Note: Descriptions are shown in the official language in which they were submitted.
CA 02307889 2000-04-27
WO 99/21564 PCT1US98/21750
CYCLIC PHOSPHONATE ESTER INHIBITORS OF
MICROSOMAL T.~GLYCERIDE 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.
Bac)~,around of the invention
The microsomal triglyceride transfer protein (MTP)
catalyzes the transport of triglyceride (TG), cholesteryl
ester (CE), and phosphatidylcholine (PC) between small
unilamellar vesicles (SW). Wetterau & Zilversmit, Chem.
Phys. Lipids ~$, 205-22 (1985). When transfer rates are
expressed as the percent of the donor lipid transferred per
time, MTP expresses a distinct preference for neutral lipid
transport (TG and CE), relative to phospholipid transport.
The protein from bovine liver has been isolated and
characterized. Wetterau & Zilversmit, Chem. Phys. Limas
_3$, 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 a ., J. Biol. Chem.
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9800-7 (1990). The presence of PDI in the transfer protein
is supported by evidence showing that (1) the amino
terminal 25 amino acids of the bovine 58,000 kDa component
of MTP is identical to that of bovine PDI, and (2)
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 ,3~5, 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 4-
hydroxylase. Koivu et~, J. Biol. Chem. 2
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 etal., Biochemistry ,~(~, 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. Bionhys. Acta 87~, 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
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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 Diseasg, 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. ~, B9-12
(1988). A link between the apoB gene and
abetalipoproteinemia has been excluded in several families.
Talmud et al., ~T. Clin. Invest. ~, 1803-6 (1988) and Huang
et a ., ,Am. J. Hum. Gent. 4~C, 1141-8 (2990).
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 .
,~ vit , MTP catalyzes the transport of lipid
molecules between phospholipid membranes. Presumably, it
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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,
Bioc~em. Biqphvs. Acta 87~, 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 ., J. Biol. Chem.
~, 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. ~?, 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, ~T. Lipid Res. ,~5, 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,
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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 DC2lb)) 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 irk vivo. This Canadian patent application discloses
a method for identifying the MTP inhibitors
(\
N N
/
which has the name 2-[1-(3,3-diphenylpropyl)-4-
piperidinyl]-2, 3-dihydro-3-oxo-1~i-isoindole hydrochloride
and
O
\ ~N \
/ ,, J OCH3
which has the name 1-[3-(6-fluoro-1-tetralanyl)-methyl]-4-
O-methoxyphenyl piperazine.
EP 0643057A1 published March 15, 1995, discloses MTP
inhibitors of the structure
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I
or
II
O
R3. I- / _N~N_ R~
X '
R4
0
RS
R N ~N R'
or
III
o
R7
. _ J.
Y
where X is : CHIie, -cH- C8 Or -C= c- :
I I
R9 Ri0 R9 Ri0
R8, R9 and R1~ are independently hydrogen, alkyl, alkenyl,
alkynyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl,
cycloalkyl, or cycloalkylalkyl;
Y is -(cHZ)~; or -iC-
O
where m is 2 or 3;
R1 is alkyl, alkenyl, alkynyl, aryl, heteroaryl,
arylalkyl (wherein alkyl has at least 2 carbons),
diarylalkyl, arylalkenyl, diarylalkenyl, arylalkynyl,
diarylalkynyl, diarylalkylaryl, heteroazylalkyl (wherein
alkyl has at least 2 carbons), cycloalkyl, or
cycloalkylalkyl (wherein alkyl has at least 2 carbons); all
of the aforementioned R1 groups being optionally
substituted through available carbon atoms with l, 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
R1 is a group of the structure
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Rts
'~~ _
,
tt
Z
R~2
R~3 ~~ Rta
Rll is a bond, alkylene, alkenylene or alkynylene of
up to 6 carbon atoms, arylene (for example
)
~ ,
or mixed arylene-alkylene (for example
~- ( CH1 ) a. )
where n is 1 to 6;
R12 is hydrogen, alkyl, alkenyl, aryl, heteroaryl,
haloalkyl, arylalkyl, arylalkenyl, cycloalkyl, aryloxy,
alkoxy, arylalkoxy, heteroarylalkyl or cycloalkylalkyl;
Z is a bond, O, S, N-alkyl, N-aryl, or alkylene or
alkenylene of from 1 to 5 carbon atoms;
R13~ R14~ R15~ ~d R16 are independently hydrogen,
alkyl, halo, haloalkyl, aryl, cycloalkyl, cycloheteroalkyl,
alkenyl, alkynyl, hydroxy, alkoxy, nitro, amino, thio,
alkylsulfonyl, arylsulfonyl, alkylthio, arylthio, carboxy,
aminocarbonyl, alkylcarbonyloxy, alkylcarbonylamino,
arylalkyl, heteroaryl, heteroarylalkyl, or aryloxy;
or R1 is
Ri7
(C8Z)D~
Ri8
wherein p is 1 to 8 and R1~ and R18 are each independently
H, alkyl, alkenyl, aryl, arylalkyl, heteroaryl,
heteroarylalkyl, cycloalkyl or cycloalkylalkyl, at least
one of R1~ and R18 being other than H;
or Rl is
CA 02307889 2000-04-27
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R20
Ri9
RZi
wherein R19 is aryl or heteroaryl;
R2~ is aryl or heteroaryl;
R21 is H, alkyl, aryl, alkylaryl, arylalkyl,
S ~aryloxy, arylalkoxy, heteroaryl, heteroarylalkyl,
heteroarylalkoxy, cycloalkyl, cycloalkylalkyl or
cycloalkylalkoxy;
R2, R3, R4 are independently hydrogen, halo, alkyl,
haloalkyl, alkenyl, alkoxy, aryloxy, aryl, arylalkyl,
alkylmercapto, arylmercapto, cycloalkyl, cycloalkylalkyl,
heteroaryl, heteroarylalkyl, hydroxy or haloalkyl;
R5 is alkyl of at least 2 carbons, alkenyl, alkynyl;
aryl, heteroaryl, arylalkyl, heteroarylalkyl, cycloalkyl,
cycloalkylalkyl, polycycloalkyl, polycycloalkylalkyl,
1S cycloalkenyl, cycloalkenylalkyl, polycycloalkenyl,
polycycloalkenylalkyl, heteroarylcarbonyl, all of the R5
and R6 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,
2S vitro, 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,
3S arylsulfonylamino; with the proviso that when RS is CH3, R6
_ g _
CA 02307889 2000-04-27
WO 99/Z1564 PCT/US98/21750
is not H; and where R5 is phenyl, the phenyl preferably
includes an ortho hydrophobic substituent such as alkyl,
haloalkyl, aryl, aryloxy or arylalkyl;
R6 is hydrogen or C1-C4 alkyl or C1-C4 alkenyl;
S R~ 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 CH2 and R2 ,
R3 and R4 are each H, R1 will be other than 3,3-
diphenylpropyl.
In the formula ITI compounds, where one of R2, R3
and R4 is 6-fluoro, and the others are H, R~ will be other
than 4-O-methoxyphenyl.
U.S. Application Serial No. 472,067, filed June 6,
1995 (file DC2le) discloses compounds of the structure
o
N~N_ R~
X '
R4
or
or
or
R2 O R~
R3. I_
,\
X '
R4
R5, o
fJ-~N- R~
R6
R~
Rs, D N
Rs J
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or
10
o
~3. I. w ~N.~ _
/ i Y.N J
R4
O O
where O is - C- or - s- ;
O
X is: CHRs, -C- ~ -CH- CH- or
O Rs Rto R9 Rt°
R8, R9 and R1~ are independently hydrogen, alkyl, alkenyl,
alkynyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl,
cycloalkyl, or cycloalkylalkyl;
x 3s -(C8Z)m- or -c-
0
wherein m is 2 or 3;
R1 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
l, 2, 3 or 4 groups selected from halo, haloalkyl, alkyl,
alkenyl, alkoxy, aryloxy, aryl, arylalkyl, alkylmercapto,
arylmercapto, cycloalkyl, cycloalkylalkyl, heteroaryl;
fluorenyl, heteroarylalkyl, hydroxy or oxo;
or R1 is a fluorenyl-type group of the structure
~~/Rts ~,--.,/Rts t~ Rts
i
i , i
tt -Rtt-Zt
_ Rtt_ t
'Z or Z
or Z
Rt2 ~ ~ Rt~Z2 ~ ~ Rt2 Z2 Het
R13 V~ R14 Rt3 v ' Rt4 13 Rt4
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is Ris
Het 1
- R11-Z1
or Ri~Z Z
Het 2
R13 R14
Or
R1 is an indenyl-type group of the structure
R13
R14 13\ 14
R \n/ R
11_ 1
- R Z or , R11- Z1 \ / or
12 2 / Rl6a / 16a
R - ZRisa (CH2~ R12- Z2 R
Rl5a
E
(a - 2,3 or 4)
R13
R13 R14 R14
Het ~ He!
_ R11_ Z1 or _ Rii_ Zi ;
R12_ Z2 ~ Rl6a R12_ Z2 / Rl6a
Risa (CH2)a
G ~ Rlsa
Z1 and Z2 are the same or different and are
independently a bond, O, S,
H
- NH- C- ~ - C- or - C- ,
O ~ ~0~2 ~ O alkyl O ~ O OH
with the proviso that with respect to ~, at least one of Z1
and Z2 will be other than a bond; R11 is a bond, alkylene,
alkenylene or alkynylene of up to 10 carbon~atoms; azylene
or mixed arylene-alkylene; R12 is hydrogen, alkyl,
alkenyl, aryl, haloalkyl, trihaloalkyl, trihaloalkylalkyl,
heteroaryl, heteroarylalkyl, arylalkyl, arylalkenyl, cyclo-
alkyl, aryloxy, alkoxy, arylalkoxy or cycloalkylalkyl, with
the provisos that
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(1) when R12 is H, aryloxy, alkoxy or arylalkoxy,
-NH-C- , -N-C-. -C-
then Z2 is o attryt o ~ o or a bond and -
(2) when Z2 is a bond, R12 cannot be heteroaryl or
heteroarylalkyl;
Z is bond, 0, S, N-alkyl, N-aryl, or alkylene or
alkenylene from 1 to 5 carbon atoms; R13, R14~ R15~ ~d R16
are independently hydrogen, alkyl, halo, haloalkyl, aryl,
cycloalkyl, cycloheteroalkyl, alkenyl, alkynyl, hydroxy,
alkoxy, nitro, amino, thio, alkylsulfonyl, arylsulfonyl,
alkylthio, arylthio, aminocarbonyl, alkylcarbonyloxy,
arylcarbonylamino, alkylcarbonylamino, arylalkyl,
heteroazyl, heteroarylalkyl or aryloxy;
Rl5a ~d Rl6a are independently hydrogen, alkyl,
halo, haloalkyl, aryl, cycloalkyl, cycloheteroalkyl,
alkenyl, alkynyl, alkoxy, alkylsulfonyl, arylsulfonyl,
alkylthio, arylthio, aminocarbonyl, alkylcarbonyloxy,
arylcarbonylamino, alkylcarbonylamino, arylalkyl,
heteroaryl, heteroarylalkyl, or axyloxy;
or R1 is a group of the structure
R19
-(CHZ)D-<
R18
wherein p is 1 to 8 and R1~ and R18 are each independently
H, alkyl, alkenyl, aryl, arylalkyl, heteroaryl,
heteroarylalkyl, cycloalkyl or cycloalkylalkyl at least one
of R1~ and R18 being other than H;
or R1 is a graup of the structure
R2o
- Rts
R2~
wherein R19 is aryl or heteroaryl;
R2~ is aryl or heteroaryl;
R21 is H, alkyl, aryl, alkylaryl, arylalkyl,
aryloxy, arylalkoxy, heteroaryl, heteroarylalkyl,
heteroarylalkoxy, cycloalkyl, cycloalkylalkyl or
cycloalkylalkoxy;
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R2, R3, R4 are independently hydrogen, halo, alkyl,
alkenyl, alkoxy, aryloxy, aryl, arylalkyl, alkylmercapto, _
arylmercapto, cycloalkyl, cycloalkylalkyl, heteroaryl,
heteroarylalkyl, hydroxy or haloalkyl;
R5 is independently alkyl, alkenyl, alkynyl, aryl,
alkoxy, aryloxy, arylalkoxy, heteroaryl, arylalkyl,
heteroarylalkyl, cycloalkyl, cycloalkylalkyl,
polycycloalkyl, polycycloalkylalkyl, cycloalkenyl,
cycloheteroalkyl, heteroaryloxy, cycloalkenylalkyl,
polycycloalkenyl, polycycloalkenylalkyl,
heteroarylcarbonyl, amino, alkylamino, 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, cyclohetero-
alkylalkyl, 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, axylcarbonyloxy,
alkylcarbonylamino, arylcarbonylamino, arylsulfinyl,
arylsulfinylalkyl, arylsulfonyl, alkylsulfonyl,
arylsulfonylamino, heteroarylcarbonylamino,
heteroarylsulfinyl, heteroarylthio, heteroarylsulfonyl,
alkylsulfinyl;
R6 is hydrogen or C1-C4 alkyl or C1-C4 alkenyl; all
optionally substituted with 2, 2, 3 or 4 groups which may
independently be any of the substituents listed in the
definition of RS set out above;
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R~ is alkyl, aryl or arylalkyl wherein alkyl by
itself or as part of arylalkyl is optionally substituted
o
with oxo
f ~ f ~ f
Het Het 1 and Het 2
are the same or different and are independently selected
from heteroaryl containing 5- or 6-ring members; and
O
N-oxides thereof; and
pharmaceutically acceptable salts thereof; with
the provisos that where in the first fornnula X is CH2, and
R2, R3 and R4 are each H, then R1 will be other than 3,3-
diphenylpropyl, and in the fifth formula, where one of R2,
R3 and R~ is 6-fluoro, and the others are H, R7 will be
other than 4-(2-methoxyphenyl).
U.S. Provisional application 60/017,224 filed May 5,
1996, discloses a compound which has the structure
o ~o)Q oa
3 ~ 1 2 ~~ 1 R2 ~ L1
RsiWA $iWRl R wLz-$~BiLwRl rya a~ ~Ra
Rx
or or
I ~ is
including pharmaceutically acceptable salts thereof, N-
oxides thereof,
wherein q is 0, 1 or 2;
A is ( 1 ) a bond;
( 2 ) -O- ; or
-N-
(3) R
where R5 is H or lower alkyl, or R5 together with R2 forms
a carbocyclic or heterocyclic ring system containing 4 to 8
members in the ring;
B is a fluorenyl-type group of the structure
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.~' w l w
R~ Re, R3 4,
R
or Het I /~ or
R3, X ~ Ra R9' X ~ Ra
R3 Ra,
Het 1 Het 2 Or
Ra, X Ra
B is an indenyl-type group of the structure
R3
R3,
R3 3.
~,~/ R
or
S R3b ~ ~ or
3 (CH~
R ~~ Rab
S
(a = 2,3 or 4) Raa
R3
R3,
R3 R3~ Het
Het ) f ;
or
R3b
R3b
S Ras (CHy)a Raa
RX is H, alkyl or aryl;
R1 is alkyl, alkenyl, alkynyl, alkoxyl, (alkyl or
aryl)3Si (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, hetero-
arylsulfonyl, -PO(R13) (R14) , (where R13 and R14 are
independently alkyl, aryl, alkoxy, aryloxy, heteroaryl,
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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 L1 (or L2 in the case of R2) at the 2-
position); 1,3-dioxane or 1,3-dioxolane connected to L1 (or
L2 in the case of R2) at the 4-position; the Rl group may
optionally be substituted with 1, 2, 3 or 4 substituents,
which can be any of the R3 or R1 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,
R2o O
R2~
N-
J
R~
inhere d i s : CHR23 ~ - ~c- -CS - c$- or -c= ~ -
~ ' gsa gas RaaRSs
R23~ R24 ~d R25 are independently hydrogen, alkyl, alkenyl,
alkynyl, aryl, arylalkyl, heteroaryl, heteroazylalkyl,
cycloalkyl, or cycloalkylalkyl;
R20~ R21~ R22 are independently hydrogen, halo,
alkyl, alkenyl, alkoxy, aryloxy, aryl, arylalkyl,
alkylmercapto, arylmercapto, cycloalkyl, cycloalkylalkyl,
heteroaryl, heteroarylalkyl, hydroxy or haloalkyl; and
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these substituents may either be directly attached to Rl,
or attached via an alkylene at an open position;
R2 is independently any of the groups set out for
R1, H, polyhaloalkyl, or cycloheteroalkyl, and may be
optionally substituted with one to four of any of the
groups defined for R3 or substituents defined for R1;
L1 is a linking group containing from 1 to 10
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;
L2 may be the same or different from L1 and may
independently be any of the L1 groups set out above or a
singe bond;
R3, R3', R4 and R4' may be the same or different and
are independently selected from H, halogen, CF3, haloalkyl,
hydroxy, alkoxy, alkyl, aryl, alkenyl, aikenyloxy, alkynyl,
alkynyloxy, alkanoyl, vitro, 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 l, 2 or
3 additional rings fused to Ar;
R3a and R3b are the same or different and are
independently any of the R3 groups except hydroxy, vitro,
amino or thio;
f ~ f ~ f
lief Het t and Het 2
are the same or different and independently represent a 5
or 6 membered heteroaryl ring which contains 1, 2, 3 or 4
heteroatoms in the ring which are independently N, S or O;
and including N-oxides;
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X is a bond, or is one of the following groups:
) -s- _
I
(o)n.
) _o-
(3) -
R6
(4) iC\
R~ R8
(5) -C C~
R9 \RioR9'/~10~
(s) -~ i-
R9~ Rio"
or
wherein
~c\
R9 Rio
Y is O, N-R6 or S;
n' is 0, 1 or 2;
R6 is H, lower alkyl, aryl, -C(O)-R11 or
-C (O) -O-R11;
R~ and R8 are the same or different and are
independently H, alkyl, aryl, halogen, -O-R12, or
R~ and R8 together can be oxygen to form a ketone;
R9, R1~, R9' and Rl~' are the same or different and
are independently H, lower alkyl, aryl or -O-R11
R9~ and Rlo" are the same or different and are
independently H, lower alkyl, aryl, halogen or
-O-R11;
Rll is alky or aryl;
R12 is H, alkyl or aryl;
with the following provisos for compound of the structure
0
LZ ~ Li
Rai wA $i ~Ri
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(a) when R1 is unsubstituted alkyl or unsubstituted
arylalkyl, L1 cannot contain amino; _
(b) when Rz is alkyl, L1 cannot contain amino and
oxo in adjacent positions (to form an amido group);
(c) when RZL2A- is H2N-, R1L1 cannot contain amino;
(d) when R1 is cyano, L1 must have more than 2
carbons;
(e) R1L1 must contain at least 3 carbons;
with respect to compounds of formulas I, IA and IB,
where R1 is cycloheteroalkyl, R1 is exclusive of 1-
piperidinyl, 1-pyrrolidinyl, 1-azetidinyl or 1-(2-oxo-
pyrrolidinyl);
with respect to the sulfur containing compounds and
alcohols, R2L2 cannot have an O or N atom directly attached
to S=(0}q or CRX(OH), and for IA, R2L2 cannot be H.
Sum~arv of the Invention
In accordance with the present invention, novel
compounds are provided which are inhibitors of MTP and have
the structure
Ra
R2/!v Li
\O
O
O
R H[~i
RSa
including pharmaceutically acceptable salts thereof, and
stereoisomers and diasteromers thereof wherein
A is (1) a bond;
(2) -O- ; or
-- N-
R ;
where R5 is H or lower alkyl or R5 together with R2 forms a
30 carbocyclic or heterocyclic ring system containing 4 to 8
members in the ring.
0
A~ B~
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B is a fluorenyl-type group of the structure:
R3 / R4. RS / R4.
or Het
R3. X ~ R4 R3' X ~ R4
R3 R4.
or Het'1 Het2 (the above B is also referred to as a
fluorenyl-type ring or moiety); or
R~ X R4
B is an indenyl-type group of the structure
R3
R3.
R3 3~ 3
or ~'w.'~'/ R R R3.
Z ~ R3b ~ ~ or He_l
se (cH~ J or
R I~~ R3b
R9b
(a = 2,3 or 4) Raa S Raa ~~H~a
R3
R3.
Het
f
(the above B is also referred to as
R3b an indenyl-type ring or moiety);
R~
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);
Rsa is H, lower alkyl or aryl;
R1 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, cycloalkylamino, all
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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,
alkenyl-aminocarbonyl, alkylcarbonyloxy, arylcarbonyloxy,
alkylcarbonylamino, arylcarbonylamino, arylsulfinyl,
arylsulfinylalkyl, arylsulfonyl, alkylsulfonyl,
azylsulfonylamino, heteroarylcarbonylamino, hetero-
arylsulfinyl, heteroarylthio, heteroarylsulfonyl,
alkylsulfinyl; or
R1 and R5a can be joined to form a ring of the
structure
R2o O
R2~- I_ \ N
a
J
R~
cohere a is : c»23, - c- -ca._ cg- or -c= c_ :
I I
0 ~ Ra4 Rzs Rz~ Ras
R23, R24 and R25 are independently hydrogen, alkyl, alkenyl,
alkynyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl,
cycloalkyl, or cycloalkylalkyl;
R2o~ R21~ R22 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
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attached to Rl, or attached via an alkylene chain at an
open position.
R2 is hydrogen, alkyl, alkenyl, alkynyl, haloalkyl,
alkoxyl, (alkyl or aryl)3Si (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(R13)(R14), (where R13 and R14 are
independently alkyl, aryl, alkoxy, aryloxy, heteroaryl,
heteroarylalkyl, heteroaryloxy, heteroarylalkoxy,
cycloheteroalkyl, cycloheteroalkylalkyl, cycloheteroalkoxy,
or cycloheteroalkoxyalkyl); or cyano, 1,1-(alkoxyl or
aryloxy)Zalkyl (where the two aryl or alkyl substituents
can be independently defined.
The R2 group may optionally have from one to four
substituents, which include any of the R1 substituents, and
any of the preferred R2 substituents set out below.
Preferred R2 group when L2 and A are each a single
bond include the following: haloalkylamino,(where halo
includes CF3), alkylamino, cycloalkylamino, arylamino,
heteroarylamino, alkoxyamino, aryloxyamino,
heteroaryloxyamino, heterocyclylamino (where the
heterocycle is connected to the carbonyl group via a
nitrogen or carbon atom).
L1 is a linking group containing from 1 to 10
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
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with alkyl or aryl, an oxo group; and may be substituted
with one to five alkyl or halo groups (preferably F). _
L2 may be the same or different from L1 and may
independently be any of the L1 groups set out above or a
S single bond.
R3, R3', R4 and R4' may be the same or different and
are independently selected from H, halogen, CF3, 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;
R3a and R3b are the same or different and are
independently any of the R3 groups;
f ~ f ~ f
Het Het t and Het 2
'
are the same or different and independently represent a 5
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:
(i) -s-
I
(O)a~
(2) _p_
(3) - I-
R6
(4)
R~ R8
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(5) -C ~'
_ g9 'giogg~~~0~ _
(s) ~ i--
g9~ gio~
(7) ~C' Y-
g9 gio
wherein
Y is 0, N-R6 or S;
n' is 0, 1 or 2;
R6 is H, lower alkyl, aryl, -C(O)-R11 or
-C ( O ) -O-R11;
R7 and R8 are the same or different and axe
independently H, alkyl, aryl, halogen, -O-R12, or
R7 and R8 together can be oxygen to form a ketone;
R9, R1~, R9' and R1~' are the same or different and
are independently H, lower alkyl, aryl or -O-R11;
R9" and Rlfl" are the same or different and are
independently H, lower alkyl, aryl, halogen or
-O-R11
R11 is alley or aryl;
R12 is H, alkyl or aryl;
with the proviso that when A is a (1) bond, R2L2
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, t-
octylamine, 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.
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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.
filed Description of the Invention
The following definitions apply to the terms as used
throughout this specification, unless othezwise limited in
specific instances.
The term "MTP" refers to a polypeptide or protein
complex that (1) if obtained from an organism (e. g., cows,
humans, tec.), can be isolated from the microsomal fraction
of homogenized tissue; and (2) 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 , Naturg ~, 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.
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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
20 carbons, more preferably 1 to 12 carbons, in the normal
chain,such as methyl, ethyl, propyl, isopropyl, butyl, t-
butyl, isobutyl, pentyl, hexyl, isohexyl, heptyl, 4,4-
dimethylpentyl, 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 R3 groups, or the R1
substituents set out herein.
Unless othervuise 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 R3 groups, or the R1
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
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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 [3.3.0]-
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, [2.2.1]-
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, cyclo-
alkylalkyl, cycloheteroalkyl, cycloheteroalkylalkyl, aryl,
heteroaryl, arylalkyl, aryloxy, aryloxyalkyl, arylalkoxy,
arylthio, arylazo, heteroarylalkyl, heteroarylalkenyl,
heteroarylheteroaryl, heteroaryloxy, hydroxy, vitro, 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,
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alkylthio, arylthio, heteroarylthio, arylthioalkyl,
alkoxyarylthio, alkylcarbonyl, arylcarbonyl, alkyl-
aminocarbonyl, arylaminocarbonyl, alkoxycarbonyl,
aminocarbonyl, alkylcarbonyloxy, arylcarbonyloxy,
alkylcarbonylamino, arylcarbonylamino, arylsulfinyl,
arylsulfinylalkyl, arylsulfonylamino or arylsulfon-
aminocarbonyl or any of the R3 groups, or the R1
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
2S 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 carbon 1
y group; examples of
acyl groups include alkanoyl, alkenoyl, aroyl, aralkanoyl,
heteroaroyl, cycloalkanoyl, and the like.
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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
20 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, 2-
propenyl, 3-butenyl, 2-butenyl, 4-pentenyl, 3-pentenyl, 2-
hexenyl, 3-hexenyl, 2-heptenyl, 3-heptenyl, 4-heptenyl, 3-
octenyl, 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, alkex~yl, alkynyl, aryl,
arylalkyl, cycloalkyl, amino, hydroxy, heteroaryl,
cycloheteroalkyl, alkanoylamino, alkylamido,
arylcarbonylamino, vitro, cyano, thiol, alkylthio or any of
the R3 groups, or the R1 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
20 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, 3-
butynyl, 2-butynyl, 4-pentynyl, 3-pentynyl, 2-hexynyl, 3-
hexynyl, 2-heptynyl, 3-heptynyl, 4-heptynyl, 3-octynyl, 3-
nonynyl, 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, vitro, cyano, thiol, and/or
alkylthio, or any of the R3 groups, or the R1 substituents
set out herein.
The term "alkylene" as employed herein alone or as
part of another group refers to alkyl groups as defined
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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 teams "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 (CH2)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 R3 groups, or the R1 substituents set out
herein.
Examples of alkylene, alkenylene and alkynylene
include
-CH=~H-CHZ- ! -CgaCg~Cg- ! -C-C-CHZ- !
-- CH2 C- CHZ CHy CHZ C--.
O O
~3
'_ CgZC = CC82 - ~ - Ca Cg - CHZ ,
'-~~s)s- ! -(CHZ)s- ! -(CHz)a- !
~3
-- (C$2 ) 2- ~ - CHZCBa- ! - CHgCH- ! '- Cg2CgCgZ~ !
~3
CH3 C2H5
C8Z- ! -_ ~ CH~C$Z- ! - CH ~ CHZ- !
~3 C2H5 l ~3
~3
F
CH3
-Cg2-C_CHa- f -(C8a)~ ~ --(CH~)a-C-CHI- ,
~3
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Cl ~3 ~3
-c$a-~-Via- ~ -(~a)a- i'H- ~ -cHa-cH- i- s
C83 C83
- csa - ca - cH - cHa , - cHa - ~H- cHa - ~ H- ,
~3 ~3 CHg CHg
OCHg
1CH3
- CH- CHaCHa- y- CH _ CHaCHa- ~ -~ CHaOCBa-
- OCHaCHa- ~ ~ CHaN8C8a- ~ -- NBCHaCHa-
ICH3 -' -- CHaC$a_.
(CHa)3-CFa- ~ -cga-N-cga-
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
def fined herein .
The term "cycloheteroalkyl" as used herein alone or
as part of another group refers to a 5-, 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 (CHZ)p (which is
defined above), such as
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O\ N~O S'
O
N~ O ~ N
' ' J '
O
N ~/ O
s
and the like. The above groups may include 1 to 4
substituents such as alkyl, halo, oxo and/or any of of the
R3 groups, or the R1 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 4-, 5-, 6- or 7-
membered 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
o~ N s.~ o~N~.
,
N O "'i~ O , V ' p
N ~ N ~/
O/ ~ O~ ~ N V
and the like. The above groups may include 1 to 4
substituents such as alkyl, halo, oxo and/or any of of the
R3 groups, or the R1 substituents set out herein. In
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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
N~ S 0
~ ~ / ~ ~ / s
N~~ N
I
U
N
~N N N~ N
N~/O N~/S
~N,H N~N ~ N ~ '
i5
N._ N ~O
% ~ / ~ / ~ ~ ~ ~!~
<>>~ >>~> ~ ~ 'U
N ~ I
and the like.
Ar may be either aryl or heteroaryl as defined
above.
f ~ f ~ f
Het Het t
and Het 2
'
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 1,2-
positions). Examples of such groups include
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O
/ I '~ / I '~ , I '~ ~ '~ _
Nr~~ N~~ .~ ~ I
O
O i
I , I ~ I '~ I
0
/I
I '
y
~.r' .r~' ft' "
p' ~ '~ .r'~ '
N~ ~~ 1 / a ~ O
N'
s~
N 't, a N ~ ~ N,
/ ~/
N
/i'~ ~ /w~,~
N ' I u,~
wherein a is selected from O, S, and NR7a;
R7a is H, lower alkyl, aryl, -C (O) R7b, -C (O) OR7b%
R7b is alkyl or aryl.
5 The heteroazyl groups including the above groups may
optionally include 1 to 4 substituents such as any of the
R3 groups, or the R1 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 (CH2)p chain.
The term "heteroarylalkyl" or "heteroaryl-alkenyl"
as used herein alone or as part of another group refers to
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a heteroaryl group as defined above linked through a C atom
or heteroatom to a -(CH2)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 C1,
preferably F, such as CF3CH2, CF3 or CF3CF2CH2.
Preferred are compounds of formula I wherein A is
NH,
B is
Ra Rs
X
X is a bond, oxygen or sulfur; R3 and R4 are
independently H or F.
Preferred R2 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 R2 substituents
above).
It is preferred that Ll contains 2 to 7 atoms in the
linear chain and L2 is a bond or lower alkylene.
The preferred L2 group is alkylene such as CH2 or a
bond.
The preferred A group is -NH-.
Preferred R1 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 R1C0, n and the
geometry are as indicated below.
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COHHCHzCFs
O
\ ~CH2)n P~~ -
O
/ ~ ~ '~ NHCOR~
Preferred Structures of Cyclic Phosphonates
R1 CO = n geometry R' CO = n geometry
CF3
CsH
I / ~ traps H CO ~ traps
/ CO
\I
CF3
\ /
( / 1 cis / CO ~ traps
/ ~o \
\
~ N 1 traps
/ I CO 1 traps
/ CO
\I
CF3
I\
C
/ 2 traps ~ 1 traps
/ co / co
\I \I
CF3 /
I \ -
1 traps
2 cis ~ N
/ CO / CO
\ I \ I
O
/ OC- 1 traps
\I
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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.
Acylatioa
Ra OH (carbodiimide coupliaQ) Ra O8
RICOaH, R1N=C=NRZ, BOBt
lA. NS NCORl
Rb R5a Rb R5a
OB OH
Ra O8 ACylBtion Ra OCOR1 Ra OH
3 R1COC1, 3 EtgN 1 Solvolysis
18. pg NCOR Me08, NaH NCORl
b R5a R5a ~ R5a
R OH Rb OCORl Rb OH
Ra OH Silylation Ra OSi(CH3)3 1. Acylatioa Ra OH
I(~g)gSiIZNH (R1COC1, Et3N)
1C. pg
~5a ~S8 ~ 2. Msthaaolysis) NCOR
Rb R Rb R5a (M~OH, HCl) R5a
OH OSi(C8g)g Rb O$
As seen in Scheme lA, serinol derivative (~) can be
acylated via carbodiimide coupling directly to 3_.
Alternatively, as seen in Scheme 1B, ~ can be acylated with
acid chlorides to tri-acylated ,~,. Subsequent solvolysis
provides diols 3_. As seen in Scheme 1C, the diols 3_ could
also be prepared from bis-O-trimethylsilylated amine ~,
acylation with acid chlorides and subsequent methanolysis
to provide ~.
It will be appreciated that in the reactions to
follow, unless otherwise indicated, the moiety "B° in the
starting materials, intermediates and final products is set
out as
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I / x I /
for purposes of illustration only.
It will be appreciated that the "B" 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
l w
Ra. ./. ~ Ra, Ra. Re,
R3 ~' ~ ( = R° ~ R3 Het ~ /~ R4 Or
X~ X
R3.
R4.
R3 R4
Het 1 Het
X as well as any of indenyl-type groups
R3
R3.
R3 3.
or ~~~ R'
S R3b ~ ~ or
a (CH~
R SC ~~ _.
(a - 2,3 or 4) Rsa
R3
R3.
R3 R3.
Het
Het ~ f .
I or
R3b ~ R3b
Rsa (CH~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
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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
\
I / X I / .
The compounds of formula I of the invention may be
prepared as shown in Reaction Scheme 2.
Schenne 2
1. TMSBr
2. C1COCOC1
\ ~ \ Ra O8
/ / O
O ILl-.._ p~0alkyl 3' NCORl, amiae base
R5a
A Oalkyl Rb OH
LsRz
\ ~\ ~\ ~\
+ /
/ / O Ra
O L1 II Ra O L1
~~ O A O O
b 1
LZR2 Rb NCORi LZRZ R 8 RS° R
H R5a 7-Cis
Io 7-trees Ip
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 ~, (which may be chiral or achiral) (as formed in
Scheme lA, 1B or 1C) and an amine base such as
triethylamine to give a mixture of the 7-traps and 7-cis
isomers (Io and Ip, respectively) of the invention. The 7-
trans and 7-cis isomers may be separated by conventional
chromatographic techniques.
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Scheme 3
Preparation of Other Acyl Derivatives From 7-trans
Compounds
~ i ~ _i o
II Ra
O~L~ p~ O
LZR2 Rb NCORl.
H R5s
(where R1' is as
I°~ arylmsthyloxy or
(7-traas) heteroarylmathyloxy)
Hydrogeaolysis
1. 8a, Pd-C/EtOH
2. RICOaH, R1N=C=NRZ, HOBt
Io' Io
~ O
II Ra
O Ll- P~
O
Z Rb NCORl
g RSa
Io
7-traa8
As seen in Scheme 3, the 7-traps isomer Io'
(R1'=OCH2C6H5) (and the corresponding 7-cis isomer) may be
made to undergo hydrogenolysis to form an amine
intermediate which can be reacted with R1COZH to prepare
other aryl derivatives in accordance with the invention.
The starting material Im may be prepared according
to the following Reaction Schemes 4 and 4A.
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Reaction Scheme 4
X
X , 1 ) base
2) Hal-L~-Y
O L~-Y
O Y=Hal
A or O-PG ~ Ih (Y=Hal)
L2R2 XIVa (Y=OPG)
VIII LZR2
Y=OPG
\ X ~ \ Deprotection
F- (for example,
i i n-Bu4NF)
O \Li-OH
A
L2R2 =g
Halide
Formation
X
~ ,
0
\L~-Hal
A
L2R2 Ih
where PG is as oxygea protoctiag group, such
as t-Bu(C83)ZSi or t8u(Ph)gSi-
X
Arbuzov I ~ I ~ ~ Oalkyl
Reaction O
Ih ~ L1~ Oalkyl
Excess (AlkylO)2P(OG1~)
im
-20°C to 180°C L2R2
(Q~ is alkyl, triorganosiiyl (such as trimethylsilyl
or t-butyldimethyisilyl), H, the latter in the
presence of base such as butyilithium, sodium
hydride, or sodium bis-(trimethylsilylamide)
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Reaction Scheme 4A
(A~ternative Mgthod fQr Makina Ih)
1 ) base /
2) Ha1-L~-Y ~
~r o _Lm x
3)C1COCOCU DMF N-R5
0 08 4) (R21-2)RSNH izRs
II Ih
Protected alcohol XIVa can be converted into a wide
variety of functional groups through the intermediary 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 (NaI or KI in acetone or 2-butanone),
or by reaction with triphenylphosphine, I2 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, (alkyl0)3P or (alkyl0)2POSi(alkyl)3 or
(alkyl0)2POH, 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°C to about 180°C.
The starting material VIIIA for compound Im may be
prepared as shown in Reaction Scheme 5.
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Reaction Scheme 5 (Amides)
Preparation of Compounds of Formula I where A is
- N
R5
O
2
COOH R L ~ ~ C
1) amide formation R5
e.g. 1) acid chloride formation ~ , ~ ,
2) (R2L2)RSNH ~X' v
vi=~
A= -N-
~5
R
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 (DID')) to
form the acid chloride IIA
o
CI- C
x
xxar
Acid chloride IIA, without separation from the reaction
mixture, is treated with amine (R2L2)R5NH at a reduced
temperature within the range from about -40°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 (R2L2)R5NH can be carried out via standard literature
procedures.
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Ruction Scheme 6 !Class Esters)
Preparation of Esters VIIIA (A - -0-)
O
H02 H 1) Acid, R2L20H or RZL~-O-C H
2) (COCI)2, R2L20H
3) DCC, HOST,
DMAP, R2L20H
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 H2S04 or p-toluene-sulfonic acid in the presence of an
alcohol such as ally! 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 ((CF3C00)2 or i-BuOCOCl) or
formation of the acylimidazole (carbonyldiimidazole) or
with DCC and HOBT in the presence of DMAP (4-dimethyl-
aminopyridine). These activated intermediates readily form
esters upon treatment with alcohols.
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Ruction Scheme 7 (Amides from Isoc3ranates )
Preparation of Amides VIIIC (A is NH)
O
RzL2HN ~~
\ \
1) base
/ X I / 2) R2L2NC0 [ / I
X
iy v===c
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
Alt~~at~ Scheme for ~om~ound Im2
\ X I \ ~ \ X ~ \
/ / Deesterification
~O OAlkyl O OTMS
t u/ > O 1 u/
O-~L -P TMSBr or TMSI L '-p
A \OAlkyl (Optional tertiary ~A \OTMS
R2L2 amine base) R2L2
Im
Disilyl Ester
Intermediate
Phosphonate Ester Formation
(COCI)2, DMF, CH2CI2
to 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 man~nalian species by administering a
therapeutically effective amount of a compound to decrease
the activity of MTP.
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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:
(1) bovine liver microsomes,
(2) HepG2 cells (human hepatoma cells) or
(3) 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, e~., 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
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(such as mannitol), anti-oxidants (ascorbic acid or sodium
bisulfate) 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 60
(catalog #9385-9), 230-400 mesh under 10-20 psi of nitrogen
pressure.
~ple 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.
N~ CF3 CF3
~~O C /
I\
traps isomer
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Example 2
(Z)-9-[4-[2-Oxo-5-[[[4'-(trifluoromethyl)[1,1'-biphenylJ-2-yl]carbonylJaminoJ-
1,3,2-dioxaphosphorinan-2-ylJbutylJ-N-(2,2,2-trifluoroethyl)-9H-fluorene-
9-carboxamide, N-oxide.
O
/ N/~ CF3 CF3
H I\
\ _ w--.~ o /
O'P~O
\/ O ~ _. . \
A.
cis isomer
OH OH
H \I
O /
\I
CF3
To a stirred solution of 1.33 g (5.00 mmol) of 4'-
trifluorophenyl-2-benzoic acid (Aldrich Chemical Co.),
0.455 g (5.00 mmol) of serinol (Aldrich Chemical Co.),
0.750 g (5.0 mmol) of HOBt and 0.5 mL (3.6 mmol) of
triethylamine in 10 mL 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 (MgS04) 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.
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B.
CONHCHzCFa
P03Et2
B(1) .
O
-OH
Br
To a solution of 9-fluorenecarboxylic acid (Aldrich)
(50 g, 240 mmol) in THF (1200 mL) at 0°C was added dropwise
a solution of n-butyllithium (2.5M, 221 mL, 530 mmol) in
THF. The yellow reaction was stirred at O~C for 1 h, then
1,4-dibromobutane (31.3 mL, 260 mmol) was added dropwise
over 30 min. The reaction was stirred at O~C for 30 min,
then the reaction was warmed to RT fox 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 (1N,
500 mL), then extracted with dichloromethane (3 x 750 mL).
The combined organic layers were dried over MgSO~.
Evaporation gave title compound (71 g, 85~) as a white
solid.
B(2) .
Hn CF3
Br
To a solution of Part B(1) acid (60 g, 173 mmol) and
DMF (100 11.L) in CH2C12 (600 mL) under argon at 0'C was
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added oxalyl chloride (104 mL, 2. OM in CH2C12, 208 nunol)
dropwise. The reaction was stirred at 0'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 CH2C12 (500 mL) at O~C
under argon was added triethylamine (73 mL, 521 mmol)
followed by dropwise addition of a solution of the crude
acid chloride in CH2C12 (15 mL). The reaction was stirred
at 0'C for 1 h, diluted with CH2C12 (500 mL), and washed
with water (2 x 300 mL), 1N HC1 (2 x 300 mL), saturated
NaHC03 (2 x 300 mL), and brine (2 x 300 mL), then dried
over MgS04. 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 CH2C12 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) .
cOtyHGH2Cfa
P09Et2
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 (M+H).
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Anal. Cald'd for C24H2gN04PF3+0.13 mol H20:
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. (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 (Exam~~le 1 )
D. (Z)-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 (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 N,L 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 (MgSO,) 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°C, 205 mg,
27~ yield.
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MICROANALYSIS Calculated for C37 H33 FsN20sP + 0.55 H20:
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~.
1H NMR, i3C NMR, IR and mass spectrometry were consistent
for the indicated compound.
IO
MICROANALYSIS Calculated for C37 Hs3 F6N205P + 064 H20:
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.
Examble 3
traps-9-[4-[5-(Benzoytamino)-2-oxo-1,3,2-dioxaphosphorinan-2-yl]butyl]-
N-(2,2,2-trifluoroethyl)-9H-fluorene-9-carboxamide.
O
/ N CF3
H
O
/ Oy,,
'~- N
HH /
traps isomer
A. N-(1,3-Dibenzoxyloxy-2-propyl)benzamide
OCOCsHS
H
O OCOCsNs
To a stirred slurry of 1.82 g (20.0 mmol) of serinol and
9.2 mL (66 mmol} of triethylamine in 20 mL of
dichloromethane at -5°C under argon, was added dropwise,
7.0 mL (61 mmol) of benzoyl chloride (Aldrich) over 30 min.
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After 2 h, the reaction was diluted with ethyl acetate and
washed three times with 1 ~ hydrochloric acid, once with
brine, twice with saturated sodium bicarbonate solution,
dried (MgSO,) and evaporated. The resulting white solid
was recrystallized from ethyl acetate/hexane to provide
title compound, mp 96-98°C, 5.62 g, 70~ yield.
B. N-(1,3-Dihy~lroxv-2-propyl)benza~ide
OH
H
O OH
To a stirred solution of 160 mg (4.0 mmol) of 60~
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 1.5
mL of 4 I~ 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-1,3,2-
dioxaphosphorinan-2-yl]butyl]-N-(2,2,2-
trifluoroethyl)-9H-fluorene-9-carboxamid.e
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 N,L of DMF. After 2 h, the reaction mixture
was evaporated and redissolved in 3 mL of dichloromethane.
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To this solution at room temperature under argon, was added
283 mg (1.45 mmol) 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 (MgSOq) and evaporated. Purification
by flash chromatography on silica gel (5 x 15 cm column,
ethyl acetate) provided title compound, white solid, mp
166-168°C, 245 mg, 29~ yield.
MICROANALYSIS Calculated for C3oH3oF3N205P + 0.5 H20:
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.
~ lie 4
traps-9-[5-[2-Oxo-5-[[[4'-(trifluoromethyl)[1,1'-biphenyl]-2-
yl]carbonyl]amino]-
1,3,2-dioxaphosphorinan-2-yl]pentyl]-N-(2,2,2-trifluoroethyl)-9H-fluorene-9-
carboxamide.
_ O CF3
/ N/~ CF3
H I \
\ V~ ~,~0 O /
/ O~N I \
traps isomer
~ In a 5
cis-9-[5-[2-Oxo-5-{[[4'-(trifluoromethyl)[1,1'-biphenyl]-2-yl]carbonyl]amino]-
1,3,2-dioxaphosphorinan-2-yl]pentyl]-N-(2,2,2-trifluoroethyl)-9H-fluorene-
9-carboxamide.
O
/ N/~ CF3 CF3
I\
\ /
O ~' O O
/ O~ \
N I
cis isomer
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A. 2-Amino-1,3-bis(trimethylsilyloxy)propane
ost(cH3b
_ ~ NH2
OSi(CH3b
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 by 44-47°C to give title compound as a colorless
liquid, 4.05 g, 86~.
B. N-[(1,3-Dihydroxy)-2-propyl]-2-(4-tri-
fluoromethylphenyl)benzamide
OH OH
H
O /
CFa
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 0°C 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 50
mL of methanol and treated with 0.5 mL of 4 N_t 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~k yield.
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C.
CONHCHzCF3 _
w _
~ ~ ~ P03Eiz
C(1) .
O
OH
Br
To a solution of 9-fluorenecarboxylic acid (Aldrich)
(10 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°C for 1 h, then
1,5-dibromobutane (16.8 mL, 124 mmol) was added dropwise
over 30 min. The reaction was stirred at 0°C 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 (1N, 100
mL), then extracted with dichloromethane (3 x 150 mL). The
combined organic layers were dried over MgS04. Evaporation
gave title compound as a white solid.
C(2) .
CF3
Br
To a solution of all of Part C(1) acid and DMF (20
u.L) in CHZC12 (120 mL) under argon at 0'C was added oxalyl
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chloride (24 mL, 2.0 M in CH2C12, 48 mmol) dropwise. The
reaction was stirred at 0'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 CHZC12 (125 mL) at 0°C under argon was
added triethylamine (16 mL, 115 mmol) followed by dropwise
addition of a solution of the crude acid chloride in CH2C12
(15 mL). The reaction was stirred at 0'C for 1 h, diluted
with CH2C12 (120 mL), and washed with water (2 x 100 mL),
1N HC1 (2 x 100 mL), saturated NaHC03 (2 x 100 mL), and
brine (2 x 100 mL), then dried over MgS04. Evaporation
gave 17.6 g of an oil which was purified by flash
chromatography on silica gel (500 g). The crude product
was loaded in a mixture of CHZC12 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 (14.7
g, 72~) as a white solid ( m.p. 92-96°C).
C(3) .
CF3
Ht~--~ O ~-CHa
O ~~ ~CH3
O
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°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°C) .
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D. traps-9-[5-[2-Oxo-5-[[[4'-(trifluoro-
methyl)[l,1'-biphenyl]-2-yl]carbonyl]amino]-
1,3,2-dioxaphosphorinan-2-yl]pentyl]-N-(2,2,2-
trifluoroethyl)-9H-fluorene-9-carboxamide
SExamDle 4)
E. cis-9-(5-[2-Oxo-5-([[4'-(trifluoromethyl)-
[1,1'-biphenyl]-2-yl]carbonyl]amino]-1,3,2-
dioxaphosphorinan-2-yl]pentyl]-N-(2,2,2-
trifluoroethyl)-9H-fluorene-9-carboxamide
~ .xa le 5 )
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 ~1.L 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°C, 805 mg, 34~ yield.
MICROANALYSIS Calculated for C38H35F6N205P + 0.38 H20:
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.
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The more polar fraction was designated Part E
(Example 5), compound, white foam, mp 110-114°C, 650 mg,
27~.
MICROANALYSIS Calculated for C3gH35F6N205P + 0.38 H20:
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.
~xa~le 6
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, phenylmethyl ester.
O
N/.. CF3
H
0
i o
N
HH /
traps isomer
A. N-Benzyloxycarbonyl-1,3-dihydroxy-2-
aminopropane
OH
H
O~ OH
O
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
10 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 ~ hydrogen chloride in dioxane was added
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and the solution heated to 40°C for 30 min. Evaporation
and trituration with hexane/ether gave title compound as a
white solid, mp 104-106°C, 880 mg, 92~ yield.
MS (electrospray, + ions) m/z 226.
B. traps-[2-Oxo-2-[4-[9-[[(2,2,2-trifluoro-
ethyl)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 ~t.L of DMF. After 2 h, the reaction
mixture was evaporated and redissolved in 5 mL of dichloro-
methane. To this solution at room temperature under argon,
30
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 (MgSO,) 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.
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~~'~.1 1~
traps-1-(Phenylmethyl)-N-[2-[4-[9-[[(2,2,2-trifluoroethyl)amino)carbonylJ-9H-
fluoren-9-yl]butyl]-1,3,2-dioxaphosphorinan-5-yl]-2-piperidinecarboxamide, -
hydrochloride.
O
/ ~ CFa /
H ~O O \
V-- ~.
/ O I H
H
traps isomer
A. N-Benzylpipecolic acid hydrochloride
C02H
~HCI
A mixture of ethyl pipecolinate hydrochloride (10 g,
52 mmol), benzylbromide (7 mL, 57 mmol) and potassium
carbonate (15 g, 114 mmol) in DNIF' (80 mL) was stirred at
room temperature overnight. The solvent was removed in
v cuo. 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
Na2S04, 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. traps-1-(Phenylmethyl)-N-[2-[4-[9-
[[(2,2,2-trifluoroethyl)amino]carbonyl]-9H-
fluoren-9-yl]butyl]-1,3,2-dioxaphosphorinan-5-
vll-2-uineridinecarboxamide. hvdrochloride
An argon-purged slurry of 51.1 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
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argon and filtered through a 0.45 ~ 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
~tL (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 (MgS04)
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 ~! hydrogen
chloride in dioxane and evaporated to provide title
compound as a white solid, mp 125-128°C, 365 mg, 61~ yield.
MICROANALYSIS Calculated for C36HaiF3NsOsP+HCl+1 dioxane+0.33
H20:
C, 59.01; H, 6.27; N, 5.16; C1, 4.35; F, 7.00
Found: C, 59.00; H, 6.25; N, 5.11; C1, 4.14; F, 6.96.
MS (electrospray, + ions) m/e 684.
E~c~mple 8
traps-9-[4-[2-Oxo-5-[[2-(2-pyridinyl)benzoyl]amino]-7 ,3,2-dioxaphosphorinan
2-yi]butyl]-N-(2,2,2-trifluoroethyl)-9H-fluorene-9-carboxamide, hydrochloride.
O
/ N CF3
i\
\ ~'~--\ ~~~° o
/ ~ N \
H
traps isomer
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A. 2-(2-Pyridyl)benzoic acid
O ~N
HO
A(1). 2-Methyl -1-(2-pyridyl)benzene
I ~1
iN
/ I
To a degassed solution of 2-bromopyridine (1.9 mL,
20 mmol) in ethylene glycol dimethyl ether (60 mL) under
argon, tetrakis(triphenylphosphane) palladium (o) (700 mg,
0.6 mmol) was added. After stirring for 10 min, 2-
methylphenyl boronic acid (2.9 g, 22 mmol) was added,
followed by sodium bicarbonate solution (5.04 g, 60 mmol in
60 mL water). The mixture was heated to reflux (-- 85°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 (Na2S04), and the solvent was evaporated to give
a black oil. This oil was distilled at cl Torr at ~95°C to
give title compound (2.75 g, 82~ yield) as a clear oil.
A(2) .
I ~1
~N
COOH
A solution of Part A(1) compound (850 mg, 5.0 mmol)
and potassium permanganate (1.9 g, 12.0 mmol) in water (25
mL) was heated to reflux and stirred for 1 hour. The hot
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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. traps-9-[4-[2-Oxo-5-[[2-(2-pyridinyl)-
benzoyl]amino]-1,3,2-dioxaphosphorinan-2-
yl]butyl]-N-(2,2,2-trifluoroethyl)-9H-
fluor dye-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 ].~, 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 N.L (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 (Na2S04) 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 ~! hydrogen chloride in dioxane and
evaporated to provide title compound as a white solid, mp
132-136°C, 524 mg, 81~ yield.
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MICROANALYSIS Calculated for C35H33F3N305P + HCl + dioxane +
0.25 H20: _-
C, 59.09; H, 5.40; N, 5.30; C1, 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.
Example 9
traps-9-[4-[5-[[2-(2-Benzothiazolyl)benzoyl]amino]-2-oxo-1,3,2-
dioxaphosphorinan-2-yl]butyl]-N-(2,2,2-trifluoroethyl)-9H-fluorene-
9-carboxamide.
0
/ N CFa
H
~N
~, S
P O
O
H
traps isomer
A.
H
O
S
Part A compound is a commercial compound supplied by
Maybridge Chemical Company.
B. traps-9-[4-[5-[[2-(2-Benzothiazolyl)
benzoyl]amino]-2-oxo-1,3,2-dioxaphos-
phorinan-2-yl]butyl]-N-(2,2,2-trifluoro-
e~~y~,-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 ~. nylon filter.
Evaporation and then re-evaporation from toluene gave a
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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 ~,L (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 (Na2S04) 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°C, 269 mg, 62~ yield.
MICROANALYSIS Calculated for C37H33F3N305PS:
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 10
traps-9-[4-[5-[[2-(4-Morpholinyl)benzoyl]amino]-2-oxo-1,3,2-dioxaphosphorinan-
2-yl]butyl]-N-(2,2,2-triouoroethyl)-9H-fluorene-9-carboxamide.
O
/ _ N CF3 O
H
\ ~ ~P, O N
J oJ'
H
traps isomer
A. 2-(1-Morpholino)benzoic acid
C~~
O N
HO
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A(1). Methyl 2-(1-morpholino)benzoate
A solution of methyl 2-fluorobenzoate (15.4 g, 100
mmol) in morpholine (44 mL, 500 mmol) was heated at 50°C
for 30 min, and then warmed to 100°C and stirred for 2 h,
then cooled to 50°C and stirred overnight. The reaction
was then heated to reflex for 2.5 h. The excess morphol.ine
was evaporated. The remainder was dissolved in ethyl
acetate and washed successively with H20 (50 mL), saturated
NaHC03 solution (2 x 50 mL), H20 (3 x 50 mL) and brine (50
mL). Drying (MgS04) and evaporation gave a yellow oil.
The crude product was dissolved in dichloromethane/ethyl
acetate/hexane (4:1:4) 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.
A(2). 2-(1-Morpholino)benzoic acid
O CN
HO
Sodium hydroxide (10 g, 250 mmol) was dissolved in
water (75 mL) and added to a solution of Part A(1) 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 H20 (100 mL) and adjusted to
pH 1.5 with 1N HC1. The aqueous layer was extracted with
chloroform (3 x 250 mL). The combined organic layers were
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dried (Na2S04) and evaporated to give title compound (9.76
g, 85~) as a white solid (m. p. 156-157°C).
B. traps-9-[4-[5-[[2-(4-Morpholinyl)-
benzoyl]amino]-2-oxo-1,3,2-dioxaphosphorinan-
2-yl]butyl]-N-(2,2,2-trifluoroethyl)-9H-
f luo~"rene-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 ).~, 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 N,L (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 (Na2S04) 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°C, 265 mg, 66~ yield.
MICROANALYSIS Calculated for C34H37F3N306P+0.15 H20+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.
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dole 11
traps-9-[4-[5-[[2-(2-Benzoxazolyl)benzoyl]amino]-2-oxo-1,3,2-dioxaphosphorinan-
2-yl]butyl]-N-(2,2,2-trifluoroethyl)-9H-fluorene-9-carboxamide. -
O
I / N~ CFA
H
~N
\ ~~ O O
P. O
/ O~ N '"
H
A. 2-(2-Bromophenyl)benzoxazole
Br
O
To a stirred slurry of 5.46 g (50.0 mmol) of 2-
aminophenol (Aldrich) and 10.05 g (50.0 mmol) of 2-
bromobenzoic 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 refiux
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 ~i
hydrochloric acid, once with water and three times with
saturated sodium bicarbonate solution. The organic phase
was dried (MgSO,) 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~.
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B. 2-(2-Benzoxazolyl)benzoic acid
C02N
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 A~, 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 lh. The reaction
was allowed to warm to room temperature in situ. After 16
h, the mixture was diluted with ether and washed once with
50 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 air-
dried to give 610 mg of title compound as a pink solid, mp
>250°C, 100.
C. trans-9-[4-[5-[[2-(Benzoxazolyl)benzoyl]-
amino]-2-oxo-1,3,2-dioxaphosphorinan-2-
yl]butyl]-N-(2,2,2-trifluoroethyl)-9H-
fluorene-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 mmol) of Part B
compound, 45 mg (0.3 mmol) of HOBt, 65 mg (0.30 mmol) of
EDCI and 20 ElL (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
(Na2S04) 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.
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MICROANALYSIS Calculated for C37H3gF3N306P+2 . 3 H20+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 cis-
isomers, in accordance with the invention, the
corresponding trans-isomers are covered as well.
Fs
I i I i I
HN
F3C-~ O O,p~, O
F I ~ I ~ F F3
i ~ O I
HN
F3C-~ O~P\' O
O O~N w
H
- 71 -
O
R2~LsA~B.LsP~
O~NCOR~
R5a
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WO 99/21564 PCT/US98/21750
I\ I\ I
/ / I\
HN /
F3~ O~P~- O
o ~~ I \
O
/
I \ I \ F3
/ / I\
HN O /
F3G-J O\~p~,
O O°TH~2 -H I \
I \ ~ \ I
/ / I\
HN ~ O / CI
F3CrJ O
O O~H I W
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\ \
I/ I/ I\
HN /
F3C~ O~P\ O
O O~~ I \
CF3
\ ~. N
I/ I/ ~ I\
HN /
F3G- J O~P\~ O
O O~.H I \
/
I \ I ~.. F3
/ / I\
HN ~ O /
FsG-/ O F F U P\'
p H I/
\ \
i/ I/ /I
HN " ~ O O
F G-~ O
O ~Pp-~H I \
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WO 99/21564 PCT/US98/21750
I ~ I ~ -
/ /
HN ' ~ O H2SPh
F3C~ O ~ Pp~-N
H I/
I ~ I ~ F3
/ /
I
HN O /
F3G-J O c~-~
O~ p~N ( w
CH3 /
~' I ~
/ /
HN O CH3
F3G-~ O ~ P~ p~
O _ H I /
F3
I
/ /
H /
O~p~ O
O O~N
N H I /
I ~ I ~
/ /
HN O
F3C~ O
O
/
I ~ I ~ F3
/ /
HN ~ O /
F GJ O p'
3 O, O~H I W
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/ /
HN CH3 O
F3Cr-~ O O P~,.
O~-N
CH3 H
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