United States Patent
`
`[19]
`
`[11] Patent Number:
`
`5,856,336
`
`Fujikawa et al.
`
`[45] Date of Patent:
`
`Jan. 5, 1999
`
`US005856336A
`
`[54] QUINOLINE TYPE MEVALONOLACTONES
`
`[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.
`
`[30]
`
`Foreign Application Priority Data
`
`Aug. 20, 1987
`Jan. 26, 1988
`Aug. 3, 1988
`
`[JP]
`[JP]
`[JP]
`
`Japan ................................ .. 62—207224
`.... .. 63-15585
`Japan
`
`Japan ................................ .. 63—193606
`
`Int. Cl.5 ....................... .. A61K 31/47; C07D 215/12
`[51]
`[52] U.S. Cl.
`........................................... .. 514/311; 546/173
`[58] Field of Search ............................ .. 546/173; 514/311
`
`[56]
`
`References Cited
`U.S. PATENT DOCUMENTS
`
`Primary Examiner—Laura L. Stockton
`Attorney, Agent, or Firm—Oblon, Spivak, McClelland,
`Maier & Neustadt, P.C.
`
`[57]
`
`ABSTRACT
`
`A compound of the formula
`
`[A]
`
`
`
`Z=—CH(OH)—CH2—CH(OH)—CH2—COO.%Ca
`have HMG—CoA inhibiting effects, making them use-
`ful as inhibitors of cholesterol biosynthesis. The com-
`pound may be prepared as a pharmaceutical for reduc-
`ing hyperlipidemia, hyperlipoproteinemia or
`atherosclerosis.
`
`5,753,675
`
`5/1998 Wattanasin ............................ .. 514/311
`
`2 Claims, No Drawings
`
`Mylan Exhibit 1001, Page 1
`
`
`
`Mylan Exhibit 1001, Page 1
`
`

`
`5,856,336
`
`1
`QUINOLINE TYPE MEVALONOLACTONES
`
`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 I:
`
`R3
`
`R4
`
`(1)
`
`R5
`R2
`
`R1
`
`Y— Z
`
`N
`
`R5
`
`wherein R1, R2, R3, R4 and R6 are independently hydrogen,
`C1_6 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, trifluoromethoxy,
`difluoromethoxy, fluoro, chloro, bromo, phenyl, phenoxy,
`benzyloxy, hydroxy,
`trimethylsilyloxy, diphenyl-t-
`butylsilyloxy, hydroxymethyl or —O(CH2),OR19 (wherein
`R19 is hydrogen or C1_3 alkyl, and l is 1, 2 or 3); or when
`located at the ortho position to each other, R1 and R2, or R3
`and R4 together form —CH=CH—CH=CH—; or when
`located at
`the ortho position to each other, R1 and R2
`together form —OC(R13)(R16)O—(wherein R13 and R16 are
`independently hydrogen or C1_3 alkyl); Y is —CH2—,
`—CH2CH2
`,
`CH—CH ,
`CH2 CH—CH
`or
`—CH=CH—CH2—; and Z is -Q-CHZWCHZ-CO2R12,
`
`10
`
`15
`
`20
`
`25
`
`30
`
`35
`
`40
`
`45
`
`50
`
`55
`
`60
`
`65
`
`R11
`HO
`
`2
`
`O
`%
`
`O
`
`0
`
`O
`
`0
`
`0
`
`R11
`
`R17
`1118*
`0
`
`CO R12
`2
`
`(wherein Q is —C(O)—, —C(OR13)2— or —CH(OH)—;
`W is —C(O)—, —C(OR13 2— or —C(R11)(OH)—; R11 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_6 alkyl;
`or two R13 together form —(CH2)2— or —(CH2)3—; R17
`and R18 are independently hydrogen or C1_3 alkyl; and R3 is
`hydrogen, C1_6 alkyl, C2_3 alkenyl, C3_6 cycloalkyl,
`
`R9
`
`(wherein R9 is hydrogen, C1_4 alkyl, C1_3 alkoxy, fluoro,
`chloro, bromo or
`trifluoromethyl), phenyl-(CH2)m—
`(wherein m is 1, 2 or 3), —(CH2)nCH(CH3)-phenyl or
`phenyl-(CH2)nCH(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.
`C1_6 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 for R11 includes, for example, methyl, ethyl,
`n-propyl and i-propyl.
`C1_3 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, —CO2NH4 and —CO2H.
`(primary to tertiary lower alkylamine such as
`trimethylamine).
`C1_6 alkyl for R3 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 R3 includes, for example, cyclopropyl,
`cyclobutyl, cyclopentyl and cyclohexyl.
`C2_3 alkenyl for R3 includes, for example, vinyl and
`i-propenyl.
`Phenyl-(CH2)m- for R3 includes, for example, benzyl,
`[3-phenylethyl and y-phenylpropyl.
`Phenyl-(CH2)nCH(CH3)— for R3 includes, for example,
`ot-phenylethyl and ot-benzylethyl.
`C1_3 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
`
`Mylan Exhibit 1001, Page 2
`
`Mylan Exhibit 1001, Page 2
`
`
`

`
`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 —CO2R12 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 corresponding carboxylic acids
`(compounds wherein the —CO2R12 moiety is —CO2H) 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, C1_3 alkyl, C1_3 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'-methyl,
`4'-methyl, 4‘-chloro and 4‘-fluoro.
`Other preferred combinations of R3 and R4 include
`3‘-methyl-4‘-chloro, 3',5‘-dichloro, 3‘,5‘-difluoro, 3‘,5‘-
`dimethyl and 3'-methyl-4‘-fluoro.
`Preferred examples for R3 include primary and secondary
`C1_6 alkyl and C3_6 cycloalkyl.
`Preferred examples for Y include —CH2—CH2— and
`—CH=CH—.
`
`Preferred examples for Z include
`
`HO
`
`O
`
`O
`
`O
`
`—CH(OH)CH2CH2(OH)CH2CO2R12, —CH(OH)CH2C(O)
`CHZCOZR12 and —CH(OH)CH2C(OR13)2CH2CO2R12.
`Now, more preferred substituents of the compounds of the
`present invention will be described.
`As more preferred examples for R1, R2 and R6, 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, 5-methyl, 6-methyl,
`7-methyl, 8-methyl, 5-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-butyl and
`7-dimethylamino.
`When R6 is hydrogen, R1 and R2 together represent
`6-chloro-8-methyl, 6-bromo-7-methoxy, 6-methyl-7-chloro,
`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-hydroxy,
`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-methylenedioxy, 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 R3, the above-mentioned
`preferred examples of R3 may be mentioned.
`As preferred examples for Y, —CH2—CH2— and
`—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 R6 are hydrogen, R1 is
`hydrogen, 6-methyl, 6-ethyl, 6-trifluoromethyl, 6-hydroxy,
`6-methoxy, 6-chloro, 6-bromo, 6-n-butyl 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 R3 include ethyl,
`n-propyl, i-propyl and cyclopropyl.
`Still further preferred examples for Y include
`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 R1, 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 R3 include i-propyl 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‘-yl]-hept-6-enoic acid
`(c)
`(E)-3,5-dihydroxy-7-[4'-(4“-fluorophenyl)-2‘-(1“-
`methylethyl)-6'-methyl-quinolin-3‘-yl]-hept-6-enoic acid
`(d)
`(E)-3,5-dihydroxy-7-[4‘-(4“-fluorophenyl)-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
`
`Mylan Exhibit 1001, Page 3
`
`Mylan Exhibit 1001, Page 3
`
`
`

`
`5
`
`5,856,336
`
`(e) (E)-3,5-dihydroxy-7-[4‘-(4“-fluorophenyl)-2‘-
`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
`(h) (E)-3,5-dihydroxy-7-[4‘-(4“ -fluorophenyl)-2‘-
`cyclopropyl-6',7‘-dimethoxy-quinolin-3‘-yl]-hept-6-enoic
`acid
`
`(E)-3,5-dihydroxy-7-[4‘-(4“-chlorophenyl)-2‘-(1“ -
`(i)
`methylethyl)-quinolin-3‘-yl]-hept-6-enoic acid
`(E)-3,5-dihydroxy-7-[4‘-(4“-chlorophenyl)-2‘-(1“-
`methy1ethyl)-6'-chloro-quinolin-3‘-yl]-hept-6-enoic acid
`(k) (E)-3,5-dihydroxy-7-[4'-(4“-chlorophenyl)-2‘-(1“-
`methylethyl)-6'-methyl-quinolin-3‘-yl]-hept-6-enoic acid
`(1) (E)-3,5-dihydroxy-7-[4‘-(4“-chlorophenyl)-2‘-(1“-
`methylethyl)-6',7‘-dirnethoxy-quinolin-3‘-yl]-hept-6-enoic
`acid
`
`(E)-3,5-dihydroxy-7-[4‘-(4“ -chlorophenyl)-2‘-
`(In)
`cyclopropyl-quinolin-3‘-yl]-hept-6-enoic acid
`(n)
`(E)-3,5-dihydroxy-7-[4‘-(4“-chlorophenyl)-2‘-
`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'-dirnethoxy-quinolin-3‘-yl]-hept-6-enoic
`acid
`
`(q) (E)-3,5-dihydroxy-7-[4'-phenyl-2‘-(1“-methylethyl)-
`quinolin-3‘-yl]-hept-6-enoic acid
`(r)
`(E)-3,5-dihydroxy-7-[4‘-phenyl-2‘-(1“-rnethylethyl)-
`6'-chloro-quinolin-3‘-yl]-hept-6-enoic acid
`(s)
`(E)-3,5-dihydroxy-7-[4‘-phenyl-2‘-(1“-methylethyl)-
`6'-methyl-quinolin-3‘-yl]-hept-6-enoic acid
`(t)
`(E)-3,5-dihydroxy-7-[4‘-phenyl-2‘-( 1“-rnethylethyl)-
`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)
`(E)-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‘-phcnyl-2'-cyclopropyl-6',7'-
`dimethoxy-quinolin-3‘-yl]-hept-6-enoic acid
`(y)
`(E)-3,5-dihydroxy-7-[4‘-(4“-fluorophenyl)-2‘-(1“-
`methy1ethyl)-6'-rnethoxy-quinolin-3‘-yl]-hept-6-enoic acid
`(Z) (E)-3,5-dihydroxy-7-[4‘-(4“-fluorophenyl)-2‘-
`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.
`
`R3
`
`R4
`
`R5
`R1
`
`2
`
`R
`
`COZRZI
`
`R5
`
`O
`N
`
`VII
`
`A
`
`5
`
`10
`
`15
`
`20
`
`25
`
`30
`
`35
`
`40
`
`45
`
`50
`
`55
`
`60
`
`R5
`R1
`
`2
`
`R
`
`R5
`R2
`
`1
`
`R
`
`R6
`R2
`
`1
`
`R
`
`R6
`R2
`
`1
`
`R
`
`R6
`R2
`
`1
`
`R
`
`R3
`
`-continued
`R4
`
`CHZOH
`
`O
`N
`
`R5
`
`R3
`
`VI
`
`R4
`
`CH0
`
`R5
`
`0
`N
`
`R3
`
`V
`
`R4
`
`B
`
`C
`
`/ OEt
`
`OH R
`D
`
`CHO
`
`0
`
`OH
`
`|
`
`|
`
`E
`
`COgR12
`
`F
`
`O
`N
`
`R5
`
`R3
`
`IV
`
`R4
`
`O
`N
`
`R5
`
`III
`
`R4
`
`R3
`
`O
`N
`
`11
`
`R5
`
`Mylan Exhibit 1001, Page 4
`
`Mylan Exhibit 1001, Page 4
`
`
`

`
`5,856,336
`
`R3
`
`-continued
`R4
`
`7
`-continued
`R4
`
`R3
`
`COZRIZ
`
`OH
`
`OH
`
`N
`
`R5
`
`1-1
`
`R4
`
`R3
`
`OH
`
`COZRIZ
`
`|
`
`OH
`
`N
`
`R5
`
`1-2 (R12 = H)
`1-5 (R12 = Na)
`
`R2
`
`R6
`
`1
`
`R
`
`R6
`
`R2
`
`R1
`
` 1
`
`1-3
`
`R4
`
`R3
`
`OH
`
`R5
`
`ON
`
`1-4
`
`R4
`
`R3
`
`CHO
`
`N
`
`R5
`
`R
`
`R5
`
`1
`
`R
`
`33
`
`R5
`R2
`
`10
`
`15
`
`20
`
`25
`
`30
`
`35
`
`40
`
`45
`
`50
`
`55
`
`60
`
`65
`
`R6
`R2
`
`O
`N
`
`R5
`
`R1
`
`R3
`
`VIII
`R4
`
`R6
`R2
`
`O
`N
`
`R5
`
`R1
`
`R3
`
`IX
`
`R4
`
`L
`
`M
`
`COZRZZ
`
`CHZOH
`
`CHO
`
`|
`
`|
`
`|
`
`R6
`R2
`
`1
`
`R
`
`N
`
`R5
`
`III
`
`R3
`
`R4
`
`OH
`
`R6
`
`|
`
`OH
`
`COgR12
`
`N >
`
`COgR12
`
`In the above reaction scheme, R1, R2, R3, R4, R5, RR and
`R12 are as defined above with respect to the formula I, and
`
`Mylan Exhibit 1001, Page 5
`
`Mylan Exhibit 1001, Page 5
`
`
`

`
`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-butylstannyl)
`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° C., 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)ethyl]
`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-
`
`5
`
`10
`
`15
`
`20
`
`25
`
`30
`
`35
`
`40
`
`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
`ot,[3-unsaturated carboxylic acid ester, whereby a trans-form
`ot,[3-unsaturated carboxylic acid ester can be obtained by a
`so-called Horner-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 ot,[3-
`unsaturated carboxylic acid ester to an allyl 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 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 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,[3-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 I-2 and I-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
`
`R3
`
`R4
`
`OH
`
`1-2 (R12 = H)
`1-5 (R12 = Na)
`
`R6
`R2
`
`R1
`
`R1
`
`6—OMe
`6—OMe
`6—Br
`6—Me
`7—OMe
`6—Br
`
`6,7
`
`co2R12
`
`OH
`
`|
`
`N
`
`R5
`
`R2
`
`H
`H
`H
`8—Me
`8—OMe
`H
`
`R3
`
`H
`4—F
`4—F
`4—F
`4-1:
`2-F
`
`4—F
`
`R4
`
`H
`H
`H
`H
`H
`H
`
`H
`
`R5
`
`i—Pr
`i—Pr
`i—Pr
`i—Pr
`i—Pr
`i—Pr
`
`i—Pr
`
`R5
`
`H
`H
`H
`H
`H
`H
`
`H
`
`Mylan Exhibit 1001, Page 6
`
`Mylan Exhibit 1001, Page 6
`
`
`

`
`5,856,336
`
`11
`
`TABLE 1-continued
`
`R3
`
`R4
`
`OH
`
`I-2 (R12 = H)
`I-5 (R12 = Na)
`
`COgR12
`
`OH
`
`R6
`R2
`
`R1
`
`|
`
`N
`
`R5
`
`R1
`
`H
`
`R2
`
`H
`
`R3
`
`4—F
`
`R5
`
`R4
`
`H
`
`H
`H
`6—Cl
`6-Cl
`6—OCH2Ph
`H
`H
`6-Cl
`6—Me2N
`6-Me
`6—i—Pr
`7-Me
`6—OMe
`6-Br
`6-i-Pr
`6—Cl
`5—F
`6—OMe
`6-Me
`6—Cl
`H
`H
`6—OMe
`6—OMe
`6-OMe
`6-OMe
`6-OMe
`6—Me
`6—Me
`6-Me
`6-Me
`6-Me
`6—Cl
`6—Cl
`6—Cl
`6—Cl
`6—Cl
`H
`H
`H
`H
`H
`
`H
`H
`H
`H
`H
`H
`H
`H
`H
`H
`H
`H
`H
`H
`H
`8—Cl
`6—Br
`7—OMe
`7-Me
`7—Cl
`H
`H
`7—OMe
`7—OMe
`7-OMe
`7-OMe
`7-OMe
`H
`H
`H
`H
`H
`H
`H
`H
`H
`H
`H
`H
`H
`H
`H
`
`4 —Ph
`4 —PhCH2
`4 —F
`1 -F
`4 —F
`4 —F
`1 -F
`1 -F
`4 —F
`1 -F
`4 —F
`1 -F
`4 —F
`1 -F
`1 -F
`4 —F
`4 —F
`4 —F
`1 -F
`4 —F
`4 —F
`1 -F
`H
`4 —C
`H
`4 -C
`1 -F
`H
`4 —C
`H
`1 -C
`1 -F
`H
`4 —C
`H
`4 —C
`4 —F
`H
`4 —C
`H
`4 —C
`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—Pr
`i—Pr
`c—Pr
`sec-Bu
`i—Pr
`i—Bu
`c-Pent
`c-Pent
`i—Pr
`c-Pr
`i—Pr
`c-Pr
`c—Pr
`c-Pr
`c-Pr
`c—Pr
`i—Pr
`i—Pr
`i-Pr
`i—Pr
`c—Bu
`c-Hex
`i—Pr
`i—Pr
`c-Pr
`c-Pr
`c-Pr
`i—Pr
`i—Pr
`c-Pr
`c-Pr
`c-Pr
`i—Pr
`i—Pr
`c—Pr
`c—Pr
`c—Pr
`i—Pr
`i—Pr
`c—Pr
`c—Pr
`c—Pr
`
`R5
`
`H
`
`H
`H
`H
`H
`H
`H
`H
`H
`H
`H
`H
`H
`H
`H
`H
`H
`8—Br
`8—OMe
`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
`H
`
`Further, 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.
`
`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 parcntcral 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 I. 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 for over 24 hours.
`Liver was cut out at mid-dark and microsome and superna-
`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 yl of the
`
`10
`
`15
`
`20
`
`25
`
`30
`
`35
`
`40
`
`45
`
`50
`
`55
`
`60
`
`65
`
`Mylan Exhibit 1001, Page 7
`
`Mylan Exhibit 1001, Page 7
`
`
`

`
`5,856,336
`
`14
`
`TABLE 2
`
`Inhibitory activities by Test A
`
`compound
`(Compounds of the
`present invention)
`
`Ijs,
`1-53
`(Reference compounds)
`
`Mevinolin
`C5514
`
`150 (molar concentration)
`
`1-123 X £1
`731: 10-8
`1-9 >< 10”
`
`1.4 x 10*‘
`9-0 X 107
`
`In Table 2-2, the relative activities are shown based on the
`activities of CS_514 being evaluated to be 1.
`
`TABLE 2_2
`
`Relative activities by TestA
`
`Ceiiipeiiiid
`(compounds of the
`Present iI1Venti0I1)
`H6
`H16
`1.117
`
`Reieiive eeiiviiiee
`
`1 75
`2:25
`o_37
`3%
`
`Structures of reference compounds:
`
`(1) Mevinohn
`
`0
`
`body weight (0.4 ml/100 g body weight), were dissolved in 45 «k
`
`H3C
`
`CH3
`
`0
`
`CH3
`
`
`
`Hgc“
`
`(2) cs—514
`
`NaOgC
`
`OH
`
`CH3
`
`0
`
`Hgc/ye
`
`CH3
`
`0
`
`HO
`
`Mylan Exhibit 1001, Page 8
`
`13
`reaction mixture containing ATP; 1 mM, Glutathione; 6 mM,
`Glucose-1-phosphate; 10 mM, NAD; 0.25 mM, NADP; 0.25
`mM, CoA; 0.04 mM and 0.2 mM [2-i4C]sodium acetate (0.2
`yCi) with 4 yl of test compound solution dissolved in water
`or dimethyl sulfoxide. To stop reaction and saponify, 1 ml of
`15% EtOH-KOH was added to the reactions and heated at
`
`5
`
`75° C. for 1 hour. Nonsaponifiable lipids were extracted with
`petroleum ether and incorporated i4C radioactivity was
`counted. Inhibitory activity of compounds was indicated 10
`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 15
`(FBS) at 37° C., 5% CO2 until cells were confluent for about
`7 days. Cells were exposed to the DME medium containing
`
`5% Of hpepretein deficient Serum (LPDS) Prepared by
`ultracentrifugation method for over 24 hours. Medium was
`changed to 0.5 ml of fresh 5% LpDS containing DME before 20
`assay and 10 yl of test compound solution dissolved in water
`or DMSO were added. 0.2 yCi of [2-i4C]sodium acetate (20
`yl) was added at O hr(B-1) or 4 hrs(B-2) after addition of
`compounds. After 4 hrs further incubation with [2-i4C]
`sodium acetate, medium was removed and cells were 25
`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 ml 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 30
`75° C. for 1 hour. Nonsaponifiable lipids were extracted with
`petroleum ether and i4C radioactivity was counted. Counts
`were revised by cell protein and indicated with DPM/mg
`protein. Inhibitory activity of compounds was indicated with
`IC50.
`
`35
`
`Test C: Inhibition of cholesterol biosynthesis in vivo
`
`Male Sprague-Dawley rats weighing about 150 g were fed
`normal Purina chow diet and water ad libitum, and exposed
`to 12 hours light/12 hours dark lighting pattern (2:00
`PM—2:00 AM dark) prior to use for in vivo inhibition test of
`cholesterol biosynthesis. Animals were separated groups
`consisting of five rats as to be average mean body weight in
`each groups. Test compounds at dosage of 0.02—0.2 mg/kg
`
`40
`
`water or suspended or in 0.5% methyl cellulose and orally
`administered at 2-3 hours before mid-dark (8:00 PM), while
`cholesterol biosynthesis reaches to maximum in rats. As
`control, rats were orally administered only water or vehicle.
`At 90 minutes after sample administration,
`rats were 50
`injected intraperitoneally with 10 yCi of [2-i4C]sodium
`acetate at volume of 0.2 ml per one. 2 Hours later, blood
`samples were obtained and serum were separated immedi-
`ately. Total lipids were extracted according to the method of
`Folch et al. and saponified with EtOH-KOH. Nonsaponifi- 55
`able lipids were extracted with petroleum ether and radio
`activity incorporated into nonsaponifiable lipids was
`counted.
`
`Inhibitory activity was indicated as percent decrease of
`counts in testing groups (DPM/2 ml serum/2 hours) from 60
`
`that in control group.
`
`With respect to the compounds of the present invention,
`the inhibitory activities against the cholesterol biosynthesis
`in which HMG-CoA reductase serves as a rate limiting
`enzyme, were measured by the above Test A and B. The 65
`results are shown in Tables, 2, 2-2, 3 and 3-2. Further, the
`results of the measurements by Test C are also presented.
`
`Mylan Exhi