Note: Descriptions are shown in the official language in which they were submitted.
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DEMANDES OU BREVETS VOLUMINEUX
LA PRESENTS PARTIE DE CETTE DEMANDS OU CE BREVET
COMPREND PLUS D'UN TOME.
CECi EST LE TOME ~-DE i~- .
NOT'i=: Pour les tomes additionels, veuillez contacter le Bureau canadien des
brevets -
JUMBO APPLfCATIONS/PATENTS
THIS SECTION OF THE APPL1CAT10NIPATENT CONTAINS MORE -
THAN ONE VOLUME
~ THIS IS VOLUME ~' OF
NOTE: -For additiona'1 volumes ~ please contact 'the Canadian Patent Offfice
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HETEROCYCLIC COMPOUNDS AND THEIR USE FOR INHIBITING (~-AMYLOiD PEPTIDE
BACKGROUND OF THE INVENTION
Field of the Invention
This invention relates to compounds which inhibit cellular ~3-amvloid
peptide release andlor its synthesis, and, accordingly, have utility in
treating
Alzheimer's disease.
References
'The following publications, patents and patent applie:ations are cited in
this application as superscript numbers:
' Glenner, et al., "Alzheimer's Disease: lnitial Report of the
Purification and Characterization of a Novel Cerebrovascular
Amyloid -Protein", Biochem. Biophys. Res. Commun. , 120:885-
890 ( 1984) .
= Glenner, et al., "Polypeptide Marker for Alzheimer's Disease
and its Use for Diagnosis." , U. S. Patent No. 4, 666, 829 issued
May 19, 1987.
Selkoe, "The Molecular Pathology of Alzheimer's Disease",
Neuron, 6:487-498 ( 1991 ) .
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° Goate, et ai., "Segregation of a Missense Mutation in the
Amyloid Precursor Protein Gene with Familial Alzheimer's
- Disease", Nature, 349:704-706 (1990).
Chattier-Harlan,- et al., "Early-Onset Alzheimer's Disease Caused
by Mutations at Codon 717 of the a-Amyloid Precursor Proteing
Gene", Nature, 353:844-846 (1989).
Murrell, et al . , "A Mutation in the Amyloid Precursor Protein
Associated with Hereditary Alzheimer's Disease" , Science,
254: 97-99 ( 1991 ).
' Mullan, et al . , "A Pathogenic Mutation for Probable Alzheimer's
Disease in the APP Gene at the N-Terminus of /3-Amyloid,
Nature Genet. , 1:345-347 ( 1992) .
Schenk, et al. , "Methods and Compositions for the Detection of
Soluble (3-Amyloid Peptide", International Patent Application
Publication No. WO 94/10569, published 11 May 1994.
Selkoe, "Amyloid Protein and Alzheimer's Disease", Scientific
American, pp. 2-8, November, 1991.
'o Tetrahedron Letters, 34(48), 7685 ( 1993)
~' Losse, et ai., Tetrahedron, 27:1423-1434 (1971)
Citron, et al. , "Mutation of the ~3-Amyloid Precursor Protein in
Familial Alzheimer's Disease Increases /3-Protein Production,
Nature, 360: 672-674 ( 1992) .
'~ Hansen, et al., "Reexamination and Further Development of a
Precise and Rapid Dye Method for Measuring Cell Growth/Cell
Kill", J. Immun. Meth., 119:203-210 (1989).
All of the above publications, patents and patent applications are herein
incorporated by reference in their entirety to the same extent as if each
individual publication, patent or patent application was specifically and
individually indicated to be incorporated by reference in its entirety.
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State of the Art
Alzheimer's Disease (AD) is a degenerative brain disorder characterized
clinically'by progressive loss of memory, cognition, reasoning, judgment and
emotional stability that gradually leads to profound mental deterioration and
ultimately death. AD is a very common cause of progressive mental failure
(dementia) in aged humans and is believed to represent the fourth most common
medical cause of death in the United States. AD has been observed in races
and ethnic groups worldwide and presents a major present and future public
health problem. The disease is currently estimated to affect about two to
three
million individuals in the United States alone. AD is at present incurable. No
treatment that effectively prevents AD or reverses its symptoms and course is
currently known.
The brains of individuals with AD exhibit characteristic lesions termed
senile (or amyloid) plaques, amyloid angiopathy (amyloid deposits in blood
vessels) and neurofibrillary tangles. Large numbers of these lesions,
particularly amyloid plaques and neurofibrillary tangles, are generally found
in
several areas of the human brain important for memory and cognitive function
in patients with AD. Smaller numbers of these lesions in a more restrictive
anatomical distribution are also found in the brains of most aged humans who
do not have clinical AD. Amyloid plaques and amyloid angiopathy also
characterize the brains of individuals with Trisomy 21 (Down's Syndrome) and
Hereditary Cerebral Hemorrhage with Amyloidosis of the Dutch Type
(HCHWA-D). At present, a definitive diagnosis of AD usually requires
observing the aforementioned lesions in the brain tissue of patients who have
died with the disease or, rarely, in small biopsied samples of brain tissue
taken
during an invasive neurosurgical procedure.
The principal chemical constituent of the amyloid plaques and vascular
amyloid deposits (amyloid angiopathy) characteristic of AD and the other
disorders mentioned above is an approximately 4.2 kilodalton (kD) protein of
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about 39-43 amino acids designated the ~3-amyloid peptide (aAP)-or sometimes
A~3, A~3P or ~3/A4. ~3-Amyloid peptide was first purified and a partial amino
acid sequence was provided by Glenner, et al.' The isolation procedure and the
sequence data for the first 28 amino acids are described in U. S . Patent No .
4, 666, 8292.
Molecular biological and protein chemical analyses have shown that the
~3-amyloid peptide is a small fragment of a much larger precursor protein
(APP), that is normally produced by cells in many tissues of various animals,
including humans. Knowledge of the structure of the gene encoding the APP
has demonstrated that ~3-amyloid peptide arises as a peptide fragment that is
cleaved from APP by protease enzyme(s). The precise biochemical mechanism
by which the ~3-amyloid peptide fragment is cleaved from APP and subsequently
deposited as amyloid plaques in the cerebral tissue and in the walls of the
cerebral and meningeal blood vessels is currently unknown.
Several lines of evidence indicate that progressive cerebral deposition of
,Q-amyloid peptide plays a seminal role in the pathogenesis of AD and can
precede cognitive symptoms by years or decades. See, for example, Selkoe3.
The most important line of evidence is the discovery that missense DNA
mutations at amino acid 7I7 of the 770-amino acid isoform of APP can be
found in affected members but not unaffected members of several families with
a genetically determined (familial) form of AD (Goate, et a1.4; Chartier
Harlan,
et a1.5; and Murrell, et a1.6) and is referred to as the Swedish variant. A
double
mutation changing lysine59s-methionine596 to asparagine59s-leucine596 (with
reference to the 695 isoform) found in a Swedish family was reported in 1992
(MuIlan, et al.') . Genetic linkage analyses have demonstrated that these
mutations, as well as certain other mutations in the APP gene, are the
specific
molecular cause of AD in the affected members of such families. In addition, a
mutation at amino acid 693 of the 770-amino acid isoform of APP has been
identified as the cause of the ~3-amyloid peptide deposition disease, HCHWA-D,
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and a change from alanine to glycine at amino acid 692 appears to cause a
phenotype that resembles AD is some patients but HCHWA-D in others. The
discovery of these and other mutations in APP in genetically based cases of AD
prove that alteration of APP and subsequent deposition of its (3-amyloid
peptide
S fragment can cause AD.
Despite the progress which has been made in understanding the
underlying mechanisms of AD and other /3-amyloid peptide related diseases,
there remains a need to develop methods and compositions for treatment of the
disease(s). Ideally, the treatment methods would advantageously be based on
drugs which are capable of inhibiting (3-amyloid peptide release and/or its
synthesis in vivo.
SUMMARY OF THE INVENTION
This invention is directed to the discovery of a class of compounds
which inhibit /3-amyloid peptide release and/or its synthesis and, therefore,
are
useful in the prevention of AD in patients susceptable to AD andlor in the
treatment of patients with AD in order to inhibit further deterioration in
their
condition. The class of compounds having the described properties are defined
by formula i below:
A-B-C
wherein A is selected from the group consisting of:
R2
H
R~ Z J _ N N
(i) n, H
R6 P
where R' is selected from the group consisting of alkyl, alkenyl,
alkynyl, cycloalkyl, cycloalkenyl, substituted alkyl, substituted alkenyl,
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substituted alkynyl, substituted cycloalkyl, substituted cycloalkenyl, aryl,
heteroaryl and heterocyclic; ,
Z is selected from the group consisting of
(a) a group having the formula -CX'X"C(O)- where X' is hydrogen, ,
hydroxy or fluoro; X" is hydrogen, hydroxy or fluoro, or X' and
X" together form an oxo group;
(b) a group having the formula -T-CX'X"C(O)- where T is selected
from the group consisting of oxygen, sulfur and -NR3 where R3
is hydrogen, acyl, alkyl, aryl or heteroaryl group; X' is hydrogen
or fluoro, X" is hydrogen, hydroxy or fluoro, or X' and X"
together form an oxo group; and
(c) a group having the formula -CX'X"-T-C(O)- where T is selected
from the group consisting of oxygen, sulfur and -NR3 where R3
is hydrogen, acyl, alkyl, aryl or heteroaryl group; X' is hydrogen
or fluoro, X" is hydrogen, hydroxy or fluoro, or X' and X"
together form an oxo group;
R= is selected from the group consisting of alkyl) substituted alkyl,
alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl, aryl,
heteroaryl and heterocyclic;
R6 is selected from the group consisting of alkyl) substituted alkyl,
alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl, aryl,
heteroaryl and heterocyclic;
m is an integer equal to 0 or 1; and
p is an integer equal to 0 or 1;
W X
R~iT,\
(u) C
X~ ~~r
where R' is selected from the group consisting of alkyl, alkenyl,
alkynyl, cycloalkyl, cycloalkenyl, substituted alkyl, substituted alkenyl,
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substituted alkynyl) substituted cycloalkyl, substituted cycloalkenyl, aryl,
heteroaryl and heterocyclic;
T' is selected from the group consisting of a bond covalently linking R'
to -CX'X"-, oxygen, sulfur and -NR3 where R3 is hydrogen, acyl) alkyl, aryl or
heteroaryl group;
W and X are independently selected from the group consisting of
-(CR'R')9 , oxygen, sulfur and -NRg where q is an integer equal to one or two,
and each R' and R8 is independently selected from the group consisting of
hydrogen, alkyl, substituted alkyl, cycloalkyl, aryl, heteroaryl, acyl,
acyloxy,
carboxyl, carboxyl esters and heterocyclic and further, when q is 2, an R'
group on each of the carbon atoms can optionally be fused to form an aryl,
heteroaryl) heterocyclic or cycloalkyl group with the ethylene group with the
proviso that when unsaturated, the remaining R' group on each carbon atom
participates in the unsaturat~on;
and with the further proviso that when W is oxygen, then X is not also
oxygen;
X' is hydrogen, hydroxy or fluoro, X" is hydrogen, hydroxy or fluoro,
or X' and X" together form an oxo group;
R~ is selected from the group consisting of hydrogen, alkyl) substituted
alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl,
substituted cycloalkyl, aryl, heteroaryl and heterocyclic; and
N X
(iii)
/ \ W Ra
x, X,~
where R' is selected from the group consisting of alkyl, alkenyl,
alkynyl, cycloalkyl, cycloalkenyl, substituted alkyl, substituted alkenyl,
substituted alkynyl, substituted cycloalkyl, substituted cycloalkenyl, aryl,
heteroaryl and heterocyclic;
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T' is selected from the group consisting of a bond covalently linking R'
to -CX'X"-, oxygen, sulfur and -NR3 where R' is hydrogen, acyl, alkyl, aryl or
heteroaryl group;
W and X are independently selected from the group consisting of
-(CR'R')~ , oxygen, sulfur and -NRa where q is an integer equal to one or two
and each R' and Rg is independently selected from the group consisting of
hydrogen, alkyl, substituted alkyl) cycloalkyl, aryl, heteroaryl, acyl,
acyloxy,
carboxyl, carboxyl esters, and heterocyclic and further, when g is 2, an R'
group on each of the carbon atoms can optionally be fused to form an aryl,
heteroaryl, heterocyclic or cycloalkyl group with the ethylene group with the
proviso that when unsaturated, the remaining R' group on each carbon atom
participates in the unsaturation;
X' is hydrogen, hydroxy or fluoro, X" is hydrogen, hydroxy or fluoro,
or X' and X" together form an oxo group;
R'' is selected from the group consisting of hydrogen, alkyl, substituted
alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl,
substituted cycloalkyl, aryl, heteroaryl and heterocyclic;
B is selected from the group consisting of:
N
(i)
Rs
where RS is selected from the group consisting of alkyl, substituted
alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl,
substituted cycloalkyl, aryl, heteroaryl and heterocyclic;
W X
(ii) N
W and X are independently selected from the group consisting of
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-(CR'R')q , oxygen, sulfur and -NR$ where q is an integer equal to one or two,
and each R' and Rg is independently selected from the group consisting of
hydrogen, 'alkyl, substituted alkyl, cycloalkyl, aryl, heteroaryl) aryl,
acyloxy,
carboxyl, carboxyl esters, and heterocyclic and further, when g is 2, an R'
group on each of the carbon atoms can optionally be fused to form an aryl,
heteroaryl, heterocyclic or cycloalkyl group with the ethylene group with the
proviso that when unsaturated, the remaining R' group on each carbon atom
participates in the unsaturation;
and with the further proviso that when W is oxygen, then X is not also
oxygen;
R4 is selected from the group consisting of hydrogen) alkyl, substituted
alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl,
substituted cycloalkyl, aryl, heteroaryl and heterocyclic;
N X
(iii)
W and X are independently selected from the group consisting of
-(CR'R'}q-, oxygen, sulfur and -NR$ where q is an integer equal to 1 or 2 and
each R' and Rg is independently selected from the group consisting of
hydrogen, alkyl, substituted alkyl, cycloalkyl, aryl, heteroaryl, aryl,
acyloxy,
and heterocyclic and further, when q is 2, an R' group on each of the carbon
atoms can optionally be fused to form an aryl, heteroaryl, heterocyclic or
cycloalkyl group with the ethylene group with the proviso that when
unsaturated, the remaining R' group on each carbon atom participates in the
unsaturation;
R'~ is selected from the group consisting of hydrogen, alkyl, substituted
alkyl, alkenyl, substituted alkenyi, alkynyl, substituted alkynyl, cycloalkyl,
substituted cycloalkyl, aryl, heteroaryl and heterocyclic; and
(iv) when A is either formula (ii) or (iii) as defined above, then B can
also be a covalent bond linking A to C;
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C is selected from the group consisting of: _
(1) -C(O)Y or -C(S)Y
where Y is selected from the group consisting of:
(a) alkyl or cycloaikyl,
(b) substituted alkyl with the proviso that the substitution on said
substituted alkyl do not include a-haloalkyl, a-diazoalkyl, a-OC(O)alkyl, or
cx-OC(O)aryl groups,
(c) alkoxy or thioalkoxy,
(d) substituted alkoxy or substituted thioalkoxy,
(e) hydroxy,
(f7 aryl,
(g) heteroaryl,
(h) heterocyclic,
(i) -NR'R" where R' and R" are independently selected from hydrogen,
alkyl, alkenyl, alkynyl, substituted alkyl, substituted alkenyl, substituted
alkenyl, cycloaIkyl, aryl, heteroaryl, heterocyclic, where one of R' or R" is
hydroxy or alkoxy, and where R' and R" are joined to form a cyclic group
having from 2 to 8 carbon atoms optionally containing 1 to 2 additional
heteroatoms selected from oxygen, sulfur and nitrogen and optionally
substituted with one or more alkyl, alkoxy or carboxylalkyl groups,
(j) -NHS02-R8 where R$ is selected from alkyl, substituted alkyl,
alkenyl, substituted alkenyl, cycloalkyl, aryl, heteroaryl and heterocyclic,
(k) -NR9NR'°R'° where R9 is hydrogen or alkyl, and each
R'° is
independently selected from hydrogen, alkyl, substituted alkyl, alkenyl,
substituted alkenyl, cycloalkyl, aryl, heteroaryl, heterocyclic, and
(1) -ONR9[C(O)O]ZR'° where Z is zero or one, R9 and R'° are as
defined
above;
(ii) _CR"R"Y'
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where each R" is independently selected from the group consisting of
hydrogen, alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, aryl,
' heteroaryl and heterocyclic and Y' is selected from the group consisting of
hydroxyl, alkoxy, amino, thiol, substituted alkoxy, thioalkoxy, substituted
thioalkoxy, -OC(O)R9, -SSR9, and -SSC(O)R9 where R9 is selected from the
group consisting of alkyl, substituted alkyl, cycloalkyl, aryl, heteroaryl and
heterocyclic; and
(iii) C
N
where A, together with -C = N-, forms a heterocyclic group which is optionally
fused to form a bi- or mufti-fused ring system (preferably no more than S
fused
rings) with one or more ring structures .selected from the group consisting of
cycloalkyl, cycloalkenyl, heterocyclic, aryl and heteroaryl group which, in
turn,
each of such ring structures is optionally substituted with 1 to 4
substituents
selected from the group consisting of hydroxyl, halo, alkoxy, substituted
alkoxy, thioalkoxy, substituted thioalkoxy, vitro, cyano, carboxyl, carboxyl
esters, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl,
substituted
alkynyl; aryl, heteroaryl, heterocyclic, -NHC(O)R'°, -NHSOZR'°, -
C{O)NHz)
-C(O)NHR'°, -C(O)NR'°Rlo, -S(O)R'o, _S(p)zR'o, -
S(O)zNHR'° and
-S(O)~NR'°R'° where each Rl° is independently selected
from the group
consisting of alkyl) substituted alkyl, or aryl, amino, N-alkylamino, N,N-
dialkylamino, N-substituted alkylamino, N,N-disubstituted alkylamino, N-
alkenylamino, N,N-dialkenylamino, N-substituted alkenylamino, N,N-
disubstituted alkenylamino, N-cycloalkylamino, N,N-dicycloalkylamino,
N-substituted cycloalkylamino) N,N-disubstituted cycloalkylamino,
N-arylamino, N,N-diarylamino, N-heteroarylamino, N,N-diheteroarylamino,
N-heterocyclic amino, N,N-diheterocyclic amino and mixed N,N-amino groups
comprising a first and second substituent on said amino group which
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substituents are selected from the group consisting of. alkyl) substiCuted
alkyl,
alkenyl, substituted alkenyl, cycloalkyl, substituted cycloalkyl, aryl)
heteroaryl,
and heterocyclic provided that said first and second substituents are not the
same;
with the proviso that when A has structure (i) and B has structure (i),
then C does not have structure (i) or (ii);
with the further provisos that
A. when A has structure (i) with R' being phenyl) Z being -CH~OC(O}-,
RZ being methyl and p being zero, B has structure (iii) with W being -NH-, X
being -CHz-, and R~ being benzyl then C is not -C(O)OCH3;
B. when A has structure (i) with R' being 3,5-difluorophenyl, Z being
-CHzC(O)-, RZ being methyl, and p being zero, B has structure (ii) with W
being > NC(O)OC(CH3)3, X being -CHI-, and R4 being phenyl, then C is not
-C(O)OCH3; and
C. when A has structure (ii) wherein R' is 3,5-difluorophenyl, T' is a
bond linking R' to -CX'X"-, X' and X" are hydrogen, W is sulfur, X is
methylene and R4 is methyl, and B is a covalent bond linking A to C, then C is
not -C(O)OCH3.
In one preferred embodiment, the compounds of formula I are further
characterized by formula II below:
R H O R5 A
Z N
N N C
mH O ~ H
Rs p \ N
wherein R', R', R5, R6) A, Z, m and p are as defined above. ,
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In formula II above, when m is one, Z is preferably -CX'X"C(O)-
where X" is preferably hydrogen, X' is preferably hydrogen or fluoro or X'
and X" form an oxo group.
S In formula II above, preferred R' unsubstituted aryl groups include, for
example, phenyl, 1-naphthyl, 2-naphthyl, and the like.
Preferred R' substituted aryl groups in formula II include, for example,
monosubstituted phenyls (preferably 3 or 5 substituents}; disubstituted
phenyls
(preferably 3,5 substituents); and trisubstituted phenyls (preferably 3,4,5
substituents). Preferably, the substituted phenyl groups do not include more
than 3 substituents.
Examples of substituted R' phenyls preferred in formula II include, for
instance, 2-chlorophenyl, 2-fluorophenyl, 2-bromophenyl, 2-hydroxyphenyl, 2-
nitrophenyl, 2-methylphenyl, 2-methoxyphenyl, 2-phenoxyphenyl, 2-
trifluoromethylphenyl, 4-fluorophenyl, 4-chlorophenyl, 4-bromophenyl, 4-
nitrophenyl, 4-methylphenyl, 4-hydroxyphenyl, 4-methoxyphenyl, 4-
ethoxyphenyl, 4-butoxyphenyl, 4-iso-propylphenyl, 4-phenoxyphenyl, 4-
trifluoromethylphenyl, 4-hydroxymethylphenyl, 3-methoxyphenyl, 3-
hydroxyphenyl, 3-nitrophenyl, 3-fluorophenyl, 3-chlorophenyl, 3-bromophenyl,
3-phenoxyphenyl, 3-thiomethoxyphenyl, 3-methylphenyl, 3-
trifluoromethylphenyl, 2,3-dichlorophenyl, 2,3-difluorophenyl, 2,4-
dichlorophenyl, 2,5-dimethoxyphenyl, 3,4-dichlorophenyl, 3,4-difluorophenyl,
3,4-methylenedioxyphenyl, 3,4-dimethoxyphenyl, 3,5-difluorophenyl, 3,5-
dichlorophenyl, 3,5-di-(trifluoromethyl)phenyl, 3,5-dimethoxyphenyl, 2,4-
dichlorophenyl, 2,4-difluorophenyl, 2,6-difluorophenyl, 3,4,5-trifluorophenyl,
3,4,5-trimethoxyphenyl, 3,4,5-tri-(trifluoromethyl)phenyl, 2,4,6-
trifluorophenyl,
2,4,6-trimethylphenyl, 2,4,6-tri-(trifluoromethyl)phenyl, 2,3,5-
trifluorophenyl,
2,4,5-trifluorophenyl, 2,5-difluorophenyl, 2-fluoro-3-trifluoromethylphenyl,
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4-fluoro-2-trifluoromethylphenyl, 2-fluoro-4-trifluoromethylphenyl, 4-
benzyloxyphenyl, 2-chloro-6-fluorophenyl, 2-fluoro-6-chlorophenyl, 2,3,4,5,6-
pentafluorophenyl, 2,5-dimethylphenyl, 4-phenylphenyl and 2-fluoro-3-
trifluoromethylphenyl.
Preferred R' alkaryl groups in formula II include, by way of example,
benzyl, 2-phenylethyl, 3-phenyl-n-propyl, and the like.
Preferred R' alkyl, substituted alkyl, alkenyl, cycloalkyl and
cycloalkenyl groups in formula II include, by way of example, iso-propyl,
n-propyl, n-butyl, iso-butyl, sec-butyl, t-butyl, -CH~CH =CH2,
-CHZCH =CH(CHZ)4CH3, cyclopropyl, cyclobutyl, cyclohexyl, cyclopentyl,
cyclohex-1-enyl, -CHZ-cyclopropyl, -CHZ-cyclobutyl, -CHZ cyclohexyl,
-CHZ-cyclopentyl, -CH~CHZ-cyclopropyl, -CHZCHZ-cyclobutyl, -CHZCHZ-
cyclohexyl, -CHZCH2 cyclopentyl, and the like.
Preferred R' heteroaryls and substituted heteroaryls in formula II
include, by way of example, pyrid-2-yl, pyrid-3-yl, pyrid-4-yl, fluoropyridyls
(including 5-fluoropyrid-3-yl), chloropyridyls (including 5-chloropyrid-3-yl),
thiophen-2-yl, thiophen-3-yl, benzothiazol-4-yl, 2-phenylbenzoxazol-5-yl,
furan-
2-yl, benzofuran-2-yl, thionaphthen-2-yl, 2-chlorothiophen-5-yl,
3-methylisoxazol-5-yl, 2-(thiophenyl)thiophen-5-yl, 6-methoxythionaphthen-2-
yl,
3-phenyl-1,2,4-thiooxadiazol-5-yl, 2-phenyloxazol-4-yl, and the like.
Preferably, in formula II, RZ is selected from the group consisting of
hydrogen, alkyl, substituted alkyl, cycloalkyl, aryl, heteroaryl and
heterocyclic.
Particularly preferred RZ substituents include, by way of example, methyl,
ethyl, n-propyl, iso-propyl, n-butyl, iso-butyl, sec-butyl, phenyl, benzyl,
cyclohexyl, cyclopentyl, cyclaheptyl, -CHZCHZSCH3, and the like. As noted
below, R', as well as RS and R6 are preferably the side chain of an L-amino
acid.
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Preferred RS and/or R6 substituents in formula II are independently
selected from the group consisting of include, for example, hydrogen, methyl,
ethyl, n-propyl) iso-propyl, n-butyl, iso-butyl) sec-butyl, tent-butyl,
-CH,CH(CH~CH,),, 2-methyl-n-butyl, 6-f7uoro-n-hexyl, phenyl, benzyl,
cyclohexyl, cyclopentyl, cycloheptyl, allyl, iso-but-2-enyl, 3-methylpentyl,
-CHZ- -cyclopropyl, -CHI-cyclohexyl, -CH,CH~-cyclopropyl, -CHZCHZ-
cyclohexyl, -CHI-indol-3-yl, p-(phenyl)phenyl) o-fluorophenyl,
m-fluorophenyl, p-fluorophenyl, m-methaxyphenyl, p-methoxyphenyl,
phenethyl, benzyl, m-hydroxybenzyl, p-hydroxybenzyl, p-nitrobenzyl,
m-trifluoromethylphenyl, _p-(CH3),NCHZCH,CH,O-benzyl,
p-(CH3)3COC(O)CH~O-benzyl, p-(HOOCCH~O)-benzyl, 2-aminopyrid-6-yl, p-
(N-morpholino-CH,CH,O)-benzyl, -CH,CH~C(O)NH,, -CHI-imidazol-4-yl,
-CH,- _ -(3-tetrahydrofuranyl), -CH,-thiophen-2-yl, -CH,( 1-
methyl)cyclopropyl,
-CH,-thiophen-3-yl, thiophen-3-yl, thiophen-2-yl, -CHI-C(O)O-t-butyl,
-CHz-C(CH3)3, -CH~CH(CHZCH3)2, -2-methylcyclopentyl, -cyclohex-2-enyl)
-CH[CH(CH3)z]COOCH3, -CHZCH,N(CH3)~, -CHZC(CH3)=CH2,
-CH,CH =CHCH3 (cis and trans), -CHZ(~H, -CH(OH)CH3, -CH(O-t-butyl)CH3,
-CH,OCH3, -(CHZ)4NH-Boc, -(CHz)4NH~, -CHI-pyridyl (e.g., 2-pyridyl,
3-pyridyl and 4-pyridyl), pyridyl (2-pyridyl, 3-pyridyl and 4-pyridyl}, -CHZ-
naphthyl -(e. g. , 1-naphthyl and 2-naphthyl), -CH,-(N-morpholino),
p- _(N-morpholino-CHZCH,O)-benzyl, benzo[b]thiophen-2-yl,
5-chlorobenzo[b]thiophen-2-yl, 4,5,6,7-tetrahydrobenzo[b]thiophen-2-yl)
benzo[b]thiophen-3-yl, 5-chlorobenzo[b]thiophen-3-yl, benzo[b]thiophen-5-yl,
6- _methoxynaphth-2-yl, -CH,CH,SCH3, thien-2-yl, thien-3-yl, and the like.
/A
In formula II, preferred C \ groups are further
N
characterized by the following structures:
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Y"
A'
s _ _ c\
N
where Y" is a heteroatom selected from oxygen, sulfur and > NRg where R$ is
as defined above and A', together with -Y"-C=N-) forms a heterocyclic group
which is optionally fused to form a bi- or mufti-fused ring system (preferably
no more than 5 fused rings) with one or more ring structures selected from the
group consisting of cycloalkyl, cycloalkenyl, heterocyclic, aryl and
heteroaryl
group which, in turn, each of such ring structures are optionally substituted
with 1 to 4 substituents selected from the group consisting of hydroxyl, halo,
alkoxy, substituted alkoxy, thioalkoxy) substituted thioalkoxy, nitro, cyano,
carboxyl, carboxyl esters, alkyl, substituted alkyl, alkenyl, substituted
alkenyl,
alkynyl, substituted alkynyl, amino, N-alkylamino, N,N-dialkylamino,
N-substituted alkylamino, N-alkyl N-substituted alkylamino, N,N-disubstituted
alkylamino) aryl, heteroaryl, heterocyclic, -NHC(O)R'°, -
NHSOZR'°,
-C(O)NH,, -C(O)NHR'°, -C(O)NR'°R'°, -S(O)R'°, -
S(O)zR'°, -S(O),NHR'° and
-S(O),NR'°R'° where each R'° is independently selected
from the group
consisting of alkyl, substituted alkyl, or aryl.
Preferred heterocyclic structures include, by way of example, 3-methyl-
1,2,4-oxadiazol-5-yl, thiazolin-2-yl, 3-phenyl-1,2,4-oxadiazol-5-yl, 3-(p-
methoxy-benzyl)-1,2,4-oxadiazol-5-yl) and the like.
Preferred compounds of formula II include the following:
(S)-5-[1-N [N (3,s-difluorophenylacetyl)-L-alaninyl]amino]ethyl-3-ethyl-
1,2,4-oxadiazole
(S)-s-[ 1-N-[N-(3 , 5-difluorophenylacetyl)-L-alaninyl] amino-2-
phenylethyl]-3-methyl-1,2,4-oxadiazole
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__ 1~ -_
2-[1-N [N (3,5-difluorophenylacetyl)-L-alaninyl)amino-1-phenyl]methyl-
2-thiazoline
(S)-5-[1-N [N (3,5-difluorophenylacetyl)-L-alaninyl]amino-1-
phenyl]methyl-3-methyl-1,2,4-oxadiazole
(S)-5-[1-N [N (3,5-difluorophenylacetyl)-L-alaninyl]amino-1-
phenyl]methyl-3-phenyl-1,2,4-oxadiazole
(S)-5-[1-N [N (3,5-difluorophenylacetyl)-L-alaninyl]amino-1-
phenyl]methyl-3-(4-methoxyphenylmethyl)-1,2,4-oxadiazole
In another preferred embodiment, the compounds of formula I are
further characterized by formula III and IV below:
R2 W X
H ~ , /C(O)Y
R~ ~Z~ N N R4 III
H
R6 J P
ana
R2 H N X
C(O)Y
R~ ~Z~ N N W R4 IV
Rs
P
wherein R', RZ) R°, R6, W, X, Y, Z, m and p are as defined above.
In formula III and IV above, when m is one, Z is preferably
-CX'X"C(O)- where X" is preferably hydrogen, X' is preferably hydrogen or
fluoro and where X' and X" form an oxo group.
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In formula III and IV above, preferred R' unsubstituted aryl groups
include, for example, phenyl, 1-naphthyl, 2-naphthyl, and the like.
Preferred R' substituted aryl groups in formula III and IV include, for
example, monosubstituted phenyls (preferably 3 or 5 substituents};
disubstituted
phenyls (preferably 3,5 substituents); and trisubstituted phenyls (preferably
3,4,5 substituents). Preferably, the substituted phenyl groups do not include
more than 3 substituents.
Examples of substituted Rl phenyls preferred in formula III and IV
include, for instance, 2-chlorophenyl, 2-fluorophenyl, 2-bromophenyl,
2-hydroxyphenyl, 2-nitrophenyl, 2-methylphenyl, 2-methoxyphenyl,
2-phenoxyphenyl, 2-trifluoromethylphenyl, 4-fluorophenyl, 4-chlorophenyl,
4-bromophenyl, 4-nitrophenyl, 4-methylphenyl, 4-hydroxyphenyl,
4-methoxyphenyl, 4-ethoxyphenyl, 4-butoxyphenyl, 4-iso-propylphenyl,
4-phenoxyphenyl) 4-trifluoromethylphenyl, 4-hydroxymethylphenyl,
3-methoxyphenyl, 3-hydroxyphenyl, 3-nitrophenyl, 3-fluorophenyl,
3-chlorophenyl, 3-bromophenyl, 3-phenoxyphenyl, 3-thiomethoxyphenyl,
3-methylphenyl, 3-trifluoromethylphenyl, 2,3-dichlorophenyl,
2,3-difluorophenyl, 2,4-dichlorophenyl, 2,5-dimethoxyphenyl,
3,4-dichlorophenyl, 3,4-difluorophenyl, 3,4-methylenedioxyphenyl,
3,4-dimethoxyphenyl, 3,5-difluorophenyl, 3,5-dichlorophenyl, 3,5-di-
(trifluoromethyl)phenyl, 3,5-dimethoxyphenyl, 2,4-dichlorophenyl,
2,4-difluorophenyl, 2,6-difluorophenyl, 3,4,5-trifluorophenyl,
3,4,5-trimethoxyphenyl, 3,4,5-tri-(trifluoromethyl)phenyl, 2,4,6-
trifluorophenyl,
2,4,6-trimethylphenyl, 2,4,6-tri-(trifluoromethyl)phenyl, 2,3,5-
trifluorophenyl,
2,4,5-trifluorophenyl, 2,5-difluorophenyl, 2-fluoro-3-trifluoromethylphenyl,
4-fluoro-2-trifluoromethylphenyl, 2-fluoro-4-trifluoromethylphenyl,
4-benzyloxyphenyl, 2-chloro-6-fluorophenyl, 2-fluoro-6-chlorophenyl,
2,3,4,5,6-pentafluorophenyl, 2,5-dimethylphenyl, 4-phenylphenyl and 2-fluoro-
3-trifluoromethylphenyl.
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Preferred R' alkaryl groups in formula III and IV include,- by way of
example, benzyl, 2-phenylethyl, 3-phenyl-n-propyl, and the like.
Preferred R' alkyl, substituted alkyl, alkenyl, cycloalkyl and
' S cycloalkenyl groups in formula III and IV include, by way of example,
iso-propyl, n-propyl, n-butyl, iso-butyl, ,sec-butyl, t-butyl, -CH~CH=CH2,
-CH,CH =CH(CHZ)4CH3, cyclopropyl, cyclobutyl, cyclohexyl, cyclopentyl,
cyclohex-1-enyl, -CHZ cyclopropyl, -CH~-cyclobutyl) -CHZ-cyclohexyl,
-CHI-cyclopentyl, -CH~CHZ-cyclopropyl, -CH~CHZ-cyclobutyl, -CH~CHZ-
cyclohexyl, -CH~CHZ-cyclopentyl, and the like.
Preferred R' heteroaryls and substituted heteroaryls in formula III and
IV include, by way of example, pyrid-2-yl, pyrid-3-yl, pyrid-4-yl,
fluoropyridyls (including 5-fluoropyrid-3-yl), chloropyridyls (including 5-
chloropyrid-3-yl), thiophen-2-yl, thiophen-3-yl, benzothiazol-4-yl,
2=phenylbenzoxazol-5-yl, furan-2-yl, benzofuran-2-yl, thionaphthen-2-yl,
2-chlorothiophen-5-yl, 3-methylisoxazol-5-yl, 2-(thiophenyl)thiophen-5-yl,
6-methoxythionaphthen-2-yl, 3-phenyl-1,2,4-thiooxadiazol-5-yl, 2-phenyloxazol-
4-yl, and the like.
Preferably, in formula III and IV, R' is selected from the group
consisting of hydrogen, alkyl, substituted alkyl, cycloalkyl, aryl, heteroaryl
and
heterocyclic. Particularly preferred RZ substituents include, by way of
example,
methyl, ethyl, n-propyl, iso-propyl, n-butyl, iso-butyl, sec-butyl, phenyl,
benzyl, cyclohexyl, cyclopentyl, cycloheptyl, -CHZCHZSCH3, and the like. As
noted below, R2 (as well as R6) are preferably the side chain of an L-amino
acid.
Preferred R6 substituents in formula III and IV include, for example,
hydrogen, methyl, ethyl, n-propyl, iso-propyl, n-butyl, iso-butyl, sec-butyl,
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tert-butyl, -CHZCH(CHZCH~)2, 2-methyl-n-butyl, 6-fluoro-n-hexyl, phenyl,
benzyl, cyclohexyl, cyclopentyl, cycloheptyl, allyl, iso-but-2-enyl,
3-methylpentyl, -CHZ-cyclopropyl, -CH4-cyclohexyl, -CHzCH2-cyclopropyl,
-CHzCH2-cyclohexyl, -CHZ indol-3-yl, p-(phenyl)phenyl, o-ftuorophenyl,
m-fluorophenyl, p-fluorophenyl, m-methoxyphenyl, p-methoxyphenyl,
phenethyl, benzyl, m-hydroxybenzyl, p-hydroxybenzyl, p-nitrobenzyl,
m-trifluoromethylphenyl, p-{CH3)~NCH~CHZCH,O-benzyl,
p-(CH3)3COC{O)CH,O-benzyl, p-(HOOCCH~O)-benzyl, 2-aminopyrid-6-yl,
p-(N-morpholino-CHZCH,O)-benzyl, -CH~CH~C{O)NHz, -CHZ-imidazol-4-yl,
-CH,-(3-tetrahydrofuranyl), -CH2-thiophen-2-yl, -CHZ(1-methyl)cyclopropyl,
-CH,-thiophen-3-yl, thiophen-3-yi, thiophen-2-yl, -CH~-C(O)O-t-butyl,
-CHI-C(CH3),, -CHZCH(CHZCH3)2, -2-methylcyclopentyl, -cyclohex-2-enyl,
-CH[CH(CH3)2]COOCH3, -CHZCHZN{CH3)Z, -CHZC(CH;)=CHz,
-CHZCH=CHCH3 (cis and trans), -CHZOH, -CH(OH)CH3, -CH(O-t-butyl)CH3,
-CHZOCH3, -(CHZ)~NH-Boc, -(CHZ)QNHZ, -CHI-pyridyl (e.g., 2-pyridyl,
3-pyridyl and 4-pyridyl), pyridyl (2-pyridyl, 3-pyridyl and 4-pyridyl),
-CHZ-naphthyl (e.g., 1-naphthyl and 2-naphthyl), -CHZ-(N-morpholino),
p-(N-morpholino-CHZCH20)-benzyl, benzo[b]thiophen-2-yl,
5-chlorobenzo[b]thiophen-2-yl, 4,5,6,7-tetrahydrobenzo[b]thiophen-2-yl,
benzo[b]thiophen-3-yl, 5-chlorobenzo[b]thiophen-3-yl, benzo[b]thiophen-5-yl,
6-methoxynaphth-2-yl, -CH~CH~SCH3, thien-2-yl) thien-3-yl, and the Like.
Preferably Y in Formula III or IV is hydroxy, alkoxy, substituted alkoxy
and -NR'R" where R' and R" are as defined above. Preferred alkoxy and
substituted alkoxy groups include rnethoxy, ethyoxy, n-propoxy, iso-propoxy,
n-butoxy, iso-butoxy, t-butoxy, neo-pentoxy, benzyloxy, 2-phenylethoxy, 3-
phenyl-n-propoxy, 3-iodo-n-propoxy, 4-bromo-n-butoxy, and the like.
Preferred -NR'R" groups include, by way of example, amino (-NHZ),
-NH(iso-butyl), -NH(sec-butyl), N-methylamino, N,N-dimethylamino,
N-benzylamino, N-morpholino, azetidino, N-thiomorpholino, N-piperidinyl,
N-hexamethyleneimino, N-heptamethylene-imino, N-pyrrolidinyl,
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-NH-methallyl, -NHCH~-{furan-2-yl), -NHCHZ-cyclopropyl, -NH(t-butyl),
-NH(p-methylphenyl), -NHOCH~, -NHCHZ(p-fluorophenyl), -NHCHZCH~OCH3,
-NH-cyclo~exyl, -NHCHzCH2N(CH3)Z, -NHCHZC(CH3)~, -NHCHZ-(pyrid-2-yl),
-NHCHZ-(pyrid-3-yl), -NHCHZ (pyrid-4-yl), N-thiazolindinyl,
-N(CHZCHZCH3)Z, -NHOH, -NH(p-NOZ-~), -NHCHZ(p-NOZ-~),
-NHCHZ(m-NOZ-~), -N(CH~)OCH3> -N(CH3)CHz ~, -NHCHz-(3,5-di-
fluorophenyl), -NHCH,CH~F, -NHCHz(p-CH~O-~), -NHCHZ(m-CH30-~),
-NHCHZ(p-CF3-~), -N(CH3)CH~CH~OCH;, -NHCHZCH2~> -NHCH(CH3)~>
-NHCHZ- _(p-F-~)> -N(CH3)CHZCH,N(CIi3)2, -NHCHZ-(tetrahydrofuran-2-yl),
and the like.
Still another preferred Y group is an alkyl group such as
-CHZCH~CH(CH3)2, and the like.
Preferred heterocyclic structures defined by W and X include, by way of
example, 4,5-dihydrothiazoles, 3,4-dihydro-1,3-isodiazoles, 3,4-dihydro-3-N-t-
butoxy-3-isodiazoles, 4,5-dihydrooxazoles, and the like.
Preferred compounds of formula III and IV include the following:
(S)-2-[1-(3,5-difluorophenylacetamido)ethyl]-4-ethoxycarbonyl-2-
thiazoline
1-ten-butoxycarbonyl-2-[ 1-(N-carbobenzyloxy)aminoethyl]-4-
methoxycarbonyl-4-phenylmethyl-2-imidazoline
1-tent-butoxycarbonyl-2-[ 1-(3 , 5-difluorophenylacetamido)ethyl]-4-
methoxycarbonyl-4-phenylmethyl-2-imidazoline
2-[ 1-(3 , 5-difluorophenylacetamido)ethyl]-4-methoxycarbonyl-4-
phenylmethyl-2-imidazoline
2-[ 1-(3 , S-difluorophenylacetamido)ethyl)-4-methoxycarbonyl-4-phenyl-2-
imidazoline
2-[ 1-(3 , 5-difluorophenylacetamido)-1-phenyl)methyl-4-ethoxycarbonyl-2-
thiazoline
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1-tent-butoxycarbonyl-2-[1-(N carbobenzyloxy)aminoethyl]-4-
methoxycarbonyl-4-phenyl-2-imidazoline
2-[(S)-I-(3,S-dichloroanilino)ethyl]-(S)-4-methoxycarbonyl-2-oxazolidine
(S)-2-[1-(3,5-difluorophenylacetamido)ethyl]-5(R,S)-ethoxycarbonyl-2- .
oxazoline
2-[1-(3,S-difluorophenylacetamido)-1-phenyl]methyl-4-methoxycarbonyl-
2-thiazoline
[ 1-(N carbobenzyloxy)aminoethyl]-4-methoxycarbonyl-4-phenyl-2-
imidazoline
In another preferred embodiment, the compounds of formula I are
further characterized by formula V and VI below:
p Rs
W X
R~~T'~C~ N Y V
/ \ N R4 H
X' X"
and
Rs
N X
R~iT'~C~ N Y Vi
/ \ W \R4 H O
X' X"
where R', R~, R6, T', X', X", W, X, and Y are as defined above.
In formula V and VI above, when m is one, Z is preferably
-CX'X"C(O)- where X" is preferably hydrogen, X' is preferably hydrogen or
fluoro or where X' and X" form an oxo group.
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In formula V and VI above, preferred R' unsubstituted aryl groups
include, for example, phenyl, 1-naphthyl, 2-naphthyl, and the like.
Preferred R' substituted aryl groups in formula V and -VI include, for
example, monosubstituted phenyls (preferably 3 or 5 substituents);
disubstituted
phenyls (preferably 3,5 substituents); and trisubstituted phenyls (preferably
3 , 4 , 5 substituents) . Preferably , the substituted phenyl groups do not
include
more than 3 substituents.
Examples of substituted R' phenyls preferred in formula IV and VI
include, for instance, 2-chlorophenyl, 2-fluorophenyl, 2-bromophenyl,
2-hydroxyphenyl, 2-nitrophenyl, 2-methylphenyl) 2-methoxyphenyl,
2-phenoxyphenyl, 2-trifluoromethylphenyl, 4-fluorophenyl, 4-chlorophenyl,
4=bromophenyl, 4-nitrophenyl, 4-methylphenyl, 4-hydroxyphenyl,
IS 4-methoxyphenyl, 4-ethoxyphenyl, 4-butoxyphenyl, 4-iso-propylphenyl,
4-phenoxyphenyl, 4-trifluoromethylphenyl, 4-hydroxymethylphenyl,
3-methoxyphenyl, 3-hydroxyphenyl, 3-nitrophenyl, 3-fluorophenyl,
3-chlorophenyl, 3-bromophenyl, 3-phenoxyphenyl, 3-thiomethoxyphenyl,
3-methylphenyl, 3-trifluoromethylphenyl, 2,3-dichlorophenyl,
2,3-difluorophenyl, 2,4-dichlorophenyl, 2,5-dimethoxyphenyl,
3,4-dichlorophenyl, 3,4-difluorophenyl, 3,4-methylenedioxyphenyl,
3 ,4-dimethoxyphenyl, 3 , 5-difluorophenyl, 3 , 5-dichlorophenyl, 3 , 5-di-
(trifluoromethyl)phenyl, 3,5-dimethoxyphenyl, 2,4-dichlorophenyl,
2,4-difluorophenyl, 2,6-difluorophenyl, 3,4,5-trifluorophenyl,
3,4,5-trimethoxyphenyl, 3,4,5-tri-(trifluoromethyl)phenyl, 2,4,6-
trifluorophenyl,
2,4,6-trimethylphenyl, 2,4,6-tri-(trifluoromethyl)phenyl, 2,3,5-
trifluorophenyl,
2,4,5-trifluorophenyl, 2,5-difluorophenyl, 2-fluoro-3-trifluoromethylphenyl,
4-fluoro-2-trifluoromethylphenyl, 2-fluoro-4-trifluoromethylphenyl,
4-benzyloxyphenyl, 2-chloro-6-fluorophenyl, 2-fluoro-b-chlorophenyl,
2 , 3 , 4 , 5 , 6-pentafluoropheny 1, 2 , 5 -dimethy lpheny 1, 4-phenylpheny 1
and 2-fluoro-
3-trifluoromethylphenyl .
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Preferred R' alkaryl groups in formula IV and VI include, by way of
example, benzyl, 2-phenylethyl, 3-phenyl-n-propyl, and the Iike.
Preferred R' alkyl, substituted alkyl, alkenyl, cycloalkyl and
cycloalkenyl groups in formula V and VI include, by way of example,
iso-propyl, n-propyl, n-butyl, iso-butyl, sec-butyl, t-butyl, -CHZCH=CHZ,
-CH~CH=CH(CH2)4CH3, cyclopropyl, cyclobutyl, cyclohexyl, cyclopentyl,
cyclohex-1-enyl, -CHZ-cyclopropyl, -CHI-cyclobutyl, -CH~-cyclohexyl,
-CHZ-cyclopentyl, -CH,CH~-cyclopropyl, -CH~CHz-cyclobutyl, -CH2CHz-
cyclohexyl, -CHZCHZ-cyclopentyl, and the like.
Preferred R1 heteroaryls and substituted heteroaryls in formula III and
iV include, by way of example, pyrid-2-yl, pyrid-3-yl, pyrid-4-yl,
fluoropyridyls (including 5-fluoropyrid-3-yl), chloropyridyls (including 5-
chloropyrid-3-yl), thiophen-2-yl, thiophen-3-yl, benzothiazol-4-yl,
2-phenylbenzoxazol-5-yl, furan-2-yl, benzofuran-2-yl, thionaphthen-2-yl,
2-chlorothiophen-5-yl, 3-methylisoxazol-5-yl, 2-(thiophenyi)thiophen-5-yl,
6-methoxythionaphthen-2-yl, 3-phenyl-1,2,4-thiooxadiazol-5-yl, 2-phenyloxazol-
4-yl, and the like.
Preferred R4 substituents in formula III and IV include, for example,
hydrogen, methyl, phenyl, benzyl and the like.
Preferred R6 substituents in formula V and VI include, for example,
hydrogen, methyl, ethyl, n-propyl, iso-propyl, n-butyl, iso-butyl, sec-butyl,
ten-butyl, -CHZCH(CHZCH3)2, 2-methyl-n-butyl, 6-fluoro-n-hexyl, phenyl,
benzyl, cyclohexyl, cyclopentyl, cycloheptyl, allyl, iso-but-2-enyl,
3-methylpentyl, -CHZ-cyclopropyl, -CHI-cyclohexyl, -CH~CH,-cyclopropyl,
-CHZCHZ-cyclohexyl, -CHZ-indol-3-yl, p-(phenyl)phenyl, o-fluorophenyl,
m-fluorophenyl, p-fluorophenyl, m-methoxyphenyl, p-methoxyphenyl,
phenethyl, benzyl, m-hydroxybenzyl, p-hydroxybenzyl, p-nitrobenzyl,
___._m.,.... _ ._._.~___....
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m-trifluoromethylphenyl, p-(CH3)~NCH,CHzCHzO-benzyl, -
p-(CH3)3COC(O)CHzO-benzyl, p-(HOOCCH20)-benzyl, 2-aminopyrid-6-yl,
p-{N-morpholino-CH~CHZO)-benzyl, -CHZCH~C(O)NHZ, -CHZ-imidazol-4-yl,
-CHz-(3-tetrahydrofuranyl), -CHZ-thiophen-2-yl, -CH~(1-methyl)cyclopropyl,
-CHz-thiophen-3-yl, thiophen-3-yl, thiophen-2-yl, -CHZ-C{O)O-t-butyl,
-CHZ-C(CH3)3, -CH,CH(CH.,CH3)Z, -2-methylcyclopentyl, -cyclohex-2-enyl,
-CH[CH{CH3)Z]COOCH3, -CHZCH~N(CH~)2, -CH,C(CH3)=CHZ,
-CH~CH=CHCH3 (cis and trans), -CHZOH, -CH(OH)CH3, -CH(O-t-butyl)CH3,
-CH~OCH3, -(CHz)4NH-Boc, -(CHZ)4NH2, -CHZ-pyridyl (e.g., 2-pyridyl,
3-pyridyl and 4-pyridyl), pyridyl (2-pyridyl, 3-pyridyl and 4-pyridyl),
-CHZ-naphthyl {e.g., 1-naphthyl and 2-naphthyl), -CHZ-(N-morpholino),
p-(N-morpholino-CH~CH20)-benzyl, benzo[b]thiophen-2-yl,
5-chlorobenzo[b]thiophen-2-yl, 4,5,6,7-tetrahydrobenzo[b]thiophen-2-yl,
benzo[b]thiophen-3-yl, 5-chlorobenzo[b]thiophen-3-yl, benzo[b]thiophen-5-yl,
6-methoxynaphth-2-yI, -CH~CHZSCH3, thien-2-yl, thien-3-yl, and the like.
Preferably Y in Formula V or VI is hydroxy, alkoxy, substituted alkoxy
and -NR'R" where R' and R" are as defined above. Preferred alkoxy and
substituted alkoxy groups include methoxy, ethyoxy, n-propoxy, iso-propoxy,
n-butoxy, iso-butoxy, t-butoxy, neo-pentoxy, benzyloxy, 2-phenylethoxy, 3-
phenyl-n-propoxy, 3-iodo-n-propoxy, 4-bromo-n-butoxy, and the like.
Preferred -NR'R" groups include, by way of example, amino (-NHz),
-NH(iso-butyl), -NH(sec-butyl), N-methylamino, N,N-dimethylamino,
N-benzylamino, N-morpholino, azetidino, N-thiomorpholino, N-piperidinyl,
N-hexamethyleneimino, N-heptamethylene-imino, N-pyrrolidinyl,
-NH-methallyl, -NHCHz-(furan-2-yl), -NHCHZ-cyclopropyl, -NH{t-butyl),
-NH(p-methylphenyl), -NHOCH3, -NHCHZ(p-fluoroghenyl), -NHCHZCHZOCH3,
-NH-cyclohexyl, -NHCHZCHZN(CH3)z, -NHCH,C(CH3)3, -NHCHZ-(pyrid-2-yl),
-NHCHZ-(pyrid-3-yI), -NHCHZ-(pyrid-4-yl), N-thiazolindinyl,
-N(CH~CHZCH3)~, -NHOH, -NH(p-NOZ-~), -NHCH,(p-NOZ-~),
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-NHCHZ(m-NOZ-~), -N(CH3)OCH3, -N(CH3)CHZ-~, -NHCHZ-(3,5-di-
fluorophenyl), -NHCHZCHZF, -NHCHZ(p-CH30-ø), -NHCH~(m-CH30-~),
-NHCHZ(p-CF3-~), -N(CH3)CH,CHZOCH3, -NHCH,CH2~, -NHCH(CH3)~,
-NHCH2-(p-F-~), -N(CH3)CH,CHZN(CH3)z, -NHCHZ-(tetrahydrofuran-2-yl),
and the like.
Still another preferred Y group is an alkyl group such as
-CH~CH,CH(CH3),, and the Iike.
Preferred heterocyclic structures defined by W and X include, by way of
example, 4-methylthiazolin-4-yl.
Preferred compounds of formula V and VI include the following:
(4R)-4-[N (1S)-(1-methoxycarbonyl-1-phenyl)methyl]carbamoyl-2-(3,5-
difluorophenylmethyl}-4-methyl-2-thiazoline
4-(N (S)-(1-methoxycarbonyl-1-phenyl)methyl]carbamoyl-2-(3,5-
difluorophenylmethyl}-2-thiazoline
4-[N (S)-(1-methoxycarbonyl-1-phenyl)methyl]carbamoyl-2-{3,5-
difluorophenylmethyl)-4-methyl-2-imidazoline
Yet another preferred compound of this invention includes lactams and
related compounds of formula VII and VIII
Rs
W_ H W
~ ~ T' N
R ~ C ~ H ~C(H)S VI I
~~ t
O
C
U
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-_
and
_ Rs
_ H W
R'~T'~ ~ N ~ ~ VII(
H ~ C~H)S
X X c ~ \C
U
wherein R', R'', R6) T' , X' , X", W and X are as defined above;
W' , together with -C(H)SC( =U)-, forms a cycloalkyl, cycloalkenyl,
heterocyclic, substituted cycloalkyl, or substituted cycloalkenyl group
wherein
each of said cycloalkyl, cycloalkenyl, heterocyclic, substituted cycloalkyl or
substituted cycloalkenyl group is optionally fused to form a bi- or mufti-
fused
ring system (preferably no more than 5 fused rings) with one or more ring
structures selected from the group consisting of cycloalkyl, cycloalkenyl,
heterocyclic, aryl and heteroaryl group which, in turn, each of such ring
structures is optionally substituted with 1 to 4 substituents selected from
the
group consisting of hydroxyl, halo, alkoxy, substituted alkoxy, thioalkoxy)
substituted thioalkoxy, aryl, heteroaryl, heterocyclic, nitro, cyano,
carboxyl,
carboxyl esters, alkyl, substituted alkyl, alkenyl, substituted alkenyl,
alkynyl,
substituted alkynyl, amino, N-alkylamino, N,N-dialkylamino, N-substituted
alkylamino, N-alkyl N-substituted alkylamino) N, N-disubstituted alkylamino,
-NHC(O)R'°, -NHSOZR'°, -C(O)NHz, -C(O)NHR'°, -
C(O)NR'°R'°, -S(O)Rlo,
-S(O),Rl°, -S(O)ZNHRI° and -S(O)~NR'°R'° where
each R'° is independently
selected from the group consisting of alkyl, substituted alkyl, or aryl;
U is selected from the group consisting of oxo (=O), thiooxo (=S),
hydroxyl (-H, -OH), thiol (H,-SH) and hydro (H,H);
s is an integer equal to 0 or 1; and
t is an integer equal to 0 or 1.
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-_ Zg __
In formula VII and VIII above, X" is preferably hydrogen, X' is
preferably hydrogen or fluoro or X' and X" form an oxo group, and T' is
preferably a covalent bond linking R' to -CX'X"-.
In formula VII and VIII above, preferred R' unsubstituted aryl groups
include, for example, phenyl, 1-naphthyl, 2-naphthyl, and the like.
Preferred Rl substituted aryl groups in formula VII and VIII include, for
example, monosubstituted phenyls (preferably 3 or 5 substituents);
disubstituted
phenyls (preferably 3,5 substituents); and trisubstituted phenyls (preferably
3,4,5 substituents). Preferably, the substituted phenyl groups do not include
more than 3 substituents.
Examples of substituted R' phenyls preferred in formula VII and VIII
include 2-chlorophenyl, 2-fluorophenyl, 2-bromophenyl, 2-hydroxyphenyl,
2-nitrophenyl, 2-methylphenyl, 2-methoxyphenyl, 2-phenoxyphenyl,
2-trifluoromethylphenyl, 4-fluorophenyl, 4-chlorophenyl,
4-bromophenyl, 4-nitrophenyl, 4-methylphenyl, 4-hydroxyphenyl,
4-methoxyphenyl, 4-ethoxyphenyl, 4-butoxyphenyl, 4-iso-propylphenyl,
4-phenoxyphenyl, 4-trifluoromethylphenyl, 4-hydroxymethylphenyl,
3-methoxyphenyl, 3-hydroxyphenyl, 3-nitrophenyl, 3-fluorophenyl,
3-chlorophenyl, 3-bromophenyl, 3-phenoxyphenyl, 3-thiomethoxyphenyl,
3-methylphenyl, 3-trifluoromethylphenyl, 2,3-dichlorophenyl,
2,3-difluorophenyl, 2,4-dichlorophenyl, 2,5-dimethoxyphenyl,
3,4-dichlorophenyl, 3,4-difluorophenyI, 3,4-methylenedioxyphenyl,
3,4-dimethoxyphenyl, 3,5-difluorophenyl, 3,5-dichlorophenyl, 3,5-di-
(trifluoromethyl)phenyl, 3,5-dimethoxyphenyl, 2,4-dichlorophenyl,
2,4-difluorophenyl, 2,6-difluorophenyl, 3,4,5-trifluorophenyl,
3,4,5-trimethoxyphenyl, 3,4,5-tri-(trifluoromethyl)phenyl, 2,4,6-
trifluorophenyl,
2,4,6-trimethylphenyl, 2,4,6-tri-(trifluoromethyl)phenyl, 2,3,5-
trifluorophenyl,
2,4,5-trifluorophenyl, 2,5-difluorophenyl, 2-fluoro-3-trifluoromethylphenyl,
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4-fluoro-2-trifluoromethylphenyl, 2-fluoro-4-trifluoromethylphenyl,
4-benzyloxyphenyl, 2-chloro-6-fluorophenyl, 2-fluoro-6-chlorophenyl,
2,3,4,5,6=pentafluorophenyl, 2,5-dimethylphenyl, 4-phenylphenyl and 2-fluoro-
3-trifluoromethylphenyl .
Preferred Rl alkaryl groups in formula VII and VIII include, by way of
example, benzyl, 2-phenylethyl, 3-phenyl-n-propyl, and the like.
Preferred Rl alkyl, substituted alkyl, alkenyl, cycloalkyl and
cycloalkenyl groups in formula VII and VIII include, by way of example,
iso-propyl, n-propyl, n-butyl, iso-butyl, see-butyl, t-butyl, -CH2CH=CH2,
-CH~CH=CH(CHZ)4CH~, cyclopropyl, cyclobutyl, cyclohexyl, cyclopentyl,
cyclohex-1-enyl, -CHI-cyclopropyl, -CHZ-cyclobutyl, -CHZ-cyclohexyl,
-CHZ-cyclopentyl, -CH,CHZ-cyclopropyl, -CHZCHZ-cyclobutyl, -CHZCH,-
cyclohexyl, -CHzCH2-cyclopentyl, and the like.
Preferred R' heteroaryls and substituted heteroaryls in formula VII and
VIII include, by way of example, pyrid-2-yl, pyrid-3-yI, pyrid-4-yl,
fluoropyridyls (including 5-fluoropyrid-3-yl), chloropyridyls (including
5-chloropyrid-3-yl), thiophen-2-yl, thiophen-3-yl, benzothiazol-4-yl,
2-phenylbenzoxazol-5-yl, furan-2-yl, benzofuran-2-yl, thionaphthen-2-yl,
2-chlorothiophen-5-yl, 3-methylisoxazol-5-yl, 2-(thiophenyl)thiophen-5-yl,
6-methoxythionaphthen-2-yl, 3-phenyl-1,2,4-thiooxadiazol-S-yI, 2-phenyloxazol-
4-yl, and the like.
Preferred R4 substituents in formula VII and VIII include, for example,
hydrogen, methyl, phenyl, benzyl and the like.
Preferred R6 substituents in formula VII and VIII include, for example,
hydrogen, methyl, ethyl, n-propyl, iso-propyl, n-butyl, iso-butyl, sec-butyl,
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tert-butyl, -CH2CH(CH,CH3)2, 2-methyl-n-butyl, 6-fluoro-n-hexyl-, phenyl,
benzyl, cyclohexyl) cyclopentyl) cycloheptyl, allyl, iso-but-2-enyl,
3-methylpentyI, -CHz-cyclopropyl, -CHZ-cyclohexyl, -CH,CH,-cyclopropyl)
-CHZCHZ-cyclohexyl, -CHZ-indol-3-yl) p-(phenyl)phenyl, o-fluorophenyl,
m-fluorophenyl, p-fluorophenyl, m-methoxyphenyl, p-methoxyphenyl,
phenethyl, benzyI, m-hydroxybenzyl, p-hydroxybenzyl, p-nitrobenzyl,
m-trifluoromethylphenyl, p-(CH3)~NCH~CHZCH,O-benzyl,
p-(CH3)3COC(O)CH20-benzyl, p-(HOOCCH20)-benzyl, 2-aminopyrid-6-yl,
p-(N-morphoiino-CH,CH,O)-benzyl, -CHZCH,C(O)NH~) -CH~-imidazol-4-yl,
-CHZ-(3-tetrahydrofuranyl), -CHZ-thiophen-2-yl, -CH,(1-methyl)cyclopropyl,
-CHI-thiophen-3-yl, thiophen-3-yl, thiophen-2-yl) -CHy-C(O)O-t-butyl,
-CH,-C(CH3)3, -CHzCH(CHZCH3)2, -2-methylcyclopentyl, -cyclohex-2-enyl,
-CH[CH(CH3)z]COOCH3, -CH,CH,N(CH3),, -CH,C(CH3)=CHZ,
-CHzCH =CHCH3 (cis and trans), -CHZOH, -CH(OH)CH3, -CH(O-t-butyl)CH3,
i5 -CH,OCI-13, -(CHZ).~NH-Boc, -(CH,)4NH2, -CHZ-pyridyl (e.g., 2-pyridyl,
3-pyridyl and 4-pyridyl), pyridyl (2-pyridyl, 3-pyridyl and 4-pyridyl),
-CH,-naphthyl (e.g., 1-naphthyl and 2-naphthyl), -CHZ-{N-morpholino),
p-(N-morpholino-CH,CH,O)-benzyl, benzo[b]thiophen-2-yl,
5-chlorobenzo[b]thiophen-2-yl, 4,5,6,7-tetrahydrobenzo[b]thiophen-2-yl,
benzo[b]thiophen-3-yl, 5-chlorobenzo[b]thiophen-3-yl, benzo[b]thiophen-5-yl,
6-methoxynaphth-2-yl, -CHZCH,SCH3, thien-2-yl, thien-3-yl, and the like.
Preferred cyclic groups defined by W' and -C(H)SC(--U)- include
cycloalkyl, Iactone, lactam, benzazepinone, dibenzazepinone and
benzodiazepine groups. In one preferred embodiment, the cyclic group defined
by W' and -C(H)SC(=U)-, forms a cycloalkyl group of the formula:
CH T«
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-- 31 --
wherein T" is selected from the group consisting of alkylene and substituted
alkylene.
A preferred cycloalkyl group is represented by the formula:
~~a~w
t
wherein each V is independently selected from the group consisting of hydroxy,
acyl, acyloxy, alkyl, substituted alkyl, alkoxy, substituted alkoxy, alkenyl)
substituted alkenyl, alkynyl, substituted alkynyl, amino, aminoacyl, alkaryl,
aryl, aryloxy, carboxyl, carboxylalkyl, cyano, halo, nitro, heteroaryl,
thioalkoxy, substituted thioalkoxy, trihalomethyl and the like; Ra is selected
from the group consisting of alkyl, substituted alkyl, alkoxy, substituted
alkoxy,
amino, carboxyl, carboxyl alkyl, cyano, halo, and the like; t is an integer
from
0 to 4; and w is an integer from 0 to 3.
Preferably t is an integer from 0 to 2 and, more preferably, is an integer
equal to 0 or 1.
In another preferred embodiment, the cyclic group defined by W')
together with -C(H)SC(=U)- is a ring of the formula:
T"
C(H)S
\CH
OH
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-- 32 --
or
-_
\CH
SH
wherein s is zero or one, T" is selected from the group consisting of
alkylene,
substituted alkylene, alkenylene, substituted alkenylene, -(R'3Z)qRl3- and -
ZR'3_
where Z is a substituent selected from the group consisting of -O-, -S- and
> NRIZ, each R'' is independently selected from the group consisting of alkyl,
alkenyl, alkynyl, cycioalkyl, cycloalkenyl, substituted alkyl, substituted
alkenyl,
substituted alkynyl, aryl, heteroaryl and heterocyclic, each R'3 is
independently
alkylene, substituted alkylene, alkenylene and substituted alkenylene with the
proviso that when Z is -O- or -S-, any unsaturation in the alkenylene and
substituted alkenylene does not involve participation of the -O- or -S-, and q
is
an integer of from 1 to 3.
Particularly preferred alcohol or thiol substituted groups include
N)
~Ra~w
i
lRa)w.
OH
OH
'~Ra)w
3$ HO
. _ . ~_.. ,
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-- 33 --
- Mr
w (v)~ _
s
(R )w (Ra)w
OH OH
wherein each V is independently selected from the group consisting of hydroxy,
acyl, acyloxy, alkyl, substituted alkyl, alkoxy, substituted alkoxy, alkenyl,
substituted alkenyl, alkynyl, substituted alkynyl, amino, aminoacyl, alkaryl,
aryl, aryloxy, carboxyl, carboxylalkyl, cyano, halo, vitro, heteroaryl,
thioalkoxy, substituted thioalkoxy, trihalomethyl and the like; Ra is selected
from the group consisting of alkyl, substituted alkyl, alkoxy, substituted
alkoxy,
1 s amino, carboxyl, carboxyl alkyl, cyano, halo, and the like; t is an
integer from
0 to 4; and w is an integer from 0 to 3.
Preferably t is an integer from 0 to 2 and, more preferably, is an integer
equal to 0 or 1.
Yet another preferred embodiment of the cyclic group defined by W',
together with -C(H)SC(=U}-, is a ring of the formula:
2s
C(H)S
\C
O
or
3s
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-_ 3~ __
T"
-
~C _
S
wherein s is zero or one, T" is selected from the group consisting of
alkylene,
substituted alkylene, alkenylene, substituted alkenylene, -(R'3Z)qR'3- and -
ZR'3-
where Z is a substituent selected from the group consisting of -O-, -S- and
> NR'', each Rl' is independently selected from the group consisting of alkyl,
alkenyl, alkynyl, cycloalkyl, cycloalkenyl, substituted alkyl, substituted
alkenyl,
substituted alkynyl, aryl, heteroaryl and heterocyclic, each R'3 is
independently
alkylene, substituted alkylene, alkenylene and substituted alkenylene with the
proviso that when Z is -O- or -S-, any unsaturation in the alkenylene and
substituted alkenylene does not involve participation of the -O- or -S-, and q
is
an integer of from 1 to 3.
Particularly preferred cyclic ketone and thioketone groups include:
~Ra)w
{U)t
(~)t
O
(Ra)w / ~ (~t
'(Ra)w
O
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-- 35 --
wherein each V is independently selected from the group consisting of hydroxy,
acyl, acyloxy, alkyl, substituted alkyl, alkoxy, substituted alkoxy, alkenyl,
substituted alkenyl, alkynyl, substituted alkynyl) amino, aminoacyl, alkaryl)
aryl, aryloxy, carboxyl, carboxylalkyl, cyano, halo, vitro, heteroaryl,
thioalkoxy, substituted thioalkoxy, trihalomethyl and the like; Ra is selected
from the group consisting of alkyl, substituted alkyl) alkoxy, substituted
alkoxy,
amino, carboxyl, carboxyl alkyl, cyano, halo, and the like; t is an integer
from
0 to 4; and w is an integer from 0 to 3.
Preferably t is an integer from 0 to 2 and, more preferably, is an integer
equal to 0 or 1.
In another preferred embodiment, the cyclic group defined by W' ,
together with -C(H)SC(=U)-) forms a ring of the formula:
T"
C(H)S
~C /O
fl
0
T"
C(H)S
/S
C
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-- 36 --
T'
--
O
s ~c -i _
s
to or
T"
WH)S
is ~C /S
S
wherein s is zero or one, T" is selected from the group consisting of
alkylene,
substituted alkylene, alkenylene, substituted alkenylene, -(R"Z)QR'3- and -
ZR'3-
where Z is a substituent selected from the group consisting of -O-, -S- and
> NR'z, each R'2 is independently selected from the group consisting of alkyl,
alkenyl, alkynyl, cycloalkyl, cycloalkenyl, substituted alkyl, substituted
alkenyl,
substituted alkynyl, aryl, heteroaryl and heterocyclic, each R'3 is
independently
alkylene, substituted alkyiene, alkenylene and substituted alkenylene with the
proviso that when Z is -O- or -S-, any unsaturation in the alkenylene and
substituted alkenylene does not involve participation of the -O- or -S-, and q
is
an integer of from 1 to 3.
Particularly preferred lactone and thiolactone groups include:
~V)t
~Ra)w
(R )w
i
0
and
O O
O
T, .. ~ . ....
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__ 3~ __
wherein each V is independently selected from the group consisting of hydroxy,
acyl) acyloxy) alkyl, substituted alkyl, alkoxy, substituted alkoxy, alkenyl,
substituted aikenyi, alkynyl, substituted alkynyl, amino, aminoacyl, alkaryl)
aryl, aryloxy, carboxyl, carboxylalkyl, cyano, halo, vitro, heteroaryl,
thioalkoxy, substituted thioalkoxy, trihalomethyl and the like; Ra is selected
from the group consisting of alkyl, substituted alkyl, alkoxy, substituted
alkoxy,
amino, carboxyl, carboxyl alkyl, cyano, halo, and the like; t is an integer
from
0 to 4; and w is an integer from 0 to 3.
Preferably t is an integer from 0 to 2 and ) more preferably, is an integer
equal to 0 or 1.
In another preferred embodiment, the cyclic group defined by W' and
-C(H)SC(=U)-, forms a lactam ring of the formula:
T"
~ / NR~z
C
O
or a thiolactam ring of the formula:
T'
~C / NR~z
I I
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__ 3g __
wherein s is zero or one, T" is selected from the group consisting- of
alkylene,
substituted alkylene, alkenylene, substituted alkenylene, -(R'3Z)qR'3- and -
ZR'3-
where Z is a substituent selected from the group consisting of -O-, -S- and
> NR'Z, each R'Z is independently selected from the group consisting of alkyl,
alkenyl, alkynyl, cycloalkyl, cycloalkenyl, substituted alkyl, substituted
alkenyl,
substituted alkynyl, aryl, heteroaryl and heterocyclic, each R'3 is
independently
alkylene, substituted alkylene, alkenylene and substituted alkenylene with the
proviso that when Z is -O- or -S-, any unsaturation in the alkenylene and
substituted alkenylene does not involve participation of the -O- or -S-, and q
is
an integer of from 1 to 3.
Particularly preferred Iactam and thiolactam groups include:
(V)t
(Ra)w
N ~Rb N~ Rb
O
O
(Ra)...
~V)t
(V)
(Ra)w
Ra \
i
N Rb N~Rb
O O
t
~.. ' _..
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__ 3g __
rv~, (~)t, Mt,
i
' ~N~Rb
(Ra)w (Ra)w
~--
J
p~N~Rb Nv
O Rb
(R )w (Ra)w (~t
N
O \Rb
O ~Rb
(V)t
(Ra)w
~Ra)w
N ~N.
O ~Rb O ~Rb
Rb
(V)t
\R8)W Q ~ O N
. 30 N ~ ~
O~ \Rb N \
O Rb
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-- 40 --
s
R~
Rb
N
N ~ ' W)c
/ ~V)t N /
~ O~ ~5
Rb
AFB
N
N ~ \ N w
is ~ / ~~)t ~ , c
O
wherein A-B is selected from the group consisting of alkylene, alkenylene,
substituted alkylene, substituted alkenylene and -N =CH-; Q' is oxygen or
sulfur; each V is independently selected from the group consisting of hydroxy,
acyl, acyloxy, alkyl, substituted alkyl, alkoxy) substituted alkoxy, alkenyl,
substituted alkenyl, alkynyl, substituted alkynyl, amino, aminoacyl, alkaryl,
aryl, aryloxy, carboxyl, carboxylalkyl, cyano, halo, vitro, heteroaryl,
thioalkoxy, substituted thioalkoxy, trihalomethyl and the like; Ra is selected
from the group consisting of alkyl, substituted alkyl, alkoxy, substituted
alkoxy,
,.- , ..._
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-- 41 --
amino, carboxyl) carboxyl alkyl, cyano, halo, and the like; Rb is selected
from
the group consisting of hydrogen, alkyl, substituted alkyl, alkenyl,
substituted
alkenyl, alkynyl, substituted alkynyl, acyI) aryl, heteroaryl, heterocyclic,
and
the like; R' is selected from the group consisting of alkyl, substituted
alkyl)
alkenyl, substituted alkenyl, aryl, heteroaryl, heterocyclic, cycloalkyl, and
substituted cycloalkyl; t is an integer from 0 to 4; t' is an integer from 0
to 3;
and w is an integer from 0 to 3.
Preferably t is an integer from 0 to 2 and, more preferably, is an integer
equal to 0 or 1.
In another preferred embodiment, the cyclic group defined by W',
together with -C(H)SC(=U)-, forms a ring of the formula:
T"
C~H)s
NR~2
CH2
wherein s is zero or one, T" is selected from the group consisting of
alkylene,
substituted alkylene, alkenylene, substituted alkenylene, -(R13Z)qR'3- and -
ZR'3-
where Z is a substituent selected from the group consisting of -O-, -S- and
> NR'Z, each R'' is independently selected from the group consisting of alkyl,
alkenyl, alkynyl, cycloalkyl, cycloalkenyl, substituted alkyl, substituted
alkenyl,
substituted alkynyl, aryl, heteroaryl and heterocyclic, each R'3 is
independently
alkylene, substituted alkylene, alkenylene and substituted alkenylene with the
proviso that when Z is -0- or -S-, any unsaturation in the alkenylene and
substituted alkenylene does not involve participation of the -O- or -S-, and q
is
an integer of from 1 to 3.
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A still further preferred embodiment is directed to a ring group defined
by W, together with -C(H)SC(=U)-, of the formula:
C~H)S
\CH
NRt2R'2
wherein s is zero or one, T" is selected from the group consisting of
alkylene,
substituted alkylene, alkenylene, substituted alkenylene, -(R'3Z)qR'3- and -
ZR"-
where Z is a substituent selected from the group consisting of -O-, -S- and
> NR'Z, each RIZ is independently selected from the group consisting of alkyl,
aikenyl) alkynyl, cycloalkyl, cycloalkenyl, substituted alkyl, substituted
alkenyl,
substituted alkynyl, aryl, heteroaryl and heterocyclic, each R13 is
independently
alkylene, substituted alkylene, alkenylene and substituted alkenylene with the
proviso that when Z is -O- or -S-, any unsaturation in the alkenyiene and
substituted alkenylene does not involve participation of the -O- or -S-, and q
is
an integer of from 1 to 3.
Yet another preferred compound of this invention includes lactams and
related compounds of formula IX and X:
O
W X
R, N ~ /C(H)S IX
~~ N
L 'Z J m N a H ~C
R2 R
U
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-- 43 --
and
- p /W
S R N N X N/C(Fi) X
\C
R
Rz
U
wherein R', R', R4, Z, U, W, W', X, m and s are as defined above.
Preferred R', R'-, R°, Z, W and W' are also as defined above.
In one of its method aspects, this invention is directed to a method for
inhibiting ~3-amyloid peptide release and/or its synthesis in a cell which
method
comprises administering to such a cell an amount of a compound or a mixture of
compounds of formula I, VII and VIII above effective in inhibiting the
cellular
release and/or synthesis of ~3-amyloid peptide.
Because the in vivo generation of /3-amyloid peptide is associated with the
pathogenesis of ADg~9, the compounds of formula I, VII and VIII can also be
employed in conjunction with a pharmaceutical composition to prophylactically
and/or therapeutically prevent and/or treat AD. Accordingly, in another of its
method aspects, this invention is directed to a prophylactic method for
preventing
the onset of AD in a patient at risk for developing AD which method comprises
administering to said patient a pharmaceutical composition comprising a
pharmaceutically inert carrier and an effective amount of a compound or a
mixture
of compounds of formula I, VII and VIII above.
In yet another of its method aspects, this invention is directed to a
therapeutic method for treating a patient with AD in order to inhibit further
deterioration in the condition of that patient which method comprises
administering
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to said patient a pharmaceutical composition comprising a phan naceutically
inert
carrier and an effective amount of a compound or a mixture of compounds of
formula I,-VII and VIII above.
This invention also provides for novel pharmaceutical compositions
comprising a pharmaceutically inert carrier and a compound of the formula I,
VII
or VIII above.
Still further, this invention provides for novel compounds of formula I, VII
or VIII.
Further preferred compounds of formula I are represented in the following
Tables I-IV which illustrate preferred subsets of these structures:
_...~.,..~.~.~._ ... . _ ?
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-- 45 --
TABLE T
C(O)Y
H ~
R~~/N ~N~
Ra
R2
R'' R' Ra Y W X
3,5-di-F-~-CH~C(O)--CH3 H -OCH~CH3 S -CHz-
~-CH,-OC(O)- -CH3 -CH,c -OCH3 -N-t-Boc -
C H,-
3,5-di-F-~-CH,C(O)--CH3 -CH2~ -OCH3 -N-t-Boc -CHZ-
3,5-di-F-~-CHZC(O)--CH3 -CH~cp -OCH3 NH -CHZ-
~-CH,OC(O)- -CH3 -~ -OCH3 -N-t-Boc -CH,-
3,5-di-F-~-CHZC(O)--CH3 -~ -OCH3 NH -CHZ-
3,5-di-F-~-CHZC(O)--CH3 H -OCH3 O -CHz-
3,5-di-Cl-~-CHZC(O)--CH3 H -OCH3 O -CHZ-
3,5-di-F-~-CH~C(O)--~ H -OCH3 S -CHZ-
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-- 46 --
TABLE II -
~X C(O)Y
R~' ~ VV'
Ra
R2
Ri . Rz R.~ Y W X
3,5-difluoro-~-CHz-C{O)--CH3 H -OCHzCH3 O -CHz-
3,5-difluoro-~-CHz-C(O)--CH3 -CHz~ -OCH3 NH -CHz-
3,5-difluoro-~-CHz-C(O)--CH3 -~ -OCH3 NH -CHz-
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__ 4~ __
TABLE III
Rs
H O
,~N
R~ wN wV
J H
R2 m
R'' R'- m R6 V
3,5-di-F-~-CHZC(O)- -CH3 1 -CH3 3-ethyl-1,2,4-oxadiazol-
5-yl
3,5-di-F-~-CHZC(O)- -CH3 1 -CHz~ 3-methyl-1,2,4-
oxadiazol-S-yl
3, 5-di-F-~-CH~C(O)- -- 0 -CH3 1-(methylhexano-
2-yl)-
tetrazol-S-yl
3, S-di-F-~-CH2C(O)- -CH3 1 -~ thiazolin-2-yl
3,5-di-F-~-CHzC(O)- -CH3 1 -~ 3-methyl-1,2,4-
oxadiazol-S-yl
3,5-di-F-~-CHZC(O)- -CH3 I -~ 3-phenyl-1,2,4-
oxadiazol-S-yl
3,5-di-F-~-CHzC(O)- -CH3 1 -~ 3-(p-methoxy-benzyl)-
~ ~ ~ 1,2,4-oxadiazol-5-yl
~
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__ 4g __
TABLE I V
0 R2
Y
R~,-T .N
H
n
RI T n R
3,5-di-F-~-CHI- 2-R'-thiazolin-4-yl 1 -~ -OCH3
3,5-di-F-~-CHZ- 2-R'-4-methyl- 1 -~ -OCH3
thiazolin-4-yl
3,5-di-F-~-CHZ- 2-R'-4-methyl- 1 -~ -OCH3
imidazolin-4-yl
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DETAILED DESCRIPTION OF THE INVENTION
As above, this invention relates to compounds which inhibit a-amyloid
peptide release and/or its synthesis, and, accordingly, have utility in
treating
Alzheimer's disease. However, prior to describing this invention in further
detail,
the following terms will first be defined.
Definitions
The term "~3-amyloid peptide" refers to a 39-43 amino acid peptide having
a molecular weight of about 4.2 kD, which peptide is substantially homologous
to
the form of the protein described by Glenner, et al.' including mutations and
post-
translational modifications of the normal ~3-amyloid peptide. In whatever
form,
the ~3-amyloid peptide is an approximate 39-43 amino acid fragment of a large
membrane-spanning glycoprotein, referred to as the ~3-amyloid precursor
protein
(APP). Its 43-amino acid sequence is:
_1
Asp Ala Glu Phe Arg His Asp Ser Gly Tyr
_11
Glu Val His His Gln Lys Leu Val Phe Phe
_21
Ala Glu Asp Val Gly Ser Asn Lys Gly Ala
_31
Ile Ile Gly Leu Met Val Gly Gly Val Val
41
Ile Ala Thr (SEQ ID NO: 1)
or a sequence which is substantially homologous thereto.
"Alkyl" refers to monovalent alkyl groups preferably having from 1 to 10
carbon atoms and more preferably 1 to 6 carbon atoms. This term is exemplified
by groups such as methyl, ethyl, n-propyl, iso-propyl, n-butyl, iso-butyl, n-
hexyl,
and the like.
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"Substituted alkyl" refers to an alkyl group, preferably of from 1 to 10
carbon atoms, having from 1 to 3 substituents selected from the group
consisting
of alkoxy,-substituted alkoxy, acyl, acylamino, amino, aminoacyl, aminocarboxy
esters, cyano, cycloalkyl, halogen, hydroxyl, carboxyl, carbaxylalkyl,
oxyacyl,
oxyacylamino, thiol, thioalkoxy, substituted thioalkoxy, aryl, heteroaryl,
heterocyclic, aryloxy, thioaryloxy, heteroaryloxy, thioheteroaryloxy, nitro,
and
mono- and di-alkylamino, mono- and di-(substituted alkyl)amino, mono- and di-
arylamino, mono- and di-heteroarylamino, mono- and di-heterocyclic amino, and
unsymmetric di-substituted amines having different substituents selected from
alkyl, substituted alkyl, aryl, heteroaryl and heterocyclic.
"Alkylene" refers to divalent alkylene groups preferably having from 1 to
10 carbon atoms and more preferably 1 to 6 carbon atoms. This term is
exemplified by groups such as methylene (-CHZ-), ethylene (-CHZCHZ-), the
propylene isomers (e.g., -CHZCHZCHZ- and -CH(CH3)CHZ-), and the like.
"Alkaryl" refers to -alkylene-aryl groups preferably having from 1 to 10
carbon atoms in the alkylene moiety and from 6 to 10 carbon atoms in the aryl
moiety. Such alkaryl groups are exemplified by benzyl, phenethyl and the like.
"Alkoxy" refers to the group "alkyl-O-". Preferred alkoxy groups include,
by way of example, methoxy, ethoxy, n-propoxy, iso-propoxy,
n-butoxy, tert-butoxy, sec-butoxy, n-pentoxy, n-hexoxy, 1,2-dimethylbutoxy,
and
the like.
"Substituted alkoxy" refers to the group "substituted alkyl-O-" where
substituted alkyl is as defined above.
"Alkenyl" refers to alkenyl groups preferably having from 2 to 10 carbon
atoms and more preferably 2 to 6 carbon atoms and having at least 1 and
.r ~ ~
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preferably from 1-2 sites of alkenyl unsaturation. Preferred alkenyl groups
include
ethenyl (-CH=CHI), n-propenyl (-CH,CH=CH,), iso-propenyl
(-C(CH;)=CHI), but-2-enyl (-CH~CH=Cl-1CH;), and the like.
"Substituted alkenyl" refers to an alkenyl group as defined above having
from 1 to 3 substituents selected from the group consisting of alkoxy,
substituted
alkoxy, acyl, acylamino, amino, aminoacyl, aminocarboxy esters, cyano,
halogen,
hydroxyl, carboxyl, carboxylalkyl, cycloalkyl, oxyacyl, oxyacylamino, thiol,
thioalkoxy, substituted thioalkoxy, aryl, heteroaryl, heterocyclic, nitro, and
mono-
and di-alkylamino, mono- and di-(substituted alkyl)amino, mono- and di-
arylamino,
mono- and di-heteroarylamino, mono- and di-heterocyclic amino, and unsymmetric
di-substituted amines having different substituents selected from alkyl,
substituted
alkyl, aryl, heteroaryl and heterocyclic.
1 S "Alkenylene" refers to divalent alkenylene groups preferably having from 2
to 8 carbon atoms and more preferably 2 to 6 carbon atoms. This term is
exemplified by groups such as ethenylene (-CH=CH-), the propenylene isomers
(e.g., -CH,CH=CH- and -C(CH3)=CH-) and the like.
"Substituted alkenylene" refers to an alkenylene group, preferably of from 2
to 8 carbon atoms, having from 1 to 3 substituents selected from the group
consisting of alkoxy, substituted alkoxy, acyl, acylamino, acyloxy, amino,
aminoacyl, aminoacyloxy, cyano, halogen, hydroxyl, carboxyl, carboxylalkyl,
thiol,
thioalkoxy, substituted thioalkoxy, aryl, heteroaryl, heterocyclic, nitro, and
mono-
and di-alkylamino, mono- and di-(substituted alkyl)amino, mono- and di-
arylamino,
mono- and di-heteroarylamino, mono- and di-heterocyclic amino, and unsymmetric
di-substituted amines having different substituents selected from alkyl,
substituted
alkyl, aryl, heteroaryl and heterocyclic. Additionally, such substituted
alkylene
groups include those where 2 substituents on the alkylene group are fused to
form
one or more cycloalkyl, aryl, heterocyclic or heteroaryl groups fused to the
alkylene group.
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"Alkynyl" refers to alkynyl groups preferably having from-2 to 10 carbon
atoms and more preferably 2 to 6 carbon atoms and having at least 1 and
preferably-from 1-2 sites of alkynyl unsaturation. Preferred alkynyl groups
include
ethynyl (-C---CH), propargyl (-CH,C---CH) and the like.
"Substituted alkynyl" refers to an alkynyl group as defined above having
from 1 to 3 substituents selected from the group consisting of alkoxy,
substituted
alkoxy, acyl, acylamino, amino, aminoacyl, aminocarboxy esters, cyano,
halogen,
hydroxyl, carboxyl, carboxylalkyl, cycloalkyl, oxyacyl, oxyacylamino, thiol,
thioalkoxy, substituted thioalkyoxy, aryl, heteroaryl, heterocyclic, vitro,
and mono-
and di-alkylamino, mono- and di-(substituted alkyl)amino, mono- and di-
arylamino,
mono- and di-heteroarylamino, mono- and di-heterocyclic amino, and unsymmetric
di-substituted amines having different substituents selected from alkyl,
substituted
alkyl, aryl, heteroaryl and heterocyclic.
"Acyl" refers to the groups alkyl-C{O)-, substituted alkyl-C(O)-, cycloalkyl-
C(O)-, aryl-C(O)-, heteroaryl-C(O)- and heterocyclic-C(O)- where alkyl,
substituted
alkyl, cycloalkyl, aryl, heteroaryl and heterocyclic are as defined herein.
"Acylamino" refers to the group -C{O)NRR where each R is independently
hydrogen, alkyl, substituted alkyl, cycloalkyl, aryl, heteroaryl, and
heterocyclic and
where each of alkyl, substituted alkyl, cycloalkyl, aryl, heteroaryl and
heterocyclic
are as defined herein.
"Aminoacyl" refers to the group -NRC(O)R where each R is independently
hydrogen, alkyl, substituted alkyl, cycloalkyl, aryl, heteroaryl, and
heterocyclic and
where each of alkyl, substituted alkyl, cycloalkyl, aryl, heteroaryl and
heterocyclic
are as defined herein.
_. .._....e.....~..~. ~.rc~... T
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"Oxyacyl" refers to the groups -OC(O)-alkyl, -OC(O)-aryl, -C(O)O-
heteroaryl-, and -C(O)O-heterocyclic where alkyl, aryl, heteroaryl and
heterocyclic
are as del=fined herein.
S "Oxyacylamino" refers to the groups -OC(O)NR-alkyl, -OC(O}NR-
substituted alkyl, -OC(O)NR-aryl, -OC(O)NR-heteroaryl-, and -OC(O)NR-
heterocyclic where R is hydrogen, alkyl, substituted alkyl, cycloalkyl, aryl,
heteroaryl, and heterocyclic and where each of alkyl, substituted alkyl,
cycloalkyl,
aryl, heteroaryl and heterocyclic are as defined herein.
"Aminocarboxy esters" refers to the groups -NRC(O)O-alkyl,
-NRC(O)O-substituted alkyl, -NRC(O)O-aryl, -NRC(O)O-heteroaryl, and
-NRC(O)O-heterocyclic where R is hydrogen, alkyl, substituted alkyl,
cycloalkyl,
aryl, heteroaryl, and heterocyclic and where each of alkyl, substituted alkyl,
cycloalkyl, aryl, heteroaryl and heterocyclic are as defined herein.
"Aryl" refers to an unsaturated aromatic carbocyclic group of from 6 to 14
carbon atoms having a single ring (e.g., phenyl) or multiple condensed rings
(e.g.,
naphthyl or anthryl). Preferred aryls include phenyl, naphthyl and the like.
Unless otherwise constrained by the definition for the aryl substituent, such
aryl groups can optionally be substituted with from 1 to 3 substituents
selected
from the group consisting of hydroxy, acyl, alkyl, alkoxy, alkenyl, alkynyl,
substituted alkyl, substituted alkoxy, substituted alkenyl, substituted
alkynyl, amino,
aminoacyl, aminocarboxy esters, alkaryl, aryl, aryloxy, carboxyl,
carboxylalkyl,
acylamino, cyano, halo, nitro, heteroaryl, heterocyclic, oxyacyl,
oxyacylamino,
thioalkoxy, substituted thioalkoxy, trihalomethyl, mono- and di-alkylamino,
mono-
and di-(substituted alkyl)amino, mono- and di-arylamino, mono- and di-
heteroarylamino, mono- and di-heterocyclic amino, and unsymmetric di-
substituted
amines having different substituents selected from alkyl, substituted alkyl,
aryl,
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heteroaryl and heterocyclic, and the like. Preferred substituents include
alkyl,
alkoxy, halo, cyano, nitro, trihalomethyl, and thioalkoxy.
"Aryloxy" refers to the group aryl-O- wherein the aryl group is as defined
above including optionally substituted aryl groups as also defined above.
"Carboxyalkyl" refers to the groups -C(O}O-alkyl and -C{O)O-substituted
alkyl where alkyl and substituted alkyl are as defined above.
"Cycloalkyl" refers to cyclic alkyl groups of from 3 to 8 carbon atoms
having a single cyclic ring or multiple condensed rings which can be
optionally
substituted with from 1 to 3 alkyl groups. Such cycloalkyl groups include, by
way
of example, single ring structures such as cyclopropy l, cyclobutyl,
cyclopentyl,
cyclooctyl, I -methylcyclopropyl, 2-methylcyclopentyl, 2-methylcyclooctyl, and
the
like, or multiple ring structures such as adamantanyl, and the like.
"Substituted cycloalkyl" refers to cycloalkyl groups having from 1 to 5
(preferably 1 to 3) substituents selected from the group consisting of
hydroxy, acyl,
acyloxy, alkyl, substituted alkyl, alkoxy, substituted alkoxy, alkenyl,
substituted
alkenyl, alkynyl, substituted alkynyl, amino, aminoacyl, alkaryl, aryl,
aryloxy,
carboxyl, carboxylalkyi, cyano, halo, nitro, heteroaryl, thioalkoxy,
substituted
thioalkoxy, trihalomethyl and the like.
"Cycloalkenyl" refers to cyclic alkenyl groups of from 4 to 8 carbon atoms
having a single cyclic ring and at least one point of internal unsaturation
which can
be optionally substituted with from 1 to 3 alkyl groups. Examples of suitable
cycloalkenyl groups include, for instance, cyclobut-2-enyl, cyclopent-3-enyl,
cyclooct-3-enyl and the like.
"Substituted cycloalkenyl" refers to cycloalkenyl groups having from 1 to 5
substituents selected from the group consisting of hydroxy, acyl, acyloxy,
alkyl,
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-- 5 5 --
substituted alkyl, alkoxy, substituted alkoxy, alkenyl, substituted aikenyl,
alkynyl,
substituted alkynyl, amino, aminoacyl, alkaryl, aryl, aryloxy, carboxyl,
carboxylalkyl, cyano, halo, nitro, heteroaryl, thioalkoxy, substituted
thioalkoxy,
trihalomethyl and the like. -
"Halo" or "halogen" refers to fluoro, chloro, bromo and iodo and preferably
is either chloro or bromo.
"Heteroaryl" refers to a monovalent aromatic carbocyclic group of from 2 to
10 carbon atoms and 1 to 4 heteroatoms selected from oxygen, nitrogen and
sulfur
within the ring.
Unless otherwise constrained by the definition for the heteroaryl substituent,
such heteroaryl groups can be optionally substituted with 1 to 3 substituents
selected from the group consisting of hydroxy, acyl, alkyl, alkoxy, alkenyl,
alkynyl,
substituted alkyl, substituted alkoxy, substituted alkenyl, substituted
alkynyl, amino,
aminoacyl, aminocarboxy esters, alkaryl, aryl, aryloxy, carboxyl,
carboxylalkyI,
aminoacyl, cyano, halo, nitro, heteroaryl, heterocyclic, oxyacyl,
oxyacylamino,
thioalkoxy, substituted thioaIkoxy, trihalomethyl, mono- and di-alkylamino,
mono-
and di-(substituted alkyl}amino, mono- and di-arylamino, mono- and di-
heteroarylamino, mono- and di-heterocyclic amino) and unsymmetric di-
substituted
amines having different substituents selected from alkyl, substituted alkyl,
aryl,
heteroaryl and heterocyclic, and the like. Preferred substituents include
alkyl,
alkoxy, halo, cyano, nitro, trihalomethyl) and thioalkoxy.
Such heteroaryl groups can have a single ring (e.g., pyridyl or furyl) or
multiple condensed rings (e.g., indolizinyl or benzothienyl}. Preferred
heteroaryls
include pyridyl, pyrrolyl and furyl.
"Heteroeycle" or "heterocyclic" refers to a monovalent saturated or
unsaturated group having a single ring or multiple condensed rings, from 1 to
8
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carbon atoms and from I to 4 hetero atoms selected from nitrogeij, sulfur or
oxygen within the ring.
Unless otherwise constrained by the definition for the-heterocyclic
substituent, such heterocycIic groups can be optionally substituted with 1 to
3
substituents selected from the group consisting of hydroxy, acyl, alkyl,
alkoxy,
alkenyl, alkynyl, substituted alkyl, substituted alkoxy, substituted alkenyl,
substituted alkynyl, amino, aminoacyl, aminocarboxy esters, alkaryl, aryl,
aryloxy,
carboxyl, carboxylalkyl, aminoacyl, cyano, halo, nitro, heteroaryl,
heterocyclic,
oxyacyl, oxyacylamino, thioalkoxy, substituted thioalkoxy, trihalomethyl, mono-
and di-alkylamino, mono- and di-{substituted alkyl)amino, mono- and di-
arylamino,
mono- and di-heteroarylamino, mono- and di-heterocyclic amino, and unsymmetric
di-substituted amines having different substituents selected from alkyl,
substituted
alkyl, aryl, heteroaryl and heterocyclic, and the like. Such heterocyclic
groups can
have a single ring or multiple condensed rings. Preferred heterocyclics
include
morpholino, piperidinyl, and the like.
Examples of heterocycles and heteroaryls include, but are not limited to,
pyrrole, imidazole, pyrazole, pyridine, pyrazine, pyrimidine, pyridazine,
indolizine,
isoindole, indole, indazole, purine, quinolizine, isoquinoline, quinoline,
phthalazine,
naphthylpyridine, quinoxaline, quinazoline, cinnoline, pteridine, carbazole,
carboline, phenanthridine, acridine, phenanthroline, isothiazole, phenazine,
isoxazole, phenoxazine, phenothiazine, imidazolidine, imidazoline, piperidine,
piperazine, indoline, phthalimide, 1,2,3,4-tetrahydroisoquinoline, 4,5,6,7-
tetrahydrobenzo[b]thiophene, thiazole, thiazolidine, thiophene,
benzo[b]thiophene,
morpholino, piperidinyl, pyrrolidine, tetrahydrofuranyl, and the like.
"Oxyacyl" refers to the groups -OC(O)-alkyl, -OC(O)-aryl, -C(O)O
heteroaryl-, and -C(O)O-heterocyclic where alkyl, aryl, heteroaryl and
heterocyclic
are as defined herein.
. ... ~.." ~
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__
"Oxyacylamino" refers to the groups -OC(O)NH-alkyl, -OC(O)NH-
substituted alkyl, -OC(O)NH-aryl, -OC(O)NH-heteroaryl-, and -OC(O)NH-
heterocyclic where alkyl, aryl, heteroaryl and heterocyclic are as defined
herein.
"Thiol" refers to the group -SH.
"Thioalkoxy" refers to the group -S-alkyl.
"Substituted thioalkoxy" refers to the group -S-substituted alkyl.
"Thioaryloxy" refers to the group aryl-S- wherein the aryl group is as
defined above including optionally substituted aryl groups also defined above.
"Thioheteroaryloxy" refers to the group heteroaryl-S- wherein the heteroaryl
group is as defined above including optionally substituted aryl groups as also
defined above.
"Pharmaceutically acceptable salt" refers to pharmaceutically acceptable salts
of a compound of formula I, VII or VIII which salts are derived from a variety
of
organic and inorganic counter ions well known in the art and include, by way
of
example only, sodium, potassium, calcium, magnesium, ammonium,
tetraalkylammonium, and the Like; and when the molecule contains a basic
functionality, salts of organic or inorganic acids, such as hydrochloride,
hydrobromide, tartrate, mesylate, acetate, maleate, oxalate and the like.
Compound Preparation
The compounds of formula I are readily prepared via several divergent
synthetic routes with the particular route selected relative to the ease of
compound
preparation, commercial availability of starting materials, etc.
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__ 5g __
The heterocyclic rings) in the compounds of formula I above can be
prepared by use or adaptation of known chemical syntheses and methods, which
synthesis 'and methods are well known in the art. For example, the synthesis
of
various heterocycles is illustrated in detail in the examples set forth below.
Using
these procedures, other suitable heterocycles can be prepared by modifying the
starting materials employed in these procedures.
In certain cases, the compounds of formula I above are prepared by
coupling a suitable heterocycle having one or more pendant functional groups
to
the other components necessary to form the A-B-C structure of formula I.
Typical
functional groups used for such couplings include, by way of example,
carboxylic
acid and amino groups. The coupling reaction is generally conducted using
conventional coupling reagents such as carbodiimides with or without the use
of
well known additives such as N-hydroxysuccinimide, 1-hydroxybenzotriazole,
etc.
can be used to facilitate coupling. The reaction is typically conducted in an
inert
aprotic polar diluent such as dimethylformamide, dichloromethane, chloroform,
acetonitrile, tetrahydrofuran and the like.
The compounds of formula VII and VIII are readily prepared by
conventional amidation of a carboxylic acid as shown in reaction ( 1 ) below:
~.. ~
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O
W X
Rf iT ~ C ~ N H 1a
/ \ N ~R4 ~ H
X' X"
t
or
Rs
N-
R~.~T'~~ ~ N OH 1b
/ \ H ~I
x' x" o
-t- ,
H2N--C(H)S 2
\C
U
Reaction 1
VI I (for 1 a) or VI II (for 1 b)
wherein R', R4, R6, T', X', X", W, X, W', U, t and s are as defined above.
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The reaction is conventionally conducted by using at least -a
stoichiometric amount of carboxylic acid la or lb and amine 2. This reaction
is conventionally conducted for peptide synthesis and synthetic methods used
therein can also be employed to prepare compounds VII and VIII. For
example, well known coupling reagents such as carbodiimides with or without
the use of well known additives such as N-hydroxysuccinimide,
1-hydroxybenzotriazole, etc. can be used to facilitate coupling. The reaction
is
conventionally conducted in an inert aprotic polar diluent such as
dimethylformamide, dichioromethane, chloroform, acetonitrile, tetrahydrofuran
and the like. Alternatively, the acid halide of compound la or lb can be
employed in reaction (1 ) and, when so employed, it is typically employed in
the
presence of a suitable base to scavenge the acid generated during the
reaction.
Suitable bases include, by way of example, triethylamine,
diisopropylethylamine, N-methylmorpholine and the like.
Synthesis of Carboxylic Acid Starting materials
Carboxylic acids la and lb can be prepared by several divergent
synthetic routes with the particular route selected relative to the ease of
compound preparation, commercial availability of starting materials, whether t
is zero or one. One method for preparing these compounds (when t is one) is
the hydrolysis of esters of compounds of formula V and VI above. Similar
methods can be employed to prepare related compounds when t is zero.
Carboxylic acids 1 having m equal to 1 and n equal to 1 or 2 can also be
prepared by use of polymer supported forms of carbodiimide peptide coupling
reagents. A polymer supported form of EDC, for example, has been described
(Tetrahedron Letters, 34(48}, 7685 ( 1993})'°. Additionally, a new
carbodiimide
coupling reagent, PEPC, and its corresponding polymer supported forms have
been discovered and are very useful for the preparation of such compounds.
r. ~
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Polymers suitable for use in making a polymer supported coupling
reagent are either commercially available or may be prepared by methods well
" known to the artisan skilled in the polymer arts. A suitable polymer must
possess pendant sidechains bearing moieties reactive with the -terminal amine
of
the carbodiimide. Such reactive moieties include chloro, bromo, iodo and
methanesulfonyl. Preferably, the reactive moiety is a chloromethyl group.
Additionally, the polymer's backbone must be inert to both the carbodiimide
and reaction conditions under which the ultimate polymer bound coupling
reagents will be used.
Certain hydroxymethylated resins may be converted into
chloromethylated resins useful for the preparation of polymer supported
coupling reagents. Examples of these hydroxylated resins include the
4-hydroxymethylphenylacetamidomethyl resin (Pam Resin) and 4-
benzyloxybenzyl alcohol resin (Wang Resin) available from Advanced
Chemtech of Louisville, Kentucky, USA (see Advanced Chemtech 1993-1994
catalog, page 115). The hydroxymethyl groups of these resins may be
converted into the desired chloromethyl groups by any of a number of methods
well known to the skilled artisan.
Preferred resins are the chloromethylated styreneldivinylbenzene resins
because of their ready commercial availability. As the name suggests, these
resins are already chloromethylated and require no chemical modification prior
to use. These resins are commercially known as Merrifield's resins and are
available from Aldrich Chemical Company of Milwaukee, Wisconsin, USA (see
Aldrich 1994-1995 catalog, page 899). Methods for the preparation of PEPC
and its polymer supported forms are outlined in the following scheme.
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~NCO O
HZN~N
~H~H~N
O
~~CI
N~N-C -N
LG
Functionalized Resin
' where ~ = an inert polymer
and LG = CI, Br, I or OSOzCH~
N~N-C -N~
2o O/ CI
Such methods are described more fully in U.S. Patent Application Serial
No. 601019,790 filed June 14, 1996 which application is incorporated herein by
reference in its entirety. Briefly, PEPC is prepared by first reacting ethyl
isocyanate with 1-(3-aminopropyl)pyrrolidine. The resulting urea is treated
with 4-toluenesulfonyl chloride to provide PEPC . The polymer supported form
is prepared by reaction of PEPC with an appropriate resin under standard
conditions to give the desired reagent.
._. . . __.....~..~. ~. ~~ .. r
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The carboxylic acid coupling reactions employing these reagents are
performed at about ambient to about 45°C, for from about 3 to 120
hours.
' Typically, -the product may be isolated by washing the reaction with CHC13
and
concentrating the remaining organics under reduced pressure: As discussed
supra, isolation of products from reactions where a polymer bound reagent has
been used is greatly simplified, requiring only filtration of the reaction
mixture
and then concentration of the filtrate under reduced pressure.
Preparation of Cyclic Amino Compounds
Cyclic amino compounds 2 employed in reaction ( 1 ) above are generally
aminolactams, aminolactones, aminothiolactones and aminocycloalkyl
compounds which can be represented by the formula:
~C /Q
U
where U and s are as defined above, Q is preferably selected from the
group consisting of -O-) -S-, > NR'6, arid > CR"R'$ where each of R'6, R" and
R'8 are independently selected from the group consisting of hydrogen, alkyl,
substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl,
aryl,
heteroaryl and heterocyclic with the proviso that if Q is -O-, -S- or > NR'6,
then X is oxo or dihydro.
The aminolactams, aminolactones and aminothiolactones of the formula
above can be prepared by use or adaptation of known chemical syntheses which
syntheses are well described in the literature. See, e.g., Ogliaruso and
Wolfe,
Synthesis of Lactones and Lactams, Patai, et al. Editor, J. Wiley & Sons, New
' York, New York, USA, pp. 1085 et seq. (1993}.
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Specifically, 3-amino substituted lactams 13 with 5, 6 or 9 ring atoms
may be prepared by the direct cyclization of a suitable alpha, omega-diamino
acid ester-12 as shown in reaction (5) below:
O
R9
HZN-L -----~~- ~ Reaction (5)
NH-Pr NH-P r
1Z .L'~
IO
wherein L is a linking group (typically an alkylene group) of from 2-4 atoms,
Pr is a suitable protecting group such as t-butoxycarbonyl, carbobenzyloxy, or
the like and R'9 is an alkoxy or aryloxy group such as methoxy, ethoxy,
p-nitrophenoxy, N succinimidoxy, and the like. The reaction may be carried
out in a solvent such as water, methanol, ethanol, pyridine, and the like.
Such
reactions are exemplied by cyclization of a lysine ester to a caprolactam as
described by Ugi, et al., Tetrahedron, 52(35):11657-11664 (1996).
Alternatively, such a cyclization can also be conducted in the presence of
dehydrating agents such as alumina or silica to form lactams as described by
Blade-Font, Tetrahedron Lett. , 21:2443 (1980).
The preparation of aminolactams alkylated on the amino group of the
cyclic lactam is described by Freidinger, et al . , J. Org. Chem. , 47:104-109
(1982) and illustrated in reaction (6) below:
O
~..i H R 1. Reductive O
amination
L
Reaction (6}
H2N (~-~IFT O 2. Cycfization
NH-P r
r_ ~
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wherein L and R'6 are as defined above.
In reaction (6), reductive amination of 14 with aldehyde 15 and
subsequent ring closure by methods using, for example, EDC-provides for
aminolactam 16. The preparation of six-membered lactams using this general
procedure is described by Semple, et al., J. Med. Chem., 39:4531-4536 (1996).
The internal cyclization of an amide anion with a halide or equivalent
thereof can sometimes be used to particular advantage in the synthesis of
smaller ring lactams where the stereochemistry of the amino-lactam center is
available from the standard amino-acid pool. This approach is illustrated in
reaction (7) below:
SJ~te
N-R6 N-R6 Reaction (7)
BxHN ~ ~ gacHN
L o 0
1$
where R'6 is as defined above.
The approach of reaction (7) is presented by Semple, et al., supra.) and
Freidinger, et al., J. Org. Chem., 47:104-109 (1982) where a
dimethylsulfonium leaving group is generated from methyl iodide treatment of
an alkyl methyl sulfide 17 to provide for lactam 18. A similar approach using
a Mitsunobu reaction on an omega alcohol is found Holladay, et al., J. Org.
Chem., 56:3900-3905 (1991).
In another method, lactams 20 can be prepared from cyclic ketones 19
using either the well known Beckmann rearrangement (e.g., Donaruma, et al.,
Organic Reactions, 11:1-156 {1960)) or the well known Schmidt reaction
(Wolff, Organic Reactions, 3:307-336 (1946)) as shown in reaction (8) below:
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Beckmann rearrangement
r H
Schmidt reaction L-N - Reaction (8)
O
wherein L is as defined above.
Application of these two reactions leads to a wide variety of lactams
especially lactams having two hydrogen atoms on the carbon alpha to the lactam
carbonyl which lactams form a preferred group of lactams in the synthesis of
the compounds of formula I above. In these reactions, the L group can be
highly variable including, for example, alkylene, substituted alkylene and
hetero
i5 containing alkylene with the proviso that a heteroatom is not adjacent to
the
carbonyl group of compound 19. Additionally, the Beckmann rearrangement
can be applied to bicyclic ketones as described in Krow, et al. , J. Org.
Chem. ,
61:5574-5580 (1996).
The preparation of lactones can be similarly conducted using peracids in
a Baeyer-Villiger reaction on ketones. Alternatively) thiolactones can be
prepared by cyclization of an omega -SH group to a carboxylic acid and
thiolactams can be prepared by conversion of the oxo group to the thiooxo
group by PISS or by use of the commercially available Lawesson's Reagent,
Tetrahedron, 35:2433 (1979).
One recently reported route for lactam synthesis is a variation of the
Schmidt reaction through the use of an alkyl azide, either intermolecularly or
intramolecularly, through a tethered alkylazide function that attacks a ketone
under acidic conditions. Gracias, et al., J. Am. Chem. Soc., 117:8047-8048
(1995) describes the intermolecular version whereas Milligan, et al., J. Am.
r... ~ ..
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Cnem. Soc. , 117:10449-10459 (1995) describes the intramolecular-version.
One example of the intramolecular version is illustrated in reaction (9)
below:
- 5 O O
~N
N3
R1o Reaction (9)
R10
~"1 33'
where Rl° is exemplified by alkyl, substituted alkyl, alkoxy,
substituted alkoxy,
aryl, heteroaryl, cycloalkyl and heterocyclic.
In this reaction, ketone 21 is converted to an a-(w-alkyl)ketone 22 which
is cyclized to form bicyclic lactam 23. Such intramolecular reactions are
useful
in forming bicyclic lactams having 5-7 members and the lactam ring of 6-13
members. The use of hetero atoms at non-reactive sites in these rings is
feasible in preparing heterobicyclic lactams.
Still another recent approach to the synthesis of lactams is described by
Miller, et al . , J. Am. Chem. Soc. , 118 :9606-9614 ( 1996) and references
cited
and is illustrated in reaction (10) below:
y
l
pr, --.-~.~ .._____
H ~N~~6 HiV ~N. 6 HZN ~"~N. (10)
p Pr O R , ~ R
~4
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__ 6g __
where R6 and Pr are as defined above and Rl' is exemplified by halo, alkyl,
substituted alkyl, alkoxy, substituted alkoxy, aryl, heteroaryl, cycloaikyl
and
heterocyciic wherein the aryl, heteroaryl, cycloalkyl and heterocyclic group
is
optionally fused to the lactam ring structure.
Specifically, in reaction (10), lactam 26 is formed from an appropriate
unsaturated amide (e.g., ~ through a ruthenium or molybdenum complexes
catalysed olefin metathesis reaction to form unsaturated lactam 25 which can
be
used herein without further modification. However, the unsaturation in 25
permits a myriad of techniques such as hydroboration, Sharpless or Jacobsen
epoxidations, Sharpless dihydroxylations, Diels-Alder additions, dipolar
cycloaddition reactions and many more chemistries to provide for a wide range
of substituents on the lactam ring. Moreover, subsequent transformations of
the
formed substitution leads to other additional substituents (e.g. , mesylation
of an alcohol followed by nucleophilic substitution reactions). See, for
example, March, et al. Advanced Organic Chemistry, Reaction Mechanisms and
Structure, 2nd Edition, McGraw-Hill Book Company, New York, New York,
USA ( 1977) for a recitation of numerous such possible reactions. Saturated
amides used in this reaction are conventional with amide 24 being commercially
available.
Related chemistry to cyclize amides to form iactams is disclosed by
Colombo, et al . , Tetrahedron Lett. , 35 (23):4031-4034 ( 1994) and is
illustrated
in reaction (11) below:
I
AcHN ~N ~AcHN N Reaction (11}
~'j a
C02tBu C02tBu
~.. ~
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--69--
In this reaction, proline derivative 27 is cyclized via a tributyltin-radical
cyclization to provide for lactam 28.
Some of the lactams described above contain the requisite amino group
alpha to the lactam carbonyl whereas others did not. However, the introduction
of the required amino group can be achieved by any of several routes
delineated
below which merely catalogue several recent literature references for this
synthesis.
For example, in a first general synthetic procedure, azide or amine
displacement of a leaving group alpha to the carbonyl group of the lactam
leads
to the alpha-aminolactams. Such general synthetic procedures are exemplified
by the introduction of a halogen atom followed by displacement with
phthalimide anion or azide and subsequent conversion to the amine typically by
hydrogenation for the azide as described in Rogriguez, et al. , Tetrahedron,
52: 7727-7736 ( 1996), Parsons, et al. , Biochem. Biophys. Res. Comm. , 117
:108-
113 ( 1983) and Watthey, et al. , J. Med. Chem. , 28:1511-1516 ( 1985). One
particular method involves iodination and azide displacement on, for example,
benzyllactams as described by Armstrong, et al. , Tetrahedron Lett. , 35:3239
(1994) and by King, et a1. , J. Org. Chena. , 58:3384 ( 1993).
Another example of this first general procedure for the synthesis of
alpha-aminolactams from the corresponding lactam involves displacement of a
triflate group by an azido group as described by Hu, et al . , Tetrahedron
Lett. ,
36(21):3659-3662 (1995).
Still another example of this first general procedure uses a Mitsunobu
reaction of an alcohol and a nitrogen equivalent (either -NHZ or a phthalimido
group) in the presence of an azodicarboxylate and a triarylphosphine as
described in Wada, et al., Bull. Chem. Soc. Japan, 46:2833-2835 (1973) using
an open chain reagent.
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__ 7p __
Yet another example of this first general procedure involves reaction of
alpha-chlorolactams with anilines or alkyl amines in a neat mixture at
120°C to
provide for 2-(N-aryl or N-alkyl)lactams as described by Gaetzi, Chem. Abs. ,
66: 28690m.
In a second general synthetic procedure, reaction of an enolate with an
alkyl nitrite ester to prepare the alpha oxime followed by reduction yields
the
alpha-aminolactam compound. This general synthetic procedure is exemplified
by Wheeler, et al. , Organic Syntheses, Coll. Vol. VI, p. 840 which describes
the reaction of isoamyl nitrite with a ketone to prepare the desired oxime.
The
reduction of the oxime methyl ester (prepared from the oxime by reaction with
methyl iodide) is described in the J. Med. Chem. , 28(12):1886 (1985) and the
reduction of alpha-oximino caprolactams by Raney-nickel and palladium
catalysts is described by Brenner, et al., U.S. Patent No. 2,938,029.
In a third general synthetic procedure, direct reaction of an enolate with
an electrophilic nitrogen transfer agent can be used. The original reaction
employed toluenesulfonyl azide but was improved as described by Evans, et al.
,
J. Am. Chem. Soc. , I 12:4011-4030 { 1990) . Specifically, direct introduction
of
an azido group which can be reduced to the amine by hydrogenation is
described by Micouin, et al., Tetrahedron, 52:7719-7726 (1996). Likewise, the
use of triisopropylbenzenesulfonyl azide as the azide transferring agent for
reaction with an enolate is described by Evans, et al. , supra. The use of
triphenylphosphine to reduce the alpha-azidolactams to the corresponding
aminolactams in the benzodiazepine series is disclosed by Butcher, et al . ,
Tetrahedron Lett. , 37(37):6685-6688 (1996). Lastly, diazo transfer of beta-
diketones and subsequent reduction of the diazo group to the amino group is
exemplified by Hu, et al., Tetrahedron Lett. , 36(21):3659-3662 (1995) who
used Raney-nickel and hydrogen in acetic acid and acetic anhydride as the
solvent.
....... _~ .?.~._~.. _ r _
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__ 7I __
In a fourth general procedure, N substituted lactams are first converted
to the 3-alkoxycarbonyl derivatives by reaction with a dialkyl carbonate and a
' base such as sodium hydride. See, for example, M.L. Reupple, et al. , .1.
Am.
Chem. Soc. , 93:7021 et seq. ( 1971 ) The resulting esters serve as starting
materials for conversion to the 3-amino derivatives. This conversion is
achieved via the Curtius reaction as shown in reaction ( 12) below:
RI2 R 12 /R12
~-,~ L-N
L-N _
Reaction ( I2)
O ROC O- P~F-F'~ O
where Pr is as defined above and R'z is typically hydrogen, an alkyl or an
aryl
group. ,
The Curtius reaction is described by P.A.S. Smith, Organic Reactions,
3 : 337-449 ( 1946) . Depending on the reaction conditions chosen, Pr = H or a
protecting group such as Boc. For example, when R = H, treatment of the
acid with diphenylphosphoryl azide in the presence of t-butanol provides the
product wherein Pr = Boc.
The alpha-aminolactams employed as the cyclic amino compounds 2 in
reaction {1) above include ring N-substituted lactams in addition to ring N-H
lactams. Some methods for preparing ring N-substituted lactams have been
described above. More generally, however, the preparation of these
compounds range from the direct introduction of the substituent after lactam
formation to essentially introduction before lactam formation. The former
methods typically employ a base and an primary alkyl halide although it is
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contemplated that a secondary alkyl halide can also be employed ajthough
yields
may suffer.
Accordingly, a first general method for preparing N-substituted lactams
is achieved via reaction of the lactam with base and alkyl halide (or
acrylates in
some cases). This reaction is quite well known and bases such as sodamide,
sodium hydride, LDA, LiHMDS in appropriate solvents such as THF, DMF,
etc. are employed provided that the selected base is compatible with the
solvent. See for example: K. Orito, et al., Tetrahedron, 36:1017-1021 (1980)
and J. E. Semple, et al . , J. Med. Chem. , 39: 4531-4536 ( 1996} (use of
LiHMDS
. with either R-X or acrylates as electrophiles).
A second general method employs reductive amination on an amino
function which is then cyclized to an appropriate ester or other carbonyl
function.
A third general method achieves production of the N-substitution during
lactam formation. Literature citations report such production from either
photolytic or thermal rearrangement of oxaziridines, particularly of N-aryl
compounds. See, for example, Krimm, Chem. Ber., 91:1057 (1958) and Suda,
et al., J. Chem. Soc. Chem Comm., 949-950, (1994). Also, the use of methyl
hydroxylamine for the formation of nitrones and their rearrangement to the N-
methyl derivatives is reported by Barton, et al. , J. Chem. Soc. , 1764-1767
( 1975). Additionally, the use of the oxaziridine process in chiral synthesis
has
been reported by Kitagawa, et al . , J. Am. Chem. Soc. , 117 : 5169-5178 (
1975 ) .
A more direct route to obtain N-phenyl substituted Iactams from the
corresponding NH lactams through the use of t-butyltetramethylguanidine and
triphenylbismuth dichloride is disclosed by Akhatar, et al . , J. Org. Chem. ,
, 55:5222-5225 (1990) as shown in reaction (13) below.
..____....~...u .... . . ~
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__ ~3 __
O O
O ~ O Reaction ( 13)
O N ~~ O N
H
Given that numerous methods are available to introduce an alpha-amino
group onto a lactam (or lactone) ring, the following lactams (and appropriate
corresponding lactones) are contemplated for use in the synthesis of compounds
of formula I above. Similar alcohol functions at the carbonyl position are
derivative of either amine ring opening of cyclic epoxides, ring opening of
aziridines, displacement of appropriate halides with amine or alcohol
nucleophiles, or most likely reduction of appropriate ketones. These ketones
are also of interest to the present invention.
Monocyclic lactams as described by Nedenskov, et al., Acta Chem.
Scand. , 12:1405-1410 ( 1958) and represented by the formula:
O
NH R1
2
R
where R' and R2 are exemplified by alkyl) aryl or alkenyl (e.g., allyl).
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Monocyclic lactams containing a second nitrogen ring atom as described
by Sakakida, et al . , Bull. Chem. Soc. Japan, 44:478-480 ( 1971 ) and
represented by the formula:
O
NH
N
r
R
where R is exemplified by CH3- or PhCH2-.
15
Monocyclic lactams having hydroxyl substitution on the ring as
described by Hu, et al. , Tetrahedron Lett. , 36(21):3659-3662 (1995) and
represented by the formula:
HO
l '
H2N ~N~
0 R
where R is exemplified by benzyl (includes both the cis and traps hydroxy
lactams).
The direct preparation N substituted lactams of 5-8 members from the
corresponding ketones is described by Hoffman, et al . , Tet. Lett. , 30:4207-
4210
( 1989) . These lactams are represented by the formula:
0 O
---..~ N, R n = 1 _ 4
(CHZ)n (CH2)n
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wherein R is alkyl, alkenyl, alkynyl, cycioalkyl, or benzyl.
' N-methoxylactams prepared from cyclohexanone and dimethoxyamine
are described by Vedej s, et al . , Tet. Lett. , 33 : 3261-3264 ( 1992) .
These
structures are represented by the formula:
p
O
~CH3
Substituted 3-aminoazetidinone derivatives prepared by a variety of
routes including those described by van der Steen) et al. , Tetrahedron, 47,
7503-7524 ( 1991 )56, Hart, et al . , Chem Rev. , 89 :1447-1465 ( 1989) and
references cited therein are represented by the formula:
H2N R1
N
O .R2
where R1 and RZ are independently selected from alkyl, substituted alkyl,
alkenyl, substituted alkenyl, aryl, heteroaryl, heterocyclic or are fused to
form
a cyclic group.
Ring substituted lactams are described by Lowe, et al., Bioorg. Med.
Chem. Lett., 4:2877-2882 (1994) and are represented by the formula:
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R3
R _
H2N N,
RI
O
wherein Rz and R3 are exemplified by aryl and substituted aryl and R1 is
exemplified by alkyl or hydrogen.
The synthesis of substituted 3-aminopyrrolidones from alpha-
bromoketones is described by McKennis, Jr. , et al. , J. Org. Chem. , 28:383-
387
(1963). These compounds are represented by the formula:
NH2
O
Rl~Br ~ Ri N O
R 2
where Rl is aryl or heteroaryl and RZ corresponds to any substituent for which
the corresponding amine RZ-NHZ exists.
Additional references for the synthesis of alpha aminolactams are as
follows:
1. Shirota, et al. , J. Med. Chem. , 20:1623-1627 ( 1977) which describes
the synthesis of a compound of the formula:
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_ CHs
CH3
H2N ~ N,
O H
2. Overberger, et al., J. Am. Chem. Soc. , 85:3431 (1963) which
describes the preparation of optically active (3-methylcaprolactam of the
formula:
O N
H
3. Herschmann, Helv. Chim. Acta, 32:2537 (1949) describes the
synthesis of a disubstituted caprolactam from the Beckmann rearrangement of
menthone which is represented by the formula:
O
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__ ~g __
4. Overberger, et al., Macromolecules, 1:1 (1968) describes the
synthesis of eight-membered lactams from 3-methylcycloheptanone as shown
below:
S
O N + O
H N
O H
S. The synthesis of benzolactams (benzazepinones) has been reported by
Busacca, et al., Tet. Lett. , 33:165-168 (1992):
O
O JH
by Croisier, et al., U.S. Patent No. 4,080,449:
O
.....~r-._.~.,... .~
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_-
and by J. A. Robl, et al. , Tetrahedron Lett. , 3b( 10) :1593-1596 ( 1993) who
employed an internal Friedel-Crafts like cyclization to prepare the tricyclic
' benzyllactam's shown below where Pht is the phthalimido protecting group:
ELO OEt
--
Pht=N N Pit=N HZN
O C02Et C02Et C02Et
l0
IO
Another tricyclic lactam series is disclosed by Flynn, et al.) J. Med.
Chem. , 36:2420-2423 (1993) and references cited therein.
6. Orito, et al.) Tetrahedron, 36:1017-1021 (1980) discloses phenyl
substituted benzazepinones represented by the formula:
O
N-R
wherein R = H or CH3-;
Kawase, et al. , J. Org. Chem. , 54: 3394-3403 ( 1989) discloses a N-
methoxy benzazepinone represented by the formula:
~l
_ 30 ~ N
O
O,
CH3
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__ g0 __
7. Lowe, et al. , J. Med. Chem. , 37:3789-3811 ( 1994) describes several
synthetic pathways to substituted benzazepinones of the formula:
R~
NH2
N
' O
R2
where R' is substituted aryl or cyclohexyl) X is a suitable substituent and Rz
can be H or alkyl. The syntheses described in Lowe are, however, adaptable to
form numerous R' substituents.
8. Robl, et al., Bioorg. Med. Chem. Lett., 4:1789-1794 (1994) and
references cited therein as well as Skiles, et al . , Bioorg. Med. Chem. Lett.
,
3:773-778 (1993) disclose benzofused lactams which contain additional
heteroatoms in the lactam ring. These compounds are represented by the
formula:
X R
/ NH2
N
R1 O
where X is O and R'- = H or CH3 or X = S and RZ = H. In either case, R' _
H or alkyl. Also, in Skiles, the thio group of the thiolactam can be oxided to
the S02 group. These structures are also presented from Beckmann
rearrangement in Grunewald, et al . , J. Med. Chem. , 39( 18) : 3539 ( 1996) .
~, r
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__ g 1 __
9. Also syntheses for the benzoheterolactam series is presented in
Thomas ) et al. , J. Chem. Soc. , Perkin II, 747 ( 1986) which could lead to
compounds of the formula:
IVH2 _
' S ~X
O N -N
R
X = O, H2
R ~ COZR
where X is O or H2 and R is COZR.
10. Further examples of benzazepinones are found in Warshawsky, et
al. , Bioorg. Med. Chem. Lett. , 6: 957-962 ( 1996) which discloses compounds
of
the formula:
N-R
H2N
The synthesis can be generalized to produce R = alkyl or aryl.
1 I . Ben-Ishai, et al. , Tetrahedron, 43:439-450 ( 1987) describes
syntheses which could lead to several benzolactams of the formula:
~(CH2)n
~N-R
X O
H2N
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__ g2 __
wherein n = 0) l, or 2 and R= -CH3, PhCH,- and H.
12. van Niel et al., Bioorg. Med. Chem. Len., S:i4Z1-1426 (1995)
reports the synthesis of compounds of the fotTnulas:
Hs N O H3C O
j w ~ N
NH2 ~ NHBoc
i
to
x o
wherein X is -OH, -NHZ or -NR6R6 where R6 is as defined above. The
reported ketone is a versatile synthetic intermediate which can be modified by
conventional methods such as reductive amination, reduction, etc.
13. Kawase, et ai.) J. Org. Chem., 54:3394-3403 (1989) describes a
synthetic method for the preparation of:
~ N H2
i~
N 0
OCH3
In addition to the above, saturated bicyclic alpha-aminolactams are also
contemplated for use in the synthesis of compounds of formula I. Such
saturated bicyclic alpha-aminolactams are well known in the art. For example,
Edwards, et al. , Can. J. Chem. , 49:1648-1658 ( 1971 ) describes several
syntheses of bicyclic lactams of the formula:
. _.__ _. r
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__ g3 __
a o
Similarly ) Milligan, et al. , J. Am. Chem. Soc. , 117:10449-10459 ( 1995)
and references cited therein report the synthesis of lactams of the formula:
p R2 O
/ Z N N
N A B
fll
R
wherein R' and RZ are H or -CH3, ring A can have from 6-13 members and
ring B can have from 5-7 members. R can be alkyl, aryl, cycloalkyl and the
like.
The introduction of a heteroatom into the saturated cyclic structure fused
to the lactam ring is disclosed by Curran et al., Tet. Lett., 36:191-194
(1995),
who describe a synthetic method which can be used to obtain a lactam of the
formula:
NH
N
p NH2
by Slusarchyk, et al., Bioorg. Med. Chem. Lett. , 5:753-758 (1995), who
describe syntheses which could lead to a lactam of the formula:
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__ g4 __
~S
H2N
O
and by Wyvratt, et al., Eur. Pat. Appl. 61187 (1982), who describe a lactam of
the formula:
--S
NJ
H2N
O
IS
Lactams having further heteroatom(s) in the cyclic lactam structure (in
addition to the nitrogen of the amido group of the lactam) are described by
Cornille, et al., ,l. Am. Chem. Soc., 117:909-917 (1995), who describe lactams
of the formula:
O
N
H2N
O
and J. Kolc, Coll. Czech. Chem. Comm. , 34:630 ( 1969), who describes lysines
suitable for cyclization to lactams which have a hetero lactam ring atom as
shown by the formula:
H2N X
-,". ~ X
O ___
O
N
H
r~. r
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__ gs __
where X = O, S and NR where R is, for example, alkyl, substituted alkyl ) aryl
heteroaryl, heterocyclic) and the like.
Similarly, each of Dickerman, et al., J. Org. Chem., 14:530 (1949)86,
Dickerman, et al . , J. Org. Chem. , 20:206 ( 1955), and Dickerman, et al. ,
J.
Org. Chem., 19:1855 (1954) used the Schmidt and Beckmann reactions on
substituted 4-piperidones to provide for lactams of the formula:
R\-~ N.R R\.., N.R
J..
O N
H
where R is acyl, alkyl, substituted alkyl, aryl, heteroaryl or heterocyclic
IS provided that R is not an acid labile group such as t-Boc; and R' is
hydrogen,
alkyl, substituted alkyl, alkoxy, substituted alkoxy, aryl, aryloxy,
heteroaryl,
heteroaryloxy, heterocyclic, heterocyclicoxy, halo, cyano, vitro,
trihalomethyl,
and the like.
An internal cyclization of appropriate ethylenediamine amides onto a
ketone or aldehyde is described by Hoffman, et al. , J. Org. Chem. , 27:3565
(1962) as follows:
R
O Z R
HN N
/_
O 'w
N O
H N
H
R = Methyl, Pheny
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__ g6 __
10
Ring expansion methodology based on beta lactams to provide for larger
ring lactams containing an aza group has twice been reported in Wasserman, et
al. , J. Am. Chem. Soc. , 103 :461-2 ( 1981 ) and in Crombie, et al . ,
Tetrahedron
Lett., 27(42):5151-5154 (1986). -
Dieckmann methodology has been used to prepare aza caprolactams
from unsymmetrical amines such as shown below by Yokoo, et al . , Bull, Chem.
Soc. Jap., 29:631 (1956).
R
N. R
N
C02Et -----
C02Et O N
H
where R is as defined in this reference. The dislosure of Yokoo, et al. can be
extended to cover R being alkyl, substituted alkyl, aryl, alkoxy, substituted
alkoxy, heteroaryl, cycloalkyl, heterocyclic, aikenyl, substituted alkenyl,
and
the Like.
The synthesis of various members of the oxalactam series has been
reported by Burkhoider, et al. , Bioorg. Med. Chem. Lett. , 2:231 ( 1993) and
references cited therein which oxalactams are represented by the formula:
'RHN p
O N
R
~ ~
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__ g7 __
where 'R is as defined in the reference and R can be alkyl, substituted alkyl,
aryl, alkoxy, substituted alkoxy) heteroaryl) cycloalkyl) heterocyclic,
alkenyl,
substituted alkenyl, and the like.
The synthesis of thialactams {generally oxalactams can be made by the
same methodology} has been reported by Freidinger) et al. , J. Org. Chem. ,
47:104-109 (1982), who prepared thialactams of the formula:
S1
H2N N~R
O
This reference provides a series of procedures having broad application for
synthesis of lactams permitting R in the above formula to be derived from any
amine (alkyl, aryl, heteroaryl, etc. ) with the restriction being that the R-
group
does not contain any functional groups reactive with formaldehyde (e. g. ,
primary and secondary amines). The general synthetic scheme provided by
Freidlinger, et al. is:
H H
~- N
O S ~J- coupling O S
O agent ~ _ O
~N + R- NHZ ~ N
OH ~ NHR
0 0 O 0
S S
paraformaidehyde 0 ~ N-methylhydrazine
p-TosOH ~ ~ 'N O R H2N N~R
Ci2CHCHCrz ~~O -O
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The coupling agent is any standard reagent used in the formation of typical
peptide or-amide bonds, for example, carbodiimide reagents. See) also,
Karanewsky, U.S. Patent No. 4,460,579 and Kametani, et aI., Heterocycles,
9:831-840 (1978). -
S
The Friedinger procedure can be extended to afford disubstituted
thialactams of the following structure:
R2
HZN N~ R1
0
In this procedure, the thiolactam ring is prepared as follows:
S R2
R 2 coupling
N SH .+ ~ agent N N..R1
OH N' Rl ~ ~ 0
In practical terms, RZ will be limited to aryl and heteroaryl groups and
sterically hindered alkyl groups such as t-butyl. R1 can be highly variable
and
is limited only by subsequent reaction steps.
Still further is the Kametani procedure which provides for lactams as
follows:
_~~,-. .."~_.~.". ...._ . _.., . .... _ . ?
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O O S R2
H R 2 P-TosOH
SH benzene N N. R 1
N + O
NHR~ ~ ~ O
O O O
In principle, the Kametani procedure allows for a wide selection of R' and R'-
groups limited primarily by stability to the reaction conditions.
See> for example, Yanganasawa, et al., J. Med. Chem., 30:1984-1991
( 1987} and J. Das et al . , Biorg. Med. Chem. Lett. , 4:2193-2198 ( 1994)
which
describes general methods for the synthesis of isomeric 7-membered thialactams
of the following structure:
R2
S S
~ R2
H2N ll N 1 H2N N
~R1
O R O
The first synthetic route is:
RZ
~ SH 2 1. N-meth I-
R Y S
_ morphoiine
- BocHN OH ~' ~"'~ -----~ N H2
N02 2. Reduction BocHN OH
O
2 O
R
~ . Cyclization S
2. Selective
aliKyi;ation H2N N 1
O ~R
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RZ can be highly variable (e. g. , alkyl, substituted alkyl, aryl, heteroaryl,
heterocyclic and the like) since a number of well documented routes exist for
the synthesis of nitroethylene derivatives from aldehydes and nitromethane
(Henry reaction) followed by dehydration. R' is limited to groups that can
undergo alkylation reactions.
O ~NHBoc
O
SH R 2 NHBoc ~ S R2
,N + ~ N
~ OCHPh2 OMs ~ OCHPh2
O O O O
1. Protecting group
~R2
removal
H2N ~ N 1
IS 2. Cyclization 'R
3. Selective alkylation (R~) O
4. Mettiylhydrazine
In this procedure, RZ can be highly variable. The starting component required
to introduce R2 can be readily derived by the reduction of any known alpha-
BOC-amino acid to the alcohol derivative followed by formation of the
mesylate.
As noted above, the primary approaches to the preparation of lactams is
the Beckmann/Schmidt ring expansion reaction using either inter- or
intramolecular approaches serves to prepare lactams of various ring sizes. The
intramolecular approach generates bicyclic materials with the lactam nitrogen
incorporated into the ring fusion. Additional approaches set forth above are
at
the base of the methodology are internal cyclization of omega-amino
acids/esters where the construction of the substituent pattern takes place
prior to
cyclization, and internal cyclization of an electrophilic center onto a
....~____-.~,.._...._ ,
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nucleophilic functional group as in the Friedel Crafts type cyclization at the
center of the Ben-Ishal procedure for making benzazepinones. This latter
procedure is-applicable to a wide variety of heteroaromatics as well as
benzenoid rings, and may also be applied to non-aromatic double or triple
bonds to generate a wide array of substituents or ring fusions.
Deoxygenation of the lactam by reagents such as diborane, LiAlH4, and
the like leads to azaheterocycles (=X is dihydro).
Similarly, for U = H, OH, such compounds can be prepared by
epoxidation of cycloalkenyl groups followed by oxirane opening by, e.g. ,
ammonia. After formation of compounds of formula I, =U being H, OH can
be oxidized to provide for cycloalkylones (=X being oxo).
Additional ly ) the 5 , 7-dihydro-6H-diben [b, d] azepin-6-one derivatives
employed in this invention can be prepared using conventional procedures and
reagents. For example, an appropriately substituted N-ten-Boc-2-amino-2'-
methylbiphenyl compound can be cyclized to form the corresponding 5,7-
dihydro-6H-diben[b,d]azepin-6-one derivative by first treating the biphenyl
compound with about 2.1 to about 2.5 equivalents of a strong base, such as sec-
butyl lithium. This reaction is typically conducted at a temperature ranging
from about -80°C to about -60°C in an inert diluent such as THF.
The
resulting dianion is then treated with dry carbon dioxide at a temperature of
about -78°C to afford the 5,7-dihydro-6H-diben[b,d]azepin-6-one. This
procedure is described further in R. D. Clark et al. , Tetrahedron, 49(7),
1351-
1356 ( 1993) and references cited therein.
After forming the 5,7-dihydro-6H-diben[b,d]azepin-6-one, the amide
nitrogen can be readily alkylated by first treating the dibenazepinone with
about
1.1 to about 1.5 equivalents of a strong base, such as sodium hydride, in an
inert diluent, such as DMF. This reaction is typically conducted at a
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temperature ranging from about - I 0 ° C to about 80 ° C for
about 0. 5~ to about b
hours. The resulting anion is then contacted with an excess, preferably about
1.1 to about 3.0 equivalents, of an alkyl halide, typically an alkyl chloride,
bromide or iodide. Generally, this reaction is conducted at a temperature of
about 0°C to about 100°C for about 1 to about 48 hours.
An amino group can then be introduced at the 5-position of the 7-alkyl-
5,7-dihydro-6H-diben[b,d]azepin-6-one using conventional procedures and
reagents. For example, treatment of 7-methyl-5,7-dihydro-6H-
diben[b,d]azepin-6-one with an excess of butyl nitrite in the presence of a
strong base, such as potassium 1,1,1,3,3,3-hexamethyldisilazane (KHMDS),
affords 5-oximo-7-methyl-5,7-dihydro-6H-diben[b,d]azepin-6-one. Subsequent
reduction of the oximo group by hydrogenation in the presence of a catalyst,
such as palladium on carbon, then provides 5-amino-7-methyl-5,7-dihydro-6H-
diben[b,d]azepin-6-one. Other conventional amination procedures, such as
azide transfer followed by reduction of the azido group, may also be employed.
Similarly, various benzodiazepine derivatives suitable for use in this
invention can be prepared using conventional procedures and reagents. For
example, a 2-aminobenzophenone can be readily coupled to a-(isopropylthio)-
N-(benzyloxycarbonyl)glycine by first forming the acid chloride of the glycine
derivative with oxayl chloride, and then coupling the acid chloride with the 2-
aminobenzophenone in the presence of a base, such as 4-methylmorpholine, to
afford the 2-[a-(isopropylthio)-N-(benzyloxycarbonyl)glycinyl]-
aminobenzophenone. Treatment of this compound with ammonia gas in the
presence of an excess, preferably about 1.1 to about 1.5 equivalents, of
mercury (II) chloride then affords the 2-[N-(a-amino)-N'-(benzyloxycarbonyl)-
glycinyl]aminobenzophenone. This intermediate can then be readily cyclized by
treatment with glacial acetic acid and ammonium acetate to provide the 3-
(benzyloxycarbonyl)amino-2,3-dihydro-5-phenyl-IH-1,4-benzodiazepin-2-onel.
...._.. ."._...,.~.....u,..~..~._.-..... . .. .. ?,.. . .. , ..
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( -_ g3 __
Subsequent removal of the Cbz group affords the 3-amino-2,3-dihydro-5-
phenyl-1 H-1,4-benzodiazepin-2-one .
Alternatively, 2,3-dihydro-S-phenyl-1H-1,4-benzodiazepin-2-ones can be
readily aminated at the 3-position using conventional azide transfer reactions
followed by reduction of the resulting azido group to form the corresponding
amino group. The conditions for these and related reactions are described in
the examples set forth below. Additionally, 2,3-dihydro-5-phenyl-1H-1,4-
benzodiazepin-2-ones are readily alkylated at the 1-position using
conventional
procedures and reagents. For example, this reaction is typically conducted by
first treating the benzodiazepinone with about 1.1 to about 1.5 equivalents of
a
base, such as sodium hydride, potassium ten-butoxide, potassium 1,1,1,3,3,3-
hexamethyldisilazane, cesium carbonate, in an inert diluent, such as DMF.
This reaction is typically conducted at a temperature ranging from about -78
°C
to about 80°C for about 0.5 to about 6 hours. The resulting anion is
then
contacted with an excess, preferably about 1.1 to about 3.0 equivalents, of an
alkyl halide, typically an alkyl chloride, bromide or iodide. Generally, this
reaction is conducted at a temperature of about 0 ° C to about 100
° C for about 1
to about 48 hours.
Additionally, the 3-amino-2,4-dioxo-2,3,4,5-tetrahydro-1H-1,5-
benzodiazepines employed in this invention are typically prepared by first
coupling malonic acid with a 1,2-phenylenediamine. Conditions for this
reaction are well known in the art and are described, for example, in PCT
Application WO 96-US8400 960603. Subsequent alkylation and amination
using conventional procedures and reagents affords various 3-amino-1,5-
bis(alkyl)-2,4-dioxo-2,3,4,5-tetrahydro-1H-1,5-benzodiazepines. Such
procedures are described in further detail in the example set forth below.
- 30 Accordingly, a vast number of lactams, lactones and thiolactones are
available by art recognized procedures. Similarly, the art is replete with
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examples of aminocycloalkyl compounds for use in the synthesis of-compounds
of formula I above.
In the synthesis of compounds of formula I using the synthetic methods
described above, the starting materials can contain a chiral center (e. g. ,
alanine)
and, when a racemic starting material is employed, the resulting product is a
mixture of R,S enantiomers. Alternatively, a chiral isomer of the starting
material can be employed and, if the reaction protocol employed does not
racemize this starting material, a chiral product is obtained. Such reaction
protocols can involve inversion of the chiral center during synthesis.
Accordingly, unless otherwise indicated, the products of this invention
are a mixture of diastereomers or R,S enantiomers. Preferably, however, when
a chiral product is desired, the chiral product corresponds to the L-amino
acid
derivative. Alternatively, chiral products can be obtained via purification
techniques which separates diastereomers or enantiomers to provide for one or
the other stereoisomer. Such techniques are well known in the art.
Pharmaceutical Formulations
When employed as pharmaceuticals, the compounds of formula I are
usually administered in the form of pharmaceutical compositions. These
compounds can be administered by a variety of routes including oral, rectal,
transdermal, subcutaneous, intravenous, intramuscular, and intranasal. These
compounds are effective as both injectable and oral compositions. Such
compositions are prepared in a manner well known in the pharmaceutical art
and comprise at least one active compound.
This invention also includes pharmaceutical compositions which contain,
as the active ingredient, one or more of the compounds of formula I above
associated with pharmaceutically acceptable carriers. In making the
compositions of this invention, the active ingredient is usually mixed with an
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excipient, diluted by an excipient or enclosed within such a carrier which can
be in the form of a capsule, sachet, paper or other container. When the
excipient sefves as a diluent, it can be a solid, semi-solid, or liquid
material,
which acts as a vehicle, carrier or medium for the active ingredient. Thus,
the
compositions can be in the form of tablets, pills, powders, lozenges, sachets,
cachets, elixirs, suspensions, emulsions, solutions, syrups, aerosols (as a
solid
or in a liquid medium), ointments containing, for example, up to 10% by
weight of the active compound, soft and hard gelatin capsules, suppositories,
sterile injectable solutions, and sterile packaged powders.
In preparing a formulation, it may be necessary to mill the active
compound to provide the appropriate particle size prior to combining with the
other ingredients. If the active compound is substantially insoluble, it
ordinarily is milled to a particle size of less than 200 mesh. If the active
compound is substantially water soluble, the particle size is normally
adjusted
by milling to provide a substantially uniform distribution in the formulation,
e.g. about 40 mesh.
Some examples of suitable excipients include lactose, dextrose, sucrose,
sorbitol, mannitol, starches, gum acacia, calcium phosphate, alginates,
tragacanth, gelatin, calcium silicate, microcrystalline cellulose,
polyvinylpyrrolidone, cellulose, sterile water, syrup, and methyl cellulose.
The
formulations can additionally include: lubricating agents such as talc,
magnesium stearate, and mineral oil; wetting agents; emulsifying and
suspending agents; preserving agents such as methyl- and propylhydroxy-
benzoates; sweetening agents; and flavoring agents. The compositions of the
. invention can be formulated so as to provide quick, sustained or delayed
release
of the active ingredient after administration to the patient by employing
procedures known in the art.
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The compositions are preferably formulated in a unit dosage -form; each
dosage containing from about 5 to about 100 mg, more usually about 10 to
about 30 mg; of the active ingredient. The term "unit dosage forms" refers to
physically discrete units suitable as unitary dosages for human subjects and
other mammals, each unit containing a predetermined quantity of active
material calculated to produce the desired therapeutic effect, in association
with
a suitable pharmaceutical excipient. Preferably, the compound of formula I
above is employed at no more than about 20 weight percent of the
pharmaceutical composition, more preferably no more than about Z 5 weight
percent, with the balance being pharmaceutically inert carrier(s).
The active compound is effective over a wide dosage range and is
generally administered in a pharmaceutically effective amount. It, will be
understood, however, that the amount of the compound actually administered
will be determined by a physician, in the light of the relevant circumstances,
including the condition to be treated, the chosen route of administration, the
actual compound administered, the age, weight, and response of the individual
patient, the severity of the patient's symptoms, and the like.
For preparing solid compositions such as tablets, the principal active
ingredient is mixed with a pharmaceutical excipient to form a solid
preformulation composition containing a homogeneous mixture of a compound
of the present invention. When referring to these preformulation compositions
as homogeneous, it is meant that the active ingredient is dispersed evenly
throughout the composition so that the composition may be readily subdivided
into equally effective unit dosage forms such as tablets, pills and capsules.
This
solid preformulation is then subdivided into unit dosage forms of the type
described above containing from, for example, 0.1 to about 500 mg of the
active ingredient of the present invention.
...._... ..,~~"~"~"m"",..:.......,...,..,._...." ... . Y
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__ 9
The tablets or pills of the present invention may be coated or otherwise
compounded to provide a dosage form affording the advantage of prolonged
action. For-example, the tablet or pill can comprise an inner dosage and an
outer dosage component, the latter being in the form of an envelope over the
former. The two components can separated by enteric layer which serves to
resist disintegration in the stomach and permit the inner component to pass
intact into the duodenum or to be delayed in release. A variety of materials
can
be used for such enteric layers or coatings, such materials including a number
of polymeric acids and mixtures of polymeric acids with such materials as
shellac, cetyl alcohol, and cellulose acetate.
The liquid forms in which the novel compositions of the present
invention may be incorporated for administration orally or by injection
include
aqueous solutions suitably flavored syrups, aqueous or oil suspensions, and
flavored emulsions with edible oils such as cottonseed oil, sesame oil,
coconut
oil, or peanut oil, as well as elixirs and similar pharmaceutical vehicles.
Compositions for inhalation or insufflation include solutions and
suspensions in pharmaceutically acceptable, aqueous or organic solvents, or
mixtures thereof, and powders. The liquid or solid compositions may contain
suitable pharmaceutically acceptable excipients as described supra. Preferably
the compositions are administered by the oral or nasal respiratory route for
local or systemic effect. Compositions in preferably pharmaceutically
acceptable solvents may be nebulized by use of inert gases. Nebulized
solutions
may be breathed directly from the nebulizing device or the nebulizing device
may be attached to a face masks tent, or intermittent positive pressure
breathing
machine. Solution, suspension, or powder compositions may be administered,
preferably orally or nasally, from devices which deliver the formulation in an
appropriate manner.
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The following formulation examples illustrate the pharmaceutical
compositions of the present invention.
Formulation Example I
Hard gelatin capsules containing the following ingredients are prepared:
Quantity
In reg diem (mg/capsule)
Active Ingredient 30.0
Starch 305 .0
Magnesium stearate 5.0
The above ingredients are mixed and filled into hard gelatin capsules in
340 mg quantities.
Formulation Example 2
A tablet formula is prepared using the ingredients below:
Quantity
~20 Ingredient (mg/tablet)
Active Ingredient 25.0
Cellulose, microcrystalline 200.0
Colloidal silicon dioxide 10.0
Stearic acid 5.0
The components are blended and compressed to form tablets, each
weighing 240 mg.
Formulation Example 3
A dry powder inhaler formulation is prepared containing the following
components:
Ingredient Weight
Active Ingredient 5
Lactose 95
.._... "-".~..r"._ .._. ~
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The active ingredient is mixed with the lactose and the mixture is added
to a dry powder inhaling appliance.
Formulation Example 4 -
- 5 Tablets, each containing 30 mg of active ingredient, are prepared as
follows:
Quantity
In redient (~Itablet)
Active Ingredient 30.0 mg
Starch 45.0 mg
Microcrystalline cellulose 35.0 mg
Polyvinylpyrrolidone
(as 20 % solution in sterile water) 4.0 mg
Sodium carboxymethyl starch 4.5 mg
Magnesium stearate 0.5 mg
Talc 1.0 m~
Total I20 mg
The active ingredient, starch and cellulose are passed through a No. 20
mesh U. S . sieve and mixed thoroughly. The solution of polyvinyl-pyrrolidone
is mixed with the resultant powders, which are then passed through a 16 mesh
U.S. sieve. The granules so produced are dried at 50° to
60°C and passed
through a 16 mesh U. S. sieve. The sodium carboxymethyl starch, magnesium
stearate, and talc, previously passed through a No. 30 mesh U. S . sieve, are
then added to the granules which, after mixing, are compressed on a tablet
machine to yield tablets each weighing 150 mg.
Formulation Example 5
Capsules, each containing 40 mg of medicament are made as follows:
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Quantity
Ingredient (mg/capsule~
Active Ingredient 40.0 mg
Starch 109.0 mg
Magnesium stearate l,p m~
Total 150.0 mg
The active ingredient, starch, and magnesium stearate are blended, passed
through a No. 20 mesh U.S. sieve, and filled into hard gelatin capsules in 150
mg quantities.
Formulation Example 6
Suppositories, each containing 25 mg of active ingredient are made as
follows:
In reg diem Amount
Active Ingredient 25 mg
Saturated fatty acid glycerides to 2,000 mg
The active ingredient is passed through a No. 60 mesh U.S. sieve and
suspended in the saturated fatty acid glycerides previously melted using the
minimum heat necessary. The mixture is then poured into a suppository mold
of nominal 2.0 g capacity and allowed to cool.
Formulation Example 7
Suspensions, each containing 50 mg of medicament per 5.0 ml dose are
made as follows:
_.._...b~ _. .. . ,
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In redient Amount
Active Ingredient 50.0 mg
Xanthan gum 4.0 mg_
Sodium carboxymethyl cellulose (11 %o)
Microcrystalline cellulose (89%) 50.0 mg
Sucrose 1.75 g
Sodium benzoate 10.0 mg
Flavor and Color q. v.
Purified water to 5.0 ml
The active ingredient, sucrose and xanthan gum are blended, passed
through a No. 10 mesh U.S. sieve, and then mixed with a previously made
solution of the microcrystalline cellulose and sodium carboxymethyl cellulose
in
water. The sodium benzoate, flavor, and color are diluted with some of the
water and added with stirring. Sufficient water is then added to produce the
required volume.
Formulation Example 8
Quantity
In reg dient (m/capsule)
Active Ingredient 15.0 mg
Starch 407.0 mg
Magnesium stearate 3.0 m~
Total 425.0 mg
The active ingredient, starch, and magnesium stearate are blended, passed
through a No. 20 mesh U.S. sieve, and filled into hard gelatin capsules in 560
mg quantities.
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Formulation Exam In a 9
A subcutaneous formulation may be prepared as follows:
Ingredient uantit -
Active Ingredient 1.0 mg
corn oil 1 ml
(Depending on the solubility of the active ingredient in corn oil, up to
about 5.0 mg or more of the active ingredient may be employed in this
formulation, if desired).
Formulation Example 10
A topical formulation may be prepared as follows:
Ingredient uantit
Active Ingredient 1-10 g
Emulsifying Wax 30 g
Liquid Paraffin 20 g
White Soft Paraffin to 100 g
The white soft paraffin is heated until molten. The liquid paraffin and
emulsifying wax are incorporated and stirred until dissolved. The active
ingredient is added and stirring is continued until dispersed. The mixture is
then cooled until solid.
Another preferred formulation employed in the methods of the present
invention employs transdermal delivery devices ("patches"). Such transdermal
patches may be used to provide continuous or discontinuous infusion of the
compounds of the present invention in controlled amounts. The construction
and use of transdermaI patches for the delivery of pharmaceutical agents is
well
known in the art. See, e. ~. , U. S. Patent 5 ,023,252, issued June I 1, 1991,
herein incorporated by reference. Such patches may be constructed for
continuous, pulsatile, or on demand delivery of pharmaceutical agents.
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Frequently, it will be desirable or necessary to introduce the pharmaceutical
composition to the brain, either directly or indirectly. Direct techniques
usually invotve placement of a drug delivery catheter into the host's
ventricular
system to bypass the blood-brain barrier. One such implantable delivery system
used for the transport of biological factors to specific anatomical regions of
the
body is described in U. S . Patent 5 , O 11,472 which is herein incorporated
by
reference.
Indirect techniques, which are generally preferred, usually involve
formulating the compositions to provide for drug latentiation by the
conversion
of hydrophilic drugs into lipid-soluble drugs. Latentiation is generally
achieved
through blocking of the hydroxy, carbonyl, sulfate, and primary amine groups
present on the drug to render the drug more lipid soluble and amenable to
transportation across the blood-brain barrier. Alternatively, the delivery of
hydrophilic drugs may be enhanced by intra-arterial infusion of hypertonic
solutions which can transiently open the blood-brain barrier.
Other suitable formulations for use in the present invention can be found in
Remington 's Pharmaceutical Sciences, Mace Publishing Company, Philadelphia,
PA, 17th ed. (1985).
Utility
The compounds and pharmaceutical compositions of the invention are
useful in inhibiting (3-amyloid peptide release and/or its synthesis, and,
accordingly, have utility in diagnosing and treating Alzheimer's disease in
mammals including humans.
As noted above, the compounds described herein are suitable for use in a
variety of drug delivery systems described above. Additionally, in order to
enhance the in vivo serum half-life of the administered compound, the
compounds may be encapsulated, introduced into the lumen of liposomes,
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prepared as a colloid, or other conventional techniques may be employed which
provide an extended serum half life of the compounds. A variety of methods
are available for preparing liposomes, as described in, e.g., Szoka, et al.,
U.S.
Patent Nos. 4,235,871, 4,501,728 and 4,837,028 each of which is incorporated
herein by reference.
The amount of compound administered to the patient will vary depending
upon what is being administered, the purpose of the administration, such as
prophylaxis or therapy, the state of the patient, the manner of
administration,
and the like. In therapeutic applications, compositions are administered to a
patient already suffering from AD in an amount sufficient to at least
partially
arrest further onset of the symptoms of the disease and its complications. An
amount adequate to accomplish this is defined as "therapeutically effective
dose. " Amounts effective for this use will depend on the judgment of the
attending clinician depending upon factors such as the degree or severity of
AD
in the patient, the age, weight and general condition of the patient, and the
like.
Preferably, for use as therapeutics, the compounds described herein are
administered at dosages ranging from about 1 to about 500 mg/kg/day.
In prophylactic applications, compositions are administered to a patient at
risk of developing AD (determined for example by genetic screening or familial
trait) in an amount sufficient to inhibit the onset of symptoms of the
disease.
An amount adequate to accomplish this is defined as "prophylactically
effective
dose. " Amounts effective for this use will depend on the judgment of the
attending clinician depending upon factors such as the age, weight and general
condition of the patient, and the like. Preferably, for use as prophylactics,
the
compounds described herein are administered at dosages ranging from about 1
to about 500 mg/kglday.
As noted above, the compounds administered to a patient are in the form of
pharmaceutical compositions described above. These compositions may be
...... , . ..._.,.-.. .. ~ .......
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sterilized by conventional sterilization techniques, or may be sterile-
filtered.
The resulting aqueous solutions may be packaged for use as is, or lyophilized,
- the lyophilised preparation being combined with a sterile aqueous carrier
prior
to administration. The pH of the compound preparations typically will be
between 3 and 11, more preferably from 5 to 9 and most preferably from 7 and
8. It will be understood that use of certain of the foregoing excipients,
carriers,
or stabilizers will result in the formation of pharmaceutical salts.
The compounds described herein are also suitable for use in the
administration of the compounds to a cell for diagnostic and drug discovery
purposes. Specifically, the compounds may be used in the diagnosis of cells
releasing andlor synthesizing (3-amyloid peptide. In addition the compounds
described herein are useful for the measurement and evaluation of the activity
of other candidate drugs on the inhibition of the cellular release and/or
synthesis of ,Q-amyloid peptide.
The following synthetic and biological examples are offered to illustrate
this invention and are not to be construed in any way as limiting the scope of
this invention.
EXAMPLES
In the examples below, the following abbreviations have the following
meanings. If an abbreviation is not defined, it has its generally accepted
meaning.
BEMP - 2-tert-butylimino-2-diethylamino-1,3-
dimethylperhydro-1, 3 ,2-diazaphosphorine
Boc - t-butoxycarbonyl
BOP - benzotriazol-1-yloxy-
tris(dimethylamino)phosphonium
hexafluorophosphate
bd - broad doublet
bs - broad singlet
d - doublet
dd - doublet of doublets
DIC - diisopropylcarbodiimide
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DCM - dichloromethane -
DMF - dimethylformamide
DMAP - dimethylaminopyridine
DMSO~ - - dimethylsulfoxide
EDC - ethyl-i-(3-dimethyaminopropyl)carbod_
iimide
eq. - equivalents
EtOAc - ethyl acetate
EtOH - ethanol
g - grams
HOBT - 1-hydroxybenzotriazole hydrate
Hunig's base diisopropylethylamine
-
L - liter
m - multiplet
M - molar
max - maximum
meq - milliequivalent
mg - milligram
mL - milliliter
mm - millimeter
mmol - millimole
MOC - methoxyoxycarbonyl
N - normal
NIA - not available
ng - nanogram
nm - nanometers
OD - optical density
PEPC - 1-(3-{1-pyrrolidinyl)propyl)-3-ethylcarbodiimide
PP-HOBT - piperidine-piperidine-1-hydroxybenzotrizoie
psi - pounds per square inch
- phenyl
q - quartet
quint. - quintet
rpm - rotations per minute
s - singlet
t - triplet
TFA - trifluoroacetic acid
THF - tetrahydrofuran
tlc - thin layer chromatography
~.L - microliter
UV - ultra-violet
Additionally, the following abbreviations are used to indicate the
commercial source for certain compounds and reagents:
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Aldrich - Aldrich Chemical Company, Inc. , -1001
West
- Saint Paul Avenue, Milwaukee, WI 53233
USA
Fluka - Fluka Chemical Corp. , 980 South 2nd
Street,
Ronkonkoma NY 11779 USA
Lancaster - Lancaster Synthesis, Inc., P.O. Box
100
Windham, NH 03087 USA
Sigma - Sigma, P.O. Box 14508, St. Louis MO
63178
USA
Chemservice - Chemservice Inc. , Westchester, PA
Bachem - Bachem Biosciences Inc. , 3700 Horizon
Drive, Renaissance at Gulph Mills, King
of
Prussia) PA 19406 USA
Maybridge - Maybridge Chemical Co. Trevillett, Tintagel,
Cornwall PL34 OHW United Kingdom
TCI - TCI America, 9211 North Harborgate Street,
Portland OR 97203
Alfa - Johnson Matthey Catalog Company, Inc.
30
Bond Street, Ward Hill, MA OI835-0747
Novabiochem - Calbiochem-Novabiochem Corp. 10933 North
Torrey Pines Road, P.O. Box 12087, La
Jolla
CA 92039-2087
Oakwood - Oakwood, Columbia, South Carolina
Advanced Chemtech
= Advanced Chemtech,
Louisville, KY
Pfaltz & Bauer - Pfaltz & Bauer, Waterbury, CT, USA
In the examples
below, all temperatures
are in degrees
Celsius (unless
otherwise indicated).Each of the compounds set forth in Examples
1- was
prepared by one
of the following
General Procedures,
unless otherwise
indicated.
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GENERAL PROCEDURE A
EDC Coupling -- Procedure I
A carboxylic acid ( 1.0 eq. ), 1-hydroxybenzotriazole hydrate ( 1.1 eq. ) and
an amine { 1. 0 eq . } were stirred in THF under a nitrogen atmosphere. To
this
mixture was added 1-(3-dimethyIaminopropyl)-3-ethylcarbodiimide
hydrochloride (1.1 eq.) followed by a suitable base, such as Hunig's base (1.1
eq. of base if a free amine is used, and 2.1 eq. of base if an amine
hydrochloride is used). After stirring from about 4 h to 17 h at room
temperature, the solvent was removed under reduced pressure. The residue was
partitioned between ethyl acetate and water and the organic layer was
separated
and washed sequentially with aqueous sodium bicarbonate, dilute aqueous
hydrochloric acid and brine. The organic layer was then dried (sodium sulfate)
and concentrated under reduced pressure. The product was either used without
further purification or was purified using standard procedures, such as silica
gel chromatography andlor recrystallization.
GENERAL PROCEDURE B
EDC Coupling -- Procedure II
A carboxylic acid (1 eq.) was stirred in a suitable solvent (such as THF,
dioxane or DMF) and an alcohol or oxime (1 - S eq.) was added. To this
mixture was added 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide
hydrochloride ( 1.2 eq. ) and 1-hydroxybenzotriazole hydrate ( 1 eq. )
followed by
a suitable base, such as 4-methylmorpholine, triethylamine, or Hunig's base
(0 - 1 eq.). A catalytic amount (0.1 eq.) of 4-dimethylaminopyridine was then
added and the mixture was stirred at ambient temperature and under a dry
atmosphere of nitrogen. After 20 hours, the mixture was concentrated under
reduced pressure and the resulting concentrate was partitioned between ethyl
acetate and water. The organic portion was separated and washed with aqueous
sodium bicarbonate and brine, and then dried (sodium sulfate) and concentrated
under reduced pressure. The crude product was either used without further
....-.... r.
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purification or was purified using standard procedures, such as silica gel
chromatography and/or recrystallization.
GENERAL PROCEDURE C
EDC Coupling -- Procedure III
To a solution of the amine (1 eq.) in THF (0.08-0.06 M) at 0°C
under an
atmosphere of nitrogen was added 3,5-difluorophenylacetic acid (1.10 eq.),
1-hydroxybenzotriazole hydrate (1.15-1.20 eq.), N,N diisopropylethyiamine
(2.30 eq.), followed by 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide
hydrochloride (1.15-1.20 eq.). The cooling bath was removed and the mixture
allowed to warm to ambient temperature with stirring for 12-14 hours. The
solution was then diluted with ethyl acetate, washed with 0.2-0.5 M aqueous
HCl (2x), dilute aqueous NaHC03 (lx), and brine (lx), and then the organic
phase was dried over Na~S04, filtered, concentrated in vacuo, and the residue
purified by flash chromatography to afford the product.
GENERAL PROCEDURE D
Removal of N ten-BOC Protecting Group -- Procedure I
To the N Boc-protected imidazoline (1 eq.) in a round-bottomed flask,
cooled to 0°C, was added trifluoroacetic acid (0.07-0.08 M) followed by
anisole (6.2-8.0 eq.). The resulting solution was stirred at 0°C for 1
hour, then
allowed to warm to ambient temperature with stirring for 1-1.5 hours. The
solution was then concentrated in vacuo, Et20 was added and the mixture again
concentrated in vacuo. The residue was dissolved in either CH,C12 or EtOAc
and washed with dilute aqueous NaHCO~ ( 1 or 2x) , and sometimes brine ( 1 x),
then the organic phase was dried over Na2S04, filtered, and concentrated in
vacuo to afford the product.
GENERAL PROCEDURE E
Removal of N tent-BOC Protecting Group -- Procedure II
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The N ten-Boc-amine was dissolved in a suitable dry solvent (such as 1,4-
dioxane or ethyl acetate) and the solution was cooled in an ice bath. Gaseous
HCl was introduced into the solution until the mixture was saturated with HCl
and the mixture was then stirred until the starting material was consumed. The
resulting mixture was concentrated under reduced pressure to yield the amine
hydrochloride. The amine hydrochloride was either used without further
purification or was triturated in diethyl ether and the resulting solid
collected by
filtration.
GENERAL PROCEDURE F
Thiazoline Synthesis -- Procedure I
To a stirred solution of a nitrite (1 eq.) in dry ethanol under an atmosphere
of nitrogen was added 2-mercaptoethylamine ( 1 - 3 eq. ) . The reaction
mixture
was then heated at reflux until the nitrite was consumed. The mixture was then
concentrated under reduced pressure and the resulting product purified by
chromatography andlor recrystallization.
GENERAL PROCEDURE G
Thiazoline Synthesis -- Procedure II
To a stirred solution of a nitrite ( 1 eq. ) in dry ethanol under an
atmosphere
of nitrogen were added L-cysteine ethyl ester hydrochloride (1 - 3 eq.)
(Aldrich) and a stoichiometric amount of N,N diisopropylethylamine (based on
the L-cysteine ethyl ester hydrochloride) . The reaction mixture was heated at
reflux for three hours and then concentrated under reduced pressure. The
resulting product was purified by chromatography and/or crystallization.
GENERAL PROCEDURE H
Cyclization Using Burgess Reagent
To a solution of N [N (3,5-dichlorophenyl)-L-alaninyl]-L-serine methyl
ester (178 mg, 0.5 mmol) in 2.5 mL of THF was added 147 mg (0.62 mmol,
1.2 eq. ) of (methoxycarbonylsulfamoyl)triethylammonium hydroxide, inner salt
r.
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(available from Aldrich Chemical Co., Milwaukee, Wisconsin). The solution
was refluxed for 5 h and then stirred at room temperature for 36 h. The
solvents were then removed and the residue purified by preparative tlc using
3:2 hexanes/ethyl acetate as the eluent to afford 50 mg of 2-[(S)-1-(3,5-
' 5 dichloroanilino)ethyl]-(S)-4-methoxycarbonyl-2-oxazolidine as a yellow
oil.
Various other (S)-4-alkoxycarbonyl-2-oxazolidines can be prepared from N-
substituted (amino acid)-L-serine esters using this procedure.
GENERAL PROCEDURE I
S-AminoaIkvl-1,2,4-Oxadiazole Synthesis
An N tent-Boc-protected amino acid and an amidoxime are reacted
according to General Procedure B above to provide the corresponding O-
acyloxime. The O-acyloxime is then heated at reflux in xylenes with the
azeotropic removal of water to provide the N ten-Boc-protected 5-aminoalkyl-
1,2,4-oxadiazole derivative, which is then typically purified by silica gel
chromatography. The tert-Boc protecting group is then removed according to
General Procedure E above to afford the 5-aminoalkyl-1,2,4-oxadiazole.
The following Examples A-L illustrate the synthesis of heterocyclic
intermediates which may be used to prepare the compounds of the present
invention.
Example A
Synthesis of
(S)-5-(1-Aminoethyl)-3-ethyl-1,2,4-oxadiazole Hydrochloride
Following General Procedure I above, the title compound was prepared
using N tent-Boc-L-alanine (Sigma) and propanamidoxime.
Example B
Synthesis of
(S)-5-(1-Amino-2-phenylethyl)-3-methyl-1,2,4-oxadiazole Hydrochloride
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Following General Procedure I above, the title compound was prepared
using N tent-Boc-L-phenylalanine (Sigma) and acetamidoxime.
Example C
Synthesis of
(S)-5-(1-Amino-1-phenylmethyl)-3-methyl-1,2,4-oxadiazole Hydrochloride
Following General Procedure I above, the title compound was prepared
using N tent-Boc-L-phenylglycine (Sigma) and acetamidoxime.
Example D
Synthesis of
(S)-5-(1-Amino-1-phenylmethyl)-3-phenyl-1,2,4-oxadiazole Hydrochloride
Following General Procedure I above, the title compound was prepared
using N tent-Boc-L-phenylglycine (Sigma) and benzamidoxime.
Example E
Synthesis of (S)-5-(1-Amino~l-phenylmethyl)
3-(4-methoxyphenylmethyl)-1,2,4-oxadiazole Hydrochloride
Following General Procedure I above, the title compound was prepared
using N tent-Boc-L-phenylglycine {Sigma) and 4-methoxyphenylacetamidoxime.
Example F
Synthesis of
(2S)-2-(1-Aminoethyl)-5(R,S)-ethoxycarbonyl-2-oxazoline
Step A -- Synthesis of N-Carbobenzyloxy-L-alanine-D,L-isoserine Ethyl
Ester
Following General Procedure A above and using N carbobenzyloxy-L
alanine (Sigma) and D,L-isoserine ethyl ester hydrochloride, the title
compound
was prepared.
Step B -- Synthesis of (2S)-2-[1-(N Carbobenzyloxy)aminoethyl]-5(R,S)-
ethoxycarbonyl-2-oxazoline
_... _. ~_~....~.... ..._ ... ..
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To a solution of N carbobenzyloxy-L-alanine-D,L-isoserine ethyl ester from
Step A above in dry THF was added triphenylphosphine (2 eq. ) . The mixture
was cooled'fo 0°C and a solution of diethyl azodicarboxylate (2 eq.) in
THF
was added dropwise. The resulting mixture was allowed to warm to room
S temperature and, after 20 hours, the mixture was concentrated under reduced
pressure. The resulting oil was purified by silica gel chromatography to yield
the title compound as an oil which was used without further purification.
Step C -- Synthesis of (2S)-2-(1-Aminoethyl)-5(R,S}-ethoxycarbonyl-2-
oxazoline
The crude oil from Step B above was dissolved in ethanol and the solution
was degassed and stirred under an atmosphere of nitrogen. Palladium on
carbon ( 10 % ) was added and the atmosphere of nitrogen was replaced with
hydrogen under balloon pressure. The mixture was stirred for 2 hours and then
filtered through Celite. The filtrate was concentrated and the residue
purified
by silica gel chromatography to provide the title compound as an oil.
Example G
Synthesis of
(S)-2-(1-Aminoethyl)-4(R,S)-ethoxycarbonyl-2-thiazoline Hydrochloride
Step A -- Synthesis of (S)-2-[1-(N-tent-Butoxycarbonyl)aminoethyl]-4-
ethoxycarbonyl-2-thiazoline
The title compound was prepared according to the procedures described in
C. D. J. Boden, et al., Synlett, 5, 417 (1995).
Step B -- Synthesis of (S)-2-(1-Aminoethyl)-4(R,S}-ethoxycarbonyl-2-
thiazoline Hydrochloride
The N tent-Boc protecting group was removed from the product of Step A
above using General Procedure E above to afford the title compound.
Example H
Synthesis of
2-(3,5-Difluorophenylmethyl)-4-carboxy-2-thiazoline
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Step A -- Synthesis of 2-(3, 5-Difluorophenylmethyl)-5-ethoxycarbonyl-2-
thiazoline
Following General Procedure G and using 3,5-difluorophenylacetonitrile
and L-cysteine ethyl ester hydrochloride (Aldrich), the title compound was
prepared.
Step B -- Synthesis of 2-(3,5-Difluorophenylmethyl)-5-carboxy-2-
thiazoline
The title compound was prepared by hydrolysis of the product from Step A
above using 1 equivalent of LiOH in wet dioxane as described in General
Procedure II-A, Method B.
Example I
Synthesis of
N (3,5-Difluorophenylacetyl)phenylglycinonitrile
Following General Procedure A above and using 3,5-difluorophenylacetic
acid {Aldrich) and 2-phenylglycinonitrile hydrochloride (Lancaster), the title
compound was prepared. The crude product was purified by silica gel
chromatography using 1:1 ethyl acetatelhexanes as the eluent, followed by
crystallization from 1-chlorobutane/acetonitrile.
NMR data was as follows:
1H-nmr {CDC13, 250 MHz): 8 = 3.61 (s, 2H), 6.13 (d, 1 H), 6.98 (m, 2H),
7. I 2 (m, 1 H), 7.46 (m, SH), 9.40 (d, 1 H).
C,6H~~FZN,O (MW = 286.28); mass spectroscopy (M+) 286.
Example J
Synthesis of 2-(1-Aminoethyl)-1-tert-butoxycarbonyl
4-methoxycarbonyl-4-phenylmethyl-2-imidazoline
Step A -- Synthesis of 1-[2-(N carbobenzyloxy)amino-
thiopropionyl]piperidine
Following the procedures described in Gilbert, et al., Tetrahedron, 1995 ~ I ,
6315-6336 and using N carbobenzyloxy-D,L-alanine {Sigma), the title compound
was prepared in two steps (51 % overall yield) as a solid having a melting
point
_..___r.4... . r
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of 68-69°C. The final reaction was monitored- by tlc (Rf = 0.59 in 24:1
DCM/EtOAc) and the product was purified by flash column chromatography.
NMR data was as follows:
'H-nmr (CDCl3, 250 MHz): b = 7.37-7.29 (m, SH), 6.46 (lrd, 1H, J= 8.26
Hz), 5. I 0 (ABq, 2H, JAB= 12.38 Hz, avAB= 9.12 Hz), 4.92 (p, 1 H, J= 7.07
Hz),
4.41-4.34 (m, 1H), 4.17-4.06 (m, 1H), 3.86-3.65 (m, 2H), 1.75-1.65 (bm, 6H),
1.35 (d, 3H, J= 6.75 Hz).
C,6H"NzO~S (MW = 306.43); mass spectroscopy (MH') 306.2.
Step B -- Synthesis of Methyl 2,3-Diamino-2-phenylmethylpropionate
Following the procedures described in Gilbert, et al., Tetrahedron, 1995 51,
6315-6336 and using L-phenylalanine methyl ester hydrochloride (Aldrich), the
title compound was prepared in three steps ( 14% overall yield). The final
reaction was monitored by tlc (Rf = 0.26 in 9:1 DCM/MeOH) and the product
was purified by flash column chromatography, followed by recrystallization
from cyclohexane/ethyl acetate (20: I ).
NMR data was as follows:
'H-nmr (CDCl3, 250 MHz): 8 = 7.32-7.17 (m, 3H}, 7.14-7.09 (m, 2H), 3.70
(s, 3H), 2.96 (ABq, 2H, JAB = 13.13 Hz, OvAB= 114.63 Hz), 2.90 (ABq, 2H, JAB=
13.38 Hz, wAB= 86.35 Hz), 1.55 (bs, 4H).
C"H,6N~02 (MW = 208.26); mass spectroscopy (MH+) 209.1.
Step C -- Synthesis of 2-[1-(N Carbobenzyloxy)aminoethyl)-4-
methoxycarbonyl-4-phenylmethyl-2-imidazoline
The product from Step A above (1 equivalent) in iodomethane (35
equivalents) was heated to reflux under nitrogen for 8 hours. The dark yellow
solution was concentrated in vacuo to a golden yellow foam, then azeotroped
with methanol (2x) to remove any residual iodomethane. The residue was
dissolved in methanol (0.2 M) and the product from Step B above ( 1
equivalent)
was added. The resulting clear colorless solution was heated to reflux for 1
hour under nitrogen and then stirred at ambient temperature for 14 hours, and
again heated to reflux for an additional 1 hour. The solution was allowed to
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cool to ambient temperature and then concentrated in vacuo. The reaction was
monitored by tlc (Rf = 0.27 in 95:5 DCM/MeOH) and the residue was purified
by flash col~rmn chromatography to provide the title compound as a 1:1 mixture
of diastereomers (75% yield). -
NMR data was as follows:
'H-nmr {CDC13, 250 MHz): 8 = 7.36-7.20 (m) 8H), 7.14-7.08 (m, 2H),
5.84-5.74 {m, 1 H), 5.15-5.02 (m, 2H), 4.96 (bs, 1 H}, 4.44-4.30 (m, 1 H),
4.03-
3.97 (m, 1H), 3.72-3.66 (m, 1H}, 3.70 (s, 1.5H), 3.69 {s, 1.SH), 3.07 (ABA,
1H,
JAB= 13.38 Hz, OvAB= 54.25 Hz), 3.06 (ABq, 1H, JAB= 13.51 Hz, wAB= 52.03
Hz), 1.39 (d, 1.5H, J= 7.00 Hz), 1.35 (d, 1.5H, J= 7.00 Hz).
C"H,5NjO4 (MW = 395.46); mass spectroscopy (MH~) 396.1.
Step D -- Synthesis of 1-tert-Butoxycarbonyl-2-[1-(N
carbobenzyloxy)aminoethyl]-4-methoxycarbonyl-4-phenylmethyl-2-
imidazoline
To a 1:1 mixture of the product from Step C above (1.815 g, 1 eq.) in THF
(9 mL)/H20 (9 mL) at ambient temperature was added NaHC03 (0.377 g, 1.50
eq.) and di-tert-butyl dicarbonate (I.304 g, 2.00 eq.) (Aldrich) in THF (6
mL).
The resulting pale yellow mixture was stirred at ambient temperature for 1
hour
and then additional NaHC03 (0.188 g, 0.75 eq.) and di-tert-butyl dicarbonate
(0.652 g, 1.00 eq.) in THF (3 mL) were added and the mixture was stirred for
an additional hour. The mixture was then diluted with ethyl acetate, washed
with brine (2x), dried over NaZS04, filtered, concentrated in vacuo, and
purified
by flash column chromatography using 3:2 hexanes/EtOAc as the eluent to
provide the title compound in 77% yield as a viscous oil (Rf = 0.31 in 3:2
hexanes/EtOAc). The racemic product was isolated as a 1:1 mixture of
diastereomers.
NMR data was as follows:
'H-nmr (DMSO-d3, 250 MHz): c~ = 7.38-7.09 (m, lOH), 6.00 (bd, 1H, J=
7.75 Hz}, 5.21-5.05 (m, 3H), 4.15 (d, 1 H, J= 11.76 Hz), 4.08 (d, 1 H, J=
11.26
Hz), 3.85-3.80 (m, 2H), 3.78 (s, 3H), 3.20-3.00 (m, 2H), 1.43 (s, 9H), 1.36
(d,
1.5H, J= 6.75 Hz), 1.24 (d, 1.5H, J= 6.75 Hz).
..~...~ ..~_ _... ,
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-- 1 I7 --
C,,H33N3O6 (MW = 495.58); mass spectroscopy (MH'~} 496.4. -
Step E =- Synthesis of 1-tert-Butoxycarbonyl-2-(1-aminoethyl)-4-
methoxycarbonyl-4-phenylmethyl-2-imidazoline
A mixture of the racemic N Cbz-protected amine from Step D above and
20% Pd{OH)z on carbon (20% weight eq.) in MeOH (0.010-0.015 M) under an
atmosphere of hydrogen (35-40 psi) were shaken for 4 hours. The catalyst was
removed by filtration through a plug of Celite and the filtrate was
concentrated
in vacuo to provide the title compound as a viscous oil (Rf = 0.26 in 95:5
DCM/MeOH).
NMR data was as follows:
'H-nmr (CDCI~, 250 MHz): 8 = 7.32-7.15 (m, SH), 4.19-4.05 (m, 2 H),
3.87-3.78 (m, 1 H), 3.80 (s, 3H), 3.21-3.03 (m, 2H), I .98 (bs, 2H), 1.43 (s,
9H),
1.33 (d, 1.SH, J= 7.00 Hz), 1.26 (d, 1.SH, J= 6.50 Hz).
IS C~9H,~N3O4 (MW = 361.44); mass spectroscopy (Ml-1+) 361.
Example K
Synthesis of 2-(1-Aminoethyl)-1-tert-butoxycarbonyl
4-methoxycarbonyl-4-phenyl-2-imidazoline
Step A -- Synthesis of Methyl 2,3-Diamino-2-phenylpropionate
Following the procedures described in Gilbert, et al., Tetrahedron, 1995 51,
6315-6336 and using (S)-(+)-2-phenylglycine methyl ester hydrochloride
(Aldrich), the title compound was prepared in three steps ( 16% overall
yield).
The final reaction was monitored by tlc (Rf = 0.34 in 9:1 CHzCI,/MeOH) and
the product was purified by flash column chromatography.
NMR data was as follows:
'H-nmr (CDC13, 250 MHz): 8 = 7.52-7.47 (m, 2H), 7.39-7.25 (m, 3H), 3.74
(s, 3H), 3.42 (d, 1 H, J = 13.26 Hz), 2.89 {d, 1 H, J = 13.26 Hz), 1.88 (bs,
4H).
C,oH,4Nz0z (MW = 194.23); mass spectroscopy (MH') 195.
Step B -- Synthesis of 2-[1-(N Carbobenzyloxy)aminoethyl)-4-
methoxycarbonyl-4-phenyl-2-imidazoline
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The product from Example J -- Step A (5:206 g, 1.1 eq.) in iodomethane
(36.5 mL, 38 eq.) was heated to reflux under nitrogen for 16 hours. The dark
yellow solution was concentrated in vacuo to a golden yellow foam, then
azeotroped once with methanol (30 mL) to remove any residual iodomethane.
The residue was dissolved in methanol (30 mL) and the product from Step A
above (3.00 g, 1 eq.) was added. The resulting pale yellow solution was heated
to reflux under nitrogen for 3.5 hours. The solution was then allowed to cool
to
ambient temperature, concentrated in vacuo, and purified by flash
chromatography to yield (5.08 g, 86%) of the title compound as a
1:1 mixture of racemic diastereomers. The reaction was monitored by tlc (Rf =
0.32 in 95:5 CH,CI,/MeOH) and the product was purified by flash column
chromatography.
NMR data was as follows:
'H-nmr (CDCl3, 250 MHz): 8 = 7.38-7.28 (m, lOH), 6.09-6.00 (m, 1H),
5. I 1-5.04 (m, 2H), 4.10 (bs, I H), 3.84-3.68 (m, 4H), 1.53 (d, 3H, J = 7.00
Hz).
C.,oH,3N3O4 (MW = 381.43); mass spectroscopy (MH+) 382.4
Step C -- Synthesis of 1-tert-Butoxycarbonyl-2-(1-(N
carbobenzyloxy)aminoethylJ-4-methoxycarbonyl-4-phenyl-2-imidazoline
To product from Step B above (4.94 g, 1 eq.) in THF (35 mL)/H20 (35
mL) at ambient temperature was added NaHC03 (2.00 g, 1.84 eq.) and di-tert-
butyl dicarbonate (7.50 g, 2.65 eq.) (Aldrich). The resultant pale yellow
mixture
was stirred at ambient temperature for 1 hour and then additional NaHC03 (0.45
g, 0.41 eq.) and di-tert-butyl dicarbonate (0.98 g, 2.65 eq.) were added and
the
mixture was stirred for an additional hour. The mixture was then diluted with
ethyl acetate, washed with H,O ( 1 x), brine (2x), dried over Na~S04,
filtered,
concentrated in vacuo, and purified by flash chromatography. The racemic
diastereomeric products [Rf 0.38 (isomer A as a racemic mixture) and 0.28
(isomer B as a racemic mixture) eluting with 6:1 toluene/EtOAc] were separated
by flash chromatography using 6:1 toluene/EtOAc with a one step gradient to
5:1 toluene/EtOAc as the eluent to provide two sets of racemic enantiomer
pairs
.....~ ._ ._. _ ~
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in 73% yield, isomer A ( 1.122 g) as a viscous oil and isomer B ( 1.0868 g) as
a
white solid.
' NMR data was as follows (isomer A):
'H-nmr (CDCl3, 250 MHz): S = 7.39-7.27 (m, lOH), 6.20 (bd, 1H, J= 8.00
S Hz), 5.40-5.30 (m, 1 H), 5.13 (s,. 2H), 4.84 (d, 1 H, J= 1 I .26 Hz), 3.87
(d, 1 H, J=
11.26 Hz), 3.70 (s, 3H), 1.52-1.50 (m, 12H).
C,6H3,N3O6 (MW = 481.55); mass spectroscopy (MH+) 482.3.
Step D -- Synthesis of 1-tert-Butoxycarbonyl-2-(1-aminoethyl)-4-
methoxycarbonyl-4-phenyl-2-imidazoline
Isomer A of Step C above and 20% Pd(OH), on carbon (20% weight eq.) in
methanol (0.010-0.015 M) under an atmosphere of hydrogen (35-40 psi) were
shaken for 4 hours. The catalyst was removed by filtration through a plug of
Celite and the filtrate was concentrated in vacuo and purified by flash
chromatography to provide the title compound (88%) as a viscous oil (Rf = 0.28
in 95:5 DCM/MeOH).
NMR data was as follows:
'H-nmr (CDC13, 250 MHz): 8 = 7.43-7.27 (m, SH), 4.80 (d, 1H, J= 11.26
Hz), 4.41 (q, 1 H, J= 6.75 Hz), 3.87 (d, 1 H, J= 11.51 Hz), 3.73 (s, 3H), 2.02
(s,
2H), 1.49 (s, 9H), 1.45 (d, 3H, J= 6.75 Hz).
C~BHZSN3O4 (MW = 347.42); mass spectroscopy (MH+) 348.2.
Example L
Synthesis of
(4R)-4-Carboxy-2-(3,5-difluorophenylmethyl)-4-methyl-2-thiazoline
Step A -- Synthesis of Methyl 3,5-Difluorophenylacetimidate
Hydrochloride
. The title compound was prepared according to the procedures described in
S. C. Zimmerman, et al., J Org. Chem. 1989, 54, 1256-1264, for the synthesis
of imidates. Specifically, a stream of anhydrous HCl gas was passed through a
solution of 3,5-difluorophenylacetonitrile (5.045 g, 1 eq.) (Lancaster) and
methanol (2.00 mL, 1.5 eq.) in diethyl ether (100 mL) at 0 °C for 20
minutes.
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The solution was then allowed to warm to ambient temperature under nitrogen
with stirring for 23 hours, during which time a white precipitate formed. The
white precipitate was collected by filtration and rinsed with diethyl ether to
yield
the title compound (6.6756 g, 91 %) as a solid having a melting point of I
56.5-
157.5°C.
NMR data was as follows:
'H-nmr (CDC13, 300 MHz): 8 = 13.01 (bs, i H), 11.93 {bs, 1 H), 7.04-6.97
(m, 2H), 6.83-6.75 (m, 1H), 4.30 (s, 3H), 4.08 (s, 2H).
Step B -- Synthesis of Methyl (R)-2-Methylcysteine Hydrochloride
The title compound was prepared as a viscous oil in five steps (35% overall
yield) from D-(S)-cysteine methyl ester hydrochloride as described in G.
Pattenden, et al., Tetrahedron, 1993, 49, 2131-2138 and references cited
therein.
D-(S)-cysteine methyl ester hydrochloride was prepared in one step (98% yield)
from D-cysteine hydrochloride monohydrate (Aldrich) as described in J. E.
Baldwin, et al., Tetrahedron, 1989, 45, 4537-4550.
NMR data was as follows:
'H-nmr (D,O, 250 MHz): 8 = 4.83 (bs, 4H), 3.92 (s, 3H), 3.27 (d, 1H, J=
15.26 Hz), 3.02 (d, 1H, J= 15.01 Hz), 1.69 (s, 3H).
Optical Rotation: [a]~o = 2.44 (c 1.15, 1-1~0)
CSH,,NO,SCI (MW = 185.67); mass spectroscopy (MH+) 149.1.
Step C -- Synthesis of (4R)-2-(3,5-Difluorophenylmethyl)-4-
methoxycarbonyl-4-methyl-2-thiazoline
The title compound was prepared according to the procedures described in
G. Pattenden, et al., Tetrahedron, 1995, 51, 7313-7320, for the synthesis of
thiazolines. Specifically, to a solution of methyl (R)-2-methylcysteine
hydrochloride (1.7109 g, 1.00 eq.) from Step B above in CHZCI, (55 mL) at
ambient temperature under nitrogen was added the imidate hydrochloride (2.042
g, 1.00 eq.) from Step A above followed by the dropwise addition of
triethylamine (1.28 mL, 1.00 eq.) over a period of 20 minutes. The resulting
opaque pale yellow solution was stirred at ambient temperature for 40 hours,
-__.w_"..r,.~._...__,._. .... ~
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during which time an off white suspension formed. The mixture was diluted
with CH,CI,, washed with 1 N aqueous HCI ( 1 x), H,O ( 1 x), and brine ( 1 x);
then
the organic phase was dried over MgS04, filtered, concentrated in vacuo, and
the residue purified by flash chromatography using 20:1 DCM/EtOAc as the
eluent to afford the title compound ( 1.6669 g, 63%) as a colorless oil (Rf =
0.34
in 20:1 DCM/EtOAc).
NMR data was as follows:
'H-nmr (CDC13, 300 MHz): 8 = 6.85-6.79 (m, 2H), 6.74-6.67 (m, 1H), 3.81
(s, 2H}, 3.80 (s, 3H), 3.79 (d, 1 H, J= 11.30 Hz), 3.17 (d, 1 H, J= 11.30 Hz),
1.56
(s, 3H).
Optical Rotation: [a~~~, = 3.71 (c 1.02, CHCI;).
C,3H,3NO~SF, (MW = 285.32); mass spectroscopy (MH+) 285.2.
Step D -- Synthesis of (4R)-4-Carboxy-2-(3,5-dit7uorophenylmethyl)-4-
methyl-2-thiazoline
To the product from Step C above (0.7265 g, 1.0 eq.) in 1,4-dioxane (15
ml.) at ambient temperature was added LiOH ~ (0.0671 g, 1. I eq. ) in water (
1
mL) dropwise over a period of 2 minutes. Additional water (2 mL) was added
and the resulting opaque colorless solution was stirred at ambient temperature
for 1.25 hours, during which time the solution became clear and colorless. The
solution was concentrated in vacuo to a volume of approximately 5 mL and the
concentrate was diluted with ethyl acetate and washed with 1 N aqueous HC1
(2x), and water ( 1 x); then dried over Na~S04, filtered, and concentrated in
vacuo
to a give the title compound (0.6111 g, 88% yield) as a white solid having a
melting point of 119.5-122.0°C.
NMR data was as follows:
'H-nmr (CDCI,, 300 MHz): b = 10.88 (bs, 1H), 6.86-6.78 (m, 2H), 6.76-
6.69 (m, 1 H), 3.89 (ABq, 2H, JAS = 1 x.14 Hz, OvAB= 20.57 Hz), 3.84 (d, 1 H,
J=
11.48 Hz), 3.21 (d, 1 H, J= 11.61 Hz), 1.61 (s, 3H}.
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The following General Procedures II-A to II-E and Examples II=A to II-S
illustrate the synthesis of carboxylic acid intermediates which may be used to
prepare the compounds of the present invention.
GENERAL PROCEDURE II-A
Ester Hydrolysis to Free Acid
Ester hydrolysis to the free acid was conducted by conventional methods.
Below are two examples of such conventional de-esterification methods.
Method A: To a carboxylic ester compound in a 1:1 mixture of
CH30H/H,O was added 2-5 equivalents of K~CO;. The mixture was heated to
50°C for 0.5 to 1.5 hours until tlc showed complete reaction. The
reaction was
cooled to room temperature and the methanol was removed on a rotary
evaporator. The pH of the remaining aqueous solution was adjusted to ~2, and
ethyl acetate was added to extract the product. The organic phase was then
washed with saturated aqueous NaCI and dried over MgS04. The solution was
stripped free of solvent on a rotary evaporator to yield the product.
Method B: The amino acid ester was dissolved in dioxane/water (4:1 ) to
which was added LiOH (~2 eq.) that was dissolved in water such that the total
solvent after addition was about 2:1 dioxane:water. The reaction mixture was
stirred until reaction completion and the dioxane was removed under reduced
pressure. The residue was dissolved in water and washed with ether. The layers
were separated and the aqueous layer was acidified to pH 2. The aqueous layer
was extracted with ethyl acetate. The ethyl acetate extracts were dried over
Na,S04 and the solvent was removed under reduced pressure after filtration.
The residue was purified by conventional methods (e.g., recrystallization).
GENERAL PROCEDURE II-B
Acid Chloride Preparation
~._.~ r
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3,5-Difluorophenylacetic acid (30 g, 0.174 mol) (Aldrich) was dissolved in
dichloromethane and this solution was cooled to 0°C. DMF (0.5 mL,
catalytic)
was added followed by the dropwise addition of oxalyl chloride ( I 8 mL, 0.20
mol) over a 5 minute period. The reaction was stirred for 3 h and then
rotoevaporated at reduced pressure to give an oil which was placed on a high
vacuum pump for 1 h to afford 3,5-difluorophenylacetyl chloride as a thin
yellow oil. Other acid chlorides can be prepared in a similar manner.
GENERAL PROCEDURE II-C
Schotten-Baumann Procedure
3,5-Difluorophenylacetyl chloride (from General Procedure II-B) was added
dropwise to a 0°C solution of L-alanine (Aldrich) ( 16.7 g, 0.187 mol)
in 2 N
sodium hydroxide (215 mL, 0.43 mol). The reaction was stirred for 1 h at
0°C
and then overnight at room temperature. The reaction was diluted with water
( 100 mL), then extracted with ethyl acetate (3 x I 50 mL). The organic layer
was then washed with brine (200 mL), dried over MgS04, and rotoevaporated at
reduced pressure to a residue. Recrystallization of the residue from ethyl
acetate/hexanes afforded the desired product (34.5 g, 82% yield). Other acid
chlorides may be used in this procedure to provide for intermediates useful in
this invention.
GENERAI, PROCEDURE II-D
Reductive Amination
To a solution of the arylamine in ethanol in a hydrogenation flask was
' 25 added 1 equivalent of the 2-oxocarboxylic acid ester (e. g., pyruvate
ester),
followed by 10% palladium on carbon (25 weight percent based on the
arylamine). The reaction was hydrogenated at 20 psi H, on a Parr shaker until
complete reaction was indicated by tlc (30 minutes to 16 hours). The reaction
mixture was then filtered through a pad of Celite 545 (available from Aldrich
Chemical Company, Inc.) and stripped free of solvent on a rotary evaporator.
The crude product residue was then further purified via chromatography.
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GENERAL PROCEDURE II-E -
EDC Coupling Procedure
To a 1:I mixture of the corresponding carboxylic acid and the
corresponding amino acid ester or amide in CH,CI, at O°C was added 1.5
equivalents triethylamine, followed by 2.0 equivalents hydroxybenzotriazole
monohydrate and then 1.25 equivalents of ethyl-3-(3-dimethylamino)propyl
carbodiimide~HCl. The reaction mixture was stirred overnight at room
temperature and then transferred to a separatory funnel. The mixture was
washed with water, saturated aqueous NaHCO;, 1N HCl and saturated aqueous
NaCI, and then dried over MgS04. The resulting solution was stripped free of
solvent on a rotary evaporator to yield the crude product.
Example II-A
Synthesis of N-(Phenylacetyl)-L-alanine
Using General Procedure II-C, the title compound was prepared from
phenylacetyl chloride (Aidrich) and L-alanine (Aldrich) as a solid having a
melting point of 102-104°C.
NMR data was as follows:
'H-nmr {CDC13): 8 = 9.14 (br s, 1H), 7.21-7.40 (m, 5H), 6.20 (d, J = 7.0
Hz, 1H), 4.55 (m, 1H), 3.61 (s, 2H), 1.37 (d, J = 7.1 Hz, 3H).
13C-nmr (CDC13): ~ = 176.0, I ? 1.8, 134.0, 129.4, I 27.5, 48.3, 43.2, 17.9.
Example II-B
Synthesis of N-(3,5-Difluorophenylacetyl)-L-alanine
Using General Procedure II-C, the title compound was prepared from 3,5-
difluorophenylacetyl chloride (General Procedure II-B) and L-alanine
(Aldrich).
NMR data was as follows:
'H-nmr (CD30D): 8 = 8.32 (br s, 0.3H), 6.71 (m, 2H), 6.60 (m, 1H), 4.74
(br s, I .7H), 4.16 (m, 1 H), 3.36 (s, 2H), 1.19 (d, J = 7.3 Hz, 3H).
' 3C-nmr (CD30D): 8 = 175 .9, 172.4, 164.4 (dd, J = 13.0, 245.3 Hz), 141.1,
I 13.1 (dd, J = 7.8, 17.1 Hz), 102.9 (t, J = 25.7 Hz), 49.5, 42.7, 17.5.
.._..w...~"
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Example II-C -
Synthesis of N-(Cyclopentylacetyl)-L-phenylglycine
Step A = Preparation of N-(Cyclopentylacetyl)-L-phenyl~lycine Methyl Ester
Following General Procedure II-E above using cyclopentylacetic acid
{Aldrich) and L-phenylglycine methyl ester hydrochloride (Novabiochem}, the
title compound was prepared as a solid having a melting point of 83-
86°C. The
reaction was monitored by tlc on silica gel (Rf = 0.28 in 25% ethyl
acetate/hexanes) and purification was by recrystallization from ethyl
acetatelhexanes.
NMR data was as follows:
~H-nmr (CDC13): b = 7.35 (s, SH), 6.44 (bd, 1H), 5.6 (d, 1H), 3.72 (s, 3H),
2.24 {bs, 3H), 1.9-1.4 (m, 6H), 1.2-1.05 (m, 2H).
'3C-nmr (CDCI3): 8 = 172.3, 171.7, 136.7, 129.0, 128.6, 127.3, 56.2, 52.7,
42.5, 36.9, 32.40, 32.38, 24.8.
C,~HZ,N03 (MW = 275.35}; mass spectroscopy (M+Na) 298.
Step B-Preparation of N-(Cvclohentvlacetvl)-L-phenvl~lvcine
Following General Procedure II-A above using N-(cyclopentylacetyl)-L
phenylglycine methyl ester {from Step A), the title compound was prepared as a
solid having a melting point of 155-158°C. The reaction was monitored
by tlc
on silica gel (Rf = 0.18 in 10% methanol/dichloromethane).
NMR data was as follows:
'H-nmr (CDCI3): 8 = 8.60 (d, J = 7.8 Hz, 1H), 7.45 (m, SHO, 5.41 (d, J =
7.2 Hz, 1H), 2.20 (m, 3H), 1.8-1.1 (m, 8H)..
'3C-nmr (CDC13): b = 172.3, 172.0, 137.5, 128.7, 128.1, 127.8, 56.2, 40.9,
36.8, 31.8, 24.5.
C15H19N~3 (MW = 261.32); mass spectroscopy (M+Na) 284.
Example II-D
Synthesis of N-(Cyclopentylacetyl)-L-alanine
Step A - Preparation of N-(Cyclopentylacetyll-L-alanine Meth 1 Ester
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Following General Procedure II-E above using cyclopentylacetic acid
{Aldrich) and L-alanine methyl ester hydrochloride (Sigma), the title compound
was prepared as a solid having a melting point of 43-46°C. Purification
was by
recrystallization from ethyl acetate/hexanes.
NMR data was as follows:
'H-nmr (CDC13): ~ = 6.38 (d, 1H), 4.50 (m, 1H), 3.65 (s, 3H), 2.13 (bs,
3H), 1.80-1.00 (m (includes d at 1.30, 3H), 11H).
'3C-nmr (CDCI3): 8 = 173.7, 172.5, 52.1, 47.6, 42.3, 36.8, 32.15, 32.14,
18Ø
lO C"H,9NO3 (MW = 213.28); mass spectroscopy (MH') 214.
Step B - Preparation of N-(Cyclopentylacet~)-L-alanine
Following General Procedure II-A above using N-(cyclopentylacetyl}-L-
alanine methyl ester (from Step A), the title compound was prepared. The
reaction was monitored by tlc on silica gel (Rf = 0.18 in 10%
methanol/dichloromethane).
NMR data was as follows:
'H-nmr (DMSO-db): 8 = 12.45 (bs, 1H), 8.12 (d, J=7.2 Hz, IH), 4.24
(quint, J = 7.2 Hz, 1 H), 2.14 (m, 3H), 1.8-1.4 (m, 6H), 1.29 (d, J = 7.2 Hz,
3H), 1.2-1.0 (m, 3H).
'3C-nmr (DMSO-db): 8 = 174.6, 171.9, 47.3, 41.1, 36.7, 31.8, 24.5, 17.2.
C",H,.,N03 (MW = 199.25); mass spectroscopy (MH+) NIA.
Example II-E
Synthesis of N-(Cyclopropylacetyl)-L-alanine
Step A - Preparation of N-(Cyclopropylacetvl)-L-alanine Methyl Ester
Following General Procedure II-E above using cyclopropylacetic acid
(Aldrich) and L-alanine methyl ester hydrochloride (Sigma), the title compound
was prepared as an oil. The reaction was monitored by tIc on silica gel (Rf =
0.15 in 25% ethyl acetate/hexanes) and purification was by flash column
chromatography using 25% ethyl acetate/hexanes as the eluant.
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NMR data was as follows:
'H-nmr (CDCl3): S = 6.60 (d, 1H), 4.55 (m, 1H), 3.69 (s, 3H), 2.10 (m,
- 2H), 1.34 (d; 3H), 0.95 (m, 1H), 0.58 (m, 2H), 0.15 (m, 2H).
'3C-nmr (CDC13): 8 = 173.7, 172.3, 52.3, 47.7, 41.0, 18.2, -6.7, 4.27, 4.22.
C9H,SN03 {MW = 185.22); mass spectroscopy (MH~) NIA.
Step B - Preparation of N-(Cyclopent l~tyl)-L-alanine
Following General Procedure II-A above using N-(cyclopropylacetyl)-L-
alanine methyl ester (from Step A), the title compound was prepared as an oil.
The reaction was monitored by tlc on silica gel (Rf = 0.27 in i 0%
methanol/dichloromethane).
NMR data was as follows:
'I-I-nmr (DMSO-d6): 8 = 8.18 (d, 1 H), 4.25 (m, 1 H), 2.08 (m, 2H), 1.30 (d,
3H), 1.00 (m, 1 H), 0.50 (m, 2H), 0.19 (m, 2H).
'3C-nmr (DMSO-d6): 8 = 174.6, I7I.7, 47.4, 17.3, 7.6, 4.12, 4.06.
CgH,3N03 (MW = 199.25); mass spectroscopy (MH+) NIA.
Example II-F
Synthesis of N-(Cyclopropylacetyl)-L-phenylglycine
Step A - Preparation of N-(C~progylacetyl)-L-clycine Methyl Ester
Following General Procedure II-E above using cyclopropylacetic acid
(Aldrich) and L-phenylglycine methyl ester, the title compound was prepared as
a solid having a melting point of 74-76°C. The reaction was monitored
by tlc
on silica gel (Rf = 0.61 in 50% ethyl acetate/hexanes) and purification was by
recrystallization from ethyl acetate/hexanes.
NMR data was as follows:
'H-nmr (CDC13): 8 = 7.35 (m, 5H), 6.97 (bd, J=7.2 Hz, IH), 5.59 (d, J=7.8
Hz, 1 H), 3 .71 (s, 3 H), 2.17 (m, 2H), 1.05-0.95 (m, 1 H), 0.62 (m, 2H), 0.20
(rn,
2H).
'3C-nmr (CDC13): 8 = 171.9, 174.6, 136.6, 129.0, 128.5, 127.2, 56.1, 52.7,
41.0,6.9,4.37,4.33.
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C,4H,.,N03 (MW = 247.30); mass spectroscopy (MH~) N/A.
Step B'- Preparation of N-(Cvclonentylacetvl)-L-phenvlslvcine
Following General Procedure II-A above using N-(cycloprtopylacetyl)-L-
phenylgiycine methyl ester (from Step A), the title compound was prepared as a
solid having melting point of 152-157°C. The reaction was monitored by
tlc on
silica gel (Rf = 0.23 in 10% methanol/dichioromethane) and purification was by
recrystallization from ethyl acetatelhexanes.
NMR data was as follows:
'H-nmr {CDC13): 8 = 8.47 (d, J = 7.69 Hz, 1H), 7.35 (m, 5H), 5.34 (d, J =
7.69 Hz, 1 H), 2.10 (m, 2H), 0.90 (m, 1 H), 0.40 (m, 2H), 0.10 (m, 2H).
'3C-nmr (CDC13): 8 = 172.3, 171.8, 137.6, 128.7, 56.2, 7.7, 4Ø
C,3H,SNO3 {MW = 233.27); mass spectroscopy (MH+) N/A.
Example II-H
Synthesis of
N-(2-Biphenyl)-D,L-aianine
2-Aminobiphenyl (2 g, 11.8 mmol, Aldrich), triethylamine (i.2 eq.) and
ethyl 2-bromopropionate ( 1.1 eq., Aldrich) were combined and heated to
85°C
with stirring. After 7 days, the mixture was diluted with chloroform and
washed
with water. The organic portion was dried and concentrated to yield an oiI
which was purified by silica gel chromatography ( 1:1 CH,Ch/hexanes}. The
resulting oil was dissolved in a 1:2 mixture of water/dioxane (200 mL) and
LiOH (2 eq.) was added. After 2 hours, the mixture was concentrated to yield
an oil which was dissolved in water. The aqueous solution was washed with
ether then was adjusted to pH 3 with 5N HCl and extracted with ethyl acetate.
The organic portion was dried and concentrated to yield an oil which was
purified by silica gel chromatography (EtOAc) to yield the title compound.
Example II-I
Synthesis of
N-(Phenyl-furazan-3-yl)-D,L-alanine
._......~-.~.....,,....... ~
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4-Phenyl-furazan-3-ylamine (Maybridge) and ethyl pyruvate (Aldrich) were
dissolved in dry ethanol. Platinum on sulfide carbon (5%) was added and the
resulting mixture was hydrogenated ( 1000 psi, H,) at 150°C for 8
hours. The
reaction mixture was then filtered through Celite and the filtrate was
concentrated under reduced pressure. The residue was purified by silica gel
chromatography using CHCi3 as the eluent to provide the title compound as its
ethyl ester. The ethyl ester was then hydrolyzed using General Procedure II-A,
Method B (LiOH/H~0/dioxane) to provide the title compound.
Example .II-L
Synthesis of
S-(+)-3,5-Difluoromandelic Acid
Step A - Preparation of Methyl S-(t)-3 5-difluoromandelate
To a solution of 3,5-difluorobenzaldehyde (Aidrich) in Ct-I,C1, ( 100 mL)
was added ZnCI, (6.7 g, 21.1 mmol) to form a slurry. Trimethysilyl cyanide
(21.0 g, 211.2 mmol) dissolved in CH,C1~ (100 mL) was slowly added to the
slurry at 0°C. The resulting solution was stirred at room temperature
for 4 h.
The reaction mixture was then diluted with water and the organic layer
separated. The combined organic layers were concentrated to a residue. The
residue was dissolved with MeOH (200 mL) at 0°C and anhydrous HCl gas
bubbled into the solution for 10 min. After stirring at room temperature for
18
h, the solution was concentrated to a solid. The solid was dissolved in CH,Ch
/
H20 and the aqueous portion extracted with CH~CI,. The combined organics
were washed with brine, dried over anhydrous MgS04 and concentrated to a
solid (37.4 g, 87.6%), mp = 77-78°C.
'H NMR (300 MHz, CDC13): 8 = 6.97 {dd, J = 9.6 Hz, J = 1.79 Hz, 2H),
6.74 (dt, J = 8.82, J = 2.28 Hz, 1 H), 5.14 (d, J = 4.64 Hz, 1 H), 3.78 (s,
3H),
3.54 {d, J = 5.1 Hz, 1 H).
Step B - Preparation of Methyl S-(+)-3 5-difluoromandelate
Methyl (~)-3,5-difluoromandelate was separated via preparative chiral HPLC
to give a white solid having a melting point of 70-71 °C.
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C9HgF~03 (MW = 202.17); mass spectroscopy found (M+NH~+) 220Ø
Anal. calcd for C9H~F,03: C, 53.47; H, 3.99. Found: C, 53.40; H, 3.89.
Step C - Preparation of S-f+)-3,5-Difluoromandelic acid
A solution of methyl S-(+)-3,~-difluoromandelate ( 1 eq.) in 74% aqueous
THF was cooled to 0 °C and treated with lithium hydroxide. After 40
minutes
at 0 °C the reaction was complete by TLC. The contents were transferred
to a
separatory funnel and partitioned between CH,CI, and saturated aqueous
NaHCO;. The aqueous layer was acidified with 0.5 N NaHSOa and extracted
thrice with ethyl acetate. The combined extracts were washed with brine, dried
over Na,SO~,, filtered, and concentrated to a white solid having a melting
point
of 119-122 °C. The 'H NMR was consistent with known 3,5-
difluoromandelic
acid.
Example II-M
Synthesis of
2-Azido-(3,5-difluorophenyl)acetic Acid
Step A: To a three-necked flask equipped with a mechanical stirrer and a
nitrogen inlet tube was added 3,5-difluorophenylacetic acid (Aldrich) and THF.
The reaction mixture was cooled to -78°C and 1.2 eq. of
triethylarnine was
added, followed by dropwise addition of trimethylacetyl chloride ( 1.05 eq.)
(Aldrich). During the addition, the temperature was maintained at -
78°C. The
cold bath was then removed and replaced with an ice bath. The temperature
was allowed to warm to 0°C and stirring was continued for 1 hour. The
reaction mixture was then re-cooled to -78°C. To a second flask charged
with
THF, triphenylmethane (cat, 0.1 male %) and (S)-(-)-4-benzyl-2-oxazolidione
( 1.1 eq.) {Aldrich) at -78°C was added an n-butyllithium solution
dropwise until
an orange color persisted. This reaction mixture was stirred at -78°C
for 30
min. and then cannulated into the first reaction mixture. The resulting
mixture
was allowed to stir at -78°C for I hour and then quenched with 2.2 eq.
of acetic
acid. The solvent was removed under reduced pressure and the residue was
redissolved in dichloromethane and this solution washed with water, followed
by
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1 M potassium carbonate. The organic layer was then dried over sodium sulfate,
- filtered and concentrated. The residue was purified by LC 2000
' chromatography, eluting with EtOAC/Hexane ( 15:85). The resulting oil was
slurried in hexane to afford a white solid which was collected by filtration
to
give (S)-{-)-3-(3,5-difluorophenyacetyl)-4-benzyl-2-oxazolidione.
St- ~ B: To (S)-(-)-3-(3,5-difluorophenyacetyl)-4-benzyl-2-oxazolidione (3.0
mM) in 20 mL of dry THF cooled to -78°C was added LiHMDS {1.05 eq.)
dropwise while maintaining the temperature at -78°C. The reaction
mixture was
allowed to stir at -78°C for 15 min. and then a pre-cooled (-
60°C) solution of
trisyl azide ( 1.12 eq. ) in 10 mL of THF was added. The reaction mixture was
allowed to stir an additional 10 min. and then was quenched with 4.4 eq. of
acetic acid. Using a warm water bath, the temperature was raised to 30-
40°C
for 6 hrs. The reaction mixture was then poured into a separatory funnel and
extracted into dichloromethane. The organic Iayer was washed with bicarbonate
solution, followed by brine, and then dried over sodium sulfate, filtered and
solvent removed. The residue was purified by LC 2000 chromatography to
afford methyl 2-azido-2-(3,5-difluorophenyl)acetate.
Step C: To a solution of methyl 2-azido-2-(3,5-difluorophenyl)acetate in
THF/H,0 (2.6:1 ) cooled to 0°C was added 1.7 eq. of lithium
hydroxide. The
reaction mixture was stirred at room temperature for 3 hours and then poured
into a separatory funnel. The mixture was extracted into water and washed with
ether. The aqueous layer was acidified with 1N HCl and extracted with ethyl
acetate. The organic layer was then washed with water and brine. The organic
layer was dried over sodium sulfate, filtered and concentrated under reduced
pressure to give 2-azido-2-(3,5-difluorophenyl)acetic acid.
Example II-N
Synthesis of
(R)-N,N'-Di-BOC-2-Hydrazinopropionic Acid
Step A: To (S)-(-)-4-benzyl-2-oxazolidanone (Aldrich) in THF cooled to
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-50°C was added n-butyllithium 1.1 eq. ( 1.6 M in hexane) dropwise: The
reaction mixture was allowed to warm to -20°C and then was re-cooled to
-78°C
and propionyl chloride ( 1.1 eq) was added in one portion. The reaction
mixture
was allowed to stir an additional 15 min. at -78°C and then was allowed
to
warm to room temperature. The reaction was then quenched with a saturated
solution of sodium bicarbonate and extracted with ethyl acetate. The organic
extracts were washed with water, followed by brine and then dried over sodium
sulfate, filtered and concentrated to give (S)-(-)-3-propionyl-4-benzyl-2-
oxazolidanone.
Stet? B: To a solution of (S)-(-)-3-propionyl-4-benzyl-2-oxazolidanone in
THF at -78°C was added KHMDS ( 1.05 eq.) (Aldrich) dropwise. The
reaction
mixture was allowed to stir at -78°C for 30 min. and then a precooled
solution
of di-tert-butyl-azodicarboxylate (Aldrich) was added via a cannula. After 5
min. 2.6 eq. of acetic acid was added. The reaction mixture was then extracted
with dichloromethane and the organic layer was washed with 1 M potassium
phosphate. The organic layer was then dried over sodium sulfate, filtered and
concentrated to give (S)-(-)-3-[(R)-N,N'-di-BOC-2-hydrazinopropionyl]-4-
benzyl-2-oxazolidanone.
Step C: To {S)-(-)-3-[(R)-N,N'-di-BOC-2-hydrazinopropionyl]-4-benzyl-2-
oxazolidanone (0.49 moles) at 0°C in 8 mL of THF and 3 mL of water was
added LiOH {1.7 eq.) and H~OZ (3.0 eq.) and the reaction mixture was stirred
at
room temperature for 3 hours. The reaction mixture was then poured into a
seraratory funnel and diluted with water. The aqueous mixture was extracted
with ethyl acetate and then acidified to pH 2.0 with 1N HCl and extracted with
ethyl acetate. The organic layer was then dried over sodium sulfate, filtered
and
solvent removed to give (R)-N,N'-di-BOC-2-hydrazinopropionic acid which was
used without further purification.
Example II-O
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Synthesis of
3,5-Difluorophenyl-a-oxoacetic Acid
_ Step A: _ Ethyl 3,5-difluorophenyl-a-oxoacetate was prepared from 1-bromo-
3,5-difluorobenzene (Aldrich) according to the procedure described in J. Org.
Chem., 4~ ( 14), 2883-2887 ( 1980).
St_ ep B: Ethyl 3,5-difluorophenyl-a-oxoacetate was hydrolyzed using
General Procedure II-A (Method B) to afford 3,5-difluorophenyl-a-oxoacetic
acid.
Example II-P
Synthesis of
Cyclopentyl-a-hydroxyacetic Acid
The title compound (CAS No. 6053-71-0) was prepared in two steps from
cyclopentylmethanal (CAS No. 872-53-7, Wiley) using the procedure described
by Gibby, W. A.; Gubler, C. J. Biochemical Medicine 1982, 27, 15-25.
Example II-Q
Synthesis of N-(3,4-dichlorophenyl)alanine
Using the procedure set forth in U.S. Patent No. 3,598,859, the disclosure of
which is incorporated herein by reference in its entirety, N-(3,4-
dichlorophenyl)alanine was prepared. Specifically, to a solution of 3,4-
dichloroaniline ( 1 equivalent) (Aldrich) in isopropanol (about 500 mL per
mole
of 3,4-dichloroaniline) is added water (about 0.06 mL per mL of isopropanol)
and 2-chloropropionic acid (2 equivalents) (Aldrich). This mixture is warmed
to
40°C and sodium bicarbonate (0.25 equivalents) is added in successive
portions
before heating under reflux for 4-5 days. After cooling, the reaction mixture
is
poured into water and the unreacted 3,4-dichioroaniline is removed by
filtration.
The filtrate is acidified to pH 3-4 with concentrated hydrochloric acid and
the
resultant precipitate is filtered, washed and dried to yield the title
compound,
m.p. = 148-149°C.
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Example II-R
Synthesis of N-(3,5-difluorophenyl)alanine
Using tire procedure set forth in U.S. Patent No. 3,598,859 and Example Q
above, N-(3,5-difluorophenyl)alanine was prepared using 3,5-difluoroaniline
(Aldrich) and 2-chloropropionic acid (Aldrich).
Example II-S
Synthesis of
a-Fluoro-3,5-difluorophenylacetic Acid
Step A - Synthesis of Methyl 3,5-Difluoromandelate
To a solution of 3,5-difluoromandelic acid (Fluorochem) in methanol was
bubbled HCl gas for 10 minutes. The reaction was refluxed overnight. The
mixture was then concentrated in vacuo and the residue was taken up in ethyl
acetate and washed with saturated NaHC03 and brine. The organic layer was
dried over Na,S04, filtered, and concentrated to give the title intermediate
as a
white solid.
C9HgF,03 (MW=202.17); mass spectroscopy 202.
'H NMR (300 MHz, CDCl3): 8 = 7.00 (2H, d, J=6.58 Hz), 6.76 ( 1 H, t,
J=8.86 Hz), 5.16 (1H, d, J=5.29 Hz), 3.81 (3H, s), 3.54 (1H, d, J=5.39 Hz}.
Step B - Synthesis of MethyI a-Fiuoro-3,5-difluorophenylacetate
A solution of diethylaminosulfur trifluoride (DAST) ( 1.1 eq) in methylene
chloride was cooled to 0°C and a pre-cooled solution of methyl 3,5-
difluoromandelate ( 1 eq) in methyiene chloride was added. The transfer flask
was rinsed with a small portion of methylene chloride. After I S minutes, the
cooling bath was removed and the reaction mixture was stirred an additional 40
minutes at ambient temperature. The mixture was poured over ice and the
layers separated. The organic phase was washed with saturated NaHC03 and
brine. The organic layer was dried over Na,S04, filtered, and concentrated.
The residue was purified via HPLC eluting with 7% ethyl acetate/hexanes
providing the title intermediate as a yellow oil.
C9H,F302 (MW=204.16); mass spectroscopy 204.
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Anal. calcd for C9H.,F30,: C, 52.95; H, 3.46. Found: C, 52.8U; H, 3.73
- St. ep C = Synthesis of a-Fluoro-3,5-difluorophenylacetic Acid
Following General Procedure II-A, Method B and using methyl a-fluoro-
3,5-difluorophenylacetate, the title intermediate was prepared as a white
solid
having a melting point of 100-102°C.
C$HSF30, (MW = 190.13); mass spectroscopy 190.
Anal. calcd for C8HSF30z: C, 50.54; H, 2.65. Found: C, 50.47; H, 2.79.
The following Examples 1-20 illustrate the synthesis of compounds of the
present invention.
Example 1
Synthesis of
(S)-5-[1-N [N (3,5-Difluorophenylacetyl)-L-alaninyl]amino-
2-phenylethyl]-3-methyl-1,2,4-oxadiazole
Following General Procedure A above and using N-(3,5-
difluorophenylacetyl)-L-alanine (Example II-B) and (S)-5-( 1-amino-2-
phenylethyl)-3-methyl-1,2,4-oxadiazole hydrochloride (Example B), the title
compound was prepared as a solid having a melting point of 159-162°C.
The
reaction was monitored by tlc (Rf = 0.6 in 10% MeOH/CHC13) and the product
was purified by silica gel chromatography using 7% MeOH/CHCI3 as the eluent,
followed by recrystallization from 1-chlorobutane.
NMR data was as follows:
'H-nmr (DMSO-db): 8 = 4.30 (m, 1H), 5.26 (m, 1H).
C,,H"F,N403 (MW = 428.44); mass spectroscopy (MH+) 428.
Example 2
- Synthesis of
(S)-2-[1-{3,5-Difluorophenylacetamido)ethyl]-4-ethoxycarbonyl-2-thiazoiine
Following General Procedure A above and using 3,5-difluorophenylacetic
acid (Aldrich) and (S)-2-( 1-aminoethyl)-4-ethoxycarbonyl-2-thiazoline
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hydrochloride (Example G), the title compound was prepared as a semisolid.
The reaction was monitored by tlc (Rf = 0.2 in 1:1 EtOAc/hexanes) and the
product was purified by silica gel chromatography using 1:1 EtOAc/hexanes as
the eluent.
NMR data was as follows:
'H-nmr (CDCl3): 8 = 1.33 (t, 3H), 1.44 (t, 3H), 3.55 (s, 2H), 3.60 {m, 2H),
4.37 (m, 2H), 4.85 (m, 1 H), 5.05 (m, 1 H), 6.46 (t, 1 H), 6.74 (t, 1 H), 6.84
(d,
2H).
C,6H,8F,N~03S (MW = 356.39}; mass spectroscopy (MH~) 356.
Example 3
Synthesis of
1-tert-Butoxycarbonyl-2-[1-(N carbobenzyloxy)aminoethyl]
4-methoxycarbonyl-4-phenylmethyl-2-imidazoline
To a 1:1 mixture of the product from Example J, Step C ( 1.815 g, 1 eq.) in
THF (9 mL)/H~O (9 mL) at ambient temperature was added NaHC03 (0.377 g,
1.50 eq.) and di-tert-butyl dicarbonate (1.304 .g, 2.00 eq.) (Aldrich) in THF
(6
mL). The resulting pale yellow mixture was stirred at ambient temperature for
1
hour and then additional NaHC03 (0.188 g, 0.75 eq.) and di-tert-butyl
dicarbonate (0.652 g, 1.00 eq.) in THF (3 mL) were added and the mixture was
stirred for an additional hour. The mixture was then diluted with ethyl
acetate,
washed with brine (2x), dried over Na,S04, filtered, concentrated in vacuo,
and
purified by flash column chromatography casing 3:2 hexanes/EtOAc as the eluent
to provide the title compound in 77% yield as a viscous oil (Rf = 0.31 in 3:2
hexanes/EtOAc). The racemic 1:1 mixture of diastereomers was inseparable by
flash chromatography.
NMR data was as follows:
H-nmr (DMSO-d3, 250 MHz): 8 = 7.38-7.09 (m, 1 OH), 6.00 (bd, 1 H, J=
7.75 Hz), 5.21-5.05 (m, 3H), 4.15 (d, 1 H, J= 11.76 Hz), 4.08 (d, 1 H, J=
11.26
Hz), 3.85-3.80 (m, 2H), 3.78 (s, 3H), 3.20-3.00 (m, 2H), 1.43 (s, 9H), 1.36
(d,
1.SH, J= 6.75 Hz), 1.24 (d, 1.SH, J= 6.75 Hz).
Cz,H33N3Ob (MW = 495.58); mass spectroscopy (MH+) 496.4.
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Example 4 -
Synthesis of
. 1-tent-Butoxycarbonyl-2-[1-(3,5-difluorophenylacetamido)ethylJ-
4-methoxycarbonyl-4-phenylmethyl-2-imidazoline
Following General Procedure C above and using 3,5-difluorophenylacetic
acid (Aldrich) and I -tort-butoxycarbonyl-2-{ 1-aminoethyl}-4-methoxycarbonyl-
4-
phenylmethyl-2-imidazoline (Example J), the title compound was prepared as an
amorphous solid. The reaction was monitored by tlc (Rf = 0.34 in 2:1
hexanes/EtOAc) and the product was purified by flash column chromatography.
NMR data was as follows:
'H-nmr (CDC13, 250 MHz): 8 = 7.28-7.20 (m, 3H), 7.16-7.12 (m, I H),
7.07-7.03 (m, 1 H), 6.91-6.80 (m, 3H), 6.77-6.67 (m, 1 H), 5.35-5.21 (m, 1 H),
4.17-4.05 (m, 1 H}, 3.86-3.79 (m, 1 H), 3.79 (s, 3H), 3.55 (s, 1 H), 3.52 (s,
1 H),
3.18-2.99 (m, 2H), 1.43 (s, 4.5H), 1.41 (s, 4.5H), 1.33 (d, 1.5H, J= 6.75 Hz),
1.21 (d, 1.5H, J= 6.75 Hz).
C,,H3,F,N~05 (MW = 515.56); mass spectroscopy (MHt) 516.2.
Example 5
Synthesis of
2-[ 1-(3,5-Difluorophenylacetamido)ethylJ-
4-methoxycarbonyl-4-phenylmethyl-2-imidazoline
Following General Procedure D above and using the product from Example
4 above, the title compound was prepared as a white solid (93%) having a
melting point of 158-160°C. The reaction was monitored by tlc (Rf =
0.31 in
95:5 DCM/MeOH).
NMR data was as follows:
'H-nmr (DMSO-db and CDC13, 250 MHz): 8 = 8.09 (bt, IH, J= 7.75 Hz),
7.28-7.19 (m, 3H), 7.14-7.09 (m, 2H}, 6.89 (bd, 2H, J= 7.25 Hz), 6.74 (bt, 1
H,
J= 9.13 Hz), 4.62-4.48 (m, 1 H), 3.88 {bd, 1 H, J= 12.51 Hz), 3.68 (s, 3H),
3.6I-
3.54 (m, 1 H), 3.50 (s, 2H), 3.11-2.92 (m, 2H), 1.31 (d, 1.SH, J= 6.75 Hz),
1.30
(d, 1.5H, J= 7.00 Hz).
C"H,°F,N303 (MW = 415.44); mass spectroscopy (MH+) 416.1.
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Example 6 -
Synthesis of
-- (S)-2-[1-(3,5-Difluorophenylacetamido)ethyl]-
5(R,S)-ethoxycarbonyl-2-oxazoline
Following General Procedure A and using 3,5-difluorophenylacetic acid
(Aldrich} and (S)-2-(1-aminoethyI)-5(R,S)-ethoxycarbonyl-2-oxazoline (Example
F), the title compound was prepared as a semisolid. The reaction was monitored
by tlc (Rf = 0.3 in 5% MeOH/CHC13) and the product was purified by silica gel
chromatography using 5% MeOHICHC13 as the eluent.
NMR data was as follows:
'H-nmr (DMSO-db): 8 = 1.20 (t, 3H}, 1.30 (d, 3H), 3.50 (s, 2H), 3.80 {m,
1 H), 4.05 (m, 1 H), 4.1 S (q, 2H), 4.55 (m; 1 H), 5.10 (m, 1 H}, 6.97 (d,
2H), 7.08
(m, 1 H), 8.60 {d, 1 H).
C,6H,gF,N,04 (MW = 340.33); mass spectroscopy (MH+) 340.
Example 7
Synthesis of
2-[ 1-(3,5-Difluorophenylacetamido)ethyl]-
4-methoxycarbonyl-4-phenyl-2-imidazoline
Step A -- Synthesis of 1-tert-Butoxycarbonyl-2-[1-(3,5-
difluorophenylacetamido)ethyl]-4-methoxycarbonyl-4-phenyl-2-imidazoline
Following General Procedure C above and using 3,5-difluorophenylacetic
acid (Aldrich) and 1-tert-butoxycarbonyl-2-(1-aminoethyl)-4-methoxycarbonyl-4-
phenyl-2-imidazoline (Example K), the title compound was prepared as a white
foam (90%). The reaction was monitored by tlc (Rf = 0.39 in 3:2
hexanes/EtOAc) and the product was purified by flash column chromatography.
NMR data was as follows:
'H-nmr (CDCl3, 250 MHz): 8 = 7.38-7.31. (m, SH), 7.09 (d, 1H, J= 7.50
Hz), 6.90-6.82 (m, 21-1), 6.76-6.66 (m, 1 H), 5.57 (p, 1 H, J= 7.50 Hz), 4.83
(d,
1 H, J= 11.26 Hz}, 3.87 (d, 1 H, J= 11.26 Hz), 3.72 (s, 3H), 3.56 (s, 2H),
1.49 (s,
9H), 1.48 (d, 3H, J= 7.00 Hz).
CZ6H,9FZN3O5 (MW = 501.53); mass spectroscopy (MH+) 502.
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Step B -- Synthesis of 2-[1-(3,5-Difluorophenylacetamido)ethyl]-
4-methoxycarbonyl-4-phenyl-2-imidazoline
Following General Procedure D above and using the product from Step A
above, the title compound was prepared as a white foam (89%), The reaction
was monitored by tlc (Rf = 0.24 in 95:5 DCM/MeOH) and the product was
purified by flash column chromatography.
NMR data was as follows:
'H-nmr (DMSO-db, 300 MHz): 8 = 8.47 {d, 1 H, J= 7.89 Hz), 7.34-7.27 (m,
SH), 7.12-7.06 (m, 1 H), 7.03-7.00 (m, 2H), 4.55 (p, 1 H, J= 7.35 Hz), 4.34
(bm,
1H), 3.62 (s, 2H), 3.53 {s, 2H), 1.31 (d, 3H, J= 7.00 Hz)..
C2,HZ,F~N303 (MW = 401.41); mass spectroscopy (Ml-i+) 402.
Example 8
Synthesis of 1-tert-Butoxycarbonyl-
2-(3,5-difluorophenylmethyl)-4-ethoxycarbonyl-4-methyl-2-imidazoline
Step A -- Synthesis of Ethyl 2,3-Diamino-2-methylpropionate
Following the procedures described in Gilbert, et al., Tetrahedron, 1995 SI ,
6315-6336, the title compound was prepared as an oil. Purification was by
Kugelrohr distillation.
NMR data was as follows:
'H-nmr (DMSO-d~): 8 = 4.06 (m, 2H), 2.70 (d, 1H), 2.46 (d, 1H), 2.18 (bs,
4H), 1.18 (t, 3H), 1.10 (s, 3H).
C6H,4N202 (MW = 146.19); mass spectroscopy (MH+) 147.
Step B -- Synthesis of 2-(3,S-Difluorophenylmethyl)-4-ethoxycarbonyl-4-
methyl-2-imidazoline
In a round bottom flask fitted with an addition funnel was added 2.25 g
(9.55 mmole) of the product from Example L -- Step A above and 20 mL of
ethanol. The flask was cooled to 0° C and stirring was initiated under
an
atmosphere of nitrogen. The product from Step A above ( 1.40 g, 9.55 mmole)
. in 20 ml of ethanol was slowly added to the reaction via an addition funnel.
The reaction was allowed to slowly warm to room temperature and stir
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overnight. The mixture was then filtered through a sintered glass funnel and
the
filtrate was concentrated on the rotary evaporator. The residue was
partitioned
between 50 i'nL of 1.0 N sodium hydroxide and 50 mL of dichloromethane.
The organic phase was separated and the aqueous layer was washed twice with
50 mL of dichloromethane. The combined organic layers were washed with 50
mL of saturated brine and dried over sodium sulfate. Filtration followed by
concentration on the rotary evaporator, yielded a yellow oil which was
purified
via flash chromatography on silica gel 60 (230-400 mesh) using 95:5
DCM/MeOH as the eluent. Concentration of the fractions gave 2.61 g (82%
yield) of the title compound (Rf = 0.17 in 95:5 DCM/MeOH).
NMR data was as follows:
'H-nmr (DMSO-db): 8 = 7.05 (m, 3H), 4.04 {m, 2H), 3.75 {d, J = 11.71 Hz,
1H), 3.49 (s, 2H), 3.27 (d, J = 11.73 Hz, 1H), 1.29 (s, 3H), 1.17 (t, 3H).
C,4H,6F,N20~ {MW = 282.29); mass spectroscopy (MH~) 282.1.
Step C -- Synthesis of 1-tert-Butoxycarbonyl-
2-(3,5-difluorophenylmethyl)-4-ethoxycarbonyl-4-methyl-Z-imidazoline
In a round bottom flask fitted with an addition funnel was added the
product from Step B above (2.00 g, 7.08 mmole}, 0.743 g (8.85 mmole) of
sodium bicarbonate, 30 mL of water, and 30 mL of THF. Stirring was initiated
under an atmosphere of nitrogen and di-tert-butyl dicarbonate (3.47 g, 15.90
mmole) (Aldrich) in 15 mL of THF was added to the reaction via the addition
funnel. After stirring for 1 hour, an additional 627 mg (7.05 mmole) of sodium
bicarbonate and 1.I7 g (5.40 mmole) of di-tert-butyl dicarbonate were added to
the reaction. The reaction was monitored using thin layer chromatography, and
when the reaction was complete, the reaction mixture was partitioned between
250 mL of ethyl acetate and 250 mL of saturated brine. The organic layer was
washed twice with a 250 mL of saturated brine and then dried over sodium
sulfate. Filtration followed by concentration on the rotary evaporator yielded
a
yellow oil which was purified by flash chromatography on silica gel 60 (230-
400 mesh) using 15:85 ethyl acetate/dichloromethane as the eluent.
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Concentration of the fractions gave 2.10 g (78 % yield) of the title compound
(Rf = 0.15 in 15:85 EtOAc/hexanes).
NMK data was as follows:
'H-nmr (DMSO-db): b = 7.07 (m, 1H), 6.94 (m, 2H), 4.05 (m, SH), 3.57
(d, 1H), 1.38 (d, 12H}, 1.19 (t, 3H},
'3C-nmr (DMSO-db): 8 = 172.116, 163.599, 160.348, 157.624, 149.352,
140.855, 111.511, 101.584, 81.677, 69.950, 60.758, 55.442, 34.855, 27.423,
24.203, 13.581.
C,9H,4FZN,0~ (MW = 382.41 ); mass spectroscopy (MH') 382. I .
Example 9
Synthesis of
2-[1-N [N (3,5-Difluorophenylacetyl)
L-alaninyl]amino-1-phenyl] methyl-2-thiazoline
Following General Procedure F above and using N-( 1-cyano-1-
phenylmethyl)-N'-(3,5-difluorophenylacetyl)-L-alaninamide and 2-
mercaptoethylamine, the title compound was prepared. The reaction was
monitored by tlc (Rf = 0.3 in 5% MeOH/CHC13) and the product was purified
by silica gel chromatography using 5% MeOH/CHC13 as the eluent.
NMR data was as follows:
'I-I-nmr (CDC13): b = 1.31 (m, 3H), 3.31 (m, 2H), 3.50 (d, 2H), 4.24 (m,
2H), 4.55 (m, 1 H), 4.64 (d, 1 H), 6.30 (m, I H), 6.76 (m, 2H), 7.33 (d, SH),
7.52
(m, 1 H).
C,,HZ,FZN30,S (MW = 417.48); mass spectroscopy (MH+) 417.
Example 10
Synthesis of
2-[ 1-(3,5-Difluorophenylacetamido)-1-phenyl] methyl
4-ethoxycarbonyl-2-thiazoiine
Following General Procedure G above and using N-(3,5-
difluorophenylacetyl)phenylglycinonitrile from Example I above and L-cysteine
ethyl ester hydrochloride (Aldrich), the title compound was prepared as an
amorphous solid. The reaction was monitored by tlc (Rf = 0.5 in 1:1
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EtOAc/hexanes) and the product was purified by silica gel chromatography
using 40% EtOAc/hexanes as the eluent.
NMR-data was as follows:
'H-nmr (CDC13): b = 1.36 (t, 3H), 3.6 (m, 4H), 4.30 (m; 2H), 5.10 and
5.20 (t, 1 H}, 5.84 (d, 1 H), 6.72 (m, 1 H), 6.84 (m, 2H), 7.38 (m, SH).
C,,H,oF,N~03S (MW = 418.47); mass spectroscopy (MH~) 418.
Example 11
Synthesis of
2-((S)-1-(3,5-dichloroanilino)ethyl]-{S)-4-methoxycarbonyl-2-oxazolidine
Following General Procedure H above, the title compound was prepared
as an oil. The reaction was monitored by tlc (Rf = 0.~ in 3:2 hexanes/EtOAc)
and the product was purified by preparative tlc chromatography using 3:2
hexanes/EtOAc as the eluent.
NMR data was as follows:
'H-nmr (CDCl3): 8 = 1.5 (d, J = 7 Hz, 3H}, 3.79 (s, 3H), 4.15-4.3 (m,
1 H), 4.3-4.65 (m, 3H), 4.7-4.85 (m, 1 H), 6.45 (s, 2H), 6.7 (s, I H).
r3C-nmr (CDC13): b = 14.2, 42.08, 42.15, 47.9, 62.9, 65.3, 106.67,
106.72, 113.0, 130.7, 143.3. 166.3.
C,3H,~Cl~N~03 (MW = 317); mass spectroscopy (MH+) NIA.
Example 12
Synthesis of
(S)-S-[1-N [N (3,5-Difluorophenylacetyl)-L-alaninyl]amino-
1-phenyl]methyl-3-methyl-1,2,4-oxadiazole
Following General Procedure A above and using N (3,5-
difluorophenylacetyl)-L-alanine (Example II-B) and (S}-S-( I -amino- I -
phenylmethyl)-3-methyl-1,2,4-oxadiazole hydrochloride (Example C), the title
compound was prepared as a solid (3:2 mixture of diastereomers). The reaction
was monitored by tlc (Rf = 0.6 in 7% MeOH/CHC13) and the product was
purified by silica gel chromatography using 7% MeOH/CHC13, followed by
recrystallization from 1-chlorobutane/acetonitrile.
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NMR data was as follows: -
'H-nmr (DMSO-db): b = 1.21 (d, 3H), 2.32 (s, 3H), 3.53 {s, 2H), 4.43
' (m, 1 H), 6.35 (d, 1 H), 6.97 (d, 2H), 7.07 (m, 1 H), 7.38 (broad s, SH),
8.38 (d,
1 H), 9.27 (d, 1 H). _
S C,°H,°F~N403 (MW = 414.41 ); mass spectroscopy (MH~) 414.
Example 13
Synthesis of
(S)-5-[1-N [lV (3,5-Difluorophenylacetyl)-L-alaninyl)amino-
1-phenyl)methyl-3-phenyl-1,2,4-oxadiazole
Following General Procedure A above and using N-(3,5-
difluorophenylacetyl)-L-aianine (Example II-B) and (S)-5-(1-amino-1-
phenylmethyl)-3-phenyl-1,2,4-oxadiazole hydrochloride (Example D), the title
compound was prepared. The reaction was monitored by tlc (Rf = 0.4 in 5%
MeOH/CHCl3) and the product was purified by silica gei chromatography using
5% MeOH/CHCI3 as the eluent.
NMR data was as follows:
'H-nmr (DMSO-db): 8 = 1.31 (d, 3I-1), 3.51 (s, 2I-1), 4.47 {m, 1H), 6.42
(d, IH), 6.97 (d, 2H), 7.07 (m, 1H), 7.35-7.60 (m, 8H), 7.97 (d, 2H), 8.40 (d,
1 H), 9.27 (d, 1 H).
C,6H"FZN403 (MW = 476.49); mass spectroscopy (MH~) 476.
Example l4
Synthesis of
(S)-5-[1-N (N (3,5-Difluorophenylacetyl)-L-alaninyl)amino-
1-phenyl]methyl-3-(4-methaxyphenylmethyl)-1,2,4-oxadiazole
Following General Procedure A above and using N-(3,5-
difluorophenylacetyl)-L-alanine (Example II-B) and (S )-5-( 1-amino-1-
phenylmethyl)-3-(4-methoxyphenylmethyl)- I ,2,4-oxadiazole hydrochloride
(Example E), the title compound was prepared ( I :2 mixture of diastereomers).
The reaction was monitored by tlc (Rf = 0.25 in 3.5% MeOH/CHCl3) and the
product was purified by silica gel chromatography using 3.5% MeOH/CHC13) as
the eluent.
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NMR data was as follows: - -
'H-nmr (DMSO-d3): 8 = 1.22 (d, 3H), 3.49 (s, 2H), 3.72 (s, 3H}, 3.98 (s,
2H), 4.42 (rrl, 1 H), 6.28 (d, 1 H), 6.86 {d, 2H), 6.96 (d, 2H), 7.07 (m, 1
H), 7.18
(d, 2H), 7.38 (broad s, 5H), 8.36 {d, 1 H), 9.18 (d, 1 H).
C,gHz6F2N4O4 (MW = 520.54); mass spectroscopy (MH') 520.
Example I S
Synthesis of
(4R)-4-[N (1S)-(1-Methoxycarbonyl-1-phenyl)methyl[carbamoyl-
2-(3,5-difluorophenyimethyl)-4-methyl-2-thiazoline
To a solution of (4R)-4-carboxy-2-(3,5-difluorophenylmethyl)-4-methyl-
2-thiazoline (0.2509 g, 1.00 eq.) from Example L above in THF (20 mL) at
0°C
under an atmosphere of nitrogen was added (S)-{+)-2-phenylglycine methyl ester
hydrochloride (0.2052 g, 1.10 eq.) {Aldrich), I-hydroxybenzotriazole hydrate
{0.1437 g, 1.15 eq.) (Aldrich), N,N diisopropylethylamine (0.371 mL, 2.30
eq.),
followed by I -{3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride
(0.2039 g, 1.15 eq.) (Aldrich). The cooling bath was removed and the mixture
allowed to warm to ambient temperature with stirring for I9 hours. The
solution was diluted with ethyl acetate, washed with 0.5 M aqueous HCl (2x),
di lute aqueous NaHC03 ( 1 x), and brine ( I x); then the organic phase was
dried
over Na~S04, filtered, concentrated in vacuo, and the residue purified by
flash
column chromatography using 2:1 hexanes/EtOAc as the eluent to yield the title
compound as a clear colorless viscous oil (0.3465 g, 90%).
NMR data was as follows:
'H-nmr {CDCIj, 300 MHz): b = 7.59 (d, 1H, J= 7.25 Hz), 7.36-7.27 (m,
SH), 6.80-6.78 (m, 2H), 6.75-6.68 (m, I H), 5.51 {d, I H, J= 7.37 Hz), 3.78
(s,
2H), 3.74 (s, 3H), 2.63 (d, I H, J= 11.59 Hz), 2.22 (d, 1 H, J= 11.60 Hz),
1.55 (s,
3H}.
Optical Rotation: [a],o = 65.3 (c 1.0, CHCl3):
C~,H2oF2Nz03S (MW = 418.47); mass spectroscopy (MH~) 418.3.
Example I6
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Synthesis of -
- 4-[N (S)-(1-Methoxycarbonyl-1-phenyl)methyl]carbamoyl-
2-(3,5-difluorophenylmethyl)-2-thiazoline
Following General Procedure A above and using L-phenylglycine methyl
ester and 2-(3,5-difluorophenylmethyl)-4-carboxy-2-thiazoline from Example H
above, the title compound was prepared as two separate diastereomeric isomers.
The reaction was monitored by tlc (Rf = 0.6 and 0.4 in 1:1 EtOAc/hexanes) and
the products were purified by silica gel column chromatography using 40%
EtOAc/hexanes as the eluent.
First Diastereomeric Isomer (amorphous solid):
NMR data was as follows:
'H-nmr (CDC13): d = 3.58-3.82 (m, 5H), 3.94 (s, 2H), 5.10 (t, 1H), 5.57
(d, 1 H), 6.75 (m, 1 H), 6.90 (d, 2H), 7.35 f m, 5H), 7.84 (d, 1 H).
C,9H,8F,N,03S {MW = 404.44); mass spectroscopy (MH+) NIA.
Second Diastereomeric Isomer (oil):
NMR data was as follows:
'H-nmr (CDC13): 8 = 3.61 (d, 2H), 3.75 (s, 3H), 3.84 (s, 2H), 5.13 (t,
1H), 5.56 (d, 1H), 6.72 (m, 1H), 6.80 (d, 2H), 7.33 (m, 5H), 7.66 (d, IH).
C,9H,8F~N,03S (MW = 404.44); mass spectroscopy (MH+) 404.
Example 17
Synthesis of
(S)-5-[1-N [N (3,5-Difluorophenylacetyl)-L-alaninyl]amino]ethyl-
3-ethyl-I,2,4-oxadiazole
Following General Procedure A and using N (3,5-difluorophenylacetyl)-
L-alanine (Example II-B) and (S)-5-(1-aminoethyl)-3-ethyl-1,2,4-oxadiazole
hydrochloride (Example A), the title compound was prepared as a solid having a
melting point of 181-183 °C. The reaction was monitored by tlc (Rf =
0.3 in 7%
MeOH/CHC13) and the product was purified by silica gel column
chromatography using 7% MeOH/CHC13 as the eluent, followed by
recrystallization from 1-chlorobutane.
NMR data was as follows:
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'H-nmr (DMSO-db): 8 = 1.20 (m, 6H), 1.45 (d, 3H), 2.70 (q; 2H),- 3.51
(s, 2H), 4.32 (m, 1 H), 5.13 (m, 1 H), 6.98 (d, 2H), 7.10 (m, 1 H), 8.3 8 (d,
I H),
8.73 (d, 1 H).'
Optical Rotation: [a~2o = +53.1 @ 589 nm (c 1.08, DMSO).
C"H,oF,N403 (MW = 366.37); mass spectroscopy (MH+) 367.
Example 18
Synthesis of
2-[ 1-(3,5-Difluorophenylacetamido)-1-phenyl] methyl-
(4R)-4-methoxycarbonyl-2-thiazoline
Following General Procedure G above and using N-(3,5-
difluorophenylacetyl)phenylglycinonitrile from Example I above and (R)
cysteine methyl ester hydrochloride (CAS No. 2485-62-3), the title compound
was prepared. The product was purified by silica gel chromatography using 1:1
EtOAc/hexanes as the eluent, followed by crystallization from 1-chlorobutane.
First Diastereomeric Isomer (higher Rf):
NMR data was as follows:
' H-nmr (CDC13): 8 = 5.08 {t, 1 H), 5. I 8 (t, 1 H), 5.47 (t, 1 H), 5.82 (m,
1 H).
C~oH,gF,N,03S (MW = 404.44); mass spectroscopy (M+) 404.
Second Diastereomeric Isomer (lower Rr}:
NMR data was as follows:
'H-nmr (CDC13): b = 5.12 (t, 1 H), 5.21 (t, I H), 5.47 (t, 1 H), 5.83 (m,
1 H).
C,oH,BFzN~03S (MW = 404.44}; mass spectroscopy (M+) 404.
Example 19
Synthesis of
2-[ 1-(3,5-Difluorophenyiacetamido)-1-phenyl] methyl-
(4S)-4-methoxycarbonyl-2-thiazoline
Following General Procedure G above and using N-(3,5-
difluorophenylacetyl)phenylglycinonitrile from Example I above and {S)-
cysteine
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methyl ester hydrochloride (Aldrich), the title compound was prepared. The
product was purified by silica gel chromatography using 2:3 EtOAc/hexanes as
' the eluent, f611owed by crystallization from 1-chlorobutane/hexanes.
First Diastereomeric Isomer (higher Rf): -
NMR data was as follows:
'H-nmr (CDjOD): b = 3.76 (s, 3H), 5.20 (t, 1H), 5.85 (s, 1H).
C~~H,gFZN,03S (MW = 404.44); mass spectroscopy (M+) 404.
Second Diastereomeric Isomer (lower Rf):
NMR data was as follows:
'H-nmr (CD30D): 8 = 3.78 (s, 3H), 5.22 (t, 1H), 5.83 (s, 1H).
C~oH,BF,N,O3S (MW = 404.44); mass spectroscopy (M+) 404.
Example 20
1S Synthesis of
N [2-(3,5-Difluorophenylmethyl)-4-methyl-2-imidazoline
4-carboxamido]-L-phenylglycine Methyl Ester
Step A -- Synthesis of 1-tert-Butoxycarbonyl-2-(3,5-
difluorophenylmethyl)-4-carboxy-4-methyl-2-imidazoiine
A round bottom flask containing a magnetic stir bar was charged with
150 mL of THF, 50 mL of a 0.1 N aqueous solution of lithium hydroxide, and 1-
tert-butoxycarbonyl-2-(3,S-difluorophenylmethyl)-4-ethoxycarbonyl-4-methyl-2-
imidazoline (Example 8, Step C) (2.58 g, 6.75 mmole). The reaction mixture
was stirred for 1 hour and then an additional 7S mL of a O.1N aqueous solution
of lithium hydroxide was added and stirring was continued for 3.S hours. The
reaction mixture was then concentrated in vacuo and the resulting residue was
partitioned between ethyl acetate and a 0.5N aqueous solution of hydrochloric
acid. The organic phase was separated and washed with an additional 0. SN
aqueous solution of hydrochloric acid. The organic phase was then dried over
Na~SO,~. The drying agent~was removed by filtration and the filtrate was
partially concentrated in vacuo. The resulting solid was isolated via vacuum
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filtration through a sintered glass funnel, followed by drying under vacuo at
80°C.
Mass spectroscopy data was as follows:
C,~HZONz04 (MW = 354.36); mass spectroscopy (MH+) 255.2.
Step B -- Synthesis of N [1-tert-Butoxycarbonyl-2-(3,5-
difluorophenylmethyl)-4-methyl-2-imidazoline-4-carboxamido]-L-
phenylglycine Methyl Ester
Following General Procedure A above and using (S)-(+)-2-phenylglycine
methyl ester hydrochloride and the product of Step A, the title compound was
prepared. The product was purified by flash silica gel chromatography (230-400
mesh) using 95:5 DCM/MeOH as the eluent.
First Diastereomeric Isomer (viscous oil):
NMR data was as follows:
'H-nnlr (CDC13): & = 7/60 (bs, 1H), 7.35 (s, 5H), 6.90 (m, 2H), 6.69 (m,
I H), 5.49 (d, 1 H), 4.12 (m, 3H), 3.72 (s, 3H), 3.61 (d, 1 H), 1.46 (s, 9H),
1.41
(s, 3H).
Second Diastereomeric Isomer (viscous oil):
NMR data was as follows:
'H-nmr (DMSO-db): 8 = 7.55 (bs, 1H), 7.31 (m, 5H), 6.84 (m, 2H), 6.67
(m, 1 H), 5.49 (d, 1 H), 4.08 (m, 3H), 3.72 (s, 3H), 3.56 (m, 1 H), 1.50 (s, 1
H),
1.43 (s, 9H).
Step C -- Synthesis of N [2-(3,5-Difluorophenylmethyl)-4-methyl-2-
imidazoline-4-carboxamido]-L-phenylglycine Methyl Ester
Following General Procedure D above and using the individual isomers
from Step B, the title compound was prepared as two separate isomers. These
products were purified by flash silica gel chromatography (230-400 mesh) using
97.5:2.5 DCM/MeOH as the eluent.
First Diastereomeric Isomer:
Mass spectroscopy data was as follows:
_....~_....._....,-....~,~,..", .._...._....,..... ....~". .........
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C,,H~,F,N~03S (MW = 401.42); mass spectroscopy (MI-I-) 402Ø
Second Diastereomeric Isomer:
Mass spectroscopy data was as follows: -
C,,HZ,F~N~03S (MW = 401.42); mass spectroscopy (MHT) 402Ø
Additionally, the following General Procedures and Examples provide
various carboxylic acids (and carboxylic acid esters which can be hydrolyzed
using General Procedures AC or BD below to afford the corresponding
carboxylic acids) which can be used to prepare additional compounds within the
scope of this invention.
GENERAL PROCEDURE AA
Reductive Amination
To a solution of the arylamine in ethanol in a hydrogenation flask was
added 1 equivalent of the 2-oxocarboxylic acid ester (e.g., pyruvate ester),
followed by 10% palladium on carbon (25 weight percent based on the
arylamine). The reaction was hydrogenated at 20 psi H, on a Parr shaker until
complete reaction was indicated by tlc (30 minutes to 16 hours). The reaction
mixture was then filtered through a pad of Celite 545 (available from Aldrich
Chemical Company, Inc.) and stripped free of solvent on a rotary evaporator.
The crude product residue was then further purified via chromatography.
GENERAL PROCEDURE AB
First Transesterification Technique
A solution of 1-~ equivalents of the desired alcohol was added to 1
equivalent of sodium hydride in toluene. After off gassing had ceased, the
compound to be transesterified, dissolved in toluene, was added. After 0.5
hours, the reaction was either heated to 40°C and placed under house
vacuum
(~20 mmHg), or nitrogen was bubbled through the solution while it was heated
at 90°C. The reaction was followed by tlc, and when the reaction was
complete
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the solution was cooled and quenched with water or 1 M HCI, and in smaller
scale reactions diluted with ethyl acetate. The organic phase was extracted
with
saturated aqueous NaHC03, then washed with saturated aqueous NaCI and dried
over MgSOa. The solution was stripped free of solvent on a rotary evaporator,
and the crude product residue was then further purified by chromatography.
Alternatively, the reaction mixture was worked-up by evaporation of the
solvents
and direct chromatography of the crude mixture.
This procedure is particularly useful in the case of costly and/or high
boiling alcohols.
GENERAL PROCEDURE AC
Second Transesterification Technique
The compound to be transesterified was placed in a large excess of the
desired alcohol. A catalytic amount of dry NaH was added, and the reaction
was followed by tlc until the presence of starting material was no longer
detected. The reaction was quenched with a few milliliters of 1 N HCI, and
after
a few minutes of stirring saturated aqueous NaHCO, was added. The organic
phase was washed with saturated aqueous NaCI and dried over MgSO,. The
solution was stripped free of solvent on a rotary evaporator, and the crude
product residue was then further purified by chromatography.
GENERAL PROCEDURE AD
Third Transesterification Techniq-ue
The compound to be transesterified was placed in a large excess of the
desired alcohol. A catalytic amount of dry NaH was added, and the reaction
was followed by tlc until the presence of starting material was no longer
detected. The reaction was quenched with a few milliliters of 1N HCI, and
after a few minutes of stirring saturated aqueous NaHCO, was added. The
volume of the reaction mixture was reduced on a rotary evaporator until the
excess alcohol was removed and then the remaining residue was taken up in
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ethyl acetate and additional water was added. The organic phase was washed
' with saturated aqueous NaCI and dried over MgSO,. The solution was stripped
free of solvent on a rotary evaporator, and the crude product residue was then
further purified by chromatography. -
- 5
This procedure is particularly employed in the case of low boiling,
inexpensive alcohols, miscible with water.
GENERALPROCEDURE AE
O-Alkvlation Technique
To a carboxylic acid compound (prepared, for example, by reductive
amination via General Procedure AA to provide for the N-aryl amino acid ester,
followed by hydrolysis via Procedure AF) in DMF was added 1.5 equivalents
K2C0" followed by 1 equivalent of alkylating agent (e.g., tert-butyl
bromoacetate). The reaction was stirred at room temperature for 2 hours, then
was quenched with water and extracted into ethyl acetate. The organic phase
was washed with saturated aqueous NaHCO" water, and saturated aqueous NaCI,
and was then dried over MgS04. The solution was stripped free of solvent on a
rotary evaporator to yield the crude product.
GENERALPROCEDURE AF
Ester H~lysis to Free Acid
To a carboxylic ester compound (prepared, for example, by reductive
amination via General Procedure AA to provide for the N aryl amino acid ester)
in a l :l mixture of CH,OH/H=O was added 2-5 equivalents of KzCO,. The
mixture was heated to 50°C for 0.5 to i .5 hours until tlc showed
complete
reaction. The reaction was cooled to room temperature and the methanol was
removed on a rotary evaporator. The pl-1 of the remaining aqueous solution was
adjusted to ~2, and ethyl acetate was added to extract the product. The
organic
phase was then washed with saturated aqueous NaCI and dried over MgSO~.
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The solution was stripped free of solvent on a rotary evaporator to yield the
crude product.
GENERALPROCEDURE AG -
N Heteroarylation of Alanine
A solution of 1.1 equivalents of L-alanine and 2 equivalents NaOH in
DMSO was stirred at room temperature for 1 hour, then 1 equivalent of 2-
chlorobenzothiazole was added. The mixture was heated to 100°C for 4
hours,
then cooled to room temperature and poured onto ice. The pH of the resulting
aqueous solution was adjusted to ~2, and the precipitated solid was removed by
filtration. This solid was then dissolved in 1 N NaOH and the resulting
solution
was filtered through a pad of Celite 545. The pH of the filtrate was adjusted
to
~2, and the white precipitate was removed by filtration and washed with water
to yield the crude product.
GENERAL PROCEDURE AH
EDC Coupling
To a 1:1 mixture of the desired acid and alcohol in CHzCIz at O°C
was
added 1.5 equivalents triethylamine, followed by 2.0 equivalents
hydroxybenzotriazole monohydrate, then 1.25 equivalents of ethyl-3-(3-
dimethylamino)-propyl carbodiimide'HCl (EDC). The reaction was stirred
overnight at room temperature, then transferred to a separatory funnel and
washed with water, saturated aqueous NaHCO" 1N HCI, and saturated aqueous
NaCI, and was then dried over MgSO,. The solution was stripped free of
solvent on a rotary evaporator to yield the crude product.
GENERAL PROCEDURE AI
Oxime or Amine Coupling Technique
The trichlorophenyl ester (1 eq) of a carboxylic acid was stirred in DMF
or THF. The oxime or amine ( 1.2 eq) was added and the mixture was stirred at
ambient temperature for 1-4 hours. In cases where the hydrochloride salt form
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of an amine was used, a suitable base such as N,N diisopropylethylamine ( 1.2
eq) was also added. The resulting mixture was concentrated under reduced
pressure to yield a crude product which was used without purification or was
purified by silica gel chromatography andior crystallization.
GENERAL PROCEDURE AJ
Alkylation Technigue
The amine ( 1 eq), the a-bromo ester ( 1.1 eq) and a suitable base (such as
triethylamine) (2 eq) were stirred in chloroform. The resulting solution was
heated at reflux for 4-12 hours. After cooling, the mixture was diluted with
chloroform and washed with water. The organic portion was dried (sodium
sulfate) and concentrated under reduced pressure. The crude product was
purified by silica gel chromatography.
GENERAL PROCEDURE AK
Oxime or Alcohol Coupling TechnicFue
The carboxylic acid ( 1 eq) was stirred in a suitable solvent (such as THF,
dioxane or DMF). An alcohol or oxime { 1-5 eq) was added. EDC
hydrochloride ( 1.2 eq) and hydroxybenzotriazole hydrate ( 1 eq) were added. A
suitable base (such as 4-methylmorpholine or triethylamine) {0-1 eq) was
added.
A catalytic amount (0.1 eq) of 4-dimethylaminopyridine was added. The
mixture was stirred at ambient temperature and under a dry atmosphere of
nitrogen. After 20 hours, the mixture was concentrated under reduced pressure.
The resulting concentrate was partitioned between ethyl acetate and water. The
organic portion was separated and washed with aqueous sodium bicarbonate and
brine. The organic portion was dried (sodium sulfate) and concentrated under
reduced pressure. The crude product was used without purification or was
purified by silica gel chromatography and/or crystallization.
GENERAL PROCEDURE AL
EDC Coupling
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The carboxylic acid was dissolved in methylene chloride. The amino
acid (1 eq.), N-methylmorpholine (5 eq.} and hydroxybenzotriazole monohydrate
( 1.2 eq.) were added in sequence. A cooling bath was applied to the round
bottomed flask until the solution reached 0°C. At that time, 1.2 eq. of
1-{3-
dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC) was added.
The solution was allowed to stir overnight and come to room temperature under
nitrogen pressure. The reaction mixture was worked up by washing the organic
phase with saturated aqueous sodium carbonate, 0.1 M citric acid, and brine
before drying with sodium sulfate. The solvents were then removed to yield
crude product. Pure products were obtained by flash chromatography in an
appropriate solvent.
GENERAL PROCEDURE AM
Triflate Displacement
To a 0°C solution of iso-butyl R-{+)-lactate in CHzCI~ was added
1.1
equivalents of trifluoromethanesulfonic anhydride. After stirring at room
temperature for 20 min, 1.1 equivalents of 2,6-lutidine was added and stirring
was continued for 10 min. This solution was then transferred to a flask
containing 1 equivalent the arylamine and 1 equivalent N,N
diisopropylethylamine in CHZCh or CH3N0., at 0°C. The reaction was held
overnight at room temperature and then stripped free of solvent on a rotary
evaporator. The residue was dissolved in ethyl acetate, washed with 5% citric
acid, followed by saturated aqueous NaCI, dried over magnesium sulfate or
sodium sulfate and then the solution was stripped free of solvent on a rotary
evaporator to yield the crude product, which was then purified by
chromatography.
GENERAL PROCEDURE AN
BOC Removal
The BOC-protected compound was added to a 1:1 mixture of CH,CI, and
trifluoroacetic acid, and was stirred until tlc indicated complete conversion,
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typically 2h. The solution was then stripped to dryness and the residue was
' taken up in ethyl acetate and extracted with dilute HCI. The acid reaction
was
neutralized and extracted with ethyl acetate. The organic phase was washed
with saturated aqueous NaCI and dried over MgS04. The solution was stripped
free of solvent on a rotary evaporator to yield the product.
GENERAL PROCEDURE AO
Synthesis of Pyruvate Esters
To a mixture of pyruvic acid (8.8 g, 0.1 mol) (Aldrich) in 100 mL of
benzene was added iso-butanol { 14.82 g, 0.2 mol) and a catalytic amount of
p-toluenesulfonic acid. The mixture was then refluxed using a Dean Stark
apparatus. After 4 hours, the reaction appeared to be complete with the
isolation
of 1.8 g {0.1 mol) of water. The benzene and iso-butanol were removed on a
rotary evaporator. The residue (14 g, 0.1 mol), which was primarily the
pyruvate iso-butyl ester by nmr ['H-Nmr (CDC13): 8 = 4.0 (d, 2H), 2.5 (s, 3H),
2.0 (m, 1 H), 1.0 (d, 6H)], was used without further purification. By
substituting
other alcohols in place of iso-butanol (e.g., ethanol, isopropanol, n-butanol,
benzyl alcohol and the like), other esters of pyruvic acid can be prepared in
a
similar manner.
GENERALPROCEDURE AP
Aromatic Nucleophilic Substitution of Fluorobenzenes
A mixture of 1.82 g ( 10 mmol) of D,L-alanine iso-butyl ester
hydrochloride, the fluorobenzene ( 10 mmol) and 3 g of anhydrous potassium
carbonate in 10 mL of DMSO was stirred at 120°C for 2-5 hours. The
reaction
mixture was then cooled to room temperature and diluted with 100 mL of ethyl
acetate. The ethyl acetate extract was washed with water (3x), dried over
MgS04 and evaporated to dryness to afford the crude product, which was further
purified by column chromatography.
.
GENERALPROCEDURE AQ
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Fourth Transesterification Technigue
The ester to be transesterified was dissolved in a large excess of the
alcohol and 0.3 equivalents of titanium(IV) isopropoxide (Aldrich) was added.
The reaction was followed by tlc until complete and then the volatiles were
removed at reduced pressure. The resulting crude material was then
chromatographed to obtain the desired product.
GENERALPROCEDURE AR
Synthesis of N BOC Anilines
To a solution of the aniline in THF was added dropwise 1 equivalent of
di-tert-butyl dicarbonate (Aldrich) in THF and then I.5 equivalents of lON
aqueous sodium hydroxide at 0°C. After stirring at room temperature for
16
hours, or heating at 80°C for 3 hours, if needed, the reaction mixture
was
diluted with ether and washed with NaHC03, brine, dried over sodium sulfate
and potassium carbonate, concentrated at reduced pressure and chromatographed
to afford the N BOC aniline.
GENERAL PROCEDURE AS
Oxime Ester Formation
The trichlorophenyl ester (1 eq.) was stirred in DMF or THF. The
oxime (I.2 eq.) was added and the mixture was stirred at ambient temperature
for I to 4 hours. The resulting mixture was concentrated under reduced
pressure
and the residue was purified by silica gel chromatography and/or
crystallization.
Example AA
Synthesis of D,L-alanine iso-butyl ester hydrochloride
A mixture of 35.64 g (0.4 mol) of D,L-alanine (Aldrich), 44 mL (0.6
mol) of thionyl chloride (Aldrich) and 200 mL of iso-butanol was refluxed for
1.5 hours. The volatiles were removed at reduced pressure at 90°C under
reduced pressure to give the title compound as an oil, which was used without
further purification.
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NMR data was as follows:
~H-nmr (CDCI,): b = 8.72 (br s, 3H), 4.27 (q, 3 = 7.4 Hz, IH), 3.95 (m,
2H), 1.96 (s; I H), 1.73 (d, J = 7.2 Hz, 31-1), 0.92 (d, J = 6.7 Hz, 6H).
~'C-nmr {CDC13): 8 = 170.0, 72.2, 49.2, 27.5, 18.9, 16. Y.
Example AB
Synthesis of N-(3,4-dichlorophenyl)alanine
Using the procedure set forth in U.S. Patent No. 3,598,859, the
disclosure of which is incorporated herein by reference in its entirety, N-
{3,4-
dichlorophenyl)alanine was prepared. Specifically, to a solution of 3,4-
dichloroaniline ( I equivalent) (Aldrich) in isopropanol (about 500 mL per
mole
of 3,4-dichloroaniline) is added water (about 0.06 mL per mL of isopropanol)
and 2-chloropropionic acid (2 equivalents) (Aldrich). This mixture is warmed
to
40°C and sodium bicarbonate (0.25 equivalents) is added in successive
portions
before heating under reflux for 4-5 days. After cooling, the reaction mixture
is
poured into water and the unreacted 3,4-dichloroaniline is removed by
filtration.
The filtrate is acidified to pH 3-4 with concentrated hydrochloric acid and
the
resultant precipitate is filtered, washed and dried to yield the title
compound,
m.p. = 148-149°C.
Alternatively, following General Procedure AF above and using N-(3,4-
dichlorophenyl)alanine ethyl ester (from Example A 1 below), the title
compound
was prepared.
Example AC
Synthesis of N-(3,5-difluorophenyl)alanine
Using the procedure set forth in U.S. Patent No. 3,598,859, N-(3,5-
difluorophenyl)alanine was prepared using 3,5-difluoroaniline (Aldrich) and 2-
chloropropionic acid (Aldrich).
Example AD
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Synthesis of Iso-butyl 2-bromopropionate
To a mixture of iso-butanol and 1.0 equivalent of pyridine in dry diethyl
ether was added dropwise 1.3 equivalents of 2-bromopropionyl bromide at
0°C.
After stirring at room temperature for 16 hours, the reaction was diluted with
diethyl ether, washed with 1N HC1, water, aqueous NaHC03, brine and dried
over magnesium sulfate or sodium sulfate. Removal of the solvents at reduced
pressure gave the title compound as a clear oil.
Example AE
Synthesis of N-(2-naphthyl)alanine 2,4,5-trichlorophenyl ester
N-(2-Naphthyl)alanine methyl ester (5.0 g, 20.6 mmol) (from Example
A44 below) was dissolved in dioxane (100 mL). NaOH (30 mL, 1N) was added
and the resulting solution was stirred for 1 hour. The reaction mixture was
concentrated under reduced pressure. The resulting solid was dissolved in
water
and the aqueous mixture was washed with ether. The aqueous portion was
adjusted to pH 3 with IN HCl and extracted with ethyl acetate. The organic
extracts were dried over magnesium sulfate or sodium sulfate and concentrated
under reduced pressure to yield a white solid (4.35 g, 98%).
The resulting solid (4.35 g, 20 mmol) was dissolved in dichloromethane
(300 mL). 2,4,5-Trichlorophenol (4.9 g, 25 mmol) (Aldrich) was added
followed by dicyclohexylcarbodiimide (25 mL, 1 M in dichloromethane)
(Aldrich). After stirring for 18 hours, the mixture was filtered and
concentrated
to provide an oil which was purified by chromatography on silica gel using
chloroform as the eluant (Rf = 0.6). The title compound was obtained as a
thick
oil which slowly crystallized.
Example A 1
Synthesis of N-(3,4-dichlorophenyl)alanine ethyl ester
Following General Procedure AA above and using 3,4-dichloroaniline
(Aldrich) and ethyl pyruvate (Aldrich), the title compound was prepared as an
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oil. The reaction was monitored by tlc on silica gel (Rf = 0.4 in 25%
EtOAc/hexanes) and purification was by preparative plate chromatography
(silica
gel using 25% EtOAc/hexanes as the eluant).
NMR data was as follows:
S 'H-nmr (CDCI,): & = 7.2 (d, 1 H); 6.7 (d, 1 H,); 6.4 (dd, 1 H); 4.30 {bs,
1 H); 4.2 {q, 2H); 4.1 (q, 1 H); 1.5 (d, 3H); 1.3 (t, 3H).
'''C-nmr (CDC13): 8 = 175; 146.7; 133; 131; 121; 114.9; 112.6; 72.0;
52.4; 28.3; 19.5.
C"H,3C1,N0, (MW = 262.14).
Example A2
Synthesis of N-(3-trifluoromethyl-4-chlorophenyl)alanine ethyl ester
Following General Procedure AA above and using 4-chloro-3-
(trifluoromethyl)aniline (Aldrich) and ethyl pyruvate (Aldrich), the title
compound was prepared.
Analysis: Calc.: C, 48.74; H, 4.43; N, 4.74. Found: C, 48.48; H, 4.54;
N, 4.94.
C,~H,3F3CIN0, (MW = 295.69); mass spectroscopy (MH') 295.
Example A3
Synthesis of N-(3,5-dichlorophenyl)alanine ethyl ester
Following General Procedure AA above and using 3,5-dichloroaniline
(Aldrich) and ethyl pyruvate (Aldrich), the title compound was prepared.
Analysis: Calc.: C, 50.40; H, 5.00; N, 5.34. Found: C, 50.50; H, 5.06;
N, 5.25.
C"H,3Cl,N0z (MW = 262.14); mass spectroscopy (MH+) NA.
Example A4
Synthesis of N-(3,4-difluorophenyl)alanine ethyl ester
Following General Procedure AA above and using 3,4-difluoroaniline
(Aldrich) and ethyl pyruvate (Aldrich), the title compound was prepared. The
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reaction was monitored by tlc on silica gel (Rf = 0.4 in 25% EtOAe/hexanes)
and purification was by preparative plate chromatography (silica gel using 25%
EtOAc/hexailes as the eluant).
NMR data was as follows: -
'H-nmr {CDC1,): 8 = 7.4 (m, 1 H), 6.8 (d, 1 H), 6.5 (m, 1 H), 4.30 (bs,
1H), 4.2 (q, 2H), 4.1 (q, 1H), 1.5 (d, 3H), 1.3 (t, 3H).
''C-nmr (CDC13): b = 175, 146.7, 135, 132, 125, 116, 113, 72, 52, 28,
19.
C"H,3F~N0, {MW = 229.23); mass spectroscopy (MH') 230.
Example AS
Synthesis of N-(3,4-dichlorophenyl)alanine benzyl ester
Following General Procedure AA above and using 3,4-dichloroaniline
(Aldrich) and benzyl pyruvate (prepared by following General Procedure AO
above using benzyl alcohol in place of iso-butanol), the title compound was
prepared as an oil. The reaction was monitored by tle on silica gel (Rf = 0.4
in
25% EtOAc/hexanes) and purification was by preparative plate chromatography
(silica gel using 25% EtOAc/hexanes as the eluant).
NMR data was as follows:
'H-nmr (CDCI,): 8 = 7.18 (d, 1 H); 7.0 (nl. SH); 6.6 (d, 1 H,); 6.4 (dd,
I I-I); 5.1 (s, 2H}; 4.30 (bs, 1 H); 4.08 (q, 1 H); 1.94 (m, 1 H); 1.47 (d,
3H); 0.91
(d, 6H).
''C-nmr (CDC13): 8 = 174.5; 146.7; 133.5; 131.3; 121.3; 120.1; 114.9;
113.6; 72.0; 60.1; 52.4; 28.3; 19.5; 19.3.
C,6H,SC1~N0, (MW = 324.31); mass spectroscopy (MH') 325.
Example A6
Synthesis of N-(3,4-dichlorophenyl)alanine iso-butyl ester
Following General Procedure AA above and using 3,4-dichloroaniline
(Aldrich) and iso-butyl pyruvate (prepared by following General Procedure AO
above), the title compound was prepared as an oiI. The reaction was monitored
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by tlc on silica gel (Rf = 0.55 in 25% EtOAc/hexanes) and purification was by
preparative plate chromatography (silica gel using 25% EtOAc/hexanes as the
eluant).
NMR data was as follows:
'H-nmr (CDCI,): S = 7.18 {d, 1 H, J=8.7 Hz), 6.66 (d, 1 H, J=2.7 Hz),
6.43 (dd, 1 H, J = 8.7 Hz, J = 2.7 Hz), 4.30 (bs, 1 H), 4.08 (q, 1 H, J = 6.9
Hz),
1.94 (sept, 1H, J = 6.7 Hz), 1.47 (d, 3H, J = 6.9 Hz), 0.91 (d, 6H,
J = 6.6 Hz).
'3C-nmr (CDCl3) 8 = 174.5, 14b.7, 133.5, 131.3, 121.3, 114.9, 113.6,
72.0, 52.4, 28.3, 19.5, 19.3.
C,3H"CI~NO, (MW = 290.19); mass spectroscopy (MH-) 290.
Example A7
Synthesis of N-(3,4-dichlorophenyl)alanine iso-propyl ester
Following General Procedure AA above and using 3,4-dichioroaniline
(Aldrich) and isopropyl pyruvate (prepared by following General Procedure AO
above using isopropanol in place of iso-butanol), the title compound was
prepared as an oil. The reaction was monitored by tlc on silica gel (Rf = 0.4
in
25% EtOAc/hexanes) and purification was by preparative plate chromatography
(silica gel using 25% EtOAc/hexanes as the eiuant).
NMR data was as follows:
'H-nmr (CDCI,): F~ = 7.18 (d, 1 H,>; 6.66 (d, 1 H,); 6.43 {dd, 1 H); 4.30
(bs, 1 H); 4.08 (m, 1 H); 1.94 (m, 1 H); 1.47 (d, 3H); 0.91 (d, 6H).
'3C-nmr (CDCl3): 8 = 174.5; 146.7; 133.5; 131.3; 121.3; 114.9; 113.6;
72.0; 52.4; 19.5.
C,~H,SC1,N0~ (MW = 276.16); mass spectroscopy (MH~) 277.
Example A8
Synthesis of N-(3,4-dichlorophenyl)alanine n-butyl ester
Following General Procedure AA above and using 3,4-dichloroaniline
(Aldrich) and n-butyl pyruvate (prepared by following General Procedure AO
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above using n-butanol in place of iso-butanol), the title compound was
prepared.
The reaction was monitored by tlc on silica gel (Rf = 0.7 in 25%
EtOAc/hexaries) and purification was by preparative plate chromatography
(silica
gel using 25% EtOAc/hexanes as the eluant). -
NMR data was as follows:
'H-nmr (CDCI,): S = 7.18 (d, 1H); 6.66 (d, 1H,); 6.43 (dd, II-I); 4.30
(bs, 1 H); 4.2 (m, 2H}; 4.08 (q, 1 H); 1.94 (m, 1 H); 1.47 (m, 4H); 0.91 (t,
3H).
''C-nmr (CDCl3): 8 = 174.5; 146.7; 133.5; 131.3; 121.3; 114.9; 113.6;
72.0; 52.4; 28.3; 20.2; 19.5.
C,3H"CI,NO~ (MW = 290.19); mass spectroscopy (MH+) 291.
Example A9
Synthesis of N-(3,4-dichlorophenyl)alanine methyl ester (R,S isomers)
Following General Procedure AA above and using 3,4-dichloroaniline
(Aldrich) and methyl pyruvate {Aldrich), the title compound was prepared as an
oil. The reaction was monitored by tlc on silica gel (Rf = 0.5~ in 25%
EtOAc/hexanes) and purification was by flash chromatography (silica gel using
25% EtOAc/hexanes as the eluant).
NMR data was as follows:
'H-nmr (CDCI,): 8 = 7.19 (d, J = 8.73 Hz, 1H), 6.66 (d, J = 2.75 Hz,
1 H), 6.43 (dd, J = 8.73 Hz, 2.80 Hz, 1 H), .4.25 (bd, J = 8.25 Hz, 1 H), 4.08
(m,
IH), 3.76 (s, 3H), 1.47 (d, J = 6.90 Hz).
'3C-nmr (CDCl3) s = 174.35, 145.96, 132.87, 130.70, 120.76, 114.38,
112.90. 52.43, 51.70, 18.67.
C,oH"CIzNO, (MW = 248.1 I }; mass spectroscopy (MH+) 247.
Example A10
Synthesis of N-(3,4-dichlorophenyl)alanine cyclopentyl ester
Following transesterification General Procedure AB above and using N-
(3,4-dichlorophenyl)alanine methyl ester (from Example A9 above} and
...a.-. . ,
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cyclopentanol {Aldrich), the title compound was prepared as an oil. -The
reaction was monitored by silica gel tlc (Rf = 0.66 in 25% EtOAc/hexanes).
PurificatioW vas by preparative plate chromatography (silica gel using 25%
EtOAc/hexanes as the eluant).
NMR data was as follows:
'H-nmr (CDCI,): 8 = 7.19 (d, 1H, J = 8.7 Hz), 6.66 (d, 1H, J = 2.7 Hz),
6.43 (dd, 1 H) J = 8.7 Hz, J = 2.7 Hz), 5.2'? (m, 1 H), 4.27 (d, 1 H,
J = 8.I Hz), 4.02 (quint, 1H, J = 7.5 Hz), 1.74 (m, 8H), I.43 (d, 3H,
J=6.9Hz).
"C-nmr (CDC13): 8 = 174.3, 146.7, 133.4, 131.2, 121.2, 114.9, 113.7,
78.9, 52.5, 33.2, 24.2, 24.1, 19.1.
C,4H,.,CI,NO~ (MW = 302.20); mass spectroscopy (MHT) 301.
Example A11
Synthesis of N-(3,4-dichlorophenyl)alanine n-propyl ester
Following General Procedure AA above and using 3,4-dichloroaniline
(Aldrich) and n-propyl pyruvate (prepared by following General Procedure AO
above using n-propanol in place of iso-butanol), the title compound was
prepared as an oil. The reaction was monitored by tlc on silica gel (Rf = 0.5
in
25% EtOAc/hexanes) and purification was by preparative plate chromatography
(silica gel using 25% EtOAc/hexanes as the eluant).
NMR data was as follows:
'H-nmr (CDC1,): 8 = 7.2 (d, 1 H); 6.6 (d, 1 H); 6.4 (dd, 1 H); 4.30 (bs,
1H); 4.2 (q, 21-1); 4.08 (q, 1H); 1.94 (m, 2H); 1.5 (d, 3H); 0.95 (t, 3H).
'3C-nmr (CDC13): 8 = 178; 144.7; 130.2; 120.62; 115.11; 71.82; 52.90.
C,,H,5C1,N0, (MW = 276.16); mass spectroscopy (MHY) 277.
Example A 12
Synthesis of N-(3,4-dichlorophenyl)alanine allyl ester
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Following transesterification General Procedure AB above and using N-
(3,4-dichlorophenyl)alanine methyl ester (from Example A9 above) and allyl
alcohol (Aldrich), the title compound was prepared as an oil. The reaction was
monitored by silica gel tlc (Rf = 0.62 in 25% EtOAc/hexanes).- Purification
was
by preparative plate chromatography (silica gel using 2~% EtOAclhexanes as the
eluant).
NMR data was as follows:
' H-nmr (CDCl3}: 8 = 7.19 (d, 1 H, J = 8.7 Hz), 6.67 (d, 1 H, J = 2.8 Hz),
6.44 (dd, 1 H, J = 8.7 Hz, J=2.8 Hz), 5.90 (m, 1 H), 5.30 (m, 2H), 4.64 (m,
2H},
4.26 (m, 1 H, 4.10 (m, I H), I .48 (d, 3H, J = 6.9 Hz).
'3C-nmr (CDCI3): b = 174.1, 146.6, 133.5, 132.1, 131.3, 121.4) 119.6,
i 15.0, 113.6, 66.5, 52.4, 19.3.
C,,H,3C1~N0, (MW = 274.15); mass spectroscopy (MH~) 273.
Example A 13
Synthesis of N-(3,4-dichlorophenyi)alanine 4-methylpentyl ester
Following transesterification General Procedure AB above and using N-
(3,4-dichlorophenyl)alanine methyl ester {from Example A9 above) and 4-
methylpentanol (Aldrich), the title compound was prepared as an oil. The
reaction was monitored by silica gel tlc (Rf = 0.70 in 25% EtOAc/hexanes).
Purification was by preparative plate chromatography (silica gel using 25%
EtOAclhexanes as the eluant).
NMR data was as follows:
'H-nmr (CDCI,): 8 = 7.18 (d, 1H, J = 8.7 Hz), 6.66 (d, 1H, J = 2.7 Hz),
6.43 (dd, 1 H, J = 8.7 Hz, J = 2.7 Hz}, 4.28 (m, 1 H), 4.10 {m, 31-I), 1.55
(m,
6H), 1.19 {m, 2H), 0.87 (d, 3H, J = 6.6 Hz).
'3C-nmr (CDC13): 8 = 174.6, 146.7, I33.4, 131.3, 121.3, 115.0, 113.6,
66.4, 52.4, 35.4, 28.2, 27.0, 23.0, 19.3.
C,SH~~C1~N0, (MW = 318.25); mass spectroscopy (MH+) 317.
Example A 14
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Synthesis of N-(3,4-dichlorophenyl)alanine 2,2-dimethyl-1,3-dioxolane-4-
- methyl ester
Following transesterification General Procedure AB above and using N-
(3,4-dichlorophenyl)alanine methyl ester (from Example A9 above) and 2,2-
dimethyl-1,3-dioxolane-4-methanol (solketal) (Aldrich), the title compound was
prepared as a mixture of diastereomers. The reaction was monitored by silica
gel tlc (Rf = 0.32 in 25% EtOAc/hexanes). Purification was by preparative
plate chromatography (silica gel using 25% EtOAc/hexanes as the eluant).
NMR data was as follows:
'H-nmr (CDC1,): b = 7.19 (d, 1 H, J = 8.7 Hz), 6.66 (d, 1 H, 2.7 Hz),
6.43 (dd, 1 H, J = 8.7 Hz, J = 2.7 Hz}, 4.22 (m, 61-I}, 3.70 (m, 1 H), I .43
(m,
9H).
''C-nmr (CDC13): 8 = 174.34, 174.32, 146.5, 133.5, 131.3, 121.5, 115.0,
113.6, 110.52, 110.51, 73.97, 73.89, 66.6, 66.01, 65.95, 52.42, 52.37, 27.3,
25.8, 19.3.
C,5H,9C1,N04 {MW = 348.23); mass spectroscopy {MH+) 347.
Example A 15
Synthesis of N-(3,4-dichlorophenyl)alanine cyclohexylmethyl ester
Following transesterification General Procedure AB above and using N-
(3,4-dichlorophenyl)aianine methyl ester (from Example A9 above) and
cyclohexylmethanol (Aldrich), the title compound was prepared.
NMR data was as follows:
'H-nmr (CDCI,): 8 = 7.19 (d, 1H), 6.68 (d, 1H), 6.45 (dd, 1H), 4.26
(bd, 1 H), 4.10 (m, I H), 3.95 (d, 2H}, 1.70- I .5 5 (m, 6H}, 1.50 (d, 3 H),
1.3 ~-0.85
(m, 5H).
''C-nmr (CDC13): 8 = 174.58, 146.72, 133.48, 131.27, 121.34, 114.98,
113.72, 71.06, 52.52, 37.68, 30.10, 26.83, 26.17, 19.32.
C,SH~,CI~NO~ (MW = 318.25); mass spectroscopy (MH+) 317.
Example A16
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Synthesis of N-(3,4-dichlorophenyl)alanine tert-butyloxycarbonylmethyt
ester
Following General Procedure AE above and using N-(3,4-
dichlorophenyl)alanine (from Example AB above) and tert-butyl bromoacetate
(Aldrich), the title compound was prepared as a solid. The reaction was
monitored by silica gel tlc (Rf = 0.57 in 25% EtOAc/hexanes). Purification was
by recrystallization from ethanol.
NMR data was as follows:
'H-nmr (CDCI,): 8 = 7.19 (d, 1H), 6.68 (d, 1H), 6.45 (dd, 1H), 4.55 (m,
2H), 4.20 (m, 2H), 1.55 (d, 3H), 1.45 (s, 9H).
''C-nmr (CDC13): 8 = 173.9, 166.9, 146.5, 133.5, 131:3, 115.1, 113.6,
83.4, 62.2, 52.2, 28.6, 19.3.
C,SH,9C1,N0,, (MW = 348.23); mass spectroscopy (MH+) 347.
Example A 17
Synthesis of N-(3,4-dichlorophenyl)leucine iso-butyl ester
Following General Procedure AA above and using 3,4-dichloroaniline
(Aldrich) and iso-butyl 4-methyl-2-oxopentanoate (prepared by following
General Procedure AO above using 4-methyl-2-oxovaleric acid (Fluka) and
iso-butanol), the title compound was prepared as an oil. The reaction was
monitored by tlc on silica gel (Rf = 0.6 in 25% EtOAc/hexanes) and
purification
was by preparative plate chromatography (silica gel using 25% EtOAc/hexanes
as the eiuant).
NMR data was as follows:
'H-nmr (CDCI,): 8 = 7.2 (d, 1H}; 6.5 (d, IH); 6.4 (dd, IH}; 4.30 (bs,
1 H); 4.08 (q, 1 H); 3.8(m, 2H); 1.8 (m, 3 H); 0.91 (m, 12H).
'3C-nmr (CDC13): 8 = 174.5; 146.7; 133.5; 131.3; 121.3; I 14.9; 113.6;
72.0; 52; 28.3; 20.1; 19.5.
C,6H23CI,N0, (MW = 332.27); mass spectroscopy (MH+) 333.
Example A 18
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Synthesis of 2-[N-(3,4-dichlorophenyl)amino]pentanoic acid iso-butyl ester
Following General Procedure AA above and using 3,4-dichloroaniline
' (Aldrich) and iso-butyl 2-oxopentanoate (prepared by following General
Procedure AO above using 2-oxovaleric acid (Fluka) and iso-butanol), the title
compound was prepared as an oil. The reaction was monitored by tlc on silica
gel (Rf = 0.5 in 25% EtOAc/hexanes) and purification was by preparative plate
chromatography (silica gel using 25% EtOAc/hexanes as the eluant).
NMR data was as follows:
'H-nmr (CDCI,): 8 = 7.2 (d, 1 H); 6.6 (d, I H); 6.4 (dd, 1 H); 4.3 (d, 1 H);
3.8 (m, 3H); 1.9 (m, 6H); 1.0 (t, 3H}, 0.9 (m, 6H}.
''C-nmr (CDC13): F~ = 178; 144.7; 130.2; 120.62; 115.11; 71.82; 52.90;
28.30; 19.53.
C,SH,,CI,NO, (MW = 318.3); mass spectroscopy (MH*) 319.
Example A19
Synthesis of N-(4-cyanophenyl)alanine iso-butyl ester
Following General Procedure AP above and using 4-fluorobenzonitrile
(Aldrich) and D,L-alanine iso-butyl ester hydrochloride {from Example AA
above), the title compound was prepared as an oil. The product was recovered
by column chromatography on silica gel using 1:5 EtOAc/hexanes as the eluant.
NMR data was as follows:
'H-nmr (CDCI,): 8 = 7.44 (d, J = 8.8 Hz, 2H), 6.57 (d, J = 8.8 Hz, 2H},
4.74 (d, J = 8.1 Hz, I H}, 4.18 (t, J = 7.4 Hz, I H), 3.95 (m, 2H), 1.94 (m, 1
H),
1.51 (d, J = 6.9 Hz, 3H), 0.91 (d, J = 6.7 Hz, 6H).
'3C-nmr (CDC13): 8 = 173.4, 149.7, 133.8, 120.1, 112.7, 99.8, 71.6, 51.2,
27.7, 18.9, 18.6.
C,4H,8N,0z MW = 246.31; mass spectroscopy (MH+) 247.
Example A20
Synthesis of N-(3-chloro-4-cyanophenyl)alanine iso-butyl ester
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Following General Procedure AP above and using 2-chloro-4-
fluorobenzonitrile (Aldrich) and D,L-alanine iso-butyl ester hydrochloride
(from
Example AA above), the title compound was prepared. The product was
recovered by column chromatography on silica gel using 1:5 EtOAc/hexanes as
the eluant.
NMR data was as follows:
' H-nmr (CDCI,): ~ = 7.40 (d, J = 8.5 Hz, 1 H), b.62 (d, J = 2.3 Hz, 1 H),
6.48 (dd, J = 2.4, 8. 6 Hz, 1 H ), 4.90 (d, J = 7. 6 Hz, 1 H j, 4.16 (quintet,
J = 7.1
Hz, 1 H), 3.96 (dd, J = 2.2, 6.7 Hz, 2H), 1.97 (m, 1 H), 1.51 (d, J = 7.0 Hz,
3H),
0.93 (d, J = 6.7 Hz, 6H).
'3C-nmr (CDC13): S = 173.0, 150.4, 138.3, 134.9, 117.3, 112.8, 111.3,
I 00.6, 71.7, 51.1, 27.7, 18.9, 18.4.
C,4H"N,O,CI MW = 280.76; mass spectroscopy (MH+) 281.
Example A21
Synthesis of N-(3,4-dichloro)alanine iso-butyl ester (S isomer)
Following General Procedure AM above and using 3,4-dichloroaniline
(Aldrich) and iso-butyl R-(+)-lactate (Aldrich), the title compound was
prepared
as an oil. The reaction was monitored by silica gel tlc (Rf = 0.55 in 25%
EtOAc/hexanes). Purification was column chromatography.
NMR data was as follows:
H-nmr (CDCI,): 8 = 7.19 (d, J = 8.73, 1 H), 6.67 (d, J = 2.7~, 1 H), 6.45
(dd, J = 8.73, J = 2.75, 1 H), 4.28 (bd, J = 8.36, 1 H), 4.09 (quint, 1 H),
3.94 {d, J
= 6.66, 2H), 1.95 (hept, J = 6.71, 1 H), 1.49 (d, J = 6.90, 3 H), 0.92 (d, J =
6.04,
6H).
'3C-nmr {CDC13): 8 = 174.57, 146.67, 133.47, 131.28, 121.29, 114.93,
113.63, 71.01, 52.43, 28.30, 19.55, 19.33.
C,3H"C1,N02 (MW = 290.19); mass spectroscopy (MH+) 290.
Example A22
.. ... ~_.......,..""~ ...._....... . . . r
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Synthesis of N-(3,4-dichloro)alanine tetrahydrofuran-3-yl-methyl ester
Following transesterification General Procedure AB above and using N-
(3,4-dichlorophenyl)alanine methyl ester (from Example A9 above) and
tetrahydro-3-furanmethanol (Aldrich), the title compound was prepared as an
oil.
The reaction was monitored by silica gel tlc (Rf = 0.33 in 25% EtOAc/hexanes).
Purification was by preparative plate chromatography (silica gel using 25%
EtOAc/hexanes as the eluant).
NMR data was as follows:
'H-nmr (CDCI,): 8 = 7.18 (d, 1H, J = 8.7 Hz), 6.65 (d, 1H, J = 2.7 Hz),
6.42 (dd, 1H, J = 8.7 Hz, J = 2.7 Hz), 4.30 (m, 1I-I), 4.09 (m, 3H), 3.78 (m,
3H), 3.53 (m, 1H}, 2.56 (m, 1H), 1.94 (m, 1H), 1.58 (m, 1H), 1.46 (d, 3H, J =
6.9 Hz).
'3C-nmr (CDC13}: 8 = 174.5, 146.6, 133.5, 131.3, 121.4, 114.9, 113.6,
70.86, 70.83, 68.2, 67.31, 67.29, 52.4, 38.7, 29.36, 29.33, 19.2.
C,4H"C1,N03 (MW = 318.20); mass spectroscopy (MH~) 318.
Example A23
Synthesis of N-{3,5-dichlorophenyl)alanine n-propyl ester
Following General Procedure AA above and using 3,5-dichloroaniline
(Aldrich) and n-propyl pyruvate (which can be prepared by following General
Procedure AO above using n-propanol in place of iso-butanol), the title
compound could be prepared.
Example A24
Synthesis of 2-(N-(3,4-dichlorophenyl)amino] butanoic acid iso-butyl ester
Following General Procedure AA above and using 3,4-dichioroaniline
(Aldrich) and iso-butyl 2-oxobutanoate (prepared by following General
Procedure AO above using 2-oxobutyric acid (Aldrich) and iso-butanol), the
title
compound was prepared as an oil. The reaction was monitored by tlc on silica
gel (Rf = 0.3 in 25% EtOAc/hexanes) and purification was by preparative plate
chromatography (silica gel using 25% EtOAc/hexanes as the eluant).
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NMR data was as follows:
'H-nmr (CDCI,): S = 7.2 (d, 1 H); 6.6 (d, 1 H); 6.4 (dd, 1 H); 4.3 (d, 1 H);
3.8 {m, 3H); 1.9 (m, 3H); 1.0 (t, 3H); 0.9{m, 6H).
''C-nmr (CDCI3): 8 = 178; 144.7; 130.2; 120.62; 115.11-; 71.82; 52.90;
28.30; 20.5; 19.53.
C,4H,9Cl,N0z (MW = 304.22); mass spectroscopy (MH+) 305.
Example A25
Synthesis of N-(4-chlorophenyl)alanine iso-butyl ester
Following General Procedure AA above and using 4-chloroaniline
(Aldrich) and iso-butyl pyruvate (prepared by following General Procedure AO
above), the title compound was prepared as an oil. The reaction was monitored
by tlc on silica gel (Rf = 0.6 in 25% EtOAc/hexanes) and purification was by
preparative plate chromatography (silica gel using 25% EtOAclhexanes as the
eluant).
NMR data was as follows:
'H-nmr (CDCI,): 8 = 7.18 (d, 2H), 6.66 (d, 2H), 4.30 (bs, 1H), 4.08 (q,
1 H), 1.94 (sept, 1 H), 1.47 (d, 31-1), 0.91 (d, 6H).
'3C-nmr (CDC13): b =174.5, 146.7, 133.5, 131.3, 121.3, 114.9, 113.6,
72.0, 52.4, 28.3, 19.5, 19.3.
C,3H,gCINO~ (MW = 255.75); mass spectroscopy (MH+) 256.
Example A26
Synthesis of N-(3,5-dichlorophenyl)alanine iso-butyl ester
Following General Procedure AA above and using 3,5-dichloroaniline
(Aldrich) and iso-butyl pyruvate (prepared by following General Procedure AO
above), the title compound was prepared as an oil. The reaction was monitored
by tlc on silica gel (Rf = 0.4 in 25% EtOAc/hexanes) and purification was by
preparative plate chromatography (silica gel using 25% EtOAc/hexanes as the
eluant). '
NMR data was as follows:
_. ....TM. ~
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'H-nmr (CDCI,): b = 7.18 (d, 2H), 6.66 (m, 1H), 4.30 (bs, 1H), 4.08 (q,
1 H), 1.94 (m, 1 H), 1.47 {d, 3H), 0.91 (d, 6H).
' '''C-ninr (CDC13): b = 175; 146.7; 133; 131; 121; 114.9; 112.6; 72.0;
52.4; 28.3; 19.5.
C,3H"C1,N0, (MW = 290.2); mass spectroscopy (MH') 291.
Example A27
Synthesis of N-(4-ethylphenyl)alanine methyl ester
A solution of 0.68 g (5 mmol) of 4'-aminoacetophenone (Aldrich), 0.60
mL of 90% methyl pyruvate (Aldrich) and 0.05 g (0.25 mmol) of p-
toluenesulfonic acid in ethanol was hydrogenated in the presence of a
catalytic
amount of 10% Pd/C at from 30 to 15 psi of hydrogen for 16 hours. The
catalyst was removed by filtering the reaction mixture through Celite and the
solvent was evaporated to provide the crude product. The product was purified
by column chromatography (silica gel using 1:9 EtOAc/hexanes as the eluant) to
provide the title compound.
NMR data was as follows:
'H-nrnr (CDC1,): 8 = 1.19 (t, J = 7.6 Hz, 3H), 1.47 (d, J = 6.8 Hz, 3H),
2.54 (q, J = 7.6 Hz, 2H), 3.74 (s, 3H), 4.04 (bs, 1 H), 4.13 (m, 1 H), 6.57
(d, J =
8.5 Hz, 2H), 7.03 (d, J = 8.4 Hz, 2H).
'3C-nmr (CDCl3): 8 = 15.8, 18.0, 27.9, 52.17, 52.19, 113.5, 128.6, 134.1,
144.4, 175.3.
C,,H"NO, MW = 207.27; mass spectroscopy (MH+) 208.
Example A28
Synthesis of N-(4-(1-ethoxy)ethylphenyl)alanine methyl ester
Following the procedure for Example A27 above, the title compound was
isolated as another reaction product by column chromatography (silica gel
using
1:9 EtOAc/hexanes as the eluant).
NMR data was as follows:
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'H-nmr (CDC1,}: 8 = 1.15 (t, J = 7.0 Hz, 3H), 1.40 (d, J = 6.5 Hz, 3H),
1.47 (d, J = 6.1 Hz, 3H), 3.31 (q, J = 5.1 Hz, 2H), 3.74 (s, 3H), 4.14 (m,
2H),
4.29 (q, J = 6.4 Hz, 1H}, 6.57 (d, .1 = 8.5 Hz, 2H), 7.I2 (d, J = 8.4 Hz, 2H).
''C-nmr (CDC13): 8 = 15.4, 19.0, 23.9, 51.9, 52.2, 63.4, 77.3, 113.1,
127.3, 133.6, 145.8, 175.1.
C,4H,'N03 MW = 251.33; mass spectroscopy (MH+) 251.
Example A29
Synthesis of N-(3,4-dichloro)alanine 2,2-dimethylpropyl ester
(R,S isomers)
Following transesterification General Procedure AQ above and using N-
(3,4-dichlorophenyl)alanine methyl ester (from Example A9 above) and
neopentyl alcohol (Aldrich), the title compound was prepared. The reaction was
monitored by silica gel tlc (Rf = 0.72 in 25% EtOAc/hexanes). Purification was
by flash chromatography (silica gel using 25% EtOAc/hexanes as the eluant).
NMR data was as follows:
'H-nmr (CDCI,): b = 7.19 (d, 1 H, J = 8.7 Hz), 6.68 (d, 1 H, J = 2.7 Hz),
6.45 (dd, 1 H, J = 8.7 Hz, J = 2.7 Hz}, 4.29 (m, 1 H), 4.11 (m, 1 H), 3.85 (m,
2H), 1.49 (d, 3H, .T = 6.9 Hz}, 0.93 (s, 9H).
''C-nmr (CDC13): 8 = 174.6, 146.7, 133.5, 131.3, 121.3, 114.9, 113.7,
75.2, 52.4, 32.0, 26.9, 19.4.
C',,H'9C1,N0, (MW = 304.22); mass spectroscopy (MH') 303.
Example A30
Synthesis of N-(3,4-dichlorophenyl)glycine iso-butyl ester
3,4-Dichloroaniline (Aldrich) was treated with di-tert-butyl dicarbonate
(Aldrich) using conventional procedures to produce the N-BOC aniline. The N
BOC aniline was treated with sodium hydride in THF and then with iso-butyl 2
bromoacetate (from Example AD above) to produce the N-BOC N-(3,4
dichlorophenyl)glycine iso-butyl ester. The BOC group was then removed using
General Procedure AN above to afford the title compound. The reaction was
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monitored by tlc on silica gel (Rf = 0.78 in 50% EtOAc/hexanes) and
purification was by preparative plate chromatography (silica gel using 50%
" EtOAc/hexanes as the eluant).
NMR data was as follows: -
'H-nmr (CDCI,): 8 = 7.19 (dd, J=4.1, 4.7, 3.4, 1H); 6.65 {d, J=2.7, 1H);
6.44 (dd, J=2.7, 4.5, 4.2, 1 H): 4.4 (m, 1 H): 3.97 (dd, J=3.6, 3.0, 2.3, 2H);
3.87
(s, 2H); 1.9 (m, 1 H); 0.93 (d, J=6.7, 6H).
''C-nmr (CDC13): 8 = 171.2, 147.0, 133.5, 131.3, 121.2, 114.5, 113.3,
72.2, 46.0, 28.2, 19.6.
IO C,,H,SCI,NO~ (MW = 276); mass spectroscopy (MH') 277.
Example A31
Synthesis of N-(3,4-dichlorophenyl)alanine 2-ethylbutyl ester
Following General Procedure AA above and using 3,4-dichloroaniline
(Aldrich) and 2-ethylbutyl pyruvate (prepared by following General Procedure
AO above using 2-ethylbutanol (Aldrich) in place of iso-butanol), the title
compound was prepared as an oil. The reaction was monitored by tlc on silica
gel (Rf = 0.6 in 25% EtOAc/hexanes) and purification was by preparative plate
chromatography (silica gel using 25% EtCIAc/hexanes as the eluant).
NMR data was as follows:
' H-nmr {CDCI,): b = 7.2 (d, 1 H); 6.6 (d, 1 H); 6.4 (dd, 1 H); 4.2 (t, 2H);
4.1 (q, 1 H); 1.5 (d, 3H); 1.4 (m, 4H); 1.0 (m, 6H).
''C-nmr (CDC13): 8 = 178; 144.7; 130.2; 120.62; 115.11; 70.7; 51.90;
26.3; 19.53, 18.5.
C,SH~,CI,NOz (MW = 318.25); mass spectroscopy (MH') 319.
Example A32
Synthesis of N-(3-chloro-4-iodophenyl)alanine iso-butyl ester
Following General Procedure AR above and using 3-chloro-4-iodoaniline
(Aldrich), N BOC-3-chloro-4-iodoaniline was prepared. To a stirred slurry of
5.0 equivalents of sodium hydride in DMF was added 1.0 equivalent of N BOC-
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3-chloro-4-iodoaniline and then I.1 equivalents of iso-butyl 2-bromopropionate
(from Example AD above) were slowly added. The reaction was heated to
100°C for 10 hours, cooled, diluted with dichloromethane and washed
with cold
1N HCI, water and brine. The solvents were removed at reduced pressure and
S the residue was chromatographed to provide N BOC-N (3-chloro-4-
iodophenyl)alanine iso-butyl ester as a clear oil. Following General Procedure
AN above, the BOC group was removed from N BOC-N {3-chloro-4-
iodophenyl)alanine iso-butyl ester to provide the title compound. The BOC-
removal reaction was monitored by tlc on silica gel (Rf = 0.58 in 30%
EtOAc/hexanes) and purification was by preparative plate chromatography
(silica
gel using 30% EtOAc/hexanes as the eluant). The compound was further
purified by chromatography on an HPLC chiral column (Chiralcel OD).
NMR data was as follows:
'H-nmr (CDCI,): 8 = 7.52 (d, J=8.7, 1 H); 6.72 ( d, J=2.7, 1 H); 6.25 (dd,
J=2.7, 5.9, 2.7, 1 H); 4.35 (d, J=6.6, 1 H): 4.08 (quintex, J=7.2, 6.7, 1 H);
3.93 (d,
J=6.7, 2H): 1.94 (m, 1H); 1.47 (d, J=6.9, 3H); 0.92 (d, J=6.9, 6H).
'~C-nmr (CDC13): ~ = 174.5, 148.3, 140.7, 139.5, 114.4, I 14.3, 82.6,
72.0, 52.2, 28.3, 19.6, 19.3.
C,3H"C1IN0, (MW = 381.5); mass spectroscopy (MH+) 382.
Example A33
Synthesis of N-(4-azidophenyl)alanine iso-butyl ester
Following General Procedure AA above and using 4-azidoaniline
(Aldrich) and iso-butyl pyruvate (prepared by following General Procedure AO
above), the title compound was prepared as an oil. The reaction was monitored
by tlc on silica gel (Rf = 0.3 in 25% EtOAc/hexanes) and purification was by
preparative plate chromatography (silica gel using 25% EtOAc/hexanes as the
eluant).
NMR data was as follows:
'H-nmr (CDC13): 8 = 7.3 (d, 2H), 6.8 (d, 2H), 4.30 (bs, 1H), 4.08 (q,
1 H), 1.94 (sept, 1 H), I .47 (d, 3 H), 0.91 (d, 6H).
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''C-nmr (CDC13): 8 = 174.5, 148.7, 131.5, 130.3, 121.3, 1 14:9, 1 13.6,
72.0, 52.4, 28.3, 19.5, 19.3.
' C,3H;gN40, (MW = 262.31); mass spectroscopy (MH+) 263.
S Example A34
Synthesis of
N-[(4-phenylcarbonyl)phenylJalanine iso-butyl ester
Following General Procedure AA above and using 4'-
aminobenzophenone (Aldrich) and i.so-butyl pyruvate (prepared by following
General Procedure AO above), the title compound was prepared as an oil. The
reaction was monitored by tlc on silica gel (Rf = 0.4 in 25% EtOAc/hexanes)
and purification was by preparative plate chromatography (silica gel using 25%
EtOAc/hexanes as the eluant).
NMR data was as follows:
'H-nmr (CDCI,): 8 = 7.7 (d, 2H), 7.1 (m, SH), 6.9 (d, 2H), 4.30 (bs,
1 H), 4.08 (q, 1 H), 1.94 (sept, I H), 1.47 (d, 3H), 0.91 (d, 6H).
'''C-nmr (CDC13): b = 199, 178.5, 149.7, 131.5, 130.3, 126, 121.3, 114.9,
113.6, 72.0, 52.4, 28.3) 19.5, 19.3.
C,oH~3N03 (MW = 325.41 ); mass spectroscopy (MH+) 326.
Example A35
Synthesis of N-(3,5-difluorophenyl)alanine iso-butyl ester
Following General Procedure AH above and using N-(3,5-
difluorophenyl)alanine (from Example AC above) and iso-butanol, the title
compound was prepared as an oil. The reaction was monitored by tlc on silica
gel (Rf = 0.9 in 3% methanol/methylene chloride) and purification was by
preparative plate chromatography (silica gel using 3% methanollmethylene
chloride as the eluant).
NMR data was as follows:
. 30 'H-nmr (CDCI,): 8 = 6.1 (m, 3H), 4.5 (bs, 1H), 4.1 (d, 1H), 3.95 (m,
2H), 2.0 (m, 1H), 1.5 (d, J = 7 Hz, 3H), 0.95(d, J = 6 Hz, 6H).
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'''C-nmr (CDCl3}: 8 = 174.44, 166.40, 166.19, 163.16, 162.95, 149.43,
96.73, 96.60, 96.48, 96.35, 94.06, 93.72, 93.37, 72.03, 52.30, 28.29, 19.47,
19.23. -
C,3H,~F,N02 (MW = 290.2); mass spectroscopy (MH+) 291.
Example A36
Synthesis of N-(3,4-dichlorophenyl)alanine O-acylacetamidoxime ester
Following General Procedure AK above and using N-(3,4-
dichlorophenyl)alanine (from Example AB above) and acetamide oxime
(prepared according to the procedures described in J. Org. Chem., 46, 3953
( 1981 )), the title compound was prepared as a semisolid. The reaction was
monitored by tlc on silica gel (Rf = 0.4 in ethyl acetate) and purification
was by
preparative plate chromatography (silica gel using ethyl acetate as the
eluant).
NMR data was as follows:
'H-nmr (DMSO-db): 8 = 7.27 (d, 1H), 6.81 (s, 1H) 6.4 (broad s, 2H),
6.62 (d, 1 H), 6.45 (d, 1 H), 4.22 {m, 1 H), 1.74 (s, 3H), 1.40 (d, 3H).
C"H,3C1,N30~ (MW = 290.15); mass spectroscopy (MH~) 291.
Example A37
Synthesis of N-(3,4-dichlorophenyl)alanine pyrrolyl amide
Following General Procedure AL above and using N-(3,4-
dichlorophenyl)alanine (from Example AB above) and pyrrole (Aldrich), the
title
compound was prepared as an oil. The reaction was monitored by tlc on silica
gel (Rf = 0.28 in 10% ethyl acetate/hexanes) and purification was by
preparative
plate chromatography (silica gel using 10% ethyl acetate/hexanes as the
eluant).
NMR data was as follows:
'H-nmr (CDC1,): F~ = 7.36 (d, J=2.2, 2H); 7.20 (d, J=8.7, 1H); 6.71 (d,
J=2.7, 1 H); 6.5 (m, 1 H); 6.38 (t, J=2.4, 2H); 4.8 (m, 1 H); 4.57 {d, J=8.7,
1 H);
1.59 (d, J=6.8, 3H).
,..... ~ _..
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'3C-nmr (CDC13): 8 = 171.9, 146.1, 133.6, 131.5, 121.9, 119:6, 115.4,
' 114.7, 113.8, 51.8, 20.2.
C,3H,;ChN,O (MW = 283); mass spectroscopy (MH') 284.
Example A38
Synthesis of N-(3,4-dichlorophenyl)alanine O-acylbutyramidoxime ester
Following General Procedure AI above and using N-(3,4
dichlorophenyl)alanine 2,4,6-trichlorophenyl ester (prepared from N-{3,4
dichlorophenyl)alanine methyl ester (from Example A9) using essentially the
same procedure as described in Example AE above) and butyramide oxime
(prepared according to the procedures described in J. Org. Chem., 46, 3953
( 1981 )), the title compound was prepared as a semisolid. The reaction was
monitored by tlc on silica gel (Rf = 0.25 in 50% ethyl acetate/hexanes) and
purification was by preparative plate chromatography (silica gel using 50%
ethyl
acetate/hexanes as the eluant).
NMR data was as follows:
'H-nmr (db-DMSO): 8 = 7.27 (d, 1 H), 6.83 (s, 1 H) 6.38 (broad s, 2H),
6.61 (d, 1 H), 6.46 (d, 1 H), 4.25 (m, 1 H), 2.02 (t, 2H), 1.55 (m, 2H), 1.40
(d,
3H), 0.88 (t, 3H).
C,3H"C1,N30, (MW = 318.20); mass spectroscopy (MHi) 319.
Example A39
Synthesis of 2-[N-(naphth-2-yl)aminoJbutanoic acid ethyl ester
Following General Procedure AJ above and using 2-aminonaphthalene
(Aldrich) and ethyl 2-bromobutyrate (Aldrich), the title compound was prepared
as a solid, m.p. 81-83°C. The reaction was monitored by silica gel tle
(Rf = 0.5
in CHCl3). Purification was by chromatography (silica gel using chloroform as
the eluant).
NMR data was as follows: 'H-nmr (db-DMSO): 8 = 7.63 (m, 2H}, 7.54
(d, 1 H), 7.31 (t, 1 H), 7.12 (t, 1 H), 7.03 (d, 1 H), 6.62 (s, 1 H), 6.32 (d,
1 H), 4.15
(m, 3H), 1.42 (d, 3H), 1.19 (t, 3H).
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C,6H,9N0, (MW = 257.34); mass spectroscopy (MH+) 258. -
- Example A40
Synthesis of N-(2-naphthyl)alanine iso-butyl ester
Following General Procedure AA above and using 2-aminonaphthalene
(Aldrich) and iso-butyl pyruvate (prepared by following General Procedure AO
above), the title compound was prepared as an oil. Purification was by
preparative plate chromatography (silica gel using 25% EtOAc/hexanes as the
eluant).
NMR data was as follows:
'H-nmr (CDCI,): 8 = 7.65 (m, 3H), 7.38 (t, 1H, J = 6.9 Hz), 7.23 (t,
1 H, J = 6.9 Hz), 6.93 (m, 1 H), 6.81 (d, 1 H, J = 2.3 Hz), 4.31 (q, 1 H,
J = 6.9 Hz), 3.95 J = 6.7 Hz, J = 1.6 Hz), 1.96 (sept, 1 H, J = 6.7 Hz), 1.57
(d,
3H, J = 6.9 Hz), 0.93 (dd, 6H, J = 6.7 Hz, J = 1.6 Hz).
''C-nmr (CDCl3) 8 = 174.6, 144.2, 134.9, 129.1, 127.8, 127.6, 126.3,
126.0, 122.3, 118.1, 105.3, 71.2, 52.0, 27.7, 18.9, 18.8.
Example A41
Synthesis of N-(2-methylquinolin-6-yl)alanine iso-butyl ester
Following General Procedure AA above and using 6-amino-2-
methylquinoline (Lancaster) and iso-butyl pyruvate (prepared by following
General Procedure AO above), the title compound was prepared. The reaction
was monitored by silica gel tlc (Rf = 0.44 in 50% EtOAc/hexanes). Purification
was by flash chromatography (silica gel using 50% EtOAc/hexanes as the
eluant).
NMR data was as follows:
'H-nmr (CDCI,): b = 7.90 (m, 21-1), 7.10 (m, 2H), 6.66 (d, 1H, J = 2.6),
4.50 (bd, 1 H), 4.24 (m, 1 H), 3.91 (d, 2H, 3 = 6.6 Hz), 2.64 (s, 3H), 1.91
(sept,
1H, J = 6.7 Hz), 1.52 (d, 3H, J = 6.9 Hz), 0.87 (d, 6H, J = 6.7 Hz).
'3C-nmr (CDCl3) ~ = 175.0, 155.4, 144.6, 143.4, 134.9, 130.2, 128.4,
. 122.8, 121.8, i 04.9, 71.8, 52.7, 28.3, 25.4, 19.5, 19.4.
...,.......»"..... . 1,
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C"H~,CI~N,O~ {MW = 286.38); mass spectroscopy (MH') 287.
" - Example A42
Synthesis of N-(3,4-methylenedioxyphenyl)alanine iso-butyl ester
Following reductive amination General Procedure AA above and using
3,4-methylenedioxyaniline (Aldrich) and methyl pyruvate (Aldrich), N-(3,4-
methylenedioxyphenyl)alanine methyl ester was prepared. The methyl ester was
then transesterified following General Procedure AQ above and using iso-
butanol to provide the title compound as an oil. The reaction was monitored by
silica gel tlc (Rf = 0.61 in 25% EtOAclhexanes). Purification was by
preparative plate chromatography with silica gel using 25% EtOAc/hexanes as
the eluant.
NMR data was as follows:
'H-nmr (CDCI,): d = 6.63 (d, 1H, 8.3 Hz), 6.25 (d, 1H, J = 2.3 Hz),
6.04 (dd, 1 H, J = 8.3 Hz, ,1 = 2.3 Hz), 5.83 (s, 2H), 3.96 (m, 4H), 1.92
(sept,
1 H, J = 6.7 Hz), 1.44 (d, 3 H, J = 6.9 Hz), 0.90 (d, 6H, J = 6.6 Hz).
'3C-nmr (CDCl3): 8 = 175.4, 148.9, 142.9, 140.8, 109.2, 105.8, 101.2,
97.4, 71.6, 53.6, 28.3, 19.6, 19.5.
C,,,H,9N04 (MW = 265.31 ); mass spectroscopy (MH') 265.
Example A43
Synthesis of N-(3,4-ethylenedioxyphenyl)alanine iso-butyl ester
Following reductive amination General Procedure AA above and using
1,4-benzodioxa-6-amine (Aldrich) and methyl pyruvate (Aldrich), N-(3,4-
ethylenedioxyphenyl)alanine methyl ester was prepared. The methyl ester was
then transesterified following General Procedure AQ above using iso-butanol to
provide the title compound. Purification was by preparative plate
chromatography.
NMR data was as follows:
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'H-nmr (CDCI,): 8 = 0.91 (d, J = 7Hz; 6H), 1.42 (d, J = 7Hz, 3H), 1.8-
2.0 (m, 1 H), 3.8-3.95 (m, 3H), 4.0-4.1 (m, 1 H), 4.15-4.25 (m, 4H), 6.12-6.2
(m,
2H), 6.65-6.75 (m, 1H).
''C-nmr (CDCl3): 8 = 19.55, 19.56, 19.67, 28.3, 53.4, 64.7, 65.3, 71.7,
103.1, 108.0, 118.3, 142.1, 144.6, 175.4.
C,SH,,N04 (MW = 279.34); mass spectroscopy (MH+) 280.
Example A44
Synthesis of N-(2-naphthyl)alanine methyl ester
Following reductive amination General Procedure AA above and using 2-
aminonaphthalene (Aldrich) and methyl pyruvate (Aldrich), the title compound
was prepared. The reaction was monitored by silica gel tlc (Rf = 0.50 in 25%
EtOAclhexanes). Purification was by flash chromatography with silica gel using
25% EtOAclhexanes as the eluant.
NMR data was as follows:
'H-nmr (CDCI,): 8 = 7.65 (m, 3H), 7.48 (m, 1 H), 7.25 (m, 1 H), 6.91
(m, 1H), 6.79 (m, 1H), 4.31 (m, 2H), 3.76 (s, 3H), 1.55 (d, 3H).
'3C-nmr (CDC13): b = 175.66, 144.78, 135.55, 129.78, 128.47, 128.22,
126.96, 126.67, 123.01, 118.66, 105.88, 52.95, 52.51, 19.45.
C,4H,5N02 (MW = 229.28); mass spectroscopy (MH+} 229.
Example A45
Synthesis of N-(benzothiazol-6-yl)alanine ethyl ester
To a solution of 6-aminobenzothiazole (Lancaster} in dichloromethane
was added 1.2 equivalents of pyridine, followed by 1.5 equivalents of
trifluoroacetic anhydride. The reaction was stirred at room temperature for 3
hours and then washed with 5% citric acid, dried over MgS04, and stripped free
of solvent on a rotary evaporator to yield 6-trifluoroacetamidothiazole. This
material was dissolved in THF and then added to a suspension of KH in THF at
0°C. A catalytic amount of 18-crown-6 was added, followed by ethyl 2-
bromopropionate (Aldrich). The reaction was held at room temperature for 1
_..r.__.~.._r.~.,.....e..~....___... ,
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hour, and then heated to reflux for 24 hours, and then cooled to room
temperature. The reaction mixture was stripped free of solvent on a rotary
' evaporator 'and the resulting residue was dissolved in ether. This solution
was
washed with water, saturated aqueous NaCI, and dried over MgS04. The
solution was stripped free of solvent on a rotary evaporator and the title
compound was obtained by chromatography of the residue using 5%
methanol/dichloromethane (Rf = 0.59) as the eluant.
NMR data was as follows:
'H-nmr (CDCI,): 8 = 8.69 (s, 1H), 7.90 (d, IH, J = 8.8 Hz), 7.04 (d,
1 H, J = 2.3 Hz}, 6.84 (dd, 1 H, J = 8.8 Hz, J = 2.4 I ~z), 4.41 (bd, I H, J =
7.5
Hz), 4.20 (m, 3H), 1.53 (d, 3H, J = 6.9 Hz), 1.27 (t, 3H, J = 7.1 Hz).
'3C-nmr (CDC13): ~ = 174.9, 150.2, 147.1, 145.6. 136.3, 124.6, 115.7,
103.5, 61.9, 52.9, 19.4, 14.8.
C,,H,4N~O,S (MW = 250.32); mass spectroscopy (MI-I') 251.
Example A46
Synthesis of N-(indol-5-yl)alanine iso-butyl ester (S isomer)
Following General Procedure AM and using 5-aminoindole (Aldrich) and
iso-butyl R-(+)-lactate (Aidrich), the title compound was prepared as an oil.
The reaction was monitored by silica gel tlc (Rf = 0.46 in 33% EtOAc/hexanesj.
Purification was by preparative plate chromatography with silica gel using 33%
EtOAc/hexanes as the eluant.
NMR data was as follows:
'H-nmr (CDCI,): b = 8.11 (bs, 1 >-I), 7.07 (d, J=8.8 Hz, I H), 6.98 (d,
J=2.8 Hz, 1 H), 6.83 (d, J=2.2 Hz, 1 H), 6.61 (m, 1 H), 6.32 (m, 1 H), 4.18
(q,
J=6.9 Hz, I H), 3.95 (bs, 1 H), 3.87 (d, J=6.7 Hz, 2H), 1.89 (hept, J=6.7 Hz,
1 H},
- 1.48 (d, J=6.96 Hz, 3H), 0.86 (dd, J=6.7 Hz, J=1.6 Hz, 6H).
'3C-nmr (CDC1~): b = 176.15, 141.06, 131.28, 129.24, 125.34, I 13.34,
112.53, 104.21, 102.17, 71.65, 54.28, 28.36, 19.87, 19.62.
C,SH,°NzO, {MW = 260.34); mass spectroscopy (MH+) 261.
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Example A47 -
Synthesis of N-(naphth-2-yl)alanine O-acylacetamidoxime ester
Following General Procedure AI above using N-(naphth-2-yl)alanine
2,4,6-trichlorophenyl ester (from Example AE above) and acetamide oxime
(prepared according to the procedures described in J. Org. Chem. , 46, 3953
( 1981 )), the title compound was prepared as a semisolid. The reaction was
monitored by tlc on silica gel (Rf = 0.4 in ethyl acetate) and purification
was by
preparative plate chromatography (silica gel using ethyl acetate as the
eluant).
NMR data was as follows:
' H-nmr (dfi-DMSO): S = 7.64 (t, 2H), 7.54 (d, 1 H), 7.32 (t, 1 H), 7.13 (t,
1 H), 7.04 (d, 1 H), 6.78 (s, 1 H) 6.42 (broad s, 2H), 6.32 (d, 1 H), 4.33 (m,
1 H),
1.72 (s, 3H), 1.46 (d, 3H).
CjSH,~N3O, (MW = 271.32); mass spectroscopy: 271.
Example A48
Synthesis of N-(2-naphthyl)alanine ethyl ester
Following reductive amination General Procedure AA above and using 2-
aminonaphthalene (Aldrich) and ethyl pyruvate {Aldrich), the title compound
was prepared as a solid having a melting point of 52-56°C. The reaction
was
monitored by silica gel tlc (Rf = 0.50 in 25% EtOAc/hexanes). Purification was
by flash chromatography with silica gel using 25% EtOAc/hexanes as the eluant.
NMR data was as follows:
'H-nmr (CDC13): 8 = 7.65 (m, 3H), 7.48 (m, 1H), 7.25 (m, 1H), 6.91
(m, 1H), 6.79 (m, iH), 4.31 (m, 2H), 3.76 (s, 3H), 1.55 (d, 3H).
'3C-nmr (CDC13): 8 = 175.66, 144.78, 135.55, 129.78, 128.47, 128.22,
126.96, 126.67, 123.01, 118.66, 105.88, 52.95, 52.51, 19.45.
C,qH,5N0, (MW = 229.28); mass spectroscopy (MH+) 229.
Example A49
Synthesis of N-(3,4-dichlorophenyl)alanine O-acylpropionamidoxime ester
_.. ._ ~...-.,....._ a .. .. .. . r
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Following General Procedure AI above using N-(3,4- -
' dichlorophenyl)alanine 2,4,6-trichlorophenyl ester (prepared from N-(3,4-
dichlorophenyl)alanine methyl ester (from Example A9) using essentially the
same procedure as described in Example AE above) and propionamide oxime
' S (prepared according to the procedures described in J. Org. Chem., 46, 3953
( 1981 )), the title compound was prepared as a semisolid. The reaction was
monitored by tlc on silica gel (Rf = 0.2 in 50% ethyl acetate/hexane) and
purification was by preparative plate chromatography (silica gel using 50%
ethyl
acetate/hexane as the eluant).
NMR data was as follows:
'H-nmr (d''-DMSO): 8 = 7.27 (d, 1 H), 6.83 (s, 1 H), 6.64 (d, 1 H), 6.47 (d,
1H), 6.38 (broad s, 2H), 4.24 (m, 1H), 2.07 (q, 2H), 1.41 (d, 3H).
C,~H,SCI,N30, (MW = 304.17); mass spectroscopy (MH') 305.
Example A50
Synthesis of N-(4-ethoxycarbonylphenyl)alanine iso-butyl ester (S isomer)
Following General Procedure AM and using ethyl 4-aminobenzoate
(Aldrich) and iso-butyl R-{+)-lactate (Aldrich), the title compound was
prepared
as an oil. The reaction was monitored by silica gel tlc (Rf = 0.21 in 10%
EtOAc/hexanes). Purification was by preparative plate thin layer
chromatography using 25% EtOAc/hexanes as the eluant.
NMR data was as follows:
'H-nmr (CDCI,): 8 = 7.82 (d, J = 8.73 Hz, 2H), 6.51 (d, J = 8.79 Hz,
2H), 4. 81 {d, J = 7.82 Hz, 1 H), 4.25 (q, J = 7.14 Hz, 2H), 4.15 (quint, J =
7.40
Hz, 1 H), 3.87 (m, 2H), 1.87 (sept, J = 6.70 Hz, 1 H), 1.43 (d, J = 6.95 Hz, 3
H),
1.30 (t, J = 7.14 Hz, 3H), 0.84 (d, J = 6.71 Hz, 6H).
- ~'C-nmr (CDC13): 8 = 174.5, 167.3, 151.0, 132.0, 119.9, 112.5, 71.9,
60.8, 51.9, 28.2, 19.5, 19.2, 15Ø
C,6H~3N04 {MW = 293.37); mass spectroscopy (MH+) 294.
Example A51
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Synthesis of N-[3,S-di(triftuoromethyl)phenyl]alanine iso-butyl
ester (S isomer)
Following General Procedure AM and using 3,5-
di(trifluoromethyl)aniline (Aldrich) and isa-butyl R-(+)-lactate (Aldrich),
the title
compound was prepared as an oil. The reaction was monitored by silica gel tlc
(Rf = 0.38 in 10% EtOAc/hexanes). Purification was by preparative plate thin
layer chromatography using 10% EtOAc/hexanes as the eluant.
NMR data was as follows:
'H-nmr (CDCI,): 8 = 7.13 (s, IH), 6.91 (s, 2H), 4.97 {d, J = 8.24 Hz,
1 H), 4. I 8 (m, 1 H), 3.93 (d, J = 6.59 Hz, 2H), I .93 (sept, 3 = 6.7 I Hz, 1
H), 1.49
(d, J = 7.02 Hz, 3H), 0.89 (d, J = 6.59 Hz, 6H).
''C-nmr (CDC13): 8 = 174.4, 147.9, 133.6, 133.2, 132.7, 132.3, 129.4,
125.8, 122.2, 118.6, 112.81, 112.76, 111.42, III.37, 111.32, 111.27, 111.22,
72.2, 52.0, 32.1, 28.24, 28.17, 23.2, 19.5, 19.3, 19.2, 18.9, 14.6.
C'5H"F6N0~ (MW = 357.30); mass spectroscopy (MH+) 358.
Example A52
Synthesis of N-(3,5-dimethoxyphenyl)alanine iso-butyl
ester
N-(3,5-dimethoxyphenyl)alanine (crude, 454 mg) (prepared according to
the procedure described in U.S. Patent No. 3,598,859 using 3,5-
dimethoxyaniline
(Aldrich) and 2-chloropropionic acid (Aldrich)) was treated in dry iso-butanol
( 10 mL) with 0.1 mL of chlorotrimethylsilane and the reaction mixture
refluxed
overnight. The excess alcohol was removed at reduced pressure and the residue
dissolved in ethyl acetate. The ethyl acetate solution was washed with
saturated
aqueous NaHC03, dried with Na.,S04 and the solvent removed to provide the
title compound. The reaction was monitored by silica gel tlc (Rf = 0.3 in 20%
EtOAc/hexanes). Purification was by preparative thin layer chromatography
using 20% EtOAc/hexanes as the eluant.
NMR data was as follows:
__......_ ...~,_-..~.~.. --...",....... ~
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'H-nmr (CDCI,): 8 = 0.9 {d, J = 7, 6H), 1.47 (d, J = 7, 3H); 1.9-2.0 (m,
1 H), 3.7 (s, 6H), 3.85-4.0 (m, 2H), 4.1-4.2 (m, 1 H), 4.3 (brs, 1 H), 5.8 (s,
2H),
5.9 (s, 1 H}. -
''C-nmr (CDCI,): 8 = 19.49, 19.52, 19.54, 28.3, 52.5, 55.6, 71.7, 91.1,
92.7, 149.2, 162.3, 175.2.
C,SH,3N04 (MW = 281.35}.
Example A53
Synthesis of N-(2-napthyl)alanine O-acylpropionamidoxime ester
Following General Procedure AS and using N-(2-naphthyl)alanine 2,4,5-
trichlorophenyl ester (from Example AE above) and propionamide oxime
(prepared according to the procedures described in J. Org. Chem. , 46, 3953
( 1981 )), the title compound was prepared. The reaction was monitored by
silica
gel tlc (Rf = 0.5 in EtOAc). Purification was by silica gel chromatography
using 1:1 EtOAc/hexanes as the eluant.
NMR data was as follows:
'H-nmr (DMSO-db): 8 = 1.03 (t, 3H), 1.45 (d, 3H).
C,6H,9N302 (MW = 285.35); mass spectroscopy (M~) 285.
Example A54
Synthesis of N-(2-napthyl)alanine O-acylbutyramidoxime ester
Following General Procedure AS and using N-(2-naphthyl)alanine 2,4,5-
trichlorophenyl ester (from Example AE above) and butyramide oxime (prepared
according to the procedures described in J. Org. Chem. , 46, 3953 ( 1981 )),
the
title compound was prepared as an oil. The reaction was monitored by silica
gel
tlc (Rf = 0.6 in EtOAc). Purification was by silica gel chromatography using
l:l EtOAc/hexanes as the eluant.
NMR data was as follows:
'H-nmr (DMSO-dn): 8 = 0.86 (t, 3H), 1.46 (d, 3H).
C"H~,N30, (MW = 299.37); mass spectroscopy (MH+) 299.
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Example A55 - -
Synthesis of N-(2-napthyl)alanine O-acylisovaleramidoxime ester
Following General Procedure AS and using N-{2-naphthyl)alanine 2,4,5-
trichlorophenyl ester (from Example AE above) and isovaleramide oxime
(prepared according to the procedures described in J. Org. Chem. , 46, 3953
( 1981 )), the title compound was prepared as an oil. The reaction was
monitored
by silica gel tlc (Rf = 0.3 in 1:1 EtOAc/hexanes). Purification was by silica
gel
chromatography using 1:1 EtOAc/hexanes as the eluant.
NMR data was as follows:
'H-nmr (DMSO-dn): 8 = 0.86 (t, 3H), 1.45 (d, 3H).
C,8H,3N30, (MW = 313.40); mass spectroscopy (MH+) 313.
Example A56
Synthesis of N-(2-napthyl)alanine O-acylbenzamidoxime ester
Following General Procedure AS and using N-(2-naphthyl)alanine 2,4,5-
trichlorophenyl ester (from Example AE above} and benzamide oxime (prepared
according to the procedures described in J. Org. Chem., 46, 3953 (1981)), the
title compound was prepared as an oil. The reaction was monitored by silica
gel
tlc (Rf = 0.3 in 1:1 EtOAc/hexanes). Purification was by silica gel
chromatography using 1:1 EtOAc/hexanes as the eluant.
NMR data was as follows:
'H-nmr (DMSO-db): S = 4.42 (m, 1H), 1.53 (d, 3H).
CZOH,9N302 (MW = 333.39}; mass spectroscopy (MH') 333.
Example A57
Synthesis of N-(2-napthyl)alanine O-acylcyclopropanecarboxamidoxime
ester
Following General Procedure AS and using N-(2-naphthyl)alanine 2,4,5-
trichlorophenyl ester (from Example AE above) and cyclopropanecarboxamide
oxime (prepared according to the procedures described in J. Org. Chem. , 46,
3953 ( 1981 )), the title compound was prepared as an oil. The reaction was
....._......._.._~~....~.:."...~..,.,."",.",. "............ ~ . .,, ....... .
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monitored by silica gel tlc (Rf = 0.3 in 1:1 EtOAc/hexanes). Purification was
' by silica gel chromatography using 1:1 EtOAc/hexanes as the eluant.
NMR data was as follows:
'H-nmr (DMSO-d~): S = 0.85 (m, 4H), 1.43 (d, 3H).
C"H,9N30, (MW = 297.36); mass spectroscopy (MH') 297.
Example A58
Synthesis of N-(2-napthyl)alanine O-acylcyclopropylacetamidoxime ester
Following General Procedure AS and using N-(2-naphthyl)alanine 2,4,5-
trichlorophenyl ester (from Example AE above) and cyclopropylacetamide oxime
(prepared according to the procedures described in .l. Org. Chem., 46, 3953
( 1981 )), the title compound was prepared as an oil. The reaction was
monitored
by silica gel tlc (Rf = 0.3 in 1:1 EtOAc/hexanes}. Purification was by silica
gel
chromatography using I :I EtOAc/hexanes as the eluant.
NMR data was as follows:
'H-nmr (DMSO-dfi): 8 = I.43 (d, 3H), 1.91 (d, 2H).
C,$H.,,N30, (MW = 311.39); mass spectroscopy (MH') 311.
Example A59
Synthesis of N-(2-napthyl)alanine O-acylcyclopentanecarboxamidoxime
ester
Following General Procedure AS and using N-(2-naphthyl)alanine 2,4,5-
trichlorophenyl ester {from Example AE above) and cyclopentanecarboxamide
oxime (prepared according to the procedures described in J. Org. Chem. , 46,
3953 ( 1981 )), the title compound was prepared as an oil. The reaction was
monitored by silica gel tlc (Rf = 0.3 in 1:1 EiOAc/hexanes). Purification was
by silica gel chromatography using 1:1 EtOAc/hexanes as the eluant.
NMR data was as follows:
'H-nmr (DMSO-d~): 8 = 1.43 (d, 3H), 2.43 (m, 1H).
' C"H,9N30, (MW = 297.36).
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GENERAL PROCEDURE BA
Coupling of R'C(X'~X")C(O)Cl with HzNCH(R'-~C(O)XR3
To a-stirred solution of (D,L)-alanine iso-butyl ester hydrochloride (from
Example BB below) (4.6 mmol) in 5 mL of pyridine was added 4.6 mmol of an
acid chloride. Precipitation occurred immediately. The mixture was stirred for
3.5 h, diluted with 100 mL of diethyl ether, washed with 10% HCI three times,
brine once, 20% potassium carbonate once and brine once. The solution was
dried over magnesium sulfate, filtered, and evaporated at reduced pressure to
yield the product. Other amino acid esters may also be employed in this
procedure.
GENERAL PROCEDURE BB
Coupling of R'C(X')(X")C(O)OH with HZNCH(R'-)C(O)XR3
A solution of the acid (3.3 mmol) and CDI in 20 mL THF was stirred
for 2 h. L-alanine iso-butyl ester hydrochloride (from Example BB below) (3.6
mmol) was added, followed by 1.5 mL (10.8 mmol) of triethylamine. The
reaction mixture was stirred overnight. The reaction mixture was diluted with
100 mL of diethyl ether, washed with 10% HC1 three times, brine once, 20%
potassium carbonate once and brine once. The solution was dried over
magnesium sulfate, filtered, and evaporated at reduced pressure to yield the
product. Other amino acid esters may also be employed in this procedure.
GENERAL PROCEDURE BC
Esterification of R'C(X')(X")C(O~NHCH~R'-)C(O)OH With HORS
To a stirred solution of phenylacetyivaline ( 1.6470 g, 7.0 mmol) in 20
mL THF was added CDI ( 1.05 g, 6.5 mmol) and the mixture was stirred for 1.5
h. 2-Methylbutanol (0.53 g, 6 mmol) was added the mixture, followed by
addition of NaH (0.16 g, 6.5 mmol). Bubbling occurred immediately. The
reaction mixture was stirred overnight. The reaction mixture was diluted with
100 mL of diethyl ether, washed with 10% HCl three times, brine once, 20%
potassium carbonate once and brine once. The solution was dried over
,.
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magnesium sulfate, filtered, and evaporated at reduced pressure to yield the
product. Other N-acyl amino acids and alcohols may also be employed in this
procedure. -
GENERAL PROCEDURE BD
Ester Hydrolysis to the Free Acid
Ester hydrolysis to the free acid was conducted by conventional methods.
Below are two examples of such conventional de-esterification methods.
To the ester in a 1:1 mixture of CH,OH/H>O was added 2-5 equivalents
of K,CO:. The mixture was heated to about 50°C for about 0.~ to 1.5
hours
until tlc showed complete reaction. The reaction was cooled to room
temperature and the methanol was removed at reduced pressure. The pH of the
remaining aqueous solution was adjusted to about 2, and ethyl acetate was
added
to extract the product. The organic phase was then washed with saturated
aqueous NaCI and dried over MgS04. The solution was stripped free of solvent
at reduced pressure to yield the product.
The amino acid ester was dissolved in dioxanelwater (4:1 ) to which was
added LiOH (~2 eq.) that was dissolved in water such that the total solvent
after
addition was about 2:1 dioxane:water. The reaction mixture was stirred until
reaction completion and the dioxane was removed under reduced pressure. The
residue was diluted with EtOAc, the layers were separated and the aqueous
layer
acidified to pH 2. The aqueous layer was back extracted with EtOAc, the
combined organics were dried over Na,SO~ and the solvent was removed under
reduced pressure after filtration. The residue was purified by conventional
. methods (e.g., recrystallization).
The following exemplifies this later example. The methyl ester of 3-NO,
phenylacetyl alanine 9.27 g (0.0348 mots) was dissolved in 60 mL dioxane and
15 mL of H,O and adding LiOH (3.06 g, 0.0731 mol) that has been dissolved in
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15 mL of HBO. After stirring for 4 hours, the dioxane was removed under
reduced pressure and the residue diluted with EtOAc, the layers were separated
and the aqueous layer acidified to pH 2. The aqueous layer was back extracted
with EtOAc (4 X 100 mL}, the combined organics were dried bver Na,S04 and
the solvent was removed under reduced pressure after filtration. The residue
was recrystallized from EtOAc/isooctane giving 7.5 g (85%) of 3-
nitrophenylacetyl alanine. C, ~ H,,N~OS requires C = 52.3 8, H = 4.80, and N =
11.11. Analysis found C = 52.54, H = 4.85, and N = 11.08. [a],3 = - 29.9 @
589 nm.
GENERAL PROCEDURE BE
Low Temperature BOP Coupling of Acid and Alcohol
A solution of methylene chloride containing the carboxylic acid
( 100M%) and N-methyl morpholine ( 150 M%) was cooled to -20°C under
nitrogen. BOP ( 1 OS M%) was added in one portion and the reaction mixture
was maintained at -20°C for 15 minutes. The corresponding alcohol ( 120
M%)
was added and the reaction mixture was allowed to warm to room temperature
and stirred for 12 hours. The reaction mixture was then poured into water and
extracted with ethyl acetate (3x}. The combined ethyl acetate portions were
backwashed with saturated aqueous citric acid (2x), saturated aqueous sodium
bicarbonate (2x), brine ( 1 x), dried over anhydrous magnesium sulfate or
sodium
sulfate and the solvent removed under reduced pressure to yield the crude
product.
GENERAL PROCEDURE BF
EDC Coupling of Acid and Amine
The acid derivative was dissolved in methylene chloride. The amine
( 1 eq.), N-methylmorpholine (5 eq.), and hydroxybenzotriazole monohydrate
( 1.2 eq.) were added in sequence. The reaction was cooled to about 0°C
and
then 1.2 eq. of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride
was added. The solution was allowed to stir overnight and come to room
_. ...__ ..~.. ,
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temperature under N, pressure. The reaction mix was worked up by washing the
solution with saturated, aqueous Na.:CO" 0.1 M citric acid, and brine before
' drying with Na,S04 and removal of solvents to yield crude product. Pure
products were obtained by flash chromatography in an appropriate solvent.
' S
GENERALPROCEDURE BG
EDC Coupling of Acid and Amine
A round bottom flask was charged with carboxylic acid (1.0 eq.),
hydroxy-benzotriazole hydrate ( 1.1 eq.) and amine ( 1.0 eq.) in THF under
nitrogen atmosphere. An appropriate amount ( 1.1 eq. for free amines and 2.2
eq. for hydrochloride amine salts) of base, such as Hunig's base was added to
the well stirred mixture followed by EDC ( 1.1 eq. j. After stirring from 4 to
17
hours at room temperature the solvent was removed at reduced pressure, the
residue taken up in EtOAc (or similar solvent)Iwater. The organic layer was
washed with saturated aqueous sodium bicarbonate solution, 1N HCI, brine and
dried over anhydrous sodium sulfate. In some cases, the isolated product was
analytically pure at this stage while, in other cases, purification via
chromatography and/or recrystallization was required prior to biological
evaluation.
GENERAL PROCEDURE BH
Coupling of R'C(X')(X")L(O)CI with HzNCH(R'1C(O)XR3
An excess of oxalyl chloride in dichloromethane was added to the acid
derivative together with one drop of DMF . The resulting mixture was stirred
for
about 2 hours or until bubbling ceases. The solvent was then removed under
reduced pressure and rediluted with dry methylene chloride. To the resulting
solution was added about 1.1 eq. of the appropriate amino acid ester and
triethylamine ( 1.1 eq. in methylene chloride). The system was stirred at room
temperature for 2 hours and then the solvent was removed under reduced
pressure. The residue was dissolved in ethyl acetate, washed with IN HCl
followed by 1 N NaOH. The organic layer was dried over anhydrous soldium
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sulfate, filtered and the solvent removed under reduced pressure to provide
for
the desired product.
GENERAL PROCEDURE BI -
P-EPC coding
P-EPC coupling employs an amino acid ester and a substituted acetic
acid compound. The acetic acid derivative is well known in the art and is
typically commercially available. The amino acid ester is prepared by
conventional methods from the known and typically commercially available N-
BOC amino acid as described in GENERAL PROCEDURE BJ below.
Specifically, the appropriate amino ester free base (0.0346 mmols) and
substituted phenyiacetic acid (0.069 mmois) were dissolved in 2.0 mL CHCi3
(EtOH free), treated with 150 mg of P-EPC (0.87 meq./g) and the reaction was
mixed for 4 days at 23 °C. The reaction was filtered through a plug of
cotton,
rinsed with 2.0 mL of CHC13 and the filtrate evaporated under a stream of
nitrogen. The purity of each sample was determined by 'H NMR and ranged
from 50% to > 95 % . Between 8.0 and 15.0 mg of final product was obtained
from each reaction and was tested without additional purification.
GENERAL PROCEDURE BJ
Synthesis of Amino Acid Esters From the Corresponding N-BOC Amino Acid
A. Esterification of the Acid.
The N-BOC amino acid was dissolved in dioxane and treated with an
excess of alcohol (--1.5 eq.} and catalytic DMAP (100 mg) at 0°C.
Stirring
was continued until reaction completion whereupon the product was recovered
by conventional methods.
B. Removal of N-BOC Group.
The N-BOC protected amino acid was dissolved in methylene chloride
(O.OSM) and treated with 10 eq. of TFA at room temperature under a nitrogen
~___......~...._,..,?.,r.. -..~n....~.._ . .,_
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atmosphere. The reaction was monitored by tlc until starting material was
consumed usually within 1-5 hours. An additional 10 eq. of TFA was added to
the reaction if the starting material was still present after 5 hours. The
reaction
was carefully neutralized with NazC03, separated, the organic Dyer washed
with brine and dried over anhydrous NazS04. The crude amine was then used
without purification.
Specific exemplification of these procedures are as follows:
1. Racemic (+/-)-N-BOC-a-amino butyric acid (Aldrich) {9.29 g,
0.0457 mol) was dissolved in 100 mL of dioxane and treated with iso-butyl
alcohol (6.26 mL, 0.0686 mol), EDC {8.72 g, 0.0457) and catalytic DMAP
(100 mg) at 0°C. After stirring for 17 hours, the organics were
evaporated at
reduced pressure, the residue diluted with EtOAc washed with NaHC03, brine
and dried over NazS04. Evaporation yields 8.42 g (71 % ) of an oil. C,3Hz5NO4
requires: C = 60.21, H = 9.72, and N = 5.40. Anal found: C = 59.91,
H = 9.89, and N = 5.67.
The above N-BOC amino acid ester (8.00 g, 0.032 mol) was deprotected
as above giving 3.12 g (61 % ) of the free base as a colorless oil which
solidifies
upon standing.
2. L-N-BOC-alanine (Aldrich). (8.97 g, 0.047 mol) was dissolved in
100 mL of CHZCIz, iso-butyl alcohol (21.9 mL, 0.238 mol) and treated with
DMAP (100 mg) and EDC (10.0 g, 0.52 mol) at O°C. The mixture was
stirred for 17 hours, diluted with HzO, washed with 1.0 N HCI, NaHC03, then
brine and the organics were dried over NazS04. Filtration and evaporation
yields 11.8 g (quantitative) of L-N-BOC alanine iso-butyl ester which is
contaminated with a small amount of solvent. A sample was vacuum dried for
analytical analysis. C,zHz3NO4 requires: C = 58.79, H = 9.38, and N =
5.71. Anal found: C = 58.73, H = 9.55, and N = 5.96.
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The above N-BOC amino acid ester ( 11.8 g, 0.0481 mol) was
deprotected as above. The free base was converted to the corresponding HCI
salt using safurated HCl (g)/EtOAc to give L-N-alanine iso-butyl ester
hydrochloride. Obtained 4.2 g (48%) of a colorless solid. C.,H,5N02. HCI
requires:
C = 46.28, H = 8.88, and N = 7.71. Anal found: C = 46.01, H = 8.85,
and N = 7.68.
GENERAL PROCEDURE BK
Methyl ester formation from amino acids
The amino acid (amino acid or amino acid hydrochloride) is suspended
in methanol and chilled to 0°C. HCI gas is bubbled through this
solution for 5
minutes. The reaction is allowed to warm to room temperature then stirred for
4 hours. The solvents are then removed at reduced pressure to afford the
desired amino acid methyl ester hydrochloride. This product is usually used
without further purification.
Example BA
Synthesis of free and polymer bound PEPC
N-ethyl-N'-3-ll-nvrrolidinvl)nronvlurea
To a solution of 27.7 g (0.39 mol) ethyl isocyanate in 250 mL
chloroform was added 50 g (0.39 mol) 3-(1-pyrrolidinyl)propylamine dropwise
with cooling. Once the addition was complete, the cooling bath was removed
and the reaction mixture stirred at room temperature for 4 hours. The reaction
mixture was then concentrated under reduced pressure to give 74. 5 g (96.4 % )
of the desired urea as a clear oil.
1-(3-(1-p~rrrolidinyl)prowl)-3-ethylcarbodiimide (P-EPC)
To a solution of 31.0 g (0.156 mol) N-ethyl-N'-3-(1-pyrrolidinyl)propyl
urea in 500 mL dichloromethane was added 62.6 g (0.62 mol) triethylamine and
the solution was cooled to 0 ° C . To this solution were then added
59.17 g (0. 31
r
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mol) 4-toluenesulfonyl chloride in 400 mL dichloromethane dropwise at such a
rate as to maintain the reaction at 0-5°C. After the addition was
complete, the
reaction mixture was warmed to room temperature and then heated to reflux for
4 hours. After cooling to room temperature, the reaction mixture was washed
' S with saturated aqueous potassium carbonate (3 x 150 mL). The aqueous
phases
were combined and extracted with dichloromethane. All organic phases were
combined and concentrated under reduced pressure . The resultant orange slurry
was suspended in 250 mL diethyl ether and the solution decanted off from the
solid. The slurryldecantation process was repeated 3 more times. The ether
solutions were combined and concentrated under reduced pressure to give 18.9
g (67 % ) of the desired product as a crude orange oil. A portion of the oil
was
distilled under vacuum to give a colorless oil distilling at 78-82 °C
(0.4 mm
Hg).
Preparation of a polymer supported form of 1-(3-(I-Qyrrolidinyl)propyl)-3-
ethylcarbodiimide (P-EPC)
A suspension of 8.75 g (48.3 mmol) 1-(3-(1-pyrrolidin-yl)propyl)-3-
ethylcarbodiimide and 24. I7 g (24.17 mmol) Merrifield's resin (2 % cross-
linked, 200-400 mesh, chloromethylated styrene/divinylbenzene copolymer, 1
meq. Cl/g) in dimethylformamide was heated at 100°C for 2 days. The
reaction was cooled and filtered and the resulting resin washed sequentially
with
1L DMF, 1L THF and 1L diethyl ether. The remaining resin was then dried
under vacuum for 18 hours.
Example BB
Preparation of alanine iso-butyl ester hydrochloride
A mixture of 35.64 g (0.4 mol) of (D,L)-alanine (Aldrich) (or L-alanine
(Aldrich)); 44 mL (0.6 mol) of thionyl chloride (Aldrich) and 200 mL of
isobutanol was refluxed for 1.5 hours and the volatiles were removed
completely on a rotavapor of 90°C under reduced pressure to give (D,L)-
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alanine iso-butyl ester hydrochloride (or L-alanine iso-butyl ester -
hydrochloride), which was pure enough to be used for further transformations.
Example BC -
Preparation of 3,5-dichlorophenylacetic acid
To a solution of 3.5 g of 3,5-dichlorobenzyl alcohol (Aldrich) in 75 mL
of dichloromethane at 0°C was added 1.8 mL of methane sulfonylchloride
followed by 3.5 mL of triethylamine added dropwise. After 2 hours the
solution was diluted to 150 mL with dichioromethane, washed with 3N HCI,
saturated aqueous NaHC03 dried with Na,S04 and the solvents removed to
yield the desired 3,5-dichlorobenzyl methanesulfonate as a yellow oil that was
used without purification.
The crude sulfonate was dissolved in 50 mL of DMF at 0°C and then 3
g of KCN was added. After 2 hours an additional 50 mL of DMF was added
and the solution was stirred for 16 hours. The red solution was diluted with 1
L of H20 and acidified to pH 3 with 3N HCI. The aqueous solution was
extracted with dichloromethane. The combined organics were washed with 3N
HCI, dried with Na2S04 and the solvents removed at reduced pressure to yield
crude 3,5-dichlorophenylacetonitrile which was used without purification.
The nitrile was added to a mixture of 40 mL of concentrated sulfuric
acid and 50 mL H20 and heated to reflux for 48 hours, cooled to room
temperature and stirred for 48 hours. The reaction was diluted into 1 L of
crushed ice, warmed to toom temperature and extracted with 2 x 200 mL of
dichloromethane and 2 x 200 mL of ethylacetate. Both sets of organics were
combined and washed with saturated aqueous NaHC03. The NaHC03 fractions
were combined and acidified to pH 1 with 3N HCI. The white solid was too
fine to filter and was extracted out with 2 X 200 mL of dichloromethane. The
combined organics were dried with Na~S04 and the solvents removed at reduced
presure to yield crude 3,5-dichlorophenylacetic acid as a white solid. The
solid
was slurried with hexane and filtered to get 1.75g of white solid.
NMR (CDC13): (in ppm) 3.61 (s, 2H), 7.19 (s,1H), 7.30 (s, 1H)
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Example BD
Synthesis of N-(3-chlorophenylacetyl)alanine
The title compound was prepared using L-alanine (Nova Biochem) and
3-chlorophenyl acetic acid (Aldrich) by following General Procedures BF or
' S BG, followed by hydrolysis using General Procedure BD.
Example B1
Synthesis of N-(phenylacetyl)-D, L-alanine iso-butyl ester
Following General Procedure BA above and using phenylacetyl chloride
(Aldrich) and D, L-alanine iso-butyl ester hydrochloride (from Example BB
above), the title compound was prepared. The reaction was monitored by tlc
on silica gel and purification was by extraction with Et~O followed by washes
with aqueous K~C03 and aqueous HCI.
NMR data was as follows:
'H-nmr (CDCI,): S = 7.23-7.36 (m, 5H), 6.18 (d, 1 H), 4.58 (t, J = 7.3
Hz, IH), 3.87 (m, 2H), 3.57 (s, 2H}, 1.90 (m, 1H), 1.34 (d, J= 7.2 Hz, 3H),
0.89 (d, J = 6.8 Hz, 6H).
'3C-nmr (CDCI,): 8 = 172.7, 170.3, 134.5, 129.2, 128.8, 127.2, 71.3,
48.1, 43.4, 27.5, 18.8, 18.3.
C,SH~,N03 (MW = 263.34; Mass Spectroscopy (MH' = 264))
Example B2
Synthesis of N (3-phenylpropionyl)-D,L-alanine iso-butyl ester
Following General Procedure BA above and using 3-phenylpropionyl
chloride (Aldrich) and D,L-alanine iso-butyl ester hydrochloride (from Example
BB above), the title compound was prepared as a solid having a melting point
of
_ from 51 °-54°C. The reaction was monitored by tlc on silica
gel and purification
was by extraction with Et,O followed by washes with aqueous K,C03 and
aqueous HCI.
NMR data was as follows:
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'H-nmr (CDCI,): 8 = 7.25 (m, 2H), 7.19 (m, 3H), 6.28 (d, J-= 7.2 Hz,
1 H), 4.58 (quint., J = 7.2 Hz, 1 H), 3.89 (m, 2H), 2.95 (t, J = 7.7 Hz, 2H),
2.50
(m, 2H), 1.92 (m, 1H), 1.33 (d, J = 7.1 Hz, 3H), 0.91 (d, J = 6.7 Hz, 6H).
'3C-nmr (CDCl3): 8 = 173.0, 171.5, 140.6, 128.3, 128.1: 126.0, 71.2,
47.8, 37.9, 31.4, 27.5, 18.79, 18.77, 18.3.
C,6H23NO3 (MW = 277.37, Mass Spectroscopy (MH+ 278))
Example B3
Synthesis of N (3-methylpentanoyl)-L-alanine iso-butyl ester
Following General Procedure BB and using 3-methylpentanoic acid
(Aldrich) and L-alanine iso-butyl ester hydrochloride (from Example BB above),
the title compound was prepared as an oil. The reaction was monitored by tIc
on silica gel and purification was by extraction with Et,O followed by washes
with aqueous K,C03 and aqueous HCI.
NMR data was as follows:
'H-nmr (CDCl3}: 8 = 6.08 (d, J = 5.9 Hz, 1H), 4.62 {quint., J = 7.3 Hz,
1 H), 3.92 (m, 2H), 2.22 (m, 1 H), 1.84-2.00 (m, 3H), 1.40 (d, J = 7.2 Hz,
3H),
1.35 (m, 1H), 1.20 (m, 1H), 0.85-0.96 (m, 12H).
'3C-nmr (CDC13): 8 = 173.3, 172.1, 71.4, 47.9, 43.9, 32.3, 29.38, 29.35,
27.6, 19.10, 19.06, 18.93, 18.91, 18.72, 18.67, 11.3.
C,3HZSN03 (MW = 243.35, Mass Spectroscopy (MH+ 244))
Example B4
Synthesis of N [(4-chlorophenyl)acetyl]-L-alanine iso-butyl ester
Following General Procedure BB and using 4-chlorophenylacetic acid
(Aldrich) and L-alanine iso-butyl ester hydrochloride (from Example BB above),
the title compound was prepared as a solid having a melting point of 11 I
°-
I 13 °C. The reaction was monitored by tlc on silica gel and
purification was by
extraction with Et,O followed by washes with aqueous K,C03 and aqueous HCI.
NMR data was as follows:
r
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'H-nmr (CDCI,): 8 = 7.30 (d, J = 8.2 Hz, 2H), 7.21 (d, J = 8.3 Hz, 2H),
6.18 (d, J = 5.5 Hz, 1 H), 4.57 (quint., J = 7.2 Hz, 1 H), 3.88 (m, 2H), 3.53
(s,
2H), I .91 {m, 1H), 1.36 (d, J = 7.1 Hz, 3H), 0.90 (d, J = 6.8 Hz, 6H).
'3C-nmr (CDCl3): 8 = 172.8, 169.8, 133.1, 133.0, 130.6,- 128.9, 71.4,
48.2, 42.6, 27.6, 18.85, 18.82, 18.4.
C,SH~oN03Cl (MW = 297.78, Mass Spectroscopy (MH+ 298))
Example BS
Synthesis of N ((3,4-dichlorophenyl)acetyl]-L-alanine iso-butyl ester
Following General Procedure BB and using 3,4-dichlorophenylacetic acid
(Aldrich) and L-alanine iso-butyl ester hydrochloride (from Example BB above))
the title compound was prepared as a solid having a melting point of 81
°-83 °C.
The reaction was monitored by tlc on silica gel and purification was by
extraction with Et20 followed by washes with aqueous KzC03 and aqueous HCI.
NMR data was as follows:
'H-nmr (CDCI,): 8 = 0.90 {d, J = 6.8 Hz, 6H), 1.38 (d. .1= 7.1 Hz, 3H),
1.91 (m, 1 H), 3.50 (s, 2H), 3.90 {m, 2H), 4.57 (quint., J = 7.1 Hz, 1 H),
6.31 (d,
J = 4.9 Hz, 1H),7.12 (m, 1H), 7.38 (m, 2H).
'3C-nmr (CDCl3): 8 = 18.4, 18.8, 18.9, 27.6, 42.2, 48.3, 71.5, 128.6,
130.6, 131.2, 131.3, 132.6, 134.7, 169.2, 172.8.
C,SH,9N03C1, (MW = 332.23, Mass Spectroscopy (MH+ 332))
Example B6
Synthesis of N [(4-methylphenyl)acetyl]-D,L-alanine iso-butyl ester
Following General Procedure BB and using 4-methylphenylacetic acid
(Aldrich) and D,L-alanine iso-butyl ester hydrochloride (from Example BB
above), the title compound was prepared as a solid having a melting point of
102°-104°C. The reaction was monitored by tlc on silica gel (Rf
=0.6 in 33%
ethyl acetate/hexanes) and purification was by extraction with Et~O followed
by
washes with aqueous K,C03 and aqueous HC1.
NMR data was as follows:
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'H-nmr (CDCI,): 8 = 0.90 (d, J = 6.7 Hz, 6H), 1.35 (d, J = 7.2 Hz, 3H),
1.91 (m, 1 H), 2.34 (s, 3H), 3.55 (s, 2H), 3.88 (m, 2H), 4.58 (m, 1 H), 6.05
(bd,
1 H), 7.16 (s; 4H).
'3C-nmr (CDCI3): 8 = 18.5, 18.85, 18.87, 21.0, 27.6, 43:1, 48.1, 71.3,
S 129.2, 129.6, 131.3, 136.9, 170.6, 172.8.
C~6H23NO3 (MW = 277.37, Mass Spectroscopy (MH' 278))
Example B7
Synthesis of N [(3-pyridyl)acetyl]-D,L-alanine iso-butyl ester
Following General Procedure BF and using 3-pyridylacetic acid
hydrochloride (Aldrich) and D,L-alanine iso-butyl ester hydrochloride (from
Example BB above), the title compound was prepared as a solid having a
melting point of 62°-64°C. The reaction was monitored by tlc on
silica gel (Rf
= 0.48 10% methanol/dichloromethane) and purification was by silica gel
chromatography.
NMR data was as follows:
'H-nmr (CDCI,): b = 8.40 (d, J= 2.8, 2H); 7.6 (m, 1H): 7.16 (m, 2H);
4.5 (quint., J = 7.2, 7.2, 1 H); 3.8 (m, 2H); 3.48 {s, 2H); 1.8 (m, 1 H); 1.30
(d, J
= 7.2, 3H); 0.81 (d, J = 6.7, 6H).
'3C-nmr (CDCl3): S = 173.4, 170.1, 150.6, 148.8, 137.4, 131.4, 124.1,
71.9, 48.9, 40.6, 28.1, 19.5, 19.4, 18.6.
C,QH~oN203 (MW = 264, Mass Spectroscopy (MH' 265))
Example B8
Synthesis of N [(1-naphthyl)acetyl]-L-alanine iso-butyl ester
Following General Procedure BB and using 1-naphthylacetic acid
(Aldrich) and L-alanine iso-butyl ester hydrochloride (from Example BB above),
the title compound was prepared as a solid having a melting point of
69°-73°C.
The reaction was monitored by tlc on silica gel and purification was by
extraction with Et~O followed by washes with aqueous K.,C03 and aqueous HCI.
NMR data was as follows:
.... _.........._._........,..._.. ,~.,.e.,._,_ ..,.... ~..... .. f..
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'H-nmr (CDCI,): 8 = 0.83 (m, 6H), 1.25 (d, J = 7.1 Hz, 3H); 1.81 (m,
1 H), 3.79 (m, 2H), 4.04 (2s, 2H), 4.57 (quint., J = 7.3 Hz, 1 H), 5.99 (d, J
= 7.1
Hz, I H), 7.44 (m, 2H), 7.53 (m, 2H), 7.85 (m, 2H), 7.98 {m, I H).
'3C-nmr (CDC13): 8 = 18.2, 18.81, 18.83, 27.5, 41.5, 48.2, 71.3, 123.7,
125.6, 126.1, 126.6, 128.2, 128.5, 128.7, 130.7, 132.0, 133.9, 170.3, 172.5.
C,9H,3NO3 (MW = 313.40, Mass Spectroscopy (MH+ 314))
Example B9
Synthesis of N [(2-naphthyl)acetyl]-L-alanine iso-butyl ester
Following General Procedure BB and using 2-naphthylacetic acid
(Aldrich) and L-alanine i.so-butyl ester hydrochloride (from Example BB
above),
the title compound was prepared as a solid having a melting point of
128°-
129°C. The reaction was monitored by tlc on silica gel and purification
was by
extraction with Et~O followed by washes with aqueous K~C03 and aqueous HCI.
NMR data was as follows:
'H-nmr (CDCI,): 8 = 0.86 (m, 6H), 1.35 (d, J = 7.1 Hz, 3H), 1.78 (m,
1 H), 3.76 (s, 2H), 3.87 (m, 2H), 4.62 (quint., J = 7.2 Hz, I I-1), 6.13 (d, J
= 7.1
Hz, 1H), 7.41 (m, 1H), 7.48 (m, 2H), 7.74 (s, IH), 7.83 (m, 3H).
'3C-nmr (CDC13): 8 = i8.4, 18.82, 18.85, 27.6, 43.7, 48.2, 71.4, 125.9,
126.3, 127.2, 127.6, 127.7, 128.2, 128.7, 132.0, 132.5, 133.5, 170.3, 172.8.
C,9H"N03 (MW = 313.40, Mass Spectroscopy (MH+ 3I4)).
Example B 10
Synthesis of N {4-phenyibutanoyl)-L-alanine iso-butyl ester
Following General Procedure BB and using 4-phenylbutanoic acid
(Aldrich) and L-alanine iso-butyl ester hydrochloride (from Example BB above),
the title compound was prepared as an oil. The reaction was monitored by tlc
on silica gel and purification was by extraction with Et,O followed by washes
with aqueous KZC03 and aqueous HCI.
NMR data was as follows:
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'H-nmr (CDCI,): b = 0.92 (d, J = 6.7 Hz, 6H), 1.38 (d, J = 7.1 Hz, 3H),
1.96 (m, 3H), 2.21 (t, J = 7.1 Hz, 2H), 2.64 (t, J = 7.3 Hz, 2H), 3.90 (m,
2H),
4.59 (quint.,-J = 7.2 Hz, 1 H), 6.31 (d, 1I-I), 7.16 (m, 3H), 7.24 (m, 2H).
'3C-nmr (CDCI3): 8 = 18.3, 18.75, 18.78, 26.8, 27.5, 34:9, 35.3, 47.8,
7I.2, 125.7, 128.2, 128.3, 141.3, 172.1, 173Ø
C"H,SN03 (MW = 291.39, Mass Spectroscopy (MH+ 292)).
Example B 11
Synthesis of N (5-phenylpentanoyl)-L-alanine iso-butyl ester
Following General Procedure BB and using 5-phenylpentanoic acid
(Aldrich) and L-alanine iso-butyl ester hydrochloride (from Example BB above),
the title compound was prepared as an oil. The reaction was monitored by tlc
on silica gel and purification was by extraction with Et,O followed by washes
with aqueous K,C03 and aqueous HCI.
NMR data was as follows:
'H-nmr (CDCI,): b = 7.23 (m, 2H), 7.17 (m, 3H), 6.30 (d, 1H), 4.59
(quint., J = 7.3 Hz, 1 H), 3.91 (m, 2H), 2.61 (t, J = 7.2 Hz, 2H), 2.22 (t, J
= 7.2
Hz, 2H), 1.93 (m, 1H), 1.66 (m, 4H), 1.38 (d, J = 7.2 Hz, 3H), 0.92 (d, J =
6.7
Hz, 6H).
''C-nmr (CDC13): ~ = 173.1, 172.3, 142.0, 128.2, 128.1, 125.6, 71.2,
47.8, 36.1, 35.5, 30.8, 27.5, 25.0, 18.80, 18.77, 18.4.
C,BHz.,N03 (MW = 305.39, Mass Spectroscopy (MH+ 306)).
Example B 12
Synthesis of N [(4-pyridyl)acetyl]-D,L-alanine iso-butyl ester
Following General Procedure BF and using 4-pyridylacetic acid
hydrochloride (Aldrich) and (D,L)-alanine iso-butyl ester hydrochloride (from
Example BB above), the title compound was prepared as a solid having a
melting point of 64°-66°C. The reaction was monitored by tlc on
silica gel (Rf
= 0.43 10% methanol/dichloromethane) and purification was by silica gel
chromatography.
_._._.__.~~.-,~.._.,..... . .~ >,~,..
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NMR data was as follows:
'H-nmr (CDCI,): 8 = 8.51 (dd, J = 1.6, 2.8, 1.6, 2H); 7.23 (dd, J = 4.3,
I .6, 4.4, 2H); 6.71 (d, J = 6.8, 1 H); 4.56 ( quint., J = 7.3, 7.2, I H);
3.88 (m,
2H); 3.53 (s, 2H); 1.89 (m, 1H); 1.36 (d, J = 7.2, 3H); 0.88 (d,-J = 6.7, 6H).
''C-nmr (CDC13): 8 = 173.5, 169.3, 150.5, 144.4, 125.1, 72.1, 48.9, 43.0,
28.2, 19.5, 19.5, 18.9.
C,~H,oN,O~ (MW = 264, Mass Spectroscopy (MH* 265))
Example B 13
Synthesis of N-(phenylacetyl)-L-alanine iso-butyl ester
Following General Procedure BB and using phenylacetyl chloride
{Aldrich) and L-alanine i.so-butyl ester hydrochloride (from Example BB
above),
the title compound was prepared as a solid having a melting point of
45°-47°C.
The reaction was monitored by tlc on silica gel and purification was by
extraction with Et~O followed by washes with aqueous K,C03 and aqueous HCi.
NMR data was as follows:
'H-nmr (CDCIj): 8 = 7.24-7.39 (m, SH), 6.14 (d, 1 H), 4.58 (t, J = 7.3
Hz, 1 H), 3.88 (m, 2H), 3.58 (s, 2H), 1.90 I;m, 1 H), 1.35 (d, J = 7.2 Hz,
3H),
0.89 (d, J = 6.7 Hz, 6H).
''C-nmr (CDCl3): 8 = 172.8, 170.4, 134.5, 129.3, 128.9, 127.2, 71.3,
48.1, 43.5, 27.5, 18.9, 18.8, 18.4.
C,SH,,N03 (MW = 263.34, Mass Spectroscopy (MH+ 264)).
Example B14
Synthesis of 2-[(3,4-dichlorophenyl)acetamido]butyric acid iso-butyl ester
Following General Procedure BI ab«ve and using 3,4-
dichlorophenylacetic acid (Aldrich) and iso-butyl 2-aminobutyrate (prepared
following General Procedure BJ above) the title compound was prepared. The
reaction was monitored by tlc on silica gel and purification was by filtration
as
described in the general procedure.
NMR data was as follows:
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'H-nmr (CDCI,): 8 = 7.36 (m, 3H), 6.03 (bd, 1H), 4.54 (m, 1H), 3.87
(m, 2H), 3.49 (s, 2H), 1.93 (m, 2H), I .72 (m, I H), 0.88 (d, 6H), 0.80 (t,
3H).
Example B15 -
Synthesis of 2-[(3-methoxyphenyl)acetamidoJbutyric acid iso-butyl ester
Following General Procedure BI above and using 3-methoxyphenylacetic
acid (Aldrich) and iso-butyl 2-aminobutyrate (prepared frollowing General
Procedure BJ above), the title compound was prepared. The reaction was
monitored by tlc on silica gel and purification was by filtration as described
in
the general procedure.
NMR data was as follows:
'H-nmr (CDCI,): 8 = 6.75 (m, 4H), 5.93 (bd, 1 H), 4.51 (m, 1 H), 3.83
(m, 2H), 3.75 {s, 2H), 3.52 (s, 2H), 1.82 (m, 2H), 1.60 (m, 1 H), 0.84 (d,
6H),
0.74 (t, 3H).
C,~H,5N04 (MW = 307.39, Mass Spectroscopy (MH+ 309)).
Example B 16
Synthesis of 2-[(4-nitrophenyl)acetamido]butyric acid iso-butyl ester
Following General Procedure BI above and using 4-nitrophenylacetic
acid (Aldrich} and iso-butyl 2-aminobutyrate {prepared following General
Procedure BJ above), the title compound was prepared. The reaction was
monitored by tlc on silica gel and purification was by filtration as described
in
the general procedure.
NMR data was as follows:
'H-nmr (CDCI,): 8 = 8.16 (d, 2H), 7.44 (d, 2H), 6.04 (bd, 1 H), 4.55 (m,
1 H), 3.86 (m, 2H), 3.66 (s, 2H), 1.86 (m, 2H), 1.67 (m, 1 H), 0.85 (d, 6H),
0.81
(t, 3H).
C,6H~~N~05 (MW = 322.36, Mass Spectroscopy (MH+ 323)).
Example B17
Synthesis of 2-[(3,4-methylenedioxyphenyl)acetamido]butyric acid
' iso-butyl ester
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Following General Procedure BI above and using 3,4-{methylenedioxy)-
phenyl acetic acid (Aldrich) and iso-butyl 2-aminobutyrate (prepared following
General Procedure BJ above), the title compound was prepared. The reaction
was monitored by tlc on silica gel and purification was by filtration as
described
in the general procedure.
NMR data was as follows:
'H-nmr (CDCI,): 8 = 6.72 (m, 3H), 5.92 (bd, IH), 4.54 (m, 1H), 3.86
(m, 2H), 3.66 {s, 2H), 1.86 (m, 2H), 1.66 (m, 1 H), 0.89 (d, 6H), 0.79 (t,
3H).
Example B I 8
Synthesis of 2-[(thien-3-yl)acetamido]butyric acid iso-butyl ester
Following General Procedure BI above and using 3-thiopheneacetic acid
(Aldrich) and iso-butyl 2-aminobutyrate (prepared following General Procedure
BJ above), the title compound was prepared. The reaction was monitored by tlc
on silica gel and purification was by filtration as described in the general
procedure.
NMR data was as follows:
'H-nmr (CDCI,): 8 = 7.37 (m, 1H), 7.16 (m, IH), 7.04 (m, 1H), 6.05
(bd, 1 H), 4.57 (m, 1 H), 3.66 (s, 2H), 1.93 (m, 2H), 1.67 (m, 1 H), 0.91 (d,
6H),
0.86 (t, 3H).
Example B 19
Synthesis of 2-[(4-chlorophenyl)acetamido]butyric acid iso-butyl ester
Following General Procedure BI above and using 4-chlorophenylacetic
acid (Aldrich) and iso-butyl 2-aminobutyrate (prepared following General
Procedure BJ above), the title compound was prepared. The reaction was
monitored by tlc on silica gel and purification was by filtration as described
in
the general procedure.
NMR data was as follows:
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'H-nmr (CDCI;): 8 = 7.22 (m, 2H), 7.I1 (m, 2H), 5.80 (m, 1-H), 4:44 (m,
1 H), 3.78 (m, 2H), 3.43 (s, 2H), 1.77 (m, 2H), 1.56 (m, 1 H), 0.83 (d, 6H)
0.71
(t, 3H). ' -
Example B20
Synthesis of 2-((3-nitrophenyl)acetamido] butyric acid iso-butyl ester
Following General Procedure BI above and using 3-nitrophenylacetic
acid (Aldrich) and iso-butyl 2-aminobutyrate (prepared following General
Procedure BJ above), the title compound was prepared. The reaction was
monitored by tlc on silica gel and purification was by filtration as described
in
the general procedure.
NMR data was as follows:
'H-nmr (CDC13): 8 = 8.15 (m, 2H), 7.65 (m, 1H), 6.08 (m, 1H), 4.46 (m,
1 H), 3.92 (m, 2H), 3.68 (s, 2H), 1.91 (m, 2H), 1.75 (m, 1 H), 0.98 {d, 6H)
0.71
(t, 3H).
Example B21
Synthesis of 2-[(2-hydroxyphenyl)acetamido] butyric acid iso-butyl ester
Following General Procedure BI above and using 2-hydroxyphenylacetic
acid (Aldrich) and iso-butyl 2-aminobutyrate (prepared following General
Procedure BJ above), the title compound was prepared. The reaction was
monitored by tlc on silica gel and purification was by filtration as described
in
the general procedure.
NMR data was as follows:
'H-nmr (CDCI,): 8 = 7.14 (m, 1 H), 7.01 (m, 1 H), 6.93 (m, 1 H), 6.79 (m,
1 H), 6.46 (m, 1 H), 4.51 (m, 1 H), 3.87 (m, 2H), 3.57 (s, 2H), 2.01 (m, 2H),
1.75
(m, 1H), 0.89 (d, 6H), 0.85 (t, 3H).
Example B22
Synthesis of 2-[(2-naphthyl)acetamido]butyric acid iso-butyl ester
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Following General Procedure BI above and using 2-naphthylacetic acid
- (Aldrich) and iso-butyl 2-aminobutyrate (prepared following General
Procedure
' BJ above), the title compound was prepared. The reaction was monitored by
tlc
on silica gel and purification was by filtration as described in the general
' S procedure.
NMR data was as follows:
'H-nmr (CDCI,): ~ = 7.83 (m, 7H), 5.95 (m, I H), 4.58 {m, 1 H), 3.84 (m,
2H), 3.75 (s, 2H), 1.89 (m, 2H), 1.63 (m, 1H), 0.91 (d, 6H), 0.81 (t, 3H).
C~oH~5N0~ (MW = 327.42, Mass Spectroscopy (MH' 328)).
Example B23
Synthesis of 2-[(2,4-dichlorophenyl)acetamido] butyric acid iso-butyl ester
Following General Procedure BI above and using 2,4-
dichlorophenylacetic acid (Aldrich) and iso-butyl 2-aminobutyrate (prepared
following General Procedure BJ above), the title compound was prepared. The
reaction was monitored by tlc on silica gel and purification was by filtration
as
described in the general procedure.
NMR data was as follows:
~H-nmr {CDCI;): 8 = 7.49 (m, I H). 7.22 (m, 2H) 5.98 (m, 1 H), 4.52 (m,
1 H), 3.86 (m, 2H), 3.61 (s, 2H), 1.84 (m, 2H), 1.62 (m, 1 H) 0.87 (d, 6H),
0.80
(t, 3H).
Example B24
Synthesis of 2-[(4-bromophenyl)acetamido]butyric acid iso-butyl ester
Following General Procedure BI above and using 4-bromophenylacetic
acid (Aldrich) and iso-butyl 2-aminobutyrate (prepared following General
Procedure BJ above), the title compound was prepared. The reaction was
monitored by tlc on silica gel and purification was by filtration as described
in
the general procedure.
NMR data was as follows:
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'H-nmr (CDCI,): 8 = 7.43 (d, 2H), 7.19 (d, 2H) 5.85 (m, I H-), 4.5-1 (m,
1 H), 3.81 (m, 2H), 3.47 (s, 2H), 1.84 (m, 2H), 1.61 (m, 1 H) 0.84 {d, 6H),
0.76
(t~ 3H). - _
C~6H,~N03Br (MW = 356.26, Mass Spectroscopy (MH+ -358)).
Example B25
Synthesis of 2-[(3-chlorophenyl)acetamido])butyric acid iso-butyl ester
Following General Procedure BI above and using 3-chlorophenylacetic
acid (Aldrich) and iso-butyl 2-aminobutyrate (prepared following General
Procedure BJ above), the title compound was prepared. The reaction was
monitored by tlc on silica gel and purification was by filtration as described
in
the general procedure.
NMR data was as follows:
'H-nmr (CDCI,): 8 = 7.25 (m, 3H), 7.12 (m, 1 H) 5.80 (m, 1 H), 4.52 (m,
I H), 3.86 (m, 2H), 3.50 (s, 2H), 1.87 (m, 2H), 1.67 (m, 1 H) 0.88 (d, 6H),
0.77
(t, 3H}.
C,6HZ~N03C1 (MW = 311.81 Mass Spectroscopy (MH~ 313)).
Example B26
Synthesis of 2-[(3-fluorophenyl)acetamido]butyric acid iso-butyl ester
Following General Procedure BI above and using 3-fluorophenylacetic
acid (Aldrich) and iso-butyl 2-aminobutyrate {prepared following General
Procedure BJ above}, the title compound was prepared. The reaction was
monitored by tlc on silica gel and purification was by filtration as described
in
the general procedure.
NMR data was as follows:
'H-nmr (CDCI,): 8 = 7.31 (m, IH), 7.01 {m, 3H) 5.95 (rn, 1H), 4.54 (m,
1H), 3.84 (m, 2H), 3.54 (s, 2H), 1.88 (m, 2H), 1.65 (m, 1H) 0.87 (d, 6H), 0.81
(t, 3H).
C,6H"N03F (MW = 295.35 Mass Spectroscopy ~(MH+ 296)).
. . ...~... .......... . ,.. .
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Example B27
Synthesis of 2-[(benzothiazol-4-yl)acetamido]butyric acid iso-butyl ester
Following General Procedure BI above and using 4-benzothiazol-4-yl
acetic acid (Chemservice) and iso-butyl 2-aminobutyrate (prepared following
General Procedure BJ above), the title compound was prepared. The reaction
was monitored by tic on silica gel and purification was by filtration as
described
in the general procedure.
NMR data was as follows:
'H-nmr (CDCI,): 8 = 7.82 (m, 1H), 7.51-7.21 (m, 4H) 5.84 (m, 1H), 4.51
(m, 1 H), 3.90 (s, 2H), 3.79 (m, 2H), 1.78 (m, 2H), 1.58 {m, 1 H) 0.80 (d,
6H),
0.66 (t, 3H).
Example B28
Synthesis of 2-[(2-methylphenyl)acetamido] butyric acid iso-butyl ester
Following General Procedure BI above and using 2-methylphenylacetic
acid (Aldrich) and iso-butyl 2-aminobutyrate {prepared following General
Procedure BJ above), the title compound was,prepared. 'The reaction was
monitored by tlc on silica gel and purification was by filtration as described
in
the general procedure.
NMR data was as follows:
'H-nmr {CDCI,): b = 7.18 (m, 4H), 5.79 (m, 1 H), 4.54 (m, i H), 3.85 (m,
2H), 3.59 (s, 2H), 3.29 (s, 3H), 1.81 (m, 2H), 1.59 (m, 1 H) 0.87 (d, 6H),
0.77
(t, 3H).
C"HZSN03 (MW = 291.39 Mass Spectroscopy (M+ 291)).
Example B29
Synthesis of 2-[(2-fluorophenyl)acetamido]butyric acid iso-butyl ester
Following General Procedure BI above and using 2-fluorophenylacetic
acid (Aldrich) and iso-butyl 2-aminobutyrate (prepared following General
Procedure BJ above), the title compound was prepared. The reaction was
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monitored by tlc on silica gel and purification was by filtration as described
in
the general procedure.
NMR data was as follows:
'H-nmr (CDCI,): 8 = 7.28 (m, 1H), 7.09 (m, 3H) 6.03 (m, 1H), 4.54 (m,
1 H), 3.87 (m, 2H), 3.57 (s, 2H), 1.89 (m, 2H), 1.64 (m, 1 H) 0.88 (d, 6H),
0.80
(t, 3H).
Example B30
Synthesis of 2-[(4-fiuorophenyl)acetamido]butyric acid iso-butyl ester
Following General Procedure BI above and using 4-fluorophenylacetic
acid {Aldrich) and iso-butyl 2-aminobutyrate (prepared following General
Procedure BJ above), the title compound was prepared. The reaction was
monitored by tlc on silica gel and purification was by filtration as described
in
the general procedure.
NMR data was as follows:
'H-nmr (CDCI,): b = 7.20 (m, 2H), 6.97 (m, 2H) 5.87 (m, 1H), 4.492
(m, 1H), 3.83 (m, 2H), 3.48 (s, 2H), 1.86 (m, 2H), 1.60 (m, 1 H) 0.87 (d, 6H),
0.78 (t, 3H).
C,6H,~N03F (MW = 295.35 Mass Spectroscopy (MH+ 296)).
Example B31
Synthesis of 2-[(3-bromophenyl)acetamido)butyric acid iso-butyl ester
Following General Procedure BI above and using 3-bromophenylacetic
acid (Aldrich) and iso-butyl 2-aminobutyrate (prepared following General
Procedure BJ above), the title compound was prepared. The reaction was
monitored by tlc on silica gel and purification was by filtration as described
in
the general procedure.
NMR data was as follows:
'H-nmr (CDC1,): 8 = 7.45 (m, 2H), 7.23 (m, 2H) 5.95 (m, 1H), 4.55 (m,
1H) 3.84 (m, 2H) 3.55 (s, 2H), 1.89 (m, 2H), 1.68 (m, 1H) 0.91 (d, 6H), 0.81
(t, 3H).
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C,6HZ,N03Br (MW = 356.26 Mass Spectroscopy (M+ 357)). -
- Example B32
Synthesis of 2-[(3-trifluoromethylphenyl)acetamido]butyric acid
iso-butyl ester
Following General Procedure BI above and using 3-trifluoromethyl-
phenylacetic acid (Aldrich) and i.so-butyl 2-aminobutyrate (prepared following
General Procedure BJ above), the title compound was prepared. The reaction
was monitored by tlc on silica gel and purification was by filtration as
described
in the general procedure.
NMR data was as follows:
' H-nmr (CDCI,): 8 = 7.52 (m, 1 H), 7.47 (m, 2H) 6.01 (m, 1 H), 4.56 (m,
1 H), 3.86 (m, 2H), 3.61 (s, 2H), 1.84 (m, 2H), 1.62 (m, 1 I-I) 0.87 (d, 6I-
I), 0.80
(t, 3H}.
C,~H"N03F3 (MW = 345.36 Mass Spectroscopy (MH' 345)).
Example B33
Synthesis of 2-[(2-thienyl)acetamidoJbutyric acid iso-butyl ester
Following General Procedure BI above and using 2-thiopheneacetic acid
(Aldrich) and iso-butyl 2-aminobutyrate (prepared following General Procedure
BJ above), the title compound was prepared. The reaction was monitored by tlc
on silica gel and purification was by filtration as described in the general
procedure.
NMR data was as follows:
'H-nmr (CDCI,): ~ = 6.89 (m, 3H), 6.07 (bd, 1H), 4.50 (m, 1H), 3.82
(m, 2H), 3.71 (s, 2H), 1.85 (m, 2H), 1.62 (m, 1 H), 0.81 (d, 6H), 0.75 (t,
3H).
C,4HZ,N03S (MW = 283.39, Mass Spectroscopy (MH' 284}).
Example B34
Synthesis of 2-(phenylacetamido)butyric acid iso-butyl ester
Following General Procedure BH above and using phenylacetic acid
(Aldrich) and iso-butyl 2-aminobutyrate (prepared following General Procedure
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BJ above), the title compound was prepared. The reaction was monitored by tlc
on silica gel and purification was by chromatography on silica gel using 9:1
toluene:EtOAc as the eluant.
NMR data was as follows:
'H-nmr (CDCI,): 8 = 7.17-7.28 (m, SH), 6.23 (bd, 1H), 4.51 (m, 1H),
3.86 (m, 2H), 3.54 (s, 2H}, 1.87 {m, 2H), i .62 (m, 1 H), 0.87 (d, 6H), 0.78
(t,
3 H).
C,6Hz3NO3 (MW = 277.36, Mass Spectroscopy (MH+ 277)).
Example B35
Synthesis of N (phenylacetyl)valine 2-methylbutyl ester
Step A. Preparation of N-(phenylacetyl) valine
To a stirred solution of 5.15 g (44 mmol) of valine (Bachem) in 50 mL
( 100 mmol) of 2N NaOH cooled to 0°C was added dropwise 5.3 mL (40
mmol)
of phenylacetyl chloride (Aldrich). A colorless oil precipitated. The reaction
mixture was allowed to warm to room temperature and stirred for 18 hours,
washed with 50 mL diethyl ether, acidified to pH 2-3 with aqueous HCI. The
white precipitate formed was filtered off, washed thoroughly with water,
followed by diethyl ether to give 7.1 g (30 mmol, 69% yield) of the title
compound.
NMR data was as follows:
'H-nmr (DMSO-db): 8 = 12.63 (s, 1H), 8.25 (d, J = 8.6 Hz, 1H), 7.27
(m, SH), 4.15 (m, 1H), 3.56 (d, J = 13.8 Hz, 1H), 3.47 (d, J = 13.8 Hz, 1H},
2.05 (m, 1H), 0.87 (d, J = 6.8, Hz, 3H), 0.84 {d, J = 6.8 Hz, 3)
'3C-nmr (DMSO-db): 8 = 173.2, 170.4, 136.6, 129.0, 128.2, 126.3, 57.1,
41.9, 30.0, 19.2, 18.0
C,3H,.,N03 (MW=235.29; Mass Spectroscopy (MH+ = 236))
Step B. Synthesis of N (phenylacetyl)valine 2-methylbutyl ester
Following General Procedure BC and using the N-{phenylacetyl) valine
prepared in Step A above and 2-methylbutan-1-of (Aldrich), the title compound
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was prepared as a diastereomeric mixture. The reaction was monitored by tlc on
silica gel and purification was by filtration as described in the general
procedure.
NMK data was as follows:
~H-nmr (CDC13): 8 = 7.25-7.40 (m, SH), 5.95 (d, 1 H), 4.56 (m, 1 H),
' S 3.84-4.00 (m, 2H), 3.61 (s, 2H), 2.10 (m, 1 H), 1.68 (m, 1 H), 1.38 (m, 1
H), I .15
(m 1H), 0.82-0.94 (m, 9H), 0.76 (d, 3H).
"C-nmr (CDCI~): b = 171.84, 171.81, 170.7, 134.6, 129.31, 129.27,
128.9, 127.3, 69.8, 57.0, 43.7, 33.9, 31.3, 25.9, 25.8" 18.9, 17.4, 16.34)
16.27,
11.12, 11.07.
C,BH~,NO; (MW = 305.42, Mass Spectroscopy (MH 306)).
Example B36
Synthesis of N (phenylacetyl)-L-methionine iso-butyl ester
L-Methionine (0.129g, 0.869 mmols) (Aldrich) was taken-up in dioxane
(5.0 mL) and treated with a saturated solution of sodium bicarbonate (5.0 mL)
followed by phenylacetyl chloride {Aldrich) (O.I 14 mL, 0.822 mmols). After
stirring for 17 hours at room temperature the mixture was diluted with ethyl
acetate, the layers separated and the aqueous layer acidified to pH 2 with SN
HCI. The crude product was extracted into ethyl acetate, dried over sodium
sulfate, vacuum dried and used without further purification.
N-Phenylacetyl-L-methionine (0.1285 g, 0.447 mmol) was dissolved in
3.0 mL dioxane and iso-butyl alcohol (0.2 mL) and treated with EDC (0.094 g,
0.492 mmol), and catalytic DMAP (O.OlSg). After stirring for 17 hours at
23°C, the mixture was evaporated at reduced pressure to an oil, the
residue was
diluted in EtOAc and washed with 0.1 N HCl and saturated sodium bicarbonate.
Chromatography on silica gel using 98:2 C',HCI3IMeOH as eluant provided the
pure product.
NMR data was as follows:
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'H-nmr (CDCl3): 8 = 7.4-7.23 (m, SH); 6.14 (bd, 1 H), 4.70 (m, 1 H),
3.89 (d, 2H), 3.62 (s, 2H), 2.43 (m, 2H), 2.12 (m, I H), 1.93 (m, 2H), 0.94
(d,
6H). '
C"H,SN03S (MW = 323.17, Mass Spectroscopy (M+ 323)
Example B37
Synthesis of N (phenylacetyl)-L-leucine iso-butyl ester
L-Leucine {Aldrich) (0.114g, 0.869 mmols) was taken-up in dioxane (5.0
mL) and treated with a saturated solution of sodium bicarbonate (5.0 mL}
followed by phenylacetyl chloride (Aldrich) (0.114 mL, 0.822 mmols). After
stirring for 17 hours at room temperature the mixture was diluted with ethyl
acetate, the layers separated and the aqueous layer acidified to pH 2 with SN
HC1. The crude product was extracted into ethyl acetate, dried over sodium
sulfate, vacuum dried and used without further purification.
N-Phenylacetyl-L-leucine (0.0081 g, 0.038 mmol) was dissolved in 2.0
mL CHCI3 (EtOH free) and iso-butyl alcohol (0.055 mL) and treated with P-
EPC { 100 mg, 0.87 milliequivalents). The mixture was rotated for 4 days,
filtered through a plug of cotton and the filtrate evaporated at reduced
pressure
to an oil which was sufficiently pure for testing.
NMR data was as follows:
'H-nmr (CDCIz): S = 7.22 (m, SH), 5.57 (d, 1H), 4.35 (m, 1H), 3.35 (m,
3H), 1.35 (m, 4H), 0.68 (m, 9H).
C,BHz~N03 (MW = 305.40, Mass Spectroscopy (MT 305)).
Example B38
Synthesis of N [(3-chlorophenyl)acetyl]alanine 3-methylbut-2-enyl ester
Following General Procedure BC above and using N-(3-
chlorophenylacetyl alanine (from Example BD above) and 3-methylbut-2-en-1-of
(Aldrich), the title compound can be prepared. The reaction was monitored by
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tlc on silica gel and purification was by liquid chromatography using 30%
EtOAc/hexane as the eluant.
NMR data was as follows:
'H-nmr (CDCI,): S = 7.39-7.16 (m, 4H), 6.06 (bd, 1 H), 5.38-5.29 (m,
1 H), 4.63 (d, J = 9Hz, 2H), 3.56 (s, 2H), 1.79 (s, 3H), 1.7 (s, 3H), 1.39 (d,
J =
9Hz, 3H).
Example B39
Synthesis of N ((3-chlorophenyI)acetyl]alanine cyclopropylmethyl ester
Following General Procedure BC above, and using N-(3-
chlorophenylacetyl alanine {from Example BD above) and cyclopropylmethanol
(Aldrich), the title compound can be prepared. The reaction was monitored by
tlc on silica gel and purification was by liquid chromatography using 3:7
EtOAc:hexane as the eluant.
NMR data was as follows:
'H-nmr (CDCI,): 8 = 7.2-7.1 (m, 4H), 6.09 (bs, 1H), 4.6 (dq, J= 9 Hz,
1H), 3.96 (dd, J = 9Hz, 2H), 3.59 (s, 2H), 1.2 (d, J = 9Hz, 3H), 1.2-1.0 (m,
1H), 0.603-0.503 (m, 2H), 0.300-0.203 (m, 2H).
Example B40
Synthesis of N ((3-chlorophenyl)acetyl]alanine 2-thienylmethyl ester
Following General Procedure BC above, and using N-(3-
chlorophenylacetyl alanine (from Example BD above) and 2-thiaphenemethanol
(Aldrich) the title compound can be prepared. The reaction was monitored by
tlc on silica gel and purification was by liquid chromatography using 3:7
EtOAc:hexane as the eluant.
NMR data was as follows:
'H-nmr (CDC1,): 8 = 7.37-6.97 (m, 7H), 5.97 (q, J = 14 Hz, 2H), 4.6
(dq, .I = 9 Hz, 1H), 3.76 (s, 2H), 1.38 (d, J = 9Hz, 3H).
Example B41
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Synthesis of N [(3-chlorophenyl)acetyl]alanine
(1-methylcyclopropyl)methyl ester
Following General Procedure BC above, and using N-(3-
chlorophenylacetyl alanine {from Example BD above) and (I-
methylcyclopropyl)methanol (Aldrich) the title compound can be prepared. The
reaction was monitored by tlc on silica gel and purification was by liquid
chromatography using 3:7 EtOAc:hexane as the eluant.
NMR data was as follows:
'H-nmr (CDCI,): 8 = 8.6 (bd, J = 9 Hz, 1 H), 3.86 (q, J = 14 Hz, 2H),
3.4 (s, 2H), 2.29 (q, J = 9 Hz, 1 H), I .3 (d, J = 9Hz, 3H), 1.03 (s, 3H), 0.5-
0.4
(m, 2H), 0.4-0.28 (m, 2H).
Example B42
Synthesis of N [(3-chlorophenyl)acetyl]alanine 3-thienylmethyl ester
Following General Procedure BC above, and using N-(3-
chlorophenylacetyl alanine (from Example BD above} and 3-thiophenemethanol
(Aldrich) the title compound can be prepared. The reaction was monitored by
tlc on silica gel and purification was by liquid chromatography using 3:7
EtOAc:hexane as the eluant.
NMR data was as follows:
'H-nmr (CDCI,): 8 = 8.03 (bd, J= 9 Hz, 1H), 7.56-7.5 (m, 1H), 7.47
(bs, I H), 7.4-7.17 (m, 4H), 7.06 (d, J = 9 Hz, 1 H), 5.1 ( s, 2H), 4.3 (dq,
11-1),
1.3 (d, J = 9 Hz, 3H).
Example B43
Synthesis of N [(3-chlorophenyl)acetylJalanine 2-methylcyclopentyl ester
Following General Procedure BC above, and using N-(3-
chlorophenylacetyl alanine (from Example BD above) and 2-
methylcyclopentanol (Aldrich) the title compound can be prepared. The reaction
was monitored by tlc on silica gel and purification was by liquid
chromatography using 3:7 EtOAc:hexane as the eluant.
NMR data was as follows:
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'H-nmr (CDCI,): S = 7.39-7.16 (m, 4H), 6.3 (bd, I H}, 4.79-4.7 (m, 1 H),
4.6-4.25 (m, J = 9 Hz, 1 H), 3.577 (s, 2H), 2.09-1.8 (m, 2H), I .74-1.6 (m,
2H),
1.39 {dd, J = 9 Hz, 3H), 1.2 (dt, J = 9 Hz, 1 H), 0.979 (dd, J = 9 Hz, 2H)
C"H"N03C1 (MW = 323.82, Mass Spectroscopy (MH+ 323).
' S
Example B44
Synthesis of N [(3-chlorophenyl)acetyljalanine 2-methylprop-2-enyl ester
Following General Procedure BC above, and using N-(3-
chlorophenylacetyl alanine (from Example BD above) and 2-methylprop-2-en-1-
0l (Aldrich) the title compound can be prepared. The reaction was monitored by
tlc on silica gel and purification was by liquid chromatography using 3:7
EtOAc:hexane as the eluant.
NMR data was as follows:
'H-nmr (CDCI,): 8 = 7.39-7.16 (m, 4H), 6.03 (bs, IH}, 4.77 (s, 2H), 4.7-
4.29 (m, 3H), 2.59 (s, 2H), 1.73 {s, 3H), 1.43 {d, J = 9 Hz, 3H)
C,SH,gN03Cl (MW = 295.76, Mass Spectroscopy (MH+ 295)).
Example B45
Synthesis of N [(3-chlorophenyl)acetyl] alanine cyclohex-2-enyl ester
Following General Procedure BC above, and using N-(3-
chlorophenylacetyl alanine (from Example BD above) and cyclohex-2-en-1-of
(Aldrich) the title compound can be prepared. The reaction was monitored by
tlc on silica gel and purification was by liquid chromatography using 3:7
EtOAc:hexane as the eluant.
NMR data was as follows:
'H-nmr (CDCI,): 8 = 8.6 (bd, J = 9 Hz, 1 H), 7.4-7.2 (m, 4H), 6.0-5.8
(m, 1 H), 5.7-S.5 (m, 1 H), 5.1 (bs, 1 H), 4. I 3-4.29 (m, 1 H), 3.5 (s, 2H),
2.1-1.9
(m, 2H), 1.8-1.69 (m, 1H), 1.69-1.49 (m, 4H), 1.3 (dd, J= 9 Hz, 3H)
C,~HzoN03Cl (MW = 321.8, Mass Spectroscopy (MH' 321.2)).
Example B46
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Synthesis of N [(2-phenylbenzoxazol-5-yl)acetyl]alanine iso-butyl ester
Following General Procedure BI above, and using 5-(2-
phenylbenzoxazol)-yl-acetic acid (CAS# 62143-69-5) and alanine iso-butyl ester
(prepared following General Procedure BJ above), the title compound was
prepared.
NMR data was as follows:
'H-nmr (CDCI,): 8 = 8.24 (m, 3H), 7.68 (m, 1H), 7.51 {m, SH), 6.04 (m,
1 H), 4. 5 8 (m, 1 H), 3 . 85 (m, 2H), 3 .68 (s, 2H), 1.9 (m, 1 H), 1.3 5 {d,
3 H), 0. 87
(d. 6H).
C,~H,4N,04 (MW = 380, Mass Spectroscopy (MH+ 381 )).
Example B47
Synthesis of N-[(3-methylthiophenyl)acetyl] alanine iso-butyl ester
Following General Procedure BI above, and using 3-
methylthiophenylacetic acid (CAS# 18698-73-2) and alanine iso-butyl ester
(prepared following General Procedure BJ above), the title compound was
prepared. The reaction was monitored by tlc on silica gel and purification was
by filtration as described in the general procedure.
NMR data was as follows:
'H-nmr (CDCI,}: 8 = 7.14 {m, 2H), 7.01 (m, IH), 4.56 (m, 1H), 3.88 (m,
2H), 3.54 (s, 2H), 2.46 (s, 3H), 1.89 (m, 1H), 1.35 (d, 3H) 0.85 (d, 6H}.
C,6H,3N03S (MW = 309, Mass Spectroscopy {MH+ 310)).
Example B48
Synthesis of N 4-[(2-furyl)acetyl]alanine iso-butyl ester
Following General Procedure BI above, and using 2-furylacetic acid
(CAS# 2745-26-8) and alanine iso-butyl ester (prepared following General
Procedure BJ above), the title compound was prepared. The reaction was
monitored by tlc on silica gel and purification was by filtration as described
in
the general procedure.
NMR data was as follows:
. ..-,...~..,..._ ._....... . . T
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'H-nmr (CDCI,): 8 = 7.36 (m, 1 H), 6.34 (m, 1 H), 6.21 (m, 1 H), 4.56 (m,
1H), 3.91 (m, 2H), 3.61 (s, 2H), 1.92 (m, 1H), 1.38 (d, 3H) 0.89 (d, 6H).
C,3H;9N04 (MW = 253, Mass Spectroscopy {MH' 254)).
S Example B49
Synthesis of N [(benzofuran-2-yl)acetyl]alanine iso-butyl ester
Following General Procedure BI above, and using benzofuran-2-ylacetic
acid (Maybridge) and alanine iso-butyl ester (prepared following General
Procedure BJ above), the title compound was prepared. The reaction was
monitored by tlc on silica gel and purification was by filtration as described
in
the general procedure.
NMR data was as follows:
'H-nmr (CDCI,): 8 = 7.51 (m, 1 H), 7.44 (m, 1 H),7.25 (m, 2H), 6.67 (s,
1 H), 4.60 (m, 1 H), 3.87 (m, 2H), 3.77 (s, 2H), 1.88 (m, 1 H), 1.38 (d, 3H),
0.87
(d, 6H).
C"H,,N04 (MW = 303, Mass Spectroscopy (MH' 304)).
Example B50
Synthesis of N [(benzothiophen-3-yl)acetyl]alanine iso-butyl ester
Following General Procedure BI above, and using thianaphthen-3-ylacetic
acid (Lancaster) and alanine iso-butyl ester (prepared following General
Procedure BJ above)) the title compound was prepared. The reaction was
monitored by tlc on silica gel and purification was by filtration as described
in
the general procedure.
NMR data was as follows:
'H-nmr (CDCI,): 8 = 7.89 (m, 1H), 7.76 (m, 1H), 7.38 (m, 3H), 6.07 (m,
1 H), 4.57 (m, 1 H), 3.92 (m, 2H), 3.82 (s, 4H), 1.84 (m, 1 H), 1.32 (d, 3H)
0.85
(d, 6H).
C,.,H,,N03S (MW = 319, Mass Spectroscopy (MH+ 320)).
Example B51
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Synthesis of N [(2-chloro-5-thienyl)acetyt]alanine iso-buty-1 ester
Following General Procedure BI above, and using 5-chloro-2-
thienyl)aceti'c acid (CAS# 13669-19-7) and alanine iso-butyl ester (prepared
following General Procedure BJ above), the title compound was prepared. The
reaction was monitored by tlc on silica gel and purification was by filtration
as
described in the general procedure.
NMR data was as follows:
'H-nmr (CDCI,): 8 = 6.77 (m, 1 H), 6.68 (d, 1 H), 6.31 (bm, 1 H), 4.59
(m, 1H), 3.91 (m, 2H), 3.38 (s, 2H), 1.90 (m, 1H), 1.39 (d, 3H) 0.89 (d, 6H).
C,3H~8N03SCI (MW = 303, Mass Spectroscopy (MH+ 303)).
Example B52
Synthesis of N [(3-methylisoxazol-5-yl)acetyl]alanine iso-butyl ester
Following General Procedure BI above, and using (3-methyl-isoxazol-S-
yl)acetic acid (CAS# 19668-85-0) and alanine iso-butyl ester (prepared
following
General Procedure BJ above), the title compound was prepared. The reaction
was monitored by tlc on silica gel and purification was by f Itration as
described
in the general procedure.
NMR data was as follows:
'H-nmr (CDCI,): 8 = 6.07 (s, 2H), 4.56 (m, 1H), 3.92 (m, 2H), 3.68 (s,
2H), 2.29 (s, 3H), 1.94 (m, 1 H), 1.89 (d, 3H) 0.91 (d, 6H).
C,3H~oNz04 (MW = 268, Mass Spectroscopy (MH+ 269)).
Example B53
Synthesis of N [(2-phenylthiothienyl)acetyl]alanine iso-butyl ester
Following General Procedure BI above, and using (2-phenyl-
thiothienyl)acetic acid and alanine iso-butyl ester (prepared following
General
Procedure BJ above), the title compound was prepared. The reaction was
monitored by tlc on silica gel and purification was by filtration as described
in
the general procedure.
NMR data was as follows:
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' H-nmr (CDCI,): 8 = 7.2I -7.11 (m, 6H), 6.92 (d, 1 H), 4.56(m, 1 H), 3.87
(m, 2H), 3.72 (s, 2H), 1.94 (m, 1 H), 1.38 (d, 3H) 0.89 (d, 6H).
C,9H23NO3S, (MW = 377, Mass Spectroscopy (MH' 378)).
Example B54
Synthesis of N [(6-methoxybenzothiophen-2-yl)acetylJ alanine iso-butyl ester
Following General Procedure BI above, and using (6-
methoxythianaphthen-2-yl)acetic acid and alanine iso-butyl ester (prepared
following General Procedure BJ above), the title compound was prepared. The
reaction was monitored by tlc on silica gel and purification was by filtration
as
described in the general procedure.
NMR data was as follows:
'H-nmr (CDCI,): 8 = 7.59 (d, 1H), 7.33 (d, 1H), 7.16 (s, 1H), 7.03 (dd,
1 H), 4.56 (m, 1 H), 3.87(s, 3H), 3.84 (m, 2H), 3.76 (s, 2H),1.85 (m, I H), I
.30
(d, 3H) 0.86 (d, 6H).
C,gH,3NO,,S (MW = 349, Mass Spectroscopy (MH' 350)).
Example B55
Synthesis of N [(3-phenyl-1,2,4-thiadiazol-5-yl)acetyl] alanine iso-butyl
ester
Following General Procedure BI above, and using (3-phenyl-1,2,4-
thiadiazoI-5-yl)acetic acid (CAS# 90771-06-5 ) and alanine iso-butyl ester
(prepared following General Procedure BJ above), the title compound was
prepared. The reaction was monitored by tlc on silica gel and purification was
by filtration as described in the general procedure.
NMR data was as follows:
'H-nmr (CDCI,): 8 = 7.47 (m, 5H), 4.66 (m, 1H), 4.16 (s, 2H), 3.91 (m,
2H), 1.93 (m, 1H), 1.48 (d, 3H) 0.93 (d, 6H).
C"H2,N303S {MW = 347, Mass Spectroscopy (MH+ 348)).
Example B56
Synthesis of N [2-phenyloxazol-4-yl)acetyl]alanine iso-butyl ester
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Following General Procedure BI above, and using (2-phenyloxazol-4-
yl)acetic acid (CAS# 22086-89-1 ) and alanine iso-butyl ester (prepared
following
General Procedure BJ above), the title compound was prepared. The reaction
was monitored by tlc on silica gel and purification was by filtration as
described
in the general procedure.
Example B57
Synthesis of N-[(3-methylphenyl)acetyi]alanine iso-butyl ester
Following General Procedure BI above, and using 3-methylphenylacetic
acid (Aldrich) and alanine iso-butyl ester (prepared following General
Procedure
BJ above), the title compound was prepared. The reaction was monitored by tlc
on silica gel and purification was by filtration as described in the general
procedure.
NMR data was as follows:
' H-nmr (CDCI,): 8 = 7.21 (m, 1 H), 7.07 (m, 3H), 4.54 (m, 1 H), 3.83 (m,
2H), 3.52 (s, 2H), 2.35 (s, 3H), 1.87 (m, IH), 1.32 (d, 3H), 0.88 (d, 6H).
C,6H,3NO3 (MW = 277, Mass Spectroscopy (MH+ 278)).
Example B58
Synthesis of N [{2,5-difluorophenyl)acetyl]alanine iso-butyl ester
Following General Procedure BI above, and using 2,5-
difluorophenylacetic acid (Aldrich) and alanine iso-butyl ester (prepared
following General Procedure BJ above), the title compound was prepared. The
reaction was monitored by tlc on silica gel and purification was by filtration
as
described in the general procedure.
NMR data was as follows:
'H-nmr (CDCI,): 8 = 7.08-6.94 (m, 3H), 4.57 (m, 1H), 3.91 (m, 2H),
3.56 (s, 2H), 1.92 (m, 1H), 1.41 (d, 3H) 0.91 (d, 6H).
C'SH'9N03Fz (MW = 299, Mass Spectroscopy (MH+ 300)).
Example B59
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Synthesis of N [(3,5-ditlurophenyl)acetyl(aianine iso-butyl-ester
- Following General Procedure BI above, and using 3,5-
' difluorophenylacetic acid {Aldrich) and alanine iso-butyl ester (prepared
following General Procedure BJ above), the title compound was- prepared. The
S reaction was monitored by tlc on silica gel and purification was by
filtration as
described in the general procedure.
NMR data was as follows:
'H-nmr (CDCI,): 8 = 6.81 (m, 2H), 6.74 (m, 1H), 6.06 (m, 1H), 4.57 (m,
1 H)) 3.92 (m, 2H), 3.51 (s, 2H), 1.94 (m, 1 H), 1.36 (d, 3H) 0.87 (d, 6H).
C,5H,9N03F~ (MW = 299, Mass Spectroscopy (Ml'I' 300)).
Example B60
Synthesis of N ((3-thienyl)acetyl]alanine iso-butyl ester
Following General Procedure BI above, and using 3-thiopheneacetic acid
(Aldrich) and alanine iso-butyl ester {prepared following General Procedure BJ
above), the title compound was prepared. The reaction was monitored by tlc on
silica gel and purification was by filtration as described in the general
procedure.
NMR data was as follows:
'H-nmr (CDCI,); cS = 7.33 (m, 1H), 7.14 (m, 1H), 7.01 (m, 1H), 6.09 (m,
1 H), 4.58 {m, 1 H), 3.88 (m, 2H), 3.60 (s, 2H), 1.91 (m, 1 H), 1.37 (d, 3H)
0.92
(d, 6H).
Optical Rotation: (ocJ,3 -52 (c 1 MeOH) @ 589 nm.
C,3H,9N03S (MW = 269, Mass Spectroscopy (MH+ 269)).
Example B61
Synthesis of N [{4-methylphenyl)acetyl]-L-alanine iso-butyl ester
Following General Procedure BI above, and using 4-methylphenylacetic
acid (Aldrich) and L-alanine iso-butyl ester {prepared following General
Procedure BJ above), the title compound was prepared. The reaction was
- 30 monitored by tlc on silica gel and purification was by filtration as
described in
the general procedure.
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NMR data was as follows:
'H-nmr (CDC1,): 8 = 7.11 {s, 4H), 5.93 (m, 1H), 4.58 (m, 1H), 3.88 (m,
2H), 3.54 (s; 2H), 2.33 (s. 3H), 1.89 (m, 1H), 1.32 (d, 3H), 0.89 (d, 6H).
C,6H,3NO3 (MW = 277.35, Mass Spectroscopy (MH + 278)).
Example B62
Synthesis of N (phenylacetyl)-L-alanine S-1-(methoxycarbonyl)
iso-butyl ester
Following General Procedure BK and using (S)-(+}-2-hydroxy-2-
methylbutyric acid (Aidrich) in place of the amino acid, methyl (S)-(+)-2-
hydroxy-2-methylbutyrate was prepared.
Methyl (S)-(+)-2-hydroxy-2-methylbutyrate was then coupled with
carbobenzyloxy-L-alanine (Aldrich) using General Procedure BE to provide
carbobenzyloxy-L-alanine S-1-(methoxycarbonyl) iso-butyl ester.
Carbobenzyloxy-L-alanine S-1-(methoxycarbonyl) iso-butyl ester (1.0 g)
was then dissolved in 20 mL of methanol and 6N HCl {0.5 mL) and 10%
palladium on carbon (0.1 g) were added. This reaction mixture was
hydrogenated at 40 psi of hydrogen on a Parr apparatus for 5 hours at room
temperature and then filtered through a pad of Celite. The filtrate was
concentrated at reduced pressure to provide L-alanine S-1-(methoxycarbonyl)
iso-butyl ester hydrochloride (98% yield}.
L-Alanine S-1-(methoxycarbonyl) iso-butyl ester hydrochloride was then
coupled to phenylacetic acid using General Procedure BG to provide the title
compound.
NMR data was as follows:
'H-nmr (CDCI,): 8 = 7.35 - 7.20 (m, 5H), 6.22 (bd, 1H}, 4.83 (d, 1H),
4.65 (p, 1H), 3.68 (s, 3H), 3.55 {s, 2H), 2.21 (m, 1H), 1.40 (d, 3H), 0.97 (d,
3H), 0.93 (d, 3H).
'3C-nmr {CDC13): 8 = 173.25, 171.18, 170.22, 135.11, 129.94, 129.50,
127.88, 52.67, 48.49, 43.98, 30.53, 19.21, 18.75, 17.58.
Example B63
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Synthesis of N-[(3-nitrophenyl)acetyl]-L-alanine iso-butyl-ester
Following General Procedure BH above and using 3-nitrophenylacetic
acid {Aldricfi) and L-alanine iso-butyl ester hydrochloride (from Example BB
above), the title compound was prepared. The reaction was monitored by tlc on
silica gel and purification was by recrystallization from butyl chloride.
NMR data was as follows:
'H-nmr (CDCI,): 8 = 8.17 (m, 2H), 7.68 (d, I H), 7.52 (t, 1 H), 6.18 (m,
1 H), 4.48 (m, I H), 3.94 (m, 2H), 3.67 (s, 2H), 1.93 (m, 1 H), 1.42 (d, 3H),
0.9I
(d, 3H).
Optical Rotation: [a],3 -49 (c 5, MeOH).
Example B64
Synthesis of N [(3,5-difluorophenyl)acetyl]alanine ethyl ester
Following General Procedure BG and using 3,5-difluorophenylacetic acid
(Aldrich) and alanine ethyl ester (Aldrich), the title compound was prepared
as a
solid with a melting point of 93°-95°C. The reaction was
monitored by tlc on
silica gel (Rf = 0.8 in EtOAC) and purification was by chromatography on
silica
gel using EtOAc as the eluant followed by recrystallization from I -
chlorobutane.
NMR data was as follows:
'H-nmr (DMSO-db): 8 = 1.30 (d, 3H); 3.52 (s, 2H).
C,3H,5N03F, (MW = 271.26, Mass Spectroscopy {MH~ 271)}.
Example B65
Synthesis of N [(3-nitrophenyl)acetyl]methionine ethyl ester
Following General Procedure BG above and using 3-nitrophenylacetic
acid (Aldrich) and methionine ethyl ester hydrochloride (Aldrich), the title
compound was prepared. The reaction was monitored by tlc on silica gel and
purification was by recrystallization from butyl chloride.
NMR data was as follows:
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' H-nmr (CDCI,): 8 = 8.18 (s, 1 H), 8.15 (d, 1 H) 7.66 (d, 1 H); 7.48- (t,
1 H), 6.3 0 (m, 1 H), 4.67 (m, I H), 4.21 (t, 2H), 3.67 (s, 2H), 2.47 (t, 2H),
2. I 2
(m, 2 H), 2.08 (s, 3H), 1.27 (t, 3H).
Optical Rotation: [a)~; -30 (c S, MeOH). -
Example B66
Synthesis of N [(3-chiorophenyl)acetyl]alanine iso-butyl ester
Following General Procedure BG above and using 3-chlorophenylacetic
acid (Aldrich) and alanine iso-butyl ester (prepared following General
Procedure
BJ above), the title compound was prepared. The reaction was monitored by tlc
on silica gel.
NMR data was as follows:
'H-nmr (CDCI,): b = 7.29 (m, 3H), 7.18 (m, 1H), 6.0 (m, 1H), 4.56 (m,
1 H), 3.89 (m, 2H), 3.53 (s, 2H), 1.91 (m, 1 H), 1.39 (d, 3 H), 0.91 (d, 3H).
Optical Rotation: [a.],3 -45 (c 5, MeOH).
C, SH.,oN03Cl (MW = 297.78, Mass Spectroscopy (MH+ 297)).
Example B67
Synthesis of N [(3-chlorophenyl)acetyl]alanine
2-(N,1V dimethylamino)ethyl ester
Following General Procedure BC above, and using N-(3-chlorophenyl-
acetyl)alanine (from Example BD above) and 2-(N,N-dimethyl amino) ethanol
{Aldrich), the title compound can be prepared. The reaction was monitored by
tlc on silica gel and purification was by liquid chromatography using
0.1:2:0.79
NH40H:EtOH:CHC13 as the eluant.
NMR data was as follows:
'H-nmr (CDCI,): 7.37 (s, 1H), 7.33-7.2 (m, 3H), 4.675-4.6 (m, 1H), 4.5-
4.37 (m, 1 H), 4.25-4.13 (m, 1 H), 3.6 (d, J = 7 Hz, 2H), 2.86 (bs, 2H), 2.3
(s,
6H), 1.23 (d, J = 9 Hz, 3H).
C,SH~,N~03C1 (MW = 313.799, Mass Spectroscopy (M+ 313)).
Example B68
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Synthesis of 2-[(3,5-dichlorophenyl)acetamido)hexanoic acid methyl ester
Following General Procedure BF above, an using 3,5-
dichlorophenylacetic acid (from Example BC above) and L-norleucine methyl
ester hydrochloride (Bachem), the title compound was prepared-as a solid
having
- 5 a melting point of 77°-78°C. The reaction was monitored by
tlc on silica gel
(Rf = 0.70 in 40% EtOAC/hexanes) and purification was by flash
chromatography on silica gel using 40% EtOAc/hexanes as the eluant.
NMR data was as follows:
'H-nmr (CDCI,): 8 = 7.20 (s), 7.18 (s), 6.6 (m), 4.55 (m), 3.7 (s), 3.5 (s),
3.4 (s), 2.0 (s}, I.8 (m), 1.6 (m}, 1.2 {m), 0.8 (t).
''C-nmr (CDC13): 8 = 173.54, 169.67, 138.43, 135.72, 128.33, 128.07.
78.04, 77.62, 77.19. 53.04, 52.90, 43.14, 32.57, 27.87, 22.81, 14.41.
Example B69
Synthesis of N [(3,5-diclorophenyl)acetyl)-L-alanine iso-butyl ester
Following General Procedure BF above, and using 3,5-
dichlorophenylacetic acid (from Example BC above) and L-alanine is~o-butyl
ester hydrochloride (from Example BB above), the title compound was prepared
as a solid having a melting point of 115°-116°C. The reaction
was monitored
by tlc on silica gel (Rf = 0.40 in 3% methanol/dichloromethane) and
purification
was by flash chromatography on silica gel using 3% methanol/dichloromethane
as the eluant.
NMR data was as follows:
'H-nmr (CDCI,): s = 7.27 (d, J = 2 Hz, 1 H), 7.19 (s, 2H), 6.22 (d, J =
6 Hz, 1 H), 4.59 (quint., J = 7 Hz, 1 H), 3.9 (q, J = 4 Hz, 2H), 3.5 (s, 2H},
I .9
(m, 1 H), 1.4 (d, J = 7 Hz, 3H), 0.91 (d, J = 7 Hz, 6H).
'3C-nmr (CDC13): b = 173.45, 169.37, 138.31, 135.75, 128.39, 128.11,
78.04, 77.61, 77.19, 72.19, 54.03, 48.97, 43.12, 28.24. 19.52, 19.49, I 9.09.
C,SH,9N03C1~ (MW = 331.9, Mass Spectroscopy (MH~ 332)).
Example B70
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Synthesis of N (cyclohexylacetyl)-L-alanine iso-butyl ester
Following General Procedure BB above, and using cyclohexyiacetic acid
(Aldrich) arid L-alanine i.so-butyl ester hydrochloride (from Example BB
above),
the title compound was prepared as a solid having a melting point of
92°C-
93°C. The reaction was monitored by tlc on silica gel {Rf = 0.39 in 1:3
EtOAc:hexane) and purification was by extraction with Et~O followed by washes
with aqueous K~C03 and aqueous HCI.
NMR data was as follows:
'H-nmr (CDCI,): 8 = 0.93 (d, J = 6.7 Hz, 6H), 0.85-1.01 (m, 2H), 1.05-
1.35 (m, 3H), I.40 (d, J = 7.I Hz, 3H), 1.60-1.85 (m, 6H), 1.95 (m, IH), 2.06
(d, J = 7.0 Hz, 2H), 3.92 (m, 2H), 4.61 (m, 1 H), 6.08 (bd, 1 H).
''C-nmr (CDCl3): 8 = 18.7, 18.9, 26.0, 26.1, 27.6, 33.0, 35.3, 44.6, 47.9,
71.4, 171.8, 173.3.
C,SH~,N03 {MW = 269.39, Mass Spectroscopy (MH+ 270)).
Example B71
Synthesis of N (cyclopentylacetyl)-L-alanine iso-butyl ester
Following General Procedure BB above, and using cyclopentylacetic acid
(Aldrich) and L-alanine iso-butyl ester hydrochloride (from Example BB above),
the title compound was prepared as a solid having a melting point of
62°C-
64°C. The reaction was monitored by tlc on silica gel (Rf = 0.37 in 1:3
EtOAc:hexane) and purification was by extraction with Et,O followed by washes
with aqueous KzC03 and aqueous HCI.
NMR data was as follows:
'H-nmr (CDCI,): 8 = 0.87 (d, J = 6.8 Hz, 6H), 1.01-1.17 (m, 2H}, 1.34
(d, J = 7.2 Hz, 3 H), 1.40-1.62 (m, 4H), 1.70-1. 83 (m, 2H), 1. 89 (m, 1 H),
2.15
(m, 3H), 3.86 (m, 2H), 4.55 (m, 1H), 6.30 (d, J= 7.1 Hz, IH).
'''C-nmr (CDC13): 8 = 18.4, 18.78, 18.80, 24.8 (very high), 27.5, 32.27,
32.32, 36.9, 42.5, 47.7, 71.2, 172.2, 173.2.
Elemental Analysis-Calc (%): C, 65.85; H, 9.87; N, 5.49; Found (%): C,
66.01; H, 10.08; N, 5.49.
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C~4HZSN03 (MW = 255.36, Mass Spectroscopy (MH+ 256)). -
Example B72
Synthesis of N [(cyclohex-1-enyl)acetyl]-L-alanine iso=butyl ester
Following General Procedure BB above, and using cyclohex-1-enyl acetic'
acid (Alfa) and L-alanine iro-butyl ester hydrochloride (from Example BB
above), the title compound was prepared as a solid having a melting point of
49°C-51 °C. The reaction was monitored by tlc on silica gel (Rf
= 0.40 in 1:3
EtOAc:hexane) and purification was by extraction with Et,O followed by washes
IO with aqueous K,C03 and aqueous HCI.
NMR data was as follows:
'H-nmr (CDCI,): 8 = 0.91 (d, J = 4.5 Hz, 3H), 0.93 (d, J = 6.7 Hz, 3H),
1.40 (d, J = 7.2 Hz, 3H), 1.52-1.70 (m, 4H), 1.97 (m, 3H), 2.06 (bs, 2H), 2.89
(s, 2H), 3.92 (m, 2H), 4.59 (m, 1 H), 5.65 (s, 1 H), 6.33 (d, J = 6.6 Hz, 1
H).
I5 '3C-nmr (CDCl3): 8 = 18.7, 18.91, 18.93, 21.9, 22.7, 25.3, 27.6, 28.3,
46.1, 47.9, 71.4, 127.1, 132.5, 170.6, 173.1.
Elemental Analysis-Calc (%): C, 67.38; H, 9.42; N, 5.24; Found (%): C,
67.34; H, 9.54; N, 5.16.
C,SH~SN03 (MW = 267.37, Mass Spectroscopy (MH+ 268)).
Example B73
Synthesis of N [(3-chlorophenyl)acetyl]alanine 3-methylbut-2-enyl thioester
Following General Procedure BC above, and using N [(3-
chlorophenyl)acetyl] alanine and 3-methyl-2-butene thioester (TCI), the title
compound can be prepared. The reaction was monitored by tlc on silica gel and
purification was by liquid chromatography using 3:7 EtOAc:Hexane as the
_ eluant.
NMR data was as follows:
'H-nmr (DMSO-db): 8 = 5.2-5.075 (m, 1H), 4.37 (dq, J = 9 Hz, 1H),
3.56 (s), 3.43 (d, J = 12 Hz, 2H), 1.266 (d, J = 12 Hz, ' 6H) 1.3 (d, J = 9
Hz,
3H).
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C,6HzoNO~CIS (MW = 325.86, Mass Spectroscopy (M+ 325)).
- Example B74
Synthesis of N [(2-phenyl)-2-fluoroacetyl]alanine ethyl ester
Following General Procedure BF above, and using oc-fluorophenyl acetic
acid (Aldrich) and alanine ethyl ester (Aldrich), the title compound was
prepared. The reaction was monitored by tlc on silica gel (Rf = 0.75 in l:l
EtOAc:hexane) and purification was by chromatography on silica gel using I:2
ethyl acetate/hexanes as the eluent.
NMR data was as follows:
'H-nmr (DMSO-db): b = 1.14 (d, 31-1), 1.34 (d, 3H), 4.07 (m, 2H), 4.33
(m, 1 H), 5.84 (d, 1 H), 6.01 (d, I H), 7.40-7.55 (m, SH), 8.87 (m, 1 H).
C,3H,6N03F (MW = 253.27, Mass Spectroscopy (MH+ 253)).
Example B75
Synthesis of N (3,5-difluorophenylacetyl)-L-phenylglycine methyl ester
Following General Procedure BF above, and using 3,5-
difluorophenylacetic acid (Aldrich) and L-phenylglycine methyl ester
hydrochloride (Bachem), the title compound was prepared.
NMR data was as follows:
'H-nmr (CDCl3): 8 =7.4-7.3 (m, 5H), 6.9-6.7 (m, 3H), 6.55 (d 1H, 7.1
Hz), 5.56 (d I H 7 Hz), 3.72 (s 3H), 3.57 (s 2H)
''C-nmr {CDC13): 8 = 197.6, 177.6, 171.8, 169.3, 136.7, 129.6, 129.3,
127. 8, 113.0, 112.9, 112.7, 111.4, 103. 8, 103.5, 65.1, 57.2, 53.5, 45.1, 43
.3,
43.3
C,.,H,SN03F, {MW = 319.31, Mass Spectroscopy (MH +320)).
Example B76
Synthesis of N (3,S-difluorophenylacetyl)-L-phenylglycine iso-butyl ester
The 3,5-difluorophenylacetic acid {Aldrich) was EDC coupled to L-
phenylglycine methyl ester hydrochloride (Bachem) via General Procedure BF
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above. The resulting compound was placed in a large excess of the desired
alcohol. A catalytic amount of dry NaH was added, and the reaction was
followed by-tlc until the presence of starting material was no longer
detected.
The reaction was quenched with a few milliliters of 1N HCI, and after a few
' S minutes of stirring saturated aqueous NaHCO, was added. The volume of the
reaction mixture was reduced on a rotary evaporator unti 1 the excess alcohol
was
removed and then the remaining residue was taken up in ethyl acetate and
additional water was added. The organic phase was washed with saturated
aqueous NaCI and dried over MgSO,. The solution was stripped free of solvent
on a rotary evaporator, and the crude product residue was then further
purified
by chromatography.
NMR data was as follows:
'H-nmr (CDC13): S = 7.35-7.3 (m 5I-I), 6.8-6.7 (m 3H) 6.60 (d IH, 7
Hz), 5.55 (d 1 H 7.1 Hz), 3.9 (m 2H), 3.60 (s 2H), 1.85 (m I H 7 Hz), 0.8 (q
6H
7 Hz)
''C-nmr (CDC13): 8 = 171.3, 169.3, 165.4, 138.5, 137.0, 129.5, 129.2,
127.6, 113.1, 113.0, 112.8, I 12. 7, 103 .8, 103.5, 103.2, 75.5, 5 7.2, 43.4,
43 .3,
28.2, 19.3
C,oH~~NO,F~ (MW = 361.39, Mass Spectroscopy (MH +362)).
Example B77
Synthesis of N (cyclopentylacetyl}-L-phenylglycine methyl ester
Following General Procedure BD above, and using cyclopentylacetic acid
(Aldrich) with L-phenylglycine methyl ester hydrochloride (Bachem) the title
compound was prepared.
NMR data was as follows:
'H-nmr (CDC13): 8 = 7.35 (s, 5H), 6.44 (bd, 1H), 5.6 (d, 1H), 3.72 (s,
3H), 2.24 (bs, 3H), 1.9-1.4 (m, 6H), 1.2-1,05 (m, 2H)
'3C-nmr (CDCI3): 8 = 172.3, 171.7, 136.7, 129.0, 128.6, 127.3, 56.2,
52.7, 42.5, 36.9, 32.40, 32.38, 24.8
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Example B78
Synthesis of N (cyclopentylacetyl)-L-alanine methyl ester
Following General Procedure BD above, and using cyclopentylacetic acid
(Aldrich) with L-alanine methyl ester hydrochloride (Sigma) the title compound
was prepared.
NMR data was as follows:
'H-nmr (CDCl3): b = 6.38 (d, 1H), 4.50 (m, 1H), 3.65 (s, 3H), 2.13 (bs,
3H), 1.80-1.00 {m (includes d at 1.30, 3H), 11H)
'3C-nmr (CDC13): 8 = 173.7, 172.5, 52.1, 47.6, 42.3, 36.8, 32.15, 32.14,
18.0
C"H,9N03 (MW = 213.28, Mass Spectroscopy (MH+ 214)).
Example B79
Synthesis of N (cyclopropylacetyl)-L-phenylglycine methyl ester
Following General Procedure BD above, and using cyclopropylacetic acid
{Aldrich) with L-phenylglycine methyl ester hydrochloride (Bachem), the title
compound was prepared.
NMR data was as follows:
'H-nmr (CDC13): 8 = 7.35 (m, 5H) 6.97 (bd, J = 7.2 Hz, IH) 5.59 (d,
J = 7.8 Hz, 1 H), 3.71 (s, 3 H), 2.17 {m, 2H), 1.05-0.95 (m, 1 H), 0.62 (m,
2H),
0.02 (m, 2H)
'3C-nmr (CDC13): 8 = 171.9, 174.6, 136.6, 129.0, 128.5, 127.2, 56.1,
52.7, 41.0, 6.9, 4.37, 4.33
Example B80
Synthesis of N-(cyclopropylacetyl)-L-alanine methyl ester
Following General Procedure BD above, and using cyclopropylacetic acid
(Aldrich) with L-alanine methyl ester hydrochloride (Sigma), the title
compound
was prepared.
NMR data was as follows:
_......~._, ,".~ . . .._._. . ,. .
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'H-nmr (CDC13): b = 6.60 (d, 1 H), 4.55 (m, 1 H), 3.69 (s, 3H), 2:10 (m,
' 2H), 1.34 (d, 3H), 0.95 (m, 1 H), 0.58 (m, 2H) 0.15 (m, 2H)
~3C-rimr (CDC13): 8 = 173.7, 172.3, 52.3, 47.7, 41.0, I8.2, 6.7, 4.27, 4.22
Example B81
Synthesis of N [(3-nitrophenyt)acetyl]-L-methionine iso-butyl ester
Following General Procedure BH above, and using nitrophenylacetic acid
(Aldrich) and L-methionine {Aldrich), the title compound was prepared as a tan
oil. The reaction was monitored by tlc on silica gel.
NMR data was as follows:
'H-nmr (CDCI3): 8 = 8.16 (m,2H) 7.67 (d, l I-I) 7.32 (t, 1 H), 6.31 (bd,
1 H), 4.69 (m, 1 H), 3 .90 (d, 2H), 3.68 (s, 2H), 2.47 (t, 2H), 2.15 (m, 1 H),
2.02
(s, 3H), 1.90 (m, 2H), 0.91 (d, 6H).
C"H24N~05S (MW = 368.4, Mass Spectroscopy (MH' 368)).
The following General Procedures and Examples illustrate the synthesis
of various Iactams and related compounds which can be used to prepare, for
example, compounds of formula VII and VIII above.
GENERAL PROCEDURE III-A
First EDC Coupling Procedure
To a l:l mixture of the corresponding carboxylic acid and the
corresponding amino acid ester or amide in CH,Ch at O°C was added 1.5
equivalents triethylamine, followed by 2.0 equivalents hydroxybenzotriazole
monohydrate and then 1.25 equivalents of ethyl-3-(3-dimethylamino)propyl
carbodiimide'HCI. The reaction mixture was stirred overnight at room
temperature and then transferred to a separatory funnel. The mixture was
washed with water, saturated aqueous NaHC03, 1 N HCl and saturated aqueous
NaCI, and then dried over MgSOa. The resulting solution was stripped free of
. 30 solvent on a rotary evaporator to yield the crude product.
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GENERAL PROCEDURE III-B -
Second EDC Cou~lin Procedure
A mixture of the corresponding acid (1 eqv), N-1-hydroxybenzotriazole
( 1.6 eqv), the corresponding amine ( 1 eqv), N-methylmorpholine ( 3 eqv) and
dichloromethane (or DMF for insoluble substrates) was cooled in an ice-water
bath and stirred until a clear solution was obtained. EDC ( 1.3 eqv) was then
added to the reaction mixture. The cooling bath was then allowed to warm to
ambient temperature over 1-2 h and the reaction mixture was stirred overnight.
The reaction mixture was then evaporated to dryness under vacuum. To the
residue was added 20% aqueous potassium carbonate and the mixture was
shaken throughly and then allowed to stand until the oily product solidified
(overnight if necessary). The solid product was then collected by filteration,
washed thoroughly with 20% aqueous potassium carbonate, water, 10% HC1,
and water to give the product, usually in pure state. No racemization was
observed.
GENERAL PROCEDURE III-C
Third EDC Counlin~Procedure
The carboxylic acid was dissolved in methylene chloride. The
corresponding amino acid ester or amide (1 eq.), N-methylmorpholine (5 eq.)
and hydroxybenzotriazole monohydrate ( 1.2 eq.) were added in sequence. A
cooling bath was applied to the round bottomed flask until the solution
reached
0°C. At that time, 1.2 eq. of 1-(3-dimethylaminopropyl)-3-
ethylcarbodiimide
hydrochloride was added. The solution was allowed to stir overnight and come
to room temperature under nitrogen pressure. The reaction mixture was worked
up by washing the organic phase with saturated aqueous sodium carbonate, 0.1 M
citric acid, and brine before drying with sodium sulfate. The solvents were
then
removed to yield crude product.
GENERAL PROCEDURE III-D
Fourth EDC Cowling Procedure
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A round bottom flask was charged with the corresponding carboxylic
° acid ( I .0 eq.), hydroxybenzotriazole hydrate ( 1.1 eq.) and the
corresponding
amine ( 1.0 eq. ) in THF under nitrogen atmosphere. An appropriate amount ( 1.
I
eq for free amines and 2.2 eq. for hydrochloride amine salts) of base, such as
Hunig's base was added to the well stirred mixture followed by EDC ( 1. I
eq.).
After stirring from 4 to 17 hours at room temperature the solvent was removed
at reduced pressure, the residue taken up in ethyl acetate (or similar
solvent) and
water, washed with saturated aqueous sodium bicarbonate solution, 1 N HCI,
brine, dried over anhydrous sodium sulfate and the solvent removed at reduced
pressure to provide the product.
GENERAL PROC'.EDURE III-E
BOP Couplin,Q Procedure
To a stirred solution of N (3,5-difluorophenylacetyl)alanine (2 mmol} in
DMF, cooled in an ice-water bath, was added BOP (2.4 mmol) and N-
methylmorpholine (6 mmol). The reaction mixture was stirred for 50 min. and
then a solution of a-amino-y-lactam (2 mmol) in DMF cooled at 0 °C was
added. The cooling bath was allowed to warm to ambient temperature over 1-2
h and the reaction mixture was then stirred overnight. A 20% aqueous
potassium carbonate solution {60 mL) was added and this mixture shaken
throughly. No solid formed. The mixture was then washed with ethyl acetate
( 150 rnL) and evaporated to dryness under vacuum to give a white solid. Water
(50 mL) was then added and this mixture shaken throughly. The precipitate that
formed was collected by filtration, then washed thoroughly with water,
followed
by 1 mL of diethyl ether to give the product (51 mg, 0.16 mmol, 7.8%).
GENERAL PROCEDURE III-F
Coupling of an Acid Chloride with an Amino Acid Ester
To a stirred solution of (D,L)-alanine isobutyl ester hydrochloride (4.6
mmol) in 5 ml of pyridine was added 4.6 mmol of the acid chloride.
Precipitation occurred immediately. The mixture was stirred for 3.5 h,
dissolved
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in 100 mL of diethyl ether, washed with 10% HCl three times, brine once, 20%
potassium carbonate once and brine once. The solution was dried over
magnesium sulfate, filtered, and evaporated to yield the product. Other amino
acid esters may also be employed in this procedure. -
GENERAL PROCEDURE III-G
Coupling of a Carboxylic Acid with an Amino Acid Ester
A solution of the carboxylic acid (3.3 mmol) and 1,1'-carbodiimidazole
(CDI) in 20 mL THF was stirred for 2 h. (D,L)-alanine isobutyl ester
hydrochloride (3.6 mmol) was added, followed by 1.5 mL (10.8 mmol) of
triethylamine. The reaction mixture was stirred overnight. The reaction
mixture
was dissolved in 100 mL of diethyl ether, washed with 10% HC1 three times,
brine once, 20% potassium carbonate once and brine once. The solution was
dried over magnesium sulfate, filtered, and evaporated to yield the product.
Other amino acid esters may also be employed in this procedure.
GENERAL PROCEDURE III-H
Fifth EDC Coupling Procedure
In a round bottom flask was added a carboxylic acid ( I .1 eq.) in THF, an
amine hydrochloride ( I .0 eq. ), 1-hydroxybenzotriazole hydrate ( 1.1 eq. ),
N,N-
diisopropylethylamine {2.1 eq.), followed by 1-{3-dimethylaminopropyl)-3-
ethylcarbodiimide hydrochloride (EDC) ( I .1 eq.). The reaction mixture
stirred
at room temperature for 10-20 hours under an atmosphere of nitrogen. The
mixture was diluted with EtOAc and washed with 0.1 M HCl ( 1 x 10 mL),
saturated NaHC03 { 1 x 10 mL), HBO ( 1 x 10 mL), and brine and dried over
MgSO~. The drying agent was removed by filtration and the filtrate was
concentrated in vacuo. The residue was purified by flash column
chromatography on silica gel followed by trituration from EtOAc and hexanes.
GENERAL PROCEDURE III-I
Sixth EDC Couplin,~ Procedure
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To a solution or suspension of the amine or amine hydrochloride (1.0
eq.) in THF (0.05-0. I M) under N, at 0°C was added the carboxylic acid
( 1.0-
1.1 eq.), hydroxybenzotriazole monohydrate (I.I-1.15 eq.), Hunig's base (I.I
eq.
for free amines and 1.1-2.3 eq. for hydrochloride amine salts), followed by I -
(3-
dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride ( I . I -1. I 5 eq.).
The
cooling bath was removed and the mixture allowed to warm to room
temperature for 10-24 hours. The solution or mixture was diluted with EtOAc,
in a 3-5 volume multiple of the initial THF volume, and washed with 0.1-1.0 M
aq. HCl ( 1 or 2x), dilute NaHC03 ( 1 or 2x), and brine ( I x). Then, the
organic
phase was dried over either MgS04 or Na,SO~, filtered, concentrated to provide
the crude product, which was either further purified or utilized without
further
purification.
GENERAL PROCEDURE III-J
EEDO Couple Procedure
To a solution of the amine in THF (1.0 eq., 0.05-0.08 M, final molarity)
under N, at room temperature was added the N-t-Boc protected amino acid ( I .1
eq., either as a solid or in THF via cannula), followed by EEDQ (Aldrich, 1.1
eq.). The pale yellow solution was stirred at room temperature for 16-16.5
hours, then diluted with EtOAc (in a 3-5 volume multiple of the initial THF
volume), and washed with 1 M aq. HCI (2x), dilute aq. NaHC03 (2x), and brine
( I x). The organic phase was dried over either Na~S04 or MgS04, filtered, and
concentrated.
. 25 Example 2-A
Synthesis of
5-Amino-5,7-dihydro-6H-
dibenzo[a,c]cyclohepten-6-of Hydrochloride
Step A - Synthesis of 5-Oximo-5,7-dil~dro-6H-
dibenzoja,clcyclohepten-6-one
A round bottom flask was charged with 5,7-dihydro-6H-
dibenzo[a,c]cyclohepten-6-one ( I .0 g, 4.81 mmol)(CAS# I 139-82-8, prepared
as
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described in Tetrahedron Letters, Vol. 28, No. 23, (1987), pp 2633-2636) and
butyl nitrite (0.673 ml, 5.77 mmol) (Aldrich) in Et-,O. The solution was
cooled
to 0°C and treated drop-wise with a saturated solution of HCl(g)/Et20.
After 5
h at 0°C the resulting precipitate was filtered, rinsed with cold Et,O
and vacuum
dried to give the title compound as a colorless solid.
NMR data was as follows:
'H-nmr (CDCl3): d = 7.26-7.74 (m, 8H), 3.84 (m, 2H).
C,SH"NO, {MW = 237.26); mass spectroscopy (MH+) 238.
Anal. Calcd for C,SH"NO,; C, 75.93 H, 4.67 N, 5.90. Found: C, 75.67
H, 4.83 N, 5.67.
Step B- Synthesis of 5-Amino-5,7-dihydro-6H-
dibenzofa,c,Lcyclohepten-6-of Hydrochloride
The compound isolated above (0.489 g, 2.04 mmol) was dissolved in
THF and added drop-wise to a well-stirred mixture of LAH ( 10.2 ml, 10.2
mmol)/THF. After heating to reflux for 25 h under N, atmosphere the solution
was quenched and worked-up according to Fieser's method. The resulting solid
was rinsed with NH3 sat/CHC13, the filtrate evaporated and the title compound
purified by chromatography (SiO,. CHC13}.
C,SH,SNO (MW = 225.290); mass spectroscopy (MH+) 226.
Anal. Calcd for C,SH,SNO; C, 79.97 H, 6.71 N, 6.22. Found: C, 80.19
H, 6.71 N, 5.91.
GENERAL PROCEDURE S-A
N-Alkylation of Lactams
To a stirred solution of a BOC-protected a-aminocaprolactam (6.87 g, 30
mmol) in DMF (150 mL) was added in portions 97% NaH (1.08g, 45 rnmol).
Bubbling occured immediately and followed by heavy precipitation. After 10
min., benzyl bromide (3.93 mL, 33 mmol) was added. The precipitate dissolved
quickly and in about 10 min. a clear solution was obtained. The reaction
mixture was stirred overnight and then evaporated as completely as possible on
a
rotovap at 30°C. Ethyl acetate (100 mL) was added to the residue and
this
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mixture was washed with water, brine, and dried over magnesium sulfate. After
filtration and concentration, a thick liquid ( 10 g) was obtained which was
then
chromatographed over silica gel with 1:3 ethyl acetate/hexane as the eluant to
provide 5.51 g (58%) of the N-benzylated product as an oil. Other lactams and
- 5 alkylating agents may be used in this procedure to obtain a wide variety
of N-
alkylated lactams. Various bases, such as LiN(SiMe3), may also be employed.
GENERAL PROCEDURE 5-B
BOC Removal Procedure
The BOC-protected compound in a 1:1-2:1 mixture of CH~CI, and
trifluoroacetic acid was stirred until tlc indicated complete conversion,
typically
2 hours. The solution was then stripped to dryness and the residue was taken
up
in ethyl acetate or CH,Cl2. The solution was washed with saturated aqueous
NaHC03 and the aqueous phase was adjusted to a basic pH, then extracted with
ethyl acetate or CH~C1~. The organic phase was washed with saturated aqueous
NaCI and dried over MgS04. The solution was stripped free of solvent on a
rotary evaporator to yield the product.
GENERAL PROCEDURE S-C
Synthesis of a-Aminolactams
The Schmidt reaction was conducted on 4-ethylcyclohexanone using
hydroxyamine sulfonic acid as described in Olah, Org. Synth. Collective, Vol.
VII, page 254, to provide 5-ethylcaprolactam in 76% yield. Using the procedure
described in Watthey, et al., J. Med. Chem. , 1985, 28, 1511-1516, this lactam
was then dichlorinated with PC15 at the alpha position and reduced by
hydrogenation to provide four isomeric monochlorides (two racemic mixtures}.
The two racemic mixtures were separated from each other by column
chromatography using silica gel and each racemic mixture was reacted with
sodium azide to yield the corresponding azide which was hydrogenated to
provide the corresponding a-aminolactams. Other cycloalkanones may be
employed in this procedure to provide a wide variety of a-aminolactams. In
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some cases, such as when preparing the 9-membered ring a-aminolactam~ longer
reaction times, higher reaction temperatures and an excess of sodium azide may
be required. ' For example, the 9-membered ring a-aminolactam required 5
equivalents of sodium azide, a reaction temperature of 120°C arid a
reaction
time of 4 days. Such conditions can be readily determined by those of ordinary
skill in the art.
GENERAL PROCEDURE 5-D
Synthesis of 4-Amino-1,2,3,4-tetrahydroisoquinoline-3-ones
The 4-amino-1,2,3,4-tetrahydroisoquinoline-3-one derivatives employed
in this invention can be prepared by the following art-recognized procedures.
The conditions for these reactions are further described in D. Ben-Ishai, et
al.,
Tetrahedron, 43, 439-450 ( 1987). The following intermediates were prepared
via this procedure:
3-amino-1,2, 3,4-tetrahydroisoquinolin-3-one
4-amino-7-benzyl-1,2,3,4-tetrahydroisoquinolin-3-one
4-amino-1-phenyl-1,2,3,4-tetrahydroisoquinolin-3-one
cis and traps-4-amino-I -phenyl- I ,2,3,4-tetrahydroisoquinolin-3-one
4-amino-2-phenethyl-1,2,3 ,4-tetrahydroisoquinolin-3-one
4-amino-2-methyl-1,2,3,4-tetrahydroisoquinolin-3-one
9-amino(fluoren-I-yl)glycine 8-lactam-1,2,3,4-tetrahydroisoquinolin-3-
one.
Step A - Preparation of N-Bismethoxycarbonylaminoacetic Acid: To one
mole equivalent of glyoxylic acid in 2 liters of ethanol-free chloroform was
added two mole equivalents of methyl carbamate and 0.1 mole equivalent of
naphthalene sulfonic acid. The reaction mixture was then brought to a reflux
for
6 hours. Water was removed using an inverse Dean Stark trap. The reaction
was then cooled and the product filtered and washed with chloroform. The
white solid was recrystallized from ethyl acetate/hexanes to give a white
powder
in 65% yield.
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Step B - Coupling Procedure: To 0.0291 moles of N- -
bismethoxycarbonylaminoacetic acid (or the appropriate carboxcyclic acid) in
200 mL of T'HF was added one mole equivalent of EDC~HCI, a benzylamine,
HOBT, and diisopropylethylamine. The reaction was allowed to stir at room
temperature for 18 hours and then poured into a separatory funnel and
extracted
into ethyl acetate. The ethyl acetate solution was washed with 1 molar K~C03
and then 1 molar HC1. The organic layer was dried over Na~S04, filtered and
solvent removed to give the crystalline benzylamide of N-
bismethoxycarbonylaminoacetic acid. This material was used without further
purification. Typical yields range from 40 - 55%.
Step C - Cyclization Procedure: The benzyiamide of N
bismethoxycarbonylaminoacetic acid (0.008 moles) was dissolved in 75 mL of
methanesulfonic acid and allowed to stir over night at room temperature. The
reaction mixture was poured over ice and extracted into ethyl acetate. The
ethyl
acetate extract was washed with 1 molar K,CO~ and then 1 N HCI. The organic
layer was dried over Na~S04, filtered and the solvent removed to give the
crystalline 4-methoxycarbonylamino-1,2,3,4-tetrahydroisoquinoline-3-one in 50-
90% yield. This material was used without further purification.
Sten D - Removal of the Metho~oxycarbonyl Group~,MOCO To the 4-
methoxycarbonylamino-1,2,3,4-tetrahydroisoquinoline-3-one (3.4 mmoles) in 30
mL of acetonitrile was added 2 mole equivalents of trimethylsilyliodide
(TMSI).
The reaction mixture was heated to 50-80°C for 3 hrs and then
cooled and
poured into a seperatory funnel. The reaction mixture was diluted with ethyl
acetate and washed with 1 molar KZC03 and then with 5% NaHS03. The
- organic layer was dried over Na,S04 and filtered. The solvent was removed
under reduced pressure to give the 4-amino-1,2,3,4-tetrahydroisoquinoline-3-
one
derivative. Typical yields range from 50-87%.
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Step E - Alternative Procedure for Removal of the Methoxyoxycarbonyl
Group: To 3.8 mmoles of the MOC-protected compound was added 10 mL of
30% HBr iri acetic acid and this reaction mixture was heated to 60°C
for 3 hrs.
The mixture was then cooled and hexanes were added. The hexanes layer was
decanted off and the residue as placed under reduced pressure to give a tan
solid. This solid was slurried in ether and filtered to give the 4-amino-
1,2,3,4-
tetrahydroisoquinoline-3-one hydrobromide salt. Typical yields range from 57-
88%.
Example 5-A
Synthesis of
3-Amino-1,2,3,4-tetrahydroquinolin-2-one
Step A: Sodium (0.30g, 1 IOM%) was added to anhydrous ethanol (45
mL) and the reaction mixture was stirred until homogenous. Diethyl N-
i5 acetylaminomalonate (2.SI g, 100 M%) was added in one portion and this
mixture was stirred for 1 h. 2-Nitrobenzyl bromide {2. Sg, l OOM%) was then
added in one portion and the reaction mixture was stirred for 3 h. The
reaction
was poured into water and extracted with ethyl acetate (3x) and then
backwashed with water (3x) and brine (lx). Treatment with MgS04,
rotoevaporation, and chromotography (30% EtOAc/hexanes) yielded diethyl N-
acetylamino-2-nitrobenzylmalonate in 82% yield.
Step B: Diethyl N-acetylamino-2-nitrobenzylmalonate ( 1 g, 1 OOM%) was
dissolved in a minimum amount of EtOH. Pd/C ( 10%, O.OSg) was added and
the reaction mixture was subjected to 50 psi of H~ for 3 hours. The reaction
was then filtered thru a pad of celite. Additional EtOH (25mL) and TsOH
(catalytic amount, 0.01 g) were added and this mixture was refluxed for 2
hours.
The reaction was rotoevaporated to a residue and then partitioned between
water
and ethyl acetate. The water layer was extracted with ethyl acetate {3x) and
the
combined ethyl acetate extracts were washed with water (3x) and then brine
(lx). Treatment with MgS04 and rotoevaporation yielded pure 3-(N-
acetylamino)-3-carboethoxy-1,2,3,4-tetrahydroquinolin-2-one (89% yield).
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Step C: 3-(N-Acetylamino)-3-carboethoxy-1,2,3,4-tetrahydroquinolin-2-
' one (0.75 g, 100M%) was suspended in 6N HCl (25 mL) and the mixture was
heated to 100°C for 3 hours. The reaction was cooled, rotoevaporated to
a
residue and then partitioned between water and ethyl acetate. The water was
' S extracted with ethyl acetate (3x) and the combined ethyl acetate extracts
were
then washed with water (3x) and then brine ( 1 x). Treatment with MgS04
followed by rotoevaporation yielded 3-(R,S)-amino- I ,2,3,4-tetrahydroquinolin-
2-
one (72% yield).
Example 5-B
Synthesis of
4-Amino-1-(pyrid-4-yl)-1,2,3,4-tetrahydroisoquinolin-3-one
Step A: To a solution of 4-cyanopyridine (Aldrich) (0.150 moles) in 300
mL of dry ether was added 1.1 eq, of phenylmagnesium bromide (Aldrich)
dropwise. The reaction was refluxed for 2 hours and then stirred overnight at
room temperature. Sodium borohydride ( 1.0 eq.) was added dropwise as a
solution in 200 mL of methanol (CAUTION -- very exothermic). The reaction
was then heated to reflux for 6 hours, cooled and quenched with a saturated
solution of ammonium chloride. The solution was decanted from the salt in the
reaction mixture and acidified with 1N HCI. After washing the aqueous layer
with ethyl acetate, the pH of aqueous layer was adjusted to about 9.0 with 1 N
sodium hydroxide (cold). The aqueous layer was then extracted with ethyl
acetate and the organic extracts washed with brine, dried over Na,S04,
filtered
and concentrated to give 4-pyridyl-a,-benzyl amine as a thick yellow oil.
Steo B: Following General Procedure 5-D and using 4-pyridyl-a,-benzyl
amine, the title compound was prepared.
Example 5-C
Synthesis of
. 4-Amino-1-(pyrid-2-yl)-1,2,3,4-tetrahydroisoquinolin-3-one
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Step A: 2-Pyridyl-a-benzyl amine was' prepared by substituting 2-
cyanopyridine (Aldrich) for 4-cyanopyridine in the procedure described in
Example 5-B.
Step B: Following General Procedure 5-D and using 4-pyridyl-a-benzyl
amine, the title compound was prepared.
Example 5-D
Synthesis of
4-Amino-1-(pyrid-3-yl)-1,2,3,4-tetrahydroisoquinolin-3-one
St-ep A: Following the procedure described in J. Med. Chem., 1982, 25,
1248, and using 3-benzoyl-pyridine (Aldrich), 3-pyridyl-oc-benzyl amine was
prepared.
Step B: Following General Procedure 5-D and using 3-pyridyl-a.-benzyl
amine, the title compound was prepared.
Example 5-E
Synthesis of
4-Amino-7-benzyl-1,2,3,4-tetrahydroisoquinolin-3-one
Step A: To a Parr bottle containing 3-benzoylbenzoic acid (0.044 moles)
(Aldrich) in 150 mL of ethyl acetate and 4.5 mL of concentrated H,SO4 was
added 10 grams of 5% Pd/C. The mixture was hydrogenated on a Parr
apparatus under hydrogen (45 psi) overnight. The reaction mixture was then
filtered through Hyflo, washing with ethyl acetate. The filterate was dried
over
Na,S04, filtered and concentrated to give an oil. The oil was slurried in
hexane
and the resulting white solid was collected by filtration to afford 3-
benzylbenzoic acid, which was used without further purification.
Step B: To the product from Step A (0.0119 moles) was added 150 mL
of CH~Cl2, one drop of DMF, 10 mL of oxalyl chloride. and the mixture was
stirred at room temperature for 3 hours. After cooling to 10°C, 30 mL
of
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NH40H (exothermic) was added and the mixture was stirred for 30 min. The
' reaction mixture was then concentrated and the resulting residue diluted
with
ethyl acetate: The organic layer was washed with 1N NaOI-I, brine, dried over
Na,S04, and concentrated to give the 3-(benzyl)benzamide as a white solid,
' S which was used without further purification.
Step C: To a solution of 3-(benzyl)benzamide (.0094 moles) from Step
B in 70 of toluene was added 8 mL of Red-Al~ (65+ wt. % solution of sodium
bis(2-methoxyethoxy)aluminum hydride in toluene, Aldrich) (CAUTION --
reaction very exothermic). The reaction mixture was then heated at 60°C
for 2
hours and then poured over ice. The resulting mixture was extracted with ethyl
acetate and the combined extracts were washed with water and brine. The
organic layer was extracted with 1 N HCl and the aqueous layer washed with
ethyl acetate. The pH of the aqueous layer was then adjusted to about 9.0 with
1 N NaOH and extracted with ethyl acetate. The organic extracts were washed
with water and brine and then concentrated to give 3-(benzyl)benzyl amine.
Step D: Following General Procedure 5-D and using 3-(benzyl)benzyl
amine, the title compound was prepared.
Example 5-F
Synthesis of
4-Amino-6-phenyl-1,2,3,4-tetrahydroisoquinolin-3-one
Step A: To a solution of 4-biphenylcarboxamide (Aldrich) (0.025 mole)
in 150 mL of THF cooled to 10°C was added a solution of 1.5 eq of LAH (
1 M
in THF) dropwise. The reaction mixture turned from a white slurry to a green
homogenous solution and then to a yellow homogeneous solution. The reaction
was then quenched with 2.5 mL of 1 N NaOH. The mixture was then filtered
through Hyflo and extracted with ethyl acetate. The organic layer was then
washed with 1N HCI. The pH of the resulting aqueous layer was adjusted to
about 9 with 1 N NaOH and extracted with ethyl acetate. The organic extracts
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were washed with water and brine, and then dried over Na,SO~, filtered and
concentrated to give 4-(phenyl)benzyl amine as a white solid.
St_ ep B: Following General Procedure 5-D and using 4-(phenyl)benzyl
amine, the title compound was prepared.
Example 5-G
Synthesis of
cis- and traps-4-Amino-1-phenyl-1,2,3,4-tetrahydroisoquinolin-3-one
Step A: Following General Procedure 5-D and using a.-
phenylbenzylamine {Aldrich), 4-amino-1-phenyl-1,2,3,4-tetrahydroisoquinolin-3-
one was prepared.
Step B: To a solution of 4-amino-1-phenyl-1,2,3,4-tetrahydroisoquinolin-
3-one (0.00158 moles) from Step A in 20 mL of CH,C1, was added 2.0 eq. of
triethylamine and Boc anhydride (l.l eq.). The reaction was stirred overnight
at
room temperature and then concentrated. The residue was diluted with ethyl
acetate and water. The pH of the aqueous layer was adjusted to 3.0 with
sodium bisulfate and the layers were separated. 'The organic layer was dried
over Na~S04, filtered and concentrated. The residue was purified by LC 2000,
eluting with ethyl acetate/hexanes (70:30) to give a white solid containing a
1:1
mixture of cis- and traps-4-(N-Boc-amino)-1-phenyl-1,2,3,4-
tetrahydroisoquinolin-3-one isomers. This mixture was recrystallized from
ethyl
acetate to give the pure traps isomer and a cis isomer-enriched mixture of cis
and traps isomers. This mixture was recrystallized again from ethyl
acetate/hexanees (70:30) to give the pure cis isomer.
Step C: The cis isomer and the traps isomer from Step B were
separately deprotected using General Procedure 8-J to give cis-4-amino-1-
phenyl-1,2,3,4-tetrahydroisoquinolin-3-one and traps-4-amino-1-phenyl-1,2,3,4-
tetrahydroisoquinolin-3-one.
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Example S-H -
Synthesis of
. -4-Amino-7-phenyl-1,2,3,4-tetrahydroisoquinolin-3-one
Step A: To a solution of 1-bromo-3-phenylbenzene (Aldrich) (0.0858
moles) in 300 mL of dry THF cooled to -78°C was added tert-butyl
lithium (2
eq.) ( 1.7M in hexane) dropwise. The reaction mixture was stirred for 40 min.
at
-78°C and then quenched with 2 eq. of DMF ( 13.24 mL). The resulting
mixture
was stirred for 20 min. and then poured into a separatory funnel and extracted
with CH,Ch. The organic extracts were washed with water, dried over Na,S04,
filtered and concentrated to give a brown oil. This oil was purified by LC
2000
chromatography, eluting with ethyl acetate!hexanes (5:95) to give 3-
biphenylcarboxaldehyde.
Step B: To a solution of 3-biphenylcarboxaldehyde (0.011 eq.) in 30 mL
of methanol was added 10 eq. of 7N NH3/MeOH and NaCNBH4 (2 eq.). A
yellow gum precipitated from solution. The solution was then heated at
60°C
until gum dissolved and the solution was stirred at room temperature
overnight.
The reaction mixture was then concentrated and the resulting residue diluted
with ice water and ethyl acetate. The organic layer was then washed with brine
and extracted with SN HCI. The pH of the aqueous layer was then adjusted to
12 and the aqueous layer was extracted with cold ethyl acetate. The organic
layer was dried over Na,S04, filtered and concentrated to give 3-
(phenyl)benzyl
amine as an oil.
Step C: Following General Procedure 5-D and using 3-(phenyl)benzyl
amine, the title compound was prepared.
Example 5-I
Synthesis of
4-Amino-1-benzyl-I,2,3,4-tetrahydroisoquinolin-3-one
Step A: To a solution of benzoyl chloride (0.123 moles) (Aldrich) in
600 mL of CH,Ch was added 2.0 eq. of phenethylamine (Aldrich) dropwise.
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The reaction mixture was stirred at room temperature for 3 hours and then
poured into a separatory and extracted with CH,CI~. The organic extracts were
washed with-water and 1N HCI, and then dried over Na,S04, filtered and
concentrated to give N-phenethyl benzamide.
Step B: Reduction of N-phenethyl benzamide using the procedure of
Example 5-E, Step C afforded N-benzyl-N-phenethylamine as an oil.
Ste~C: Following General Procedure 5-D and using N-benzyl-N-
phenethylamine, the title compound was prepared.
Example ~-.1
Synthesis of
3-Amino-1-methyl-2-indolinone Monohydrochloride
St, ep A: {2,3-Dihydro-1-methyl-2-oxo-1 H-indol-3-yl)carbamic acid
methyl ester (CAS No. 110599-56-9) was prepared using the procedure
described in Ben-Ishai, D.; Sataty, I.; Peled, I~.; Goldshare, R. Tetrahedron
1987, 43, 439-450. The starting materials for this preparation were N-
methylaniline (CAS# 100-61-8, Eastman Kodak Co.), glyoxylie acid (CAS# 298-
12-4, Aldrich), and methyl carbamate (CAS# 598-55-0, Aldrich).
Step B: The product from Step A (333.5 mg) in 31% HBr in AcOH (10
mL) was heated to 50-60°C for 2 hours. The resulting orange solution
was
concentrated to a thick orange oil which was dissolved in EtOAc ( 15 mL) and
the product extracted into 1 M aq. HCl (10 mL). The aqueous acid was
neutralized with aq. NaHC03 and the product extracted into CHZCIz ( 10 x I 0
mL). _HCl (gas) was passed through the combined CH,CIZ extracts to form a
purple solution. The solution was concentrated to provide the title compound
(262.8 mg) as a purple solid.
Example 5-K
Synthesis of
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3-Amino-1-methyl-4-phenyl-3,4-traps-dihydrocarbostyril/Tin Complex
Step A: - Synthesis of 4-Phenyl-3,4-dihydrocarbostyril
4-Phenyl-3,4-dihydrocarbostyril (CAS# 4888-33-9) was prepared in two
steps using the procedure described by Conley, R. T.; Knopka, W. N. J. Org.
' S Chem. 1964, 29, 496-497. The starting materials for this preparation were
cinnamoyl chloride (Aldrich) and aniline (Aldrich). The title compound was
purified by flash chromatography eluting with CH,C1,/EtOAc (4: I ).
Step B: - Synthesis of 1-Methyl-4-phenyl-3,4-dihydrocarbostyril
To a suspension of NaH {I.2 eq., 0.537 g of 60% dispersion in mineral
oil) in THF (50 mL) under N~ at 0°C was added the product from Step A
(1.0
eq., 2.50 g) in THF (50 mL) via cannula over a period of 5 minutes. The
resulting pale yellow mixture was stirred at 0°C for 10 minutes, then
MeI (2.0
eq., 1.39 mL) was added. The opaque yellow mixture was allowed to slowly
{ice bath not removed) warm to ambient temperature with stirring for 15 hours.
1 M Aq. HC1 (50 mL) and EtOAc (250 ml,) were added and the phases
partitioned. The organic phase was washed with dilute NaHC03 ( 1 x 100 mL),
brine ( 1 x 100 mL), then dried over MgSO~, filtered, concentrated, and the
residue purified by flash chromatography eluting with CH,CI,/EtOAc ( 19:1
gradient to 15:1 ) to provide 1-methyl-4-phenyl-3,4-dihydrocarbostyril.
Step C: - Synthesis of 3-Azido-1-methyl-4-phenyl-3,4-trans-
dihydrocarbostyril
Following General Procedure 8-K, 3-azido-1-methyl-4-phenyl-3,4-trans-
dihydrocarbostyril was prepared as a white solid. The product was purified by
flash chromatography eluting with CH,C1,/hexanes/EtOAc 15:15:1.
Selected ~H-NMR data for the title compound (CDCl3): 8 = 4.46 (d, 1 H,
J = 10.57 Hz), 4.18 (d, IH, J = 10.63 Hz).
Step D: - Synthesis of 3-Amino-1-methyl-4-phenyl-3,4-trans-
dihydrocarbostyril/Tin Complex
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To a mixture of SnCI, (350.7 mg) in M-eOH (7 mL) under Nz at 0°C
was
added the product from Step C (257.4 mg) in MeOH/THF (5 mL/5 mL) via
cannula over a period of 1 minute. The cooling bath was removed the solution
allowed to warm to ambient temperature for 8 hours (No starting material by
TLC). The solution was concentrated to a yellow foam, THF ( 10 mL) was
added and the mixture was re-concentrated and used without further
purification.
Example 5-L
Synthesis of
3-Amino-1-methyl-4-phenyl-3,4-cis-dihydrocarbostyril
Step A: - Synthesis of 3-Amino-1-methyl-4-phenyl-3,4-trans-
dihydrocarbostyril
3-Amino-1-methyl-4-phenyl-3,4-traps-dihydrocarbostyril was prepared
following General Procedure 8-F using 3-azido-1-methyl-4-phenyl-3,4-trans-
dihydrocarbostyril from Example 5-K, Step C. The product was purified by
L.C. 2000 eluting with EtOAc/hexanes (4:1 ) to yield a white solid.
Selected 'H-NMR data for the title compound (CDC13): 8 = 4.03 (d, 1H,
J = 12.8 Hz), 3.92 (d, 1 H, J = 12.7 Hz).
Step B: - Synthesis of 3-(4-Chlorobenzylimine)-1-methyl-4-
phenyl-3,4-traps-dihydrocarbostyril
To a solution of the product from Step A (1 eq., 239.6 mg) in CH,CI,
( 10 mL) under NZ at ambient temperature was added 4-chlorobenzaldehyde ( 1.05
eq., 140 mg, Aldrich), Et3N (I.4 eq., 185 mL), and MgS04 (3.6 eq., 411 mg).
The resultant mixture was stirred at room temperature for 73 hours. The solids
were removed by filtration through a plug of Celite, rinsing with CH,CI,, and
the filtrate concentrated to provide 3-(4-chlorobenzyiimine)-1-methyl-4-phenyl-
3,4-traps-dihydrocarbostyril as a thick white foam.
Step C: - Synthesis of 3-Amino-1-methyl-4-phenyl-3,4-cis
dihydrocarbostyril
To a solution of diisopropylamine (1.05 eq., 0.132 mL) in THF (5 mL)
under N., at -78°C was added a solution of n-BuLi (1.05 eq., 0.588 mL
of a 1.6
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M solution in hexanes) and the result solution was stirred for 30 minutes. To
' this solution was added the product from Step B (1.0 eq., 336 mg) in THF (2
mL) via cannula. The solution was allowed to warm to 0°C, then quenched
with 1 M aq. HCl (3 mL) and allowed to warm to room temperature with
" 5 stirring overnight. The product was extracted into H,O and washed with
EtOAc
( 1 x), then the aqueous acid was basified with 1 M aq. K,C03 and the product
extracted into EtOAc. The EtOAc extract was dried over Na,S04, filtered, and
concentrated to give 3-amino-1-methyl-4-phenyl-3,4-cis-dihydrocarbostyril.
Selected 'H-NMR data for the title compound (CDC13): b = 4.31 (d, IH,
J = 6.6 Hz).
Example 5-M
Synthesis of
3-Amino-1-tert-butoxycarbonyl-4-phenyl
3,4-traps-dihydrocarbostyril/Tin Complex
Step A: - Synthesis of 1-tert-Butoxycarbonyl-4-phenyl-3,4-
dihydrocarbostyril
I -tent-Butoxycarbonyl-4-phenyl-3,4-dihydrocarbostyril was prepared from
the product of Example 5-K, Step A (CAS# 4888-33-9) by the Boc procedure
for aryl amides described by Grehn, L.; Gunnarsson, K.; Ragnarsson, U. Acta
Chemica Scandinavica B 1986, 40, 745-750; employing (Boc),O (Aldrich) and
catalytic DMAP (Aldrich) in acetonitrile. The product was purified by flash
chromatography eluting with CH,CI, gradient to CH,Ch/EtOAc ( I 9:1 ) and
isolated as a pale yellow oil.
Step B - Synthesis of 3-Azido-1-tert-butoxycarbonyl-4-phenyl-
3,4-traps-dihydrocarbostyril
Following General Procedure 8-K using the product from Step A, the
title compound was prepared as a 12.4:1 mixture of translcis isomers which
were separated by flash chromatography eluting with hexanes/Et,O (6:1 gradient
to 4:1 ) in the first column and hexanes/EtOAc ( I 2:1 ) in a second column.
The
pure traps isomer was used in Step C.
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Selected 'H-NMR data for the title compound (CDC13): 8 = 4.45 {d, 1H,
J = 11.1 Hz), 4.24 (d, 1 H, J = 11.2 Hz).
Step C: - Synthesis of 3-Amino-1-tert-butoxycarbanyl-4-phenyl-
3,4-traps-dihydrocarbostyril/Tin Complex
To a mixture of SnCI~ (450.6 mg) in MeOH {9 mL) under N, at 0°C was
added the product from Part D (433.0 mg) in MeOH ( I 5 mL) via cannula over a
period of 1 minute. The cooling bath was removed the solution allowed to
warm to ambient temperature for 17 hours. The solution was concentrated to an
amorphous yellow solid and used without further purification.
Example 5-N
Synthesis of
(S)-3-Amino-1-benzyl-b-valerolactam
Step A: - Synthesis of L-(+)-Ornithine Methyl Ester
Hydrochloride
Into a stirred suspension of I,-(+)-ornithine hydrochloride (Aldrich) in
methanol was bubbled anhydrous hydrochloric acid gas until the solution was
saturated. The reaction mixture was capped with a rubber septum and stirring
was continued overnight at room temperature. The solvent was then stripped
under reduced pressure and the residue triturated with ether. The resulting
solid
was dried under reduced pressure to afford L-(+)-ornithine methyl ester
hydrochloride as a white solid (97% yield).
Step B: - Synthesis of (S)-3-Amino-b-valerolactam
Sodium spheres in oil (2.0 eq.) (Aldrich) were washed with hexanes (2x)
and methanol (2.3 mL/mmol) was slowly added. The reaction mixture was
stirred under nitrogen until the sodium dissolved and then L-(+)-ornithine
methyl
ester hydrochloride ( 1 eq.) in methanol (2.3 mL/mmol) was added dropwise.
The reaction mixture was stirred for 16 hours and then diluted with diethyl
ether
{5 mL/mmol) and filtered to remove the solids. The solvent was then removed
under reduced pressure and the residue was heated at 70°C for 3 hours
under
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reduced pressure. The residue was then triturated with dichloromethane/ether,
' the solvent decanted and the resulting residue dried under reduced pressure
to
afford (S)-3-amino-8-valerolactam (44% yield).
' S Ste~C: - Synthesis of N-Boc-(S)-3-Amino-b-valerolactam
(S)-3-Amino-8-valerolactam (1 eq.) was dissolved in dioxane and the
solution was chilled to 0°C. BOC-anhydride (1.3 eq.) was added and the
ice
bath was removed allowing the solution to come to room temperature and
stirring was continued for 16 hours. The solution was rotory evaporated to
afford N-Boc-(S)-3-amino-b-valeroiactam.
Step D: - Synthesis of (S)-3-Amino-1-benzyl-b-vaierolactam
Following General Procedure 5-A and using N-Boc-(S)-3-amino-8-
valerolactam and benzyl bromide provided N-Boc-(S)-3-amino-1-benzyl-8-
valerolactam. Removal of the Boc group using General Procedure S-B afford
the title compuound.
Example 5-O
Synthesis of
4-Amino-2-aza-2-benzyl-3-oxobicyclo[3.2.1 )octane Hydrochloride
Ste~A: - Synthesis of 2-Aza-3-oxobicyclo[3.2.1]octane and 3-Aza-
2-oxobicyclo[3.2.1]octane (9:1 Mixture)
To (~)-norcamphor (Aldrich) in 1 mL/mmole of acetic acid was
added 1.5 eq. of hydroylamine-O-sulfonic acid. The reaction mixture was
heated to reflux under nitrogen for 1 hour and then saturated sodium carbonate
and dilute sodium hydroxide were added. The resulting mixture was extracted
with dichloromethane and the organic extracts washed with brine, dried over
sodium sulfate, and the solvent removed under reduced pressure. Purification
of
the residue by column chromatography afforded a 9:1 mixture of 2-aza-3-
oxobicyclo[3.2.1 ]octane and 3-aza-2-oxobicyclo[3.2.1 ]octane.
Stets B: - Synthesis of 2-Aza-2-benzyl-3-oxobicyclo[3.2.1]octane
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Following General Procedure S-A and using the product for Step A and
benzyl bromide, 2-aza-2-benzyl-3-oxobicyclo[3.2.1]octane was prepared.
Step C: - Synthesis of 2-Aza-2-benzyl-4-oximino-3-
S oxobicyclo[3.2.1]octane
To a solution of 2-aza-2-benzyl-3-oxobicyclo[3.2.1 ]octane in THF was
added 2.5 eq. of 1 M t-BuOKITHF (Aldrich) and the resulting mixture was
stirred for 30 minutes. Isoamyl nitrite (1.S eq.) was then added dropwise and
the reaction mixture was stirred overnight. To the reaction mixture was added
3N HCI and this mixture was extracted with ethyl acetate and the organic
extracts washed with water, dried, and concentrated under reduced pressure.
The residue was triturated with ether/hexanes, the solvents decanted and the
residue dried under reduced pressure to afford 2-aza-2-benzyl-4-oximino-3-
oxobicyclo[3.2.1 ]octane as a tan liquid (41 % yield). This procedure is
further
1S described in Y. Kim, Tetrahedron Lett. 30(21), 2833-2636 (1989}.
Step D: - Synthesis of 2-Aza-2-benzyl-4-amino-3-
oxobicyclo[3.2.1]octane
A solution of 2-aza-2-benzyl-4-oximino-3-oxobicyclo[3.2.1 ]octane in 10
mL/mmole of ethanol and S.8 mLlmmole of 3N HCl containing O.S g/mmole of
10% Pd/C was saturated with hydrogen gas to 45 psi. The mixture was shaken
for 3 hours and then filtered through a layer of Celite. The filtrate was
dried
over sodium sulfate and concentrated under reduced pressure to afford the
title
compound as a solid (86% yield). This procedure is further described in E.
Reimann, Arch. Pharm. 310, 102-109 (1977).
GENERAL PROCEDURE 6-A
Alkylation of
1-Amino- I ,3,4,5-tetrahydro-2H-3-benzazepin-2-one
Step A: 1-Ethoxycarbonylamino-1,3,4,5-tetrahydro-2H-3-benzazepin-2-
one was prepared according to the procedure of Ben-Ishai et al., Tetrahedron,
1987, =l3, 430.
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Step B: 1-Ethoxycarbonylamino-1,3,4,5-tetrahydro-2H-3-benzazepin-2-
one (2.0 g, 100 M%) was dissolved in DMF (30 mL) and NaH (95%, 0.17 g,
' 100M%) was added in one portion. The reaction mixture was stirred for 1 hour
and then the appropriate alkyl iodide (300M%) was added and the mixture was
' S stirred for 12 hours. The reaction was poured into water and extracted
with
ethyl acetate (3x). The ethyl acetate extracts were then washed with water
(3x)
and brine ( 1 x). Treatment with MgS04, rotoevaporation, and chromotography
(30% EtOAc/hexanes) yielded 1-ethoxycarbonylamino-3-alkyl-1,3,4,5-tetrahydro-
2H-3-benzazepin-2-one in 87% yield.
St- ep C: 1-Ethoxycarbonylamino-3-alkyl-1,3,4,5-tetrahydro-2H-3-
benzazepin-2-one ( 1.Og, 1 OOM%) was suspended in 30 mL of 30% HBr/HOAc
and heated to 100°C. The reaction mixture was stirred for 5 hours at
this
temperature and then the reaction was cooled and rotoevaporated to yield 1-
amino-3-alkyl-1,3,4,5-tetrahydro-2H-3-benzazepin-2-one as the hydrobromide
salt ( 100% yield).
GENERAL PROCEDURE 6-B
Alkylation of
3-Amino-1,3,4,5-tetrahvdro-2H-1-benzazepin-2-one
Step A: 3-Amino-1,3,4,5-tetrahydro-2H-1-benzazepin-2-one was
prepared from a-tetralone using the methods described in Armstrong et a!.
Tetrahedron Letters. 1994, 35, 3239. The following compounds were as
prepared by this procedure for use in the following steps:
5-methyl-3-amino-1,3,4,5-tetrahydro-ZH-1-benzazepin-2-one (from 4-
methyl-a-tetralone (Aldrich)); and
5 , 5-dimethyl-3-amino-1, 3 ,4 , 5-tetrahydro-2H- i -benzazepin-2-one (from
4,4-dimethyul-a-tetralone (Aldrich)).
Step B: 3-Amino-1,3,4,5-tetrahydro-2H-1-benzazepin-2-one (4.43 g,
100M % ) was suspended in t-butanol (30mL) and BOC-anhydride (7.5 mL,
130M % ) was added dropwise . The reaction was stirred for 2 hours and then it
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was rotoevaporated to a residue which was chromatographed with 60 % ethyl
acetatelhexanes to yield BOC-protected 3-amino-1, 3 , 4 , 5-tetrahydro-2H-1-
benzazepin-Z-one in 87 % yield.
St_ ep C: BOC-protected 3-amino-1,3,4,5-tetrahydro-2H-1-benzazepin-2-
one (1.5 g, 100M%) was dissolved in DMF (20mL) and NaH (95%, 0.13g,
100M%) was added in one portion. The reaction mixture was stirred for 1
hour and then the appropriate alkyl iodide (300M %) was added and stirring was
continued for 12 hours. The reaction was poured into water and extracted with
ethyl acetate (3x). The ethyl acetate extracts were washed with water (3x) and
then brine (lx). Treatment with MgS04, rotoevaporation, and chromotography
{30% EtOAc/hexanes) yielded a BOC-protected 3-amino-1-alkyl-1,3,4,5-
tetrahydro-2H-1-benzazepin-2-one in 80% yield.
Step D: The BOC-protected 3-amino-1-alkyl-1,3,4,5-tetrahydro-2H-1-
benzazepin-2-one (1.Og, 100M % ) was suspended in 30 mL of 1:1
CHZC121triflouroacetic acid and the mixture was stirred for 4 hours. The
reaction was then rotoevaporated to yield the 3-amino-1-alkyl-1,3,4,5-
tetrahydro-2H-1-benzazepin-2-one (100% yield).
Example 6-A
Synthesis of
3-Amino-1,5-dimethyl-1,3,4,5-tetrahydro-2H-1-benzazepin-2-one
Step A: 3-Amino-5-methyl-1,3,4,5-tetrahydro-2H-1-benzazepin-2-one
was prepared from 4-methyl-cx-tetralone using the methods described in
Armstrong et al. Tetrahedron Letters. 1994, 35, 3239.
Step B: 3-Amino-5-methyl-1,3,4,5-tetrahydro-2H-1-benzazepin-2-one
(9.3g 100M %) was dissolved in dioxane (300mL) and the solution was chilled
to 0°C. BOC-anhydride (13.89g 130M%) was added and the ice bath was
removed allowing the solution to come to room temperature and stirring was
continued for 16 hours. The solution was rotory evaporated to remove dioxane
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to provide an off white solid. This solid was .recrystallized from Cl-ICI3 to
' yield BOC-protected 3-amino-5-methyl-1,3,4,5-tetrahydro-2H-1-benzazepin-2-
one in 55 % -yield.
~ 5 Step C : BOC-protected 3-amino-5-methy 1-1, 3 , 4, 5-tetrahydro-2H-1-
benzazepin-2-one { 100 M % ) was dissolved in DMF (20mL) and NaH (95 % ,
100 M % ) was added in one portion and the reaction mixture was stirred for 1
hour. Methyl iodide (300 M %) was added and this mixture was stirred far 12
hours. The reaction was then poured into water and extracted with ethyl
acetate (3x) then backwashed with water (3 x) and then brine ( 1 x) .
Treatment
with MgS04, rotoevaporation) and chromotography (5 % MeOH/CH,CIz)
yielded BOC-protected 3-amino-1, 5-dimethyl-1, 3,4, S-tetrahydro-2H-1-
benzazepin-2-one in 75 % yield.
Step D: BOC-protected 3-amino-1,5-dimethyl-1,3,4,5-tetrahydro-2H-1-
benzazepin-2-one ( 100 M % ) was suspended in 30 mL of 1:1
CHZCI2/triflouroacetic acid. The reaction mixture was stirred for 4 hours. The
reaction was then rotoevaporated to yield 3-amino-1,5-dimethyl-1,3,4,5-
tetrahydro-2H-1-benzazepin-2-one (100% yield).
Example 6-B
Synthesis of
S-(L-Alaninyl)-amino-3,3,7-trimethyl
5,7-dihydro-6H-benz[b]azepin-6-one Hydrochloride
Following the procedure of Example 7-I and using 5-amino-3,3,7-
trimethyl-5,7-dihydro-6H-Benz[b)azepin-6-one hydrochloride (Example 6-C),
the title compound was prepared.
Example 6-C
Synthesis of
S-Amino-3,3, 7-trimethyl-S, 7-dihydro-
' 6H-benz[b]azepin-6-one Hydrochloride
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Step A: Following General Procedure .5-A and using N-t-Boc-5-amino-
3,3-dimethyl-5,7-dihydro-6H-Benz[b]azepin-6-one (General Procedure 6-B,
following bj~ Boc protection) and methyl iodide, N-t-Boc-5-amino-3 , 3, 7-
trimethyl-5,7-dihydro-6H-bent[b]azepin-6-one was prepared. -
Step B: Following General Procedure 8-N and using N-t-Boc-5-amino-
3,3,7-trimethyl-5,7-dihydro-6H-bent[b]azepin-6-one, the title compound was
prepared.
Example 6-D
Synthesis of
3-(S)-Amino-1-methyl-5-oxa-1,3,4,5
tetrahydro-2H-1-benzazepin-2-one
Ste~A: 3-(S)-Amino-5-oxa-1, 3,4, 5-tetrahydro-2H-1-benzazepin-2-one
was prepared from N-Boc-serine (Bachem) and 2-fluoro-1-nitrobenzene
(Aldrich) using the method of R. J. DeVita et al. , Bioorganic and Medicinal
Chemistry Lett. 1995, 5(12) 1281-1286.
Step B: Following General Procedure 5-A and using the product from
Step A, the title compound was prepared.
Example 6-E
Synthesis of
3-(S)-Amino-1-ethyl-5-oxa-1,3,4,5
tetrahydro-2H-1-benzazepin-2-one
Step A: 3-(S)-Amino-5-oxa-1, 3 , 4, 5-tetrahydro-2H-1-benzazepin-2-one
was prepared from N-Boc-serine (Bachem) and 2-fluoro-1-nitrobenzene
(Aldrich) using the method of R. J. DeVita et al. , Bioorganic and Medicinal
Chemistry Lett. 1995, 5(12) 1281-1286.
Step B: Following General Procedure 5-A and using the product from
Step A, the title compound was prepared.
T.
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Example 6-F _
Synthesis of
- 3-(S)-Amino-1-methyl-5-thia-1,3,4,5-
tetrahydro-2H-1-benzazepin-2-one
The title compound was prepared from N-Boc-cystine (Novabio) and 2-
' fluoro-1-nitrobenzene (Aldrich) using the method of R. J. DeVita et al. ,
Bioorganic and Medicinal Chemistry Lett. 1995, 5(12) 1281-1286, followed by
General Procedure 5-A.
GENERAL PROCEDURE 7-A
Preparation of
5-Amino-7-alkyl-5,7-dihydro
6H-dibenzlb,dlazepin-6-one Derivatives
Step A: Following General Procedure 5-A and using 5,7-dihydro-6H-
dibenz[b,d]azepin-6-one and an alkyl halide, the 7-alkyl-5,7-dihydro-6H-
dibenz[b,d]azepin-6-one was prepared.
Step B: The 7-alkyl-5,7-dihydro-6H-dibenz[b,d]azepin-6-one (1 eq.}
was dissolved in THF and isoamylnitrite ( 1.2 eq. ) was added. The mixture was
cooled to 0°C in an ice bath. NaHMDS (1.1 eq., 1M in THF) was added
dropwise. After stirring for 1 hour or until the reaction was complete, the
mixture was concentrated then acidified with 1N HCI and extracted with
EtOAc. The organic portion was dried and concentrated to yield a crude
product which was purified by silica gel chromatography.
Step C: The resulting oxime was dissolved in EtOHINH~ (20:1) and
hydrogenated in a bomb using Raney nickel and hydrogen (500 psi) at
100°C
for 10 hours. The resulting mixture was filtered and concentrated to provide
an
' oil which was purified by silica gel chromatography to yield the title
compound.
' 30
GENERAL PROCEDURE 7-B
' Preparation of
Fluoro-substituted 5,7-dihydro-6H-
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dibenzfb,dlaze~in-6-one Derivatives -
A modification of the procedure of Robin D. Clark and Jahangir,
Tetrahedron; Vol. 49, No. 7, pp. 1351-1356, 1993 was used. Specifically, an
appropriately substituted N-t-Boc-2-amino-2'-methylbiphenyl vas dissolved in
THF and cooled to -78 °C . s-Butyl lithium ( 1. 3M in cyclohexane, 2.2
eq. ) was
added slowly so that the temperature remained below -65 °C. The
resulting
mixture was allowed to warm to -25 ° C and was stirred at that
temperature for 1
hour. The mixture was cooled to -78°C. Dry COZ was bubbled through the
mixture for 30 seconds. The mixture was allowed to warm to ambient
temperature then was carefully quenched with water. The mixture was
concentrated under reduced pressure then was adjusted to pH 3 with 1 N HCI.
The mixture was extracted with EtOAc and the organic portion was dried and
concentrated to yield a crude material. The crude material was dissolved in
methanol and the solution was saturated with HCI. The mixture was heated at
reflux for 12 hours then was allowed to cool. The mixture was concentrated to
provide crude lactam which was purified by chromatography or crystallization.
GENERAL PROCEDURE 7-C
Resolution of
5-Amino-7-methyl-5 , 7-dihydro-6H-dibenz f b, dl azepin-6-one
In a round bottom flask was added the racemic freebase amine (1.0 eq.)
in methanol followed by di p-toluoyl-D-tartaric acid monohydrate ( 1. 0 eq . )
.
The mixture was concentrated in vacuo to~ a residue and redissolved in a
moderate volume of methanol and allowed to stir at room temperature open to
the atmosphere (8-72 hours). The solid was removed by filtration. The
enantiomeric excess was determined by chiral HPLC (Chiracel ODR) using
15 % acetonitrile and 85 % HZO with 0.1 % trifluoroacetic acid and a flow rate
of
1.0 mLlmin at 35°C. The resolved di p-toluoyl-D-tartaric salt was then
dissolved in EtOAc and saturated NaHC03 until pH 9-10 was reached. The
layers were separated and the organic layer was washed again with saturated
NaHC03, HZO, arid brine. The organic layer was dried over MgS04 and the
drying agent was removed by filtration. The filtrate was concentrated in
vacuo.
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The free amine was dissolved in MeOH and HCI (12M, 1.0 eq.) was added.
The salt was concentrated in vacuo and the resulting film was triturated with
EtOAc. Th$ HCI salt was filtered and rinsed with EtOAc. The ee was
determined by chiral HPLC.
r
Example 7-A
Synthesis of
5-Amino-7-methyl-5, 7-dihydro
6H-dibenz[b,d]azepin-G-one Hydrochloride
Step A - Synthesis of 7-Methyl-5,7-dihydro-6H-dibenzfb dlaze~in-6-one
A round bottom flask was charged with sodium hydride (0.295 g, 7.46
mmol) in 9.0 ml of DMF and treated with 5,7-dihydro-6H-dibenz[b,dJazepin-6-
one (1.3 g, 6.22 mmol) (CAS # 20011-90-9, prepared as described in Brown,
et. al. , Tetrahedron Letters, No. 8, 667-670, ( 1971 ) and references cited
therein) . After stirring at 60 ° C for 1 h, the solution was treated
with methyl
iodide (1.16 ml, 18.6 mmol) and stirring continued for 17 h with the exclusion
of light. After cooling, the reaction was diluted with CHZC12/H20, washed with
NaHS04 solution, HzO, and dried over NaZS04. Evaporation and flash
chromatography (SiOz, CHC13) gave 0.885 g (63 %) of the title compound as a
colorless solid.
NMR data was as follows:
'H-nmr (CDCl3): d = 7.62 (d, 2H), 7.26-7.47 (m, 6H), 3.51 (m, 2H),
3.32 (s, 3H).
C,SH,3N0 (MW = 223.27); mass spectroscopy (MH+) 223.
Anal. Calcd for C,SH,3N0; C, 80.69 H, 5.87 N, b.27. Found: C, 80.11
H, 5.95 N, 6.23.
Step B - Synthesis of 7-Methyl-5-oximo-~ 7-dihydro-6H-
dibenz[b,d~azepin-6-one
The compound isolated above (0.700 g, 3.14 mmol) was dissolved in 20
ml of toluene and treated with butyl nitrite (0.733 ml, 6.28 mmol). The
reaction
temperature was lowered to 0°C and the solution was treated with KHMDS
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(9.42 ml, 0.5 M) under N~ atmosphere. After -stirring for 1 h the reaction was
quenched with a saturated solution of NaHS04, diluted with CH,Ch and
separated. The organic layer was dried over Na~S04 and the title compound
purified by chromatography (SiO;, 98:2 CHC13/MeOH) giving 0.59 g (80 %) as
a colorless solid.
C,SH,,N,O, (MW = 252.275); mass spectroscopy {MH+) 252.
Anal. Calcd for C,SH,,N~O,; C, 71.42 H, 4.79 N, 11.10. Found: C, 71.24
H, 4.69 N, 10.87.
Step C - Synthesis of 5-Amino-7-Methyl-5,7-dihydro-6H-
dibenzL.dlazepin-6-one Hydrochloride
The oxime isolated above (0.99 g, 3.92 mmol) was hydrogenated in a
Parr apparatus at 35 psi over 10 % Pd/C (0.46 g) in 3A ethanol. After 32 h the
reaction mixture was filtered through a plug of celite, the filtrate
evaporated to a
foam and treated with a saturated solution of HCl (g) in Et20. The resulting
colorless solid was filtered, rinsed with cold Et~O and vacuum dried to give
0.66
g (61 %} of the title compound.
NMR data was as follows:
'H-nmr (DMSOd6): d = 9.11 (bs, 3H), 7.78-7.41(m, 8H), 4.83 (s, 1H),
3.25 (s, 3H).
C,SH,4N,0 HCl (MW = 274.753); mass spectroscopy (MH+ free base)
238.
Anal. Calcd for C,SH,qN,O HCI; G, 65.57 H, 5.50 N, 10.19 Found: C,
65.27 H, 5.67 N, 10.13.
Example 7-B
Synthesis of
(S)- and (R)-5-(L-Alaninyl)-amino-7-methyl-
5,7-dihydro-6H-dibenz[b,d]azepin-6-one
Step A - Synthesis of (S)- and (R)-5-(N-Boc-L-Alaninyl)-amino-7-
methvl-5 7-dihydro-6H-dibenz[b,d]azepin-6-one
Boc-L-Alanine (0.429 g, 2.26 mmol} (Aldrich) was dissolved in THF and
treated with HOBt hydrate (0.305 g, 2.26 mmol), and S-amino-7-methyl-5,7-
..___._..._.._. __... ...,.~-.....,....~..-.,..~,................ r.
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dihydro-6H-dibenz[b,d]azepin-6-one (0.45 g, 1:89 mmol) (Example 7-A). The
- temperature was lowered to 0°C and the reaction mixture treated with
EDC
(0.449 g, 2.26 mmol) (Alrich) and stirred 17 hours under N,. The reaction
mixture was evaporated, the residue diluted with EtOAc/H20, washed 1.0 N
- 5 HCI, sat. NaHC03, brine and dried over Na,S04. The diastereomers were
separated on a Chiralcel OD column using 10% IPA/heptane at 1.5 ml/minute.
Isomer 1: Retention time 3.37 minutes.
NMR data was as follows:
'H-nmr (CDC13): d = 7.62-7.33 (m, 9H), 5.26 (d, 1 H), 5.08 (m, I H),
4.34 (m, 1 H), 3.35 (s, 3H), 1.49 (s, 9H), 1.40 {d, 3H).
Optical Rotation: [a]~o = - 96 @ 589 nm (c = 1, MeOH).
C~3HZ,N30a (MW = 409.489); mass spectroscopy (MH+) 409.
Anal. Calcd for C,3H,,N3O4; C, 67.46 H, 6.64 N, 10.26. Found: C, 68.42
H, 7.02 N, 9.81.
Isomer 2: Retention time 6.08 minutes.
NMR data was as follows:
'H-nmr (CDC13): d = 7.74 (bd,1 H), 7.62-7.32 (m, 8H), 5.28 (d, 1H),
4.99 (m, 1H), 4.36 (m, 1H), 3.35 (s, 3H), 1.49 (s, 9H), 1.46 {d, 3H).
Optical Rotation: [a]~o = 69 @ 589 nm (c = 1, MeOH).
C~3H,,N304 (MW = 409.489); mass spectroscopy (MH+) 409.
Anal. Calcd for C,3H,,N3O4; C, 67.46 H, 6.64 N, 10.26. Found: C, 67.40
H, 6.62 N, 10.02
Step B - Synthesis of (S)- and (R)-~L-Alaninyl)-amino-7-methyl-
5 7-dihydro-6H-dibenz[b d,]azepin-6-one Hydrochloride
' The compounds isolated in Part A (each isomer separately) were
dissolved in dioxane and treated with excess HCl (g). After stirring for 17
hours, the title compounds were isolated as colorless solids after evaporation
and
' vacuum drying.
Isomer l:
C,gH,9N30=.HCI (MW = 345.832); mass spectroscopy (MH+ free base)
309.
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Optical Rotation: [a]ZO = - 55 @ 589 nm (c = 1, MeOH). -
Isomer 2:
C,8Ht9N30~.HC1 (MW = 345.832); mass spectroscopy (MH+ free base)
309. -
Optical Rotation: [a]ZO = 80 @ 589 nm (c = 1, MeOH).
Example 7-C
Synthesis of
(S}- and (R)-5-(L-Valinyl)-amino-7-methyl-
5,7-dihydro-6H-dibenz[b,d) azepin-6-one
Step A - Synthesis of (S)- and (R)-5-~N-Boc-L-Valinyl)-amino-7-
methyl-5,7-dihydro-6H-dibenz[b,d]azepin-6-one
Boc-L-Valine (0.656 g, 3.02 mmol) (Aldrich) was dissolved in THF and
treated with HOBt hydrate (0.408, 3.02 mmol), Dipea ( 1.05 ml, 6.05 mmol)
and 5-amino-7-methyl-5,7-dihydro-6H-dibenz[b,d]azepin-6-one hydrochloride
(0.75 g, 2.75 mmol)(Example 7-A). The temperature was lowered to 0°C
and
the reaction mixture treated with EDC (0.601 g, 3.02 mmol)(Alrich) and stirred
17 hours under N,. The reaction mixture was evaporated, the residue diluted
with EtOAc/H~O, washed 1.0 N HCI, sat. NaHC03, brine and dried over
Na,S04. The diastereomers were separated on a Chiralcel OD column using
10% IPA/heptane at 1.5 ml/minute.
Isomer 1: Retention time 3.23 minutes.
Optical Rotation: [a]ZO = - 120 @ 589 nm (c = 1, MeOH).
C~SH3,N3Oq (MW = 437.544); mass spectroscopy (MH+) 438
Isomer 2: Retention time 6.64 minutes.
Optical Rotation: [a]2° = 50 @ 589 nm (c = 1, MeOH).
C,SH3,N~04 (MW = 437.544); mass spectroscopy (MH+) 438
Step B - Synthesis of (S)- and (R)-5-(L-Valinyl)-amino-7-methyl-
5,7-dihydro-6H-dibenz[b,d]azepin-6-one Hydrochloride
The compounds isolated in Part A (each isomer separately) were
dissolved in dioxane and treated with excess HCl (g). After stirring for 17
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hours, the title compounds were isolated as colorless solids after evaporation
and
- vacuum drying.
Isomer I:
C,oH,3N30,.HC1 (MW = 373.88); mass spectroscopy (MH+ free base)
. 5 338.
Optical Rotation: [a)Z~ _ - 38 @ 589 nm (c = 1, MeOH).
Isomer 2:
C,oH,3N30,.HCl {MW = 373.88); mass spectroscopy (MH+ free base)
338.
Optical Rotation: [a],o = 97 @ 589 nm {c = 1, MeOH).
Example 7-D
Synthesis of
(S)- and (R)-5-(L-tert-Leucine)-amino-7-methyl-
5,7-dihydro-6H-dibenz[b,d]azepin-6-one
Step A - Synthesis of (S)- and (R)-5-(N-Boc-L-tert-Leucinyl)-
amino-7-methyl-5 7-dihydro-6H-dibenzfb dlazepin-6-one
Boc-L-tent-Leucine (0.698 g, 3.02 mmol) (Fluka) was dissolved in THF
and treated with HOBt hydrate (0.408, 3.02 mmol), Dipea ( 1.05 ml, 6.05
mmol} and 5-amino-7-methyl-5,7-dihydro-6H-dibenz[b,d)azepin-6-one
hydrochloride (0.75 g, 2.75 mmol){Example 7-A). The temperature was
lowered to 0°C and the reaction mixture treated with EDC (0.601 g, 3.02
mmol)
(Alrich) and stirred 17 hours under N~. The reaction mixture was evaporated,
the residue diluted with EtOAclH,O, washed 1.0 N HCI, sat. NaHC03, brine and
dried over Na,SO~. The diastereomers were separated on a Chiralcel OD
column using 10% IPA/heptane at I.5 ml/minute.
Isomer 1: Retention time 3.28minutes.
Optical Rotation: [a]zo = - 128 @ 589 nm (c = 1, MeOH).
C,6H;3N3O4 (MW = 451.571); mass spectroscopy (MH+) 452
Isomer 2: Retention time 5.52 minutes.
Optical Rotation: [a]ZO = 26 @ 589 nm {c = 1, MeOH).
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C,6H33N3O4 (MW = 45 I .571 ); mass spectroscopy (MH+) 452
Step B - Synthesis of ~S)- and (R)-5-(L-tent-Leucinyl)-amino-7-
methyl-5,7-dih~dro-6H-dibenz[b,d]azepin-6-one
Hydrochloride
The compounds isolated in Part A (each isomer separately) were
dissolved in dioxane and treated with excess HCl (g). After stirring for 17
hours, the title compounds were isolated as colorless solids after evaporation
and
vacuum drying.
Isomer l:
C,,Hz5N30,.HCl (MW = 387.9I); mass spectroscopy (MH+ free base)
352.
Optical Rotation: [a],o = - 34 @ 589 nm (c = 1, MeOH}.
Isomer 2:
C,,H~SN30,.HC1 (MW = 387.91); mass spectroscopy (MH+ free base}
352.
Optical Rotation: [a]~~ = 108 @ 589 nm (c = 1, MeOH).
Example 7-E
Synthesis of
5-(N-Boc-Amino)-5,7-dihydro-6H,7H-dibenz(b,d] azepin-6-one
Step A - Synthesis of 5-Iodo-5,7-dihydro-6H-dibenz[b,dlazepin-6-
one
A solution of 5,7-dihydro-6H-dibenz[b,d]azepin-6-one (1.0 g, 4.77 mmol)
(Example 7-A) and Et3N ( 2.66 ml, 19.12 mmol) were stirred for 5.0 minutes at
-15°C in CH~Ch and treated with TMSI (I.36 ml, 9.54 mmol). After
stirring for
15 minutes I, ( 1.81 g, 7.16 mmol) was added in a single portion and the
reaction allowed to warm to 5-10°C over 3 h. The reaction was quenched
with
sat. Na,SOj, diluted with CH,Ch and separated. The organics were washed with
Na,S03 and NaHS03 and dried over MgS04. After filtration, the organics were
concentrated to approximately 20 ml and diluted with an additional 20 ml of
hexanes. The title compound was isolated as a tan precipitate by filtration.
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Step B - Synthesis of 5-Azido-5,7-dihydro-6H-dibenz[b dlazepin-6-
- one
The -iodide isolate above was dissolved in DMF and treated with 1.2
equivalents of NaN3. After stirring I7 h at 23°C the mixture was
diluted with
_ 5 EtOAc/H~O, separated, washed with brine and dried over MgS04. The title
compound was triturated from hot EtOAc as a tan powder.
Step C - Synthesis of 5-(N-Boc-Amino)-5 7-dihydro-6H 7H-
dibenz[b,d]azepin-6-one
The azide was dissolved in THF/H~O and stirred at 23°C for 17 h in
the
presence of 3.0 equivalents of Ph3P. The reaction was diluted with 50
HOAc/toluene, separated, the aqueous layer extracted with toluene and
evaporated to an oily residue. This was taken to pH 7.0 by the addition of 1 N
NaOH, the resulting HOAc salt was collected and vacuum dried. Finally, the
compound was treated with Boc anhydride ( 1.05 equivalents) and Et3N (2.1
equivalents) in THF. After stirring for 5 h at 23 °C the reaction was
filtered and
the title compound isolated as a colorless powder.
Example 7-F
Synthesis of
5-Amino-7-(2-methylpropyl)-5,7-dihydro-
6H-dibenz[b,d]azepin-6-one Hydrochloride
Step A - Synthesis of 5-(N-Boc-Amino)-7-(2-methylpropyl)-5 7-
dihydro-6H-dibenz[b dlazepin-6-one
A solution of 5-(N-Boc-amino)-5,7-dihydro-6I-I-dibenz[b,dJazepin-
6-one (0.2g, 0.617 mmol) (Example 7-E) in DMF was treated with CsZC03 (0.22
g, 0.678 mmol) and warmed to 60°C. To the reaction mixture was added 1-
iodo-2-methylpropane (0.078 ml, 0.678 mmol) and stirring continued for 17 h.
After cooling to 23 °C the mixture was diluted with CHZCIz, washed
with
- 30 several portions of brine and dried over Na~SOa. The title compound was
purified by chromatography (SiO,, CHCI3/MeOH 9:1 ).
C23HZgN2O3 {MW = 380.41); mass spectroscopy {MH+) 381
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Anal. Calcd for C,3HZ8N~O3; C, 72.61 H, 7.42 N, 7.36. Found: C, 72.31
H, 7.64 N, 7.17.
Step B - Synthesis of 5-Amino-7-(2-methylpropyl)=5,7-dihydro-6H-
dibenz[b,d]azepin-6-one Hydrochloride
The compound isolated in Part A was deprotected in dioxane saturated
with gaseous HCI. The title compound was isolated as a slightly colored solid
after evaporation and vacuum drying.
Example 7-G
Synthesis of
5-Amino-7-(methoxyacetyl)-5,7-dihydro
6H-dibenz[b,d]azepin-6-one Hydrochloride
Step A- Synthesis of 5-(N-Boc-Amino)-7-(methoxyacetyl -
dihydro-6H-dibenz[b,d]azepin-6-one
A solution of 5-(N-Boc-amino)-5,7-dihydro-6H-dibenz[b,d]azepin-
6-one ( 1.03, 3.08 mmol) (Example 7-E) in DMF was treated with Cs,CO; ( 1.10
g, 3.39 mmol) and warmed to 60°C. To the reaction mixture was added
bromomethyl acetate (0.321 ml, 3.39 mmol) (Aldrich) and stirring continued for
17 h. After cooling to 23 °C the mixture was diluted with CH,CI,,
washed with
several portions of brine and dried over NaZS04. The title compound was
purified by chromatography (SiO~, CHCl3).
CZZH24N~O5 (MW = 396.44); mass spectroscopy (MH+) 397
Anal. Calcd for C"H24N,05; C, 66.65 H, 6.10 N, 7.07. Found: C, 66.28
H, 5.72 N, 6.50.
Step B - Synthesis of S-Amino-7-(methoxyacetyll-5,7-dihydro-6H-
dibenz[b,d]azepin-6-one Hydrochloride
The compound isolated in Part A was deprotected in dioxane saturated
with gaseous HC1. The title compound was isolated as a colorless solid after
evaporation and vacuum drying.
C"H,6N,03 HCl (MW = 332.78); mass spectroscopy (MH+ free base)
297.
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Example 7-H -
Synthesis of
-- 5-Amino-7-(3,3-dimethyi-2-butanonyl)-
5,7-dihydro-6H-dibenz[b,d]azepin-6-one Hydrochloride
Step A- Synthesis of 5-(N-Boc-Amino)-7-(3 3-dimet~l-butanonyl)-
S,7-dihydro-6H-dibenz[b,d]azepin-6-one
A solution of 5-(N-Boc-amino)-5,7-dihydro-6H-dibenz[b,d]azepin-6-one
(0.2 g, 0.617 mmol) (Example 7-E) in DMF was treated with Cs,C03 (0.3 g,
0.925 mmol) and warmed to 60°C. To the reaction mixture was added 1-
chloro-
3,3-dimethyl-2-butanone (0.096 ml, 0.74 mmol) {Aldrich) and stirring continued
for 17 h. After cooling to 23 °C, the mixture was diluted with CH,CI,,
washed
with several portions of brine and dried over Na~S04. The title compound was
isolated as a colorless solid.
C,SH3oN,04 (MW = 422.522); mass spectroscopy (MH+) 423
Step B - Synthesis of 5-Amino-7-(3,3-dimethyl-2-butanon 1
dihydro-6H-dibenz(b,d]azepin-6-one Hydrochloride
The compound isolated in Part A was deprotected in dioxane saturated
with gaseous HCI. The title compound was isolated as a colorless solid after
evaporation and vacuum drying.
Example 7-I
Synthesis of
L-Alaninyl-5-amino-7-methyl-5,7-dihydro
6H-dibenz[b,d]azepin-6-one Hydrochloride
Step A: Following General Procedure III-D and using N-t-Boc-L-alanine
and 5-amino-7-methyl-5,7-dihydro-6H-dibenz[b,d]azepin-6-one, N-t-Boc-L-
alaninyl-5-amino-7-methyl-5,7-dihydro-6H-dibenz[b,d]azepin-6-one was
prepared.
Step B: Following General Procedure 8-N and using the N-t-Boc-L-
alaninyl-5-amino-7-methyl-5,7-dihydro-6H-dibenz[b,d]azepin-6-one, the title
compound was prepared. Other substituted N-t-Boc-L-alaninyl-5-amino-7-
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methyl-5,7-dihydro-6H-dibenz[b,d]azepin-6-ones can also be prepared by this
procedure.
Example 7-J
Synthesis of
L-Valinyl-5-amino-7-methyl-5,7-dihydro
6H-dibenz[b,d]azepin-6-one Hydrochloride
Step A: Following General Procedure III-D and using N-t-Boc-L-valine
and 5-amino-7-methyl-5,7-dihydro-6H-dibenz[b,d]azepin-6-one, N-t-Boc-L-
valinyl-5-amino-7-methyl-5,7-dihydro-6H-dibenz[b,d]azepin-6-one was prepared.
Step B: Following General Procedure 8-N and using the N-t-Boc-L-
valinyl-5-amino-7-methyl-5,7-dihydro-6H-dibenz[b,d]azepin-6-one, the title
compound was prepared. Other substituted N-t-Boc-L-valinyl-5-amino-7-
methyl-5,7-dihydro-6H-dibenz[b,d]azepin-6-ones can also be prepared by this
procedure.
Example 7-K
Synthesis of
5-Amino-7-phenbutyl-5,7-dihydro-6H-dibenz[b,d] azepin-6-one
Following General Procedure 7-A and using 5,7-dihydro-6H-
dibenz[b,d]azepin-6-one (prepared as described in Brown, et. al., Tetrahedron
Letters, No. 8, 667-670, ( 1971 ) and references cited therein} and 1-chloro-4-
phenylbutane (Aldrich), the title compound was prepared.
Example 7-L
Synthesis of
5-Amino-7-cyclopropymethyl-5,7-dihydro
6H-dibenz[b,d]azepin-6-one
Following General Procedure 7-A and using 5,7-dihydro-6H-
dibenz[b,d]azepin-6-one (prepared as described in Brown, et. al., Tetrahedron
Letters, No. 8, 667-670, ( 1971 ) and references cited therein) and
(bromomethyl)cyclopropane (Aldrich), the title compound was prepared.
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Example 7-M -
Synthesis of
~ ~ _ 5-Amino-7-(2',2',2'-trifluoroethyl)-5,7-dihydro-
6H-dibenz(b,djazepin-6-one
Following General Procedure 7-A and using 5,7-dihydro-6H-
dibenz[b,d]azepin-6-one (prepared as described in Brown, et. aL, Tetrahedron
Letters, No. 8, 667-670, ( 1971 ) and references cited therein) and 1-bromo-
2,2,2-
trifluoroethane (Aldrich), the title compound was prepared.
Example 7-N
Synthesis of
5-Amino-7-cyclohexyl-5,7-dihydro-
6H-dibenz[b,d)azepin-6-one
Following General Procedure 7-A and using 5,7-dihydro-6H-
dibenz[b,d]azepin-6-one (prepared as described in Brown, et. al., Tetrahedron
Letters, No. 8, 667-670, ( 1971 ) and references cited therein) and
bromocyclohexane (Aldrich), the title compound was prepared.
Example 7-O
Synthesis of
5-{L-Alaninyt)amino-9-fluoro-7-methyl
5,7-dihydro-6H-dibenz[b,d[azepin-6-one Hydrochloride
Step l: 2-Bromo-5-fluorotoluene was stirred in THF at -78C. s-BuLi
( 1.05 eq., 1.3 M in cyclohexane) was slowly added and the mixture was stirred
for 45 minutes. Trimethyiborate ( 1.5 eq) was added and the mixture was
allowed to warm to ambient temperature. After stirring for 1 hour, pinacol (2
eq.) was added. The mixture was stirred for 16 hours then was concentrated
under reduced pressure. 'i~he resulting residue was slurried in CH,Ch and
. filtered through Celite. The filtrate was concentrated to yield an oil which
was
purified by chromatography on deactivated silica gel (Et;N) to yield the
arylboronate ester.
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Step 2: 2-Bromoaniline ( 1 eq.) and di-t-butyl-dicarbonate ( 1-.1 eq:) were
stirred at 80°C for 20 hours. The resulting mixture was allowed to cool
and was
directly distilled using house vacuum to provide N-t-Boc-2-bromoaniline.
Step 3: N-t-Boc-2-bromoaniline (Step 2, 1 eq.), the arylboronate ester
(Step I, I.1 eq.), K,C03 (1.1 eq.) and
tetrakis(triphenylphosphine)palladium(0)
(0.02 eq) were stirred in 20% water/dioxane under nitrogen. The solution was
heated at reflux for 10 hours. The mixture was allowed to cool then was
concentrated. The resulting residue was partitioned between water and
chloroform. The organic portion was dried and concentrated to yield an oil
which was purified by silica gel chromatography using 1:1 CH,CI,/hexanes.
Step 4: Following General Procedure 7-B and using the substituted
biphenyl from step 3, the 9-fluoro-5,7-dihydro-6H-dibenz[b,d]azepin-6-one was
prepared.
Step 5: 9-Fluoro-5,7-dihydro-6H-dibenz[b,d]azepin-6-one ( 1 eq., Step 4},
cesium carbonate ( 1.1 eq.> Aldrich) and methyl iodide ( 1.1 eq., Aldrich)
were
stirred in dry DMF at ambient temperature for 16 hours. The mixture was
concentrated under reduced pressure to provide a residue which was partitioned
between EtOAc and water. The organic portion was dried and concentrated to
yield an oil which was purified by silica gel chromatography to 9-fluoro-7-
methyl-5,7-dihydro-6H-dibenz[b,d]azepin-6-one.
St. ep 6: Following General Procedure 7-A, Step B and 9-fluoro-7-
methyl-5,7-dihydro-6H-dibenz[b,d]azepin-6-one from Step 5, 5-amino-9-fluoro-
7-methyl-5,7-dihydro-6H-dibenz[b,d]azepin-6-one was prepared.
St, e~ 7: Following the procedure of Example 7-I and using S-amino-9
fluoro-7-methyl-5,7-dihydro-6H-dibenz[b,d]azepin-6-one from Step 6, the title
compound was prepared.
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Example 7-P -
Synthesis of
- ~ - 5-(L-Alaninyl)amino-13-fiuoro-7-methyl-
5,7-dihydro-6H-dibenz[b,d]azepin-6-one Hydrochloride
Following the procedure of Example 7-O and using 2-bromo-4-
fluoroaniline (Step 2, Lancaster) and o-toIylboronic acid (Step 3, Aldrich),
the
title compound was prepared.
Example 7-Q
Synthesis of
5-(L-Alaninyl)amino-IO-fluoro-7-methyl
5,7-dihydro-6H-dibenz[b,dJazepin-6-one Hydrochloride
Following the procedure of Example 7-O and using 2-bromo-4-
fluorotoluene (Step 1 ), the title compound was prepared.
Example 7-R
Synthesis of
5-(L-Alanyl)-amino-7-cyclopropylmethyl
5,7-dihydro-6H-dibenz[b,d]azepin-6-one Hydrochloride
Following the procedure of Example 7-I and using 5-amino-7-
cyclopropylmethyl-5,7-dihydro-6H-dibenz[b,d]azepin-6-one (Example 7-L), the
title compound was prepared.
Example 7-S
Synthesis of
5-(L-Alaninyl)amino-7-phenbutyl
5,7-dihydro-6H-dibenz(b,d]azepin-6-one Hydrochloride
Following the procedure of Example 7-1 and using 5-amino-7-phenbutyl-
5,7-dihydro-6H-dibenz[b,dJazepin-6-one (Example 7-K), the title compound was
prepared.
Example 7-T
Synthesis of
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5-(L-Valinyl)amino-7-cyclopropylmethyl- -
5,7-dihydro-6H-dibenz[b,d]azepin-6-one Hydrochloride
Following the procedure of Example 7-J and using 5-amino-7-
cyclopropylmethyl-5,7-dihydro-6H-dibenz[b,d]azepin-6-one (Example 7-L), the
title compound was prepared.
Example 7-U
Synthesis of
5-(L-Valinyl)amino-7-phenbutyl
5,7-dihydro-6H-dibenz[b,d]azepin-6-one Hydrochloride
Following the procedure of Example 7-J and using 5-amino-7-phenbutyl-
5,7-dihydro-6H-dibenz[b,d]azepin-6-one (Example 7-U), the title compound was
prepared.
Example 7-V
Synthesis of
5-(L-Valinyl)amino-7-hexyl-5,7-dihydro
6H-dibenz[b,d]azepin-6-one Hydrochloride
Step A: Following General Procedure 7-A and using 5,7-dihydro-6H-
dibenz[b,d]azepin-6-one (prepared as described in Brown, et. al., Tetrahedron
Letters, No. 8, 667-670, ( 1971 ) and references cited therein) and I -
bromohexane
(Aldrich}, 5-amino-7-hexyl-5,7-dihydro-6H-dibenz[b,d]azepin-6-one was
prepared.
St-ep B: Following the procedure of Example 7-J and using 5-amino-7-
hexyl-5,7-dihydro-6H-dibenz[b,d]azepin-6-one, the title compound was prepared.
Example 7-W
Synthesis of
5-(L-Valinyl)amino-10-fluoro-7-methyl-
5,7-dihydro-6H-dibenz[b,d]azepin-6-one Hydrochloride
Following the procedure of Example 7-J and using 5-amino-10-fluoro-7-
methyl-5,7-dihydro-6H-dibenz[b,d]azepin-6-one (as prepared in Examplel~-'~Q},
the title compound was prepared.
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Example 7-X
Synthesis of
-- 5-(L-Valinyl)amino-13-fluoro-7-methyl-
5,7-dihydro-6H-dibenz[b,dJazepin-6-one Hydrochloride
Following the procedure of Example 7-J and using the 5-amino-13-
fluoro-7-methyl-5,7-dihydro-6H-dibenz[b,d]azepin-6-one (as prepared in
Example 7-P), the title compound was prepared.
Example 7-Y
Synthesis of
5-(L-Valinyl)amino-13-fluoro-7-methyl-
5,7-dihydro-6H-dibenz~b,d]azepin-6-one Hydrochloride
Following the procedure of Example 7-J and using the 5-amino-9-fluoro-
7-methyl-5,7-dihydro-6H-dibenz[b,d]azepin-6-one (as prepared in Example 7-O),
the title compound was prepared.
Example 7-Z
Synthesis of
(5-Amino-7-methyl-1,2,3,4,5,7-hexahydro
6H-dicyclohexyl[b,d] azepin-6-one
The 5-amino-7-methyl-5,7-dihydro-6H-dibenz[b,d]azepin-6-one
hydrochloride (Example 7-A) was dissolved in a l:l mixture of EtOAc/HOAc.
5% Rh/C was added and the mixture was stirred at 60°C under 60 psi of
hydrogen. After 3 days, the mixture was filtered and the filtrate was
concentrated to provide an oil which was purified by SCX-ration exchange
chromatography to yield the title compound.
Example 7-AA
Synthesis of
S-(S)-Amino-7-methyl-5,7-dihydro-
6H-dibenz[b,d]azepin-6-one Hydrochloride
Following General Procedure 7-C using racemic 5-amino-7-methyl-5,7-
dihydro-6H-dibenz[b,d]azepin-6-one ( I .0 eq.) and di p-toluoyl-D-tartaric
acid
monohydrate ( 1,0 eq.) in methanol, the title compound was prepared as a
solid.
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The product was collected by filtration. Enantiomeric excess was determined by
chiral HPLC.
Desired enantiomer 1: retention time of 9.97 minutes.
Undesired enantiomer 2: retention time of 8.62 minutes.
NMR data was as follows:
'H-nmr (CDC13): 8 = 9.39 (s, 2H), 7.75-7.42 (m, 8H), 4.80 (s, 1 H), 3.30
{s, 3H).
C,SH,SCIN,O {MW = 274.75}; mass spectroscopy (MH+) 239.1.
Anal Calcd for C,SH,SC1N,03; C, 65.57; H, 5.50; N, 10.20; Found: C,
65 . S I , H, 5.61; N, I 0.01.
GENERAL PROCEDURE 8-A
N-1-Methylation of Benzodiazepines
A solution of benzodiazepine (1 eq.) in DMF (0.1 M concentration) at
0°C was treated with potassium tent-butoxide ( 1.0 eq., 1.0 M solution
in THF)
After stirring for 30 minutes at 0°C, iodomethane {1.3 eq.) was
added and
stirring continued for 25 minutes. The mixture was diluted with methylene
chloride and washed with water and brine. The organic phase was dried over
Na,S04, filtered, and concentrated. The crude product was then either purified
by trituration with l:l ether/hexanes or chromatographed via HPLC using ethyl
acetate/hexanes as the eluent.
GENERAL PROCEDURE 8-B
Cbz Removal Procedure
A flask was charged with the Cbz-protected 3-aminobenzodiazepine (1
eq.). To this was added HBr (34 eq.; 30% solution in acetic acid). Within 20
minutes all of the starting material dissolves. The reaction was stirred for 5
hours at ambient temperature. Ether was added to the orange solution causing
the HBr~amine salt to precipitate. The mixture was decanted. This process of
adding ether and decanting was repeated thrice in an effort to remove acetic
acid
and benzyl bromide. Toluene was added and the mixture concentrated in vacuo.
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This step was also repeated. The HBr salt was partitioned between -ethyl -
acetate
and 1 M K~C03. The aqueous layer was back-extracted with ethyl acetate. The
combined oiganics were washed with brine, dried over NaZS04, filtered, and
concentrated. -
- 5
GENERAL PROCEDURE 8-C
Boc Removal Procedure
A solution of Boc-protected amine (1 eq.) in methylene chloride (0.15 M
concentration} was cooled to 0°C and treated with trifluoroacetic acid
(30 eq.).
After 10 minutes at 0°C, the cooling bath was removed and stirring
continued at
ambient for 20 minutes to 1 hour. The mixture was concentrated in vacuo to
remove excess trifluoroacetic acid. The residue was dissolved in methylene
chloride and washed with saturated aqueous NaHC03 or 1 M K~C03 and brine.
The organic layer was dried over Na,SO~, filtered, and concentrated.
GENERAL PROCEDURE 8-D
Azide Transfer Reaction Using_KHMDS
The azido derivative was prepared using the procedure described in John
W. Butcher et al., Tet. Lett., 37, 6685-6688 (1996).
GENERAL PROCEDURE 8-E
Azide Transfer Reaction Using LDA
To a solution of diisopropylamine (1.1 eq.) in 1 mL of dry THF cooled
to -78°C was added n-butyl lithium {1.6M in hexane) (l.l eq.) dropwise
maintaing the reaction temperature at -78°C. The reaction mixture was
stirred
for 30 min. at -78°C and then the lactam (0.471 mM) was added dropwise
as a
solution in 1 mL of dry THF. The reaction mixture was stirred at -78°C
for 30
min. and then a pre-cooled solution of trisyl azide ( 1.2 eq.) was added as a
solution in 1 mL of dry THF. The reaction mixture was stirred at -78°C
for 20
min. and then quenched with acetic acid (4.0 eq. ). The reaction mixture was
then stirred at 40°C for 2 hrs. The reaction was then poured into EtOAc
and
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__ 278 __
washed with water, sodium bicarbonate and brine, and then dried over sodium
sulfate, filtered and concentrated. The residue was purified by LC 2000
chromatography.
GENERAL PROCEDURE 8-F
Azido Group Reduction
The azido group was reduced to the corresponding primary amine using
the procedure described in John W. Butcher et al., Tet. Lett., 37, 6685-6688
{ 1996).
GENERAL PROCEDURE 8-G
N-Alkylation of Amides or Lactams
Using Sodium Hydride or Potassium tent-Butoxide
To a slurry of sodium hydride or potassium tent-butoxide ( 1.1 eq) in 15
mL of dry DMF was added the appropriate amide (0.0042 moles) as a solution
in 10 mL of DMF. The alkyl iodide was then added and a thick slurry resulted.
The reaction became less thick as time elapsed and when complete by TLC the
reaction had become homogeneous. The reaction mixture was poured over ice
and extracted into ethyl acetate. The organic layer was washed with water,
followed by brine. The organic layer was then dried over sodium sulfate,
filtered and concentrated under reduced pressure. The residue was purified by
HPLC (LC 2000), eluting with an ethyl acetate/hexane system.
GENERAL PROCEDURE 8-H
N-Alkylation of
Amides or Lactams Using KHMDS
To the appropriate amide or lactam in THF cooled to -78°C was
added
KHMDS dropwise and the reaction mixture was stirred for 30 min. at -
78°C.
The alkyl iodide was then added dropwise while maintaining the temperature at
-70°C. The cooling bath was then removed and reaction was allowed to
warm
to room temperature and stirring was continued for 2 hours. The reaction
mixture was they poured over ice and extracted into ethyl acetate. The organic
r
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extracts were washed with water, followed by brine. The organic layer was then
dried over sodium sulfate, filtered and concentrated under reduced pressure.
The
residue was purified by HPLC {LC 2000), eluting with an ethyl acetate/hexane
system.
GENERAL PROCEDURE 8-I
N-Alkylation of Amides or Lactams Using Cesium Carbonate
To a solution of the amide or lactam in DMF was added cesium
carbonate ( 1.05 eq} and an alkyl iodide ( 1.1 eq). The mixture was allowed to
stir overnight at room temperature and then the reaction mixture was dilluted
with ethyl acetate and washed with water, followed by brine. The organic layer
was dried over sodium sulfate, filtered and concentrated under reduced
pressure.
The residue was purif ed by HPLC (LC 2000), eluting with an ethyl
acetate/hexane system.
GENERAL PROCEDURE 8-J
BOC Removal Procedure
To an N-Boc protected compound was added CH~CI~/TFA (4:1 ) at room
temperature. The reaction mixture was stirred at room temperature for 3 hours
and then concentrated. The residue was extracted into dichloromethane and
washed with water, saturated sodium bicarbonate, dried over Na~S04, filtered
and concentrated to give the free amine.
GENERAL PROCEDURE 8-K
Azide Transfer Procedure
This azide transfer procedure is a modification of the procedure described
in Evans, D. A.; Britton, T. C.; Ellman, J. A.; Dorow, R. L. J. Am. Chem. Soc.
1990, 112, 40 I I -4030. To a solution of the lactam substrate ( 1.0 eq. ) in
THF
(~0.1 M) under N, at -78 °C was added a solution of KN(TMS)~ (1.1 eq.
of 0.~
M in Toluene, Aldrich) dropwise over a period of 2-10 'minutes. A slight
exotherm was often observed by an internal thermometer, and the resulting
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solution was stirred for 5-15 minutes, while re-cooling to -78°C. Then,
trisyl
azide (1.1-1.5 eq., CAS No. 36982-84-0, prepared as described by references in
the Evans reference above) in THF (~0.5 M), either precooled to -78°C
or at
room temperature, was added via cannula over a period of 0.5-~ minutes.
Again, a slight exotherm was generally noted. The resulting solution was
stirred
for from 5-10 minutes, while re-cooling to -78°C. Then, AcOH (4. S-4.6
eq.,
glacial) was added, the cooling bath removed and the mixture allowed to warm
to room temperature with stirring for 12-16 hours. The mixture was diluted
with EtOAc, in a 2-5 volume multiple of the initial THF volume, and washed
with dilute aq. NaHC03 ( 1-2x), 0.1-1.0 M aq. HCl (0-2x}, and brine ( 1 x).
The
organic phase was then dried over MgSO~, filtered, concentrated to provide the
crude product.
GENERAL PROCEDURE 8-L
Azide Reduction to an Amine
A mixture of the azide in absolute EtOH (0.03-0.07 M) and 10% Pd/C
(~ 1 /3 by weight of the azide) was shaken in a Parr apparatus under H, {35-45
psi) at room temperature for 3-6 hours. The catalyst was removed by filtration
through a plug of Celite, rinsing with absolute EtOH, and the filtrate
concentrated to provide the crude amine product.
GENERAL PROCEDURE 8-M
Amide Alkylation Using Cesium Carbonate
This procedure is a modification of the procedure described in Claremon,
D. A.; et al, PCT Application: WO 96-US8400 960603. To a mixture of 2,4-
dioxo-2,3,4,5-tetrahydro-1H-1,5-benzodiazepine (CAS No. 49799-48-6) in DMF
( 1.0 eq., 0.7 M) under N~ at room temperature was added Cs~C03 (2.2 eq.) and
the appropriate alkyl halide (2.2 eq.). The mixture was stirred at room
temperature for 5. 5-16 hours. The mixture was partitioned between EtOAc and
sat. NaHC03. The aqueous layer was extracted with EtOAc ( 1-2x) and the
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combined EtOAc extracts were dried over Na,SOa, filtered, and concentrated to
provide the crude product.
GENERAL PROCEDURE 8-N
BOC Removal Procedure
A stream of anhydrous HCl gas was passed through a stirred solution of
the N-t-Boc protected amino acid in 1,4-dioxane (0.03-0.09 M}, chilled in a
ice
bath to ~10°C under NZ, for 10-15 minutes. The solution was capped, the
cooling bath removed, and the solution was allowed to warm to room
temperature with stirring for 2-8 hours, monitoring by TLC for the consumption
. of starting material. The solution was concentrated (and in some instances
dissolved in CH,C1, then re-concentrated and placed in vacuum oven at 60-
70°C
to remove most of the residual dioxane) and used without further purification.
Example 8-A
Synthesis of
3-Amino-1,3-dihydro-5-(I-piperidinyl)-2H-1,4-benzodiazepin-2-one
Step A - Preparation of 1 2-Dihydro-3H-1-methvl-5-(I-piperidinyl)-
1,4-benzodiazepin-2-one
A solution of phosphorous pentachloride ( 1.2 eq) in methylene chloride
was added dropwise to a solution of 1-methyl-1,2,3,4-tetrahydro-3H-1,4-
benzodiazepin-2,5-dione {Showell, G. A.; Bourrain, S.; Neduvelil, J. G.;
Fletcher, S. R.; Baker, R.; Watt, A. P.; Fletcher, A. E.; Freedman, S. B.;
Kemp,
J. A.; Marshall, G. R.; Patel, S.; Smith, A. J.; Matassa, V. G. J. Med. Chem.
1994, 37, 719.) in methylene chloride. The resultant yellowish-orange solution
was stirred at ambient temperature for 2.5 hours; the solvent was removed in
vacuo. The orange residue was redissolved in methylene chloride, cooled to 0
' °C, and treated with a solution of piperidine (2 eq) and
triethylamine (2 eq) in
methylene chloride. The cooling bath was removed and the reaction stirred for
18 hours. The reaction mixture was washed with saturated aqueous NaHC03
(back-extracted with methylene chloride) and brine. The organic phase was
dried over Na.,S04, filtered, and concentrated. The residue was purified via
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HPLC eluting with a gradient of 4 to 10% methanol/methylene chloride
affording the title intermediate as a yellow solid having a melting point of
103-
105°C. '
C,SH,9N30 (MW 257.37); mass spectroscopy 257.
S Anal. Calcd for C,SH,9N30: C, 70.01; H, 7.44; N, 16.33. Found: C,
69.94; H, 7.58; N, 16.23.
Step B - Preparation of 1,2-Dih~dro-3H-1-methyl-3-oximido-5-(1-
~peridinyl)-1,4-benzodiazepin-2-one
Potassium tert-butoxide (2.5 eq} was added in two portions to a -
20°C
solution of I ,2-dihydro-3 H-1-methyl-5-( 1-piperidinyl}-1,4-benzodiazepin-2-
one
( 1 eq) in toluene}. After stirring at - 20°C for 20 min, isoamyl
nitrite ( I .2 eq.;
Aldrich) was added to the red reaction mixture. The reaction was stirred at -
20
°C for 5 hours at which time the reaction was done by TLC. The cooling
bath
was removed and the reaction quenched with 0.5 M citric acid. After stirring
for 10 minutes, diethyl ether was added. The suspension was stirred at ambient
temperature overnight then filtered washing with ether. The resultant cream
colored solid had a melting point of 197-200°C.
'H NMR data of the E/Z isomers was as follows:
'H NMR (300 MHz, CDC13): 8 = 7.64 ( 1 H, bs), 7.48 (2H, d, J=7.4 Hz),
7.35-7.20 (6H, m), 6.75 (1H, bs), 3.8-3.2 (8H, m), 3.46 (3H, s), 3.42 (3H, s),
1.90-1.40 (12H, m).
C,SH,8N40z (MW = 286.37); mass spectroscopy 286.
Step C - Preparation of 1 2-dihydro-3H-I-methyl-3-[O-
(ethylaminocarbonyl)oximido]-5-( 1-piperidinyl)-1,4-
benzodiazepin-2-one
A mixture of 1,2-dihydro-3H-1-methyl-3-oximido-5-(1-piperidinyl)-1,4-
benzodiazepin-2-one ( I eq) in THF was treated with ethyl isocyanate ( 1.7 eq)
and triethylamine (0.6 eq). The mixture was heated to 64°C for 4 hours.
The
mixture was concentrated and the residue purified by HPLC eluting with 5%
methanollmethylene chloride.
_.................., . ,," ...
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~H NMR data of the E/Z isomers was as follows: _
'H NMR (300 MHz, CDC13): 8 = 7.50 (2H, dd, J=8.4, 1.5 Hz), 7.35-7.22
- (6H, m), 6.42 ( 1 H, bt), 6.20 ( I H, bt), 3.7-3.4 (8H, m), 3.46 (3H, s),
3.44 (3H,
s), 3.25 (4H, m), 1.9-1.4 ( 12H, rrl), I .12 (3H, t, J=6.3 Hz), 1.10 (3H, t,
J=6.3
Hz).
C,$H23NSO3 (MW = 357.46); mass spectroscopy 357.
Step D - Preparation of 3-Amino-1 3-dihydro-2H-1-methyl-5~1-
piperidinyl)-1,4-benzodiazepin-2-one
The I,2-dihydro-3H- I -methyl-3-[O-(ethylaminocarbonyl)oximido]-5-( 1-
piperidinyl)-I,4-benzodiazepin-2-one (I eq) was hydrogenated in methanol over
5% palladium on carbon (0.15 eq) at 43 psi for 3.25 hours. The reaction was
filtered through celite and concentrated irt vacuo. The residue was taken up
in
methylene chloride and filtered a second time through celite. The filtrate was
concentrated and the resultant foam was used immediately.
Example 8-B
Synthesis of
3-(L-Alaninyl)-amino-2,3-dihydro
1-m ethyl-5-phenyl-1 H-1,4-benzodiazepin-2-one
Step A - Preparation of (S)-3-amino-I 3-dihydro-1-meth=phe~l-
2H-I.4-benzodiazepin-2-one (1S)-7 7-dimethyl-2-
oxobicycloj2.2.1 ]heptane-1-methanesulfonate
The title intermediate was prepared. according to Reider, P. J.; Davis, P.;
Hughes, D. L.; Grabowski, E. J. J. .I. Org. Chem. 1987, 52, 955 using 3-amino-
1,3-dihydro-1-methyl-5-phenyl-2H-1,4-benzodiazepin-2-one (Bock M. G.;
DiPardo, R. M.; Evans, B. E.; Rittle, K. E.; Veber, D. F.; Freidinger, R. M.;
Hirshfield, J.; Springer, J. P. J. Org. Chem, 1987, 52, 3232.) as the starting
material.
Step B - Preparation of 3-fN'-(tert-Butylcarbamate)-L-alaninyll-
amino-2,3-dihydro-1-methyl-5-phenyl- I H- I 4-
benzodiazepin-2-one
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(S)-3-Amino-1,3-dihydro-I-methyl-5-phenyl-2H-1,4-benzodiazepin-2-one,
(1S)-7,7-dimethyl-2-oxobicyclo[2.2.1]heptane-1-methanesulfonate was free based
by partitioning between methylene chloride and 1 M potassium carbonate. The
free amine was then coupled with N-Boc-alanine following General Procedure
III-D.
C~~Hz$N404 (MW = 436.56); mass spectroscopy 436.
Anal. Calc. for C,4H.,8N4O4: C, 66.03; H, 6.47; N, 12.84. Found: C,
65.79; H, 6.68; N, 12.80.
Step C - Preparation of 3-f L-Alaninyl)-amino-2,3-dihydro- I -methyl-
5-phenyl-1H-1,4-benzodiazepin-2-one
Following General Procedure 8-C using 3-[N'-(tert-butylcarbamate)-L-
alaninyl]-amino-2,3-dihydro-1-methyl-5-phenyl-1 H- I ,4-benzodiazepin-2-one,
the
title compound was prepared as a white foam.
Anal. Calc. for C~9H~9N4O,: C, 69.21; H, 6.64; N, 15.37. Found: C,
70.11; H, 6.85; N, 15.01.
Example 8-C
Synthesis of
3-(L-Alaninyl)-amino-7-chloro-2,3-dihydro-
1-methyl-5-phenyl-1H-1,4-benzodiazepin-2-one
Step A - Preparation of 3-(Benzyloxycarbonyl)-amino-7-chloro-2,3-
dihydro-I -methyl-5-phenyl-1 H-1,4-benzodiazepin-2-one
A solution of 3-(benzyloxycarbonyl)-amino-7-chloro-2,3-dihydro-5-
phenyl-1H-1,4-Benzodiazepin-2-one (1 eq; Neosystem) in DMF was cooled to
0°C and treated with potassium tert-butoxide { 1 eq; 1.0M solution in
THF). The
resultant yellow solution was stirred at 0°C for 30 minutes then
quenched with
methyl iodide ( 1.3 eq). After stirring an addition 25 minutes the reaction
was
diluted with methylene chloride and washed with water and brine. The organic
phase was dried over Na,S04, filtered, and concentrated. The residue was
purified via HPLC chromatography eluting with a gradient of 20-X30% ethyl
acetate/hexanes.
C,4H,oC1N3O3 (MW = 433.92); mass spectroscopy 433.
. _.".~.. ,
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Anal. calcd for CZQHZoC1N3O3: C, 66.44; H, 4.65; N, 9.68. Found: C,
66.16; H, 4.50; N, 9.46.
Step B - Preaaration of 3-Amino-7-chloro-1 3-dih~dro-1-methyl-5-
phenyl-2H-1,4-benzodiazepin-2-one
Following General Procedure 8-B using 3-(benzyloxycarbonyl)-amino-7-
chloro-2,3-dihydro-1-methyl-5-phenyl-1 H-1,4-benzodiazepin-2-one, the title
intermediate was prepared as a white foam which was used immediately in Step
C.
Step C - Preparation of 3-fN'-tert-Butylcarbamate)-L-alani_ny_I1-
amino-7-chloro-1,3-dihydro-1-methyl-5-phenyl-2H-1 4-
benzodiazepin-2-one
Following General Procedure III-D using N-Boc-L-alanine and 3-amino-
1 S 7-chloro-1,3-dihydro-1-methyl-5-phenyl-21-1-1,4-benzodiazepin-2-one, the
title
intermediate was prepared as a white foam.
C,4HZ8C1N4O4 (MW = 471.18); mass spectroscopy 47I
Anal. calcd for C24H~gC1N404: C, 61.21; H, 5.78; N, 11.90. Found: C,
61.24; H, 5.59; N, 11.67.
Step D - Preparation of 3-(L-Alaninyl)amino-7-chloro-1 3-dihydro-
1-methyl-5-phenyl-2H-1 4-benzodiazepin-2-one
Following General Procedure 8-C using 3-[N'-tert-butylcarbamate)-L-
alaninyl]-amino-7-chloro-1,3-dihydro-1-methyl-5-phenyl-2H-1,4-benzodiazepin-
2-one, the title intermediate was prepared as a white foam. The crude material
was used immediately.
Example 8-D
Synthesis of
. 30 3-(L-Alaninyl)amino-7-bromo-2,3-dihydro-
1-methyl-5-(2-flnorophenyl)-1H-1,4-benzodiazepin-2-one
Step A - Preparation of 3-(Benzyloxycarbonyl)-amino-7-bromo-2,3-
dihydro-1-methyl-5-(2-fluorophenyl)-1 H-1,4-
benzodiazepin-2-one
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Following General Procedure 8-A using 3-(benzyloxycarbonyl)-amino-7-
bromo-2,3-dihydro-5-(2-fluorophenyl)-1 H-1,4-benzodiazepin-2-one (Neosystem},
the title intermediate was prepared as a white foam.
Cz4H,9BrFN3O3 (MW = 496.36); mass spectroscopy 497:
Anal. calcd for C~qH,9BrFN303: C, 58.08; H, 3.86; N, 8.47. Found: C,
57.90; H, 4.15; N, 8.20.
Step B - Preparation of 3-Amino-7-bromo-1,3-dihvdro-I-methyl-5-
(2-fluorophenyl)-2H-1,4-benzodiazepin-2-one
Following General Procedure 8-B using 3-(benzyloxycarbonyl)-amino-7-
bromo-2,3-dihydro-1-methyl-5-(2-fluorophenyl)- I H-1,4-benzodiazepin-2-one,
the
title intermediate was prepared as a white foam which was used immediately in
Step C.
Step C - Preparation of 3-[N'-(tert-Butylcarbamate)-L-alaninyll-
amino-7-bromo-1 (3-dihydro-1-methyl-5-(2-fluorophenyl)-
2H-1,4-benzodiazepin-2-one
Following General Procedure III-D using N-Boc-L-alanine (Novo) and 3-
amino-7-bromo-1, 3-dihydro-1-methyl-5-( 2-fluorophenyl)-2H-1,4-benzodiazepin-
2-one, the title intermediate was prepared as a white foam.
CzaH26BrFN4O4 (MW = 533.12); mass spectroscopy 533.2.
Anal. calcd for C~4H,6BrFN404: C, 54.04; H, 4.91; N, 10.50. Found: C,
53.75; H, 4.92; N, 10.41.
Step D - Preparation of 3-(L-Alanin~)-amino-7-bromo-1,3-dihydro-
1-methyl-5-(2-fluorophenvl)-2H-1,4-benzodiazepin-2-one
Following General Procedure 8-C using 3-[N'-(tent-butylcarbamate)-L-
alaninyl]-amino-7-bromo-1,3-dihydro-1-methyl-5-(2-fluorophenyl)-2H-1,4-
benzodiazepin-2-one, the title intermediate was prepared as a white foam. The
crude material was used immediately.
Example 8-E
Synthesis of
,. ,
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3-(N'-Methyl-L-alaninyl)-amino-2,3-dihydro- _
1-methyl-5-phenyl-IH-1,4-benzodiazepin-2-one
Step_A - Preparation of 3-fN'-(tert-Butylcarbamate)-N'-methyl-L-
alaninyll-amino-2,3-dihydro-1-meth~phenyl-1 H-1 4-
benzodiazenin-2-one _
Following General Procedure III-D and using (S)-3-amino-1,3-dihydro-I-
methyl-5-phenyl-2H-1,4-benzodiazepin-2-one (Example 8-B) and N-tert-Boc-N-
methyl-alanine (Sigma), the title intermediate was obtained as a white solid.
C,SH3oN404 (MW = 450.2); mass spectroscopy (M+I) 451.2.
Anal. calcd for C25H3oN40,: C, 66.65; H, 6.71; N, 12.44. Found: C,
66.66; H, 6.89; N, 12.21.
Step A - Preparation of 3-(N'-Methyl-L-alaninyl)-amino-~ 3-
dihydro-1-methyl-5-phenyl-1 H-1 4-benzodiazepin-2-one
Following General Procedure 8-C and using 3-(N'-(tert-butylcarbamate)-
N'-methyl-L-alaninyl)-amino-2,3-dihydro-1-methyl-5-phenyl-1 H-1,4-
benzodiazepin-2-one, the title intermediate was prepared as a white foam.
C,oH,2N402 (MW =350.46); mass spectroscopy (M+1) 351.4.
Anal. calcd for C~pfl"N40,: C, 68.55; H, 6.33; N, 15.99. Found, C,
68.36; H, 6.20; N, 15.79.
Example 8-F
Synthesis of
3-(L-Alaniny!)amino-7-chloro-2,3-dihydro
I-methyl-S-(2-chlorophenyl)-1H-1,4-benzodiazepin-2-one
Step A - Preparation of 3-(Benzyloxycarbonyl)-amino-7-chloro-2 3-
dihvdro-1-meths 2-chloro-phenyl)-1 H-1 4-
benzodiaz~in-2-one
Following General Procedure 8-A using 3-(benzyloxycarbonyl)-amino-7-
chlaro-2,3-dihydro-5-(2-chlorophenyl)-1H-1,4-benzodiazepin-2-one (Neosystem),
the title intermediate was prepared as a white solid having a melting point of
232-233°C.
Cz4H,gC12N30; (MW = 468.36); mass spectroscopy 468.
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~H NMR (300 MHz, CDCl3): S = 7.67 ( 1 H, m), 7.52-( 1 H; dd,
J=2.4, 8.7 Hz), 7.42-7.26 (9H, m), 7.07 ( 1 H, d, J=2.4 Hz), 6.70 ( I H, d,
J=8.3
Hz), 5.35 (1H, d, J=8.4 Hz), 5.14 (2H, ABq, J=19.6 Hz), 3.47 (3H, s).
~3C NMR (75 MHz, CDC13): 8 = 166.66, 165.65, 155.72, 140.52, 136.99,
136.0, 132.87, 131.99, 131.47, 131.40, 131.38, 131.16, 130.54, 130.06, 128.45,
128.08, 128.03, 127.72, 127.22, 123.28, 122.01, 68.95, 67.02, 35.32.
Step B - Preparation of 3-Amino-7-chloro-1.3-dihydro-1-methyl-5-
(2-chlorophenyl)-2H-I ,4-benzodiazepin-2-one
Following General Procedure 8-B using 3-(benzyloxycarbonyl)-amino-7-
chloro-2,3-dihydro-1-methyl-5-(2-chlorophenyl )- I H-1,4-benzodiazepin-2-one,
the
title intermediate was prepared as a white foam which was used immediately in
Step C.
Step C - Preparation of 3-[N'-(tert-Butylcarbamate)-L-alaninyll-
amino-7-chloro-1,3-dihydro- i -methyl-5-(2-chlorophenyl)-
2H-1,4-benzodiazepin-2-one
Following General Procedure III-D using N-Boc-L-alanine and 3-amino-
7-chloro-1,3-dihydro-1-methyl-5-(2-chlorophenyl)-2H-1,4-benzodiazepin-2-one,
the title intermediate was prepared as a white foam.
Cz4H~6C1~N4O4 (MW = 505.44); mass spectroscopy 505.2.
Step D - Preparation of 3-(L-Alaninyl)-amino-7-chloro-1,3-dihydro-
1-methyl-5-(2-chlorophenyl)-2H-1,4-benzodiazepin-2-one
Following General Procedure 8-C using 3-[N'-(tert-butylcarbamate)-L-
alaninyl]-amino-7-chloro-1,3-dihydro-1-methyl-5-(2-chlorophenyl)-2H-1,4-
benzodiazepin-2-one, the title intermediate was prepared as a white foam. The
crude material was used immediately.
Example 8-G
Synthesis of
3-(L-Alaninyl)amino-5-cyclohexyl-2,3-dihydro
1-methyl-1H-1,4-Benzodiazepin-2-one
.r... . .. r
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Step A - Preparation of 3-(Benzyloxycarbonyl)-amino-5-
cylclohexyl-2,3-dihydro-1-methyl-1 H-1 4-benzodiazepin-2-
one
Following General Procedure 8-A using 3-(benzyloxycarbonyl)-amino-5-
cyclohexyl-2,3-dihydro-1H-1,4-benzodiazepin-2-one (Neosystem), the title
intermediate was prepared as a white solid having a melting point of 205-
206°C.
CzqH~.,N3O3 (MW = 405.54); mass spectroscopy 405.
~H NMR (300 MHz, CDC13): 8 = 7.54 ( 1 H, d, J=7.9 Hz), 7.48 ( 1 H, d,
J=7.7 Hz), 7.36-7.26 (7H, m), 6.54 ( 1 H, d, J= 8.3 Hz), 5.15 ( 1 H, d, J=8.0
Hz),
5.09 (2H, ABq, J=17.1 Hz), 3.39 (3 H, s), 2.77 ( 1 H, m), 2.01 ( 1 H, bd,
J=13.6
Hz), 1.85 (1H, bd, J=12.4 Hz), 1.68-1.49 (4H, m), 1.34-1.02 (4H, m).
Step B - Preparation of 3-Amino-5-c clohexyl-1 3-dih d~o-1-
methyl-2H-1 4-benzodiazepin-2-one
Following General Procedure 8-B using 3-(benzyloxycarbonyl)-amino-5
cyclohexyl-2,3-dihydro-1-methyl-1 H-1,4-benzodiazepin-2-one, the title
intermediate was prepared as a white foam which was used immediately in Step
C.
C,6H~,N30 (MW+H = 272.1763); mass spectroscopy 272.1766
Step C - Preparation of 3-fN'-(tert-ButylcarbamateZ L-alaninvll-
amino-5-cyclohexyl-1 3-dihydro-1-methyl-2H-1 4-
benzodiazenin-2-one
Following General Procedure III-D. using N-Boc-L-alanine and 3-amino-
5-cyclohexyl-1,3-dihydro-1-methyl-2H-1,4-benzodiazepin-2-one, the title
intermediate was prepared as a white foam.
C~4H34N4Oq (MW = 442.62); mass spectroscopy (M+H) 443.2.
Step D - Preparation of 3-(L-Alaninyl)amino-5-cyclohex~l 3-
dihydro-1-methyl-2H-1 4-benzodiazepin-2-one
Following General Procedure 8-C using 3-[N'-(tert-butylcarbamate)-L-
alaninyl]-amino-5-cyclohexyl-1,3-dihydro-1-methyl-2H-1,4-benzodiazepin-2-one,
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the title intermediate was prepared as a white foam. The crude material was
used immediately.
C19~26N4~2 (M+H = 343.2136); mass spectroscopy found 343.2139.
Example 8-H
Synthesis of
3-(L-Alaninyl)amino-2,3-dihydro-1-methyl
7-nitro-5-phenyl-1H-1,4-benzodiazepin-2-one
Step A - Preparation of 2-[N-(a-Isopropylthio)-N'
(benzyloxycarbonyl)- lyg~ cinyl]-amino-5-nitrobenzophenone
A solution of a-(isopropylthio)-N-(benzyloxycarbonyl)glycine ( 1 eq;
prepared according to Zoller, V.; Ben-Ishai, D. Tetrahedron 1975, 31, 863.) in
dry THF was cooled to 0 °C and treated with oxalyl chloride ( I eq. )
and 3 drops
of DMF. After stirring for 15 minutes at 0°C, the cooling bath was
removed
and stirring continued at ambient temperature for 40 minutes. The solution was
recooled to 0°C. A solution of 2-amino-5-nitrobenzophenone (0.9 eq. ;
Acros)
and 4-methylmorpholine (2.0 eq.) in dry THF was added via cannulation to the
acid chloride. The cooling bath was removed and the reaction stirred at
ambient
for 5 hours. The reaction was diluted with methylene chloride and washed with
0.5 M citric acid, saturated aqueous NaHC03, and brine. The organic phase was
dried over Na2S04, filtered, and concentrated. The residue was purified via
preparative LC2000 eluting with a gradient of 15-X20% ethyl acetate/hexanes
giving an off white foam.
C26H25N3O6s (MW = 507.61 ); mass spectroscopy found 507.9.
Anal. calcd for C26H.,5N3O6S: C, 61.53; H, 4.96; N, 8.28. Found: C,
61.70; H, 4.99; N, 8.22.
Step B - Preparation of 2=jN-(a-Amino)-N'-(benzyloxycarbonyl)-
g_lycinyl3-amino-5-nitrobenzophenone
Ammonia gas was bubbled into a solution 2-[N-(a-isopropylthio)-N'-
(benzyloxycarbonyl)-glycinyl]-amino-5-nitrobenzophenone ( I eq) in THF at
0°C.
After 35 minutes mercury(II) chloride (l.l eq) was added. The ice bath was
removed and ammonia gas was continued to bubble through the suspension for 4
r
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hours. The bubbler was removed and the reaction continued to stir for I-6
hours.
. The mixture was filtered through celite washing with THF . The filtrate was
- concentrated-in vacuo. The crude solid was used in step C without further
purification.
- 5
Step C - Preparation of 3-(Benzyloxycarbonyl)-amino-2,3-dihvdro-
7-nitro-5-phenyl-1 H-1 4-benzodiazepin-2-one
2-[N-(a-Amino)-N'-(benzyloxycarbonyl)-glycinyl]-amino-5-
nitrobenzophenone ( 1 eq) was treated with glacial acetic acid and ammonium
acetate (4.7 eq). The suspension was stirred at ambient temperature for 21
hours. After concentrating the reaction in vacuo, the residue was partitioned
between ethyl acetate and 1 N NaOH. The aqueous layer was back-extracted
with ethyl acetate. The combined organics were washed with brine, dried over
Na,S04, filtered, and concentrated. The residue was purified via flash
chromatography eluting with a gradient of 2-~3% isopropyl alcohol/methylene
chloride.
C,3H,8N4O5 (MW = 430.45); mass spectroscopy found (M+H) 431.2.
Anal. calcd for C,3H,gN405: C, 64.18; H, 4.22; N, 13.02. Found: C,
64.39; H, 4.30; N, 13.07.
Step D - Preparation of 3-(Benz~ycarbonyl)-amino-2 3-dihy_dr_o_-
I -methyl-7-nitro-5-phenyl-1 H-1 4-benzodiazepin-2-one
Following General Procedure 8-A and using 3-(benzyloxycarbonyl)-
amino-2,3-dihydro-7-nitro-5-phenyl-1 H-1,4-benzodiazepin-2-one, the title
intermediate was prepared as a yellow foam.
Cz4H,oN405 (MW = 444.48); mass spectroscopy found (M+H) 445.2.
Anal. calcd for CZQHzoN4O5: C, 64.86; H, 4.54; N, 12.60. Found: C,
. 65.07; H, 4.55; N, 12.46.
Step E - Preparation of 3-Amino-1 3-dihvdro-1-methyl-7-nitro 5
phenyl-2H-1 4-benzodiazepin-2-one
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Following General Procedure 8-B and using 3-(benzyloxycarbonyl)-
amino-2,3-dihydro-1-methyl-7-nitro-5-phenyl-1H-1,4-benzodiazepin-2-one, the
title intermediate was prepared as a yellow foam which was used immediately in
Step F. -
Step F - Preparation of 3 jN'-(tert-Butylcarbamate)-L-alaninyl]-
amino-2, 3-dihydro-1-methyl-7-nitro-5-phenyl-1 H-1,4-
benzodiazepin-2-one
Following General Procedure III-D using N-Boc-L-alanine and 3-amino-
1,3-dihydro-1-methyl-7-nitro-5-phenyl-2H-1,4-benzodiazepin-2-one, the title
intermediate was prepared as a yellow solid.
C~4H,,NSO6 (MW = 481.56); mass spectroscopy found (M+H) 482.3.
Anal. calcd for C24H,,NSO6: C, 59.88; H, 5.61; N, 14.55. Found: C,
60.22; H, 5.75; N, 13.91.
Step G - Preparation of 3-(L-Alaninyl)-amino-2,3-dihydro-1-methyl
7-nitro-5-phenyl-1 H-1,4-benzodiazepin-2-one
Following General Procedure 8-C using 3-[N'-(tert-butylcarbamate)-L-
alaninyl]-amino-2, 3-dihydro-1-methyl-7-nitro-5-phenyl-1 H-1,4-benzodiazepin-2-
one, the title intermediate was prepared as a yellow foam. The crude material
was used immediately.
Example 8-I
Synthesis of
3-(L-Alaninyl)amino-2,3-dihydro-1-methyl-
5-(2-fluorophenyl)-1H-1,4-benzodiazepin-2-one
Step A - Preparation of 3-Amino-1,3-dihydro-1-methyl-S-(2-
fluorophenyl)-2H-1,4-benzodiazepin-2-one
A flask was charged with 3-(benzyloxycarbonyl)-amino-7-bromo-2,3-
dihydro-1-methyl-5-(2-fluorophenyl)-1H-1,4-benzodiazepin-2-one (1 eq.;
Example 8-D, Step A} and 10% palladium on carbon. Methanol was added, and
the flask was placed under a balloon of H,. The reaction was stirred for 21
.. ..._.. _.-._._._.......,.....,..w....~"..H...... ,_. T.
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hours. The mixture was filtered through celite- washing with methanol. The
filtrate was concentrated to a white solid.
- C,6H,4FN30 (MW = 283.33); mass spectroscopy found (M+H) 284.1.
Step B - Preparation of 3-fN'-(tert-Butylcarbamate)-L-alaninyll-
amino- I .3-dihydro- I -methyl-5-(2-fluorophenyl)-2H-I 4=
benzodiazepin-2-one
Following General Procedure III-D using N-Boc-L-alanine and 3-amino-
1,3-dihydro-1-methyl-5-(2-fluorophenyl)-2 H-1,4-benzodiazepin-2-one, the title
intermediate was prepared as a white solid.
C~4HZ.,FN404 (MW = 454.50); mass spectroscopy found (M+H) 455.4.
Anal. calcd for C~QH,~FN404: C, 63.44; H, 5.95; N, 12.33. Found: C,
63.64; H, 6.08; N, 12.16.
Step C - Preparation of 3-(L-Alaninyl)-amino-7-bromo-1 3-dih~o ,
I -methyl-5-(2-fluorophenyl)-2H-1 4-benzodiazepin-~-one
Following General Procedure 8-C using 3-[N'-(tert-butylcarbamate)-L-
alaninyl]-amino- I ,3-dihydro-1-methyl-5-(2-fluorophenyl)-2H-1,4-benzodiazepin-
2-one, the title intermediate was prepared as a white foam. The crude material
was used immediately.
Example 8-J
Synthesis of
3-(L-Alaninyl)-amino-2,3-dihydro
1-methyl-5-(3-fluorophenyt)-1H-1,4-benzodiazepin-2-one
Step A - Preparation of 2-Amino-3'-fluorobenzo~henone
A solution of 3-bromofluorobenzene (1 eq.) in THF was cooled to -
78°C
under nitrogen and treated with tert-butyllithium (2.05 eq., 1.6 M solution in
pentane) at a rate of 40 ml/h. The internal temperature did not rise above -
74°C. The orange solution was stirred at -78°C for 30 minutes
prior to the
addition of anthranilonitrile (0.6 eq.) as a solution in THF. The reaction was
warmed to 0°C and stirred for 2 hours. 3N HCl was added to the mixture
and
stirring continued for 30 minutes. The reaction was diluted with ethyl acetate
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and the layers were separated. The aqueous layer was back-extracted thrice
with
ethyl acetate. The combined extracts were washed with brine, dried over
Na,S04, filfered, and concentrated. The residue was purified via HPLC eluting
with 93:7 hexanes/ethyl acetate.
C,3H'QFNO (MW = 215.24); mass spectroscopy found (M+H) 216.3.
'H NMR (300 MHz, CDC13) d 7.44-7.19 (6H, m), 6.74 (1H, d, J=8.0
Hz), 6.61 ( 1 H, dd, J=0.94, 7.9 Hz), 6. I 0 (2H, bs).
Step B - Preparation of 2-[N-(a-Isopropylthio -
(benzyloxycarbonyl)-glycinyll-amino-3'-
fluorobenzophenone
A solution of a-(isopropylthio)-N-(benzyloxycarbonyl)glycine ( 1 eq;
prepared according to Zoller, V.; Ben-Ishai, D. Tetrahedron 1975, 31, 863.) in
dry THF was cooled to 0°C and treated with oxalyl chloride (1 eq.) and
3 drops
of DMF. After stirring for 15 minutes at 0°C, the cooling bath was
removed
and stirring continued at ambient temperature for 40 minutes. The solution was
recooled to 0°C. A solution of 2-amino-3'-fluorobenzophenone (0.9 eq.}
and 4-
methylmorpholine (2.0 eq.) in dry THF was added via cannulation to the acid
chloride. The cooling bath was removed and the reaction stirred at ambient for
5 hours. The reaction was diluted with methylene chloride and washed with 0.5
M citric acid, saturated aqueous NaHC03, and brine. The organic phase was
dried over Na2S04, filtered, and concentrated. The residue was purified via
preparative LC2000 eluting with a gradient of 15-X20% ethyl acetate/hexanes
giving an off white foam.
CZ6HzsNzOaS (MW = 480.60); mass spectroscopy found (M+NH4+) 498.3.
' H NMR (300 MHz, CDCl3) d 11.39 ( 1 H, s}, 8.59 ( 1 H, d, J=6.0 Hz),
7.63-7.55 (2H, m), 7.48-7.27 (9H, m), 7.14 ( 1 H, dt, J=1.2, 8.4 Hz), 5.94 ( 1
H, d,
J=7.2 Hz), 5.58 ( 1 H, d, J=8.7 Hz), 5.17 (2H, ABq, J=I4.7 Hz), 3.25 { I H,
sep,
J=6.6 Hz), 1.44 (3H, d, J=6.0 Hz), 1.28 (3H, d, J=6.6 Hz).
Step C - Preparation of 2-[N-(a-Amino)-N'~-(benzyloxvcarbonyl)-
~lycinyll-amino-3'-fluorobenzophenone
ru.. . ~ ,
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Ammonia gas was bubbled into a solution 2-[N-(a-isopropylthio)-N'-
(benzyloxycarbonyl}-glycinyl]-amino-3'-fluorobenzophenone ( 1 eq) in THF at
0°C. After 35 minutes mercury(II) chloride ( 1.1 eq) was added. The ice
bath
was removed and ammonia gas was continued to bubble through the suspension
- 5 for 4 hours. The bubbler was removed and the reaction continued to stir
for 16
hours. The mixture was filtered through celite washing with THF. The filtrate
was concentrated in vacuo. The crude solid was used in step D without further
purification.
Step D - Preparation of 3-(Benz~ycarbonyl)-amino-2 3-dihydro-
5-(3-fluorophen~)-1 H-1 4-benzodiazepin-2-one
2-[N-(a-Amino)-N'-(benzyloxycarbonyl)-glycinyl]-amino-3'-
fluorobenzophenone ( 1 eq) was treated with glacial acetic acid and ammonium
acetate (4.7 eq). The suspension was stirred at ambient temperature for 21
hours. After concentrating the reaction in vacuo, the residue was partitioned
between ethyl acetarte and 1 N NaOH. The aqueous layer was back-extracted
with ethyl acetate. The combined organics wire washed with brine, dried over
Na,S04, filtered, and concentrated. The residue was purified via flash
chromatography eluting with a gradient of 2-~3% isopropyl alcohol/methylene
chloride.
C,3H,8FN303 (MW = 403.44); mass spectroscopy found (M+H) 404.4.
Anal. calcd for C,3H,8FN303~O.SH~O; C, 66.98; H, 4.64; N, 10.18.
Found: C, 67.20; H, 4.64; N, 9.77.
Step E - Preparation of 3-(Benzyloxycarbonyl)-amino-2 3-dihydro-
1-methyl-5-(3-fluorophenyl)-1 H-1 4-benzodiazenin-2-one
Following General Procedure 8-A and using 3-(benzyloxycarbonyl)
, amino-2,3-dihydro-5-(3-fluorophenyl)-1H-1,4-benzodiazepin-2-one, the title
intermediate was prepared as a yellow foam.
C~alIzoFN~03 (MW = 417.47); mass spectroscopy found (M+H) 418.3.
. Anal. calcd for C,4H,oFN303: C, 69.06; H, 4.83; N, 10.07. Found: C,
69.33; H, 4.95; N, 9.82.
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Step F - Preparation of 3-Amino-1,3-dihydro-1-methyl-5-(3-
fluorophenyl)-2H-1,4-benzodiazepin-2-one
Following General Procedure 8-B and using 3-(benzyloxycarbonyl)-
amino-2,3-dihydro-1-methyl-5-(3-fluorophenyl)-1H-1,4-benzodiazepin-2-one, the
title intermediate was prepared as a yellow foam which was used immediately in
Step G.
Step G - Preparation of 3-(N'-(tert-But~carbamate)-L-alaninyll
amino-2.3-dihydro-1-methyl-5-(3-fluorophenyl)-1 H-1 (4
benzodiazepin-2-one
Following General Procedure III-D using N-Boc-L-alanine and 3-amino-
1,3-dihydro-1-methyl-5-(3-fluorophenyl}-2H-1,4-benzodiazepin-2-one, the title
intermediate was prepared as a yellow solid.
C,4HZ~FN404 (MW = 454.50); mass spectroscopy found (M+H) 455.3.
Anal. calcd for Cz4H~,FN404: C) 63.42; H, 5.99; N, 12.33. Found: C,
b3.34; H, 6.01; N, 12.08.
Step H - PreQaration of 3-(L-Alaninyl)-amino-2 3-dihydro-1-methyl-
5-(3-fluorophenyl)-I H-1 (4-benzodiazepin-2-one
Following General Procedure 8-C using 3-[N'-(tent-butylcarbamate)-L-
alaninyl]-amino-2,3-dihydro-1-methyl-5-(3-fluorophenyl)-1 H-1,4-benzodiazepin-
2-one, the title intermediate was prepared as a yellow foam. The crude
material
was used immediately .
Example 8-K
Synthesis of
3-(L-Alaninyl)amino-2,3-dihydro-I-methyl
5-(4-fluorophenyi)-1H-1,4-benzodiazepin-2-one
Step A - Preparation of 2-Amino-4'-fluorobenzophenone
A solution of 4-bromofluorobenzene (1 eq.) in THF was cooled to -
78°C
under nitrogen and treated with tert-butyllithium (2.05 eq., 1.6 M solution in
pentane) at a rate of 40 ml/h. The internal temperature did not rise above -
74°C. The orange solution was stirred at -78°C for 30 minutes
prior to the
_._...._~...__..,~..___
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addition of anthranilonitrile (0.6 eq.) as a solution in THF. The reaction was
warmed to 0°C and stirred for 2 hours. 3N HCl was added to the mixture
and
stirring continued for 30 minutes. The reaction was diluted with ethyl acetate
and the layers were separated. The aqueous layer was back-extracted thrice
with
- 5 ethyl acetate. The combined extracts were washed with brine, dried over
Na~S04, filtered, and concentrated. The residue was purified via HPLC eluting
with 93:7 hexanes/ethyl acetate.
C,;H,°FNO (MW = 215.24); mass spectroscopy found (M+H) 216.3.
Anal. calcd for C,3H,oFNO: C, 72.55; H, 4.68; N, 6.51. Found: C,
72.80; H, 4.51; N, 6.74.
Step B - Preparation of 2-IN-(a-Isopronvlthio)-N'-
(benzyloxycarbonyll-gl~yll-amino-4'-
fluorobenzophenone
A solution of a-(isopropylthio}-N-(benzyloxycarbonyl)glycine (1 eq;
prepared according to Zoller, V.; Ben-Ishai, D. Tetrahedron 1975, 31, 863.) in
dry THF was cooled to 0°C and treated with oxalyl chloride ( 1 eq.) and
3 drops
of DMF. After stirring for 15 minutes at 0°C, the cooling bath was
removed
and stirring continued at ambient temperature for 40 minutes. The solution was
recooled to 0°C. A solution of 2-amino-4'-fluorobenzophenone (0.9 eq.)
and 4-
methylmorpholine (2.0 eq.) in dry THF was added via cannulation to the acid
chloride. The cooling bath was removed and the reaction stirred at ambient for
5 hours. The reaction was diluted with methylene chloride and washed with 0.5
M citric acid, saturated aqueous NaHC03, and brine. The organic phase was
dried over Na2S04, filtered, and concentrated. The residue was purified via
preparative LC2000 eluting with a gradient of 15-X20% ethyl acetate/hexanes
giving an off white foam.
CZ6H~SN,O4S (MW = 480.60); mass spectroscopy found (M+NH4+) 498.2.
~ H NMR (3 00 Mflz, CDCl3) d I 1.28 ( 1 H, s), 8. 56 ( 1 H, d, J=8.4 Hz),
7.78-7.73 (2H, m), 7.61-7.53 (2H, m), 7.36-7.32 (5H, m), 7.20-7.14 (3H, m),
5.98 ( 1 H, d, J=7.5 Hz), 5.57 ( 1 H, d, J=7.8 Hz), 5.16 (2H, ABq, J=14.7 Hz),
3.25 (1H, sep, J=6.0 Hz), 1.43 (3H, d, J=6.3 Hz), 1.27 (3H, d, J=6.6 Hz).
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Step C - Preparation of 2-[N-(a-Amino)-N'-(benzvloxyearbonyl)-
gl r~cin~ll-amino-4'-fluorobenzophenone
Ammonia gas was bubbled into a solution 2-[N-(a-isopropylthio)-N'-
(benzyloxycarbonyl)-glycinyl]-amino-3'-fluorobenzophenone ( 1 _eq) in THF at
0°C. After 35 minutes mercury(II) chloride (1.1 eq) was added. The ice
bath
was removed and ammonia gas was continued to bubble through the suspension
for 4 hours. The bubbler was removed and the reaction continued to stir for 16
hours. The mixture was filtered through celite washing with THF. The filtrate
was concentrated in vacuo. The crude solid was used in step D without further
purification.
Step D - Preparation of 3-(Benzyloxycarbonyl)amino-2.3-dihydro-5
(4-fluoronhen~l-1 H-1 4-benzodiazepin-2-one
2-[N-(a-Amino}-N'-(benzyloxycarbonyl)-glycinyl]-amino-4'-
fluorobenzophenone ( 1 eq) was treated with glacial acetic acid and ammonium
acetate (4.7 eq). The suspension was stirred at ambient temperature for Li
hours. After concentrating the reaction in vacuo, the residue was partitioned
between ethyl acetate and 1 N NaOH. The aqueous layer was back-extracted
with ethyl acetate. The combined organics were washed with brine, dried over
Na,S04, filtered, and concentrated. The residue was purified via flash
chromatography eluting with a gradient of 2--~3% isopropyl alcohol/methylene
chloride.
C~3H,gFN303 (MW = 403.44); mass spectroscopy found (M+H) 404.4.
Anal. calcd for C,3H,gFN303~1.25H,0: C, 64.85; H, 4.85. Found: C,
64. 80; H, 4.5 5.
Step E - Preparation of 3-(Benzyloxycarbonyl)-amino-2.3-dihydro-
1-methyl-5-(4-fluorophenyl)-1 H-1 4-benzodiazenin-2-one
Following General Procedure 8-A and using 3-(benzyloxycarbonyl)
amino-2,3-dihydro-5-(4-fluorophenyl)-1H-1,4-benzodiazepin-2-one, the title
intermediate was prepared as a yellow foam.
C,4H2°FN3O3 (MW = 417.47); mass spectroscopy found (M+H) 418.2.
.. .....~. ,.
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Anal. calcd for C,4HzoFN303: C, 69.06; H, 4.83; N, 10.07. Found: C,
69.35; H, 4.93; N, 9.97.
Step F - Preparation of 3-Amino-1,3-dihydro-1-methyl-5-(4-
fluorophenyl)-2H-1.4-benzodiazepin-2-one
Following General Procedure 8-B and using 3-(benzyloxycarbonyl)-
amino-2,3-dihydro-1-methyl-5-(4-fluorophenyl)-1H-1,4-benzodiazepin-2-one, the
title intermediate was prepared as a yellow foam which was used immediately in
Step G.
Step G - Preparation of 3-(N'-(tert-Butylcarbamate)-L-alanin~l-
amino-2, 3-dihydro- I -methyl-5-(3-fluorophen,Xl)-1 H-1 4-
benzodiazepin-2-one
Following General Procedure III-D using N-Boc-L-alanine and 3-amino-
1,3-dihydro-I-methyl-5-(3-fluorophenyl)-2I-I-1,4-benzodiazepin-2-one, the
title
intermediate was prepared as a yellow solid.
C24HZ,FN404 (MW = 454.50); mass spectroscopy found (M+H) 455.4.
Anal. calcd for C24H27FN4~4~ 1.SH,0: C, 59.86; H, 6.28; N, 11.64.
Found: C, 60.04; H, 5.62; N, 11.27.
Step H - Preparation of 3-(L-Alaninvl)-amino-2.3-dihydro-1-methyl-
5-(4-fluorophenyl)-1 H-1,4-benzodiazepin-2-one
Following General Procedure 8-C using 3-[N'-(tent-butylcarbamate)-L-
alaninyl]-amino-2,3-dihydro-1-methyl-5-(4-fluorophenyl)-1 H-1,4-benzodiazepin-
2-one, the title intermediate was prepared as a yellow foam. The crude
material
was used immediately.
Example 8-L
Synthesis of
3-(N'-L-Alaninyl)amino-2,3-dihydro-I-isobutyl-
5-phenyl-1H-1,4-benzodiazepin-2-one
Step A: 1,3-Dihydro-5-phenyl-2H-1,4-benzodiazepin-2-one (prepared
according to the procedure of M. G. Bock et al., J. Org. Chem. 1987, 52, 3232-
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3239) was alkylated with isobutyl iodide using General Procedure 8-G to afford
1,3-dihydro-1-isobutyl-5-phenyl-2H-1,4-benzodiazepin-2-one.
Step B: Following General Procedures 8-D and 8-F and using the
product from Step A, 3-amino-1, 3-dihydro- I -isobutyl-5-phenyl-2H- I ,4-
benzodiazepin-2-one was prepared.
Step C: The product from Step B and N-Boc-L-alanine (Sigma) were
coupled using General Procedure III-D, followed by removal of the Boc group
using General Procedure 8-J, to afford 3-(N'-L-alaninyl)amino-1,3-dihydro- I -
isobutyl-5-phenyl-2H-1,4-benzodiazepin-2-one.
By substituting isopropyl iodide, n-propyl iodide, cyclopropylmethyl
iodide and ethyl iodide for isobutyl iodide in Step A above, the following
additional intermediates were prepared:
3-(N'-L-alaninyl)amino-1,3-dihydro-1-isopropyl-5-phenyl-2H-1,4-
benzodiazepin-2-one
3-(N'-L-alaninyl)amino-I ,3-dihydro-1-propyl-5-phenyl-2H-1,4-
benzodiazepin-2-one
3-(N'-L-alaninyl)amino-1,3-dihydro-1-cyclopropylmethyl-5-phenyl-2H-
1,4-benzodiazepin-2-one
3-(N'-L-alaninyl)amino- I ,3-dihydro- I -ethyl-5-phenyl-2H-1,4-
benzodiazepin-2-one.
Example 8-M
Synthesis of
3-(N'-L-Alaninyl)amino-1-methyl=5-phenyl
1,3,4,5-tetrahydro-2H-1,5-benzodiazepin-2-one
Step A: 1,3,4,5-Tetrahydro-5-phenyl-2H-1,5-benzodiazepin-2-one (CAS
No. 32900-17-7) was methylated using General Procedure 8-I to afford I-
methyl-5-phenyl-1,3,4,5-tetrahydro-2H-1,5-benzodiazepin-2-one.
."r.,........ 9.
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DEMANDES OU BREVETS VOI.UMINEUX
LA PRESENTS PARTIE DE CETTE DEMANDS OU CE BREVET
COMPREND PLUS D'UN TOME.
CECI EST LE TOME ~-DE _~
NOTE: Pour les tomes additionels, veuillez contacter le Bureau canadien -des
brevets -
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