AU753520B2 - Cyclic phosphonate ester inhibitors of microsomal triglyceride transfer protein and method - Google Patents

Description

WO 99/21564 PCT/US98/21750 CYCLIC PHOSPHONATE ESTER INHIBITORS OF MICROSOMAL TRIGLYCERIDE TRANSFER PROTEIN AND METHOD Field of the Invention This invention relates to novel cyclic phosphonate esters which inhibit microsomal triglyceride transfer protein, and to methods for decreasing serum lipids and treating atherosclerosis employing such compounds.

Background of the Invention The microsomal triglyceride transfer protein (MTP) catalyzes the transport of triglyceride cholesteryl ester and phosphatidylcholine (PC) between small unilamellar 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, MTP expresses a distinct preference for neutral lipid transport (TG 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 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 component of bovine MTP indicates that it is the previously characterized multifunctional protein, protein disulfide isomerase (PDI). Wetterau et al., J. Biol. Chem. 265, 1 WO 99/21564 PCT/US98/21750 9800-7 (1990). The presence of PDI in the transfer protein is supported by evidence showing that the amino terminal 25 amino acids of the bovine 58,000 kDa component of MTP 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 which by itself has no TG transfer activity, were able to immunoprecipitate bovine TG transfer activity from a solution containing purified bovine MTP.

PDI normally plays a role in the folding and assembly of newly synthesized disulfide bonded proteins within the 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 human prolyl 4hydroxylase. Koivu et al., J. Biol. Chem. 262, 6447-9 (1987). The role of PDI 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 HC1), a chaotropic agent (sodium perchlorate), or a nondenaturing detergent (octyl glucoside) results in a loss of transfer activity.

Wetterau et al., Biochemistry 30, 9728-35 (1991). Isolated bovine PDI has no apparent lipid transfer activity, suggesting 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. Acta 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 2 WO 99/21564 PCT/US98/21750 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 Havel in The Metabolic Basis of Inherited Disease, Sixth edition, 1139-64 (1989). Plasma TG 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/dL.

These abnormalities 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 subjects examined, triglyceride, phospholipid, and cholesterol 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 abetalipoproteinemia has been excluded in several families.

Talmud et al., J. Clin. Invest. 82, 1803-6 (1988) and Huang et al., Am. J. Hum. Genet. 46, 1141-8 (1990).

Subjects with abetalipoproteinemia are afflicted with numerous maladies. Kane Havel, vide 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 abetalipoproteinemic 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 3 WO 99/21564 PCT/US98/21750 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 lipoprotein assembly. Wetterau Zilversmit, Biochem. Biophys. Acta 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 transport 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 assembly in HepG2 cells. Bostrom et al., J. Biol. Chem.

263, 4434-42 (1988). Their results suggest small precursor lipoproteins become larger with time. This would be consistent with the addition or transfer of lipid molecules to nascent lipoproteins as they are assembled. MTP 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. 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 Golgi were consistently larger than those from the endoplasmic reticulum, again suggesting the assembly of lipoproteins is a progressive event.

However, there is no direct evidence in the prior art demonstrating that MTP plays a role in lipid metabolism or the assembly of plasma lipoprotein.

Recent reports (Science, Vol. 258, page 999, 1992; D. 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 abetalipoproteinemia have no MTP activity, 4 WO 99/21564 PCT/US98/21750 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 VLDL, the precursor to LDL. It therefore follows that inhibitors of MTP would inhibit the synthesis of VLDL and LDL, thereby lowering VLDL levels, LDL levels, cholesterol levels, and triglyceride levels in animals and man.

Canadian Patent Application No. 2,091,102 published March 2, 1994 (corresponding to U.S. application Serial No.

117,362, filed September 3 1993 (file DC21b)) which is incorporated herein by reference), reports MTP inhibitors which also block the production of apoB containing 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

N

which has the name 2-[1-(3,3-diphenylpropyl)-4piperidinyl]-2, 3-dihydro-3-oxo-lH-isoindole hydrochloride and 0 F

OCH

3 which has the name 1-[ 3 -(6-fluoro-l-tetralanyl)-methyl]-4- 0-methoxyphenyl piperazine.

EP 0643057A1 published March 15, 1995, discloses MTP inhibitors of the structure 5 WO 99/21564 PCT/US98/21750

I

O

R

3 N- N-R 1 R4 or

II

0 A or

III

0

R

4

Y

where X is: CHR 8 -CH- c or I I I I Rs 9 io 9

R

0

R

8

R

9 and R 10 are independently hydrogen, alkyl, alkenyl, alkynyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, cycloalkyl, or cycloalkylalkyl; Y is -(CH2)m- or -Co 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 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, arylmercapto, cycloalkyl, cycloalkylalkyl, heteroaryl, fluorenyl, heteroarylalkyl, hydroxy or oxo; or

R

1 is a group of the structure 6 WO 99/21564 WO 9921564PCT/US98/21 750 R11z R 1 2

R

11 is a bond, alkylene, alkenylene or alkynylene of up to 6 carbon atoms, arylene (for example Q7 or mixed arylene-alkylene (for example where n is 1 to 6;

R

1 2 is hydrogen, alkyl, alkenyl, aryl, heteroaryl, haloalkyl, arylalkyl, arylalkenyl, cycloalkyl, aryloxy, alkoxy, arylalkoxy, heteroarylalkyl or cycloalkylalkyl; Z is a bond, 0, S, N-alkyl, N-aryl, or alkylene or alkenylene of from 1 to 5 carbon atoms;

R

1 3

R

1 4

R

1 5 arnd R 16 are independently hydrogen, alkyl, halo, haloalkyl, aryl, cycloalkyl, cycloheteroalkyl, alkenyl, alkynyl, hydroxy, alkoxy, nitro, amino, thio, alkylsulfonyl, arylsulfonyl, alkylthio, arylthio, carboxy, aminocarbonyl, alkylcarbonyloxy, alkylcarbonylanino, arylalkyl, heteroaryl, heteroarylalkyl, or aryloxy; or R 1 is

-I(C

2 )v -K wherein p is 1 to 8 and R 1 7 and R 18 are each independently H, alkyl, alkenyl, aryl, arylalkyJ., heteroaryl, heteroarylalkyl, cycloalkyl or cycloalkylalkyl, at least one of R 17 and R 1 8 being other than H; or R 1 is -7 WO 99/21564 WO 9921564PCT/US98/21 750

R

2 0 wherein R 1 9 is aryl or heteroaryl;

R

20 is aryl or heteroaryl;

R

2 1 is H, alkyl, aryl, alkylaryl, arylalkyl, -aryloxy, arylalkoxy, heteroaryl, heteroarylalkyl, heteroarylalkoxy, cycloalkyl, cycloalkylalkyl. or cycloalkylalkoxy;

R

2

R

3

R

4 are independently hydrogen, halo, alkyl, haloalkyl, alkenyl, alkoxy, aryloxy, aryl, arylalkyl, alkylmercapto, arylmercapto, cycloalkyl, cycloalkylalkyl, heteroaryl, heteroarylalkyl, hydroxy or haloalkyl;

R

5 is alkyl of at least 2 carbons, alkenyl, alkynyl, aryl, heteroaryl, arylalkyl, heteroarylalkyl, cycloalkyl, cycloalkylalkyl, polycycloalkyl, polycycloalkylalkyl, cycloalkenyl, cycloalkenylalkyl, polycycloalkenyl, polycycloalkenylalkyl, heteroarylcarbonyl, all of the R and R 6 substituents being optionally substituted through available carbon atoms with 1, 2, or 3 groups selected from hydrogen, halo, alkyl, haloalkyl, alkoxy, haloalkoxy, alkenyl, alkynyl, cycloalkyl, cycloalkylalkyl, 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 2 substituents which are alkyl, or aryl or any of the other aryl compounds mentioned in the definitions), thiol, alkylthio, arylthio, heteroarylthio, arylthioalkyl, alkylcarbonyl, arylcarbonyl, arylaminocarbonyl, alkoxycarbonyl, aminocarbonyl, alkynylaminocarbonyl, alkylaminocarbonyl, alkenylaminocarbonyl, alkylcarbonyloxy, arylcarbonyloxy, alkylcarbonylamino, arylcarbonylamino, arylsulfinyl, arylsulfinylalkyl, arylsulfonyl, alkylsulfonyl, arylsulfonylamino; with the proviso that when R 5 is CH 3

R

6 8- WO 99/21564 PCT/US98/21750 is not H; and where R 5 is phenyl, the phenyl preferably includes an ortho hydrophobic substituent such as alkyl, haloalkyl, aryl, aryloxy or arylalkyl;

R

6 is hydrogen or Ci-C 4 alkyl or CI-C 4 alkenyl;

R

7 is alkyl, aryl or arylalkyl wherein alkyl or the alkyl portion is optionally substituted with oxo; and including pharmaceutically acceptable salts and anions thereof or esters thereof.

In the formula I compounds, where X is CH 2 and R 2

R

3 and R 4 are each H, R 1 will be other than 3,3diphenylpropyl.

In the formula III compounds, where one of R 2

R

3 and R 4 is 6-fluoro, and the others are H, R 7 will be other than U.S. Application Serial No. 472,067, filed June 6, 1995 (file DC21e) discloses compounds of the structure R N N-

R

or 0 R1

R

2 1

R

4 or

R

6 y or Rs

I

Rs' 0 R6P O -0 I 9 WO 99/21564 WO 9921564PCTIUS98/21 750 or 0 0. 07 R,

NI

where 0is or -S- 11 0 X is: CHR 8 C- -CH- CH- or C- 0 R 9 RIO R 9

RIO

R

8

R

9 and R 10 are independently hydrogen, alkyl, alkenyl, alkynyl, aryl, azylalkyl, heteroaryl, heteroarylalkyl, cycloalkyl, or cycloalkylalkyl; Y "n Or -C 0 wherein 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 carbons, cycloalkyl, or cycloalkylalkyl wherein alkyl has at least 2 carbons, all optionally substituted through available carbon atoms with 1, 2, 3 or 4 groups selected from halo, haloalkyl, alkyl, alkenyl, alkoxy, aryloxy, aryl, arylalkyl, alkylmercapto, arylmercapto, cycloalkyl, cycloalkylalkyl, heteroary., fluorenyl, heteroarylalkyl, hydroxy or oxo; or R 1 is a fluorenyl-type group of the structure -R.11 -RIZ

-RI

z or z or- R 12 R 12 _-z 2

/\R

2 Net 13 RR4 A B C 10 WO 99/21564 WO 9921564PCT/US98/2 1750 16 Rs Het 1 or R-2 Het 2 D or

R

1 is an indenyl-type group of the structure

R

13 R1 31

R'

R--Z or R-Z'or R1- 2 1 5a '(CH 2 a R1aR 12 -t Z 2

R

6 a

E

(a 2,3 or 4) F

R

13 R13 ~R 14 R1 Het Het R11- Z or R12 1' 2 z 2 R 1 2 2 Risa (CH 2 )a G H RS

Z

1 and Z 2 are the same or different and are independently a bond, 0, S,

H

S S -NH-C- Or -Cwith the proviso that with respect to B, at least one of Z 1 and Z 2 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;

R

1 2 is hydrogen, alkyl, alkenyl, aryl, haloalkyl, trihaloalkyl, trihaloalkylalkyl, heteroaryl, heteroarylalkyl, arylalkyl, arylalkenyl, cycloalkyl, aryloxy, alkoxy, arylalkoxy or cycloalkylalkyl, with the provisos that 11 WO 9 9/21564 WO 9921564PCTIUS98/2 1750 when R 12 is H, aryloxy, alkoxy or arylalkoxy, -NH-C-

-C-

10 1 11 1 then Z 2 is 0 alkyl 0 0 or abond and when Z 2 is a bond, R 1 2 cannot be heteroaryl or heteroarylalkyl; Z is bond, 0, S, N-alkyl, N-aryl, or alkylene or alkenylene from 1 to 5 carbon atoms; R 1 3

R

1 4

R

1 5 and R 1 6 are independently hydrogen, alk-yl, halo, haloalkyl, aryl, cycloalkyl, cycloheteroalkyl, alkenyl, alkynyl, hydroxy, alkoxy, nitro, amino, thio, alkylsulfonyl, arylsulfonyl, alkylthio, arylthio, aminocarbonyl, alkylcarbonyloxy, arylcarbonylanino, alkylcarbonylamino, arylalkyl, heteroaryl, heteroarylalkyl or aryloxy; Rl 5 a and R 1 6 a are independently hydrogen, alkyl, halo, haloalkyl, aryl, cycloalkyl, cycloheteroalkyl, alkenyl, alkynyl, alkoxy, alkylsulfonyl, arylsulfonyl, alkylthio, arylthio, aminocarbonyl, alkylcarbonyloxy, arylcarbonylamino, alkylcarbonylanino, arylalkyl, heteroaryl, heteroarylalkyl, or aryloxy; or R 1 is a group of the structure

R

1 7 wherein p is 1 to 8 and R 17 and R 1 8 are each independently H, alkyl, alkenyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, cycloalkyl. or cycloalkylalkyl. at least one of R 1 7 and R 1 8 being other than H; or RI is a group of the structure

R

20 -R19 gK wherein R 19 is aryl or heteroaryl;

R

20 is aryl or heteroaryl;

R

21 is H, alkyl, aryl, alkylaryl, arylalkyl, aryloxy, arylalkoxy, heteroaryl, heteroarylalkyl, heteroarylalkoxy, cycloalkyl, cycloalkylalkyl or cyc loalkylalkoxy; 12 WO 99/21564 WO 9921564PCT/US98/21 750

R

2

R

3

R

4 are independently hydrogen, halo, alkyl, alkenyl alkoxy, aryloxy, aryl, arylalkyl, alkylmercapto, arylmercapto, cycloalkyl, cycloalkylalkyl, heteroaryl, heteroarylalkyl, hydroxy or haloalkyl;

R

5 is independently alkyl, alkenyl, alkynyl, aryl, alkoxy, aryloxy, arylalkoxy, heteroaryl, arylalkyl, heteroarylalkyl, cycloalkyl, cycloalkylalkyl, polycycloalkyl, polycycloalkylalkyl, cycloalkenyl, cycloheteroalkyl, heteroaryloxy, cycloalkenylalkyl, polycycloalkenyl, polycycloalkenylalkyl, heteroarylcarbonyl, amino, alkylanino, arylamino, heteroarylamino, cycloalkyloxy, cycloalkylamino, all optionally substituted through available 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, aminocarbonyl, alkynylaminocarbonyl, alkylaminocarbonyl, alkenylaminocarbonyl, alkylcarbonyloxy, arylcarbonyloxy, alkylcarbonylanino, arylcarbonylanino, arylsulfinyl, arylsulfinylalkyl, arylsulfonyl, alkylsulfonyl, arylsulfonylamino, heteroarylcarbonylamino, heteroarylsulfinyl, heteroarylthio, heteroarylsulfonyl, alkylsulfinyl;

R

6 is hydrogen or Cl-C 4 alkyl or Cl-C 4 alkenyl; all optionally substituted with 1, 2, 3 or 4 groups which may independently be any of the substituents listed in the definition of R 5 set out above; 13 WO 99/21564 PCT/US98/21750

R

7 is alkyl, aryl or arylalkyl wherein alkyl by itself or as part of arylalkyl is optionally substituted with oxo

J

(Het

V

Het and Ht are the same or different and are independently selected from heteroaryl containing 5- or 6-ring members; and 0 N-oxides 4u thereof; and pharmaceutically acceptable salts thereof; with the provisos that where in the first formula X is CH 2 and

R

2

R

3 and R 4 are each H, then R 1 will be other than 3,3diphenylpropyl, and in the fifth formula, where one of R 2

R

3 and R 4 is 6-fluoro, and the others are H, R 7 will be other than 4-(2-methoxyphenyl).

U.S. Provisional application 60/017,224 filed May 1996, discloses a compound which has the structure 0 R2'A) 1 B R (0)q

R

2 1a2- '1 B 11 I R 1 or

IA

on or

IB

including pharmaceutically acceptable salts thereof, Noxides thereof, wherein q is 0, 1 or 2; A is a bond; or

-N-

R

s where R 5 is H or lower alkyl, or R 5 together with R 2 forms a carbocyclic or heterocyclic ring system containing 4 to 8 members in the ring; B is a fluorenyl-type group of the structure 14 WO 99/21564 WO 9921564PCTIUS98/2 1750

R

3 R4

R

3 x

R

B is an indenyl-type group of the structure or

R

3

RT

R~

R

3 a (a 2,3 or 4)

R

3

R

Hiet 3 R 3a (CH 2 )a

R

3 Het

R

3 a RX is H, alkyl or aryl;

R

1 is alkyl, alkenyl, alkynyl, alkoxyl, (alkyl or aryl) 3 Si (where each alkyl or aryl group is independent), cycloalkyl, cycloalkenyl, substituted alkylamino, substituted arylalkylanino, aryl, arylalkyl, arylamino, aryloxy, heteroaryl, heteroarylanino, heteroaryloxy, arylsulfonylanino, heteroarylsulfonylanino, arylthio, arylsulfinyl, arylsulfonyl, alkylthio, alkylsulfinyl, alkylsulfonyl, heteroarylthio, heteroarylsulfinyl, heteroarylsulfonyl, -PO(R 1 3

(R

1 4 (where R 1 3 and R 1 4 are independently alkyl, aryl, alkoxy, aryloxy, heteroaryl, 15 WO 99/21564 PCT/US98/21750 heteroarylalkyl, heteroaryloxy, heteroarylalkoxy, cycloheteroalkyl, cycloheteroalkylalkyl, cycloheteroalkoxy, or cycloheteroalkylalkoxy); aminocarbonyl (where the amino may optionally be substituted with one or two aryl, alkyl or heteroaryl groups); cyano, 1,1-(alkoxy or aryloxy) 2alkyl (where the two aryl or alkyl substituents can be independently defined, or linked to one another to form a ring connected to L 1 (or L 2 in the case of R 2 at the 2position); 1,3-dioxane or 1,3-dioxolane connected to L 1 (or

L

2 in the case of R 2 at the 4-position; the R 1 group may optionally be substituted with 1, 2, 3 or 4 substituents, which can be any of the R 3 or R 1 groups or alkylcarbonylamino, cycloalkylcarbonylamino, arylcarbonylamino, heteroarylcarbonylamino, alkoxycarbonylamino, aryloxycarbonylamino, heteroaryloxylcarbonylamino, uriedo (where the uriedo nitrogens may optionally be substituted with alkyl, aryl or heteroaryl), heterocyclylcarbonylamino (where the heterocycle is connected to the carbonyl group via a nitrogen or carbon atom), alkylsulfonylamino, arylsulfonylamino, heteroarylsulfonylamino, 2 0

R

22 where J is: CHR 2 3 -CH-CE- or I I I I o R24 R 2 5

R

2 4

R

2

R

2 3

R

2 4 and R 2 5 are independently hydrogen, alkyl, alkenyl, alkynyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, cycloalkyl, or cycloalkylalkyl;

R

2 0

R

2 1

R

22 are independently hydrogen, halo, alkyl, alkenyl, alkoxy, aryloxy, aryl, arylalkyl, alkylmercapto, arylmercapto, cycloalkyl, cycloalkylalkyl, heteroaryl, heteroarylalkyl, hydroxy or haloalkyl; and 16 WO 99/21564 PCT/US98/21750 these substituents may either be directly attached to R 1 or attached via an alkylene at an open position;

R

2 is independently any of the groups set out for

R

1 H, polyhaloalkyl, or cycloheteroalkyl, and may be optionally substituted with one to four of any of the groups defined for R 3 or substituents defined for R 1

L

1 is a linking group containing from 1 to carbons in a linear chain including alkylene, alkenylene or alkynylene, which may contain, within the linking chain any of the following: one or two alkenes, one or two alkynes, an oxygen, an amino group, an oxo group, and may be substituted with one to five alkyl or halo groups;

L

2 may be the same or different from L 1 and may independently be any of the L 1 groups set out above or a singe bond;

R

3

R

3

R

4 and R 4 may be the same or different and are independently selected from H, halogen, CF 3 haloalkyl, hydroxy, alkoxy, alkyl, aryl, alkenyl, alkenyloxy, alkynyl, alkynyloxy, alkanoyl, nitro, amino, thiol, alkylthio, alkylsulfinyl, alkylsulfonyl, carboxy, alkoxycarbonyl, aminocarbonyl, alkylcarbonyloxy, alkylcarbonylamino, cycloheteroalkyl, cycloheteroalkylalkyl, cyano, Ar-, Aralkyl, ArO, Ar-amino, Ar-thio, Ar-sulfinyl, Ar-sulfonyl, Ar-carbonyl, Ar-carbonyloxy or Ar-carbonylamino, wherein Ar is aryl or heteroaryl and Ar may optionally include 1, 2 or 3 additional rings fused to Ar;

R

3a and R 3 b are the same or different and are independently any of the R 3 groups except hydroxy, nitro, amino or thio; and are the same or different and independently represent a or 6 membered heteroaryl ring which contains 1, 2, 3 or 4 heteroatoms in the ring which are independently N, S or 0; and including N-oxides; 17 WO 99/21564 PCT/US98/21750 X is a bond, or is one of the following groups: -s-

I

-0-

-N-

R

6 c

R

7

R

8

R

9 R 0 R9 -R1 c-C- I I

R

9

R

1 0 or

Y-

R

9

R

1 0 wherein Y is 0, N-R 6 or S; n' is 0, 1 or 2;

R

6 is H, lower alkyl, aryl, -C(0)-R 1 1 or -C -0-R 11

R

7 and R 8 are the same or different and are independently H, alkyl, aryl, halogen, -0-R 1 2 or

R

7 and R 8 together can be oxygen to form a ketone;

R

9

R

1 0

R

9 and R 1 0 are the same or different and are independently H, lower alkyl, aryl or -O-R 11

R

9 and R 10 are the same or different and are independently H, lower alkyl, aryl, halogen or

-O-R

11

R

11 is alky or aryl;

R

1 2 is H, alkyl or aryl; with the following provisos for compound of the structure 0 B R1 18 -19when R 1 is unsubstituted alkyl or unsubstituted arylalkyl, L 1 cannot contain amino; when R 1 is alkyl, L 1 cannot contain amino and oxo in adjacent positions (to form an amido group); when R 2

L

2 A- is H 2

R

1

L

1 cannot contain amino; when R 1 is cyano, L 1 must have more than 2 carbons.

R'L

1 must contain at least 3 carbons; with respect to compounds of formulas I, IA and IB, where R 1 is cycloheteroalkyl,

R

1 is exclusive of 1-piperidinyl, 1-pyrrolidinyl, 1-azetidinyl or 1-(2-oxo-pyrrolidinyl); with respect to the sulfur containing compounds and alcohols, R 2

L

2 cannot have an 0 or N atom directly attached to S=(O)q or CRX(OH), and for IA, R 2

L

2 cannot be H.

The discussion of the background to the invention herein is included to explain the context of the invention. This is not to be taken as an admission that any of the material referred to was published, known or part of the common general knowledge in Australia as at the priority date of any of the claims.

Throughout the description and claims of the specification the word "comprise" and variations of the word, such as "comprising" and "comprises", is not intended to exclude other additives, components, integers or steps.

Summary of the Invention In accordance with the present invention, novel compounds are provided which are inhibitors of NTP and have the structure 0 R.ABII L OR 0 o thereof wherein A is a bond or

-N-

R

where R 5 is H or lower alkyl or R 5 together with R 2 forms a carbocyclic or heterocyclic ring system containing 4 to 8 members in the ring.

W:\fiona\NKI\Marked Up\10876.doc WO 99/21564 WO 9921564PCT/US98/21 750 B is a fluorenyl-type group of the structure: (the above B is also referred to as a fluorenyl- type ring or moiety); or B is an indenyl-type group of the structure RR 3 R 3 a (CHa or R 3b

R

3

R

3 or Het R 3 (a =2,3 or 4) R 3a R 3 R 3 Het R b(the above B is also referred to as an indenyl-type ring or moiety);

R

3 a (CH2), Ra and Rb may be the same or different and can be hydrogen, alkyl, aryl, arylalkyl or heteroaryl (linked to the ring via a carbon atom); R~a is H, lower alkyl or aryl; RI is independently alkyl, alkenyl, alkynyl, aryl, alkoxy, aryloxy, arylalkoxy, heteroaryl, heteroazylalkoxy, arylalkyl, heteroarylalkyl, cycloalkyl, cycloalkylalkyl, polycycloalkyl, polycycloalkylalkyl, cycloalkenyl, cycloheteroalkyl, heteroaryloxy, cycloalkenylalkyl, polycycloalkenyl, polycycloalkenylalkyl, heteroarylcarbonyl, amino, alkylamino, arylamino, heteroarylamino, cycloalkyloxy, cycloalkylamino, all 20 WO 99/21564 WO 9921564PCT/US98/2 1750 optionally substituted through available 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, arylalk-ynyl, aryloxy, aryloxyalkyl, arylalkoxy, arylazo, heteroaryloxo, heteroarylalkyl, heteroarylalkenyl, heteroaryloxy, hydroxy, nitro, cyano, amino, substituted amino, thiol, alkylthio, arylthio, heteroarylthio, arylthioalkyl, alkylcarbonyl, arylcarbonyl, arylaminocarbonyl, alkoxycarbonyl, aininocarbonyl, alkynylaminocarbonyl, alkylaminocarbonyl, alkenyl-aminocarbonyl, alkylcarbonyloxy, arylcarbonyloxy, alkylcarbonylamino, arylcarbonylamino, arylsulfinyl, arylsulfinylalkyl, arylsulfonyl, alkylsulfonyl, arylsulfonylamino, heteroarylcarbonylamino, heteroarylsulfinyl, heteroarylthio, heteroarylsulfonyl, alkylsulfinyl; or RI and R 5 a can be joined to form a ring of the structure R0 0 where J is: CH23, c- -CHN-CH- or C-; I I I~ I I I 0 R 2 4

R

25

R

2 4

R

2

R

2 3

R

2 4 and R 25 are independently hydrogen, alkyl, alkenyl, alkynyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, cycloalkyl, or cycloalkylalkyl;

R

2 0

R

2 1

R

2 2 are independently hydrogen, halo, alkyl, alkenyl, alkoxy, aryloxy, aryl, arylalkyl, alkylmercapto, arylmercapto, cycloalkyl, cycloalkylalkyl, heteroaryl, heteroarylalkyl, hydroxy or haloalkyl; and these preferred substituents may either be directly 21 WO 99/21564 PCT/US98/21750 attached to R 1 or attached via an alkylene chain at an open position.

R

2 is hydrogen, alkyl, alkenyl, alkynyl, haloalkyl, alkoxyl, (alkyl or aryl) 3 Si (where each alkyl or aryl group is independent), cycloalkyl, cycloalkenyl, substituted alkylamino, substituted arylalkylamino, aryl, arylalkyl, arylamino, aryloxy, heteroaryl, heteroarylamino, heteroaryloxy, arylsulfonylamino, heteroarylsulfonylamino, arylthio, arylsulfinyl, arylsulfonyl, alkylthio, alkylsulfinyl, alkylsulfonyl, heteroarylthio, heteroarylsulfinyl, heteroarylsulfonyl, cycloheteroalkyl, cycloheteroalkylalkyl,

-PO(R

13

(R

14 (where R 1 3 and R 1 4 are independently alkyl, aryl, alkoxy, aryloxy, heteroaryl, heteroarylalkyl, heteroaryloxy, heteroarylalkoxy, cycloheteroalkyl, cycloheteroalkylalkyl, cycloheteroalkoxy, or cycloheteroalkoxyalkyl); or cyano, 1,1-(alkoxyl or aryloxy) 2 alkyl (where the two aryl or alkyl substituents can be independently defined.

The R 2 group may optionally have from one to four substituents, which include any of the R 1 substituents, and any of the preferred R 2 substituents set out below.

Preferred R 2 group when L 2 and A are each a single bond include the following: haloalkylamino, (where halo includes CF 3 alkylamino, cycloalkylamino, arylamino, heteroarylamino, alkoxyamino, aryloxyamino, heteroaryloxyamino, heterocyclylamino (where the heterocycle is connected to the carbonyl group via a nitrogen or carbon atom).

L

1 is a linking group containing from 1 to carbons in a linear chain (including alkylene, alkenylene or alkynylene), which may contain, within the linking chain any of the following: one or two alkenes, one or two alkynes, an oxygen, an amino group optionally substituted 22 WO 99/21564 PCT/US98/21750 with alkyl or aryl, an oxo group; and may be substituted with one to five alkyl or halo groups (preferably F).

L

2 may be the same or different from L 1 and may independently be any of the L 1 groups set out above or a single bond.

R

3

R

3

R

4 and R 4 may be the same or different and are independently selected from H, halogen, CF 3 haloalkyl, hydroxy, alkoxy, alkyl, aryl, alkenyl, alkenyloxy, alkynyl, alkynyloxy, alkanoyl, nitro, amino, thiol, alkylthio, alkylsulfinyl, alkylsulfonyl, carboxy, alkoxycarbonyl, aminocarbonyl, alkylcarbonyloxy, alkylcarbonylamino, cycloheteroalkyl, cycloheteroalkylalkyl, cyano, Ar, Aralkyl, ArO, Ar-amino, Ar-thio, Ar-sulfinyl, Ar-sulfonyl, Ar-carbonyl, Ar-carbonyloxy or Ar-carbonylamino, wherein Ar is aryl or heteroaryl and Ar may optionally include 1, 2 or 3 additional rings fused to Ar;

R

3 a and R3b are the same or different and are independently any of the R 3 groups; 4 and are the same or different and independently represent a or 6 membered heteroaryl ring which may contain 1, 2, 3 or 4 heteroatoms in the ring which are independently N, S or 0; and including N-oxides.

X (in the fluorenyl type ring) is a bond, or is one of the following groups: -o-

-N-

R

6 c-

R

7

R

8 23 WO 99/21564 PCT/US98/21750 C C

R

9 R1 0

R

9

R

10 C C- I I

R

9

R

10

Y--

R

9

R

10 wherein Y is 0, N-R 6 or S; n' is 0, 1 or 2;

R

6 is H, lower alkyl, aryl, -C(0)-R 1 1 or -C -O-R 11

R

7 and R 8 are the same or different and are independently H, alkyl, aryl, halogen, -0-R 12 or

R

7 and R 8 together can be oxygen to form a ketone;

R

9

R

1 0

R

9 and R 1 0 are the same or different and are independently H, lower alkyl, aryl or -O-R 11

R

9 and R 1 0 are the same or different and are independently H, lower alkyl, aryl, halogen or -0-R 1 1

R

1 1 is alky or aryl;

R

12 is H, alkyl or aryl; with the proviso that when A is a bond, R 2

L

2 cannot be hydrogen.

The pharmaceutically acceptable salts of the compounds of formula I include 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-butyl-amine, toctylamine, dehydroabietylamine, 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, and prodrug esters thereof.

24 WO 99/21564 PCT/US98/21750 In addition, in accordance with the present invention, a method for preventing, inhibiting or treating atherosclerosis, pancreatitis, hyperglycemia or obesity is provided, wherein a compound of formula I, IA or IB as defined hereinbefore is administered in an amount which decreases the activity of microsomal triglyceride transfer protein.

Furthermore, in accordance with the present invention, a method is provided for lowering serum lipid levels, cholesterol and/or triglycerides, or inhibiting and/or treating hyperlipemia, hyperlipidemia, hyperlipoproteinemia, hypercholesterolemia, atherosclerosis, hyperglycemia, pancreatitis, obesity, hypertriglyceridemia, Type II diabetes (NIDDM) wherein a compound of formula I as defined hereinbefore is administered in an amount which decreases the activity of microsomal triglyceride transfer protein.

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 if obtained from an organism cows, humans, etc.), can be isolated from the microsomal fraction of homogenized tissue; and stimulates the transport of triglycerides, cholesterol esters, or phospholipids from synthetic 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 127, 632-634 (1987)] which may have similar catalytic properties.

The phrase "stabilizing" atherosclerosis as used in the present application refers to slowing down the development of and/or inhibiting the formation of new atherosclerotic lesions.

25 WO 99/21564 PCT/US98/21750 The phrase "causing the regression of" atherosclerosis as used in the present application refers to reducing and/or 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 carbons, more preferably 1 to 12 carbons, in the normal chain,such as methyl, ethyl, propyl, isopropyl, butyl, tbutyl, isobutyl, pentyl, hexyl, isohexyl, heptyl, 4,4dimethylpentyl, octyl, 2,2,4-trimethylpentyl, nonyl, decyl, undecyl, dodecyl, the various branched chain isomers thereof, and the like as well as such groups including 1 to 4 substituents which may be any of the R 3 groups, or the R 1 substituents set out herein.

Unless otherwise indicated, the term "cycloalkyl" as employed herein alone or as part of another group includes saturated or partially unsaturated (containing 1 or 2 double bonds) cyclic hydrocarbon groups containing 1 to 3 rings, including monocyclicalkyl, bicyclicalkyl and tricyclicalkyl, containing a total of 3 to 20 carbons forming the rings, preferably 4 to 12 carbons, forming the ring and which may be fused to 1 aromatic ring as described for aryl, which include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclodecyl and cyclododecyl, cyclohexenyl, any of which groups may be optionally substituted with 1 to 4 substituents which may be any of the R 3 groups, or the R 1 substituents set out herein.

The term "cycloalkenyl" as employed herein alone or as part of another group refers to cyclic hydrocarbons containing 5 to 20 carbons, preferably 6 to 12 carbons and 26 WO 99/21564 PCT/US98/21750 1 or 2 double bonds. Exemplary cycloalkenyl groups include cyclopentenyl, cyclohexenyl, cycloheptenyl, cyclooctenyl, cyclohexadienyl, and cycloheptadienyl, which may be optionally substituted as defined for cycloalkyl.

The 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 to 3 bridges, preferably 6 to 12 carbons and 1 or 2 bridges.

Exemplary polycycloalkyl groups include bicyclooctanyl, adamantanyl, [2.2.1]-bicycloheptanyl, [2.2.2]-bicyclooctanyl and the like and may be optionally substituted as defined for cycloalkyl.

The term "polycycloalkenyl" as employed herein alone or as part of another group refers to a bridged multicyclic group containing 5 to 20 carbons and containing 0 to 3 bridges and containing 1 or 2 double bonds, preferably 6 to 12 carbons and 1 or 2 bridges. Exemplary polycycloalkyl groups include [3.3.0]-bicyclooctenyl, bicycloheptenyl, [2.2.2]-bicyclooctenyl and the like and may be optionally substituted as defined for cycloalkyl.

The term "aryl" as employed herein alone or as part of another group refers to monocyclic and bicyclic aromatic groups containing 6 to 10 carbons in the ring portion (such as phenyl or naphthyl) and may optionally include one to three additional rings fused to Ar (such as aryl, cycloalkyl, heteroaryl or cycloheteroalkyl rings) and may be optionally substituted through available carbon atoms with 1, 2, or 3 groups selected from hydrogen, halo, haloalkyl, alkyl, haloalkyl, alkoxy, haloalkoxy, alkenyl, trifluoromethyl, trifluoromethoxy, alkynyl, cycloalkylalkyl, cycloheteroalkyl, cycloheteroalkylalkyl, aryl, heteroaryl, arylalkyl, aryloxy, aryloxyalkyl, arylalkoxy, arylthio, arylazo, heteroarylalkyl, heteroarylalkenyl, heteroarylheteroaryl, heteroaryloxy, hydroxy, nitro, cyano, amino, substituted amino wherein the amino includes 1 or 2 substituents (which are alkyl, aryl or any of the other aryl compounds mentioned in the definitions), thiol, 27 WO 99/21564 PCT/US98/21750 alkylthio, arylthio, heteroarylthio, arylthioalkyl, alkoxyarylthio, alkylcarbonyl, arylcarbonyl, alkylaminocarbonyl, arylaminocarbonyl, alkoxycarbonyl, aminocarbonyl, alkylcarbonyloxy, arylcarbonyloxy, alkylcarbonylamino, arylcarbonylamino, arylsulfinyl, arylsulfinylalkyl, arylsulfonylamino or arylsulfonaminocarbonyl or any of the R 3 groups, or the R 1 substituents set out herein.

The 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 substituent, 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.

The term "amino" as employed herein alone or as part of another group may optionally be substituted with one or two substituents such as alkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, cycloheteroalkyl, cycloheteroalkylalkyl and/or cycloalkyl.

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", "alkylamino", "arylamino", or "arylalkylamino" as employed herein alone or as part of another group includes any of the above alkyl, aryl or arylalkyl groups linked to a nitrogen atom.

The term "acyl" as employed herein by itself or part of another group, as defined herein, refers to an organic radical linked to a carbonyl C group; examples of acyl groups include alkanoyl, alkenoyl, aroyl, aralkanoyl, heteroaroyl, cycloalkanoyl, and the like.

28 WO 99/21564 PCT/US98/21750 The term "alkanoyl" as used herein alone or as part of another group refers to alkyl linked to a carbonyl group.

Unless otherwise indicated, the term "lower alkenyl" or "alkenyl" as used herein by itself or as part of another group refers to straight or branched chain radicals of 2 to carbons, preferably 3 to 12 carbons, and more preferably 1 to 8 carbons in the normal chain, which include one to six double bonds in the normal chain, such as vinyl, 2propenyl, 3-butenyl, 2-butenyl, 4-pentenyl, 3-pentenyl, 2hexenyl, 3-hexenyl, 2-heptenyl, 3-heptenyl, 4-heptenyl, 3octenyl, 3-nonenyl, 4-decenyl, 3-undecenyl, 4-dodecenyl, 4,8,12-tetradecatrienyl, and the like, and which may be optionally substituted with 1 to 4 substituents, namely, halogen, haloalkyl, alkyl, alkoxy, alkenyl, alkynyl, aryl, arylalkyl, cycloalkyl, amino, hydroxy, heteroaryl, cycloheteroalkyl, alkanoylamino, alkylamido, arylcarbonylamino, nitro, cyano, thiol, alkylthio or any of the R 3 groups, or the R 1 substituents set out herein.

Unless otherwise indicated, the term "lower alkynyl" or "alkynyl" as used herein by itself or as part of another group refers to straight or branched chain radicals of 2 to carbons, preferably 2 to 12 carbons and more preferably 2 to 8 carbons in the normal chain, which include one triple bond in the normal chain, such as 2-propynyl, 3butynyl, 2-butynyl, 4-pentynyl, 3-pentynyl, 2-hexynyl, 3hexynyl, 2-heptynyl, 3-heptynyl, 4-heptynyl, 3-octynyl, 3nonynyl, 4-decynyl,3-undecynyl, 4-dodecynyl and the like, and which may be optionally substituted with 1 to 4 substituents, namely, halogen, haloalkyl, alkyl, alkoxy, alkenyl, alkynyl, aryl, arylalkyl, cycloalkyl, amino, heteroaryl, cycloheteroalkyl, hydroxy, alkanoylamino, alkylamido, arylcarbonylamino, nitro, cyano, thiol, and/or alkylthio, or any of the R 3 groups, or the R 1 substituents set out herein.

The term "alkylene" as employed herein alone or as part of another group refers to alkyl groups as defined 29 WO 99/21564 PCTIUS98/21 750 above having single bonds for attachment to other groups at two different carbon atoms and may optionally be substituted as defined above for "alkyl".

The terms "alkenylene", and "alkynylene" as employed herein alone or as part Of another group refer to alkenyl groups as defined above and alkynyl groups as defined above, respectively, having single bonds for attachment at two different carbon atoms.

Suitable alkylene, alkenylene or alkynylene groups or (CH2)m, (CH2)n or (CH 2 )p (which may include alkylene, alkenylene or alkynylene groups) as defined herein, may optionally include 1, 2, or 3 substituents which include any of the R 3 groups, or the RI substituents set out herein.

Examples of alkylene, alkenylene and alkynylene include

-CXECH-.CH

2 9 CE 2 9 -C C-CE 2 9 -C3 2

C-

I I 0

-CH

2

-CH

2

CH

2

C-

I I 0

CH

3 -C=CH CE 2

CH

2 C CCH 2

(CE

2 2 9 -(CH 2 3 9 -(CE 2 4

CH

3 M3(C 2 2 CCH- -CE 2

CE-

C

3 3

C

2 CHC3 2

CAH

CH

3

-CHERCH

2

E-

CI'

-CHCHCH

2

K

3

C

3 3

CE

3

-CE

2

-C-CE

2

CE

3

F

(C3 2 5 -(CE3 2 2

-C-CE

2

F

30 WO 99/21564 PCT/US98/21750 C1 CH3 CE3

CH

2

-CH-C

2

(CH

2 2 -CH- 9 -CH 2 9 I

I

CHa 3 C 3

C

2 C -CH -CH-C H 2 I I I I CH3 CH3 C8 3 CH3 CH3

OCH

3

I

-cH"CH2CH 2 CH-CH2CH 2 ECH 2 CH2-

OCH

2 CE2- CHEHC-

NHC

2

CE

2

CH

3 CE 2

CH

2 I

I

(CH

2 3

-CF

2

-CH

2

-N-CH

2 or CE 3 The term "halogen" or "halo" as used herein alone or as part of another group refers to chlorine, bromine, fluorine, and iodine as well as CF3, with chlorine or fluorine being preferred.

The term "metal ion" refers to alkali metal ions such as sodium, potassium or lithium and alkaline earth metal ions such as magnesium and calcium, as well as zinc and aluminum.

The term "heterocyclyl" as used herein refers to "cycloheteroalkyl" groups and "heteroaryl" groups as defined herein.

The term "cycloheteroalkyl" as used herein alone or as part of another group refers to a 6- or 7-membered saturated or partially unsaturated ring which includes 1 to 2 hetero atoms such as nitrogen, oxygen and/or sulfur, linked through a carbon atom or a heteroatom, where possible, optionally via the linker (CH2)p (which is defined above), such as 31 WO 99/21564 PCT/US98/21750

S'

0 N 0 and the like. The above groups may include 1 to 4 substituents such as alkyl, halo, oxo and/or any of of the

R

3 groups, or the R 1 substituents set out herein. In addition, any of the above rings can be fused to a cycloalkyl, aryl, heteroaryl or cycloheteroalkyl ring.

The term "cycloheteroalkoxy" as used herein alone or as part of another group refers to a 6- or 7membered saturated or partially saturated ring which includes at least one oxygen atom in the ring and at least 1 or 2 other hetero atoms in the ring such as nitrogen, oxygen and/or sulfur, linked through a carbon or heteroatom, where possible, optionally via the linker (CH2)p, such as 0 and the like. The above groups may include 1 to 4 substituents such as alkyl, halo, oxo and/or any of of the

R

3 groups, or the R 1 substituents set out herein. In 32 WO 99/21564 PCT/US98/21750 addition, any of the above rings can be fused to a cycloalkyl, aryl, heteroaryl or cycloheteroalkyl ring.

The term "heteroaryl" as used herein alone or as part of another group refers to a 5- or 6- membered aromatic ring which includes 1, 2, 3 or 4 hetero atoms such as nitrogen, oxygen or sulfur,and such rings fused to an aryl, cycloalkyl, heteroaryl or cycloheteroalkyl ring (e.g.

benzothiophenyl, indolyl), and includes possible N-oxides, such as 0 0CJ) 0 NS N L I (TV\ H r S N N N NN 'N 9 N N H

N

N-N N-N

N-N

S 9 1 I ON and the like.

Ar may be either aryl or heteroaryl as defined above.

J

J

>4 and are the same or different, as defined hereinbefore, and are attached to the central ring of the indenyl or fluorenyl type group at adjacent positions (that is, ortho or 1,2positions). Examples of such groups include 33 WO 99/21564 PCT/US98/21750

R

7a is H, lower alkyl, aryl, -C R 7b -C OR 7b 0

R

7 b is alkyl or aryl optionally include 1 to 4 substituents such as any of the NUJ u^ N wherein u is selected from 0, S cand 7 ;be fused to a

R

7 c is H, lower alkyl, aryl, -Ceteroaryl oR 7 cycloh, -CeterOR 7 ;ing.

R

7 b is alkyl or aryl.

The heterm cycloheteroalkylalkyups including the above groups may optionally include 1 to 4 substituents such as any of the

R

3 groups, or the R 1 substituents set out herein. In addition, any of the above rings can be fused to a cycloalkyl, aryl, heteroaryl or cycloheteroalkyl ring.

The term cycloheteroalkylalkyl" as used herein alone or as part of another group refers to cycloheteroalkyl groups as defined above linked through a C atom or heteroatom to a (CH 2 )p chain.

The term "heteroarylalkyl" or "heteroaryl-alkenyl" as used herein alone or as part of another group refers to 34 WO 99/21564 PCT/US98/21750 a heteroaryl group as defined above linked through a C atom or heteroatom to a -(CH 2 p chain, alkylene or alkenylene as defined above.

The term "polyhaloalkyl" as used herein refers to an "alkyl" group as defined above which includes from 2 to 9, preferably from 2 to 5, halo substituents, such as F or Cl, preferably F, such as CF 3

CH

2

CF

3 or CF 3

CF

2

CH

2 Preferred are compounds of formula I wherein A is

NH,

B is

R

3 R4 X is a bond, oxygen or sulfur; R 3 and R 4 are independently H or F.

Preferred R 2 groups are alkyl, polyfluoroalkyl (such as 1,1,1-trifluoroethyl), alkenyl, aryl, heteroaryl or heteroarylalkyl, such as 2-pyridylmethyl (preferably substituted with one of the preferred R 2 substituents above).

It is preferred that L 1 contains 2 to 7 atoms in the linear chain and L 2 is a bond or lower alkylene.

The preferred L 2 group is alkylene such as CH 2 or a bond.

The preferred A group is -NH-.

Preferred R 1 groups are as set out in the preferred compounds shown in the following Table.

More preferred are compounds of the invention which have the following structures wherein R 1 CO, n and the geometry are as indicated below.

35 wo 99/21564 WO 9921564PCT/UJS98/21 750 Preferred Structures of Cyclic Phosphonates n geometry R 1 CO n geometry R'CO

CF

3

CO

CF

3 Co

QAO

trans 1 cis 1 trans 2 trans C6-H a2yCO 1 trans 1 trans 1 trans

SN

CO

0

CO

trans 1 trans 2 cis 0 11

OC-

trans 36 WO 99/21564 PCTIUS98/21750 The compounds of formula I may be prepared by the exemplary processes described in the following reaction schemes. Exemplary reagents and procedures for these reactions appear hereinafter and in the working Examples.

Methods for preparing starting materials useful in preparing cyclophosphonates in accordance with the present invention, are shown in the folowing reaction schemes.

Scheme 1.

OH

Acylation (carbodiimide coupling)

RICO

2 H, RIN=C=NR 2 ROBt Ra

OR

NMCOR

1

OH

3 Ra O 18. y-s Rb

>O

O2 2 Acylation RaOCORI Ra O 3 R'COC1, 3 Et3N Solvolysis 3RCOCI, 3 Et 3 N NCOR 1 MeOH, NaB NCOR 1 Rb OCOR b O Silylation Ra OSi(CH 3 3

[(CH

3 3 Si] 2 1N TXSBz

U

Rb Osi(CH 3 3 1. Acylation Ra OH (RCOC1, Et 3

N)

2. Methanolysis) NCOR1 (MeOH, HC1) sa Rb OH As seen in Scheme 1A, serinol derivative can be acylated via carbodiimide coupling directly to 3.

Alternatively, as seen in Scheme 1B, 2 can be acylated with acid chlorides to tri-acylated 4. Subsequent solvolysis provides diols 3. As seen in Scheme 1C, the diols could also be prepared from bis-0-trimethylsilylated amine acylation with acid chlorides and subsequent methanolysis to provide 3.

It will be appreciated that in the reactions to follow, unless otherwise indicated, the moiety in the starting materials, intermediates and final products is set out as 37 WO 99/21564 PCT/US98/21750 for purposes of illustration only.

It will be appreciated that the moiety in the starting materials, intermediates and final products in all reactions set forth herein, unless indicated to the contrary, may be any of the fluorenyl-type groups as well as any of indenyl-type groups

R

3 R3a (CH a (a 2,3 or 4) or R3R

SR

3 b Raa

R

3 R3 R3' H R3' Htet Het f f or R 3b R3b

R

3 a (CH 2 )a R3a The above B moieties (including all fluorenyl-type groups and all indenyl-type groups) are collectively referred to as "fluorenyl-type" moieties. The use of the first fluorenyl-type group (as set out in the previous 38 WO 99/21564 PCT/US98/21750 paragraph) in the Reaction Schemes is for purposes of illustration only; any of the 3 fluorenyl groups or 4 indenyl groups as set out above may be employed in any of the Reaction Schemes set out herein in place of The compounds of formula I of the invention may be prepared as shown in Reaction Scheme 2.

Scheme 2 1. TMSBr 2. C1COCOC1 O L ",0alkyl i-NCOR', amine base A Oalkyl Rb O (3

L,

2

R

2

I

K- I o/ -II Ra O L 1

-P

0 Ll-_ P

O-

r 0 A O

L

2

R

2 R I H RSa H R 5 s 7-cis io 2-trans zp As seen in Scheme 2, dialkyl phosphonate ester Im is treated with bromotrimethylsilane and then is treated with oxalyl chloride and dimethylformamide (employing conventional procedures) to give a diphosphinyl dichloride intermediate which (without purification) is reacted with diol 3 (which may be chiral or achiral) (as formed in Scheme lA, 1B or IC) and an amine base such as triethylamine to give a mixture of the 7-trans and 7-cis isomers (lo and Ip, respectively) of the invention. The 7trans and 7-cis isomers may be separated by conventional chromatographic techniques.

39 WO 99/21564 PCT/US98/21 750 Scheme 3 Preparation of other Acyl Derivatives From 7 -trans Compounds -~0 '0 AR

NRI.COR.

L

2

R

2 Ib H RS& (where Rl' is an arylmethyloxy or (7 -trans) beteroarylmethyloxy) Hyrdrogenolysis 1. H 2 Pd-C/EtOH 2. R1CO 2 H, R 1

.N=C=NR

2 HRO~t I I -0 0 Ab1

NCOR

1 1 2

R

2 Rh I H R 5 7-trans As seen in Scheme 3, the 7-trans isomer (Rl 1

=OCH

2

C

6

H

5 (and the corresponding 7-cis isomer) may be made to undergo hydrogenolysis to form an amine intermediate which can be reacted with R'CO 2 H to prepare other acyl derivatives in accordance with the invention.

The starting material Im may be prepared according to the following Reaction Schemes 4 and 4A.

40 WO 99/21564 WO 9921564PCT/US98/21 750 Reaction Schemne 4 1) base 2) Ha-L.

1

-Y

Y=Hal or 0-PG

A

1 2

A

V711I

L

2

R

lh (Y=Hal) XIVa (Y=OPG)

Y=OPG

Deprotection F- (for example, n-BU 4

NF)

A

1 2 2 3g Halide Formation/

A

L

2

R

2

I

where PG is an oxygen protecting group, as t-Bu(CH 3 2 Si Or tBu(Ph) 2 Si- Arbuzov 05 Reaction Q% such Oalky Qalkyl Excess (AlkylO) 2

P(OQ')

0 C to 1 80 0

C

A

I TM

L

2

R

2 (01 is alkyl, triorganosilyl (such as trimethylsilyl or t-butyldimethylsily), H, the latter in the presence of base such as butyllithium, sodium hydride, or sodium bis-(trimethylsilylamide) 41 WO 99/21564 PCT/US98/21750 Reaction Scheme 4A (Alternative Method for Making Ih) 1) base 2) HaI-L 1 -Y I 1 3)CICOCOCII DMF N-Rs 0 OH 4) (R 2

L

2

)R

5 NH L 2

R

2 II Ih' Protected alcohol XIVa can be converted into a wide variety of functional groups through the intermediacy of a halide Ih. For example, the alcohol Ig can be converted to the halide Ih of the invention by either activation through the sulfonate ester (tosyl chloride, or mesyl chloride) and iodide displacement (Nal or KI in acetone or 2-butanone), or by reaction with triphenylphosphine, 12 and imidazole.

The halide Ih can undergo an Arbuzov reaction to form phosphonates, phosphinates and phosphine oxides of the invention Im. The Arbuzov reaction can be accomplished with phosphites, phosphinites, and phosphonites (for example, (alkylO) 3 P or (alkylO)2POSi(alkyl) 3 or (alkylO) 2 POH, the latter being in the presence of a base such as butyllithium, sodium hydride or sodium bis(trimethylsilylamide)) at temperatures within the range from about -20 0 C to about 180 0

C.

The starting material VIIIA for compound Im may be prepared as shown in Reaction Scheme 42 WO 99/21564 PCT/US98/21750 Reaction Scheme 5 (Amides) Preparation of Compounds of Formula I where A is

-N-

R

0 COOH R L C 1) amide formation s e.g. 1) acid chloride formation X 2) (R 2

L

2 )RSNH

X

II VII1A A= -N-

R

s As seen in Scheme 5, the amide VIIIA may be formed by treating II with thionyl chloride or oxalyl chloride in an inert organic solvent such as dichloromethane (optionally in the presence of dimethylformamide (DMF)) to form the acid chloride IIA 0

II

11A Acid chloride IIA, without separation from the reaction mixture, is treated with amine (R 2

L

2

)R

5 NH at a reduced temperature within the range from about -40 0 C to about room temperature, to form the amide VIIIA.

Amide VIIIA may be converted to compounds of formula I employing the procedure set out in Reaction Scheme 4/4A.

In carrying out the above reaction to form amide VIIIA, the amine will be employed in a molar ratio to acid chloride IIA within the range from about 4:1, to about 1:1, optionally in the presence of a tertiary amine base or other acid scavenger.

Alternative formation of amide VIIIA from acid II and (R 2

L

2

)R

5 NH can be carried out via standard literature procedures.

43 WO 99/21564 PCT/US98/21750 Reaction Scheme 6 (Class Esters) Preparation of Esters VIIIA (A 0

HO

2 H 1) Acid, R 2

L

2 OH or R 2 L--C H 2) (COCI) 2

R

2

L

2

OH

or 3) DCC, HOBT, X DMAP, R 2

L

2

OH

esterification As seen in Reaction Scheme 6, ester compounds of formula VIIIB wherein A oxygen can be prepared by an acid catalyzed esterification of acid II employing an acid such as H 2

SO

4 or p-toluene-sulfonic acid in the presence of an alcohol such as allyl alcohol, ethanol or methanol.

Alternatively, activation of the acid II to the acid chloride (with oxaly chloride or thionyl chloride) followed by treatment with an alcohol optionally in the presence of a tertiary amine base or other acid scavenger, gives compounds of formula VIIIB.

Various additional methods of activation include mixed anhydride formation ((CF 3

COO)

2 or i-BuOCOC1) or formation of the acylimidazole (carbonyldiimidazole) or with DCC and HOBT in the presence of DMAP (4-dimethylaminopyridine). These activated intermediates readily form esters upon treatment with alcohols.

44 WO 99/21564 PCT/US98/21750 Reaction Scheme 7 (Amides from Isocvanates) Preparation of Amides VIIIC (A is NH) C7 1) base 2) R 2

L

2

NCO

0 R2L2HN" VizzC Compounds of formula VIIIC where A is -NH- (amides) can be prepared by the methods shown in Reaction Scheme 7 from known compound IV.

Treatment of compound IV with base, such as n-BuLi, followed by reacting the anion with an isocyanate gives compound VIIIC.

Scheme 8 Scheme for rcomrnouni Tm9 Alternate Scheme for Comnound Im2 Deesterification TMSBr or TMSI (Optional tertiary amine base) oo O==1

OTMS

R2L/A

OTMS

R

2 2 Disilyl Ester Intermediate Phosphonate Ester Formation

(COCI)

2 DMF, CH 2

CI

2

-M-

lo or Ip Compounds Im may be modified by the various transformations set out in Reaction Scheme 8.

The compounds of the invention may be employed in preventing, stabilizing or causing regression of atherosclerosis in a mammalian species by administering a therapeutically effective amount of a compound to decrease the activity of MTP.

45 WO 99/21564 PCTUS98/21750 The compounds of the invention can be tested for MTP inhibitory activity employing the procedures set out in U.S. application Serial No. 117,362 filed September 3, 1993, employing MTP isolated from one of the following sources: bovine liver microsomes, HepG 2 cells (human hepatoma cells) or recombinant human MTP expressed in baculovirus.

The compounds of the invention may also be employed in lowering serum lipid levels, such as cholesterol or triglyceride (TG) levels, in a mammalian species, by administering a therapeutically effective amount of a compound to decrease the activity of MTP.

The compounds of the invention may be employed in the treatment of various other conditions or diseases using agents which decrease activity of MTP. For example, compounds of the invention decrease the amount or activity of MTP and therefore decrease serum cholesterol and TG levels, and TG, fatty acid and cholesterol absorption and thus are useful in treating hypercholesterolemia, hypertriglyceridemia, hyperlipemia, hyperlipoproteinemia, hyperlipidemia, pancreatitis, hyperglycemia, atherosclerosis, non-insulin dependent diabetes (Type II diabetes) and obesity.

The compounds of the present invention are agents that decrease the activity of MTP and can be administered to various mammalian species, such as monkeys, dogs, cats, rats, humans, etc., in need of such treatment. These agents can be administered systemically, such as orally or parenterally.

The agents that decrease the activity or amount of MTP can be incorporated in a conventional systemic dosage form, such as a tablet, capsule, elixir or injectable formulation. The above dosage forms will also include the necessary physiologically acceptable carrier material, excipient, lubricant, buffer, antibacterial, bulking agent 46 WO 99/21564 PCT/US98/21750 (such as mannitol), anti-oxidants (ascorbic acid or sodium bisulfite) or the like. Oral dosage forms are preferred, although parenteral forms are quite satisfactory as well.

The dose administered must be carefully adjusted according to the age, weight, and condition of the patient, as well as the route of administration, dosage form and regimen, and the desired result. In general, the dosage forms described above may be administered in amounts of from about 5 to about 500 mg per day in single or divided doses of one to four times daily.

The following Examples represent preferred embodiments of the invention. All temperatures are in °C unless indicated otherwise.

NOTE: The phrase "flash chromatography" refers to chromatography performed on EM Industries Silica Gel (catalog #9385-9), 230-400 mesh under 10-20 psi of nitrogen pressure.

Example 1 (E)-9-[4-[2-Oxo-5-[[[4'-(trifluoromethyl)[1,1 '-biphenyl]-2-yl]carbonyl]amino]- 1,3,2-dioxaphosphorinan-2-yl]butyl]-N-(2,2,2-trifluoroethyl)-9H-fluorene- 9-carboxamide, N-oxide.

o 9 CF.

trans isomer 47 WO 99/21564 PCT/US98/21 750 Example 2 (Z)-9-[4-[2-Oxo-5-[[[4'-(trifluoromethyl)[1 ,1 -biphenyl]-2-yI]carbonyl]amino]- 1 3 2 -dioxaphosphonnan-2-yI]butyII-N-(2,2,2-trifluoroethyl)9H-fluorene- 9-carboxamide, N-oxide.

00

CICF

3

H

A.

OH OH

H

0

CF

3 To a stirred solution of 1.33 g (5.00 mmol) of 41trifluorophenyl-2-benzoic acid (Aldrich Chemical Co.), 0.455 g (5.00 mmol) of serinol (Aldrich Chemical Co.), 0.750 g (5.0 mnmol) of HOBt and 0.5 rnL (3.6 Inmol) of triethylainine in 10 InL of dichloromethane at room temperature under argon, was added 1.0 g (5.25 mmol) of EDCI, portionwise, over 3 min. After 16 h, the reaction mixture was diluted with ethyl acetate, washed once with saturated sodium bicarbonate solution, once with brine and once with 10% citric acid solution, dried (MgSO 4 and evaporated. Purification by flash chromatography on silica gel (5 x 15 cm column, ethyl acetate) provided title compound as a white solid, mp 146-148*C, 1.23 g, 72% yield.

48 WO 99/21564 PCT/US98/21750

B.

COt^CF 3 S/ V PO 3 Et 2 B(1).

0

OH

Br To a solution of 9-fluorenecarboxylic acid (Aldrich) g, 240 mmol) in THF (1200 mL) at 0 0 C was added dropwise a solution of n-butyllithium (2.5M, 211 mL, 530 mmol) in THF. The yellow reaction was stirred at 0 0 C for 1 h, then 1,4-dibromobutane (31.3 mL, 260 mmol) was added dropwise over 30 min. The reaction was stirred at 0oC for 30 min, then the reaction was warmed to RT for 30 h. The reaction was extracted with water (3 x 750 mL). The combined aqueous layers were extracted with ethyl ether (800 mL).

The aqueous layer was made acidic with HC1 solution (IN, 500 mL), then extracted with dichloromethane (3 x 750 mL).

The combined organic layers were dried over MgSO4.

Evaporation gave title compound (71 g, 85%) as a white solid.

B(2).

H

CF

3 Br To a solution of Part B(l) acid (60 g, 173 mmol) and DMF (100 pL) in CH 2 C12 (600 mL) under argon at 0OC was 49 WO 99/21564 PCT/US98/21750 added oxalyl chloride (104 mL, 2.OM in CH 2 C12, 208 mmol) dropwise. The reaction was stirred at O'C for 10 min, then warmed to RT and stirred for 1.5 h. The reaction was concentrated in vacuo to give the crude acid chloride as a yellow oil. To a suspension of 2 ,2,2-trifluoroethylamine hydrochloride (25.9 g, 191 mmol) in CH 2 C1 2 (500 mL) at 0 0

C

under argon was added triethylamine (73 mL, 521 mmol) followed by dropwise addition of a solution of the crude acid chloride in CH 2 C1 2 (15 mL). The reaction was stirred at O'C for 1 h, diluted with CH 2 C1 2 (500 mL), and washed with water (2 x 300 mL), IN HC1 (2 x 300 mL), saturated NaHC0 3 (2 x 300 mL), and brine (2 x 300 mL), then dried over MgSO 4 Evaporation gave 80 g of a oil which was purified by flash chromatography on silica gel (2.5 kg).

The crude product was loaded in a mixture of CH 2 C1 2 and hexane, and eluted with a step gradient of 10% EtOAc/hexane (4L) to 15% EtOAc/hexane (2L) to 20% EtOAc/hexane (4L).

Pure fractions were combined and evaporated to give title compound (52.5 g, 71%) as a white solid (mp 88-92'C).

B(3).

cot 4 c 2 Cv 3

PO

3 Et 2 A solution of 2.13 g (5.00 mmol) of Part B(2) compound in 3.5 mL of triethylphosphite under argon was heated to 110°C for 16 h and then to 180°C for 4 h. The reaction was cooled and then the volatiles were distilled off at 100°C at 1 Torr. The residue was purified by flash chromatography (5x15 cm column, EtOAc) to give title compound (1.92 g, 79%) as a waxy yellow solid (mp 87-89°C).

mp: 87-89°C MS (FAB) m/e 484 50 WO 99/21564 PCT/US98/21750 Anal. Cald'd for C24H 2 9

NO

4

PF

3 +0.13 mol H 2 0: C, 59.33; H, 6.07; N, 2.88; P, 6.37; F, 11.73 Found: C, 59.09; H, 5.98; N, 2.95; P, 6.51; F, 11.92.

C. 9 4 2 -Oxo-5-[[[4'-(trifluoromethyl)- [1,1'-biphenyl]-2-yl]carbonyl]amino]-1,3,2dioxaphosphorinan-2-yl]butyl]-N-(2,2,2trifluoroethyl)-9H-fluorene-9-carboxamide, N-oxide (Example 1) D. 9 4 2 -Oxo-5-[[[4'-(trifluoromethyl)- [1,1'-biphenyl]-2-yl]carbonyl]amino]-1,3,2dioxaphosphorinan-2-yl]butyl]-N-(2,2,2trifluoroethyl)- 9 H-fluorene-9-carboxamide, N-oxide (Example 2) To a stirred solution of 500 mg (1.03 mmol) of Part B compound in 10 mL of dichloromethane at room temperature under argon was added 0.45 mL (3.4 mmol) of bromotrimethylsilane. After 1 h, the reaction mixture was evaporated to provide a thick oil. The oil was dissolved in 5 mL of dichloromethane at room temperature under argon and treated with 0.26 mL (3.0 mmol) of oxalyl chloride and 100 lL of DMF. After 2 h, the reaction mixture was evaporated and redissolved in 10 mL of dichloromethane. To this solution at room temperature under argon, was added 340 mg (1.0 mmol) of Part A compound and 0.5 mL (3.6 mmol) of triethylamine. After 14 h, the reaction was diluted with ethyl acetate and washed once with 10% citric acid solution, dried (MgSOJ) and evaporated. Purification by flash chromatography on silica gel (5 x 15 cm column, 1 L 85:15 ethyl acetate/hexanes, then ethyl acetate) provided two fractions.

The less polar fraction was designated Part C (Example 1) compound, white solid, mp 105-107 0 C, 205 mg, 27% yield.

51 WO 99/21564 WO 9921564PCTfUS98/21 750 MICROANALYSIS Calculated for C 3 7

H

3 3

F

6

N

2 0 5 P 0.55 H 2 0: C, 60.01; H, 4.64; N, 3.78; F, 15.39; P, 4.18 Found: C, 60.00; H, 4.48; N, 3.64; F, 15.10; P, 3.75.

MS (electrospray, ions) m/e 731.

The more polar fraction was designated Part D (Example 2) compound, white solid, mp 102-104'C, 130 mg, 17%.

1 H NMR, 1 3 C NMR, IR and mass spectrometry were consistent for the indicated compound.

MICROANALYSIS Calculated for C 3 7

H

33

F

6

N

2 0 5 P 064 H 2 0: C, 59.88; H, 4.66; N, 3.77; F, 15.36; P, 4.17 Found: C, 59.88; H, 4.44; N, 3.75; F, 15.08; P, 4.14.

MS (electrospray, ions) m/e 731.

Examrle 3 trans-9-[4-[5-(Benzoytamino)-2-oxo-1 ,3,2-dioxaphosphorinan-2-y]butyl]- N-(2,2,2-trif luoroethyl)-9H-fluorene-9-carboxamide.

0- 0

HH

trans isomer A. N- 3 -Dibenzoxyloxy-2-propyl)benzamjde oHN-(OCOC Hs To a stirred slurry of 1.82 g (20.0 mmol) of serinol and 9.2 niL (66 mmol) of triethylamine in 20 niL of dichioromethane at -5*C under argon, was added dropwise, niL (61 nimol) of benzoyl chloride (Aldrich) over 30 min.

52 WO 99/21564 PCT/US98/21750 After 2 h, the reaction was diluted with ethyl acetate and washed three times with 1 M hydrochloric acid, once with brine, twice with saturated sodium bicarbonate solution, dried (MgSOj) and evaporated. The resulting white solid was recrystallized from ethyl acetate/hexane to provide title compound, mp 96-98 0 C, 5.62 g, 70% yield.

B. N-(1,3-Dihvdroxy-2-propyl) benzamide

OH

0o OH To a stirred solution of 160 mg (4.0 mmol) of sodium hydride in 50 mL of methanol at room temperature under argon, was added 4.03 g (10.0 mmol) of Part A compound. The slurry was heated to reflux where a clear solution forms. After 3 h, the reaction was cooled and mL of 4 N hydrogen chloride in dioxane was added.

Evaporation and re-evaporation from methanol onto silica gel (5 g) and purification by flash chromatography on silica gel (5 x 15 cm column, 3:47 methanol/ethyl acetate) gave title compound as white solid, 1.22 g, 62% yield.

C. trans-9-[4-[5-(Benzoylamino)-2-oxo-l,3,2dioxaphosphorinan-2-yl]butyl]-N-(2,2,2trifluoroethyl)-9H-fluorene-9-carboxamide To a stirred solution of 699 mg (1.45 mmol) of Example 1 Part B compound in 5 mL of dichloromethane at room temperature under argon was added 0.63 mL (4.8 mmol) of bromotrimethylsilane. After 1 h, the reaction mixture was evaporated to provide a thick oil. The oil was dissolved in 3 mL of dichloromethane at room temperature under argon and treated with 0.26 mL (3.0 mmol) of oxalyl chloride and 50 gL of DMF. After 2 h, the reaction mixture was evaporated and redissolved in 3 mL of dichloromethane.

53 WO 99/21564 WO 9921564PCT[US98/21 750 To this solution at room temperature under argon, was added 283 mg (1.45 nimol) of Example 3 Part B compound and 0.6 mL- (4.3 mmol) of triethylamine. After 2 h, the reaction was diluted with ethyl acetate and washed once with 10% citric acid solution, dried (MgSO 4 and evaporated. Purification by flash chromatography on silica gel (5 x 15 cm column, ethyl acetate) provided title compound, white solid, mp 166-168 0 C, 245 mg, 29% yield.

MICROANALYSIS Calculated for C 3 0

H

3

OF

3

N

2 0 5 P 0.5 H 2 0: C, 60.50; H, 5.25; N, 4.70; F, 9.57 Found: C, 60.47; H, 5.01; N, 4.62; F, 9.30.

MS (electrospray, ions) m/e 587.

Exaile 4 trans-9-[5-[2-Oxo-5-[14-(tifluoromethyl)[1 ,1 -biphenyl]-2-yI]carbonyl]amino]- 1 .3,2-dioxaphosphorinan-2-yI]pentyl]-N-(2,2,2-trfluoroethyl)..9H-fluorene-9 carboxamide.

0 3

CCF

3

N

HH

H

trans isomer Examiple cis-9-[5-[2-Oxo-5-[4'-(tifluoromethyl)[1 ,1 -biphenyll-2-yI]carbonyl]aminol- 1 ,3,2-dioxaphosphoinan-2-ylpentyu]-N-(2,2,2-trifluoroethyl)-9Hfluorene..

9-carboxamide.

0 N -C3CF 3 H A cis isomer 54 WO 99/21564 PCT/US98/21750 A. 2-Amino-I,3-bis (trimethylsilyloxy) propane OSi(CHAh

NH

2 OSi(CHAb A stirred solution of 1.84 g (20.0 mmol) of serinol in 4.25 mL (20.1 mmol) of hexamethyldisilazane containing 1 drop of bromotrimethylsilane was heated to 180°C under argon. After 16 h, the reaction mixture was partially cooled and distilled at 0.6 Torr, collecting the fraction with bp 44-47°C to give title compound as a colorless liquid, 4.05 g, 86%.

B. N-[(l,3-Dihydroxy)-2-propyl]-2-(4-trifluoromethylphenyl)benzamide OH OH 0

CF

3 A solution of 4'-trifluoromethyl-2-biphenylcarbonyl chloride [prepared from 4.40 g (16.5 mmol) of the corresponding carboxylic acid] in 10 mL of THF was added dropwise to a solution of 3.88 g (16.5 mmol) of Part A compound and 2.40 mL (17.2 mmol) of triethylamine in 20 mL of THF at 0C over 10 min. The resulting slurry was stirred for 1 h, diluted with ether, filtered and the filtrate evaporated. The residual oil was dissolved in mL of methanol and treated with 0.5 mL of 4 M hydrogen chloride in dioxane. After stirring at room temperature for 1 h, the reaction mass was evaporated and the white solid recrystallized from ethyl acetate/hexane to give title compound, mp 146-148°C, 5.08 g, 91% yield.

55 WO 99/21564 PCT/US98/21750

C.

CO4ICIA20 S/

PO

3 Et 2 C(l).

0

OH

Br To a solution of 9-fluorenecarboxylic acid (Aldrich) g, 48 mmol) in THF (250 mL) at 0°C was added dropwise a solution of n-butyllithium (2.5M, 42.2 mL, 106 mmol) in THF. The yellow reaction was stirred at 0 0 C for 1 h, then (16.8 mL, 124 mmol) was added dropwise over 30 min. The reaction was stirred at 0oC for 30 min, then the reaction was warmed to RT for 30 h. The reaction was extracted with water (3 x 150 mL). The combined aqueous layers were extracted with ethyl ether (160 mL). The aqueous layer was made acidic with HC1 solution (IN, 100 mL), then extracted with dichloromethane (3 x 150 mL). The combined organic layers were dried over MgSO 4 Evaporation gave title compound as a white solid.

C(2) N

CF

3

H

Br To a solution of all of Part C(l) acid and DMF pIL) in CH 2 C1 2 (120 mL) under argon at O'C was added oxalyl 56 WO 99/21564 PCT/US98/21750 chloride (24 mL, 2.0 M in CH 2 C1 2 48 mmol) dropwise. The reaction was stirred at O'C for 10 min, then warmed to RT and stirred for 1.5 h. The reaction was concentrated in vacuo to give the crude acid chloride as a yellow oil. To a suspension of 2,2,2-trifluoroethylamine hydrochloride (6.51 g, 48 mmol) in CH 2 C1 2 (125 mL) at 0 0 C under argon was added triethylamine (16 mL, 115 mmol) followed by dropwise addition of a solution of the crude acid chloride in CH 2 C12 mL). The reaction was stirred at O'C for 1 h, diluted with CH 2 C1 2 (120 mL), and washed with water (2 x 100 mL), IN HC1 (2 x 100 mL), saturated NaHCO 3 (2 x 100 mL), and brine (2 x 100 mL), then dried over MgSO 4 Evaporation gave 17.6 g of an oil which was purified by flash chromatography on silica gel (500 The crude product was loaded in a mixture of CH 2 C1 2 and hexane, and eluted with a step gradient of 10% EtOAc/hexane (4L) to EtOAc/hexane (2L) to 20% EtOAc/hexane Pure fractions were combined and evaporated to give title compound (14.7 g, 72%) as a white solid m.p. 92-96°C).

C(3).

CF3 HN- CH3 A solution of 2.69 g (6.11 mmol) of Part C(2) compound in 15 mL of freshly distilled triethylphos-phite under argon was heated to 180°C for 16 h. The reaction was cooled and then the volatiles were distilled off at 100 0

C

at 1 Torr. The residue was triturated in ether to give title compound (2.25 g, 74%) as a white solid (mp 141- 143 0

C).

57 WO 99/21564 PCT/US98/21750 D. trans-9-[5-[2-Oxo-5-[ [[4'-(trifluoromethyl)[1,1'-biphenyl 2-yl]carbonyl] amino]- 1,3,2-dioxaphosphorinan-2-yl]pentyl]-N-(2,2,2trifluoroethyl)-9H-fluorene-9-carboxamide (Example 4) E. cis-9-[5-[2-Oxo-5-[[[4'-(trifluoromethyl)- [1,1'-biphenyl]-2-yl]carbonyl]amino]-1,3,2dioxaphosphorinan-2-yl]pentyl]-N-(2,2,2trifluoroethyl)-9H-fluorene-9-carboxamide (Example To a stirred solution of 1.68 g (3.42 mmol) of Part C compound in 10 mL of dichloromethane at room temperature under argon was added 1.35 mL (10.0 mmol) of bromotrimethylsilane. After 14 h, the reaction mixture was evaporated to provide a thick oil. The oil was dissolved in 10 mL of dichloromethane at room temperature under argon and treated with 0.91 mL (10.5 mmol) of oxalyl chloride and 50 JL of DMF. After 2 h, the reaction mixture was evaporated and redissolved in 10 mL of dichloromethane. To this solution at room temperature under argon, was added 1.09 g (3.2 mmol) of Part B compound and 1.0 mL (7.0 mmol) of triethylamine. After 1 h, the reaction was diluted with ethyl acetate and washed once with 10% citric acid solution, dried (MgSO,) and evaporated. Purification by flash chromatography on silica gel (5 x 20 cm column, 2 L 85:15 ethyl acetate/hexanes, then 2 L 3:97 methanol/ethyl acetate) provided two fractions.

The less polar fraction was designated Part D (Example 4) compound, white solid, mp 165-167 0 C, 805 mg, 34% yield.

MICROANALYSIS Calculated for C 3 8

H

3 5

F

6

N

2 0 5 P 0.38 H 2 0: C, 60.73; H, 4.80; N, 3.73; F, 15.17; P, 4.12 Found: C, 60.73; H, 4.70; N, 3.60; F, 15.33; P, 3.88.

MS (electrospray, ions) m/e 745.

58 WO 99/21564 PCT/US98/21750 The more polar fraction was designated Part E (Example compound, white foam, mp 110-114 0 C, 650 mg, 27%.

MICROANALYSIS Calculated for C 3 8

H

3 5

F

6

N

2 0 5 P 0.38 H 2 0: C, 60.73; H, 4.80; N, 3.73; P, 4.12 Found: C, 60.74; H, 4.78; N, 3.55; P, 3.84.

MS (electrospray, ions) m/e 745.

Example 6 trans-([ 2 -Oxo-2-[4-[9-[[(2,2,2-trifluoroethy)amino]carbonyl]-9H-fluoren-9-yl]butyl]- 1,3,2-dioxaphosphorinan-5-yl]carbamic acid, phenylmethyl ester.

H p

N

H

trans isomer A. N-Benzyloxycarbonyl-1,3-dihydroxy-2aminopropane H OH 0= OH To a stirred slurry of 1.00 g (4.24 mmol) of Example 5 Part A compound and 0.65 mL (4.7 mmol) of triethylamine in 10 mL of THF at 0 C under argon, was added dropwise, 0.6 mL (4.2 mmol) of benzyloxy carbonyl chloride (Aldrich) over min. The reaction was allowed to warm to room temperature and stirred for 14 h. The reaction mixture was diluted with ether and filtered. The filtrate was evaporated, the residue dissolved in 25 mL of methanol to which 2 drops of 4 N hydrogen chloride in dioxane was added 59 WO 99/21564 PCT/US98/21750 and the solution heated to 40 0 C for 30 min. Evaporation and trituration with hexane/ether gave title compound as a white solid, mp 104-1060C, 880 mg, 92% yield.

MS (electrospray, ions) m/z 226.

B. trans-[2-Oxo-2-[4-[9-[[( 2 2 2 -trifluoroethyl)amino]carbonyl]-9H-fluoren-9-yl]butyl]- 1,3,2-dioxaphosphorinan-5-yl]carbamic acid, phenvlmethyl ester To a stirred solution of 1.76 g (3.64 mmol) of Example 1 Part B compound in 5 mL of dichloromethane at room temperature under argon was added 1.65 mL (12.6 mmol) of bromotrimethylsilane. After 1 h, the reaction mixture was evaporated to provide a thick oil. The oil was dissolved in 20 mL of dichloromethane at room temperature under argon and treated with 1.2 mL (13.8 mmol) of oxalyl chloride and 100 pL of DMF. After 2 h, the reaction mixture was evaporated and redissolved in 5 mL of dichloromethane. To this solution at room temperature under argon, was added 820 mg (3.64 mmol) of Part A compound and 1.1 mL (7.9 mmol) of triethylamine. After 2 h, the reaction was diluted with ethyl acetate and washed once with 10% citric acid solution, dried (MgSOj) and evaporated. Purification by flash chromatography on silica gel (5 x 25 cm column, 85:15 ethyl acetate/hexane) provided title compound, white solid, mp 160-163°C, 535 mg, 23% yield.

MS (electrospray, ions) m/e 617.

60 WO 99/21564 PCT/US98/21750 Example 7 trans-1-(Phenylmethyl)-N-[2-[4-[9-[[(2,2,2-trifluoroethyl)amino]carbonyl]-9Hfluoren-9-yl]butyl]-1, 3 2 -dioxaphosphorinan-5-yl]-2-piperidinecarboxamide, hydrochloride.

0

H

0 0 trans isomer A. N-Benzylpipecolic acid hydrochloride CO2H

*HCI

A mixture of ethyl pipecolinate hydrochloride (10 g, 52 mmol), benzylbromide (7 mL, 57 mmol) and potassium carbonate (15 g, 114 mmol) in DMF (80 mL) was stirred at room temperature overnight. The solvent was removed in vacuo. The residue was partitioned between dichloromethane (100 mL) and water (50 mL). The aqueous layer was extracted with dichloromethane (2 x 100 mL), dried over Na 2

SO

4 then evaporated to give a yellow oil. The crude product was chromatographed (500 g silica gel) eluting with a stepgradient of 8%-15% ethyl acetate in hexane. Pure fractions were combined to give title compound (12.3 g, 96%) as a colorless oil.

B. trans-l-(Phenylmethyl)-N-[2-[4-[9- 2 ,2,2-trifluoroethyl)amino]carbonyl]-9Hfluoren-9-yl] butyl] -1,3,2-dioxaphosphorinan-5- Y11 2 -Diperidinecarboxamide, hydrochloride An argon-purged slurry of 511 mg (0.829 mmol) of Example 6 compound and 100 mg of 10% palladium-on-charcoal in 5 mL of ethanol at room temperature was partially evacuated and subjected to hydrogenation from a filled balloon. After 16 h, the reaction mixture was purged with 61 WO 99/21564 PCT/US98/21750 argon and filtered through a 0.45 g nylon filter.

Evaporation and then re-evaporation from dichloromethane gave a white foam. This material was dissolved in 10 mL of dichloromethane and stirred under argon at room temperature while 212 mg (0.83 mmol, HC1 salt) of Part A compound, 125 mg (0.83 mmol) of HOBt, 175 mg (0.92 mmol) of EDCI and 175 gL (1.26 mmol) of triethylamine were added. After 6 h, the reaction mixture was quenched with saturated sodium bicarbonate solution and extracted twice with ethyl acetate. The organic extracts were combined, dried (MgSO 4 and evaporated. Purification by flash chromatography on silica gel (5 x 20 cm column, ethyl acetate) provided the free base of title compound. This material was dissolved in dichloromethane, treated with 0.3 mL of 4 M hydrogen chloride in dioxane and evaporated to provide title compound as a white solid, mp 125-128 0 C, 365 mg, 61% yield.

MICROANALYSIS Calculated for C 3 6H 4 1

F

3

N

3 0 5 P+HCl+1 dioxane+0.33

H

2 0: C, 59.01; H, 6.27; N, 5.16; Cl, 4.35; F, 7.00 Found: C, 59.00; H, 6.25; N, 5.11; Cl, 4.14; F, 6.96.

MS (electrospray, ions) m/e 684.

Example 8 trans-9-[4-[2-Oxo-5-[[2-(2-pyridinyl)benzoyl]amino]-1,3,2-dioxaphosphorinan- 2-yl]butyl-N-(2,2,2-trifluoroethyl)-9H-fluorene-9-carboxamide, hydrochloride.

rl o 0 trans isomer 62 WO 99/21564 PCT/US98/21750 A. 2-(2-Pyridyl)benzoic acid o

N

0

I

HO

2-Methyl-1-(2-pyridyl)benzene

-N

To a degassed solution of 2-bromopyridine (1.9 mL, mmol) in ethylene glycol dimethyl ether (60 mL) under argon, tetrakis(triphenylphosphane) palladium (700 mg, 0.6 mmol) was added. After stirring for 10 min, 2methylphenyl boronic acid (2.9 g, 22 mmol) was added, followed by sodium bicarbonate solution (5.04 g, 60 mmol in mL water). The mixture was heated to reflux 85 0

C)

and stirred overnight. After cooling to room temperature, the solvent was evaporated, the residue was partitioned between water and ether, and the aqueous layer was extracted twice with ether. The combined organic layers were dried (Na 2

SO

4 and the solvent was evaporated to give a black oil. This oil was distilled at <1 Torr at -95 0 C to give title compound (2.75 g, 82% yield) as a clear oil.

A(2).

COOH

A solution of Part A(l) compound (850 mg, 5.0 mmol) and potassium permanganate (1.9 g, 12.0 mmol) in water mL) was heated to reflux and stirred for 1 hour. The hot 63 WO 99/21564 PCT/US98/21750 reaction mixture was filtered, and the filtrate was evaporated to dryness. The solid residue was dissolved in water (5 mL) and acidified with acetic acid to pH 4-5. The resulting precipitate was isolated by filtration and rinsed with water to give a white solid (800 mg) which was recrystallized from hot ethanol (12 mL) to give title compound as a white solid (453 mg, 45% yield).

B. trans-9-[4-[2-Oxo-5-[[2-(2-pyridinyl)benzoyl]amino]-1,3,2-dioxaphosphorinan-2yl]butyl]-N-(2,2,2-trifluoroethyl)-9Hfluorene-9-carboxamide, hydrochloride An argon-purged slurry of 505 mg (0.82 mmol) of Example 6 compound and 100 mg of 10% palladium-on-charcoal in 10 mL of methanol at room temperature was partially evacuated and subjected to hydrogenation from a filled balloon. After 4 h, the reaction mixture was purged with argon and filtered through a 0.45 g nylon filter.

Evaporation and then re-evaporation from toluene gave a white foam. This material was dissolved in 10 mL of dichloromethane and stirred under argon at room temperature while 169 mg (0.83 mmol) of Part A compound, 162 mg (0.85 mmol) of EDCI and 60 pL (0.43 mmol) of triethylamine were added. After 16 h, the reaction mixture was quenched with saturated sodium bicarbonate solution and extracted twice with ethyl acetate. The organic extracts were combined, dried (Na 2

SO

4 and evaporated. Purification by flash chromatography on silica gel (5 x 15 cm column, 1:24 methanol/ethyl acetate) provided the free base of the title compound. This material was dissolved in dichloromethane, treated with 0.3 mL of 4 M hydrogen chloride in dioxane and evaporated to provide title compound as a white solid, mp 132-136°C, 524 mg, 81% yield.

64 WO 99/21564 WO 9921564PCTJUS98/21 750 MICROANALYSIS Calculated for C 35

H

3 3

F

3

N

3 0 5 P HCl dioxane+ 0.25 H 2 0: C, 59.09; H, 5.40; N, 5.30; Cl, 4.47; F, 7.19; P, 3.91 Found: C, 59.12; H, 5.28; N, 5.24; Cl, 4.48; F, 7.37; P, 4.08.

MS (electrospray, ions) m/e 664.

Examole 9 trans-9-[4-[5-[[2-(2-Benzothiazolyl)benzoyl~amino]-2-oxo-1 3,2dioxaphosphorinan-2-yI]buy-N-(2,2,2trfluoroethy)-9H-fluorene- 9-carboxamide.

0 N

QN

H

trans isomer

A.

H

0 Part A compound is a commercial compound supplied by Maybridge Chemical Company.

B. trans-9- (2-Benzothiazolyl) benzoyl] amino] -2-oxo-l, 3, 2-dioxaphosphorinan-2-yl ]butyl] 2-trifluoroethyl) -9H-fluorene-9-carboxamide An argon-purged slurry of 371 mg (0.60 mmol) of Example 6 compound and 90 mg of 10% palladium-on-charcoal in 10 mL of methanol at room temperature was partially evacuated and subjected to hydrogenation from a filled balloon. After 4 h, the reaction mixture was purged with argon and filtered through a 0.45 g. nylon filter.

Evaporation and then re-evaporation from toluene gave a 65 WO 99/21564 PCT/US98/21750 white foam. This material was dissolved in 5 mL of dichloromethane and stirred under argon at room temperature while 153 mg (0.60 mmol) of Part A compound, 114 mg (0.60 mmol) of EDCI and 42 iL (0.3 mmol) of triethylamine were added. After 16 h, the reaction mixture was quenched with saturated sodium bicarbonate solution and extracted twice with ethyl acetate. The organic extracts were combined, dried (Na 2

SO

4 and evaporated. Purification by flash chromatography on silica gel (5 x 15 cm column, 1:3 hexanes/ethyl acetate) provided title compound as a white solid, mp 117-118 0 C, 269 mg, 62% yield.

MICROANALYSIS Calculated for C 37

H

33

F

3

N

3 0 5

PS:

C, 61.75; H, 4.62; N, 5.84; F, 7.92; S, 4.45; P, 4.30 Found: C, 62.02; H, 4.97; N, 5.55; F, 7.64; S, 4.06; P, 4.42.

MS (electrospray, ions) m/e 720.

Example trans-9-[4-[5-[[2-(4-Morpholinyl)benzoyl]amino]-2-oxo-1,3,2-dioxaphosphorinan- 2 -yl]butyl]-N-(2,2,2-trifluoroethyl)-9H-fluorene-9-carboxamide.

N

H

0 trans isomer A. 2-(l-Morpholino)benzoic acid C0

HO

66 WO 99/21564 PCT/US98/21750 Methyl 2 -(l-morpholino)benzoate 0 crCOOCH3 A solution of methyl 2 -fluorobenzoate (15.4 g, 100 mmol) in morpholine (44 mL, 500 mmol) was heated at for 30 min, and then warmed to 100 0 C and stirred for 2 h, then cooled to 50°C and stirred overnight. The reaction was then heated to reflux for 2.5 h. The excess morpholine was evaporated. The remainder was dissolved in ethyl acetate and washed successively with H 2 0 (50 mL), saturated NaHCO 3 solution (2 x 50 mL), H 2 0 (3 x 50 mL) and brine mL). Drying (MgS0 4 and evaporation gave a yellow oil.

The crude product was dissolved in dichloromethane/ethyl acetate/hexane and chromatographed (400 g silica gel) eluting with a step gradient of 20% to 35% ethyl acetate in hexane. Pure fractions were combined and evaporated to give title compound (10.5 g, 48%) as an oil which crystallized on standing to a white solid.

2-(l-Morpholino)benzoic acid 0

HO/--O

Sodium hydroxide (10 g, 250 mmol) was dissolved in water (75 mL) and added to a solution of Part A(l) compound (10.4 g, 47.1 mmol) in methanol (75 mL). The reaction was stirred at RT for 1 h and the solvent was evaporated. The white residue was dissolved in H 2 0 (100 mL) and adjusted to pH 1.5 with IN HC1. The aqueous layer was extracted with chloroform (3 x 250 mL). The combined organic layers were 67 WO 99/21564 PCT/US98/21750 dried (Na 2

SO

4 and evaporated to give title compound (9.76 g, 85%) as a white solid 156-157 0

C).

B. trans-9-[4-[5-[[2-(4-Morpholinyl)benzoyl]amino] -2-oxo-1, 3,2-dioxaphosphorinan- 2-yl]butyl]-N-(2,2,2-trifluoroethyl)-9Hfluorene-9-carboxamide An argon-purged slurry of 359 mg (0.582 mmol) of Example 6 compound and 100 mg of 10% palladium-on-charcoal in 15 mL of methanol at room temperature was partially evacuated and subjected to hydrogenation from a filled balloon. After 16 h, the reaction mixture was purged with argon and filtered through a 0.45 g nylon filter.

Evaporation and then re-evaporation from toluene gave a white foam. This material was dissolved in 5 mL of dichloromethane and stirred under argon at room temperature while 121 mg (0.584 mmol) of Part A compound, 88 mg (0.59 mmol) of HOBt, 122 mg (0.64 mmol) of EDCI and 41 gL (0.3 mmol) of triethylamine were added. After 6 h, the reaction mixture was quenched with saturated sodium bicarbonate solution and extracted twice with ethyl acetate. The organic extracts were combined, dried (Na 2

SO

4 and evaporated. Purification by flash chromatography on silica gel (5 x 20 cm column, ethyl acetate) provided title compound as a white solid, mp 108-111 0 C, 265 mg, 66% yield.

MICROANALYSIS Calculated for C 34

H

37

F

3

N

3 0 6 P+0.15 H 2 0+0.23 EtOAc: C, 60.38; H, 5.68; N, 6.05; F, 8.21 Found: C, 60.37; H, 5.54; N, 5.93; F, 7.91.

MS (electrospray, ions) m/e 695.

68 WO 99/21564 PCT/US98/21750 Example 11 trans-9-[4-[5-t[2-(2-Benzoxazolyl)benzoyl]amino]-2-oxo-1,3,2-dioxaphosphorinan- 2-yl]butyl]-N-(2,2,2-trifluoroethyl)-9H-fluorene-9-carboxamide.

H CF

N

o'a 0

N

H

A. 2 -(2-Bromophenyl)benzoxazole Br To a stirred slurry of 5.46 g (50.0 mmol) of 2aminophenol (Aldrich) and 10.05 g (50.0 mmol) of 2bromobenzoic acid in 100 mL of xylenes under argon at room temperature was added 6.10 g (100 mmol) of boric acid.

Using a Dean-Stark trap, the reaction was heated to reflux for 48 h. A total of 3.2 mL of water was separated. The reaction mixture was cooled, diluted with ethyl acetate and filtered. The filtrate was washed once with 3 M hydrochloric acid, once with water and three times with saturated sodium bicarbonate solution. The organic phase was dried (MgSOJ) and evaporated. Purification by flash chromatography on silica gel (5x20 cm column, 2:1 dichloromethane/hexane) gave title compound as a light yellow solid, 5.90 g, 43%.

69 WO 99/21564 PCT/US98/21750 B. 2 2 -Benzoxazolyl)benzoic acid

CO

2

H

To a stirred solution of 700 mg (2.55 mmol) of Part A compound in 10 mL of THF under argon at -78°C was added a solution of t-butyllithium (3.3 mL, 1.7 M, 5.6 mmol) in pentane over the course of 30 min. The reaction mixture was stirred for 1 h and then a dry stream of carbon dioxide gas was passed through the solution for 1h. The reaction was allowed to warm to room temperature in situ. After 16 h, the mixture was diluted with ether and washed once with mL of 10% sodium hydroxide solution. The aqueous phase was adjusted to pH 3.5 using solid citric acid. The resulting solid was filtered, washed with water and airdried to give 610 mg of title compound as a pink solid, mp >250 0 C, 100%.

C. trans-9-[4-[5-[[ 2 -(Benzoxazolyl)benzoyl]amino]-2-oxo-l,3,2-dioxaphosphorinan-2yl)butyl]-N-(2,2,2-trifluoroethyl)-9Hfluorene-9-carboxamide To a stirred solution of 117 mg (0.24 mmol) of Example 6 compound under argon at room temperature in 3 mL of dichloromethane was added 72 mg (0.30 nmol) of Part B compound, 45 mg (0.3 mmol) of HOBt, 65 mg (0.30 mmol) of EDCI and 20 gL (0.15 mmol) of triethylamine. After 16 h, the reaction mixture was quenched with saturated sodium bicarbonate solution and extracted twice with ethyl acetate. The organic extracts were combined, dried (Na 2

SO

4 and evaporated. Purification by flash chromatography on silica gel (2.5 x 15 cm column, 1:3 hexanes/ethyl acetate) provided title compound as a white solid, mp 104-107°C, 38 mg, 20% yield.

70 Wo 99/21564 PCT/US98/21750 MICROANALYSIS Calculated for C 37

H

33

F

3

N

3 0 6 P+2.3 H 2 0+0.5 EtOAc: C, 59.36; H, 5.31; N,5.32; F, 7.22; P, 3.92 Found: C, 59.32; H, 5.24; N,5.43; F, 7.45; P, 4.20.

MS (electrospray, ions) m/e 704.

Following are examples of additional compounds of the invention which may be prepared employing procedures described hereinbefore and in Examples 1 to 11. Although the following compounds are shown in the form of cisisomers, in accordance with the invention, the corresponding trans-isomers are covered as well.

o

R

2 O A B NCOR1 H 0

F

3 C-0N 0 ^N

H

S F3 HN 0

H

71 wo 99/21564 WO 9921564PCT[US98/2 1750 72 WO 99/21564 WO 9921564PCT/US98/21 750 IHCONHEt 73 WO 99/21564 WO 9921564PCTIUS98/21 750 74 WO 99/21564 WO 9921564PCT/US98/2 1750 75

Claims (14)

1. A compound which has the structure 0 R2 L A B, I S. S S 4* including-or pharmaceutically acceptable salts thereof, or N-oxides thereof, wherein A is a bond or -N- R where R 5 is H or lower alkyl, or R 5 together with R 2 forms a carbocyclic or heterocyclic ring system containing 4 to 8 members in the ring; B is a fluorenyl-type group of the structure RR 4 r R 3 R3 R4' I or Het N Waut1 Het2 B is an indenyl-type group of the structure W:\fionaNKI\Marked Up\10876.doc WO 99/21564 WO 9921564PCT/US98/21 750 or R RR3b or (a 2,3 or 4) R 3 a R 3 R Tr R 3 b orR3 R 3 1 (CH 2 )a Ra Ra and Rb may be the same or different and are hydrogen, al.kyl, aryl, arylalkyl or heteroaryl linked to the ring via a carbon atom. R 5 a is H, lower alkyl or aryl; R 1 is independently alkyl, alkenyl, alkynyl, aryl, alkoxy, aryloxy, arylalkoxy, heteroaryl, heteroarylalkoxy, arylalkyl, heteroarylalkyl, cycloalkyl, cycloalkylalkyl, polycycloalkyl, polycycloalkylalkyl, cycloalkenyl, cycloheteroalkyl, heteroaryloxy, cycloalkenylalkyl, polycycloalkenyl, polycycloalkenylalkyl, heteroarylcarbonyl, amino, alkylamino, arylamino, heteroarylamino, cycloalkyloxy, cycloalkylanino, all optionally substituted through available carbon atoms with 1, 2, 3 or 4 groups selected from hydrogen, halo, alkyl, halo-alkyl, alkoxy, haloalkoxy, alkenyl, alkynyl, cyclo- alkyl, cycloalkylalkyl, cycloheteroalkyl, cycloheteroalkyl- alkyl, 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, aminocarbonyl, 77 WO 99/21564 WO 9921564PCT/US98/21 750 alkynylaminocarbonyl, alkylaminocarbonyl, alkenylamino- carbonyl, alkylcarbonyloxy, arylcarbonyloxy, alkylcarbonylamino, arylcarbonylamino, arylsulfinyl, arylsulfinylalkyl, arylsulfonyl, alkylsulfonyl, arylsulfonylanino, heteroarylcarbonylamino, heteroarylsulfinyl, heteroarylthio, heteroarylsulfonyl, alkylsulfinyl; or RI and R 5 a can be joined to form a ring of the structure R 0 0 R21 JjN- R2 where a is: CE23, -CH-C- or -CC- 11 9 1 1 1 1 0 R 24 R 2 S R 24 R 2 R 23 R 24 and R 2 5 are independently hydrogen, alkyl, alkenyl, alkynyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, cycloalkyl, or cycloalkylalkyl; R 2 0 R 2 1 R 2 2 are independently hydrogen, halo, alkyl, alkenyl, alkoxy, aryloxy, aryl, arylalkyl, alkylmercapto, arylmercapto, cycloalkyl, cycloalkylalkyl, heteroaryl, heteroarylalkyl, hydroxy or haloalkyl; and these substituents may either be directly attached to R 1 or attached via an alkylene at an open position; R 2 is hydrogen, alkyl, alkenyl, alkynyl, haloalkyl, alkoxyl, (alkyl or aryl) 3 Si (where each alkyl or aryl. group is independent), cycloalkyl, cycloalkenyl, substituted alkylamino, substituted arylalkylamino, aryl, arylalkyl, arylamino, aryloxy, heteroaryl, heteroarylamino, heteroaryloxy, arylsulfonylamino, heteroarylsulfonylanino, arylthio, arylsulfinyl, arylsulfonyl, alkylthio, alkylsulfinyl, alkylsulfonyl, heteroarylthio, heteroarylsulfinyl, heteroarylsulfonyl, cycloheteroalkyl, cycloheteroalkylalkyl, -P0(R 13 (R1 4 (where R 1 3 and R 1 4 are independently alkyl, aryl, alkoxy, aryloxy, heteroaryl, heteroarylalkyl, heteroaryloxy, heteroarylalkoxy, cycloheteroalkyl, cycloheteroalkylalkyl, cycloheteroalkoxy, 78 79 or cycloheteroalkylalkoxy); cyano, 1,1-alkoxyl or aryloxy) 2 alkyl (where the two aryl or alkyl substituents can be independently defined), R 2 may be substituted with 1, 2, 3 or 4 substituents, which can be any of substituents for R 1 or haloalkylamino, alkylamino, cycloalkylamino, arylamino, heteroarylamino, alkoxyamino, aryloxyamino, heteroaryloxylamino, heterocyclylamino (where the heterocycle is connected to the carbonyl group via a nitrogen or carbon atom); L 1 is a linking group containing from 1 to 10 carbons in a linear chain inc dking comprising alkylene, alkenylene or alkynylene, which may contain, within the linking chain any of the following: one or two alkenes, one or two alkynes, an oxygen, an amino group, an oxo group, and may be substituted with one to five alkyl or halo groups; L 2 may be the same or different from L 1 and may independently be any of the Ll groups set out above or a single bond; R 3 R 3 R 4 and R 4 may be the same or different and are independently 15 selected from H, halogen, CF 3 haloalkyl, hydroxy, alkoxy, alkyl, aryl, alkenyl, alkenyloxy, alkynyl, alkynyloxy, alkanoyl, nitro, amino, thiol, alkylthio, alkylsulfinyl, alkylsulfonyl, carboxy, alkoxycarbonyl, aminocarbonyl, alkylcarbonyloxy, alkylcarbonylamino, cycloheteroalkyl, cycloheteroalkylalkyl, cyano, Ar-, Ar-alkyl, ArO, Ar-amino, Ar-thio, Ar-sulfinyl, Ar-sulfonyl, Ar-carbonyl, Ar-carbonyloxy or Ar- carbonylamino, wherein Ar is aryl or heteroaryl and Ar may optionally include 1, 2 or 3 additional rings fused to Ar; R 3a and R 3 b are the same or different and are independently any of the R 3 groups; 8 and H W:\fiona\NKIMarked Up\l0876.doc WO 99/21564 PCT/US98/21750 are the same or different and independently represent a or 6 membered heteroaryl ring which contains 1, 2, 3 or 4 heteroatoms in the ring which are independently N, S or 0; and including N-oxides; X is a bond, or is one of the following groups: -s- -N- R 6 R7\ R 8 c\ o- o R 9 R 0 R 9 R 1 0 c=c-- I I R 9 RI" or Y- R9 RIO wherein Y is 0, N-R 6 or S; n' is 0, 1 or 2; R 6 is H, lower alkyl, aryl, -C(0)-R 1 1 or -0-R 1 1 R 7 and R 8 are the same or different and are independently H, alkyl, aryl, halogen, -0-R 1 2 or R 7 and R 8 together can be oxygen to form a ketone; R 9 R 1 0 R 9 and R 1 0 are the same or different and are independently H, lower alkyl, aryl or -0-R 11 R 9 and R 1 0 are the same or different and are independently H, lower alkyl, aryl, halogen or -0-R 11 R 11 is alky or aryl; R 12 is H, alkyl or aryl; 80 WO 99/21564 PCTIUS98/21750 with the proviso that when A is a bond, R 2 L 2 cannot be H.

2. The compound as defined in Claim 1 wherein A is a bond.

3. The compound as defined in Claim 1 wherein A is

4. The compound as defined in Claim 1 wherein A is R s The compound as defined in Claim 1 wherein B is a fluorenyl-type group.

6. The compound as defined in Claim 1 wherein B is an indenyl-type group.

7. The compound as defined in Claim 1 wherein B is x A is NH; X is a bond, oxygen or sulfur; R 3 and R 4 are the same or different and are H or F; R 1 is aryl, phenyl, biphenyl or cycloheteroalkyl; R 2 is aryl, phenyl, heteroaryl, imidazolyl, pyridyl, cyclohexyl, PO(R 13 (R 14 heteroarylthio, benzthiazole-2- thio, imidazole-2-thio, alkyl, alkenyl or l,3-dioxan-2-yl, wherein each of the above is optionally substituted; L 1 is a chain containing 1 to 5 atoms in a linear chain; L 2 is a bond or lower alkylene.

8. The compound as defined in Claim 1 which is 9 -[4-[2-oxo-5-[[[4'-(trifluoromethyl)[1,1'-biphenyl]-2- yl] carbonyl] amino] -1,3,2-dioxaphosphorinan-2-yl]butyl] -N- (2,2,2-trifluoroethyl)-9H-fluorene-9-carboxamide; (Z)-9-[4-[2-oxo-5-[[[4'-(trifluoromethyl)-[1,1'- biphenyl] -2-yl] carbonyl] amino] -1,3,2-dioxaphosphorinan-2- yl]butyl] (2,2,2-trifluoroethyl)-9H-fluorene-9- carboxamide; 81 WO 99/21564 WO 9921564PCTIUS98I2 1750 trans-9- (benzoylamino) -2-oxo-1, 3,2- dioxaphosphorinan-2-yllbutyjj 2-trifluoroethyl) -9H- fluorene-9-carboxamide; trans-9- [2-oxo-5- (trifluoromethyl) biphenyl] -2-yl] carbonyl] amino] 2-dioxaphosphorinan-2- yllipentyl] 2-trifluoroethyl) -9H-fluorene-9- carboxamide; cis-9- [2-oxo-5- (trifluoromethyl) biphenyl] -2 -yl] carbonyl] amino] -1,3,2 -dioxaphosphorinan-2 yllpentyl] 2-trifluoroethyl) -9H-fluorene-9- carboxamide; trans-[2-oxo-2-[4-[9-[ [(2,2,2-trifluoroethyl)- amino] carbonyl] 9H-fluoren-9-yllbutyl] 2-dioxa- carbamic acid; trans-l-(phenylmethyl)-N-[2-[4-[9-[ trifluoroethyl) amino] carbonyl] -9H-fluoren-9-yl] -butyl] 1,3, 2-dioxaphosphorinan-5-yl] 2 -piperidinecarboxamide; trans-9- [2-oxo-5- (2-pyridinyl)benzoyl] amino] 3,2-dioxaphosphorinan-2-yllbutyl-N..(2,2,2- trifluoroethyl) -9H-fluorene-9-carboxamide; trans-9- (2-benzothiazolyl)benzoyllamino] 2-oxo-1, 3, 2-dioxaphosphorinan-2-yljbutyl] (2,2,2- trifluoroethyl) -9H-fluorene-9-carboxamide; trans-9- (4-morpholinyl)benzoyllamino] -2- oxo-1, 3, 2-dioxaphosphorinan-2-yllbutyl] (2,2,2- trifluoroethyl) -9H-fluorene-9 -carboxaiide; trans-9- 2 -benzoxazolyl)benzoyllamino] -2- oxo-1, 3, 2-dioxaphosphorinan-2-yl]butyl] (2,2,2- trifluoroethyl) -9H-fluorene--9-carboxamide or an ester thereof or an N-oxide thereof, or a pharmaceutically acceptable salt thereof.

9. The compound as defined in Claim 1 having the structure 82 wo 99/21564 WO 9921564PCT[US98/2 1750 R'CO-= n geometry R'CO-= n geometry i CF 3 CO cI CF 3 No trans C6H~ (co I cis 1 trans 2 trans 2 cis 1 trans 1 trans 1 trans S leN 0 &co trans 1 trans 0 11 OC- N trans 83 WO 99/21564 WO 9921564PCTIUS98/21 750

11. The compound as defined in Claim 1 having the name 9 4 2 -oxo-5-[[[4'-(trifluoromethyl).[llu1 biphenyl] -2-yl] carbonyl] amino] 2-dioxa-phosphorinan-2- yllbutyl] 2-trifluoroethyl) -9H-fluorene-9- carboxamide; trans-9- (benzoylanino) -2-oxo-1, 3,2- dioxaphosphorinan-2 -yl] butyl] 2-trifluoro-ethyl) -9H- fluorene-9-carboxamide; trans-9- [2-oxo-5- (trifluoromethyl) biphenyl] -2-yl] carbonyl] amino] 2-dioxaphosphorinan-2- yllpentyl] 2-trifluoroethyl) -9H-fluorene-9- carboxamide; trans- [2-oxo-2- 2-trifluoroethyl) amino] carbonyl] 9H-fluoren-9-yllbutyl] 2-dioxa- phosphorinan-5-yllcarbamic acid; trans-i- (phenylmethyl) trifluoroethyl) amino] carbonyl] -9H-fluoren-9-yllbutyl] 1,3, 2-dioxaphosphorinan-5-yl] -2-piperidinecarboxamide; trans-9- [2-oxo-5- (2-pyridinyl)benzoyl] amino] 3,2-dioxaphosphorinan-2-yllbutyl] (2,2,2- trifluoroethyl) -9H-fluorene-9-carboxamide; trans-9-[4-[5-[ (2-benzothiazolyl)benzoyllamino]- 2-oxo-1, 3,2-dioxaphosphorinan-2-yl]butyl] (2,2,2- trifluoroethyl) -9H-fluorene-9-carboxamide; trans- 9 -[4-[5-[[2-(4-morpholinyl)benzoylaino2 oxo-1, 3, 2-dioxaphosphorinan-2-yllbutyl] (2,2,2- trifluoroethyl) 9 H-fluorene-9-carboxamide; trans-9-[4-[5-[ 2 2 -benzoxazolyl)benzoyl]amino] -2- oxo-l, 3, 2-dioxaphosphorinan-2-yllbutyl] (2,2,2- trifluoroethyl)-9H-fluorene-9-carboxamide; or an N-oxide thereof, or pharmaceutically acceptable salts thereof or esters thereof.

12. The compound as defined in Claim 1 wherein A is NH and R 2 L 2 is CF 3 CH 2

13. A method for preventing, inhibiting or treating atherosclerosis, pancreatitis hyperglycemia or obesity in a maimmalian species, which comprises administering to a 84 WO 99/21564 PCT/US98/21750 patient in need of treatment a therapeutically effective amount of a compound as defined in Claim 1.

14. A method of lowering serum lipid levels, cholesterol and/or triglycerides, or inhibiting and/or treating hyperlipemia, hyperlipidemia, hyperlipoproteinemia, hypercholesterolemia and/or hypertriglyceridemia, and/or preventing, inhibiting or treating atherosclerosis, pancreatitis or obesity in a mammalian species, which comprises administering to a patient in need of treatment a therapeutically effective amount of a compound as defined in Claim 1. The method as defined in Claim 14 where B is R3 R 4 A is NH; X is a bond, oxygen or sulfur; R 3 and R 4 are the same or different and are H or F; R 1 is aryl, phenyl, biphenyl or cycloheteroalkyl; R 2 is aryl, phenyl, heteroaryl, imidazolyl, pyridyl, cyclohexyl, PO(R 13 (R 14 heteroarylthio, benzthiazole-2- thio, imidazole-2-thio, alkyl or alkenyl, 1,3-dioxan-2-yl, wherein each of the above is optionally substituted; L 1 is a chain containing 1 to 5 atoms in a linear chain; L 2 is a bond or lower alkylene.

16. The method as defined in Claim 13 where B is R 3 R 4 0 0 X is a bond, oxygen or sulfur; R 3 and R 4 are the same or different and are H or F; 85 86 R 2 is aryl, phenyl, heteroaryl, imidazolyl, pyridyl, cyclohexyl, PO(R 13 14 heteroarylthio, benzthiazole-2-thio, imidazole-2-thio, al kyl or alkenyl, 1 ,3-d ioxan-2- yI, wherein each of the above is optionally substituted; R1 is phenyl, biphenyl or piperidine; Ll is a chain containing 1 to 5 atoms in a linear chain; L 2 is a bond or lower alkylene.

17. A compound according to claim 1 substantially as hereinbefore described with reference to any of the examples. DATED: 15 January, 2001 PHILLIPS ORMONDE FITZPATRICK 15 Attorneys for: BRISTOL-MYER SQUIBB COMPANY 0 00* Wfon*\NI\Mrkod Up\10876.doo