United States Patent [19]
`Biller et al.
`
`111111
`
`11111111111111~111111111111111111111111111111111111111
`US005712279A
`5,712,279
`Patent Number:
`Jan. 27, 1998
`Date of Patent:
`
`[11]
`
`[45]
`
`[54]
`
`INHffiiTORS OF MICROSOMAL
`TRIGLYCERIDE TRANSFER PROTEIN AND
`METHOD
`
`[75] Inventors: Scott A. Biller, Hopewell; John K.
`Dickson, Eastampton, both of N.J.; R.
`Michael Lawrence, Yardley, Pa.; David
`R. Magnin, Hamilton; Michael A.
`Poss, Lawrenceville, both of N.J.;
`Jeffrey A. Robl, Newtown, Pa.;
`Richard B. Sulsky, Franklin Park;
`Joseph A. Tino, Lawrenceville, both of
`N.J.
`
`[73] Assignee: Bristol-Myers Squibb Company,
`Princeton, N.J.
`
`[21] Appl. No.: 548,811
`
`[22] Filed:
`
`Jan. 11, 1996
`
`Related U.S. Application Data
`
`[51]
`
`[63] Continuation-in-part of Ser. No. 472,067, Jun. 6, 1995,
`which is a continuation-in-part of Ser. No. 391 ,901, Feb. 21,
`1995, abandoned.
`Int. c1.6
`...................... com 211/98; C07D 409/06;
`C07D 405/06; A61K 31/445
`[52] U.S. Cl ........................... 514/252; 546/208; 546/202;
`546/198; 546/193; 546/189; 546/201; 546/194;
`546/196; 546/187; 546/244; 546/212; 546/214;
`546/200; 546/224; 546/203; 546/205; 546/199;
`546/213; 546/141; 514/325; 514/324; 514/321;
`514/318; 514/316; 514/259; 514/235.5;
`514/255; 514/228.8; 514/323; 514/320;
`514/232.8; 514/309; 514/329; 544/287;
`544/130; 544/360; 544/88; 544/405; 544/364;
`544/238; 544/391; 544/399
`[58] Field of Search ..................................... 546/208, 202,
`546/198, 193, 189, 201; 514/325, 324,
`321, 318, 316, 259, 235.5, 255, 228.8.
`323,252; 544/287,130,360,88,405
`
`[56]
`
`References Cited
`
`U.S. PATENT DOCUMENfS
`
`3,910,931 10/1975 Cavalla et al ........................... 546/206
`9/1981 Bengtsson et al ...................... 424/267
`4,289,781
`4,576,940
`3/1986 Tahara et al ............................ 5141212
`4,581,355
`4/1986 Tahara et al ............................ 5141212
`4,607,042
`8/1986 Pierce ...................................... 514/323
`5,032,598
`7/1991 Baldwin et al ......................... 514/318
`7/1992 Pan et al ................................. 514/460
`5,130,333
`2/1993 Peglion et al ........................... 514/319
`5,189,045
`5,215,989
`6/1993 Baldwin et al ......................... 514/252
`6/1996 Masuda et al .......................... 514/255
`5,527,801
`
`FOREIGN PATENT DOCUMENTS
`
`3/1994 European Pat. Off ..
`0584446A2
`3/1995 European Pat. Off ..
`0643057A1
`4/1993 WIPO .
`W09305778
`W094/40640 12/1996 WIPO .
`
`OTHER PUBLICATIONS
`
`Wetterau, J. and Zilversmit, D.B., J. Biol. Chern. 259,
`10863-10866 (1984), "1 Triglyceride and Cholesteryl Ester
`Transfer Protein Associated with liver Microsomes".
`
`Wetterau, J., Grant Application entitled: "Intracellular Trig(cid:173)
`lyceride Transport and Metabolism". Presentation Materials,
`Aspen Bile Acid/Cholesterol Conference, Aug. 15, 1992.
`Wetterau, J. R., et al., Science, vol. 258, 999-1001, Nov. 6,
`1992, "Absence of Microsomal Triglyceride Transfer Pro(cid:173)
`tein in Individuals with Abetalipoproteinemia".
`Archibald, J. L., et al., Journal of Medicinal Chemistry, vol.
`14, No. 11, pp. 1054-1059.
`Cortizo, L. et al., J. Med. Chern., 34, pp. 2242-2247. 1991.
`Hall, L H. et al., Pharmaceutical Research, vol. 9, No. 10, pp.
`1324-1329, 1992.
`Hall, L H., et al., Pharmacological Research Communica(cid:173)
`tions, vol. 19, No. 12, pp. 839-858, 1987.
`Murthy et al., Bur. J. Med. Chem.--Chim. Ther., vol. 20, No.
`6, pp. 547-550, 1985.
`Derwent Abstract No. 93-117225/14.
`Bulleid & Freedman, Nature 335, 649-651 (1988). "Defec(cid:173)
`tive co-translational forrilation of disulphide bonds in pro(cid:173)
`tein disulphideisomerase-deficient microsomes".
`Koivu et al., J. Bioi. Chern. 262, 6447-6449 (1987). "A
`Single Polypeptide Acts Both as the ~ Subunit of Prolyl
`4-Hydroxylase and as a Protein Disulfide-Isomerase".
`Kane & Havel in the Metabolic Basis of Inherited Disease,
`Sixth Edition, 1139-1164 (1989). "Disorders of the Biogen(cid:173)
`esis and Secretion of lipoproteins Containing The B Apo(cid:173)
`lipoproteins".
`Schaerer et al., Clin. Chern. 34, B9-B12 (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
`eDNA".
`
`(List continued on next page.)
`
`Primary Emminer-Mukund J. Shah
`Assistant Examiner-KID.g Lit Wong
`Attorney, Agent, or Firm-Burton Rodney
`
`[57]
`
`ABSTRACT
`
`Compounds are provided which inhibit microsomal triglyc(cid:173)
`eride transfer protein and thus are useful for lowering serum
`lipids and treating atherosclerosis and related diseases. The
`compounds have the structure
`
`0
`II
`C-NH-CHz-CF3
`
`(CHz).-NJ-N -c-Rs
`
`H
`
`II
`0
`
`wherein z, X 1
`
`' X 2
`
`' X and R 5 are as defined herein.
`
`19 Claims, No Drawings
`
`CFAD Ex. 1019 (1 of 98)
`
`

`
`5,712,279
`Page 2
`
`UfliER PUBLICATIONS
`
`Pihlajaniemi et al., EMBO J. 6, 643-649 (1987). "Molecular
`cloning of the ]>-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,
`1485-1492 (1987). "Sequence of Membrane-Associated
`Thyroid Hormone Binding Protein From Bovine Liver: Its
`Identity with Protein Disulphide Isomerase".
`Edman et al., Nature 317, 267-270 (1985). "Sequence of
`protein disulphide isomerase and implications of its rela(cid:173)
`tionship to thioredoxin".
`Kao et al., Connective Tissue Research 18, 157-174 (1988).
`"Isolation of eDNA Clones and Genomic DNA Dones of
`~ubunit of Chicken Prolyl4-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(cid:173)
`ide Transfer Protein".
`
`Morton, R.E. et al., J. Biol. Chern.. 256, 1992-1995 (1981).
`"A Plasma Inhibitor of Triglyceride and Chloesteryl Ester
`Transfer Activities".
`
`Wetterau, J. et al., Biochem. 30, 4406-4412 (1991): "Struc(cid:173)
`tural Properties of the Microsomal Triglyceride-Transfer
`Protein Complex".
`
`Wetterau, J. et al., J. Biol. Chern.. 265, 9800-9807 (1990).
`"Protein Disulfide Isomerase Is a Component of the
`Microsomal Triglyceride Transfer Protein Complex".
`
`Wetterau, J. and Zilversrnit, D.B., Chern. and Phys. of Lipids
`38, 205-22 (1985). "Purification and Characterization of
`Microsomal Triglyceride and Cholesteryl Ester Transfer
`Protein From Bovine Liver Microsomes".
`
`Wetterau, J. and Zilversrnit, D.B., Biochimica et Biophysica
`Acta 875, 610-617 (1986). "Localization of intracellular
`triacylglycerol and cholesteryl ester transfer activity in rat
`tissues".
`
`CFAD Ex. 1019 (2 of 98)
`
`

`
`5,712,279
`
`1
`INHffiiTORS OF MICROSOMAL
`TRIGLYCERIDE TRANSFER PROTEIN AND
`METHOD
`
`CROSS-REFERENCE TO RELATED
`APPLICATIONS
`
`This is a continuation-in-part of application Ser. No.
`472,067 filed Jun. 6, 1995, which is a continutation-in-part
`of application Ser. No. 391,901 filed Feb. 21, 1995, now
`abandoned each of which is hereby incorporated by refer(cid:173)
`ence.
`
`FIELD OF THE INVENTION
`
`This invention relates to novel compounds which inhibit
`microsomal triglyceride transfer protein, and to methods for
`decreasing serum lipids and treating atherosclerosis employ(cid:173)
`ing such compounds.
`
`BACKGROUND OF THE INVENTION
`
`The microsomal triglyceride transfer protein (MTP) cata(cid:173)
`lyzes the transport of triglyceride (I'G), cholesteryl ester
`(CE), and phosphatidylcholine (PC) between small unila(cid:173)
`mellar vesicles (SUV). Wetterau & Zilversmit, Chem. Phys.
`lipids 38,205-22 (1985). When transfer rates are expressed
`as the percent of the donor lipid transferred per time, MIP
`expresses a distinct preference for neutral lipid transport
`(I'G and CE), relative to phospholipid transport. The protein
`from bovine liver has been isolated and characterized.
`Wetterau & Zilversmit, Chem. Phys. lipids 38, 205-22
`(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 MIP 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 40
`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
`MIP, respectively.
`Characterization of the 58,000 molecular weight compo- 45
`nent of bovine MIP indicates that it is the previously
`characterized multifunctional protein, protein disulfide
`isomerase (PDI). Wetterau et al., J. Bioi. Chem. 265, 9800-7
`(1990). The presence of PDI in the transfer protein is
`supported by evidence showing that (1) the amino terminal 50
`25 amino acids of the bovine 58,000 kDa component of
`MIP is identical to that of bovine PDI, and (2) disulfide
`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 55
`which by itself has no TG transfer activity, were able to
`immunoprecipitate bovine TG transfer activity from a solu(cid:173)
`tion containing purified bovine MIP.
`PDI normally plays a role in the folding and assembly of
`newly synthesized disulfide bonded proteins within the 60
`lumen of the endoplasmic reticulum. Bulleid & Freedman,
`Nature 335, 649-51 (1988). It catalyzes the proper pairing
`of cysteine residues into disulfide bonds, thus catalyzing the
`proper folding of disulfide bonded proteins. In addition, PDI
`has been reported to be identical to the beta subunit of 65
`human prolyl 4-hydroxylase. Koivu et al., J. Bioi. Chem.
`262, 6447-9 (1987). The role of PDI in the bovine transfer
`
`2
`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 HCl), a chaotropic
`5 agent (sodium perchlorate), or a nondenaturing detergent
`(octyl glucoside) results in a loss of transfer activity. Wet(cid:173)
`terau et al., Biochemistry 30, 9728-35 (1991). Isolated
`bovine PDI has no apparent lipid transfer activity, suggest(cid:173)
`ing that either the 88 kDa polypeptide is the transfer protein
`10 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. Acta 875,610-7 (1986). Lipid transfer activity was
`found in liver and intestine. Little or no transfer activity was
`15 found in plasma, brain, heart, or kidney. Within the liver,
`MIP 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
`20 characterized by a virtual absence of plasma lipoproteins
`which contain apolipoprotein B (apoB). Kane & Havel in
`The Metabolic Basis of Inherited Disease, Sixth edition,
`1139-64 (1989). Plasma TG levels may be as low as a few
`mgldL, and they fail to rise after fat ingestion. Plasma
`25 cholesterol levels are often only 20-45 mgldL. These abnor(cid:173)
`malities are the result of a genetic defect in the assembly
`and/or secretion of very low density lipoproteins (VLDL) in
`the liver and chylomicrons in the intestine. The molecular
`basis for this defect has not been previously determined. In
`30 subjects examined, triglyceride, phospholipid, and choles(cid:173)
`terol synthesis appear normal. At autopsy, subjects are free
`of atherosclerosis. Schaefer et al., Clin. Chem. 34, B9-12
`(1988). A link between the apoB gene and abetalipopro(cid:173)
`teinernia has been excluded in several families. Talmud et
`35 al., J. Clin. Invest. 82, 1803-6 (1988) and Huang et al., Am.
`J. Hum. Genet. 46, 1141-8 (1990).
`Subjects with abet alipoproteinemia are afilicted with
`numerous maladies. Kate & Havel, supra. Subjects have fat
`malabsorption and TG accumulation in their enterocytes and
`hepatocytes. Due to the absence of TG-rich plasma
`lipoproteins, there is a defect in the transport of fat-soluble
`vitamins such as vitamin E. 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 abet alipoproteinemic subjects includes dietary
`restriction of fat intake and dietary supplementation with
`vitamins A, E and K.
`In vitro, MfP 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 MfP
`have led to speculation that it plays a role in the assembly of
`plasma lipoproteins, as these are the sites of plasma lipo(cid:173)
`protein assembly. Wetterau & Zilversrnit, Biochem. Biophys.
`Acta 875,610-7 (1986). The ability ofMTPto catalyze the
`transport of TG between membranes is consistent with this
`hypothesis, and suggests that MfP may catalyze the trans(cid:173)
`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(cid:173)
`bly in HepG2 cells. Bostrom et al., J. Bioi. Chem. 263,
`4434-42 (1988). Their results suggest small precursor lipo(cid:173)
`proteins become larger with time. This would be consistent
`
`CFAD Ex. 1019 (3 of 98)
`
`

`
`5,712,279
`
`3
`with the addition or transfer of lipid molecules to nascent
`lipoproteins as they are assembled. MfP may play a role in
`this process. In support of this hypothesis, Howell and
`Palade, J. Cell Biol. 92, 833-45 (1982), isolated nascent
`lipoproteins from the hepatic Golgi fraction of rat liver. 5
`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 (1984), reported lipoproteins isolated from 10
`Golgi were consistently larger than those from the endo(cid:173)
`plasmic reticulum, again suggesting the assembly of lipo(cid:173)
`proteins is a progressive event.
`
`15
`
`Recent reports (Science, Vol. 258, page 999, 1992; D.
`Sh31p et. al., Nature, Vol. 365, page 65, 1993) demonstrate
`that the defect causing abet alipoproteinemia is in the MI'P
`gene, and as a result, the MfP protein. Individuals with abet
`alipoproteinemia have no MfP activity, as a result of muta- 20
`tions in the MI'P gene, some of which have been charac(cid:173)
`terized. These results indicate that MfP is required for the
`synthesis of apoB containing lipoproteins, such as VLDL,
`the precursor to LDL. It therefore follows that inhibitors of
`MI'Pwould inhibit the synthesis ofVLDL and LDL, thereby 25
`lowering VLDL levels, LDL levels, cholesterol levels, and
`triglyceride levels in animals and man.
`
`Canadian Patent Application No. 2,091,102 published 30
`Mar. 2, 1994 (corresponding to U.S. application Ser. No.
`117.362, filed Sep. 3, 1993 (file DC21b)) reports MI'P
`inhibitors which also block the production of apoB contain(cid:173)
`ing lipoproteins in a human hepatic cell line (HepG2 cells).
`This provides further support for the proposal that an MfP 35
`inhibitor would lower apoB containing lipoprotein and lipid
`levels in vivo. This Canadian patent application discloses a
`method for identifying the MI'P inhibitors
`
`4
`
`or
`
`or
`
`II
`
`ill
`
`where X is: CHR8
`
`,
`
`-CH-CH or
`I
`I
`R9
`RID
`
`-C=C-;
`I
`I
`R9 RID
`
`R8
`, R9 and R 10 are independently hydrogen, alkyl, alkenyl,
`alkynyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl,
`cycloalkyl, or cycloalkylalkyl;
`Yis -(CH2)m- or
`
`-e(cid:173)
`ll
`0
`
`40 where m is 2 or 3;
`R 1 is alkyl, alkenyl, alkynyl, aryl, heteroaryl, arylalkyl
`(wherein alkyl has at least 2 carbons), diarylalkyl,
`arylalkenyl, diarylalkenyl, arylalkynyl, diarylalkynyl,
`diarylalkylaryl, heteroarylalkyl (wherein alkyl has at least 2
`45 carbons), cycloalkyl, or cycloalkylalkyl (wherein alkyl has
`at least 2 carbons); all of the aforementioned R 1 groups
`being optionally substituted through available carbon atoms
`with 1, 2, or 3 groups selected from halo, haloalkyl, alkyl,
`alkenyl, alkoxy, aryloxy, aryl, arylalkyl, alkylmercapto,
`so arylmercapto, cycloalkyl, cycloalkylalkyl, heteroaryl,
`fluorenyl, heteroarylalkyl, hydroxy or oxo; or
`R 1 is a group of fue structure
`
`which has the name 2-[ 1-(3,3-diphenylpropyl)-4-
`piperidinyl]-2,3-dihydro-3-oxo-lH-isoindole hydrochloride
`and
`
`55
`
`60
`
`65
`
`which has the name 1-[3-(6-fluoro-1-tetralanyl)mefuyl]-4-
`0-rnethoxyphenyl piperazine
`
`EP 0643057 Al published Mar. 15, 1995, discloses MI'P
`inhibitors of the structure
`
`R 11 is a bond, alkylene, alkenylene or alkynylene of up to
`6 carbon atoms, arylene (for example
`
`CFAD Ex. 1019 (4 of 98)
`
`

`
`5
`
`-@-),
`
`or mixed arylene-alkylene (for example
`
`6
`arylthio, heteroarylthio, arylthioalkyl, alkylcarbonyl,
`arylcarbonyl, arylaminocarbonyl, alkoxycarbonyl,
`amino carbonyl,
`alkynylami nocarbonyl,
`alkyl amino carbonyl, alke nylamino carbonyl,
`5 alkylcarbonyloxy, arylcarbonyloxy, alkylcarbonylamino,
`arylcarbonylamino, arylsulfinyl, arylsulfinylalkyl,
`arylsulfonyl, alkylsulfonyl, arylsulfonylamino; with tlle pro(cid:173)
`viso that when R5 is CH3, R6 is not H; and where R5 is
`phenyl, the phenyl preferably includes an ortllo hydrophobic
`10 substituent such as alkyl, haloalkyl. aryl, aryloxy or aryla(cid:173)
`lkyl;
`R6 is hydrogen or CcC4 alkyl or CcC4 alkenyl;
`where n is 1 to 6;
`R7 is alkyl, aryl or arylalkyl wherein alkyl or the alkyl
`R 12 is hydrogen, alkyL alkenyl, aryl, heteroaryl, 15
`portion is optionally substituted with oxo; and
`haloalkyl, arylalkyl, arylalkenyl, cycloalkyl, aryloxy,
`alkoxy, arylalkoxy, heteroarylalkyl or cycloalkylalkyl;
`including pharmaceutically acceptable salts and anions
`Z is a bond, 0, S, N-alkyl, N-aryl, or alkylene or alk(cid:173)
`thereof.
`enylene of from 1 to 5 carbon atoms;
`, R3
`In the formula I compounds, where X is CH2 and R2
`R 13
`, R 14
`, R15
`, andR16 are independently hydrogen, alkyl,
`halo, haloalkyL aryl, cycloalkyl, cycloheteroalkyl, alkenyl, 20 and R4 are each H, R 1 will be other than 3,3-diphenylpropyl.
`alkynyl, hydroxy, alkoxy, nitro, amino, thio, alkylsulfonyl,
`, R3 and R4
`In the formula ill compounds, where one of R 2
`arylsulfonyl, alkylthio, arylthio, carboxy, aminocarbonyl,
`is 6-fluoro, and the others are H, R7 will be other tllan
`alkylcarbonyloxy, alkylcarbonylamino, arylalkyl,
`4-0-metlloxyphenyl.
`heteroaryl, heteroarylalkyl, or aryloxy;
`or R1 is
`U.S. application Ser. No. 472,067, filed Jun. 6, 1995 (file
`DC21e) discloses compounds of the structure
`
`5,712,279
`
`25
`
`wherein p is 1 to 8 and R 17 and R 18 are each independently
`H, alkyl, alkenyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl,
`cycloalkyl or cycloalkylalkyl, at least one of R17 and R 18
`being other than H;
`or R1 is
`
`35
`
`or
`
`30
`
`R20
`
`-R19-<
`R21
`
`40
`
`wherein
`R 19 is aryl or heteroaryl;
`R 20 is aryl or heteroaryl;
`R 21 is H, alkyl, aryl, alkylaryl, arylalkyl, aryloxy, 45
`arylalkoxy, heteroaryl, heteroarylalkyl, heteroarylalkoxy,
`cycloalkyl, cycloalkylalkyl or cycloalkylalkoxy;
`R2
`, R3
`, R4 are independently hydrogen, halo, alkyl,
`haloalkyl, alkenyl, alkoxy, aryloxy, aryl, arylalkyl,
`alkylmercapto, arylmercapto, cycloalkyl, cycloalkylalkyl, so
`heteroaryl, heteroarylalkyl, hydroxy or haloalkyl;
`R5 is alkyl of at least 2 carbons, alkenyl, alkynyl, aryl,
`heteroaryl, arylalkyl, heteroarylalkyl, cycloalkyl,
`cycloalkylalkyl, polycycloalkyl, polycycloalkylalkyl,
`cycloalkenyl, cycloalkenylalkyl, polycycloalkenyl, 55
`polycycloalkenylalkyl, heteroarylcarbonyl, all of the R5 and
`R6 substituents being optionally substituted through avail(cid:173)
`able carbon atoms with 1, 2, or 3 groups selected from
`hydrogen, halo, alkyl, haloalkyl, alkoxy, haloalkoxy,
`alkenyl, alkynyl, cycloalkyl, cycloalkylalkyl, 60
`cycloheteroalkyl, cycloheteroalkylalkyl, aryl, heteroaryl,
`arylalkyl, arylcycloalkyl, arylalkynyl, aryloxy, aryloxyalkyl,
`arylalkoxy, arylazo, heteroaryloxo, heteroarylalkyl,
`heteroarylalkenyl, heteroaryloxy, hydroxy, nitro, cyano,
`amino, substituted amino (wherein the amino includes 1 or 65
`2 substituents which are alkyl, or aryl or any of the other aryl
`compounds mentioned in the definitions), thiol, alkylthio,
`
`or
`
`or
`
`or
`
`R3
`
`R2
`
`0
`
`R4
`
`r
`
`R7
`N/
`
`y/N~
`
`CFAD Ex. 1019 (5 of 98)
`
`

`
`where Q is
`
`7
`
`0
`II
`-e-
`
`0
`II
`or -S-;
`II
`0
`
`-e(cid:173)
`ll
`0
`
`,
`
`-eH-eH
`I
`I
`R9
`RIO
`
`or -e=e-;
`I
`I
`R9 RIO
`
`, R9 and R 10 are independently hydrogen, alkyl, alkenyl,
`R8
`alkynyl. aryl. arylalkyl, heteroaryl, heteroarylalkyl,
`cycloalkyl, or cycloalkylalkyl;
`
`-e(cid:173)
`ll
`0
`
`wherein m is 2 or 3;
`
`5,712,279
`
`8
`-continued
`RlS
`
`5
`
`10
`
`15
`
`20
`
`25
`
`or
`
`or
`
`R1 is an indenyl-type group of the structure
`
`R 1 is alkyl, alkenyl, alkynyl, aryl, heteroaryl, arylalkyl
`wherein alkyl has at least 2 carbons, diarylalkyl, arylalkenyl, 30
`diarylalkenyl, arylalkynyl, diarylalkynyl, diarylalkylaryl,
`heteroarylalkyl wherein alkyl has at least 2 carbons,
`cycloalk.yl, or cycloalkylalkyl wherein alkyl has at least 2
`carbons, all optionally substituted through available carbon
`atoms with L 2, 3 or 4 groups selected from halo, haloalkyl. 35
`alkyl, alkenyl, alkoxy, aryloxy, aryl, arylalkyl,
`alkylmercapto, arylmercapto, cycloalkyl, cyclo-alkylalkyl,
`heteroaryl, fluorenyl, heteroarylalkyl, hydroxy or oxo;
`
`or R 1 is a fluorenyl-type group of the structure
`
`or
`
`or
`
`A
`
`40
`
`45
`
`50
`
`B
`
`55
`
`60
`
`65
`
`R16a
`
`(a= 2, 3 or4
`
`R16a
`
`RlS•
`
`or
`
`or
`
`Ron RH
`
`-R11-zl
`
`R12-z2
`RlS•
`
`(eH2)a
`
`R16a
`
`or
`
`e
`
`D
`
`E
`
`F
`
`G
`
`H
`
`CFAD Ex. 1019 (6 of 98)
`
`

`
`9
`Z1 and Z2 are the same or different and are independently
`a bond, 0, S,
`
`5,712,279
`
`~ ( fl ) ' -NH-w-, -f-w-, -w- or -J-,
`
`S,
`
`0
`
`2
`
`0
`
`alkyl 0
`
`0
`
`H
`
`OH
`
`with the proviso that with respect to B, at least one of Z 1 and
`Z2 will be other than a bond; R 11 is a bond, alkylene,
`alkenylene or alkynylene of up to 10 carbon atoms; arylene
`or mixed arylene-alkylene; R12 is hydrogen, alkyl, alkenyl,
`aryl, haloalkyl, trihaloalkyl, trihaloalkylalkyl, heteroaryl,
`heteroarylalkyl, arylalkyl, arylalkenyl, cycloalkyl, aryloxy,
`alkoxy, arylalkoxy or cycloalkylalkyl, with the provisos that
`(1) when R 12 isH, aryloxy, alkoxy or arylalkoxy, then Z2
`is
`
`-NH-C- -N-C-, -e-
`ll
`'
`I
`II
`II
`0
`alkyl 0
`0
`
`or a bond and
`(2) when z2 is a bond, R 12 cannot be heteroaryl or
`
`10
`
`10
`polycycloalkylalkyl, cycloalkenyl, cycloheteroalkyl,
`heteroaryloxy, cycloalkenylalkyl, polycycloalkenyl,
`polycycloalkenylalkyl, heteroarylcarbonyl, amino,
`alkylamine, acylamino, heteroarylamino, cycloalkyloxy,
`5 cycloalkylamino, all optionally substituted through avail(cid:173)
`able carbon atoms with 1, 2, 3 or 4 groups selected from
`hydrogen, halo, alkyl, haloalkyl, alkoxy, haloalkoxy,
`alkenyl, alkynyl, cycloalkyl, cycloalkylalkyl,
`cycloheteroalkyl, cycloheteroalkylalkyl, aryl, heteroaryl,
`arylalkyl, arylcycloalkyl, arylalkenyl, arylalkynyl, aryloxy,
`aryloxyalkyl, arylalkoxy, arylazo, heteroaryloxo,
`heteroarylalkyl, heteroarylalkenyl, heteroaryloxy, hydroxy,
`nitro, cyano, amino, substituted amino, thiol, alkylthio,
`arylthio, heteroarylthio, arylthioalkyl, alkylcarbonyl,
`arylcarbonyl, arylaminocarbonyl, alkoxycarbonyl,
`15 aminocarbonyl,
`alkynylaminocarbonyl,
`alkylamine carbonyl, alkenylamino carbonyl,
`alkylcarbonyloxy, arylcarbonyloxy, alkylcarbonylamino,
`arylcarbonylamino, arylsulfinyl, arylsulfinylalkyl,
`arylsulfonyl, alkylsulfonyl, arylsulfonylamino,
`20 heteroarylcarbonylamino, heteroarylsulfinyl, heteroarylthio,
`heteroarylsulfonyl, alkylsulfinyl;
`R6 is hydrogen or CcC4 alkyl or CcC4 alkenyl; all
`optionally substituted with 1, 2, 3 or 4 groups which may
`independently be any of the substituents listed in the defi-
`25 nition of R5 set out above;
`R7 is alkyl, aryl or arylalkyl wherein alkyl by itself or as
`part of arylalkyl is optionally substituted with oxo
`
`heteroarylalkyl;
`Z is bond, 0, S, N-alkyl, N-aryl, or alkylene or alken~lene
`from 1 to 5 carbon atoms; R 13
`, R1
`\ R15
`, and R1 are
`independently hydrogen, alkyl, halo, haloalkyl, aryl,
`cycloalkyl, cycloheteroalkyl, alkenyl, alkynyl, hydroxy,
`alkoxy, nitro, amino, thio, alkylsulfonyl, arylsulfonyl,
`alkylthio, arylthio, aminocarbonyl, alkylcarbonyloxy, 30
`arylcarbonylamino, alky1carbonylamino, arylalkyl,
`heteroaryl, heteroarylalkyl or aryloxy;
`5a and R 16a are independently hydrogen, alkyl, halo,
`R 1
`haloalkyl, aryl, cycloalkyl, cycloheteroalkyl, alkenyl,
`alkynyl, alkoxy, alkylsulfonyl, arylsulfonyl, alkylthio, 35
`arylthio, aminocarbonyl, alkylcarbonyloxy,
`arylcarbonylamino, alkylcarbonylamino, arylalkyl,
`heteroaryl, heteroarylalkyl, or aryloxy;
`or R 1 is a group of the structure
`
`40
`
`and
`
`wherein pis 1 to 8 and R 17 and R18 are each independently 45
`H, alkyl, alkenyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl,
`cycloalkyl or cycloalkylalkyl at least one of R17 and R18
`being other than H;
`or R 1 is a group of the structure
`
`50
`
`R21l
`
`-R19-<
`R21
`
`are the same or different and are independently selected from
`heteroaryl containing 5- or 6-ring members; and
`N-oxides
`
`55
`
`wherein
`R19 is aryl or heteroaryl;
`R20 is aryl or heteroaryl;
`R 21 is H, alkyl, aryl, alkylaryl, arylalkyl, aryloxy,
`arylalkoxy, heteroaryl, heteroarylalkyl, heteroarylalkoxy,
`cycloalkyl, cycloalkylalkyl or cycloalkylalkoxy;
`, R4 are independently hydrogen, halo, alkyl,
`, R3
`R2
`alkenyl, alkoxy, aryloxy, aryl, arylalkyl, alkylmercapto,
`arylmercapto, cycloalkyl, cycloalkylalkyl, heteroaryl,
`heteroarylalkyl, hydroxy or haloalkyl;
`R5 is independently alkyl, alkenyl, alkynyl, aryl, alkoxy, 65
`aryloxy, arylalkoxy, heteroaryl, arylalkyl, heteroarylalkyl,
`cycloalkyl, cycloalkylalkyl, polycycloalkyl,
`
`thereof; and
`pharmaceutically acceptable salts thereof;
`with the provisos that where in the first formula X is CH2 ,
`and R2
`, R3 and R4 are each H, then R 1 will be other than
`60 3,3-diyhenylpropyl, and in the fifth formula, where one of
`R2
`, R and R4 is 6-fluoro, and the others are H, R7 will be
`other than 4-(2-methoxyphenyl).
`
`SUMMARY OF THE INVENTION
`
`In accordance with the present invention, novel com(cid:173)
`pounds are provided which are inhibitors of MTP and have
`the structure
`
`CFAD Ex. 1019 (7 of 98)
`
`

`
`11
`
`5,712,279
`
`12
`-continued
`
`0
`II H
`C-N-CH2-CF3
`
`(CHz).-NJ-N-C-R5
`II
`H
`0
`
`5
`
`10
`
`15
`
`wherein Z is a bond, 0 or S;
`X 1 and X2 are independently H or halo, preferably F;
`x is an integer from 2 to 6, preferably from 3 to 5, and
`(CH2).. may be optionally substituted with 1, 2 or 3 sub(cid:173)
`stituents which are the same or different and are alkyl or 20
`halo; and
`R5 is heteroaryl, aryl, heterocycloalkyl or cycloalkyl, each
`R5 group being optionally substituted with 1, 2, 3 or 4
`substituents which may be the same or different as defined
`hereinafter; and
`including piperidine N-oxides of the formula I compound,
`that is
`
`25
`
`cr OH ,
`
`#'
`
`CH~CH3
`
`30
`
`N-N
`H
`
`35
`
`and
`including pharmaceutically acceptable salts thereof such
`as alkali met al salts such as lithium sodium or potassium,
`alkaline earth met al salts such as calcium or magnesium, as
`well as zinc or aluminum and other cations such as
`ammonium, choline, diethanolarnine, ethylenediamine, 40
`t-butyl-amine, t-octylarnine, dehydroabietylarnine, as well
`as pharmaceutically acceptable anions such as chloride,
`bromide, iodide, tartrate, acetate, methanesulfonate,
`maleate, succinate, glutarate, and salts of naturally occurring
`amino acids such as arginine, lysine, alanine and the like, 45
`and prodrug esters thereof.
`The R5 group may be substituted with 1, 2, 3 or 4
`substituents, including
`(1) halogen such as Cl, F, CF3 , and I,
`(2) heteroaryl, including monocyclic or bicyclic ring
`systems, which includes 1, 2 or 3 heteroatoms which
`are S, Nand/or 0, and which includes from 2 to 10
`carbons in the ring or ring system, such as
`
`50
`
`A
`~~'
`0 N
`~N l );
`
`N
`
`55
`
`(3) heteroarylalkyl wherein heteroaryl is as defined above
`such as
`
`65
`
`( 4) cycloheteroalkyl which includes 1, 2 or 3 hetero atoms
`which are N, S or 0 in a monocyclic or bicyclic ring
`system such
`
`CFAD Ex. 1019 (8 of 98)
`
`

`
`5,712,279
`
`13
`
`I
`
`I
`
`c·) c·) 0· U"
`
`I
`
`0
`
`I
`
`N
`I
`CH3
`
`(5) alkyl;
`(6) aryl such as phenyl, phenyl substituted with (a) halo,
`(b) alkyl, (c) CF30, (d) alkoxy (e)
`
`(f) CF3 , or (g) phenyl;
`(7) alkylamino such as
`
`(8) alkyl (aryl) amino such as -N(CH3)C6Hs;
`(9) alkythio such as ---S- (CH2)pCF3,
`
`---S-alkyl,
`
`OCH3 ;
`(11) cycloalkyl such as cyclohexyl;
`(12) aryloxy such as
`
`(13) amino;
`(14) acylamino such as
`
`(15) arylthio such as
`
`14
`
`5
`
`10
`
`15
`
`20
`
`25
`
`30
`
`35
`
`40
`
`NC
`
`-s-@-~ -s-@,
`
`(16) acyl such as (a) alkanoyl, such as
`
`(b) alkoxycarbonyl, such as
`
`0
`II
`-C-0-CFJ,
`
`(c) aroyl, such as
`
`0 0~
`-~-C61Is, -~---&-F,
`
`(d) heteroarylaminocarbonyl, such as
`
`45 (e) arylalkyloxycarbonyl, such as
`
`(17) arylthioalkyl, such as -CH2---S-C6Hs;
`(18) heteroarylamino, such as
`
`-: --\;0": -N(CH,)-CH,-Q,
`
`(19) arylalkyloxy, such as
`
`50
`
`55
`
`60
`
`65
`
`CFAD Ex. 1019 (9 of 98)
`
`

`
`15
`(20) heteroarylthio, such as
`
`5,712,279
`
`16
`-continued
`
`(21) heteroaryloxy, such as
`
`and
`
`(22) arylsulfinyl, such as
`
`5
`
`10
`
`0
`II H
`C-N -CH;!CF3
`
`(CH2)x-NJ-N-C-Rs.
`II
`H
`0
`
`15
`
`In addition, in accordance with the present invention, a
`method for preventing, inhibiting or treating atherosclerosis,
`pancreatitis or obesity is provided, wherein a compound of
`formula I as defined hereinbefore is administered in an
`amount which decreases the activity of microsomal triglyc-
`20 eride transfer protein.
`Furthermore, in accordance with the present invention, a
`method is provided for lowering serum lipid levels, choles(cid:173)
`terol and/or triglycerides, or inhibiting and/or treating
`hyperlipemia, hyperlipidemia, hyperlipoproteinemia, hyper-
`25 cholesterolemia and/or hypertriglyceridemia, wherein a
`compound of formula I as defined hereinbefore is adminis(cid:173)
`tered in an amount which decreases the activity of microso(cid:173)
`mal triglyceride transfer protein.
`
`Thus, the compounds of formula I of the invention 30
`encompass compounds of the structure
`
`I'
`
`J'>
`
`0
`II H
`C-N -CH;!CF3
`
`(CH,k-N J -q -R ' ,
`
`0
`II H
`C-N -CH:zCFJ
`
`(CHz),-NJ-N-C-RS; and
`II
`H
`0
`
`35
`
`DErAILED DESCRIPTION OF THE
`INVENTION
`The following definitions apply to the terms as used
`throughout this specification, unless otherwise limited in
`specific instances.
`The term "MfP" 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
`40 triglycerides, cholesterol esters, or phospholipids from syn(cid:173)
`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 (1987)] which may have similar
`45 catalytic properties. However, the MfP molecules of the
`present invention do not necessarily need to be catalytically
`active. For example, catalytically inactive MfP or fragments
`thereof may be useful in raising antibodies to the protein.
`The phrase "stabilizing" atherosclerosis as used in the
`so present application refers to slowing down the development
`of and/or inhibiting the formation of new atherosclerotic
`lesions.
`The phrase "causing the regression of' atherosclerosis as
`used in the present application refers to reducing and/or
`55 eliminating atherosclerotic lesions.
`Unless otherwise indicated, the term "lower alkyl",
`"alkyl" or "alk" as employed herein alone or as part of
`another group includes both straight and branched chain
`hydrocarbons, containing 1 to 40 carbons, preferably 1 to 20
`60 carbons, more preferably 1 to 12 carbons, in the normal
`chain, such as methyl, ethyl, propyl, isopropyl, butyl,
`t-butyl, isobutyl, pentyl, hexyl, isohexyl, heptyl, 4,4-
`
`65
`
`CFAD