, it is preferred to employ a
`low temperature at a level of from —60° to —78° C.
`Step D represents a synthesis of an enal by acidic hydroly-
`sis. As the acid catalyst, it is preferred to employ p—toluene
`sulfonic acid, hydrochloric acid or sulfuric acid, and the
`reaction may be conducted in a solvent mixture of water and
`tetrahydrofuran or ethanol at a temperature of from 10° to
`25° C. The 3-ethoxy-1-hydroxy-2-propene derivative
`obtained in Step C can be used in Step D without purification
`i.e. by simply removing tetra-n-butyl tin formed simulta-
`neously.
`Step E represents a double anion condensation reaction
`between the enal III and an acetoacetate. Such condensation
`reaction is preferably conducted by using sodium hydride
`and n-butyl lithium as the base in tetrahydrofuran at a
`temperature of from —80° to 0" C., preferably from —30° to
`-10° C.
`
`Step F represents a reduction reaction of the carbonyl
`group, which can be conudcted by using a metal hydride,
`preferably sodium borohydride in ethanol at a temperature
`of from —10° to 25° 0, preferably from —10° to 5° C.
`Further, the reduction reaction may be conducted by using
`zinc borohydride in dry ethyl ether or dry tetrahydrofuran at
`a temperature of —100° to 25° C., preferably from —80° to
`—50° C.
`
`Step G is a step for hydrolyzing the ester. The hydrolysis
`can be conducted by using an equimolar amount of a base,
`preferably potassium hydroxide or sodium hydroxide, in a
`solvent mixture of water and methanol or 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. The
`dehydration reaction can be conducted in benzene or toluene
`under reflux while removing the resulting water or by adding
`a suitable dehydrating agent such as molecular sieve.
`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 N-cyclohexyl-N'-[2'-(methylmorpholinium)ethy1]
`carbodiimide 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 quino-
`line ring. This hydrogenation reaction can be conducted by
`using a catalytic amount of palladium—carbon or rhodium—
`
`10
`
`15
`
`20
`
`25
`
`30
`
`35
`
`4o
`
`45
`
`50
`
`55
`
`60
`
`65
`
`10
`carbon in a solvent such as methanol, ethanol, tetrahydro—
`furan or acetonitrile at a temperature of from 0° to 50° C.,
`preferably from 10° to 25° C.
`Step K represents a reaction for the synthesis of an
`a,[3-unsaturated carboxylic acid ester, whereby a trans-form
`0t,B-unsaturated carboxylic acid ester can be obtained by a
`so-called Homer-Wittig reaction by using an alkoxycarbo-
`nylmethyl 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 ~15° C.
`Step L represents a reduction reaction of the (1,6-
`unsaturated carboxylic acid ester to an ally] alcohol. This
`reduction reaction can be conducted by using various metal
`hydrides, preferably diisobutylaluminiumhydride, in a sol-
`vent such as dry tetrahydrofuran or toluene at a temperature
`of from ~10" to 10° C., preferably fi'om -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 active man-
`ganese dioxide,
`in a solvent such as tetrahydrofuran,
`acetone, ethyl ether or ethyl acetate at a temperatrue of from
`0° to 100° C., preferably from 15° to 50° C.
`Step N represents a reaction for the synthesis of an
`ot,B-unsaturated ketone by the selective oxidation of the
`dihydroxy carboxylic acid ester. This reaction can be con-
`ducted by using activated manganese dioxide in a solvent
`such as ethyl ether, tetrahydrofuran, benzene or toluene at a
`temperature of from 20° to 80° C., preferably from 40° to
`80° C.
`
`In addition to the compounds disclosed in Examples given
`hereinafter, compounds of the formulas [-2 and 1-5 given in
`Table 1 can be prepared by the process of the present
`invention. In Table 1, 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.
`
`TABLE 1
`
`R“
`
`OH
`
`1-2 (R12= H)
`H (R12==Na)
`
`
`
`
`
`
`
` R‘ R2 R3 R4 R5 R6
`
`6-0Me
`H
`H
`H
`i-Pr
`H
`6-0Me
`H
`4-F
`H
`i-Pr
`H
`6-Br
`H
`4—F
`H
`i-Pr
`H
`6-Me
`S-Me
`4-F
`H
`i—Pr
`H
`7-0Me
`8-0Me
`4-F
`H
`i-Pr
`H
`6-Br
`H
`Z-F
`H
`i—Pr
`H
`
`6,7
`
`4—F
`
`H
`
`i-Pr
`
`H
`
`Copy provided by USPTO from the PIRS Image Database on 03/21/2014
`MYLAN(Pitav)009119
`
`

`

`Case 1:14-cv-02758-PAC Document 62-1 Filed 05/08/15 Page 9 of 19
`Case 1:14-cv-02758-PAC Document 62-1 Filed 05/08/15 Page 9 of 19
`
`5,856,336
`
`11
`
`TABLE 1-continued
`
`
`R3
`
`R‘l
`
`OH
`
`1—2 (R12 - II)
`1‘5 (R12 -= Na)
`
`COZRIZ
`
`OH
`
`R6
`R2
`
`R1
`
`I
`
`N
`
`R5
`
`R1
`R2
`R3
`R4
`V R5
`R6
`
`H
`H
`4~F
`H
`H
`
`H
`H
`6-Cl
`6—Cl
`6-0CH2Ph
`H
`H
`6-Cl
`6-Me2N
`6-Me
`6—i—Pr
`7-Me
`6-0Me
`6-Br
`6-i-Pr
`6-C1
`5-F
`6—OMe
`6-Me
`6-Cl
`H
`H
`6-OMe
`6-0Me
`6-0Me
`6-0Me
`6-OMe
`6-Me
`6-Me
`6-Me
`G-Me
`6-Me
`6-Cl
`6-Cl
`6-Cl
`6—C1
`6-C1
`H
`H
`H
`H
`H
`
`H
`H
`H
`H
`H
`H
`H
`H
`H
`[I
`H
`H
`H
`H
`H
`8-Cl
`6-Br
`7-0Me
`7-Me
`7-Cl
`H
`H
`7-0Me
`7-0Me
`7-0Me
`7-0Me
`7-0Me
`H
`H
`H
`H
`H
`H
`II
`H
`H
`H
`H
`H
`H
`H
`H
`
`4-Ph
`4-l’hCH2
`4-F
`4-F
`4-F
`4-F
`4-F
`4-F
`4-F
`4—F
`4-F
`4-F
`4-F
`4-F
`4-F
`4—F
`4-F
`4-F
`4-F
`4F
`4-F
`4—F
`H
`4—C1
`H
`4-Cl
`4-F
`H
`4-Cl
`H
`4-C1
`4‘F
`H
`4-Cl
`H
`4-Cl
`4»F
`H
`4-Cl
`H
`4-Cl
`4-F
`
`H
`H
`H
`H
`H
`H
`H
`H
`H
`H
`H
`H
`H
`H
`H
`H
`H
`H
`H
`H
`H
`H
`H
`H
`H
`H
`H
`H
`H
`H
`H
`H
`H
`H
`H
`H
`H
`H
`H
`H
`H
`H
`
`i-l’r
`i—Pr
`c-Pr
`sec-Bu
`i-Pr
`i-Bu
`c-Pent
`c-Pent
`i~l’r
`c—Pr
`i-Pr
`c-Pt'
`c-Pr
`c—Pr
`c—Pr
`c-Pr
`i-Pr
`i-l’r
`i-l’r
`i—Pr
`c-Bu
`c-Hex
`i-Pr
`i-Pr
`c-I’r
`c-l’r
`c—Pr
`i-Pr
`i-Pr
`c-l’r
`c-Pr
`c-l’r
`i-Pr
`i-Pr
`o—Pr
`c—Pr
`c—Pr
`i—Pr
`i-Pr
`c-Pr
`c-Pr
`c-Pr
`
`H
`H
`H
`H
`H
`H
`H
`H
`H
`H
`H
`H
`H
`H
`H
`H
`8-Br
`8-0Me
`8-Me
`8-Cl
`H
`H
`H
`H
`H
`H
`H
`H
`H
`H
`H
`H
`H
`H
`H
`H '
`H
`H
`H
`H
`H
`
`12
`They may be formulated into various suitable formula-
`tions 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 form of physiologically
`hydrolyzable and acceptable esters or lactones, or pharma-
`ceutically acceptable salts.
`The pharmaceutical composition of the present invention
`is preferably administered orally in the form of the com-
`pound 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 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 suppository formulated
`by using oily base material such as cacao butter, polyethyl-
`ene glycol, lanolin or fatty acid 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 formula-
`tion 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 gas-
`trointestinal tract.
`
`The daily dose of the compound of the formula I is 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 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 1. Accordingly, the present inven-
`tion relates also to the compounds of the formulas II 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 pharmaco-
`logical activities of the compounds of the present invention,
`their Preparation Examples and Formulation 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 acetate in
`vitro
`
`Enzyme solution was prepared from liver of male Wistar
`rat billialy cannulated and discharged bile forever 24 hours.
`Liver was cut out at mid-dark and microsome and supema-
`tant fraction which was precipitable with 40-80% of satu—
`ration of ammonium sulfate (sup fraction) were prepared
`from liver homogenate according to the modified method of
`Knauss et. al.; Kuroda, M., et. al., Biochim. Biophys. Acta,
`489, 119 (1977). 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 [41 of the
`
`10
`
`15
`
`20
`
`25
`
`30
`
`35
`
`4o
`
`45
`
`50
`
`55
`
`60
`
`65
`
`Hl F
`
`urther, pharmaceutically acceptable salts such as potas-
`sium 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 cholesterol
`in blood as lipoprotein. Thus, the compounds of the present
`invention are useful as curing agents against hyperlipidemia,
`hyperlipoproteinemia and atheroscleosis.
`
`Copy provided by USPTO from the PIRS Image Database on 03/21/2014
`MYLAN(Pitav)009120
`
`

`

`Case 1:14-cv-02758-PAC Document 62-1 Filed 05/08/15 Page 10 of 19
`Case 1:14-ov-02758-PAC Document 62-1 Filed 05/08/15 Page 10 of 19
`
`5,856,336
`
`14
`
`TABLE 2
`Inhibitog activities by Tbst A
`
`Compound I50 (molar concentration)
`
`(Compounds of the
`present invention)
`[-13
`[-51
`1-52
`1-53
`(Reference compounds)
`1.4 x 10‘8
`Mevinolin
`
`cs-514 9.0 x 10-9
`
`1.25 x 10‘7
`1.0 X 10—8
`7.1 X 10—8
`1.9 x 1077
`
`10
`
`15
`
`In Table 2-2, the relative activities are shown based on the
`activities of CS-514 being evaluated to be 1.
`
`20
`
`TABLE 2-2
`Relative activities by Test A
`
`
`Compound Relative activities
`(Compounds of the
`present invention)
`1.75
`[-16
`2.25
`[-116
`0.37
`I—117
`3.21
`I-120
`
`I-522 0.76
`
`Structures of reference compounds:
`
`(1) Mevinolin
`
`
`
`(2) 03-514
`
`Na02C
`
`OH
`
`13
`reaction mixture containing ATP; 1 mM, Glutathione; 6 mM,
`Glucose-l-phosphate; 10 mM, NAD; 0.25 mM, NADP; 0.25
`mM, CoA; 0.04 mM and 0.2 mM [2-14C]sodium acetate (0.2
`ad) with 4 ,ul of test compound solution dissolved in water
`or dimethyl sulfoxide. To stop reaction and saponify, 1 m1 of
`15% EtOH-KOH was added to the reactions and heated at
`75° C. for 1 hour. Nonsaponifiable lipids were extracted with
`petroleum ether and incorporated l4C radioactivity was
`counted. Inhibitory activity of compounds was indicated
`with IC50.
`
`Test B: Inhibition of cholesterol biosynthesis in culture cells
`Hep G2 cells at over 5th passage were seeded to 12 well
`plates and incubated with Dulbecco’s modified Eagle
`(DME) medium containing 10% of fetal bovine serum
`(FBS) at 37° C., 5% CO2 until cells were confluent for about
`7 days. Cells were exposed to the DME medium containing
`5% of lipoprotein deficient serum (LpDS) prepared by
`ultracentrifugation method for over 24 hours. Medium was
`changed to 0.5 m1 of fresh 5% LpDS containing DME before
`assay and 10 ,ul of test compound solution dissolved in water
`or DMSO were added. 0.2 uCi of [2-14C]sodium acetate (20
`,ul) was added at O hr(B-l) or 4 hrs(B-2) after addition of
`compounds. After 4 hrs further incubation with [2-14C]
`sodium acetate, medium was removed and cells were
`washed with phosphate buffered saline(PBS) chilled at 4° C.
`Cells were scraped with rubber policeman and collected to
`tubes with PBS and digested with 0.2 m1 of 0.5N KOH at
`37° C. Aliquot of digestion was used for protein analysis and
`remaining was saponified with 1 ml of 15% EtOH-KOH at
`75° C. for 1 hour. Nonsaponifiable lipids were extracted with
`petroleum ether and 14C radioactivity was counted. Counts
`were revised by cell protein and indicated with DPM/mg
`protein. Inhibito