170
`
`J . Med. Chem. 1986,29, 170-181
`
`3-Hydroxy-3-methylglutaryl-coenzyme A Reductase Inhibitors. 3.
`7 4 3,5-Disubstituted [ l,l'-biphenyl]-2-yl)-3,5-dihydroxy-6-heptenoic Acids and Their
`Lactone Derivatives
`
`G. E. Stokker,*t A. W. Alberts,$ P. S. Anderson,? E. J. Cragoe, Jr.,t A. A. Deana,t J. L. Gilfillan,t J. Hirshfield,*
`W. J. Holtz,? W. F. Hoffman,? J. W. Huff,t T. J. Lee,t F. C. Novello,t J. D. Prugh,t C. S. Rooney,' R. L. Smith,+
`and A. K. Willard+,!
`Merck Sharp & Dohme Research Laboratories, West Point, Pennsylvania 19486, and Rahway, New Jersey 07065.
`Received May 2, 1985
`The syntheses of a series of 7-(3,5-disubstituted [ l,l'-biphenyl]-2-yl)-3,5-dihydroxy-6-heptenoic acids and their lactones
`are reported. Intrinsic 3-hydroxy-3-methylglutaryl-coenzyme A reductase inhibitory activity is enhanced markedly
`when the biphenyl moiety is substituted by chloro or methyl groups at positions 3 and 5 and a fluoro group at position
`4'. These substitutions, followed by resolution, provided compounds loo(+) and 110(+) with 2.8 times the intrinsic
`inhibitory activity of compactin. Compound loo(+) was shown to possess the same chirality in the lactone ring
`as compactin by single-crystal X-ray crystallography.
`
`We previously reported on a series of 3,5-dihydroxy-
`pentanoic acids, their 6 lactones, and other derivatives
`which were shown to possess varying degrees of intrinsic
`3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) re-
`ductase inhibitory activity.' The most potent inhibitor
`bearing a monocyclic substituent was the dihydroxy acid
`form of lactone 1. In part 2, we described the structure-
`activity relationships (SAR) determined for a series of
`7-phenyl-3,5-dihydroxy-6-heptenoic (and heptanoic) acids
`bearing different aryl substituents.2 Lactone 2 emerged
`as the most interesting monosubstituted phenyl compound
`of this series and potentially the most exploitable. In the
`
`Homo
`
`-
`- -
`R
`w
`I
`
`Scheme I
`
`CHO
`
`'x
`
`Y'
`
`3
`
`4
`
`5 I'
`
`present study, we have extended the latter investigation
`by determining the effects of aryl substitution on both
`rings of a series of 7-(3,5-disubstituted [l,l'-biphenyl]-2-
`yl)-3,5-dihydroxy-6-heptenoic acids on activity. Use of the
`3,5-disubstitution pattern in the biphenyl moiety was
`based on the observations of part 2 wherein the 2,4,6-
`trisubstitution pattern on the phenyl ring was shown to
`be optimal. This report also extends the investigation of
`bridging elements between the two aryl rings with a direct
`bond, a methylene, ethylene, or ethenyl unit. The other
`bridges previously examined were oxygen, methyleneoxy,
`and oxymethylene.2
`Chemistry. The compounds prepared for this study
`are listed in Table I11 and their syntheses are summarized
`in Schemes I-IV. With the exception of 6, all of the
`
`5H
`-CH,CO-
`n-Bu,SnCH=CHOEt, n-BuLi. bH,O'.
`f H'. g C,H,CH,, A .
`-CHCO,CH,.
`NaBH,. e OH-.
`n-Bu,SnOCH,, CH,=C(OAc)CH,.
`HO,CCO,H.
`j BrCH,COBr, C,H,N.
`Zn, CuBr, Et,AlCl. H,, Rh/C.
`6-substituted 4-hydroxypyran-2-ones were synthesized
`from the appropriate aldehydes via condensation with the
`dianion of methyl acetoacetate followed by borohydride
`reduction, hydrolysis, and lactonization as described in part
`1. The characterization of the trans isomer was based on
`the chemical shift of the C-6 and the coupling constants
`(I) Stokker, G. E.; Hoffman, W. F.; Alberts, A. W.; Cragoe, E. J.,
`Jr.; Deana, A. A.; Gilfillan, J. L.; Huff, J. W.; Novello, F. C.;
`Prugh, J. D.; Smith, R. L.; Willard, A. K. J. Med. Chem. 1985,
`28, 347.
`(2) Hoffman, W. F.; Alberts, A. W.; Cragoe, E. J., Jr.; Deana, A.
`Merck Sharp & Dohme, West Point, PA.
`A.; Evans, B. E.; Gilfillan, J. L.; Gould, N. P.; Huff, J. W.;
`* Merck Sharp & Dohme, Rahway, NJ.
`Novello, F. C.; Prugh, J. D.; Rittle, K. E.; Smith, R. L.; Stokker,
`3 Present address: Stuart Pharmaceuticals, Division of IC1
`G. E.; Willard, A. K. J. Med. Chem., preceding paper in this
`issue.
`Americas, Wilmington, DE 19897.
`0022-2623/86/1829-0170$01.50/0 0 1986 American Chemical Society
`
` Mylan Exhibit 1041, Page 1
`
`

`
`HMG-CoA Reductase Inhibitors
`Scheme I1
`
`F
`
`F
`
`Journal of Medicinal Chemistry, 1986, Vol. 29, No. 2 171
`Scheme I11
`
`(Me0)zFH
`
`Br
`
`7
`
`Br
`
`8
`
`Br
`
`9
`
`10
`
`11, R = SiMez(C&)
`
`Br 4
`
`C
`
`H
`
`3
`
` t A -
`
`
`
`CH3
`12
`
`F
`
`CH3
`
`C H 3
`
`13
`
`F
`
`L
`
`A
`
`CN - A
`
`14
`15
`Br,, hu.
`ClSi(CH,),C(CH,),.
`NaOAc, A . OH-.
`Dibal.
`'-CH,CO-
`e Mg.
`f ZnBr,. g Ni(Ph,P),Cl,.
`H+. I C,H,CH,,
`f Et,B, NaBH,, -98 "C.
`-CH$O,CH,.
`(n-C,H,),NF.
`A .
`
`of the C-4 protons in the NMR as described in part 2.2
`(Since the completion of the work described in part 1, a
`trialkylborane-mediated stereoselective reduction of P-
`hydroxy ketones has been des~ribed.~ This reduction
`yields a much higher percentage of the desired erythro diol
`and an illustrative example (15a) is included in the ex-
`perimental section.) The cis-6-ethenyl-4-hydroxypyran-
`2-ones (i.e., the biologically inactive isomers) may be ep-
`imerized at c6 to a 1:1 (cis-trans) mixture by refluxing in
`aqueous acetonitrile with 1 equiv of mercuric chloride. The
`resolution of lactones 100 and 110 was accomplished via
`chromatographic separation of their respective diastereo-
`meric (S)-a-methylbenzylamides followed by basic hy-
`drolysis and relactonization to yield loo(+), loo(-), 110(+),
`and llO(-). Lactone 6 was prepared by an intramolecular
`Reformataky reaction of the antecedent 442-bromoacet-
`oxy)-6-hexen-2-one which was obtained after bromo-
`acylation of the aldol product of acetone and propenal 84
`(Scheme I).
`The synthesis of biphenylylpropenals 4 (Table 11) was
`accomplished by condensation of the corresponding bi-
`phenylcarboxaldehyde (3) with lithium ethoxyethylene
`(Scheme I) as described in part 1 with the exception of 14.
`In the latter instance, conversion of 4-bromo-2-methyl-l-
`fluorobenzene (7) to silyl ether 11 was effected via the
`
`(3) Narasaka, K.; Pai, F. C. Tetrahedron Lett. 1984, 40, 2233.
`
`I CI
`
`17
`
`16
`
`(Me 0 )2C H
`
`CHO
`
`%sph sSph
`
`18
`
`19
`
`21 CI - k , I
`
`CI
`
`20
`
`Q,
`
`2 3
`22
`a 0,. Me S. MeOH, H+. LAH. e TosC1,py.
`f C H SNa. JH,'O.
`C,H MgBr.l Me,SO, TFAA then
`E t , h 5 j NH,NH,, KOH.
`kCS. Cu2+.
`
`four-step sequence 7 - 8 - 9 - 10 - 11, using the
`
`reagents indicated in Scheme 11. Grignard formation
`followed by transmetalation with anhydrous zinc bromide
`provided arylzinc bromide A, which was immediately
`coupled with cinnamyl nitrile 12 catalyzed by bis(tri-
`pheny1phosphine)nickel di~hloride.~ Reduction of nitrile
`13 with diisobutylaluminum hydride (Dibal) provided
`aldehyde 14, the requisite biphenylylpropenal for lactone
`15. The elaboration of the only non biphenyl aldehydes
`(22 and 28) required for the dianion condensation route
`are outlined in Schemes I11 and IV, respectively.
`Ozonolysis of 16 followed by reductive workup and
`subsequent acid methanolysis provided 17, which was re-
`duced to propanol 18. conversion of 18 to the corre-
`sponding tosylate followed by displacement by thiophen-
`oxide and then acid hydrolysis yielded 19. Grignard ad-
`dition of phenylmagnesium bromide to aldehyde 19 fol-
`lowed by Swern oxidation gave benzophenone 20, which
`was then subjected to a Wolff-Kishner reduction (21) and
`a halogen-mediated oxidation to form 22.
`Scheme IV delineates the preparation of propenal 28
`from aniline 24. Meerwein arylation of methyl acrylate
`with the diazonium salt of 25 followed by treatment with
`potassium hydroxide at elevated temperature provided 26.
`Treatment of 26 with 4-chlorostyrene under Heck6 aryla-
`tion conditions yielded cinnamic acid 27, which was then
`converted to 28 via its acid chloride by reduction with
`bis(triphenylphosphine)copper(I) tetrahydroborate.6
`
`(4) Sletzinger, M.; Verhoven, T. R.; Volante, R. P.; McNamara, J.
`M.; Corley, E. G.; Liu, T. M. H. Tetrahedron Lett. 1985,2951.
`( 5 ) Plevyak, J. E.; Dickenson, J. E.; Heck, R. F. J. Org. Chem.
`1979,44, 4078.
`(6) Fleet, G. W. J.; Fuller, C. J.; Harding, P. J. C. Tetrahedron
`Lett. 1978, 1440.
`
`Mylan Exhibit 1041, Page 2
`
`

`
`172 Journal of Medicinal Chemistry, 1986, Vol. 29, No. 2
`
`Scheme IV
`"2
`
`CH3
`24
`
`Scheme V. Method 1
`C HO
`
`C02H
`
`Stokker et al.
`
`C02Me
`
`Cl
`30
`
`CI
`31
`
`CI
`32
`
`26
`
`27
`
`1
`CI
`33
`
`I
`CI
`34
`
`28
`
`H3C Au%
`
`CI
`
`29
`' Br,.
`OH-, a.
`CH,=CHCO,Et.
`HONO.
`4-C1-C,H4CH=CH , (C,H,),P, Pd(OAc),, 1
`e H,'O.
`A ((C,H,),P),CuBH,,
`mol %, Et,N, A . g SOCl,.
`(C,H,),P.
`The biphenylcarboxaldehydes 3 (Table I) were prepared
`by the two independent methods summarized in Schemes
`V and VI (method 1) and Scheme W (method 2). Method
`1, the key step of which is based on the work of Meyers,'
`involves an aryl-Grignard displacement of the 2-methoxy
`group of 36 followed by quaternization with methyl iodide,
`borohydride reduction, and acidic hydro1ysis.S No product
`resulting from displacement of the 6-chloro group was
`detected in the Grignard reaction on 36. The apparent
`inactivity of the o-chloro substituent in this displacement
`was confirmed by failure to detect any biphenylyloxazoline
`in the reaction of phenylmagnesium bromide with 2-(2,6-
`dichlorophenyl)-4,5-dihydro-4,4-dimethyloxazole under
`identical conditions. The synthesis of 35, the immediate
`precursor to 36, is outlined in Scheme V. In the original
`route (A), 30 -. 31 -. 32 -. 34 -+ 35, only the demethyl-
`ation of 32 requires comment. The nucleophilic dis-
`placement of sterically hindered carboxylates by tertiary
`amines (DBN,g D a b ~ o , ~ 3-quinnuclidin01,~ and 1,l-di-
`methylhydrazinelO) is well-known. The use of 4-(amino-
`methy1)piperidine as the nucleophile allowed a lower
`temperature (100 OC vs. 140 OC for the bicyclic amines)
`and a shorter reaction time (11/2 h vs. 6-12 h for the hy-
`drazine) to be used, and although of no real advantage in
`
`(7) Meyers, A. I.; Gabel, R.; Mihelich, E. J. Org. Chem. 1978,43.
`1372.
`(8) Nordin, I. C. J. HeterocycE. Chem. 1966, 3, 531.
`(9) Miles, D. H.; Huang, B.-S. J. Org. Chem. 1976, 41, 208 and
`references cited therein.
`(10) Kasina, S.; Nematallahi, J. Tetrahedron Lett. 1978, 1403.
`
`MeO@Ci I M e O W C l
`
`
`
`I
`I
`CI
`CI
`36
`35
`' $g,O.
`4-NH2CH,-c-C,H,,N,
`MeI, K,CO,, DMF.
`n-Bu,NI, KMnO,, C,H,, H,O. e NBS, hv.
`A .
`H,NC(CH,),CH,OH.
`SOC1,.
`
`Scheme VI. Method 1
`
`CHO
`
`CI
`3a-d
`
`' RMgX. MeI. NaBH,.
`H,O', A .
`this instance, 4-(aminomethy1)piperidine was found to be
`superior to the other amines studied for this transforma-
`tion (i.e., N-methylbenzylamine, 2,6-dimethylmorpholine,
`piperidine, 2,6-dimethylpiperidine, or 2-amino-2-methyl-
`1-propanol). Proton and 13C NMR examination of the
`amine recovered after displacement showed that only the
`secondary amine was methylated, and when 2-amino-2-
`methyl-1-propanol was used, the only methylated amine
`detected was the (NJV-dimethy1amino)propanol.
`Route B, the more expeditious of the two routes, was
`based on the key conversion of 33 -. 35 via radical bro-
`mination/oxidation and subsequent amination." The
`
`oxidation of 33 - 34 with "purple benzene", essentially
`
`(11) Cheung, Y.-F. Tetrahedron Lett. 1979, 3809
`
`Mylan Exhibit 1041, Page 3
`
`

`
`HMG-CoA Reductase Inhibitors
`
`Journal of Medicinal Chemistry, 1986, Vol. 29, No. 2 173
`
`Table I. Physical Properties of Biphenylcarboxaldehydes 3“
`A.
`
`no.
`3d
`3de
`42
`43
`44
`45h
`46
`47
`3b
`48
`3c
`
`A
`
`H
`H
`2’-Me
`3‘-Me
`4‘-Me
`3’-Et
`3’-Me0
`4’-Me0
`4‘-C1
`3’-F
`4‘-F
`
`B
`H
`H
`H
`H
`H
`H
`H
`H
`H
`H
`H
`
`X
`3-C1
`3-C1
`3-C1
`3-C1
`3-C1
`3-C1
`3-C1
`3-C1
`3-C1
`3-C1
`341
`
`Y
`541
`541
`5-Cl
`5-C1
`541
`5-C1
`5-C1
`541
`5-C1
`541
`5-C1
`
`I Y
`recryst
`solvent
`pet. ether
`n-BuC1/ hexane
`
`hexane
`
`CH2Cl2/ hexane
`hexane
`
`pet. ether
`pet. ether
`
`hexane
`hexane
`sublimed
`
`formula
`
`anal.c*d
`
`method
`mD. OC
`vield.* ’%
`48-50
`80
`1
`82-83.5
`45
`1
`70
`oil
`2
`75
`84-85
`2
`2
`61
`oil
`oil
`68
`2
`58
`97-98
`2
`64
`90-91.5
`2
`23
`89-92
`1
`2
`67
`gum
`64 (37)‘
`73-74
`1
`73 (72)’
`71-73
`2
`59
`69-71
`5-F
`3-F
`H
`4’-F
`49
`2
`5-Me
`3-Me
`H
`74
`68-70
`50
`4’-F
`2
`5-C1
`3-C1
`5’-Me
`79
`103-105
`51
`3‘-Me
`2
`5-Me
`3-Me
`5’-Me
`85
`79-81
`52
`3’-Me
`2
`5-C1
`3-C1
`4’-F
`64
`53
`2’-Me
`2
`gum
`71
`74-79
`3‘-Me
`H
`3-C1
`4’-F
`2
`54
`CH2CL2/ hexane
`5-Me
`3-C1
`4’-F
`66
`70-73
`55
`3‘-Me
`CHzC12/ hexane
`2
`541
`3-Me
`4’-F
`2
`49
`sticky solid
`56
`3‘-Me
`5-C1
`3431
`4’-F
`64
`77-79
`3’-Me
`hexane
`57
`2
`sublimed
`5-Me
`3-Me
`4’-F
`2
`82
`80-81
`3‘-Me
`58
`sub 1 i m e d
`76
`137-139
`3’,5’-Mez
`5-Me
`3-Me
`4’-F
`2
`59’
`541
`3-C1
`4’-C1
`83
`118-119
`3‘-C1
`hexane
`60
`2
`106-106.5
`hexane
`5-C1
`3-C1
`5’41
`2
`75
`3’41
`61
`5-Me
`3-Me
`4’-C1
`75
`3’-C1
`62
`hexane
`80-81
`2
`6-Me
`3-Me
`4/41
`2
`36
`3’-C1
`63
`eum
`“All of the starting benzaldehydes (30 or 39) were commercially available as were the bromides for the Grignard reaction unless so
`indicated. *Represents overall yield from Grignard reaction. e Analytical results are within f0.4% of the theoretical values unless otherwise
`noted. dlH NMR spectra were recorded on all compounds in CDC13 and the diagnostic aldehydic proton appeared as a singlet between 6 9.9
`and 10.3. Complete spectra are available as supplementary material. eEthyl bridge between aryl rings. f Anal. Calcd: C, 64.54. Found: C,
`63.76. gThis compound was isolated but not purified or analyzed before use in next step. hPope, G. W.; Bogert, M. T. J. Org. Chem. 1937,
`2, 276. ‘Represents overall yield from starting benzaldehydes 30 or 3912. ’See Experimental Section (59a).
`
`Scheme VII. Method 2 6 Y
`
`-
`
`a
`
`Y
`X
`a.2-CI 4-H
`b , 2 - F 4-F
`
`e . 2-CI
`4 - C I
`d . 2-CI 4-Me
`a . 2-Me 4 - C I
`f ,Z-Me 4-Me
`g , 2 Me 5-Me
`
`X
`
`39
`
`I 6 Y
`
`Ph
`
`
`
`
`
`X
`40
`
`mediated sequence is based on that of Murahashi.12 Four
`equivalents of triphenylphosphine per atom of palladium
`was required whenever there was a reducible halogen (Cl)
`because, with lesser amounts of the phosphine (i.e., only
`3 equiv in the preparation of 47 or 48), all of the possible
`monochloro isomers were also isolated. In addition to the
`deschloro byproducts isolated in the synthesis of 47, the
`corresponding symmetrical biphenyl (a ubiquitous by-
`product in the formation of stabilized aryl-Grignard
`reagents, a phenomenon unrelated to this particular re-
`action) was isolated along with l,l-bis(4-methoxy-
`pheny1)ethylene. The formation of the latter was inter-
`preted to result from a 2-fold addition of the Grignard
`reagent to the acetate ligand of the Pd(I1) complex fol-
`lowed by dehydration. This byproduct can be obviated
`by using the Pd(I1) complex wherein the acetate ligand is
`replaced by chloride.
`X-ray Crystallography. The absolute configuration
`of the more potent, dextrarotatory enantiomer of trans-
`lactone 100(f) was determined by X-ray crystallographic
`analysis. This enantiomer was found to have 4R,6S chi-
`rality in the lactone portion corresponding to the analogous
`centers in compactin or mevinolin; the conformation is
`illustrated in Figure 1. The dihedral angle between the
`two aryl rings (atoms Cll, Cl0, C15, and C16) is 5 4 . 7 O and
`
`(12) Murahashi, S.; Tamba, Y.; Yamamura, M.; Yoshimure, N. J.
`Org. Chem. 1978,43,4099.
`
`41
`
`3
`(3c, 42-63)
`Pd(OAc),. ABC,H,MgX, 4 equiv of
`
`a H,NC,H,.
`Ph,P.
`H,’O.
`combining routes A and B, was tried because the oxidation
`of 33 with basic Ag,O was unsuccessful.
`A more facile procedure for the preparation of 3 is shown
`in Scheme VI1 (method 2). This three-step palladium-
`
`
`
`Mylan Exhibit 1041, Page 4
`
`

`
`174 Journal of Medicinal Chemistry, 1986, Vol. 29, No. 2
`Table 11. Physical Properties of 3-Biphenylylpropenals 4
`
`C HO
`I
`
`Stokker et al.
`
`A
`
`recryst
`solvent
`hexane
`
`hexane
`
`hexane
`
`Et20
`hexane
`toluene/hexane
`hexane
`toluene/hexane
`hexane
`pet. ether
`sublimed
`
`hexane
`sublimed
`sublimed
`
`x
`formula
`anal.”Sb
`mp, “C
`yield, %
`Y
`B
`no.
`H
`64
`C15HlOC120
`C, H
`76-77
`83
`5-C1
`3-C1
`H
`gum
`66
`5-C1
`3-C1
`H
`H
`d
`65‘
`C17H14C120
`C, H
`98-99
`47
`5421
`3-C1
`H
`2’-Me
`66
`C16H12C120
`H
`67
`oil
`67
`5-C1
`3 4 1
`3’-Me
`d
`C16H12C120
`C, H
`109-111
`67
`5-C1
`3-C1
`H
`4’-Me
`68
`C16H12C120
`69
`oil
`79
`5-C1
`3 4 1
`H
`3’-Et
`d
`C17H14C120
`70
`Ci~HizC120~
`c , H
`81-82
`63
`5-C1
`3-C1
`H
`3’-Me0
`H
`71
`5421
`3-C1
`4’-Me0
`94-95
`63
`C16H12C1202
`C, H
`H
`72
`C15H9C130
`C, H
`96-97.5
`64
`5-C1
`3-C1
`4’-C1
`73
`3-C1
`H
`3’-F
`91-92
`71
`C15HgClzFO
`C, He
`5-C1
`74
`5-C1
`3-C1
`H
`4’-F
`136-138
`64
`C15HgCIZFO
`C, H
`C, H
`75
`69
`5-F
`3-F
`H
`4’-F
`81-82
`C15H9F30
`76
`C, H
`70-72
`81
`5-Me
`3-Me
`H
`4’-F
`C17H16F0
`77
`C17H14C120
`oil
`66
`5-C1
`3-C1
`Y-Me
`3’-Me
`d
`78
`Cl9H200
`gum
`74
`5-Me
`3-Me
`5’-Me
`3’-Me
`d
`C16HllC12FO
`d
`79
`gum
`61
`5-C1
`3-C1
`4‘-F
`2’-Me
`C16H12ClFO
`oil
`37
`H
`3-C1
`4’-F
`3’-Me
`80
`d
`81
`C17H14ClFO
`C, H’
`79-81
`60
`5-Me
`3-C1
`4‘-F
`3’-Me
`82
`C17H14ClFO
`oil
`40
`5-C1
`3-Me
`4’-F
`3’-Me
`d
`C16HliCI2FO
`C, H
`98.5-99.5
`84
`5-C1
`3-C1
`4‘-F
`3’-Me
`83
`84
`C, H
`82-84
`49
`5-Me
`3-Me
`4‘-F
`3’-Me
`C18H17F0
`C19H19FO
`C, H
`78-80
`70
`5-Me
`3-Me
`4’-F
`3’,5’-Me2
`85
`gum
`59
`5-C1
`3-C1
`4’-C1
`3’-C1
`86
`d
`C15H8C140
`87
`C15H8C140
`gum
`49
`5-C1
`3-C1
`5’-C1
`3’-C1
`d
`gum
`66
`5-Me
`3-Me
`4’-C1
`3’-C1
`d
`88
`C17H14C120
`89
`gum
`58
`6-Me
`3-Me
`4’-C1
`3’-C1
`d
`C17H14C120
`‘H NMR spectra were recorded on all compounds
`Analytical results are within *0.4% of the theoretical values unless otherwise noted.
`in CDC13 and the chemical shifts for the aldehydic and a-vinyl protons were 6 9.5-9.65 (d, J = 7.0-7.5 Hz) and 6.2-6.4 (dd, J = 15.C-16.0 and
`Ethyl bridge between aryl rings.
`7.0-7.5 Hz), respectively. Complete spectra are available as supplementary material.
`This compound
`was isolated but not purified or analyzed before use in next step. eAnal. Calcd: C, 61.04. Found: C, 61.47. /Anal. Calcd: H, 4.84. Found:
`H, 5.36.
`that between the central aryl ring and the ethylene bridge
`(atoms C?, C8, C9, and Clo) is 57.5’.
`Biological Results and Discussion
`The target compounds presented in Table I11 were
`tested as the ring-opened dihydroxy carboxylate forms for
`their ability to inhibit solubilized, partially purified rat
`liver HMG-CoA reductase. The study of nuclear substi-
`tution on the central phenyl ring of the biphenyl moiety
`was confined to that of methyl or chloro in the 3- and
`5-positions. Inhibitory potency increased by greater than
`40-fold when chloro groups were introduced concomitantly
`in the 3- and 5-positions (90 vs. 2). The potency of the
`3-chloro compound (106) was less than half that of the
`3,5-dichloro compound (109). Replacement of the 5-ChlOrO
`group in 109 by a methyl group (107) resulted in a slight
`reduction in potency, while transposition of these two
`
`0 21
`
`groups (107 - 108) increased the potency by 2.3-fold. In
`
`some cases, the replacement of both chloro groups by
`methyl groups resulted in a modest diminution of potency
`(100 vs. 102 and 103 vs. 104), while in others a substantial
`increase in potency was observed (109 vs. 110 and 112 vs.
`114). Replacement of the two chloro groups in compound
`100 with fluoro groups (101) resulted in a marked loss of
`potency. Movement of the methyl from position 5 (1 14)
`to position 6 (1 15) resulted in a moderate loss of potency.
`Type and position of substituents on the external phenyl
`ring were more critical. An electron-donating group in the
`4’-position (i.e., CH3 in 94 or CHBO in 97) was detrimental
`when compared to H (90), whereas a halogen in this pos-
`
`Figure 1. Computer-generated ORTEP drawing of one formula
`unit of structure loo(+) within the unit cell.
`ition was beneficial (Le., C1 in 98 or F in 100). A methyl
`in the 2’-position was also contraindicated (92 vs. 90, 105
`vs. 100, and 105 vs. 109). An electron-donating group at
`the 3‘-position may increase potency (CH3 in 93 vs. 90 and
`
`
`
`Mylan Exhibit 1041, Page 5
`
`

`
`HMG-CoA Reductase Inhibitors
`
`Journal of Medicinal Chemistry, 1986, Vol. 29, No. 2 175
`
`Table 111. Physical Properties and in Vitro HMG CoA Reductase Inhibitory Activities of Lactones 5 and 6
`1'
`
`no.
`
`A
`
`B
`
`Y'
`recryst
`solvent
`acetonelhexane
`Et20
`
`Et20/hexane
`n-BuCllhexane
`n-BuCllhexane
`n-BuCllhexane
`n-BuCllhexane
`n-BuCllhexane
`n-BuCllhexane
`
`hexane
`
`Et20/hexane
`
`n-BuC1
`
`re1
`IC50,
`X
`Y
`mp, "C
`formulaa
`potencyb
`fiM
`147-149
`C13H12C1203
`341
`5-C1
`0.06
`21.9
`1
`2
`90-92
`C1gHl&
`H
`H
`H
`H
`0.63
`3.0
`2HC
`oil
`H
`H
`H
`H
`0.04
`44
`C19H2003
`glass
`C23H25F03
`3-Me
`5-Me
`4'-F
`3'-Me
`14
`0.106
`6d
`15
`137-138
`C22H23F04
`3-Me
`n-BuC1
`5-Me
`4'-F
`3'-CH20H
`26.5
`0.049
`23'
`109-110
`CzoHzoClz03
`341
`Et20/hexane
`5-C1
`H
`H
`4.3
`0.53
`2.4
`0.63
`n-BuC1
`153-153.5 C23H23C103
`5-Me
`3-Me
`H
`4'41
`2 9
`26
`0.08
`n-BuCllhexane
`C19Hi8C1203
`5-C1
`3-C1
`H
`H
`113-115
`90
`18
`541 CHC13/hexane
`ClgHl8Cl2O3
`0.1
`151.5-152
`3-C1
`H
`H
`90"
`2.8
`C21H20C1203
`n-BuC1
`0.43
`341
`5-C1
`H
`H
`118-119
`9 18
`3.2
`CZOH18C1203
`Et20/hexane
`0.37
`H
`3-C1
`5-C1
`140-141
`2'-Me
`92
`93
`glass
`CzoH&1203*0.05CHC13
`3-C1
`541
`H
`3'-Me
`94
`0.016
`94
`119-119.5
`CzoH18C1203
`3-C1
`5-C1
`n-BuCllhexane
`H
`4'-Me
`0.19
`11
`95
`gum
`CzlH20C1203-0.05CHC13
`3-C1
`5-C1
`H
`3'-Et
`30
`0.044
`96
`93-94
`CzoH&lzO4
`3-C1
`5-C1
`H
`3'-Me0
`26
`0.05
`97
`100-102
`CzoHl&1204
`3-C1
`5-C1
`H
`4'-Me0
`2.6
`0.47
`98
`116.5-118 Ci9HI5Cl3O3
`341
`5-C1
`H
`4/41
`58
`0.036
`99
`130-132
`CigHl5Cl2FO3
`3-C1
`541
`H
`3'-F
`38.6
`0.07
`100
`153-154
`C19H15C12F03
`3-C1
`5-C1
`H
`4'-F
`100
`0.024
`loo(+)"
`108-109.5 C19Hi5C12F03
`3-C1
`541
`H
`4'-F
`280
`0.005
`loo(-)'
`130-131
`ClgHi5Cl2FO3
`3-C1
`5-C1
`H
`4'-F
`0.26
`5J
`H
`101
`glass
`3-F
`5-F
`4'-F
`3.5
`0.86
`C19H15F303
`102
`154-156
`CZ1Hz1FO3
`3-Me
`5-Me CH2C12/hexane
`H
`4'-F
`96
`0.013
`103
`glass
`C2iHzoC1203.0.05CHC13
`341
`5-C1
`5'-Me
`3'-Me
`115
`0.013
`104
`109-110
`C23H2803
`3-Me
`5-Me
`5'-Me
`3'-Me
`0.015
`80
`105
`glass
`C20H17C12F03
`3-C1
`541
`4'-F
`2'-Me
`33
`0.036
`140-145
`C20H18ClF03k
`3-C1 H
`4'-F
`3'-Me
`40
`0.03
`106
`C ~ ~ H ~ ~ C I F O :
`107
`glass
`3-C1
`5-Me
`4'-F
`3'-Me
`92
`0.013
`108
`109-110
`CzlH2oClF03
`3-Me
`541
`4'-F
`3'-Me
`233
`0.006
`109
`glass
`CzoH1,ClzF03*0.1CHC13
`3-C1
`5-C1
`4'-F
`3'-Me
`100
`0.029
`110
`115-116
`CZ2Hz3FO3
`n-BuCllhexane
`3-Me
`5-Me
`4'-F
`3'-Me
`0.007
`171
`llO(+)m 3'-Me
`87-89
`C22H23F03
`3-Me
`5-Me Et20/hexane
`4'-F
`289
`0.0076
`110(-)"
`86-88
`C22H23F03
`3-Me
`5-Me Et20/hexane
`4'-F
`3'-Me
`0.18J
`7.3
`llOHc
`131.5-133
`Cz&$"3°
`3-Me
`5-Me
`n-BuC1
`4'-F
`3'-Me
`45
`0.022
`111
`142-146
`CZ3Hz5FO3
`3-Me
`5-Me CH2C12/hexane
`4'-F
`3',5'-Me2
`150
`0.01
`112
`glass
`3-C1
`5-C1
`4'-C1
`3'-C1
`116
`0.025
`C19H14C1403
`113
`glass
`C19H14C1403
`3-C1
`541
`5'-C1
`3'-C1
`78
`0.037
`128-129
`C21HzoC1203
`3-Me
`5-Me
`4'-C1
`3'-C1
`175
`0.008
`114
`123
`0.013
`115
`glass
`3-Me
`6-Me
`4'-C1
`3'-C1
`C21H20C1203
`"Analytical results are within f0.470 of the theoretical values unless otherwise noted. bPotency of compactin arbitrarily assigned a value
`of 100; see part I for full description of protocol. cSaturated bridge between biphenyl and lactone moieties. d4-Methyl in lactone ring.
`eMethylene bridge between aryl rings (from 22). 'Ethylene bridge between aryl rings (from 28). #Ethyl bridge between aryl rings (from 3d).
`[a]20; +38.80 (CHC13). ' [aIz0D -39.85 (CHCl,). 'Each enantiomer was found to be free of the other enantiomer within the limits of
`detection (threshold = ca. 2%). Therefore, the activity displayed by the (-) enantiomer was probably due to trace amounts of the (+)
`enantiomer. kExact mass, calcd m / e 360.81; found m / e 360.089. lExact mass, calcd m / e 374.83; found m / e 374.1093. " [ a I z 5 D +39.3
`(CHC13); +36.0 (CH,OH). " CY]^^^ -35.8 (CH30H). "Anal.
`Calcd: C, 74.13. Found: C, 73.56.
`110 vs. 102) or have no effect (CH, in 109 vs. 100 or CH,O
`be made because the corresponding biphenyl was not
`prepared, the likelihood that the latter would be less potent
`in 96 vs. 90 and C2H5 in 95 vs. 90). The later case dem-
`than compound 29 can be inferred. The replacement of
`onstrated that homologation of the 3'-methyl group at-
`tenuated potency (95 vs. 93) as did oxidation to the alcohol
`chloro groups in the central phenyl ring by methyls pro-
`duced a change in potency by a factor between 0.7 and 1.7
`(15 vs. 110). The addition of a halogen at the 3'-position
`(vida supra). Therefore, the 20-fold decrease in potency
`potentiated potency but not as effectively as when intro-
`between 29 and 98 must, in a large part, be due to the
`duced at the 4'-position (99 vs. 100 or 112 vs. 98 vs. 90 and
`ethenyl bridge.
`113 vs. 112).
`Without exception, saturation of the ethenyl bridge
`The type of bridging between the phenyl rings also
`between the lactone and biphenyl moieties was detrimental
`proved to be important. When the direct bond in com-
`(2H, 90H, and llOH), a result that is opposite to similar
`pound 90 was replaced by oxygen, methyleneoxy, or oxy-
`cases in the 6-benzyl ether series.2
`methylene, the potency was decreased by 1% 7- and Cfold,
`The addition of a methyl group to the 4-position of 110
`respectively.2 Insertion of a methylene unit between the
`to give 6, a compound that more closely resembles the
`phenyl rings caused a 4-fold reduction in potency (23 vs.
`HMG moiety of the substrate HMG-CoA, lowered potency
`90H), while insertion of an ethylene unit caused a 10-fold
`12-fold. An identical addition to 1 (a much less potent
`reduction (91 vs. 90). Although no direct comparison
`inhibitor) had little effect.' The contribution of the lactone
`between an ethenyl bridge (29) and a direct bond could
`
`
`
`Mylan Exhibit 1041, Page 6
`
`

`
`176 Journal of Medicinal Chemistry, 1986, Vol. 29, No. 2
`Stokker et al.
`6- (4’-Fluoro-3,3’,5-trimet hyl[ 1,l’- biphenyl]-2-yl)-2-oxo-5-
`moiety stereochemistry to intrinsic inhibitory activity was
`hexen-4-yl2-Bromoacetate (6b). 2-Bromoacetyl bromide (83
`shown earlier to be very important in that all of the activity
`YL, 1.0 mmol) was added to a stirred solution of 6a (300 mg, 0.92
`resides in one of the enantiomers of the trans
`mmol) and pyridine (81 pL, 1.0 mmol) in Et20 (20 mL) at 0 “C.
`This observation was extended and confirmed in this study
`The ice bath was removed and the reaction mixture was stirred
`by resolving lactones 100 and 110 to afford enantiomers
`at 20 “C for 2 h and then diluted with H 2 0 (100 mL) and ad-
`loo(+) and 110(+), each of which had about 2.8 times the
`ditional Et20 (100 mL). The organic layer was separated and
`intrinsic inhibitory potency of compactin.
`washed with 1 N HCl(50 mL), H20 (2 X 100 mL), and saturated
`Two independent chiral syntheses of 110(+) have re-
`brine, dried, filtered, and evaporated. The residual oil was
`cently been published4J3 and the in vivo activity will be
`chromatographed on silica gel. Elution with CH2C1,-acetone (991,
`v/v) provided 6b (270 mg, 66%) as a viscous, pale yellow oil; NMR
`described elsewhere.
`6 2.1 (3 H, s), 2.3 (9 H, br s), 2.35-2.75 (2 H, m), 3.7 (2 H, s), 5.36
`Conclusion
`(H, dd, J = 16.5 and 6 Hz), 5.55-5.8 (H, m), 6.6 (H, d, J = 16.5
`Analysis of the intrinsic inhibitory potencies of the
`Hz), 6.9-7.2 (5 H, m).
`Compound 6. A solution of 6b (250 mg, 0.56 mmol) in dry
`compounds evaluated in this study suggests that inhibitory
`THF (10 mL) was added dropwise to a vigorously stirred slurry
`binding to HMG-CoA reductase is augmented by (a) a 3-
`of activated Zn dust (60 mg, 0.85 mmol), CuBr (10 mg, 0.07 mmol),
`and 5-chloro or -methyl group on the central phenyl ring
`Et2AlCl (25% solution in hexane; 0.34 mL, 0.6 mmol), and dry
`and (b) a 3’-methyl and 4‘-halo group on the external
`THF (5 mL) under N, at 20 “C. Stirring was continued for 5 h
`phenyl ring, while binding is decreased by (c) increasing
`before quenching with pyridine (1 mL) followed by addition of
`the distance between the two phenyl rings, (d) saturation
`H2O (100 mL) and Et20 extraction (3 X 50 mL). The combined
`of the ethenyl bridge between the biphenyl and lactone
`Et20 extracts were washed with 1 N HC1 (3 x 50 mL), H20 (2
`moieties, and (e) introduction of a methyl at the 4-position
`x 100 mL) and saturated brine, then dried, filtered, and evapo-
`of the lactone ring. The absolute stereochemistry of the
`rated, leaving a golden glass (200 mg) which was a mixture of the
`cis and trans isomers of 6. The isolation of the pure trans isomer
`lactone ring must be the same as in compactin and mev-
`was accomplished by HPLC on a Whatman M9/50 Partisil PAC
`inolin; in the present case, 4R,6S.
`column (10 X 20 mg injections). Elution with i-PrOH-hexane
`(1:20, v/v) at 8 mL/min provided 6 (70 mg, 34%) as a clear
`Experimental Section
`colorless glass. Elution times on this column under these con-
`Melting points were determined on a Thomas-Hoover capillary
`ditions were 20 min for the trans isomer and 26.4 min for the cis
`melting point apparatus and are uncorrected. Solutions were dried
`isomer; NMR 6 1.35 (3 H, s), 1.4-2.0 (2 H, m), 2.28 (3 H, s), 2.35
`over anhydrous MgSO, and evaporated under reduced pressure
`(6 H, s), 2.4-2.8 (2 H, m), 5.0-5.4 (H, m), 5.4 (H, dd, J = 16.5 and
`(rotary evaporator). ‘H NMR spectra were recorded in CDCl,
`6 Hz), 6.6 (H, d, J = 16.5 Hz), 6.9-7.2 (5 H, m).
`(unless otherwise noted) on either a Varian T-60, EM-390, or
`4-Bromo-2-(bromomethyl)-l-fluorobenzene (8). To a re-
`NT-360 spectrometer. Chemical shifts are reported in parts per
`fluxing solution of 5-bromo-2-fluorotoluene (3.8 g, 20 mmol) in
`million relative to Me4Si as the internal standard. Elemental
`CC4 (30 mL) illuminated with a 275-W UV-sunlamp, a solution
`analysis for carbon, hydrogen, and nitrogen were determined with
`of Br2 (1.1 mL, 20 mmol) in CCl, (30 mL) was added dropwise.
`a Perkin-Elmer Model 240 elemental analyzer and are within
`Refluxing and irradiation were continued for an additional 0.5
`*0.4% of theory unless noted otherwise. Optical rotations were
`h, and then the clear pale amber solution was concentrated.
`determined with a Perkin-Elmer Model 141 polarimeter. All
`Distillation of the residue provided 8 as a clear colorless oil (4.3
`starting materials were commercially available and used as re-
`g, 80%); bp 126-136 “C (15 mm); NMR 6 4.38 (2 H, s), 6.6-7.7
`ceived unless so indicated.
`3-([ l,l’-Biphenyl]-2-yl)-2-propenals (4). These compounds
`(3 H, m).
`24 Acetoxymet hy1)-4-bromo- 1 -fluoroben zene (9). Anhyd-
`(64-89) were prepared by the general method described in ref 1;
`rous sodium acetate (1.65 g, 20 mmol) was added to a solution
`their physical properties are listed in Table 11.
`trans -6-[2-( [ l,l’-Biphenyl]-2-yl)ethenyl]-3,4,5,6-tetra-
`of 8 (3.9 g, 14.6 mmol) in DMF (25 mL) and the reaction mixture
`hydro-4-hydroxy-2H-pyran-2-ones (5). These compounds (15,
`was stirred under N2 on a steam bath for 11 h. The reaction
`23,29,90-115) were elaborated from the corresponding propenals
`mixture was cooled and distributed between H 2 0 (200 mL) and
`E t 0 (100 mL). The organic layer was separated and washed with
`by the general method described in ref 1; their physical/biological
`H 2 0 (3 X 100 mL), dried, filtered, and evaporated, leaving 9 as
`properties are listed in Table 111.
`Hydrogenation of Lactones (5). Lactones 5 were hydro-
`a nearly colorless oil (3.2 g, 100%); NMR 6 2.08 (3 H, s), 5.08 (2
`H, s), 6.7-7.9 (3 H, m).
`genated over Rh/C in an analogous manner to the procedure
`5-Bromo-2-fluorobenzyl Alcohol (10). A solution of 9 (3.2
`described in ref 1; their physical/biological properties are listed
`in Table I11 (2H, 90H, and llOH).
`g, 14.5 mmol), EtOH (20 mL), and 1 N NaOH (20 mL, 20 mmol)
`(E)-6-[2-(4’-Fluoro-3,3’,5-trimethyl[ l,l’-biphenyl]-2-y1)-
`was stirred at reflux under N2 for 1.5 h. The reaction mixture
`et henyl]-3,4,5,6-tetrahydro-4-hydroxy-4-methyl-2H-pyran-
`was cooled and distributed between H20 (150 mL) and Et20 (150
`2-one (6). (a) 6-(3,3’,5-Trimethyl-l’-fluoro[ 1,l’-biphenyl]-2-
`mL). The organic layer was separated and washed with H20 (2
`y1)-4-hydroxy-5-hexen-%-one (6a). 2-Acetoxypropene (1.8 g, 18
`x 100 mL) and dried and the clear faint yellow solution was
`evaporated. Distillation of the residue provided 10 as a clear
`mmol) and tributyltin methoxide (4.8 g, 15 mmol) were combined
`colorless oil (1.7 g, 57%); bp 82-87 “C (0.4 mm); NMR 6 3.3 (H,
`and stirred at 60-70 “C under N2 for 1 h and then placed under
`vacuum for an additional 30 min. Propenal 75 (3.2 g, 12 mmol)
`br s), 4.6 (2 H, s), 6.81 (H3, t, J = 9 Hz), 7.15-7.37 (H4, m), 7.44
`(Hs, dd, J = 3 and 6 Hz).
`was added and the reaction mixture was stirred on a steam bath
`4-Bromo-l-fluoro-2-[ [ [ (1,l-dimethylethyl)dimethylsilyl]-
`under N2 for 4 h. The clear reaction mixture was then cooled,
`oxy]methyl