United States Patent
`
`[19]
`
`[11] Patent Number:
`
`6,066,650
`
`Biller et al.
`
`[45] Date of Patent:
`
`May 23, 2000
`
`USOU6U'6665UA
`
`
`
`........................ .. 5147309
`571993 Pegiion e1 :11.
`5.203.243
`5147314
`571993 Musscr el al.
`5,212,132
`5147252
`671993 Baldwin et a1.
`5,215,989
`5147253
`.
`1171993 Van1_)ae1ecta1.
`5.262.418
`_ 5147255
`571995 reiasuda e1a1_ _______ __
`5,52'7,811l
`11719911 De Cosla et al.
`.................... .. 5147408
`5,571,332
`1171996 Gluchowskicl al.
`5147318
`5,578,611
`‘
`,
`‘
`,
`_
`_
`‘
`I‘0R'¢1UN P4114“ 1' 130CUM13N1S
`05844467-‘K2
`371994 European Pal. Off. .
`D643n5—,.Al
`“I995
`Fumpean Pal Ofi.
`woe ' 40 '40
`-
`1”
`1’
`121199“ WW0‘
`OTHER PUBLICATIONS
`_
`_
`,
`_
`,
`_
`Archibald oi al., Bonzamidopiporidines. 3. Carbocyclic
`I)c1'iva1ivt:s Relaled 10 Indoramin,
`.10llI‘1'1211 of Medicinal
`
`Chemislry, vol. 17, No. 7, pp. 739-744, .1111. 1974.
`
`(List continued on next page.)
`
`Prirriary Exar11i11er—Riehard L. Raymond
`Assistant 1':.'xar11iner—Brenda Coleman
`
`Aflr0_,-fie); Agemr} or Ft}-m_Bun0n Rodney
`
`I 57'
`
`-
`
`ABS'[‘RAC]‘
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`‘
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`rgrlfsoaunncairiitgglEghifiiiiigosls and related dlSCaScS' The
`‘
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`
`[54]
`
`INHIBITORS 01" MICROSOMAL
`TRIGLYCERIDE TRANSFER PROTEIN AND
`ME'l1H0D
`
`[75]
`
`Inventors: Scott A. Biller, Hopewell; John K.
`Dickfion. Easlampmn. bolh of N.J-; 11-
`Michael L€IW1“el1€9,Yaf<11¢y, Pa; David
`R. Magnin, Ilamillon, N..I.; Michael/\.
`Poss, I.aw1'enceville, N.J.; Richard B.
`S';'l5kys‘Fr‘1nklin Parka N-:1-§ Joseph A‘
`Tlno, I_.1wrenccv11le, N..I., John E.
`»
`-
`-
`‘v
`Richard A. Partyka, Nes.hani1.',N..I.
`_
`_
`_
`[73] Asslgnflfll Bristol-Myers Squibb Company,
`Princeton, Nu],
`
`[21]
`
`/\pp1_ N;-,_; l]8,7898,3l]3
`
`1221
`
`Filed:
`
`Jll1- 21, 1997
`
`Related U-S- APPHC390“ D313
`
`[63] Continualion of applicalion No. 0874727167, Jun. 6, 1995,
`Pat. No. 5,739,135, which is a eo11tinuatio11—i11—parl of appli-
`cation No. 087391,901, Feb. 21, 1995, abandoned, which is
`21 ¢:0nlinuz1tion—in—par‘L of applicalion No. 08,084,808, Aug.
`5,1994,-bd
`d_.h"h'.-
`-1‘v1‘—‘—v1r
`872.
`
`[51]
`
`Int. Cl.7 ........................ A6'1K 317445; A6lK 31747;
`A61K 31752; A61K 317495; C07D 211700;
`C07D 217700; C07D 215738; 007D 401700
`........................ .. 5147315; 5147248; 5147252;
`[52] U.S. Cl.
`5147266; 5147307; 5147314; 5147318; 5147320;
`5147321; 5147322; 5147323; 5147324; 5147326;
`5147329; 5447235; 5447238; 5447277; 5447407;
`5467146; 5467169; 5467194; 5467196; 5467197;
`5467198; 5467199; 5467201; 5467202; 5467208;
`5467209; 5467211; 5467213; 5467214; 5467223;
`5467224
`
`[58] Field of Search ................................... .. 5147248, 252,
`5147266, 307, 314, 315, 318, 320, 321,
`323, 322, 324, 326, 329; 5447235, 238,
`277, 407; 5467146, 169, 194, 196, 197,
`198, 199, 201, 202, 208, 209, 211, 213,
`214, 224, 223
`
`[56]
`
`References Cited
`
`U.S. P/\lLN'l' 1)OCUM1_".N'l'S
`
`................... .. 260729362
`1071975 Cavalla et a1.
`971981 Benglsson el al.
`4247267
`171983 Boix—Ig1casias cl al.
`4247267
`171986 Archiliald et al.
`5147319
`371986 Tallara cl a1.
`5147212
`471986 Tallara cl a1.
`5147212
`871986 Pierce ......... ..
`5147323
`571989 Pieciola et al.
`5447391
`471990 Ruger et al.
`........... ..
`5147255
`671991 Vegz1—Nover0|a etal.
`5467224
`771991 Desai et :11.
`............ ..
`5147321
`
`.
`
`
`
`741991 Baldwin ‘*1 “L “
`371992 Desai et :11.
`771992 Pan cl a1.
`271993 Peglion c1 :11.
`
`5147313
`5147324
`5147460
`........................ .. 5147319
`
`wherein R1 lo R7, Q, X and Y are as defined herein.
`
`2 Claims, N0 Drawings
`
`1 of 162
`
`PENN EX. 2216
`
`CFAD V. UPENN
`
`IPR2015-01836
`
`3,910,931
`4,289,781
`4,367,232
`4,563,466
`4,576,940
`4,581,355
`4,607,042
`4,826,975
`4,918,073
`5,026,858
`5,028,616
`
`5vU32=593
`5,098,915
`5,130,333
`5,189,045
`
`

`
`6,066,650
`Page 2
`
`()'l'1Il5-"R PUBLIC/\'l'I()NS
`
`Bulleid & Freedman, Nature 335, 649-651 (1988). “Defec-
`tive co-translational formation of disulphide bonds in pro-
`tein disulphideisomerase-deficient microsomes”.
`Koivu et al., J. Biol. Chem. 262, 6447-6449 (1987). “A
`Single Polypeptide Acts Both as the
`Subunit of Prolyl
`4-Ilydroxylase and as a Protein Disullide-Ison'1erase*”.
`Kane & Havel in the Metabolic Basis of Inherited Disease,
`Sixth Edition, 1139-1164 (1989). “Disorders of the l3iogen-
`esis and Secretion of Lipoproteins Containing The B Apo-
`lipoproteins”.
`Schaefer et al., Clin. Chem. 34, B9-B 12 (1988). “Genetics
`and Abnormalties in Metabolism of Lipoproteins”.
`Drayna et al., Nature 327, 632-634 (1987). “Cloning and
`sequencing of human cholesteryl ester
`transfer protein
`c[)NA”.
`
`Pihlajaniemi et a1., BMBO J. 6, 643-649 (1987). “Molecular
`cloning of the [3-subunit of human prolyl 4-hydroxylase.
`This subunit and protein disulphide isomerase are products
`of the same gene".
`Yamaguchi et al., Biochem. Biophys. Res. Comm. 146,
`1483-1492 (1987). “Sequence of Membrane-Associated
`Thyroid Hormone Binding Protein From Bovine Liver: Its
`Identity with Protein Disulphide lsomerase”.
`Edman et al., Nature 317, 267-270 (1985). Sequence of
`protein disulphide isomerase and implications of its rela-
`tionship to thioredoxin.
`Kao et al., Connective Tissue Research 18, 157-174 (1988).
`“Isolation of cDNA Clones and Genomic DNA Clones of
`
`[5-Subunit of Chicken Prolyl 4-Hydroxylase*”.
`Wetterau, J. et al., Biochem 30, 9728-9735 (1991). “Protein
`Disulfide Isomerase Appears Necessary To Maintain the
`Catalytically Active Structure of the Microsomal Triglycer-
`ide Transier Protein".
`
`Morton, R.E. et al., J. Biol. Chem. 256, 1992-1995 (1981).
`“A Plasma Inhibitor of Triglyceride and Chloesteryl Ester
`Transfer Activities”.
`
`Wetterau, J. et alz, lliochem: 30, 4406-4412 (1991): “Struc-
`tu ral Properties of the Microsomal Triglyceride-Transfer
`Protein Complex”.
`Wetterau, J. et al., J. Biol. Chem. 265, 9800-9807 (1990).
`“Protein Disulfide Isomerase Is
`a Component of
`the
`Microsomal Triglyceride Transfer Protein Complex”.
`Wetterau, .l. and Zilversmit, D.B., Chem. and Phys. of Lipids
`38, 205-222 (1985). “Purification and Characterization of
`Microsomal Triglyceride and Cholesteryl Ester Transfer
`Protein From Bovine Liver Microsomes”.
`
`Wetterau, C. and Zilversmit, D.B., Biochimica et Biophysica
`Acta 875, 610-617 (1986). “Localization of intracellular
`triaeylglycerol and cholesteryl ester transfer activity in rat
`tissues”.
`
`Wetterau, J. and Zilversmit, D.B., J. Biol. Chem. 259,
`10863-10866 (1984). “A Triglyceride and Cholesteryl Ester
`Transfer Protein Associated with Liver Microsomes”.
`
`Wetterau,.J., Grant Application entitled: "Intracellular Trig-
`lyceride Transport and Metabolism”, 1987.
`Presentation Materials, Aspen Bile Acid;’Cholesterol Con-
`ference, Aug. 15, 1992.
`Wetterau, J. R., et al., Science, vol. 258, 999-1001, Nov. 6,
`1992 “Absence of Microsomal Triglyceride Transfer Protein
`in Individuals with Ahetalipoproteinemia".
`Archibald, J. L., et al., Journal of Medicinal Chemistry, vol.
`14, No. 11, pp. 1054-1059, 1971.
`
`Cortizo, L. et al., J. Med. Chem., 34, pp. 2242-2247, 1991.
`Hall, I. H. et al., Pharmaceutical Research, vol. 9, No.10, pp.
`1324-1329 1992.
`
`Ilall, I. II., et a1., Phannacological Research Communica-
`tions, vol. 19, No. 12, pp. 839-858, 1987.
`Murthy et al., Eur. J. Med. Chem .—Chim. Ther., vol. 20, No.
`6, pp. 547-550, 1935.
`Denvent Abstract No. 93-117225114, 1993.
`
`2 of 162
`
`PENN EX. 2216
`CFAD V. UPENN
`
`lPR2015-01836
`
`

`
`6,066,650
`
`1
`INHIBITORS OF MICROSOMAI.
`TRIGLYCERIDE TRANSFER PROTEIN AND
`METHOD
`
`CR[)SS-Rl:‘.lil_7Rl_-'N(Il:‘. T() REI./\'l'El)
`Al-’PI.ICATIONS
`
`This is a continuation of application Ser. No. 08;’472,067,
`filed Jun. 6, 1995 now U.S. Pat. No. 5,739,135, which is a
`continuation—in—part of application Ser. No. 08X391,901,
`filed Feb. 21, 1995, now abandoned, which is
`a
`continuation—in—part of application Ser. No. 08;’284,808,
`filed Aug. 5, 1994, now abandoned, which is a continuation-
`in—part of application Ser. No. 08;’11'.r',362, filed Sep. 3,
`1993, now U.S. Pat. No. 5,595,872.
`
`10
`
`l-‘IL-'.I.[) Oi‘ 'l‘lIl:‘. INVENTION
`
`This invention relates to novel compounds which inhibit
`microsomal triglyceride transfer protein, and to methods for
`decreasing serum lipids and treating atherosclerosis employ-
`ing such compounds.
`
`20
`
`B/\CKGR()UNlJ ()1-‘ TIIE INVl:'.N'l'lON
`
`The microsomal triglyceride transfer protein (MTP) cata-
`lyzes the transport of triglyceride (TG), cholesteryl ester
`(C13,), and phosphatidylcholine (PC) between small unila-
`mellar vesicles (SUV). Wetterau & Zilversmit, Chem. Pirys.
`Lipids 38, 205-22. (1985). When transfer rates are expressed
`as the percent of the donor lipid transferred per time, MTP
`expresses a distinct preference for neutral lipid transport
`(PG and (IE), relative to phospholipid transport. The protein
`from bovine liver has been isolated and characterived.
`
`I.r'_p£ds 38, 205-22
`Wetterau & Zilversmit, Chem. Phys.
`(1985). Polyacrylamide gel electrophoresis (PAGE) analysis
`of the purified protein suggests that the transfer protein is a
`complex of two subunits of apparent molecular weights
`58,000 and 88,000, since a single band was present when
`purified MTP was electrophoresed under nondenaturing
`condition, while two bands of apparent molecular weights
`58,000 and 88,000 were identified when electrophoresis was
`performed in the presence of sodium dodecyl sulfate (SDS).
`These two polypeptides are hereinafter referred to as 58 kDa
`and 88 kDa, respectively, or the 58 kDa and the 88 kDa
`component of MTP, respectively, or the low molecular
`weight subunit and the high molecular weight subunit of
`MTP, respectively.
`Characterization of the 58,000 molecular weight compo-
`nent of bovine MTP indicates that
`it
`is the previously
`characterized multifunctional protein, protein disulfide
`isomerase (PDI). Wetterau et al.,J. Biol. Chem. 265, 9800-7
`(1990). The presence of PDI
`in the transfer protein is
`supported by evidence showing that (1) the amino terminal
`25 amino acids of the bovine 58,000 kDa component of
`MTP is identical to that of bovine PDI, and (2) disulfidc
`isomerase activity was expressed by bovine MTP following
`the dissociation of the 58 kDa—88 kDa protein complex. In
`addition, antibodies raised against bovine PDI, a protein
`which by itself has no TG transfer activity, were able to
`immunoprecipitate bovine TG transfer activity from a solu-
`tion ccntaining purified bovine MTP.
`PDI normally plays a role in the folding and assembly of
`newly synthesized disulfidc bonded proteins within the
`lumen of the endoplasmic reticulum. Bulleid & Freedman,
`Norm-e 353, 649-51 (1988). It catalyzes the proper pairing
`of cysteine residues into disulfidc bonds, thus catalyzing the
`proper folding of disulfidc bonded proteins. In addition, PDI
`
`30
`
`40
`
`50
`
`55
`
`60
`
`65
`
`2
`has been reported to be identical to the beta subunit of
`human prolyl 4—hydroxylase. Koivu et al., J. Biol. Chem.
`262, 6447-9 (1987). The role of l-‘DI in the bovine transfer
`protein is not clear.
`It does appear to be an essential
`component of the transfer protein as dissociation of PDI
`from the 88 kDa component of bovine MTP by either low
`concentrations of a denaturant (guanidine HCI), a chaotropic
`agent (sodium perchlorate), or a ncndenaturing detergent
`(octyl glucoside) results in a loss of transfer activity. Wet-
`terau et al., Biochevmlsfry 30, 9728-35 (1991).
`Isolated
`bovine PDI has no apparent lipid transfer activity, suggest-
`ing that either the 88 kDa polypeptide is the transfer protein
`or that it confers transfer activity to the protein complex.
`The tissue and subcellular distribution of MTP activity in
`rats has been investigated. Wetterau & Zilversmit, Biochem.
`Biophys./tern 875, 610-7 (1986). Lipid transfer activity was
`found in liver and intestine. Little or no transfer activity was
`found in plasma, brain, heart, or kidney. Within the liver,
`MTP was a soluble protein located within the lumen of the
`microsomal fraction. Approximately equal concentrations
`were found in the smooth and rough microsomes.
`Abetalipoproteinemia is an autosomal recessive disease
`characterized by a virtual absence of plasma lipoproteins
`which contain apolipoprotein B (apoB). Kane & Ilavel in
`Tire Metrrbofic Hrrsis of Inherited Di.s'eo.s'e, Sixth edition,
`1139-64 (1989). Plasma TC} levels may be as low as a few
`mg/dl., and they fail
`to rise after fat
`ingestion. Plasma
`cholesterol levels are often only 20-45 mg.»’dI.. These abnor-
`malities are the result of a genetic defect in the assembly
`andfor secretion ofvery low density lipoproteins (VI.[)I.) in
`the liver and chylomicrons in the intestine. The molecular
`basis for this defect has not been previously determined. In
`subjects examined, triglyceride, phospholipid, and choles-
`terol synthesis appear normal. At autopsy, subjects are free
`of atherosclerosis. Schaefer et al., Ciin. Chem. 34, 139-12
`(1988). A link between the apoB gene and abetalipopro—
`teinemia has been excluded in several families. Talmud et
`
`al.,J'. (.'fin. i'nve.s't. 82, 1803-6 (1988) and Iluang et al.,Am.
`J. Hrmi. Genet. 46, 1141-8 (1990).
`Subjects with abetalipoproteinemia are alllicted with
`numerous maladies. Kane & Havel, supra. Subjects have fat
`malabsorption and TG accumulation in their enterocytes and
`hepatocytes. Due to the absence of TC}-rich plasma
`lipoproteins, there is a defect in the transport of fat-soluble
`vitamins such as vitamin 1:1. This results in acanthocytosis of
`erythrocytes, spinocerebellar ataxia with degeneration of the
`fasciculus cuneatus and gracilis, peripheral neuropathy,
`degenerative pigmentary retinopathy, and ceroid myopathy.
`Treatment of abetalipoproteinernir: subjects includes dietary
`restriction of fat intake and dietary supplementation with
`vitamins A, E and K.
`In vitro, MTP catalyzes the transport of lipid molecules
`between phospholipid membranes. Presumably,
`it plays a
`similar role in vivo, and thus plays some role in lipid
`metabolism. The subcellular (lumen of the microsomal
`fraction) and tissue distribution (liver and intestine) of MTP
`have led to speculation that it plays a role in the assembly of
`plasma lipoproteins, as these are the sites of plasma lipo-
`protein assembly. Wetterau & Zilversmit,BrToehem. Biophys.
`/tcm 875, 610-7 (1986). The ability of MTP to catalyze the
`transport of TG between membranes is consistent with this
`hypothesis, and suggests that MTP may catalyze the trans-
`port of TG from its site of synthesis in the endoplasmic
`reticulum (ER) membrane to nascent lipoprotein particles
`within the lumen of the ER.
`
`Olofsson and colleagues have studied lipoprotein assem-
`bly in HepG2 cells. Bostrom et al., J. Biol. Chem. 263,
`
`3 of 162
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`
`3
`4434-42 (1988). Their results suggest small precursor lipo-
`proteins become larger with time. This would be consistent
`with the addition or transfer of lipid molecules to nascent
`Iipoproteins as they are assembled. MTP may play a role in
`this process.
`In support of this hypothesis, Howell and
`Palade, J. Cell Him’. 92, 833-45 (1982), isolated nascent
`lipoproteins from the hepatic Golgi fraction of rat
`liver.
`There was a spectrum of sizes of particles present with
`varying lipid and protein compositions. Particles of high
`density lipoprotein (HDL) density, yet containing apoB,
`were found. Higgins and Hutson, J.
`lipid Res. 25,
`1295-1305 U984),
`reported lipoproteins isolated from
`Golgi were consistently larger than those from the endo-
`plasmic reticulum, again suggesting the assembly of lipo-
`proteins is a progressive event.
`
`Recent reports (Science, Vol. 258, page 999, 1992; I).
`Sharp et. al., Nature, Vol. 365, page 65, 1993) demonstrate
`that the defect causing abetalipoproteinemia is in the MTP
`gene, and as a result, the MTP protein. Individuals with
`abetalipoproteinernia have no MTP activity, as a result of
`mutations in the MTP gene, some of which have been
`characterized. These results indicate that MTP is required for
`the synthesis of apoB containing lipoproteins, such as
`VI ,DI.,
`the precursor to I.[)I..
`It
`therefore follows that
`inhibitors of MTP would inhibit the synthesis of VI.|)I . and
`LDL, thereby lowering VLDL levels, LDL levels, choles-
`terol levels, and triglyceride levels in animals and man.
`
`Canadian Patent Application No. 2,091,102 published
`Mar. 2, I994 (corresponding to US. application Ser. No.
`117,362,
`filed Sep. 3, 1993 (file DC21b)) reports MTP
`inhibitors which also block the production of apoB contain-
`ing lipoproteins in a human hepatic cell line (HepG2 cells).
`This provides further support for the proposal that an MTP
`inhibitor would lower apoB containing lipoprotein and lipid
`levels in vivo. This Canadian patent application discloses a
`method for identifying the MTP inhibitors
`
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`6,066,650
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`SUMMARY ()1-'
`
`4
`'I'IIl:‘. INVILNTION
`
`invention, novel com-
`In accordance with the present
`pounds are provided which are inhibitors of MTP and have
`the structure
`
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`(wherein alkyl has at least 2 carbons, preferably at least
`
`4 of 162
`
`PENN EX. 2216
`CFAD V. UPENN
`
`lPR2015-01836
`
`

`
`6,066,650
`
`5
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`
`Her. '1
`
`—:R“T7J
`
`R'2—Z2
`
`z
`
`IIer2
`
`R13X"""\\R14
`
`W
`
`R] is "'1 i"d"”"l'“T’° gm“? °“h“ ”””°“"“
`
`.
`u
`y
`'3
`Z1 and Z“ are the same or diflercnt and are independently
`a bond, 0, S,
`
`D
`
`s
`
`s
`
`t';(tL) ’—““—i.r’—trtt—-
`
`0
`
`alkyl 0
`
`2
`
`jC‘ or jg’.-lj,
`II
`I
`0
`on
`
`with the proviso that with respect to B, at least one of Z1 and
`Z3 will be other than a bond;
`
`R“ is a bond, alkylene, alkenylene or alkynylenc of up to
`10 carbon atoms, arylene (for example
`
`an
`
`65
`
`5 of 162
`
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`
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`
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`
`Q),
`
`or mixed arylcne-alkylcnc [For example
`
` ([?H2)i.
`
`)
`
`where n is 1 to 6;
`R17‘
`is hydrogen, alkyl, alkenyl, aryl, haloalkyl,
`trihaloalkyl,
`trihaloalkylalkyl, heteroaryl,
`heteroarylalkyl, arylalkyl, arylalkenyl, cycloalkyl,
`aryloxy,a1koxy,arylalleoxyorcycloalkylalkyl,withthe
`provisos that (1) when R13 is H, aryloxy, alkoxy or
`arylalkgxy, than Z3 is
`
`NH
`
`;
`
`(3
`||
`0
`
`)
`
`C
`
`N
`|
`3”i}’l 0
`
`C
`
`0
`
`10
`
`15
`
`20
`
`25
`
`and (2) when Z2 is a bond, R12 cannot be heteroaryl or
`hctcmarylalkyl;
`Z is a bond, 0, S, N-alkyl, N-aryl, or alkylene or alk-
`.
`,
`enylene ot from 1 to 5 carbon atoms,
`Lk 1
`R13 R14 R15
`d H h d
`(‘R16
`‘
`I
`’an
`h’
`’
`arm“ cpcn en Y Y mgcma Y’
`R10’ hal0alkyl' aryl’ cycloalkyl’ Cyclohctcroalkyl’
`alkcnyl’ alkynyl‘ hydroxy’ alkoxy’ ‘mm’ amino’ thio’
`“”‘,Y15”1E’“Y1l’
`anriyllsullgonyll’ alkylfhuij’ arlyllhlo’
`aminoear ony , a y car ony oxy, ary ear ony amino,
`_
`alkylcarbonylamino, arylalkyl, heteroaryl, 33
`heteroarylalkyl, or aryloxy;
`_
`_
`.
`R15“ and R1” are independently any of the R15 or R1”
`groups except hydmxy’ Hum’ amino or thin;
`1 ,
`or R is
`
`30
`
`40
`
`8
`R5 is alkyl, alkenyl, alkynyl, aryl, alkoxy, aryloxy,
`arylalkoxy, heteroaryl, arylalkyl, heteroarylalkyl,
`cycloalkyl, cycloheteroalkyl, heleroaryloxy,
`cycloalkylalkyl, polycyeloalkyl, polyeycloalkylalkyl,
`cycloalkenyl, cycloalkenylalkyl, polycycloalkenyl,
`polyeycloalkenylalkyl, heteroarylcarbonyl, amino,
`alkylamino, arylamino, heteroarylarnino,
`cycloalkyloxy, cycloalkylamino, all of the R5 substitu-
`ents and R“ substituents (set out hereinafter) being
`optionally substituted through available carbon atoms
`with 1, 2, 3 or 4 groups selected from hydrogen, halo,
`alkyl, haloalkyl, alkoxy, haloalkoxy, alkenyl, alkynyl,
`eyeloalkyl, cycloalkylalkyl, cycloheteroalkyl,
`eycloheteroalkylalkyl, aryl, heteroaryl, arylalkyl,
`3TYlC}’ClU3“<}’1» iifylalkfiflyl.
`iiryliilklmyl. NYIUXY.
`al')’10XYa1k)'L afylalkoxy» “FY1330: h°l°1'0"lr}’10X0s
`h0l0l'03l')’l‘3lk}’1» h0i0i'031')"1fl1k°T1Y1a
`hCl°1'0?11'3’10XYs
`h)’d1'0X)'a
`f1i1l'0. W300, af1'|if10, Sub-Still-1105 51111100
`(wherein the amino includes 1 or 2 substituents which
`are alkyl, aryl or heteroaryl, or any of the other aryl
`compounds mentioned in the definitions),
`thiol,
`alkylthio, arylthio, heteroarylthio, arylthioalkyl,
`alkylcarbonyl, arylcarbonyl, arylamlnocarhonyl,
`alkoxycarbonyl,
`aminocarbonyl,
`alkynylaminocarbonyl, alkylaminocarhonyl,
`filth;:£:[:::u:E:rbUn);l1’k alltlgrlfi grub 01:): Liirfg’
`ar )lcarbon lalihino yar lsulfiii
`1
`ar lsullin lalk l’
`y
`y
`’
`y i
`y ’
`y i
`Y
`. Y ’
`arylsulfonyl, alkylsulfonyl, arylsulfonylamino,
`.
`,
`,
`__
`_
`,
`heteroarylcarbonylamlno, heteroarylsulllnyl,
`heteroarylthio, heteroarylsulfonyl, or alkylsulfinyl.
`Where R5 is phenyl aryl heteroaryl or cycloall<yl' this
`group preferably in’cludi*:s an ortho hydrophohic,sub-
`slituent such as alkyl, haloalkyl (with up to 5 halo
`_.
`.1k
`h.1 .1k
`- h
`5 h 1
`__
`Ex]:-lpi;,r);’1:1)x;:xU};, a:y(1:i]k;’1).(y (Wu up 10
`aogmups)’
`,
`,
`6 -,
`’
`.
`.
`_
`’,
`.
`.
`_
`R is hydrogen or (. —(. alltyl or C. —(. alkenyl,
`7 -
`1
`4
`-1
`4
`R is alkyl, aryl or arylalkyl wherein alkyl-or the alkyl
`portion is optionally substituted with oxo, and
`
`R17
`
`—w=—<R13
`
`wherein p is 1 to 8 and R” and R” are each independently
`H, alkyl, alkenyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl,
`cycloalkyl or cycloalkylalkyl, at least one of R” and R13
`hcing Othcr lhan
`or R1 is
`
`45
`
`50
`
`.
`.
`.
`Em the Sarlm or dificrclijl and grcdndcpcnicmiy Scicctcd from
`ct_crl_:])a1[-1):‘ corrflalnlfig‘ _ (Fr hfflgg mmln 01:’
`H
`d H_
`Int. U lflg
`-‘OX1 CS‘ -0
`l C
`OITTIU. ae
`.,
`1,
`an
`1
`compounds, that is
`
`TR
`
`19
`
`<
`
`Ran
`R21
`
`W /0 __md
`R
`P N‘ 1,‘
`55 K)
`
`wherein R” is aryl or heteroaryl;
`R20 i5 aryl 01' h°l°1'0a1'y1§
`R21 is II, alkyl, aryl, alkylaryl, arylalkyl, aryloxy, an
`arylalko xy, hetero aryl, heteroary lalkyl,
`heteroarylalkoxy, cycloalkyl, eycloalkylalkyl or
`cycloalkylalkoxy;
`R2, R3, R4 are ii'1(lepen(lcntly hydrogen, halo, alkyl,
`alkenyl,alkoxy,aryloxy,ary1,arylalkyl,alkylmercapto, 65
`arylmercapto, eyeloalkyl, cycloalkylalkyl, heteroaryl,
`heteroarylalkyl, hydroxy or haloalkyl;
`
`including pharmaceutically acceptable salts thereof such
`as alkali metal salts such as lithium sodium or
`potassium, alkaline earth metal salts such as calcium or
`magnesium, as well as zinc or aluminum and other
`cations such as ammonium, choline, diethanolamine,
`ethylenediamine,
`t-butylamine,
`t-oclylamine,
`dehydroahietylamine, as well as pharmaeeutically
`acceptable anions such as chloride, bromide, iodide,
`tartrate, acetate, methanesulfonate, maleate, succinate,
`
`6 of 162
`
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`
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`
`6,066,650
`
`glutarate, and salts of naturally occurring amino acids
`such as arginine,
`lysine, alanine and the like, and
`prodrug esters thereof.
`
`‘
`_
`In the formula I corripounds, where X 1s(.II2 and R2, R3 5
`and R’: are each II, R1 will be other than 3,3-diphenylpropyl.
`In the formula III compounds, where one of R2, R3 and R"
`,
`.
`is 6—fluoro, and the others are H, R7 wtll be other than
`4-(2-riiethoxy)pheriyl.
`Thus, the compounds of formulae I and II of the invention
`encompass compounds of the structure
`
`10
`
`R2
`F /
`1<3—
`
`l
`
`0
`
`X
`
`Q CK
`
`.\'—R‘
`
`R-1
`
`S
`
`R
`
`I
`
`R2
`0
`T
`N
`/
`R O
`J’/
`1<3—
`|
`N
`
`-
`
`R4
`
`R3
`
`0
`
`[3
`
`I5
`
`I 3
`1
`
`1"
`
`..
`
`as
`
`30
`
`1
`
`\—R
`
`R,
`IL
`
`_,_-m-m‘m_lcd
`
`O
`
`0
`
`0
`
`N
`
`0
`
`I
`
`2
`h?R
`
`RI
`I
`\
`
`x
`
`"N.
`
`R3
`
`[4
`Ii‘
`R4
`
`i
`
`R
`
`R3
`rr /
`NT |
`I‘ Ix"
`R4
`0
`
`5
`
`R
`
`R”
`
`O
`
`JL
`
`N
`|61:
`
`~
`
`I
`R6
`
`1“
`
`d1
`'
`
`II“
`
`11,’
`
`Hh
`
`2
`R /
`(2
`1*“ "‘
`1
`R
`
`|
`
`R-‘
`
`1:2
`
`/
`
`1’?
`]\.,_
`1:
`4 “~
`
`1:2
`J4 I
`]\
`~s. \
`R:
`
`R3
`
`i
`
`-«3— I O s
`
`N
`
`N—R'
`
`tt]
`
`R
`
`R9
`
`0
`
`I
`
`R
`l
`N
`
`as
`
`'1”
`
`411
`
`0
`1!
`\
`125/ ||\‘N
`0
`|r
`R’
`
`0
`II
`Rs/fi“-N
`0
`l
`R6
`
`N—R'
`
`R‘
`I
`N
`
`"lb
`
`N
`
`Rm
`
`9
`
`R
`
`O
`
`N
`/ m
`R
`
`R9
`
`The compounds of formula III of the tnvention encompass
`eompou nds of the structure III"
`
`.
`
`.
`
`.
`
`4:
`
`N—R'
`
`[C
`
`O
`
`59
`
`55
`
`R3
`I,,\/
`“ET
`7"‘
`R4
`
`|
`
`R
`xx 1
`
`(CH 1/N
`2 m
`
`111*
`
`111*’
`
`4
`
`R
`
`9
`
`R
`
`55
`
`.
`.
`.
`.
`.
`.
`In addition, in accordance with the present invention, a
`method for preventing, inhibiting or treating atherosclerosis,
`panereatitis or obesity is provided, wherein a compound of
`
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`
`6,066,650
`
`12
`be fused to l or 2 aromatic rings as described for aryl, which
`include cyclopropyl, cyelobutyl, cyclopentyl, cyelohexyl,
`cycloheptyl, cyclooctyl, cyclodecyl and cyclododecyl,
`cyclohexenyl,
`
`11
`formula I, Ii, II, IIi or III as defined hereinbefore wherein R'
`also includes arylmethyl, hetcroarylmethyl and cycloalkyl—
`methyl and Y also includes —(TII2—, is administered in an
`amount which decreases the activity of microsomal triglyc-
`eride transfer protein.
`Furthermore, in accordance with the present invention, a
`method is provided for lowering serum lipid levels, choles-
`terol andfor
`triglycerides, or
`inhibiting andfor treating
`hyperlipcmia, hyperlipidemia, hyperlipoproteinemia, hyper-
`cholesterolemia andfor hypertriglyceridemia, wherein a
`compound of formula I, Ii, II, IIi or III as defined herein-
`before wherein R‘
`also includes arylmethyl,
`heteroarylmethyl, and cycloalkylmethyl, and Y also includes
`—CH3—, is administered in an amount which decreases the
`activity of microsomal triglyceride transfer protein.
`
`10
`
`20
`
`30
`
`40
`
`DETAILED DESCRIPTION OF THE
`INVENTION
`
`The following definitions apply to the terms as used
`throughout this specification, unless otherwise limited in
`specific instances.
`The term “MTP” refers to a polypeptide or protein
`complex that (1) if obtained from an organism (e.g., cows,
`humans, etc.), can be isolated from the microsomal fraction
`of homogenized tissue; and (2) stimulates the transport of
`triglycerides, cholesterol esters, or phospholipids from syn-
`thetic phospholipid vesicles, membranes or lipoproteins to
`synthetic vesicles, membranes, or lipoproteins and which is
`distinct from the cholesterol ester transfer protein [Drayna et
`al., Nature 327, 632-634 (l987)] which may have similar
`catalytic properties. Ilowever,
`the MTP molecules of the
`present invention do not necessarily need to be catalytically
`active. For example, catalytically inactive MTP or fragments
`thereof may be useful in raising antibodies to the protein.
`The phrase “stabilizing” atherosclerosis as used in the
`present application refers to slowing down the development
`of andfor inhibiting the formation of new atherosclerotic
`lesions.
`
`any of which groups may be optionally substituted with 1 to
`4 substituents such as halogen, alkyl, alkoxy, hydroxy, aryl,
`aryloxy, arylalkyl, cycloalkyl, alkylamido, alkanoylamino,
`oxo, acyl, arylcarbonylamino, amino, nitro, cyano,
`thiol
`andfor alkylthio, as well as any of the other substituents as
`defined for R5 or R5.
`The term “cycloalkenyl” as employed herein alone or as
`part of another group refers to cyclic hydrocarbons contain-
`ing 5 to 20 carbons, preferably 6 to 12 carbons and l or 2
`double bonds. Exemplary cycloalkenyl groups include
`cyclopentenyl, cyelohexenyl, cyeloheptenyl, cyclooctenyl,
`cyclohexadienyl, and cycloheptadicnyl, which may be
`optionally substituted as defined for cycloalkyl.
`'lhe term “polycycloalkyl” as employed herein alone or as
`part of another group refers to a bridged multicyclic group
`containing 5 to 20 carbons and containing 0 lo 3 bridges,
`preferably 6 to 12 carbons and 1 or 2 bridges. Exemplary
`polycycloalkyl groups include [3.3.0]-bicyclooctanyl,
`adamantanyl,
`[2.2.1]—bicyc1oheptanyl,
`[2.2.2]—
`bicyclooctanyl and the like and may be optionally substi-
`tuted as defined for cycloalkyl.
`The term “polycycloalkenyl" as employed herein alone or
`as part of another group refers to bridged multicyclic group
`containing 5 to 20 carbons and containing 0 to 3 bridges and
`containing 1 or 2 double bonds, preferably 6 to l2 carbons
`The phrase “causing the regression of" atherosclerosis as
`and 1 or 2 bridges. Exemplary polycycloalkyl groups
`used in the present application refers to reducing andfor
`include [3.3.{l]-bicyclooctenyl, [2.2.l]-bicycloheptenyl,
`eliminating atherosclerotic lesions.
`[2.2.2]—bicyclooctcnyl and the like and may be optionally
`Unless otherwise indicated,
`the term “lower alkyl”,
`substituted as defined for cycloalkyl.
`“alkyl” or “alk” as employed herein alone or as part of
`The term “aryl” or “A1” as employed herein alone or as
`another group includes both straight and branched chain
`45 part of another group refers to monocyclic and bicyclic
`_
`_
`aromatic groups containing 6 to 10 carbons in Inn ring
`hydrocarbons, eontarningl to 40 carbons,preferal)lyl to 20
`not-non (Such as phcny] 01- nap}-nhyl) and may optionally
`carbons, more preferably 1
`to 12 carbons, in the normal
`include one to three additional rings fused to Ar (such as
`Chains-“Ch 35 methyl» clhlils PmP)’1ai50PmP)’l- b11lY]al'b“1Yla
`aryl, cycloalkyl, heteroaryl or cycloheteroalkyl rings) and
`lwbulylapcnlyla hcxyls isohcxyla hcplyl»4»4‘dimclhYlP'3mYL
`0‘3l)’1:2:3:4‘l1'im°lhY1l3°nlY1:HOHYLdCCY1:Un(l9CY1:d0(l'3CY1» 50 may be optionally substituted through available carbon
`the various branched chain isomers thereof, and the like as
`atoms will-, 1, 2, 3 or 4 grflups Selccwd from hydrogen, halo,
`“'9'” 35 Such QTOUP5 including 1 lo 4 5‘-1b5lilU"~5m5 Such 35
`haloalkyl, alkyl, haloalkyl, alkoxy, haloalkoxy, alkenyl,
`halos ff” cxamplc Fe B13 C1 01' I 01' CH3: “ROXY: 31'Y1: afyloxys
`trifluoromethyl, trifiuoromethoxy, alkynyl, cycloalkylalkyl,
`aryliafyl)
`01' diaryls arylalk-Y1:
`a1'Y13lkYl0XYs alkcnyla
`cycloheteroalkyl, cycloheteroalkylalkyl, aryl, heteroaryl,
`Cycloalkyls Cycloalkylalkyls CYCIO9-lkylalkyloxyy 331100» 55 arylalkyl, aryloxy, aryloxyalkyl, arylalkoxy, arylthio,
`hydroxys acyla hctcroaryla hctcroaryloxy» hclemafylal-kyls
`arylazo, heteroarylalkyl, heteroarylalkenyl,
`h°l°"031'Y1alk0XY» '<1rY1°XYa1k}’1»
`ar}’1°XYaFY1» 31kY1amid0»
`heteroarylhcteroaryl, hcteroaryloxy, hydroxy, nitro, cyano,
`alkafloylaminoa 31'ylcarbonylamlno» HMO» cY'<m0a
`‘M01:
`amino, substituted amino wherein the amino includes 1 or 2
`haloalkyla Irlhaloalkyl anwor alkyllhlos as “fcll 35 any Qflhc
`substituents (which are alkyl, aryl or any of the other aryl
`Ullwf 5UbS1i111'~‘>1'115 3-5 dcrlflfll {O1 R5 and R°«
`60 compounds mentioned in the definitions), thiol, alkylthio,
`Unless otherwise indicated,
`the term "Cycloalkyl” as
`arylthio, heteroarylthio, arylthioalkyl, alkoxyarylthio,
`employed herein alone or as part of another group includes
`alkylcarbonyl, arylcarbonyl, alkylaminocarbonyl,
`saturated or partially unsaturated (containing 1 or 2 double
`arylaminocarbonyl, alkoxycarbonyl, aminocarbonyl,
`bonds) cyclic hydrocarbon groups containing 1 to 3 rings,
`alkylcarbonyloxy, arylcarbonyloxy, alkylcarbonylamino,
`including monoeyclicalkyl, bieyelicalkyl and tricyelicalkyl, 65 arylcarbonylamino, arylsulfinyl, arylsulfinylalkyl, arylsulfo-
`containing a total of 3 to 20 carbons forming the rings,
`nylamino or arylsulfonaminocarbonyl, or any of the sub-
`prefcrably 4 to 12 carbons, forming the ring and which may
`stituents as defined for the R5 or R5 groups set out above.
`
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`13
`'lhe term “aralkyl", “aryl-alkyl” or “aryllower alkyl” as
`used herein alone or as part of another group refers to alkyl
`groups as discussed above having an aryl suhstituent, such
`as benzyl or phenethyl, or naphthylpropyl, or an aryl as
`defined above.
`
`The term "lower alkoxy”, “alkoxy”, "aryloxy” or
`“aralkoxy” as employed herein alone or as part of another
`group includes any of the above alkyl, aralkyl or aryl groups
`linked to an oxygen atom.
`'lhe tenn “amino” as employed herein alone or as part of
`another group may optionally be substituted with one or two
`substituents such as alkyl andfor aryl,
`The term “lower alkylthio”, alkylthio”, “arylthio” or
`“aralkylthio” as employed herein alone or as part of another
`group includes any of the above alkyl, aralkyl or aryl groups
`linked to a sulfur atom.
`
`The term “lower alkylamino”, “