GROWTH HORMONE SECRETAGOGUES

SECRETAGOGUES D'HORMONES DE CROISSANCE

English Abstract

This invention relates to novel compounds which are useful in the modulation
of endogenous growth hormone levels in a mammal. The invention further relates
to novel intermediates for use in the synthesis of said compounds, as well as
novel processes employed in these syntheses. Also included are methods of
treating a mammal which include the administration of said compounds.

French Abstract

L'invention concerne de nouveaux composés convenant à la modulation de niveaux d'hormones de croissance endogènes chez un mammifère. L'invention concerne aussi de nouveaux intermédiaires servant à la synthèse de ces composés ainsi que de nouveaux procédés mis en oeuvre lors de ces synthèses. Font aussi l'objet de cette invention des procédés de traitement d'un mammifère consistant à lui administrer ces composés.

Claims

826



We claim:


1. A compound of formula I

Image

wherein:
A is C1-C6alkyl, aryl, C1-C6alkylaryl,
C1-C6alkyl (O) C1-C6alkylaryl, C1-C6alkyl (S) C1-C6alkylaryl,
indolyl, indolinyl, thienyl, (C1-C6alkyl)thienyl,
benzothienyl, benzofuranyl, naphthanyl, cyclohexyl, (C1-
C6alkyl) indolyl, (C1-C6alkyl)benzothienyl,
(C1-C6alkyl)naphthanyl, (C1-C6alkyl)benzofuranyl, and
(C1-C6alkyl) cyclohexyl;
B is NH2, NHR2, C1-C6alkylNH2, C1-C6alkylNHR1,
C1-C6alkylarylNH2, C1-C6alkylarylNHR1,
C1-C6alkylcyclohexylNH2, C1-C6alkylcyclohexylNHR1,
R1-piperidin-3-yl (C1-C6alkyl),
R1-piperidin-2-yl (C1-C6alkyl), R1-piperidin-4-yl (C1-C6alkyl),
R1-quinolin-2-yl (C1-C6alkyl),
R1-(2,4-dihydroquinolin-2-yl (C1-C6alkyl),
R1-isoquinolin-2-yl (C1-C6alkyl), and
R1-(2,4-dihydroisoquinolin-2-yl (C1-C6alkyl);
R1 is hydrogen, C1-C6alkyl, C1-C6alkyl (OH), or
C1-C6alkylidenyl (OH)R2;
R2 is C1-C6alkyl, C1-C6alkenyl,



827



C1-C6alkyl(O)C1-C6 alkyl, C(O)O-C1-C6 alkyl, aryl, or
C1-C6alkylaryl;
X is C1-C6alkylidenyl, O, S, NH, or N (C1-C6alkyl);
V is selected from the group consisting of



Image


828



and

Image


W is S, O,, NH, or CH2;
Y is N or CH;
Z is N or CH;
Y' is N or CH;
Z' is N or CH;
R4 and R5 are independently hydrogen, C1-C6alkyl,
aryl, C1-C6alkylaryl, C(O)O(C1-C6alkyl),
C(O)N(C1-C6alkyl)2, or C1-C6alkylCOR7;
R7 is hydrogen, C1-C6alkyl, pyrrolidinyl,
piperidinyl, homoproline, or proline;
D is hydrogen, C1-C6alkyl, C1-C6alkyl (aryl) 2,
C1-C6alkyl(O)(CO)C1-C6alkyl, C1-C6alkyl(O)(CO)N(C1-C6alkyl)2,
C1-C6alkylaryl, C(O)R6, C1-C6alkyl(O)R6, C1-C6alkyl (OH) , C1-C6
alkylC(O)R6, C1-C6alkylR6, aryl, (C1-C6alkyl) NHSO2(C1-C6alkyl),
(C1-C6alkyl)NHSO2(aryl);
R6 is H, C1-C6alkyl, aryl, naphthyl,
C1-C6alkylaryl, acetyl, NH2, NH (C1-C6alkyl),
NH(C1-C6alkyl)O(C1-C6alkyl) , NH(C1-C6alkyl)O(aryl), NH(C1-
C6alkyl), S(C1-C6alkyl), NH(C1-C6alkylidenyl)OCH3, NH(C1-
C6alkyl)aryl, NH(C3-C6cycloalkyl), NH(C1-C6alkyl)C(O)(C1-
C6alkyl), NH(C1-C6alkyl)NH(C1-C6alkyl), NH(C1-C6alkyl)NH(C1-
C6alkylaryl), NHSO2(C1-C6alkylaryl), NH(C1-C6alkyl)C(O)O(C1-
C6alkyl), NH(aryl), N(C1-C6alkyl)2, N(C1-C6alkyl)(aryl),
N(C1-C6alkyl)(C1-C6alkylaryl), O(C1-C6alkyl), O(aryl),
O(C1-C6alkylaryl), piperidinyl, piperidinyl-(C1-C6alkyl),
piperidinyl-C(O)NH (C1-C6alkyl), piperidinyl-C(O)NH(C1-
C6alkylaryl), piperidinyl-C(O)N(C1-C6alkyl)2, piperidinyl-C(O)
N(C1-C6alkyl)(aryl), piperidinyl-C(O)O(C1-C6alkyl),
pyrrolidinyl, piperidinyl-(C1-C6alkyl), pyrrolidinyl,



829



C(O)NH(aryl)-, pyrrolidinyl C(O)NH(C1-C6alkyl),
pyrrolidinyl- C(O)O(C1-C6alkyl) , pyrrolidinyl (C1-C6alkyl)O
(C1-C6alkyl), pyrrolidinyl C(O)NH(C1-C6alkyl)2-, pyrrolidinyl
C(O)NH (C1-C6alkylaryl)-, pyrrolidinyl C(O)NH (C1-C6alkyl)
aryyl-pyrrolinyl, morpholino, hexamethyleneimino,
heptamethyleneimino, quinolinyl, 2,4-dihydroquinolinyl,
1,2,3,4-tetrahydroquinolinyl, 2,4-dihydroisoquinolinyl,
1,2,3,4-tetrahydroisoquinolinyl, indolinyl, an amino acid
selected from the group consisting of proline, homoproline,
glycine, alanine, valine, leucine, isoleucine, tyrosine,
tryptophan, phenylalanine, serine, threonine, asparagine,
glutamic acid, aspartic acid, lysine, arginine, glutamine,
histidine, cysteine, and methionine, or a nitrogen-
containing heterocycle selected from the group consisting of




830



Image


831



E is hydrogen, C1-C6alkyl, C(O)C1-C6alkyl, aryl,
(aryl)C(O)NR6, (aryl) (C1-C6alkyl)C(O)R6,
C1-C6alkyl aryl, C(O)aryl, C1-C6 alkylC(O)aryl, naphthyl,
C1-C6alkylnaphthyl, C(O)naphthyl, C1-C6alkylC(O)naphthyl,
heteroaryl, C1-C6alkylheteroaryl, C(O)heteroaryl, C1-C6
alkylC(O)heteroaryl, indanyl, C1-C6alkylindanyl,
C(O)indanyl, C1-C6alkylC(O)indanyl, cycloalkyl;
or D and E combine to form indanyl, fluorenyl, or
cycloalkyl, unsubstituted or substituted by hydroxyl, O(CO)
C1-C6alkyl, C(O)R6 or O(CO)R6;
G is hydrogen, C1-C6alkyl, aryl, C1-C6alkylaryl,
and C1-C6alkenyl;
J is hydrogen, C1-C6alkyl, aryl, and
C1-C6alkylaryl;
L is hydrogen, C1-C6alkyl, C(O)OC1-C6alkyl, aryl,
C1-C6alkylaryl, C(O)OC1-C6alkylaryl, C1-C6alkenyl, -F, and
-CN, C1-C6alkyl-OH, C1-C6alkyl-O-C1-C6alkyl,
C1-C6alkyl-C(O)R6;
or a pharmaceutically acceptable salt or solvate thereof.
2. A compound according to Claim 1 wherein A is
selected from the group consisting of


Image



832



3. A compound according to Claim 1 wherein B is

Image

4. A compound according to Claim 1 wherein X is
NH.
5. A compound according to Claim 1 wherein V is
selected from the group consisting of

Image

6. A compound according to Claim 1 wherein D is
-C(O)R6, and R6 is
1-pyrrolidinyl, 1-piperidinyl, 4-methyl-1-piperidinyl,

Image

7. A compound according to Claim 1 wherein E is




833


Image

8. A compound according to Claim 1 wherein G is H.
9. A compound according to Claim 1 wherein J is H.
10. A compound according to Claim 1 wherein L is H or
CH3.
11. A method for increasing the level of endogenous
growth hormone in a human or an animal which comprises
administering to said human or animal an effective amount of
a compound of Claim :1.
12. A method according to Claim 16 which further
comprises administering to a patient a bone antiresorptive
agent.
13. A method according to Claim 29 wherein said bone
antiresorptive agent is a bisphosphonate.
14. A method for the treatment or prevention of a
physiological condition which may be modulated by an
increase in endogenous growth hormone which comprises



834



administering to an animal in need of said treatment an
effective amount of a compound of Claim 1.
15. A pharmaceutical formulation which comprises as an
active ingredient a compound of Claim 1, or a
pharmaceutically acceptable salt or solvate thereof,
associated with one or more pharmaceutically acceptable
carriers, diluents, or excipients.

Description

CA 02340344 2001-02-12
s
F
DEMANDES OU BREVETS VOLUMINEUX
LA PRESENTF PARTIE DE C>=TT>: DEMANDE OU CE BREVET
COMPREND PLUS D'UN TOME.
CECI EST LE TOME . ~ DE
NOTE: Pour les tomes additionels, veuiilez cantacter le Bureau canadien des
brevets
JUMBO APPLICATIONS/PAT>=NTS
TlilS SECT10N OF THE AP?LICAT10N/PATENT CONTAINS MORE
THIS IS VOLUME ~_ OF J
I110TE:'For additionay voiumes-ple.ase cnntact'the Canadian Patent Office


CA 02340344 2001-02-12
WO 00/10565 PCT/US99/03525
- 1 -
Title
GRO~TTH HORMONE SECRETAGOGUES
Growth hormone is a secretory protein of the pituitary
gland of animals having wide ranging developmental effects
on the organism. Artificial manipulation of growth hormone
levels has been demonstrated to have significant therapeutic
utility. Human growth hormone supplementation has been
shown to be an effecaive treatment for growth hormone
deficiencies and their related disease states in humans.
Apart fram this app7.ication, studies have uncovered new and
significant properties of growth hormone which lend further
importance to the ability to control growth hormone levels.
For example, recent clinical studies indicate that growth
hormone supplementation may be useful in combating the
maladies of aging in. humans. Elevated growth hormone levels
in animals have been. shown to result in increased lean
muscle mass. One application of this latter observation
could result in higher production of leaner meat products or
in the production of larger and/or stronger animals.
While growth hormone is naturally produced by the
pituitary gland, the secretion of growth hormone into the
bloodstream is controlled by a second protein, Growth
Hormone Releasing Factor (GRF). This hormone is also
commonly known in the art as somatocrinin, Growth Hormone
Releasing Hormone (GHRH), and Growth Releasing Hormone
( GRH ) .
There are two ways to approach the problem of
increasing circulating levels of growth hormone: (1)
increase the level of human growth hormone in the organism
directly or (2) increase the organism's natural tendency to
produce growth hormone. The latter strategy may be achieved
via supplementation with GRF. GRF has been demonstrated to
increase the circulatory levels of growth hormone in vivo.
(Rivier, et al., Nature (London). 300:276 (1982). The
SUBSTITUTE SHEET (RULE 26)


CA 02340344 2001-02-12
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effect of GRF, inclvuding structural analogs thereof, on
growth hormone production has been widely studied. A
primary obstacle to the use of GRF as a direct supplement is
its short lifespan _in vivo. L.A. Frohman, et al. , Journal
of Clinical Investigation, 78:906 (1986) . More potent
and/or longer lasting GRF molecules are therefore desirable
for the development of effective human therapeutic or animal
husbandry agents.
The structure of GRF has been modified in numerous ways
resulting in longer lasting and/or more potent GRF analogs.
It has been demonstrated that the first 29 amino acids from
the N-terminus are sufficient to retain full GRF activity.
Speiss, et al., Biochemistry, 21:6037 (1982). One strategy
has been the incorporation of novel D-amino acid residues in
various regions of the GRF molecule. V.A. Lance, et al.,
Biochemical and Biophysical Research Communications, 119:265
(1984) ; D.H. Coy, et al., Peptides, 8 (suppl. 1) : 49 (1986) .
Another strategy has modified the peptide backbone of GRF by
the incorporation of peptide bond isosteres in the N-
terminal region. D. 'rourwe, Janssen. Chim. Acta, 3:3
(1985); S.J. Hocart, et al., Journal of Medicinal Chemistry,
33:1954-58 (1990). A series of very active analogs of GHRH
is described in European Patent Publication 511,003,
published October 28, 1992.
In addition to t:he actions of GHRH there are various
ways known to releases growth hormone. For example,
chemicals such as arg~inine, z-3,4-dihydroxyphenylalanine (L-
DOPA), glucagon, vasopressin, and insulin-induced
hypoglycemia, as well as activities such as sleep and
exercise, indirectly cause growth hormone to be released
from the pituitary by acting in some fashion on the
hypothalamus, perhaps either to decrease somatostatin
secretion or to increase the secretion of GHRH.
In cases where increased levels of growth hormone are
desired, the problem leas generally been solved by providing
exogenous growth hormone or by administering GHRH, or a
SUBSTITUTE SHEET (RULE 26)


CA 02340344 2001-02-12
WO 00/10565 PCT/US99/03525
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related peptidyl compounds which stimulates growth hormone
production or release. In either instance the peptidyl
nature of the compound has necessitated that it be
administered by injection.
Other compound:> have been developed which stimulate the
release of endogenous growth hormone, such as analogous
peptidyl compounds related to GHRH. These peptides, while
considerably smaller than growth hormones are still
susceptible to metabalic instability.
Administration of the hexapeptide growth hormone
releasing peptide-6 (GHRP-6) results in the secretion of
growth hormone in many species, including humans.
This peptide is one of a series of synthetic peptides, the
structures of which were based on the pentapeptide Met-
enkephalin. It has been shown that GHRP binds specifically
to the pituitary, although the binding does not involve the
opioid, GHRH, or the somatostatin receptors.
In recent years significant efforts have been taken to
develop nonpeptidyl analogs of this series of compounds.
Such compounds, termed growth hormone secretagogues, should
be orally bioavailable, induce the production or release of
growth hormone, and act in concert, or synergistically with
GHRH.
Representative growth hormone secretagogues are
disclosed in United States Patent 3,239,345; United States
Patent 4,036,979; United States Patent 4,411,890; United
States Patent 5,206,:?35; United States Patent 5,248,841;
United States Patent 5,310,737; United States Patent
5,310,017; European 1latent Publication 144,230; European
Patent Publication 513,974; Patent Cooperation Treaty Patent
Publication WO 94/07486; Patent Cooperation Treaty Patent
Publication WO 94/08583; Patent Cooperation Treaty Patent
Publication WO 94/13696; United States Serial Number
08/704,494, filed August 20, 1996, United States Serial
Number 08/700, 206, f5_led August 20, 1996, and Science,
260:1640-1643 (1993).
SUBSTITUTE SHEET (RULE 26j


CA 02340344 2001-02-12
WO 00/10565 PCT/US99/03525
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United States Patent 5,206,235, issued April 27, 1993,
describes a series of benzolactam compounds typified by the
following structure.
H
N NH 2
N ~ 0 H3C \CH 3
.N
\\
N
i
N
H
These compounds have demonstrated clinical activity in
humans in raising the growth hormone secretory levels. B.J.
Gertz, Journal of Clinical Endocrinology and Metabolism,
77:1393-1397 (1993) .
Another group of growth hormone secretagogues is
described in Patent Cooperation Treaty Patent Publication WO
94/13696, published .June 23, 1994. These compounds are
typified by the following two structures.
CH3 CH3
~ H
N~~ ~3+ CH3S03
0 0
N
I ~ ~ N~
S02CH3
SUBSTITUTE SHEET (RULE 26)


CA 02340344 2001-02-12
WO 00/10565 PC'T/US99/035Z5
- 5 _
H H
N ~N
O O
DT N N
H
The present invention provides a series of compounds
that have activity as growth hormone secretagogues. These
compounds are non-peptidyl in nature and are, therefore,
more metabolically stable than growth hormone, growth
hormone releasing hormone, or analogs of either of these
proteins. The compounds employed in the present invention
are preferred for hwnan pharmaceutical uses as well as
veterinary uses, particularly in cattle, swine, sheep,
poultry and fish.
The present invention relates to compounds of formula I
A N B
1D V
L- I
E
I
wherein:
A is C1-C6a:Lkyl, aryl, C1-Csalkylaryl,
C1-Csalkyl (O) C1-Csalkylaryl, C1-C6alkyl (S) C1-C6alkylaryl,
indolyl, indolinyl, t:hienyl, (C1-C6alkyl ) thienyl,
.SUBSTITUTE SHEET (RULE 28)


CA 02340344 2001-02-12
WO 00/10565 PCT/US99/03525
- 6 -
benzothienyl, benzofuranyl, naphthanyl, cyclohexyl, (C,-
C6alkyl) indoiyl, (C,-Csalkyl)benzothienyl,
(C,-C6alkyl)naphthanyl, (C,-C6alkyl)benzofuranyl, and
(C1-C6alkyl) cyclohe~;yl;
B is NHS, NHRl, C~-C6alkylNH2, C,-CsalkylNHRl,
C1-CsalkylarylNH2, C1-C6alkylarylNHRl,
C1-C6a1ky1cyclohexylNH2, C1-C6a1ky1cyclohexylNHRi,
Rl-piperidin-3-yl (C,-C6alkyl) ,
Rl-piperidin-2-yl (C,-C6alkyl) , Rl-piperidin-4-yl (C,-C6alkyl) ,
Rl-quinolin-2-yl (C1-Csalkyl) ,
R1- (2, 4-dihydroquinolin-2-yl (C~-C6alkyl) ,
Rl-isoquinolin-2-yl (C~-C6alkyl) , and
Rl- (2, 4-dihydroisoquinolin-2-yl (C,-Csalkyl) ;
Rl is hydrogen, C,-C6alkyl, C1-C6alkyl (OH) , or
C1-Csalkylidenyl (OH):R2;
RZ is C,-G,;alkyl, C1-Csalkenyl,
C1-C6alkyl (0) C1-C6 alkyl, C (O) O-C1-C6 alkyl, aryl, or
C1-C6a1 kylaryl ;
X is C1-C6alkylidenyl, 0, S, NH, or N(C1-C6alkyl) ;
V is selecaed from the group consisting of
SUBSTITUTE SHEET (RULE 28)


CA 02340344 2001-02-12
WO 00/10565 PGT/US99/03525
/\ /~ ~~ ~~ /~N
N~
I I ~ I
N ~N // N W W
/ Ra \ ~ 1
N~ Nw ~N /
RS N N~ N Y / Z
I i I i
J
\ / ~~ \ / ~ \ /
( I
H
-N~
H2 I
N ~ \
R4 N. N / NJ
I I
N NH Z , ~ /
I l
N
SUBSTITUTE SHEET (RULE 28)


CA 02340344 2001-02-12
WO 00/10565 PGT/US99/035Z5
and
(\
W is S, 0, NH, or CH2;
Y is N or CH;
Z is N or CH;
Y' is N or CH;
Z' is N or CH;
Rq and R5 are independently hydrogen, C,-C6alkyl,
aryl, C1-C6alkylaryl, C (0) 0 (C1-C6alkyl) ,
C (O) N (C1-C6alkyl ) 2, or C,-C6a1ky1COR,;
R, is hydrogen, Cl-C6alkyl, pyrrolidinyl,
piperidinyl, homopro:Line, or proline;
D is hydrogen, C1-C6alkyl,
C1-C6alkyl (O) (CO) C,-C6alkyl, C1-C6alkyl (O) (CO) N (C,-C6alkyl) 2,
C1-Csalkyl aryl, C (0) Rr" C,-C6alkyi (O) R6, C1-Csalkyl (OH) , C1-C6
alkylC (O) R6, C,-C6a1ky1R6, aryl, (C,-C6alkyl) NHSOZ (C1-Csalkyl) ,
( C1-C6alkyl ) NHSOZ ( aryl. ) ;
R6 is H, C1--Csalkyl, aryl, naphthyl,
C,-C6alkylaryl, acetyl, NH2, NH (C,-Csalkyl ) ,
NH (C,-C6alkyl) 0 (C1-Csalkyl) , NH (C1-C6alkyl) S (C1-C6alkyl) ,
NH (C,-C6alkylidenyl) OC:H3, NH (C,-C6alkyl) aryl,
NH (C3-C6 cycloalkyl) , NH (C1-C6alkyl) C (0) (C1-C6alkyl) ,
NH (C1-C6alkyl ) NH (C1-C6alkyl ) , NH (C1-C6alkyl ) NH (C1-Csalkyl aryl ) ,
NHSOZ (C1-C6alky1aryl) , NH (Ci-C6alkyl) C (0) 0 (C1-Csalkyl) ,
NH (naphthyl ) , N (Cl-C6al.kyl ) 2, N (C1-Csalkyl } ( aryl ) ,
N (C1-C6alkyl ) (C,-C6alkyl aryl ) , 0 ( CI-C6alkyl ) , O ( aryl ) ,
O (C,-C6alkylaryl) , piperidinyl,
piperidinyl-C (0} NH (C,--C6alkyl ) , piperidinyl-C (O) NH (C1-
C6alkylaryl) , piperid:inyl-C (0) N (C1-C6alkyl) 2,
piperidinyl-C (O) N (C1-C:falkyl) (aryl) ,
pyrrolidinyl, pyrrolidinyl C(0)NH(aryl)-,
SUBSTITUTE SHEET (RULE 28)


CA 02340344 2001-02-12
wo oonos6s Pcrius~ross2s
- g _
pyrrolidinyl C (0) NH (C1-Csalkyl) -,
pyrrolidinyl C (0) NH (C1-C6alkyl) 2-,
pyrrolidinyl C (O) NH (C,-C6alkylaryl) -,
pyrrolidinyl C (0) NH (C1-C6alkyl ) aryl-,
pyrrolinyl, morpholino, hexamethyleneimino,
heptamethyleneimino, quinolinyl, 2,4-dihydroquinolinyl,
1, 2, 3, 4-tetrahydroquinolinyl,
2,4-dihydroisoquino.linyl, 1,2,3,4-tetrahydroisoquinolinyl,
indolinyl, an amino acid selected from the group consisting
of proline, homopro:line, glycine, alanine, valine, leucine,
isoleucine, tyrosine, tryptophan, phenylalanine, serine,
threonine, asparagine, glutamic acid, aspartic acid,
lysine, arginine, glutamine, histidine, cysteine, and
methionine, or a nitrogen-containing heterocycle selected
from the group consisting of
SUBSTITUTE SHEET (RULE 28)


CA 02340344 2001-02-12
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- 10 -
N N N N N
H NCH3
N N N
NCl--C6alkyl NC1-C6alkylaryl
N N ,N N N N
C
S O ~N ~ 0 Naryl
S02CH3
N N N
.n
NL~ L'J
aryl
CN\aryl C (O) C1-C;~alkyl
N ~ aryl
N ,N N N
alkylaryl Oaryl aryl
aryl CN
E is hydrogen, C,-Csalkyl, C (0) C1-C6alkyl, aryl,
(aryl) C (0) NR6, (aryl) (C,-C6alkyl) C (0) R6,
C1-C6alkylaryl, C (0) aryl, C1-C6 alkylC (0) aryl, naphthyl,
C1-C6alkylnaphthyl, C(O)naphthyl,
C1-C6a1ky1C (0) naphthy7_, heteroaryl, C1-C6alkylheteroaryl,
C (O) heteroaryl, C1-C6 alkylC (0) heteroaryl, indanyl,
SUBSTITUTE SHEET (RULE 26~


CA 02340344 2001-02-12
wo oonos6s Pcrivs~ro35zs
- 11 -
C,-C6alkylindanyl, C (O) indanyl, C,-C6a1ky1C (O) indanyl,
cycloalkyl~
or D and E combine to form indanyl, fluorenyl, or
cycloalkyl;
G is hydrogen, C,-C6alkyl, aryl, C,-C:.alkylaryl,
and C,-C6alkenyl;
J is hydrogen, C,-Csalkyl, aryl, and
C,-Csalkylaryl;
L is hydrogen, C,-C6alkyl, C (O) OC,-C:alkyl, aryl,
C,-C6alkylaryl, C (O) OC,-Csalkylaryl, C,-C6alkenyl, -F, and
-CN, C,-C6alkyl-OH, C,-C6alkyl-O-C,-C6alkyl,
C,-C6alkyl-C (O) R6;
or a pharmaceutical:Ly acceptable salt or solvate thereof.
The present invention relates to compounds of formula I
H
I
A N H
X V
D' _V
~E
I
wherein:
A is C,-C6alkylaryl, C,-C6alkyl (O) C,-Csalkylaryl,
(C,-C6alkyl) indol-3y1, (C,-Csalkyl)benzothien-3y1, (C,-
Csalkyl)naphthan-2y1, (C,-Csalkyl)benzofuran- 3y1, and (C,-
Csalkyl) cyclohexyl;
B is C,-C6a1ky1NHR,, C,-C6alkylarylNHR,,
C,-Csalkylcyclohexyll~laR,, R,-piperidin-3-yl (C,-C6alkyl) ,
R,-piperidin-2-yl (C,-Cbalkyl) , R,-piperidin-4-yl (C,-C6alkyl) ,
R,-quinolin-2-yl (C,-Cf;alkyl) , R,- (2, 4-dihydroquinolin-2-yl (C,-
C6alkyl), R,-isoquino_Lin-2-yl (C,-Csalkyl) , and
R,- (2, 4-dihydroisoqui:nolin-2-yl (C,-C6alkyl) ;
SUBSTITUTE SHEET (RULE 26)


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Rl is hydrogen, C,-C6alkyl, C,-C6alkyl (OH) , or
C1-Csalkylidenyl (OH) R~;
R2 is C1-Coalkyl, C,-C6alkenyl, C,-C6alkyl (O) C,-C6
alkyl, C (0) 0-C1-C6 alkyl, aryl, or C1-C6alkylaryl;
X is C,-Cbalkylidenyl, O, S, NH, or N(C1-C6alkyl) ;
V is a nii=rogen-containing heterocycle selected
from the group consisting of
/ 1 /~ ~~ ~, / ~N
N N N Ni
// _N // N ~- W
/ a , // 1
N~ > Nw ~N /
R N N N Y /Z
s
-- _-
and
H
-N~
H2
N
R4 N
or
NH
SUBSTITUTE SHEET (RULE 2B)


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W is S, 0,, NH, or CH2;
Y is N or CH;
Z is N or CH;
R9 and RS are independently hydrogen, C1-C6alkyl,
aryl, C,-C6alkylaryl, C (0) 0 (C1-C6alkyl) ,
C (0) N (C1-Csalkyl ) 2, or_ C1-C6alkylCOR-,;
R, is hydrogen, C,-C6alkyl, pyrrolidinyl,
piperidinyl, homoproline, or proline;
D is C (0) R6, CHZNHSOZ (C,-Csalkyl) , or C1-
C6alkyl (OH) ;
R6 is NHS, NH (C~-Csalkyl ) , NH (C1-
C6alkylidenyl) OCH3, NH (C,-C6alkyl) aryl, N (C1-C6alkyl) 2, N (C~-
C6alkyl ) ( aryl ) , N (C~--C6alkyl ) (C1-C6alkyl aryl ) , 0 (C1-C6alkyl ) ,
piperidinyl or optionally substituted piperidinyl,
pyrrolidinyl or optionally substituted pyrrolidinyl,
pyrrolinyl or optionally substituted pyrrolinyl,
morpholino, hexameth.yleneimino, heptamethyleneimino,
quinolinyl, 2,4-dihydroquinolinyl, isoquinolinyl, 2,4-
dihydroisoquinolinyl, an amino acid selected from the
group consisting of proline, homoproline, glycine,
alanine, valine, leucine, isoleucine, tyrosine,
tryptophan, phenylalanine, serine, threonine, asparagine,
glutamic acid, aspartic acid, lysine, arginine,
glutamine, histidine, cysteine, and methionine, or a
nitrogen-containing heterocycle selected from the group
consisting of
SUBSTITUTE SHEET (RULE 26)


CA 02340344 2001-02-12
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- 14 -
N N N N
C> > o
C ,
NCH3 C N
~I
I
N
CND
N N N N
C~ C~ C~ C~
S I N I
p p~ SOZCH
N
U N~ U
0 0
N~-- i
and i
E is hydrogen, C,-C6alkyl, aryl
C1-C6alkylaryl, naphthyl, or C1-C6alkylnaphthyl,
or a pharmaceutically acceptable salt or solvate thereof.
The present invention relates to compounds of Formula
I, as follows:
SUBSTITUTE SHEET (RULE 26)


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WO 00/10565 PCT/US49/03525
- 15
H
R1 N
~NH2
O O
HN
N
Y
R3
R4
wherein R1 is C:6H5CHZOCH2-, C6H5 (CHZ) 3- or indol-3-
ylmethyl; Y is pyrrolidin-1-yl, 4-C,-C6 alkylpiperidin-
1-yl or NR2R2; R2 are each independently a C, to C6
alkyl; R3 is 2-napthyl or phenyl para-substituted by W;
W is H, F, CFA, C1-Cs alkoxy or phenyl; and R4 is H or
CH3,
or a pharmaceutically acceptable salt or solvate thereof.
The present invention further relates to
pharmaceutical formulations containing compounds of
formula I, alone or in combination with other growth
hormone secretagogue compounds, and/or in combination with
suitable bone-antiresorptive agents, and the use of said
compounds and/or formulations at least for the increase in
endogenous levels of growth hormone in a mammal.
The present invention yet further relates to methods
for the treatment or prevention of a physiological condition
which may be modulated by an increase in endogenous growth
hormone, which method comprises administering to an animal
in need of said treatment an effective amount of a compound
of formula I.
The present invention additionally relates to compounds
of formula IA:
SUBSTITUTE SHEET (RULE 2B)


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- 16 -
H
N
- NHZ
/ O O
NH
Hjy
N
CH3
O H
IA
The present invE~ntion still further relates to
compounds of formula TB:
H
N
NHZ
/ 0 O
NH
N
N ~ ~ CHs
O CH3
IB
The present invention additionally relates to
compounds of formula Ia':
SUBSTITUTE SHEET (RULE 26)


CA 02340344 2001-02-12
WO 00/10565 PCT/US99/03525
- 17 -
H
N
NHBoc
0 0
NH
N
EtO
-E
0
Ia'
wherein E is as defined above.
Also provided are compounds of formula ZZ and ZZZ
useful as chiral intermediates in the preparation of
compounds of formula I:
N02 N02
N N
E 0 0 E".. 0 O
_ N~0 N
/, O
.-
ZZ ZZZ
wherein E is as defined above.
The present invention still further relates to
processes for the prE:paration of compounds of formula I.
The terms and abbreviations used in the instant
examples have their normal meanings unless otherwise
designated. For example "°C" refers to degrees Celsius;
SUBSTITUTE SHEET (RULE 26)


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- 18 -
"N" refers to normal or normality; "mmol" refers to
millimole or millimoles; "g" refers to gram or grams; "ml"
means milliliter or milliliters; "M" refers to molar or
molarity; "MS" refer°s to mass spectrometry; "FDMS" refers
to field desorption mass spectrometry; "W" refers to
ultraviolet spectro~~copy; "IR" refers to infrared
spectroscopy; and "NMR" refers to nuclear magnetic
resonance spectroscopy.
As used herein, the term "C1-C6 alkyl" refers to
straight or branched, monovalent, saturated aliphatic
chains of 1 to 6 carbon atoms and includes, but is not
limited to, methyl, ethyl, propyl, isopropyl, butyl,
isobutyl, t-butyl, pentyl, isopentyl, and hexyl. The term
"C1-C6 alkyl" includes within its definition the term "C1-
Cq alkyl".
As used herein, the term "cycloalkyl" refers to
cyclized chains of 1 to 6 carbon atoms and includes, but is
not limited to, cyclopropyl, cyclobutyl, cyclopentyl, and
cyclohexyl.
"Halo" represents chloro, fluoro, bromo or iodo.
"C1-C6 alkoxy" represents a straight or branched alkyl
chain having from one to six carbon atoms attached to an
oxygen atom. Typical C1-C6 alkoxy groups include methoxy,
ethoxy, propoxy, isopropoxy, butoxy, t-butoxy, pentoxy and
the like. The term '"C1-C6 alkoxy" includes within its
definition the term '"C1-Cq alkoxy".
"C2-C6 alkanoyl" represents a straight or branched
alkyl chain having from one to five carbon atoms attached
through a carbonyl moiety. Typical C2-C6 alkanoyl groups
include ethanoyl (also referred to as acetyl), propanoyl,
isopropanoyl, butanoyl, t-butanoyl, pentanoyl, hexanoyl,
and the like.
"C1-C6 alkylidenyl" refers to a straight or branched,
divalent, saturated aliphatic chain of one to six carbon
atoms and includes, but is not limited to, methylenyl,
ethylenyl, propylenyl_, isopropylenyl, butylenyl,
SUBSTITUTE SHEET (RULE 26)


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- 19 -
isobutylenyl, t-butylenyl, pentylenyl, isopentylenyl,
hexylenyl, and the like.
The term "aryl." represents an aromatic ring or rings
including phenyl, napthyl, biphenyl, and aromatic residues
of 5 to 7-membered rings with 1 to 4 heteroatoms (a
"heteroaryl"), all of which may be optionally substituted
with one or more substituents, including
C1-C6 alkyl, -OC1-C6 alkyl, -OCF3, amide, NHamide,
carboxamide, sulfonamide, NHsulfonamide, imide, hydroxy,
carboxy, nitro, chloro, fluoro, tri(chloro or
fluoro)methyl, cyano, and the like. The aromatic ring may
be attached at any carbon atom or heteroatom which affords
a stable structure. 3,4-methylenedioxyphenyl is included
here.
The term "heterocycle" represents a stable 5- to 7-
membered monocyclic or 7- to 10-membered bicyclic
heterocyclic ring which is saturated or unsaturated and
which consists of carbon atoms and from 1 to 4 heteroatoms
selected from the group consisting of nitrogen, oxygen or
sulfur, and wherein ithe nitrogen and sulfur heteroatoms may
optionally be oxidized, and the nitrogen heteroatom may
optionally be quaternized and including a bicyclic group in
which any of the abo~~e-defined heterocyclic rings is fused
to a benzene ring. '.Che heterocyclic ring may be attached
at any heteroatom or carbon atom which affords a stable
structure, and may beg optionally substituted with one or
more substituents se:Lected from the group consisting of C1-
C6 alkyl, -OC1-C6 alkyl, hydroxy, nitro, chloro, fluoro, or
tri(chloro or fluoro)methyl, and the like.
The term "carbox y-protecting group" as used herein
refers to substituent:s of the carboxy group commonly
employed to block or protect the carboxy functionality while
reacting other functional groups on the compound. Examples
of such protecting groups include methyl, ethyl, p-
nitrobenzyl, p-methyl.benzyl, p-methoxybenzyl, 3,4-
dimethoxybenzyl, 2,4-~dimethoxybenzyl, 2,4,6-
SUBSTITUTE SHEET (RULE 26)


CA 02340344 2001-02-12
WO 00110565 PCT/US99/035Z5
- 20 -
trimethoxybenzyl, 2,4,6-trimethylbenzyl, pentamethylbenzyl,
3,4-methylene-dioxybenzyl, benzhydryl, 4,4'-dimethoxy-
benzhydryl, 2,2',4,4'-tetramethoxybenzhydryl, t-butyl, t-
amyl, trityl, 9-methoxytrityl, 4,4'-dimethoxytrityl, 4, 4',
4"-trimethoxytrityl, 2-phenylprop-2-yl, trimethylsilyl, t-
butyldimethylsilyl, phenacyl, 2,2,2-trichloroethyl, 2-(di(n-
butyl)methylsilyl)ethyl, p-toluenesulfonylethyl, 4-
nitrobenzylsulfonylethyl, allyl, cinnamyl, 1-
(trimethylsilylmeth;yl)prop-1-en-3-yl, and the like.
A preferred carboxy-protecting group for the practice of the
present invention is methyl or ethyl. Further examples of
these groups may be found in E. Haslam, supra, at Chapter 5,
and T . W. Greene, et a1. , supra, at Chapter 5 .
The term "amino-protecting group" as used herein
refers to substituents of the amino group commonly
employed to block or protect the amino functionality
while reacting other- functional groups on the compound.
Examples of such amino-protecting groups include formyl,
trityl, phthalimido, trichloroacetyl, chloroacetyl,
bromoacetyl, iodoacetyl, and urethane-type blocking
groups such as benzyloxycarbonyl,
4-phenylbenzyloxycarbonyl, 2-methylbenzyloxycarbonyl,
4-methoxybenzyloxycarbonyl, 4-fluorobenzyloxycarbonyl,
9-chlorobenzyloxycarbonyl, 3-chlorobenzyloxycarbonyl,
2-chlorobenzyloxycarbonyl, 2,4-dichlorobenzyloxycarbonyl,
4-bromobenzyloxycarbonyl, 3-bromobenzyloxycarbonyl,
4-nitrobenzyloxycarbonyl, 4-cyanobenzyloxycarbonyl,
n-butoxycarbonyl, (N'Boc) t-butoxycarbonyl,
1,1-diphenyleth-1-yloxycarbonyl,
1,1-diphenylprop-1-yloxycarbonyl,
2-phenylprop-2-yloxycarbonyl,
2-(p-toluyl)-prop-2-yloxycarbonyl,
cyclopentanyloxycarbonyl, 1-methylcyclopentanyloxycarbonyl,
cyclohexanyloxycarbonyl, 1-methylcyclohexanyloxycarbonyl,
2-methylcyclohexanyloxycarbonyl,
2-(4-toluylsulfonyl)-ethoxycarbonyl,
SUBSTITUTE SHEET (RULE 26)


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- 21 -
2-(methylsulfonyl)ethoxycarbonyl,
2-(triphenylphosphino)-ethoxycarbonyl,
fluorenylmethoxy-ca~__~bonyl (FMOC),
2-(trimethylsilyl)ei=hoxycarbonyl, allyloxycarbonyl,
1-(trimethylsilylmet=hyl)prop-1-enyloxycarbonyl,
5-benzisoxalylmetho~;ycarbonyl,
4-acetoxybenzyloxycarbonyl;
2, 2, 2-trichloroetho~s:ycarbonyl,
2-ethynyl-2-propoxyc:arbonyl, cyclopropylmethoxycarbonyl,
4-(decyloxy)benzyloxycarbonyl, isobornyloxycarbonyl,
1-piperidyloxycarbonyl, and the like;
benzoylmethylsulfonyl group, 2-nitrophenylsulfenyl,
diphenylphosphine oxide and like amino-protecting groups.
The amino-protecting group employed is usually not
critical so long as the derivatized amino group is stable
to the condition of ,subsequent reactions on other
positions of the intermediate molecule, and may be
selectively removed at the appropriate point without
disrupting the remainder of the molecule including any
other amino-protecting groups. A preferred
amino-protecting group for the practice of the present
invention is t-butoxycarbonyl (NBoc). Further examples
of groups referred to by the above terms are described by
E. Haslam, Protective' Groups in Organic Chemistry,
(J.G.W. McOmie, ed., 1973), at Chapter 2; and T.W. Greene
and P.G.M. Wuts, Prot:ecti ve Groups in Organi c Synthesis
(1991), at Chapter 7.
The term "leaving group" (Q) refers to a group of atoms
that is displaced from a carbon atom by the attack of a
nucleophile in a nucleophilic substitution reaction.
Suitable leaving groups include bromo, chloro, and iodo,
benzenesulfonyloxy, methanesulfonyloxy, and
toluenesulfonyloxy. 'The term "leaving group" (Q) includes
activating groups.
The term "activating group" as used herein refers a
leaving group which, when taken with the carbonyl (-C=0)
SUBSTITUTE SHEET (RULE 26)


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- 22 -
group to which it is attached, is more likely to take part
in an acylation reaction than would be the case if the
group were not present, as in the free acid. Such
activating groups are well-known to those skilled in the
art and may be, for example, succinimidoxy, phthalimidoxy,
benzotriazolyloxy, azido, or -0-CO-(Cq-C~ alkyl).
The compounds used in the method of the present
invention may have one or more asymmetric centers. As a
consequence of these chiral centers, the compounds of the
present invention occur as racemates, mixtures of
enantiomers and as individual enantiomers, as well as
diastereomers and mixtures of diastereomers. All
asymmetric forms, individual isomers and combinations
thereof, are within the scope of the present invention.
The terms "R" and "S" are used herein as commonly
used in organic chemistry to denote specific configuration
of a chiral center. The term "R" (rectus) refers to that
configuration of a c:hiral center with a clockwise
relationship of group priorities (highest to second
lowest) when viewed along the bond toward the lowest
priority group. They term "S" (sinister) refers to that
configuration of a c:hiral center with a counterclockwise
relationship of group priorities (highest to second
lowest) when viewed along the bond toward the lowest
priority group. The priority of groups is based upon
their atomic number (in order of decreasing atomic
number). A partial list of priorities and a discussion of
stereochemistry is contained in Nomenclature of Organic
Compounds: Principles and Practice, (J.H. Fletcher, et
al., eds., 1974) at pages 103-120.
In addition to the (R)-(S) system, the older D-L
system is also used in this document to denote absolute
configuration, especially with reference to amino acids.
In this system, a Fischer projection formula is oriented
so that the number 1 carbon of the main chain is at the
top. The prefix "D" is used to represent the absolute
SUBSTITUTE SHEET (RULE 26)


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- 23 -
configuration of the isomer in which the functional
(determining) group is on the right side of the carbon
atom at the chiral center and "L", that of the isomer in
which it is on the :Left.
In order to prs~ferentially prepare one optical isomer
over its enantiomer, a number of routes are available. As
an example, a mixture of enantiomers may be prepared, and
then the two enantiomers may be separated. A commonly
employed method for the resolution of the racemic mixture
(or mixture of enant:iomers) into the individual enantiomers
is to first convert the enantiomers to diastereomers by way
of forming a salt with an optically active acid or base.
These diastereomers may then be separated using differential
solubility, fractional crystallization, chromatography, or
the like. Further details regarding resolution of
enantiomeric mixtures may be found in J. Jacques, et al.,
Enantiomers, Racemates, and Resolutions, (1991).
Preferred compounds of the present invention are
compounds of formula I wherein:
A is
\ C~ ~ \ \
N or
B is
CH3
~~CH3
NHZ
SUBSTITUTE SHEET (RULE 26~

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WO 00/10565 PCT/US99/03525
- 24 -
J is H;
G is H;
X is NH;
V is
/~ /\ /
N
N N N~
I ~ or
E is
~ ~ ~ OMe
~~CH3 / ~ \ / OMe
\ \ ~ \
/ ~ ~ / ~ /
OCH3 CF3
\ ~ ~ \
/ F ~ O~\
D is -C (0) R6, where R6 is 1-pyrrolidinyl,
1-piperidinyl, 4-methyl-1-piperidinyl, N,N-dimethyl,
0 O
N ~ or N ~ ~ F
L is H or CH3;
SUBSTITUTE SHEET (RULE 26)


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- 25 -
or a pharmaceutically acceptable salt or solvate thereof.
A preferred compound includes a compound of formula Id
provided below:
Me02C
Id
Also preferred are compounds of formula IA and IB
provided hereinabove.
During any of the following synthetic sequences it may
be necessary or desirable to protect sensitive or reactive
groups on any of the molecules concerned. This may be
achieved by employing conventional protecting groups as
described, supra.
The compounds of the present invention may be prepared
by a number of routes, many of which are known to those of
skill in the art. The particular order of steps to be
employed in the synthesis of compounds of formula I is
dependent upon the compound to be synthesized, the starting
material employed, and the relative lability of the various
substituted moieties. Examples of such synthetic routes may
be found in Schemes I through IV provided below, as well as
in the Examples.
One synthetic route to compounds of the present
invention is provided in Scheme I below. The compounds of
formula IV' and IV are' commercially available, or may be
prepared using techniques known in the art. A compound of
formula IV may be prepared from a compound of formula IV'
SUBSTITUTE SHEET (RULE 26)


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- 26 -
through an intermediate acid chloride prepared by standard
methods using thionyl chloride or oxalyl chloride.
Treatment of the resulting acid chloride with a bromine
source, such as N-bromosuccinimide, followed by quenching of
the acid chloride with ethanol, results in compounds of
formula IV. It is to be understood that the bromine group
on the compound of :formula IV may in fact be any suitable
leaving group (Q), i3a defined herein. This preparation is
provided below in Scheme IA.
Scheme IA
HO'/~ Et0 Br
~R 1. thionyl chloride
p R
2. N~-bromosuccinimide
O
3 . Et:OH
IV
wherein R is representative of E as defined in a compound of
formula I above.
The starting material further includes compounds of
formula V, which are commercially available, or may be
routinely synthesized using techniques readily known in the
art. Compounds of formula IV may be coupled with a compound
of formula V (4-nitroimidazole) by methods known in the art
to generate a compound of formula IIb'. Suitable agents to
be employed in the coupling of these compounds include the
treatment of a compound of formula IV with an organic or
inorganic base, followed by reaction with the bromo compound
of formula IV. Standard organic bases include
trialkylamines, potassium hexamethyldisilazide, lithium
hexamethyldisilazide, lithium diisopropylamide, potassium
carbonate, and the like. Preferred for the practice of the
present invention is sodium hydride or potassium carbonate
in dimethylformamide. A compound of formula IIb' is then
deprotected to provide a compound of formula IIb, using
SUBSTITUTE SHEET (RULE 26)


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- 27 -
lithium hydroxide, although other deprotecting reagents may
be employed in this reaction. Such deprotecting agents
include standard saponification reagents such as sodium
hydroxide, potassitun hydroxide, and lithium hydroxide.
Substantially pure (R) enantiomers of compounds of
formula IIb may alga be synthesized by methods provided in
U. S. 5, 344, 937 and 5, 380, 866, the disclosures of which are
herein incorporated. by reference.
A compound of formula IIb is then converted to the
corresponding amide under appropriate conditions with a
compound of formula VI to generate a compound of formula
IIa. In general, a:midation of primary or secondary amines
of formula VI may be accomplished by a number of methods
known in the art in which activation of the acid to form a
better leaving group is the initial step. Suitable
activating agents for this are also known in the art and
include dicyclohexycarbodiimide (DCC), 1-(3-
dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC)
with hydroxybenzotr~_azole (HOBT), oxalyl chloride, thionyl
chloride, PyBOP~ (benzotriazol-1-yl-
oxytripyrrolidinephasphonium hexafluorophosphate), and the
like. Preferred for the practice of the present invention
is hydroxybenzotriaz;ole (HOBT). The nitro group on the
resulting compound of formula IIa may then be reduced to an
amino group using any suitable means, employing a suitable
reducing agent. Preferred for the practice of the present
invention is a catalytic reduction employing hydrogen and 5~
palladium on carbon. A compound of formula II is produced
by this reduction reaction.
The preferred reaction temperature range employed in
these reactions is between -40 and 150 °C, and the most
preferred range is between 10 and 40 °C. These reactions
may be conveniently carried out in situ, without isolation
of the particular compound after its preparation.
Examples of these reactions are provided below in
Scheme I.
SUBSTITUTE SHEET (RULE 26)


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- 28 -
Scheme I
O2N 02N
N
N
Br H N
Et0
V Et
R
R
O
NaH, DMF O
IV
IIb'
LiOH
02N 02N
R1 R2NH ~ N
VI
N
RZR1N EDC, HOBT, HO
R or oxalyl R
0 chloride O
IIa IIb
HZ~ 5$ Pd-C
H"N
G
N
RZR1N
R
O
II
wherein R is representative of E as previously defined, and
R2R1N is R6 as previously defined.
;iUBSTITUTE SHEET (RULE 26)


CA 02340344 2001-02-12
WO 00110565 PCT/US99/03525
- 29 -
A second portion of the overall synthesis of compounds
of formula I is provided in Scheme II below.
Representative starting material for this synthesis is a
compound of formula IIIb', which is a chemically-protected
form of the amino acid 0-serine. By chemically-protected it
is meant that both 'the amino- and carboxy- functional groups
have been suitably protected in order to facilitate further
reactions with this molecule. Such protection reactions are
known to those of s)cill in the art, and may be applied to
other suitable starting materials. Intermediates of formula
IIIb' are commercia7_ly available, or may be prepared by
standard syntheses of amino acids. Such syntheses are well
known to persons of ordinary skill in the art and are
described, for example, in Chemistry and Biochemistry of
Amino Acids, (G. C. C:hapman ed., 1985). The protected amino
group may be specifically deprotected using trifluoroacetic
acid and methylene chloride to allow for further reactions
with this amino funcaional group. This deprotection
reaction results in a compound of formula IIIb.
A compound of formula IIIb may then be N-acylated with
an amino-protected compound of formula X to produce a
compound of formula IIIa'. Suitable activating agents for
this N-acylation reaction are known in the art and include
DCC, HOBT, EDC, and oxalyl chloride. Preferred for the
practice of the present invention is HOBT. Compounds of
formula X are commercially available, or are readily
prepared from suitable available starting materials. The
protected carboxy group on the compound of formula IIIa' is
then selectively dep:rotected, typically using lithium
hydroxide, to generate a compound of formula III. Compounds
of formula III in which the starting material IIIb' is 2-
Nboc-amino-pentanoic acid methyl ester may also be prepared
by the route described in Scheme II.
A compound of formula III is then coupled with a
compound prepared from the reduction of IIb' with hydrogen
and a palladium cata7lyst employing a coupling reaction to
.SUBSTITUTE SHEET (RULE 26)


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generate a compound of formula Ia. Again, typical reagents
for this N-acylation are known in the art, and include DCC
and HOBT, which is the preferred method of coupling employed
in the practice of the present invention. A compound of
formula Ia is then selectively deprotected at the carboxy
group, coupled at this site with a compound of formula VI,
and then further de:protected at the amino group to generate
a compound of formu7.a Ia. Suitable agents for these
deprotection and coupling reactions are discussed, infra,
and are known in th.e art. Compounds of formula Ia are
encompassed by formula I, and axe pharmaceutically active.
The preferred reaction temperature range employed in
these reactions is between -40 and 150 °C, and the most
preferred range is between 10 and 40 °C. These reactions
may be conveniently carried out in situ, without isolation
of the particular compound after its preparation.
Alternatively, compounds of formula IIa can be coupled
with compounds of formula III to provide intermediates which
can be deprotected to give compounds of formula Ia.
Representative reactions are provided below in Scheme
II.
SUBSTITUTE SHEET (RULE 26)


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- 31 -
Scheme II
\ NHBoc NHz
O T FA \ O
i
CHZCl
COZMe 2 COZMe
IIIb' IIIb
EDC HOZC\ 'NHBoc
HOB '/~\T
X
H
\ N
0 NHBoc
/ 0 O
OMe
IIIa'
LiOH
H
\ N
0 NHBoc
/ O 0
OH
III
SUBSTITUTE SHEET (RULE 26)


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Scheme II, continued
IIb'
DCC, HOBT
H
0 N
NHBoc
~i 0 0
NH
N
Et0
R
O
Ia'
1. LiOH
2. RIRzNH VI
3. TFA
H
N
0 NH2
/ O 0
NH
N
RZR1N
R
O
Ia
.SUBSTITUTE SHEET (RULE 26)


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- 33 -
wherein R is E as previously defined, and RZR_N is Rs as
previously defined..
An alternativE~ synthetic scheme is provided in Scheme
III below. A compound of formula VII
(5-nitrobenzimidazole) is commercially available, or may be
conveniently prepared using reactions known in the art. A
compound of formula VII is coupled with a compound of
formula IV in an alkylation reaction, using coupling agents
as discussed, infra. A compound of formula VIII' is
produced in which t:he carboxy functional group is protected.
This protecting group is then removed as previously
discussed, typically using lithium hydroxide, followed by
coupling with a compound of formula XII. The nitro group on
the resulting compound of formula VIII is then reduced,
followed by coupling with a compound of formula III. The
resulting compound of formula Ib' is then deprotected to
provide a compound of formula Ib. Compounds of formula Ib
are encompassed by formula I, and are pharmaceutically
active. These reactions are provided below in Scheme III.
SUBSTITUTE SHEET (RULE 26)


CA 02340344 2001-02-12
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Scheme III
NOZ NOZ
r _
I~, \ ; 1. LiOH
N N
H NaH
DMF Et0 \ Ph 2.
VII NH
O
VI I I ' C OZMe
N02
r
1 . H2, 5 ~ Pd-C
N
v
N p, h
2.
O
N~
COzMe ° // NHBoc
O O
OH
VIII
XII
III
H / H
N N
~ NHBoc I \ \O NHZ
/ 0 O / O O
NH TFA NH
r r
Dl N
N ~ N
Ph Ph
COZMe O C02Me 0
Ib' Ib
SUBSTITUTE SHEET (RULE 2B)


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A still further representative synthesis of compounds
of formula I is prc>vided below in Scheme IV. Starting
materials of formula IX (3-amino-nitrobenzene) are
commercially available. Initially, a compound of formula IX
is coupled with a compound of formula IV by means discussed
previously. The resulting compound of formula XI' is then
deprotected, followed by coupling with a compound of formula
XII to provide a compound of formula XI. A compound of
formula XI is then reduced and further coupled in an N-
acylation reaction with a compound of formula III. The
resulting compound of formula Ic' is then deprotected to
result in a compound of formula Ic. Conditions for these
reactions have been discussed previously. Compounds of
formula Ic are encompassed by formula I, and are
pharmaceutically active.
SUBSTITUTE SHEET (RULE 28)


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Scheme IV
o2
NOz
IV
LiOH
---- \ __
NH2 N<3H NH
DMF Et0 \ Ph 2.
IX NH
O
XI , COZMe
NOz XI I
\ ~ 1. H2, 5 $ Pd-C
NH
2.
O H \/
COZMe I ~ O [[[[~~'N
O O NHBoc
XI OH
III
H
N H
~ NHBoc ~ ~ O N NHZ
/ O~ p / O O
NH NH
T FA
NH NH
N N
Ph Ph
COZMe O CO,,Me O
Ic' Ic
SUBSTITUTE SHEET (RULE 2B)


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In addition t:o the Schemes described hereinabove, an
enantiospecific protocol for the preparation of the
compounds of formula I may be employed. Typically, a
synthetic reaction design which maintains the chiral center
present in the starting material in a desired orientation is
chosen. The preferred reaction schemes are those that
generally produce compounds in which greater than 95 percent
of the product is i:he desired enantiomer. In Scheme V
below, R-substituted phenyl is representative of the E
substituents as provided in compounds of formula I above.
.SUBSTITUTE SHEET (RULE 26~

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WO 00/10565 PCTNS991035Z5
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Scheme V
OzN~N
OEt Br OEt
O -----_,.
O
R I OEt
R
I II R III
OzN
1. LiOH OzN N OzN N
Net N O 2.(COCI~
m ---~.,. ~N O N O
OEt 3. Li' ~ /~NYO + ~~~~0
NY° R o ~! o
~o
IV
V
N ~N 02N
V LiOH ~ ~I ~/ I N
N O t. oxa~y
O
2. NR2
~H ( ~
~~:'~~~NRz
R
R
VII VIII
I NHz N
X
BocHN O ~ OH
VIII ----~ N ~N'~
Pd on carbon ~'"'~~O p~ O
f
~z DCC/HOBt
R
VITIa
BoCHI
O X
1. tritlua~oacarc acid N
2. HCI
Hz~
w
~ X
,SUBSTITUTE SHEET (RULE 26)


CA 02340344 2001-02-12
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The following discussion is directed to the reactions
provided in Scheme V. Specifically, the reactions of
compounds of formula I, II, and III are as provided in the
discussion of Scheme I hereinabove.
A compound of formula IV may be prepared by the
alkylation of a compound of formula III by standard methods
using a base, such as sodium hydride, followed by treatment
with an electrophile, such as methyl iodide. Preferred
bases for this reaction include sodium-, lithium-, or
potassium hexamethyldisilazide, lithium diisopropylamide,
and sodium hydride. Preferred methylating agents include
methyl halides or any methyl group with a suitable
substituted leaving group such as toslyate, mesylate, and
the like.
A compound of i:ormula V may be prepared by hydrolysis
of a compound of formula IV using standard saponfication
conditions known in the art. Suitable reagents for this
transformation include sodium hydroxide or lithium
hydroxide. The resulting carboxylic acid may be converted
into the acid chloride by standard methods using thionyl
chloride or, preferably, oxalyl chloride. The acid chloride
may then be reacted with the lithium salt of a chiral
auxiliary, such as (4R, 55)-(+)-9-methyl-5-phenyl-2-
oxazolidinone, to provide compounds of formula V and VI,
which are readily se~aarable by silica gel chromatography.
A compound of formula VII may be prepared by the
removal of the chira:L auxiliary under basic conditions, such
as lithium hydroxide.. Other reagents known in the art for
removing oxazolidinone-type chiral auxiliaries may be used
for this transformation. These include lithium
hydroxide/hydroperoxi.de conditions, reduction/oxidation
protocols, alkyl sulfur displacements, and transaminations.
A compound of formula VIII may be prepared from a
compound of formula VII by standard methods known in the
SUBSTITUTE SHEET (RULE 26)


CA 02340344 2001-02-12
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art. Formation of tlhe acid chloride using oxalyl or thionyl
chloride followed by reaction with a suitable substituted
amine (NRz) provide compounds of formula VIII.
A compound of :Formula IX may be prepared by the
reduction of a compound of formula VIII using hydrogen with
palladium on carbon,. Other methods known in the art which
may be employed for the reduction of the nitro group include
the use of tin(II)chloride, iron in an acidic solution,
ferrous sulfate and aqueous alkali, activated alumina, and
sodium sulfite. The resulting 4-amino imidazole compound of
formula VIIa is them reacted directly with the appropriate
dipeptide acid (a cc>mpound of formula IIX) under standard
peptide coupling conditions involving formation of the
active ester of the dipeptide followed by reaction with
amine VIIa. Conditions suitable for amide formation include
DCC, EDC, with HOBT. A compound of formula IIX may be
prepared from the methyl ester of unnatural D-amino acids
such as D-benzyloxyserine, D-tryptophan, and D-2-amino-5-
phenyl-pentanoic acid and the like which are known in the
art. Standard coupling protocols involving formation of the
active ester of the amino acid using DCC/HOBt followed by
reaction with N-Boc.-aminoisobutyric acid provide dipeptide
acids of formula ITX.
The Boc protecting group of a compound of formula IX
may be removed under standard acidic conditions such as
hydrochloric acid in acetic acid or ethyl acetate,
trifluoroacetic acid, tetramethyliodosilane, aluminum
chloride, sulfuric acid in dioxane, and methanesulfonic
acid.
An additional method of preparing diastereomeric
compounds of formula I involves the use of a chromatographic
column which employs a chiral phase, An example of such a
preparation may be found in Examples Part 6 as provided
hereinbelow.
SUBSTITUTE SHEET (RULE 26)


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Preferred for the practice of the present invention are
those compounds of formula I wherein the indicated
stereochemistry is (R,R) at the two chiral centers. An
example of this preferred stereochemistry is provided by
compounds of formula IA and IB as provided hereinabove.
Two additional Schemes for providing chiral
intermediates are provided hereinbelow as Schemes VA and VB.
As described in Scheme VA, optically pure aryl glycine amino
acids may be protected at the amino position by reaction
with a suitable pros=ecting group, such as Boc. Reaction of
the Boc protected intermediate with a standard methylating
agent, such as methyl iodide, may provide the corresponding
phenolic methyl ether. The carboxamide may be prepared by
coupling with an amine, such as dimethylamine, pyrrolidine,
or 4-methyl piperidi_ne, using standard coupling techniques.
Preferred coupling agents for the invention are diethy
cyanophosphorane (DE;CP), triethylamine and the amine at O°C.
The Boc protecting group may be removed under standard
acidic conditions, with trifluoroacetic acid being
preferred. The desired 4-nitroimidazole compounds can be
prepared by reaction of the free amine with 1, 4-
dinitroimidazole to give optically pure compounds, as
determined by chiral HPLC. Such chiral intermediates can be
processed as described in Schemes I and II to provide
diastereomerically pure products. For example, the chiral
nitroimidazoles described in Scheme VA or VB may be reduced
under standard conditions, such as hydrogenation with a
palladium catalyst, to provide the corresponding chiral
amino intermediate I:C. Such intermediates may be
subsequently coupled with compounds of type III as
previously described to provide a chiral intermediate which
can be deprotected to give diastereomerically pure compounds
of formula Ia.
SUBSTITUTE SHEET (RULE 26)


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Scheme VA
CHIRAh SYNTHESIS of D-Phenylglycine Imidazole
H= NHBoc NBOc
OH ~BoC70 y ~pH NaH
OH


NaOH Mel
p 0


HO / p
HO


MBO


la)p~-I56(1N HCI1 [a)p~-1201MeOH)


(aloe-134(MeOH!


DECP,tt3H.C'C
O=N p=N HN ~
N)iz NHBoc
m NOT \ ~TFA , ~ \
p NaHCO~ p
O
Me0 MeOH,H20 Me0 Me0
[alo~-258.2(MeOH)
lalp~-165.5(MoOH)
Chlcal HPLC>97\ee
S
SUBSTITUTE SHEET (RULE 26)


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Scheme YB
Chiral Synthesis of L-Phenylglycine Imidazale
NHBoc
OH NHBoc
THF N
HN ----
p DCC, HOBt
/ O
[a]D= +144(EtOH) [a]p=+95.9(MeOH)
OZN
O.,N TFA
N
NHz
N02 ~ N
[a]p +177.6(MeOH) N
MeOH,HzO ~ O
[a)D=+19.7(1N HC1)
Chiral HPLC>97$ee
An additional approach and corresponding synthetic
scheme for the preparation of compounds of the instant
invention is provided below in Scheme VI:
SUBSTITUTE SHEET (RULE 26)

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Scheme VI
/ . o / I o
\ I O ~ ~~ \ I OMe 110H \ OH ~ ~N I
NH dio~ ~ NH ~2 HCJ
NHZ ~flq C~ Me
~ ~ R
Me' I 'NHBoc goc~ BocHN Me
N
R=
ci
CDMT= N' 'N
NNM= N~Nhylrtarpholine
MeGr 'N' _OMe ~ N~,
~2
/ I O I~ / / O /
\ ~ \ \~ ''~ \I
N TFA an'sole
NH H ~ NH
O ~2 Oc~(~ R
R
Nle
~2TFA BocHN Me
HZN Nle
N N
U N~ R= U
R=
Ma
Mle
:SUBSTITUTE SHEET (RULE 26)


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Pharmaceutically active compounds of formula I include
at least compounds of formula IA, IB, Id, and Ia' as
described herein.
Compounds of formula I may be conveniently screened for
growth hormone secretagogue activity. A typical assay may
employ pituitary cells established in culture, followed by a
challenge with the various compounds of formula I, and the
levels of growth hormone determined accordingly. Growth
hormone levels may be calculated using various
radioimmunoassay techniques known to those of skill in the
art. Screening of compounds for growth hormone secretagogue
activity may conveniently be scaled up for high throughput
screening.
The invention :Further encompasses methods employing the
pharmaceutically acceptable salts of the compounds defined
by formula I. Although generally neutral, a compound of
this invention can possess a sufficiently acidic, a
sufficiently basic, or both functional groups, and
accordingly react with any of a number of inorganic bases,
and inorganic and organic acids, to form a pharmaceutically
acceptable salt.
The term "pharmaceutically acceptable salt" as used
herein refers to salts of the compounds of formula I which
are substantially non-toxic to living organisms. Typical
pharmaceutically acceptable salts include those salts
prepared by reaction. of the compounds of the present
invention with a pharmaceutically acceptable mineral or
organic acid or an inorganic base. Such salts are known as
acid addition and base addition salts.
Acids commonly employed to form acid addition salts are
inorganic acids such as hydrochloric acid, hydrobromi.c acid,
hydroiodic acid, sulfuric acid, phosphoric acid, and the
like, and organic acids such as p-toluenesulfonic,
methanesulfonic acid, oxalic acid,
SUBSTITUTE SHEET (RULE 2B)


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p-bromophenylsulfon:ic acid, carbonic acid, succinic acid,
citric acid, benzoic acid, acetic acid, and the like.
Examples of such pharmaceutically acceptable salts are the
sulfate, pyrosulfate:, bisulfate, sulfite, bisulfite,
phosphate, monohydrogenphosphate, dihydrogenphosphate,
metaphosphate, pyrophosphate, chloride, bromide, iodide,
acetate, propionate, decanoate, caprylate, acrylate,
formate, isobutyrate, caproate, heptanoate, propiolate,
oxalate, malonate, :cuccinate, suberate, sebacate, fumarate,
maleate, butyne-1,4-~dioate, hexyne-1,6-dioate, benzoate,
chlorobenzoate, methylbenzoate, dinitrobenzoate,
hydroxybenzoate, met.hoxybenzoate, phthalate, sulfonate,
xylenesulfonate, phenylacetate, phenylpropionate,
phenylbutyrate, citrate, lactate, y-hydroxybutyrate,
glycollate, tartrate, methanesulfonate, propanesulfonate,
naphthalene-1-sulfonate, naphthalene-2-sulfonate, mandelate,
mesylate, and the like. Preferred pharmaceutically
acceptable acid addition salts are those formed with mineral
acids such as hydrochloric acid and hydrobromic acid, and
those formed with arganic acids such as malefic acid and
methanesulfonic acid.
Salts of amine groups may also comprise quaternary
ammonium salts in which the amino nitrogen carries a
suitable organic group such as an alkyl, alkenyl, alkynyl,
or aralkyl moiety.
Base addition salts include those derived from
inorganic bases, such as ammonium or alkali or alkaline
earth metal hydroxides, carbonates, bicarbonates, and the
like. Such bases useful in preparing the salts of this
invention thus include sodium hydroxide, potassium
hydroxide, ammonium hydroxide, potassium carbonate, sodium
carbonate, sodium bicarbonate, potassium bicarbonate,
calcium hydroxide, calcium carbonate, and the like. The
potassium and sodium salt forms are particularly preferred.
It should be recognized that the particular counterion
forming a part of any salt of this invention is not of a
SUBSTITUTE SHEET (RULE 26)


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critical nature, so long as the salt as a whole is
pharmacologically acceptable and as long as the counterion
does not contribute undesired qualities to the salt as a
whole.
This invention further encompasses methods employing
pharmaceutically acceptable solvates of the compounds of
Formula I. Many of the formula I compounds can combine with
solvents such as water, methanol, ethanol and acetonitrile
to form pharmaceutically acceptable solvates such as the
corresponding hydrate, methanolate, ethanolate and
acetonitrilate.
This invention also encompasses methods employing the
pharmaceutically acceptable prodrugs of the compounds of
formula I. A prodrug is a drug which has been chemically
modified and may be biologically inactive at its site of
action, but which may be degraded or modified by one or more
enzymatic or other ~:n vivo processes to the parent bioactive
form. This prodrug should have a different pharmacokinetic
profile than the parent, enabling easier absorption across
the mucosal epithelium, better salt formation or solubility,
or improved systemic stability (an increase in plasma half-
life, for example).
Typically, such chemical modifications include:
1) ester or amide derivatives which may be cleaved by
esterases or lipases;
2) peptides which may be recognized by specific or
nonspecific proteases; or
3) derivatives that accumulate at a site of action
through membrane selection of a prodrug form or a modified
prodrug form; or any combination of 1 to 3, supra.
Conventional procedures for the selection and preparation of
suitable prodrug derivatives are described, for example, in
H, Bundgaard, Design of Prodrugs, (1985).
As used herein, the term "effective amount" means an
amount of compound o:E the instant invention which is capable
of inhibiting, alleviating, ameliorating, treating, o:r
SUBSTITUTE SHEET (RULE 26)


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preventing further symptoms in mammals, including humans,
which may be due to decreased levels of endogenous growth
hormone.
By "pharmaceutically acceptable formulation" it is
meant that the carrier, diluent, excipients and salt must be
compatible with the active ingredient (a compound of formula
I) of the formulation, and not be deleterious to the
recipient thereof. Pharmaceutical formulations can be
prepared by procedures known in the art. For example, the
compounds of this invention can be formulated with common
excipients, diluents, or carriers, and formed into tablets,
capsules, and the like. Examples of excipients, diluents,
and carriers that are suitable for such formulations include
the following: fillers and extenders such as starch,
sugars, mannitol, and silicic derivatives; binding agents
such as carboxymethyl cellulose and other cellulose
derivatives, alginates, gelatin, and polyvinyl pyrrolidone;
moisturizing agents ouch as glycerol; disintegrating agents
such as agar agar, calcium carbonate. and sodium
bicarbonate; agents for retarding dissolution such as
paraffin; resorption accelerators such as quaternary
ammonium compounds; ;surface active agents such as cetyl
alcohol, glycerol monostearate; adsorptive carriers such as
kaolin and bentonite,; and lubricants such as talc, calcium
and magnesium stearai=a and solid polyethylene glycols. Final
pharmaceutical forms may be: pills, tablets, powders,
lozenges, syrups, aerosols, saches, cachets, elixirs,
suspensions, emulsions, ointments, suppositories, sterile
injectable solutions,. or sterile packaged powders, and the
like, depending on the type of excipient used.
Additionally, the compounds of this invention are well
suited to formulation as sustained release dosage forms.
The formulations can also be so constituted that
they release the act~~ve ingredient only or preferably in a
particular part of the intestinal tract, possibly over a
period of time. Such formulations would involve coatings,
SUBSTITUTE SHEET (RULE 26)


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envelopes, or protective matrices which may be made from
polymeric substances or waxes.
The particular dosage of a compound required to treat,
inhibit, or prevent the symptoms and/or disease of
congestive heart failure in a mammal, including humans,
according to this invention will depend upon the particular
disease, symptoms, and severity. Dosage, routes of
administration, and frequency of dosing is best decided by
the attending physician. Generally, accepted and effective
doses will be from l5mg to 1000mg, and more typically from
l5mg to 80mg. Such dosages will be administered to a
patient in need of treatment from one to three times each
day or as often as needed for efficacy.
In addition, the growth hormone secretagogue compounds
as disclosed herein may be administered to a patient in need
of treatment in combination with other growth hormone
secretagogues known :in the art, and/or with a suitable bone
anti-resorptive agent or agents for the prevention or
treatment of osteoporosis and/or loss of muscle strength.
Said suitable bone anti-resorptive agents include selective
estrogen receptor modulators, bisphophonates, calcitonin,
and hormone replacement therapeutic agents. Additionally,
PTH may be administered in combination with said growth
hormone secretagogues. Said combination therapy may be
administered concomii:antly or sequentially.
Suitable dosing ranges of compounds of formula I
include 0.01 ~.g/kg/day to 50 mg/kg/day.
The present invention also relates to methods for the
modulation of cardiac: function which comprise the
administration of a compound of Formula I.
The present invention further relates to methods for
the treatment or prevention of congestive heart failure by
administering, to an animal in need thereof, an effective
amount of a compound of Formula I.
SUBSTITUTE SHEET (RULE 26)


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The present invention additionally relates to
pharmaceutical formulations containing a growth hormone
secretagogue alone or in combination with additional
therapeutic agents useful for the treatment or prevention of
congestive heart failure.
The use of growth hormone secretagogue compounds, for
the modulation of cardiac function and for the treatment or
prevention of congestive heart failure, are described in
copending U.S. Patent Application Serial No. 09/137,255,
filed August 19, 1998, titled "Treatment of Congestive Heart
Failure With Growth Hormone Secretagogues", the teachings of
which are incorporated herein in their entirety by
reference.
The particular dosage of a compound required to treat,
inhibit, or prevent the symptoms and/or disease of
congestive heart failure in a mammal, including humans,
according to this invention will depend upon the particular
disease, symptoms, and severity. Dosage, routes of
administration, and frequency of dosing is best decided by
the attending physic:i_an. Generally, accepted and effective
doses will be from lSmg to 1000mg, and more typically from
l5mg to 80mg. Such dosages will be administered to a
patient in need of treatment from one to three times each
day or as often as needed for efficacy.
Representative pharmaceutical formulations containing
compounds of formula I are provided below. The formulations
which follow are given for purposes of illustration and are
not intended to be limiting in any way. The total active
ingredients in such formulations comprises from 0.1~ to
99.9$ by weight of the formulation. The term "active
ingredient" means a compound of Formula I.
SUBSTITUTE SHEET (RULE 26)


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Formulation 1
Hard gelatin capsules containing the following
ingredients are prepared:
Quantity
Ingredient (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 2
A tablet formula is prepared using the ingredients
below:
Ingredient Quantity
(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 3
A dry powder inhaler formulation is prepared containing
the following components:
Ingredient Weight ~
Active Ingredient 5
Lactose 95
The active mixture is mixed with the lactose and the
mixture is added to a, dry powder inhaling appliance.
SUBSTITUTE SHEET (RULE 26)


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Formulation 4
Tablets, each containing 30 mg of active ingredient,
are prepared as follows:
Quantity
Ingredient


(mg/tablet)


Active Ingredient 30
0


.
mg


Starch 45.0 mg


Microcrystalline cellulose 35
0


.
mg


Polyvinylpyrrolidone


(as 10~ solution in water) 4.0 mg


Sodium carboxymethy.l starch 4
5


.
mg


Magnesium stearate 0
5


.
mg


Talc 1.0 mg


Total 120 mg



The active ingi:edient, starch and cellulose are passed
through a No. 20 me:~h U.S. sieve and mixed thoroughly. The
solution of polyvinylpyrrolidone is mixed with the resultant
powders, which are then passed through a 16 mesh U.S. sieve.
The granules so produced are dried at 50-60°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 120 mg.
Formulation 5
Capsules, each containing 40 mg of medicament are made
as follows:
Quantity
Ingredient (mg/capsule)
Active Ingredient 40.0 mg
Starch 109.0 mg
Magnesium stearate 1.0 mg
Total 150.0 mg
SUBSTITUTE SHEET (RULE 26)


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The active ingredient, cellulose, 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 6
Suppositories, each containing 25 mg of active
ingredient are made as follows:
Ingredient Amount
Active Ingredient 25 mg
Saturated fatty acid glycerides 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.() g capacity and allowed to cool.
Formulation 7
Suspensions, each containing 50 mg of medicament per
5.0 ml dose are made as follows:
Ingredient Amount


Active Ingredient 50.0 mg


Xanthan gum 4.0 mg


Sodium carboxymethyl cellulose (11$)


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 medicament, 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
SUBSTITUTE SHEET (RULE 26)


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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 $
Capsules, each containing 15 mg of medicament, are made
as follows:
Quantity
Ingredient


(mg/capsule)


Active Ingredient 15.0 mg


Starch 407.0 mg


Magnesium stearate 3.0 mg


Total 425.0 my



The active ingi:edient, cellulose, starch, and magnesium
stearate are blended, passed through a No. 20 mesh U.S.
sieve, and filled into hard gelatin capsules in 425 mg
quantities.
Formulation 9
An intravenous formulation may be prepared as follows:
Ingredient Quantity
Active Ingredient 250.0 mg
Isotonic saline 1000 ml
Formulation 10
A topical formulation may be prepared as follows:
Ingredient
Quantity
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
SUBSTITUTE SHEET (RULE 26)


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stirring is continued until dispersed. The mixture is then
cooled until solid.
Formulation 11
Sublingual or buccal tablets, each containing 10 mg of
active ingredient, may be prepared as follows:
Quantity
Ingredient Per Tablet
Active Ingredient 10.0 mg
Glycerol 210.5 mg
Water 143.0 mg
Sodium Citrate 4.5 mg
Polyvinyl Alcohol 26.5 mg
Polyvinylpyrrolidone~ 15.5 mg
Total 410.0 mg
The glycerol, water, sodium citrate, polyvinyl alcohol,
and polyvinylpyrrolidone are admixed together by continuous
stirring and maintaining the temperature at about 90°C.
When the polymers have gone into solution, the solution is
cooled to about 50-55°C and the medicament is slowly
admixed. The homogenous mixture is poured into forms made
of an inert material. to produce a drug-containing diffusion
matrix having a thickness of about 2-4 mm. This diffusion
matrix is then cut t:o form individual tablets having the
appropriate size.
Another formulation employed in the methods of the
present invention employs transdermal delivery devices or
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 transderm.al patches for the delivery of
pharmaceutical agents is well known in the art. See, for
example, U.S. Patent 5,023,252, the disclosure of which is
herein incorporated by reference. Such patches may be
constructed for continuous, pulsatile, or on demand delivery
of pharmaceutical agents.
SUBSTITUTE SHEET (RULE 26)


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Frequently, it will be desirable or necessary to
introduce the pharmaceutical composition to the brain,
either directly or indirectly. Direct technirnies usually
involve placement of a drug delivery catheter into the
host's ventricular aystem 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 5,011,472, the disclosure
of which is herein incorporated by reference.
Indirect techn~'_ques, which are generally preferred,
usually involve formulating the compositions to provide for
drug latentiation by the conversion of hydrophilic drugs
into lipid-soluble drugs or prodrugs. Latent~ation is
generally achieved through blocking of the hydroxy,
carbonyl, sulfate, a.nd primary amine groups present on the
drug to render the drug more lipid soluble and amenable to
transportation acrass 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.
The following Examples and Preparations are
illustrative of the processes employed in the synthesis of
the compounds of the present invention. As would be
understood by persons skilled in the art, other synthetic
schemes may be employed to prepare the compounds of the
instant invention.
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EXAMPLES PART 1
Preparation 1
O NHBOC
CO Me
2
To a solution of boc-(OBz)-D-Ser-OH (25.0 g, 84.7 mmol)
stirring in anhydrous N,N-dimethylformamide (500 mL) at room
temperature was added sodium bicarbonate (14.2 g, 169 mmol)
followed by methyl iodide (26.4 mL, 424 mmol). After' 18 h,
the reaction mixture was concentrated to approximately 100
mL. Ethyl acetate was added and the mixture washed with
aqueous sodium bicarbonate and brine. The organic extract
was dried and concentrated to give the desired compound (25
g, 96$) as a light yellow oil: 1H NMR (300 MHz, CDC13) d
1.45 (s, 9H), 3.70 (m, 1H), 3.75 (s, 3H), 3.85 (m, 1H), 4.50
(m, 3H), 7.30 (m, 5H); MS (FD) m/e 310; Anal. calc'd for
C16H23N05: C, 62.12; H, 7.49; N, 4.53. Found: C, 62.31; H,
7.49; N, 4.43.
Preparation 2
o~r~2
COZMe
To a solution of a compound of Preparation 1 (5.0 g, 16
mmol) stirring in dichloromethane (25 mL) and anisole (1 mL)
at 0 oC was added trifluoroacetic acid. After 4 h at room
SUBSTITUTE SHEET (RULE 26)


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temperature, saturai:ed sodium bicarbonate solution was added
and the mixture extracted with ethyl acetate. The combined
organic extracts were washed with brine, dried over sodium
sufate, and concentrated. The crude product was used in the
next step without further purification.
Preparation 3
H
'~ O ~ N NHBoc
Me0 2C O
To a solution of a compound of Preparation 2 (65.4
mmol), boc-a-aminoisobutyric acid (13.2 g, 65.4 mmol), 1-
hydroxybenzotriazole~ (8.8 g, 65.4 mmol), and N,N-
diisopropylethylamine (22.8 mL, 130.7 mmol) stirring in
dichloromethane ( 500 mL) at 0 °C was added 1- ( 3-
dimethylaminopropyl)-3-ethylcarbodiimide (12.3 g, 71.9
mmol). After 18 h, ethyl acetate and saturated ammonium
chloride were added and the mixture extracted with ammonium
chloride, sodium bicarbonate, and brine. The organic
extracts were dried over sodium sulfate and concentated.
Purification by silica gel chromatography (25$ ethyl
acetate/hexanes) yielded the desired compound (21.6 g', 83$)
as a white solid: 1H NMR (300 MHz, CDC13) d 1.39 (s, 9H),
1. 48 (s, 6H) , 3. 62 (dd, J = 3.4, 9. 1 Hz, 1H) , 3. 70 (s, 3H) ,
3.85 (dd, J = 3.4, 9.1 Hz, 1H), 4.48 (dd, J = 12.5, 22.7 Hz,
2H), 4.75 (m, 1H), 4.92 (s, 1H), 7.11 (d, J = 8.6 Hz, 1H),
7.35 (m, 5H); M5 (FD) m/e 395; Anal. calc'd for C2pH30N2C6:
C, 60.90; H, 7.67; N, 7.10. Found: C, 61.02; H, 7.78; N,
7.10.
SUBSTITUTE SHEET (RULE 28)


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Preparation 4
H
O~ N NHBoc
H02C O
To a solution of a compound of Preparation 3 (5.30 g,
13.4) stirring in dioxane (100 mL)/water (50 mL) at room
temperature was addE~d lithium hydroxide (2.80 g, 67.3 mmol).
After 18 h, water was added and the solution concentrated.
The resulting mixture was extracted with diethyl ether.
Sodium chloride was added to the aqueous layer and the pH
adjusted to 3.5 witl-i 1 N HC1. The resulting mixture was
extracted with ethyl. acetate and the combined organic
extracts dried over sodium sulfate then concentrated to
yield the title compound (4.40 g, 86~) as a white foam: 1H
NMR (300 MHz, CDC13) d 1.39 (s, 9H) , 1 .45 (s, 3H) , 1.47 (s,
3H), 3.68 (m, 1H), 3.95 (m, 1H), 4.54 (s, 2H), 4.70 (m, 1H),
5.51 (bs, 1H), 7.18 (d, J = 9.1 Hz, 1H), 7.25 (m, 5H), 9.90
(bs, 1H); MS (FD) m/e 381; Anal. calc'd for C19H28N206: C,
59.99; H, 7.42; N, 7.36. Found: C, 59.74; H, 7.26; N, 7.30.
Preparation 5
Br
OEt
O
To a solution o:E a-bromophenylacetic acid (100 g, 466
mmol) stirring in absolute ethanol (500 mL) at room
temperature was added p-toluenesulfonic acid monohydrate (10
g, 53 mmol). This solution was heated to reflux and, after
SUBSTITUTE SHEET (RULE 26j


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8 h, concentrated to dryness. The resulting residue was
dissolved in ethyl acetate, washed with saturated aqueous
sodium bicarbonate, brine, dried over sodium sulfate,
filtered, and concentrated to yield 77 g (68 $) of the
desired product as an orange oil: 1H-NMR is consistent with
structure; MS (FD) 241.9, 243.9.
Preparation 6
NOZ
N
Et0
I
O
To a slurry of sodium hydride (13.6 g of a 60$
dispersion in mineral oil, 341 mmol) stirring in N,N-
dimethylformamide (240 mL) was carefully added 4-
nitroimidazole (38.6 g, 391 mmol) such that the temperature
during the addition was maintained below 40°C. This
resulting slurry wasp stirred for 1 h and then cooled to 5°C.
To this mixture was slowly added BX8-MEZ-148 (76 g, 310
mmol) at a rate such that the reaction temperature was
maintained below 20°C. After 4 h, the reaction was
concentrated and subsequently extracted with ethyl acetate.
The combined organic extracts were filtered, washed with
water, brine, dried over sodium sulfate, filtered and
concentrated. The resulting residue was purified by silica
gel chromatography (methanol/chloroform gradient) to yield
the 60.1 g (70$) oi= the desired product as a white solid:
1H-NMR is consistent with structure; MS (FD) 275 (M+); Anal.
Calc'd. for: C, 56.73; H, 4.73; N, 15.27. Found: C, 56.48;
H, 4.78; N, 15.08.
SU8ST1TUTE SHEET (RULE 26)


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Preparation 7
Bc
To a suspension of 5$ Pd/C (0.85 g) and a compound of
Preparation 6 (2.13 g, 7.21 mmol) stirring in dioxane (50
mL) at room temperature was added hydrogen (g) (35 psi) on a
Parr apparatus. After 4 h, the mixture was purged with
nitrogen, celite added, and the solution filtered through a
pad of celite. To the resulting~filtrate, under nitrogen
atmosphere, was added a compound of Preparation 4 (2.74 g,
7.21 mmol), 1-hydro:xybenzotriazole (0.97 g, 7.21 mmol), N,N-
diisopropylethylamine (2.5 mL, 14.4 mmol), and 1-(3-
dimethylaminapropyl)-3-ethylcarbodiimide (1.36 g, 7.93
mmol). After 18 hours, ethyl acetate was added and the
mixture washed with saturated aqueous ammonium chloride,
saturated aqueous sodium bicarbonate, and brine. The
organic extract was dried over sodium sulfate and
concentrated. Purification by silica gel chromatography (5$
methanol/dichloromet=hane) yielded the title compound (1.25
g, 29 $ ) as a yellow foam: 1H NMR ( 300 MHz, CDC13 ) d 1 . 30
(t, J = 6.9 Hz, 3H) , 1.40 (s, 9H) , 1.42 (s, 3H) , 1.51 (s,
3H) , 3 . 60 (dd, J = 5.1, 9. 7 Hz, 1H) , 4 . 05 (m, 1H) , 4 .28 (m,
2H) , 4 .54 (dd, J = 7.4. 08, 26.3 Hz, 2H) , 4 . 62 (m, 1H) , 5. 08
(bs, 1H) , 5. 82 (s, 1H) , 7.12 (d, J = 11 . 5 Hz, IH) , 7.35 (m,
12H), 9.75 (bs, 1H); MS (FD) m/e 607; Anal. calc'd for
C32H41N5~7: C, 63.2'9; H, 6.80; N, 11.52. Found: C, 63.07;
H, 6.81; N, 11.74.
SUBSTITUTE SHEET (RULE 26)


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Preparation 8
, O
BocHN
N -
O N
O H O
HO
To a solution of a compound of Preparation 7 (5.3 g,
8.75) stirring in dioxane (50 mL)/water (25 mL) at room
temperature was added lithium hydroxide (0.73 g, 17.50
mmol). After 20 min, water was added and the reaction
concentrated to approximately 30 mL. The resulting mixture
was extracted with diethyl ether and the aqueous layer
saturated with sodium chloride then adjusted to pH 3.5 with
1 N HC1. The mixture was extracted with ethyl acetate and
the combined organic extracts dried over sodium sulfate and
concentrated to yield the title compound (4.90 g, 97$) as a
light orange foam: 1H NMR (300 MHz, CDC13) d ; MS (FD) m/e
Anal, calc'd for . C, ; H,; N, . Found: C, ; H, ; N, .
Preparation 9
N~
_ N
N~ O
H N
SUBSTITUTE SHEET (RULE 2B)


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To a solution of a compound of Preparation 8 (2.09 g,
3.61 mmol), pyrrolidine (0.30 mL, 3.61 mmol), and 1-
hydroxybenzotriazole (0.59 g, 3.97 mmol) stirring in
anhydrous DMF (50 mL) at 0 °C was added 1,3-dicyclohexyl
carbodiimide (0.82 ~g, 3.97 mmol). After 18 hours at room
temperature, the reaction was concentrated, dissolved in
dichloromethane, filtered, and concentrated. Purification
by silica gel chromatography (5$ methanol/dichloromethane)
yielded the title compound (1.74 g, 76~) as a light orange
solid: 1H NMR (300 MHz, CDC13) d 1.41 (s, 9H), I.43 (s,
3H) , 1 . 52 (s, 3H) , 2 . 88 (m, 4H) , 3. 42 (m, 1H) , 3. 50 (m, 4H) ,
4.08 (m, 1H), 4.55 (dd, J = 14.9, 27.4 Hz, 2H), 4.70 (m,
1H), 4.96 (d, J = 4..0 Hz, 1H), 5.86 (s, 1H), 7.15 (d, J =
6. 9 Hz, 1H) , 7. 35 (rn, 12H) , 9.28 (bs, 1H) ; MS (FD) m/e 632;
Anal. calc'd for C3~~H44N606: C, 64.54; H, 7.01; N, 13.28.
Found: C, 63.48; H, 6.95; N, 12.19.
Example 1
O
H
H N'~I N N~
2 H
~~ O
To a solution of a compound of Preparation 9 (1.00 g,
1.58 mmol) and anisole (0.3 mL) stirring in anhydrous
dichloromethane (12 mL) at 0 °C was added trifluoroacetic
acid (3 mL) and the reaction mixture warmed to room
temperature. After 4 h, the dichloromethane was removed in
SUBSTITUTE SHEET (RULE 26)


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vacuo and excess diethyl ether added. After 20 min, the
reaction mixture wars filtered to yield the title compound
(1.02 g, 85$) as a white solid: IH NMR (300 MHz, CDC13) d
1. 60 (s, 6H) , 1. 90 (m, 4H) , 3. 08 (m, iH) , 3. 58 (m, 3H) , 3. 88
(m, 2H) , 4 . 52 (m, 2H1 , 4 . 72 (m, 1H) , 6.10 (m, 2H) , 7 .25 (m,
6H) , 7 . 46 (m, 5H) , 7 .70 (m 1H) , 8 . 00 (m, 1H) , 8 . 40 (m, 1H) ,
11.15 (m, 1H); MS (1?D) m/e 532 (M-2TFA); Anal. calc'd for
C33H38F6N6~8: C, 52.10; H, 5.03; N, 11.05. Found: C, 51.54;
H, 5.25; N, 11.21.
Preparation 10
O
F:tO
Br
To a slurry of d,l-a-amino-4-phenylbutyric acid (20.0
g, 111 mmol) stirring in 3N sulfuric acid (200 mL) at 0°C
was added finely ground potassium bromide (48 g, 403 mmol).
This slurry was cooled to -10°C, then a solution of sodium
nitrite (11.0 g, 160 mmol in water (75 mL)) was added
dropwise. The resulting solution was stirred for 4 h while
slowly warming to ambient temperature. The resulting
precipitate was filtered to give 20.0 g of a yellow solid.
To a solution of the yellow solid (18.8 g, 80 mmol) in
absolute ethanol (400 mL) was added p-toluenesulfonic acid
monohydrate (4.6 g, 24 mmol). This solution was refluxed
for 4 h, filtered and concentrated. The resulting residue
was purified by silica gel chromatography (ethyl
acetate/hexanes gradient) to give 7.2 g (24$) of the desired
product as a clear oil. 1H-NMR is consistent with structure;
MS (FD) 269, 27
SUBSTITUTE SHEET (RULE 26)


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Preparation 11
N02
N
N
EtO _
v
O
To a slurry of sodium hydride (1.0 g of a 60$
dispersion in mineral oil, 24 mmol) stirring in N,N-
dimethylformamide (200 mL) at ambient temperature was
carefully added a solution of 4-nitroimidazole (5.7 g, 20
mmol). This mixture was cooled to 0 °C and a solution of a
compound of Preparation 10 (15.2 g, 60 mmol) in N,N-
dimethylformamide (:10 mL) was added. After 16 h, the
mixture was slowly caarmed to ambient temperature,
concentrated, and the resulting residue extracted with
chloroform. The connbined organic extracts were washed with
water, brine, dried over sodium sulfate, filtered and
concentrated. The resulting residue was purified by silica
gel chromatography (chloroform) to give 5.0 g (82$) of the
desired product as a. clear oil. 1H-NMR is consistent with
structure; MS (FD) 303 (M+); Anal. Calc'd for: C, 59.40; H,
5.65; N, 13.85. Found: C, 59.73; H, 5.71; N, 13.40.
SUBSTITUTE SHEET (RULE 26)


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Preparation I2
N02
N
N
H:O
p
To a solution o:f a compound of Preparation 11 (4.24 g,
14 mmol) stirring in tetrahydrofuran (30 mL) and ethanol (30
mL) at room temperai~ure was added 2N NaOH (35 mL, 70 mmol) .
After 1 h, this mixture was treated with 5N HC1 until pH =
2.5. Ethyl acetate ~;30 mL) and water (30 mL) were added and
the resulting solution was extracted with ethyl acetate.
The combined organic: extracts were washed with brine, dried
over sodium sulfate, filtered and concentrated to give 3.8 g
(98$) of the desired product as a yellow oil: 1H-NMR is
consistent with stru.cture~ MS (FD) 276 (M+),
SUBSTITUTE SHEET (RULE 2B)


CA 02340344 2001-02-12
wo oon os6s pcT~smo3s2s
-67-
Preparation 13
NOZ
N
N
MeO .C', . O
:.
To a solution of a compound of Preparation 12 (3.8 g,
14 mmol ) , 1-proline methylester ( 1. 8 g, 14 mmol ) and 1-
hydroxybenzotriazole hydrate (2.1 g, 15 mmol) stirring in
N,N-dimethylformamide (150 mL) at room temperature was added
1, 3-dicyclohexylcarbodiimide (3.2 g, 15.4 mmol) . After 16
h, the mixture was concentrated and the resulting residue
partitioned between ethyl acetate and water. The combined
organic extracts were washed with water, brine, dried over
sodium sulfate, and concentrated. The resulting orange oil
was purified by silica gel chromatography
(methanol/chloroform gradients) to give 3.8 g (70$) of the
desired product as a yellow oil: 1H-NMR is consistent with
structure; MS (FD) 386.2 (M+).
SUBSTITUTE SHEET (RULE 26)


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Preparation 14
O N
NHBoc
O O
HN
N
N
N
_ v
MfeO 2 C .
To a slurry of 10~ Pd/C in dioxane (10 mL) was added a
solution of a compound of Preparation 13 (2.4 g, 6.2 mmol)
in dioxane (100 mL). The mixture was then treated with
hydrogen gas (40 psi) on a Parr apparatus. After 5 h, an
amount of 10$ Pd/C (0.5 g) in dioxane (10 mL) was added.
The mixture was hydt-ogenated for 4 h then carefully filtered
through celite. To the resulting filtrate was added a
compound of Preparation 4 (2.4 g, 6.2 mmol) , 1-
hydroxybenzotriazole~ hydrate (0.92 g, 6.8 mmol), followed by
1,3-dicyclohexylcarbodiimide (1.4 g, 6.8 mmol). After 16 h,
the reaction was concentrated and the resulting residue
extracted with ethyl acetate. The combined organic extracts
were washed with saturated aqueous sodium bicarbonate,
brine, dried over sodium sulfate, filtered, and
concentrated. Purification silica gel chromatography
(methanol/chloroform gradient) gave 2.2 g (50~) of the
desired product as a tan foam: 1H-NMR is consistent with
structure; MS (FD) 718.7 (M+); Anal Calc'd for: C, 63.49;
H, 7.01; N, 11.69. Found: C, 63.30; H, 6.91; N, 11.84.
SUBSTITUTE SHEET (RULE 26)


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Example 2
H
O N
O O
N
Me0 zC
To a solution of a compound of Preparation 14 (2.1 g,
3.0 mmol) stirring :in dichloromethane (25 mL) was added
trifluoroacetic acid (8 mL, 104 mmol). After one h, water
(25 mL) was added and the solution was quenched carefully
with sodium carbonate, then extracted with chloroform. The
combined organic extracts were washed with brine, dried over
sodium sulfate, filtered and concentrated. To a solution of
the resulting residue in diethyl ether (40 mL) was added a
saturated solution of HC1 in diethyl ether (40 mL). The
resulting slurry was concentrated to dryness to yield 1.6 g
(80~) of the desired. product as a tan foam: 1H-NMR is
consistent with structure; MS (FD) 618.3 (M+); Anal. Calc'd.
for: C, 57.31; H, 6.41; N, 12.15. Found: C, 57.52; H, 6.19;
N, 12.04. IR (KBr) 2954, 1743, 1656, 1559, 1496, 1453 cm 1.
SUBSTITUTE SHEET (RULE 26)


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Preparation 15
0
Et0
Br
Reaction of d,l-a-phenylalanine (20.0 g, 120 mmol),
potassium bromide (48 g, 400 mmol), sodium nitrite (ll.Og,
150 mmol), water (75 mL), 3N sulfuric acid (200 mL), p-
toluenesulfonic acid monohydrate (5.7 g, 30 mmol) and
absolute ethanol (500 mL) according to Preparation 10 gave
18.0 g (70~) of the desired product as a colorless oil: 1H-
NMR is consistent w:Lth structure; MS (FD) 256, 258.
Preparation 16
NO 2
N
Et0
0
Reaction of a compound of Preparation 15 (15.22 g, 60
mmol), sodium hydride (2.84 g of a 60~ dispersion in mineral
oil, 72 mmol), 4-nitroimidazole (8.1 g, 72 mmol) in N,N-
dimethylformamide (400 mL) according to Preparation 11 gave
9.5 g (55$) of the desired product as a yellow foam: 1H-NMR
is consistent with structure; MS (FD) 289.1 (M+); Anal.
Calc'd. for: C, 58.13; H, 5.23; N, 14.53. Found: C, 58.40;
H, 5.17; N 14.24.
SUBSTITUTE SHEET (RULE 26)


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Preparation 17
NOz
N
HO
O
s
Reaction of a compound of Preparation 16 (3.3 g, 12.0
mmmol ) , 2N NaOH ( 30 mL, 60 mmol ) in ethyl acetate ( 30
mL)/ethanol (30 mL) according Preparation 12 gave 2.85 g
(90$) of the desired. product as a white solid: 1H-NMR is
consistent with structure; MS (FD) 262 (M+); Anal. Calc'd.
for: C, 55.17; H, 4.24; N, 16.09. Found: C, 55.14; H, 4.24;
N, 15.94.
Preparation 18
NOZ
N
N
v v
MeO~ ZC' O
Reaction of a compound of Preparation 17 (2.8 g, 11.C
mmol ) , 1-proline methylester ( 1. 4 g, 11 . 0 mmol ) , 1-
hydroxybenzotriazole hydrate (1.63 g, 12.1 mmol), and 1,3-
dicyclohexylcarbodiimide (2.5 g, 12.1 mmol) in N,N-
dimethylformamide (150 mL according to Preparation 13 gave
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3.2 g (70.4$) of th.e desired product as a white solid: 1H-
NMR is consistent with structure; MS (FD) 372 (M+).
Preparation 19
NHBoc
O O
HN 2HC1
N
N
v v
M~,,O yC O
Reaction of a compound of Preparation 18 (0.6 g, 1.6
mmol), 5$ Pd/C (0.66 g) in ethyl acetate (50 mL), ethanol
(50 mL) and dichloromethane (4 mL), a compound of
Preparation 9 (0.46 g, 1.2 mmol), 1-hydroxybenzotriazole
hydrate (0.18 g, 1.3 mmol) and 1,3-dicyclohexylcarbodiimide
(0.27 g, 1.3 mmol) i:n N,N-dimethylformamide (100 mL)
according to Preparation 14 gave 0.29 g (34$) of the desired
product as a tan foam: 1H-NMR is consistent with structure;
MS (FD) 704.5 (M+) ,
.SUBSTITUTE SHEET (RULE 26)


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Example 3
H
O N
'\~~~~ 2
0 0
HN 2HC1
N
<~ N I
N
Me0 ZC
Reaction of a compound of Preparation 19 (0.23 g, 0.33
mmol) , trifluoroacet:ic acid (4.0 mL, 24 mmol) in
dichloromethane (12 mL), followed by treatment with
HC1/ethyl acetate solution (40 mL), according to Example 2
gave 0.17 g (77$) of the desired product as a white foam:
1H-NMR is consistent with structure; MS (FD) 604 (M+); Anal.
Calc'd for: C, 56.72; H, 6.25; N, 12.40. Found: C, 56.53;
H, 6.31; N, 12.19. IR (KBr) 2931.09, 1743.64, 1653.48,
1533.67, 1453.73 (cm 1) .
Preparation 20
N02
N
Et0
0
Reaction of ethylbromoacetate (4. 9 mL, 44 mmol) , 4-
nitroimidazole (5.00 g, 44 mmol) and potassium carbonate
(12.2 g, 88 mmol) at ambient temperature in N,N-
SUBSTITUTE SHEET (RULE 26)


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dimethylformamide (50 mL) according to Preparation 3 from
Examples Part 2A gave 7.77 g (88~) of the desired product as
an orange solid: 'H-NMR was consistent with structure; M5
(FD) 199 (M+); Anal. Calc'd for: C, 42.21; H, 4.55; N,
21.10. Found: C, X12.51; H, 4.66; N, 21.24.
Preparation 21
NO 2
HO
O
Reaction of a ~~ompound of Preparation 20 (2.00 g, 10.0
mmol) and 2N NaOH (:30 mL, 60 mmol) in tetrahydrofuran (5 mL)
and ethanol (5 mL) according Preparation 12 gave 1.3 g (76$)
of the desired product as a tan solid which is carried on
without further purification.
Preparation 22
NO 2
N
N
_.
Me0 Z C O
Reaction of a compound of Preparation 21 (1.20 g, 7.0
mmol), 1-proline methylester hydrochloride (1.27 g, 8.4
mmol), 1-hydroxybenzotriazole hydrate (1.04 g, 8.4 mmol),
triethylamine (1.95 mL, 14.0 mmol) and 1,3-
dicyclohexylcarbodii:mide ( 1. 6 g, 8 . 4 mmol ) in N, N-
dimethylformamide according to Preparation 13 gave 0.6 g
SUBSTITUTE SHEET (RULE 26)


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(30~) of the desired compound as a tan semi-solid: 1H-NMR
is consistent with :structure: MS (FD) 282 (M+) .
Preparation 23
H
O N
NHBoc
O O
HN
N
N I
Me0 C.
2
Hydrogenation of a compound of Preparation 22 (0.47 mg,
1.7 mmol) and 5 $ Pd-C (0.15 g) in ethyl acetate (20
mL)/ethanol (20 mL) followed by treatment with 1-
hydroxybenzotriazole hydrate (225 mg, 1.7 mmol), 1,3-
dicyclohexylcarbodiimide (340 mg, 1.7 mmol) and 368979 (633
mg, 1.7 mmol) according to Preparation 14 gave 0.95 g (39~)
of the desired product: 1H-NMR is consistent with structure;
MS (FD) 614 (M+) .
SUBSTITUTE SHEET (RULE 26)


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Example 4
0
NH 2
O O
HN
N
N II
lVleO Z C O
Reaction of a compound of Preparation 23 (0.40 g, 0.65
mmol) and trifluoroacetic acid (5 mL, 64 mmol) in
dichloromethane (20 mL) according to Example 2 gave 0.22 g
(67$) of the desired product as an off-white solid: 1H-NMR
is consistent with :>tructure; MS (FD) 514 (M+); Anal. Calc'd
for: C, 58.35; H, 5.66; N, 16.33. Found: C, 58.25; H, 6.40;
N, 16.16.
Preparation 24
N02
E1
Reaction of 5-nitroindole (3.0 g, 18.5 mmol), a-
bromophenylacetic acid ethylester (4.5 g, 18.5 mmol), and
sodium hydride (0.8 ct, 20 mmol, 60$ dispersion in mineral
oil) in N,N-dimethylfarmamide (75 mL) according to
Preparation 1 gave 3.9 g (65$) of the desired product: 1H-
SUBSTITUTE SHEET (RULE 26)


CA 02340344 2001-02-12
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NMR is consistent with structure; MS (FD) 324 M+; Anal.
Calc' d for: C, 66. E~6; H, 4.97; N, 8. 64 . Found: C, 66. 80; H,
5.11; N, 8.81,
Preparation 25
NO 2
N
HO
I
O
Reaction of a compound of Preparation 24 (2.0 g, 6.2
mmol) and 2N NaOH (5.0 mL, 100 mmol) in tetrahydrofuran (10
mL)/ethanol (8 mL) according to Preparation 12 gave 1.4 g
(76$) of the desired product as a yellow solid: 1H-NMR is
consistent with structure; MS (FD) 296 (M+); Anal. Calc'd
for: C, 64.86; H, 4.08; N, 9.45. Found: C, 64.60; H, 4.14;
N, 9.29.
SUBSTITUTE SHEET (RULE 26)


CA 02340344 2001-02-12
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Preparation 26
N02
N
I
N
Me0 2C.
Reaction of a compound of Preparation 25 (1.0 g, 5.7
mmol), 1-hydroxybenzotriazole hydrate (O.BS g, 6.3 mmol), 1-
proline methylester hydrochloride (1.03 g, 6.3 mmol),
triethylamine ( 1. 6 rnL, 11 . 4 mmol ) and l, 3-
dicyclohexylcarbodi~.mide ( 1. 3 g, 6 . 3 mmol ) in N, N-
dimethylformamide (25 mL) according to Preparation 13 gave
1.35 g (58g) of the desired product as yellow solid: 1H-NMR
is consistent with ~;tructure; MS (FD) 407 (M+); Anal. Calc'd
for: C, 64.86; H, 5.20; N, 10.31. Found: C, 65.20; H, 5.50;
N, 10.10.
Preparation 27
H
O N .
NHBoc
O O
HN
Me02C
SUBSTITUTE SHEET (RULE 26)


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Hydrogenation of a compound of Preparation 26 (0.41 g,
1.0 mmol) with 5 ø Pd-C (0.08 g) in ethanol (25 mL) /ethyl
acetate (25 mL) followed by treatment with 1-
hydroxybenzotriazole hydrate (0.15 g, 1.1 mmol), 1,3-
dicyclohexylcarbodi.imide (0.23 g, 1.1 mmol) and 368979 (0.42
g, 1.1 mmol) according to Preparation 14 gave 0.38 g (51$)
of the desired product: 1H-NMR is consistent with structure;
MS (FD) 739.7 (M+)
Example 5
O
IV~i2
O O
HN
Me0 2C
Reaction of a compound of Preparation 27 (0.38 g, 0.51
mmol) and trifluoroacetic acid (2 mL, 26 mmol) in
dichloromethane (10 mL) according to Example 2 gave 0.125 g
(38~) of the desired product: 1H-NMR is consistent with
structure; MS (FD) 6:39 (M+) ; Anal . Calc' d for 1 H20: C,
65.74; H, 6.59; N, 10.65. Found: C, 65.75; H, 6.42; N,
10.98.
SUBSTITUTE SHEET (RULE 28)


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Preparation 28
NO 2
~"'~ N
N
HO
To a solution ~of a compound of Preparation 6 (27 g, 98
mmol) stirring in tetrahydrofuran (60 mL) and absolute
ethanol (60 mL) at ambient temperature was added 2N NaOH
(250 mL, 500 mmol). After 3.5 h, the mixture was washed
with diethyl ether and the organic extract subsequently
washed with water. The combined aqueous extracts were
acidified and the rE:sulting mixture extracted with ethyl
acetate. The combined organic extracts were washed once
with brine, dried over sodium sulfate, filtered, and
concentrated to give' 24.2 g (75$) of the desired product as
a tan solid: 1H-NMR was consistent with structure; MS (FD)
246.9 (M+) ; Anal. Ca.l.c'd for: C, 53.44; H, 3. 67; N, 17.00.
Found: C, 53.71; H, 3.67; N, 16.83, mp = 218-221°C.
Preparation 29
N02
N
~N
J
Me0 2C O
To a slurry of <i compound of Preparation 28 (8.15 g, 33
mmol) stirring in dichloromethane (100 mL) was added oxalyl
SUBSTITUTE SHEET (RULE 26~


CA 02340344 2001-02-12
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chloride (11.5 mL, 130 mmol) and N,N-dimethylformamide (2
drops). After 90 min at ambient temperature, the mixture
was concentrated and the residue was dissolved in
dichloromethane (40 mL). The resulting solution was added a
N,N-diisopropylethylamine (6.5 mL, 360 mmol) and 1-
prolinemethylester (3.9 g, 20 mmol) in dichloromethane (4
mL). After 2 h at ambient temperature, the mixture was
extracted with ethyl acetate and the combined organic
extracts washed with water, brine, dried over sodium
sulfate, filtered, <~nd concentrated. The resulting residue
was purified by silica gel chromatography (ethyl
acetate/hexanes) to give 10.7 g (71~) of the desired product
as tan foam: 1H-NMR. is consistent with structure; Anal.
Calc'd for: C, 56.98; H, 5.06; N, 15.63. Found: C, 56.75;
H, 5.14; N, 15.44. Mp, 103-111°C.
Preparation 30
NHBoc
O
O
HN
N
Me0 2C
To a slurry of 5~ Pd/C (0.28 g) in ethyl acetate (30
mL) was added a solution of a compound of Preparation 29
(1.0 g, 2.8 mmol) in ethanol (100 mL). The mixture was
hydrogenated at 40 p;;i on a Parr apparatus. After 25 min,
additional 5o Pd/C (0.5 g) was added and the mixture
susbsequently hydrogE~nated for 45 min, then filtered through
celite and concentrai_ed. To a slurry of the resulting
SUBSTITUTE SHEET (RULE 26)


CA 02340344 2001-02-12
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residue in N,N-dimethylformamide (100 mL) was added boc-d-
benzyloxyserine (0.62 g, 2.1 mmol), 1-hydroxybenzotriazole
hydrate(0.31 g, 2.3 mmol) followed by 1,3-
dicyclohexylcarbodiimide (0.48 g, 2.3 mmol). After 48 h, the
mixture was filtered and concentrated and the residue
purified by radial chromatography (silica gel,
methanol/chloroform gradient). The resulting product was
dissolved in ethyl acetate and washed with water, dried over
sodium sulfate, filtered, and concentrated to give 0.5 g
(30$) of the desired product as a tan foam: 'H-NMR is
consistent with structure; MS (FD) 605 (M+).
Preparation 31
0 NH z
O
HN
N
N
Me0 ZC
To a solution of a compound of Preparation 30 (3.1 g,
5.1 mmol) stirring i:n methanol (200 mL) at room temperature
was added 5N HCI (51.0 mmol). After 16 h, the residue was
partitioned between ethyl acetate and water and extracted
with ethyl acetate. The combined organic extracts were
washed with brine, dried over sodium sulfate, filtered and
concentrated to give 2.1 g (81$) of the desired compound as
a tan foam: 1H-NMR is consistent with structure; MS (FD)
506 (M+); Anal. Calc'd. for: C, 64.14; H, 6.18; N, 13.85.
Found: C, 63.92, H, 6.18; N, 13.56.
SUBSTITUTE SHEET (RULE 26)


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Preparation 32
0
~1~NHBoc
O
HN
/'1N N
Me0 zC
O
To a solution of a compound of Preparation 31 (2.1 g,
4.2 mmol) stirring in N,N-dimethylformamide (200 mL) was
added Boc-a-aminoisobutyric acid (0.85 g, 4.2 mmol), 1-
hydroxybenzotriazole hydrate(0.62 g, 4.6 mmol). After 16 h,
mixture was concentrated to dryness and the resulting
residue extracted with ethyl acetate. The combined organic
extracts were washed with water, brine, dried over sodium
sulfate, filtered, and concentrated. Purification by silica
gel chromatography (methanol/chloroform) gave 2.3 g (80~) of
the desired product as a tan foam: 1H-NMR is consistent
with structure; MS (FD) 690 (M+).
Example 6
'~ O
~~NH2
0 O
HN
MeO 2C
2.5 HC1
SUBSTITUTE SHEET (RULE 2t3)


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To a solution of the compound of Preparation 32 (1.75
g, 2.5 mmol) stirring in dichloromethane (190 mL) was added
trifluoroacetic acid (63 mL, 780 mmol). After 1 h, the
mixture was poured carefully into saturated aqueous sodium
bicarbonate and extracted with ethyl acetate. The combined
organic extracts were washed with brine, dried aver sodium
sulfate, filtered, and concentrated. The residue was
dissolved in ethyl acetate (250 mL) and subsequently treated
with a saturated solution of HC1 in ethyl acetate (100 mL).
The resulting mixture was concentrated to dryness,
triturated with diethyl ether, and filtered to give 0.6 g
(38g) of the desired product as a tan solid: 1H-NMR is
consistent with structure; MS (FD) 590 (M+); Anal. Calc'd
for: C, 54.60; H, 5.92; N, 12.33. Found: C, 54.47; H, 5.72;
N, 12.16. IR (KBr) 3164, 3030, 2978, 2952, 2878, 1743,
1664, 1531, 1456, 1436, 1498, 1197, 1179 cm'1.
Preparation 33
O ZN
~~N
INIeO 2 C O
The optically enriched S-isomer was isolated by
selective crystallization (ethyl acetate/hexanes) of a
compound of Preparation 29 to give 1.3 g of the desired
isomer; 1H-NMR is consistent with structure; MS (FD) 358
(M+); Anal. Calc'd for: C, 56.98; H, 5.06; N, 15.63. Found:
C, 57.22; H, 4.87; N, 15.34. mp = 114-118°C.
Preparation 34
SUBSTITUTE SHEET (RULE 26)


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O NHBoc
O
HN
N
N
Me0 ZC O
Hydrogenation of a compound of Preparation 29 (1.0 g,
2.8 mmol) and 5o Pd/C (0.756 g) in absolute ethanol (20
mL)/ethyl acetate (20 mL), followed by treatment of the
resulting mixture with boc-d-benzyloxyserine (0.83 g, 2.8
mmol), 1-hydroxybenzotriazole hydrate (0.42 g, 3.4 mmol) and
1,3-dicyclohexylcar:bodiimide (0.64 g, 3.1 mmol according to
Preparation 1 gave 0.69 g (41~) of the desired product as a
crystalline solid. Purification by silica gel
chromatography (metlzanol/chloroform) followed by re-
crystalization from ethyl acetate: 1H-NMR is consistent
with structure; MS (FD) 605 (M+); Anal. Calc'd. for: C,
63.46; H, 6.49; N, 11.56. Found: C, 63.61; H, 6.31; N,
11.38; mp = 184-18E~°C.
Preparation 35
0 ~2
HN
Me0 -
SUBSTITUTE SHEET (RULE 26)


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Reaction of a compound of Preparation 34 (0.61 g, 1.0
mmol) and trifluoroacetic acid (1.7 mL, 22 mmol) in
dichloromethane (90 mL) according to Preparation 1 gave 0.5
g (100$) of the deaired product as a foam: 1H-NMR is
consistent with structure; MS (FD) 506 (M+); mp = 55-60°C.
Preparation 36
0 ~
~~~~NHBoc
O O
HN
N
~N N
iMeO Z C O
Reaction of a compound of Preparation 35 (0.5 g, 1
mmol), 1-hydroxybenzotriazole hydrate (0.15 g, 1.1 mmol) and
1,3-dicyclohexylcark>odiimide (0.23 g, 1.1 mmol) in N,N-
dimethylformamide (1.5 mL) according to Preparation 32 gave
0.69 g (100$) of the desired product as a foam: 1H-NMR is
consistent with structure; MS (FD) 690.2 (M+); mp = 81-84°C.
I5
Example 7
0
,..~~NH 2
0 0
HN
i
MfeO 2 C
SUBSTITUTE SHEET (RULE 2B)


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Reaction of a compound of Preparation 36 (0.595 g, 0.95
mmol) and trifluoroacetic acid (0.7 mL, 9.0 mmol) in
dichloromethane (2:5 mL) according to Preparation 1 gave 0.37
9 (75$) of the desired product as a solid: 1H-NMR was
consistent with structure; MS (FD) 590 (M+); Anal. Calc'd
for: C, 63.04; H, 6.48; N, 14.23. Found: C, 62.98; H, 6.59;
N, 14.01. Mp, 156--159°C.
Preparation 37
O ~ IVHBoc
O
HN
<~ N
Me0 2C
Reaction of a compound of Preparation 29 (2.63 g, 8.0
mmol), boc-1-benzyloxyserine (2.4 g, 8.0 mmol), 1-
hydroxybenzotriazole hydrate (1.2 g, 8.8 mmol), 1,3-
dicyclohexylcarbodiimide (1.8 g, 8.8 mmol) in N,N-
dimethylformamide (250 mL) gave 2.4 g (50$) of the desired
product as tan foam: 1H-NMR is consistent with structure;
MS (FD) 605 {M+) ,
SUBSTITUTE SHEET (RULE 26)


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Preparation 38
0~~2
O
HN
~N
l
_i
Me0 2C
Reaction of a compound of Preparation 37 (2.3 g, 3.8
mmol) , trifluoroacet:ic acid (35 mL, 45 mmol) in
dichloromethane (90 mL) gave 1.4 g (74$) of the desired
product as a tan foam: 1H-NMR is consistent with structure;
MS (FD) 506 (M+) .
Preparation 39
O~ ~ NHBoc
O=,.I O
HN
N
~N N
Me0 ZC O
Reaction of a compound of Preparation 38 (1.1 g, 2.2
mmol), boc-a-aminoiso:butyric acid (0.45 g, 2.2 mmol), 1-
hydroxybenzotriazole hydrate (0.33 g, 2.4 mmol) and 1,3-
dicyclohexylcarbodiimide (0.5 g, 2.4 mmol) in N,N-
dimethylformamide (100 mL) gave 0.84 g (55$) of the desired
SUBSTITUTE SHEET (RULE 26)


CA 02340344 2001-02-12
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product as a tan foam: 1H-NMR is consistent with structure;
MS (FD) 690 (M+) ,
Example 8
O O~ NH2
O
HN
fN " HCl
~N
Me02C
Reaction of a compound of Preparation 39 (0.7 g, 1.0
mmol) , trifluoroacetic acid (25 mL, 320 mmol) in
dichloromethane, and followed by dissolution in ethyl
acetate (100 mL) and treatment with ethyl acetate saturated
with HC1 (100 mL) y:ielded 0.29 g (44$) of the desired
compound as a white solid: 1H-NMR is consistent with
structure; MS (FD) 590 (M+); Anal. Calc'd for: C, 56.11; H,
6.08; N, 12.66. Found: C, 56.16; H, 5.92; N, 12.56. IR
(KBr) 3163.75, 3031..15, 2952.46, 2876.38, 1745.07, 1664.94,
1530.69, 1497.79, 1453.37, 1435.81, 1197.21, 1177.62,
1094.93, 747.95, 701..04 cm 1.
SUBSTITUTE SHEET (RULE 28)


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Preparation 40
N02
O~
~N
O
To a solution of a compound of Preparation 28 (1.0 g,
4.0 mmol), morpholine (0.35 mL, 4.0 mmol), 1-
hydroxybenzotriazolE~ hydrate (0.6 g, 4.4 mmol) stirring in
N,N-dimethylformamide (50 mL) at room temperature was added
1,3-dicyclohexylcarbodiimide (0.9 g, 4.4 mmol). After 16 h,
the mixture was concentrated, and the residue extracted with
ethyl acetate. The combined organic extracts were filtered,
washed with saturated aqueous sodium bicarbonate, water,
brine, dried over sodium sulfate, filtered, and
concentrated. The resulting residue was purified by silica
gel chromatography (methanol/chloroform) to give 0.75 g
(60$) of the desired product as a white foam: 'H-NMR is
consistent with structure; MS (FD) 316 (M+).
SUBSTITUTE SHEET (RULE 26)


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Preparation 41
H
/~ 0 N
~~~ NHBoc
O O
HN
N
0 N-
~N
I
To a slurry of 5g Pd/C (0.18 g) in ethyl acetate (5 mL)
was carefully added a solution of a compound of Preparation
40 (0.67 g, 2.0 mmol) in ethyl acetate (25 mL)/ethanol (25
mL). The resulting slurry was treated with hydrogen gas at
40 psi on a Parr apparatus. After 1 h, a slurry of 5$ Pd/C
(0.18 g) in ethyl acetate (10 mL) was added to this mixture,
followed by hydrogenation at 40 psi. After 1 h, the mixture
was filtered through celite and concentrated. To the
residue stirring in D1,N-dimethylformamide (100 mL) was added
a compound of Preparation 9 (0.53 g, 1.4) and 1-
hydroxybenzotriazole hydrate (0.21 g, 1.54 mmol) followed by
1,3-dicyclohexylcarbodiimide (0.32 g, 1.54 mmol). After 16
h at room temperature, the solution was concentrated and
extracted with ethyl acetate. The combined organic extracts
were washed with water, brine, dried over sodium sulfate,
filtered, and concentrated. The resulting residue was
purified by silica gel chromatography (methanol/chloroform)
to yield 0.27 g (30~) of the desired product as a tan foam:
1H-NMR is consistent with structure; MS (FD) 448 (M+).
;iUBSTITUTE SHEET (RULE 26)


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Example 9
I 0 ~'~ NH z
0 O
HN
N
~N
O
2HC1
To a solution of a compound of Preparation 41 (0.27 g,
0.42 mmol) stirring in dichloromethane (12 mL) at room
temperature was adcLed trifluoroacetic acid (4 mL, 51 mmol) .
After 1.5 h, water (40 mL) was added and the reaction
mixture quenched carefully with solid sodium bicarbonate.
The resulting mixture was extracted with ethyl acetate and
the combined organic extracts washed with brine, dried over
sodium sulfate, filtered and concentrated. The concentrate
was dissolved in ethyl acetate (40 mL) and subsequently
treated with a saturated solution of HC1 in ethyl acetate
(40 mL). After 15 min, the mixture was concentrated to give
0.14 g (54$) of the desired product as a white solid: 1H-NMR
is consistent with structure; MS (FD) 548 (M+); Anal.
Calc'd. for: C, 56.04; H, 6.16; N, 13.52. Found: C, 55.78;
H, 6.11; N, 13.27; IR (KBr) 2927, 2858.9, 1659.3, 1542.2,
1114.4 cm i.
SUBSTITUTE SHEET (RULE 26)


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Preparation 42
NOz
CI N
N
1
O
Reaction of a compound of Preparation 28 (1.0 g, 4.0
mmol), piperidine (0.4 mL, 4.0 mmol), 1-hydroxybenzotriazole
hydrate (0.6 g, 4.4 mmol) and 1,3-dicyclohexylcarbodiimide
( 0 . 9 g, 4 . 4 mmol ) .in N, N-dimethylformamide ( 50 mL) gave 0. 95
g (75$) of the desired product as a tan foam: 'H-NMR is
consistent with structure; MS (FD) 314 (M+).
Preparation 43
O
NHBoc
O O
c
Hydrogenation of a compound of Preparation 42 (0.91 g,
2.9 mmol) in ethyl acetate (50 mL)/ethanol (50 mL), 5~ Pd/C
(0.36 g) in ethyl acetate (5 mL) followed by reaction with a
compound of Preparation 4 (0.95 g, 2.5 mmol), 1-
hydroxybenzotriazole hydrate (0.37 g, 2.75 mmol), and 1,3-
SUBSTITUTE SHEET (RULE 26)


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dicyclohexylcarbodiimide (0.5? g, 2.75 mmol) gave 0.43 g
(25$) of the desired product as a tan foam: 1H-NMR is
consistent with structure; MS (FD) 646 (M+),
Example 10
H
~2
N '\~
O O
HN
N
I
N
O
2HC1
Reaction of a compound of Preparation 43 (0.38 g, 0.59
mmol) and trifluoroacetic acid (4 mL, 51 mmol) in
dichloromethane (12 mL) followed by acidification with HC1
gave 0.03 g (8.30) of the desired product as a tan solid:
1H-NMR is consistent with structure; MS (FD) 546 (M+); IR
(KBr) 3141, 2937, 2859, 1642, 1539, 1453, 1444 cm 1.
Preparation 44
O NHBoc
C~' O
HN
SUBSTITUTE SHEET (RULE 26)


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To a slurry of 5~ Pd/C (1.0 g) in ethyl acetate (25 mL)
was added a solution of a compound of Preparation 6 (8.25 g,
30 mmol) in ethyl acetate (25 mL)/absolute ethanol (25 mL).
The slurry was hydrogenated at 40 psi on a Parr apparatus.
After 75 min, a slurry of 5$ Pd/C (0.7 g) in ethyl acetate
(25 mL) was added t:o the reaction mixture. After
hydrogenation at 40 psi for 1.5 h, the mixture was filtered
through celite and concentrated. The concentrate was
dissolved in N,N-di.methylformamide (500 mL) and boc-d-
benzyloxyserine (9.0 g, 30.8 mmol), 1-hydroxybenzotriazole
hydrate (4.5 g, 33 mmol) and 1,3-dicyclohexylcarbodiimide
(6.8 g, 33 mmol) added. After 16 h at ambient temperature,
the mixture was concentrated and the residue extracted with
ethyl acetate. The combined organic extracts were washed
with water, brine, dried over sodium sulfate, filtered, and
concentrated. Purification by silica gel chromatography
(methanol /chloroform) gave 8.33 g (53~) of the desired
product as a tan solid: 1H-NMR is consistent with
structure; MS (FD) 522 (M+); Anal. Calc'd. for: C, 64.35; H,
6.56; N, 10.72. Found: C, 64.59; H, 6.83; N, 10.77.
Preparation 45
0 NH 2
O
HN
N
N
Et0
I
O
To a solution of a compound of Preparation 44 (8.1 g,
15.5 mmol) stirring at room temperature in dichloromethane
(75 mL) was added t:rifluoroacetic acid (25 mL, 320 mmol).
SUBSTITUTE SHEET (RULE 26)


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After 50 min, the mixture was carefully poured into a
saturated aqueous solution of sodium bicarbonate and
extracted with ethyl acetate. The combined organic extracts
were washed with brine, dried over sodium sulfate, filtered
and concentrated to give 6.5 g (990) of the desired product
as a tan solid. 'H-rfMR is consistent with structure; MS (FD)
422 (M+) .
Preparation 46
0
~~ NHBoc
0 O
HN
N
N
Et0
O
To a solution of a compound of Preparation 45 (6.5 g,
15.0 mmol), boc-a-aminoisobutyric acid (3.05 g, 15.0 mmol),
1-hydroxybenzotriazo:Le hydrate (2.23 g, 16.5 mmol) stirring
in N,N-dimethylformamide (400 mL) at room temperature was
added 1,3-dicyclohe:xylcarbodiimide (3.4 g, 16.5 mmol).
After 16 h, the mixture was concentrated and the resulting
residue extracted with ethyl acetate. The combined organic
extracts were washed with water, brine, dried over sodium
sulfate, filtered, and concentrated. Purification by silica
gel chromatography (methanol/chloroform) gave 6.39 g (70~)
of the desired product as a tan foam: 1H-NMR is consistent
with structure; MS (FD) 607 (M+). Anal. Calc'd. for: C,
63.25; H, 6.80; N, 11.52. Found: C, 63.36; H, 6.92; N,
11.59.
SUBSTITUTE SHEET (RULE 26)


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Preparation 47
O
NHBoc
C~~ O o
HN\
~N
,/~N~
HO
To a solution of a compound of Preparation 46 (6.04 g,
9.9 mmol) stirring in absolute ethanol
(50 mL)/tetrahydrofuran (50 mL) at room temperature was
added 1N NaOH (50 m:L, 49.5 mmol). After 30 min, the mixture
was acidified with 1N HC1 and extracted with ethyl acetate.
The combined organic extracts were washed with brine, dried
over soditun sulfate,, filtered, and concentrated to give 5.4
g (94~) of the desired product as a tan foam: 1H-NMR is
consistent with structure; MS (FD) 580 (M+); Anal. Calc'd.
for: C, 62.16; H, 6..43; N, 12.08. Found: C, 61.86; H, 6.29;
N, 12.06.
Preparation 48
H
O N
NHBoc
O O
HN
N
N
MeHN
SUBSTITUTE SHEET (RULE 26)


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To a solution of a compound of Preparation 47 (0.7 g,
1.2 mmol), N-methylamine hydrochloride (0.08 g, 1.2 mmol),
triethylamine (0..'i mL, 3.6 mmol), and 1-hydroxybenzotriazole
hydrate (0.18 g, 1..32 mmol) stirring in N,N-
dimethylformamide (50 mL) at room temperature was added 1,3-
dicyclohexylcarbod.iimide (0.27 g, 1.32 mmol). After 16 h,
the mixture was concentrated and the resulting residue
extracted with ethyl acetate. The combined organic extracts
were washed with water, brine, dried over sodium sulfate,
filtered, and concentrated. Purification by silica gel
chromatography (methanol /chloroform) gave 0.25 g (35~) of
the desired product= as a white solid: 1H-NMR is consistent
with structure; MS (FD) 592.4 (M+); Anal. Calc'd for 0.32
mol hydrate: C, 62,21; H, 6.76; N, 14.04. Found: C; 62.17;
H, 6.74; N, 14.19.
Example 11
H
~2
O N '\~
O O
HN
N
MeHN
O
2HC1
To a slurry of a compound of Preparation 48 (0.2 g,
0.34 mmol) stirring in dichloromethane (12 mL) at room
temperature was added trifluoroacetic acid (4 mL, 52 mmol).
After 2 h, additional trifluoroacetic acid (4 mL, 52 mmol)
was added and the reaction was heated to reflux. After 7 h,
the mixture was cooled to room temperature, water (40 mL)
SUBSTITUTE SHEET (RULE 26)


CA 02340344 2001-02-12
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added, followed excess solid sodium bicarbonate. The
mixture was extracted with ethyl acetate and the combined
organic extracts washed with brine, dried over sodium
sulfate, filtered and concentrate. The resulting crude
product was dissolved in ethyl acetate (40 mL) and a
saturated solution of HC1 in diethyl ether was added C40
mL). After 15 min, this slurry was concentrated to give
0.13 g (68$) of the desired product as a white solid: 1H-
NMR is consistent with structure; MS (FD) 492 (M+); Anal.
Calc' d for: G, 55.2:?; H, 6. 06; N, 14 . 86. Found: 55.33; H,
6.28; N, 13.24; IR (KBr) 3224, 3061, 3032, 2962, 2936, 2873,
1678, 1636, 1538, 1498, 1454, 1101 cm 1.
Preparation 49
H
O N NHBoc
O O
HN
N
N
N
Reaction of a compound of Preparation 47 (1.00 g, 580
mmol) , hexamethylene~imine (0.2 mL, 1 .7 mmol) , 1-
hydroxybenzotriazole hydrate (0.25 g, 1.9 mmol) and 1,3-
dicyclohexylcarbodii.mide (0. 4 g, 1 . 9 mmol ) in N, N-
dimethylformamide (50 mL) as described in Preparation 4 gave
0.76 g (68~) of the desired product as a tan foam: 1H-NMR is
consistent with structure; MS (FD) 660.2 (M+); Anal. Calc'd
for: C, 65.43; H, 7.32; N, 12.02. Found: C, 65.92; H, 7.86;
N, 11.71.
SUBSTITUTE SHEET (RULE 26)


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Example 12
H
~2
O O
HN
N
~,"~ N
2.67 HC1
Reaction of a compound of Preparation 49 (0.67 g, 1.0
mmol) and trifluoroacetic acid (4 mL, 52 mmol) in
dichloromethane (12 mL) for 1 h at ambient temperature,
followed by acidification with HC1 in ethyl acetate,
according to Preparation 4 gave 0.3 g (48$) of the desired
product as a white solid: 1H-NMR is consistent with
structure; MS (FD) 560.4 (M); Anal. Calc'd for: C, 58.77; H,
6.56; N, 12.01. Found: C, 56.48; H, 6.41; N, 12.06.
Preparation 50
NO 2
HN
Et0
O
To a solution of m-nitroaniline (1.0 g, 7.24 mmol)
stirred in anhydrous N,N-dimethylformamide (40 mL) at room
temperature was added a solution of a compound of
SUBSTITUTE SHEET (RULE 26)


CA 02340344 2001-02-12
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Preparation 5 (2.11 g, 8.69 mmol) in anhydrous N,N-


dimethylformamide
(10 mL). After
2.5 h, the reaction


mixture was diluted with H20 (70 mL) and extracted with


ethyl acetat e. Th~~ combined organic extracts were washed


with brine, dried (Na2S04), and concentrated to
i


g
ve a


yellow oil. Purification by radial chromatography (silica


gel, 10~-75~ ethyl acetate/hexanes) provided 1.65 g (76~)
of


the product (1:1 mixture of diastereomers) as an orange


solid. IH NMR
(300 MHz, CDC13)
d 7.47-7.53
(m, 3H), 7.33-


7. 41 (m, 7.20-7.25 (app. t, 1H, J = 8.1Hz) , 6. 81-6.
4H) , 85


(dd, 1H, J = B.OHz; 2.lHz), 5.10 (s, 1H), 4.12-4.26 (m, 2H),


1.20-1.25 (t, 3H, J = 7.lHz); 13C NMR (75.5 MHz, CDC13) d


271.0, 149.2, 196.5, 136.4, 229.6, 128.9, 128.5, 127.0,


119.1, II2.5, 107.2, 62.1, 60.3, 13.9; FD+ MS for C26H16N204


- 300; Anal. calcd. for C16H16N204 C, 63.99; H, 5.37; N,


9.33; Found: C, 64.77; H, 5.26; N, 9.17.


Preparation 51
~2N-.~N
')
N
Et0 \
To a slurry of sodium hydride (0.15 g of a 60$
dispersion in mineral oil, 3.86 mmol) stirring in N,N-
dimethylformamide (30 mL) at room temperature, was added a
solution of 6-nitrobenzimidazole (0.60 g, 3.68 mmol) in N,N-
dimethylformamide (10 mL). After 10 min, a solution of a-
bromophenylacetic acid ethylester in N,N-dimethylformamide
(10 mL) was added and the solution stirred for 4 h at room
temperature, quenched with water, and extracted with ethyl
SUBSTITUTE SHEET (RULE 26)


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acetate. The combined organic extracts were washed with
water, brine, dried (Na2S04) and concentrated. Purification
by silica gel chromatography (25$-75~ ethyl acetate/hexanes)
gave 0.580 g (5090) of the product (mixture of diastereomers)
as a yellow oil: 1H NMR (300 MHz, CDC13) d 8.72-8.73 (d,
1H, J = 2.lHz), 8.22-8.27 (dd, IH, J = 9.2Hz; 2.lHz), 8.15
(s, 1H), 7.44-7.50 (app. t, 3H, J = 6.9Hz), 7.34-7.41. (m,
3H), 6.19 (s, 1H), 4.26-4.39 (m, 2H), I.27-1.33 (t, 3H, J =
6.9Hz); FD+ MS for CI7H15N304 = 325; Anal. calcd. for
C17HI5N304: C, 62.'T6; H, 4 . 65; N, 12, 92; Found: C, 62. 89;
H, 4.92; N, 12.92.
Preparation 52
NO 2
HN
HO
i
To a solution o f a compound of Preparation 50 (0.81 g,
2.73 mmol) stirring .in dioxane (30 mL) at room temperature
was added LiOH~H20 (0.57 g, 13.6 mmol) and H20 (15 mL).
After 45 min, the mixture was concentrated to a volume of
approximately 20 mL.. The resulting aqueous solution was
diluted with H20 (7'i mL) and extracted with diethyl ether.
The aqueous layer was acidified with 1N HC1 and extracted
with ethyl acetate. The combined organic extracts were
washed with brine, dried (Na2S04) and concentrated to give
0.71 g (95$) of the product (1:1 mixture of diastereomers)
as a yellow solid: 1H NMR (300 MHz, CDC13) 8 7.48-7.55 (m,
SUBSTITUTE SHEET (RULE 2B)


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3H), 7.35-7.43 (m, 9H), 7.21-7.27 (app. t, 1H, J = 8.lHz),
6.81-6.85 (dd, 1H, J = 8.2Hz; 2.OHz), 5.16 (s, 1H); FD+ MS
for C14H12N2O4 = 272; Anal. calcd. for C14H12N204: C,
61.76; H, 4.44; N, 10.29; Found: C, 62.15; H, 4.52; N,
9.63.
Preparation 53
~ 2 N w/~.,. N
r
N
HO -.~ ~ /
O
to
To a solution of a compound of Preparation 51 (0.48 g,
1.48 mmol) stirring in dioxane (20 mL) at room temperature
was added LiOH~H20 f0.31 g, 7.38 mmol) and H20 (10 mL).
After 45 min, the reaction mixture was concentrated to a
volume of approximately 15 mL. The resulting aqueous
solution was dilutedi with H20 (75 mL) and extracted with
diethyl ether. The aqueous layer was acidified with 1N HCl
and extracted with ethyl acetate. The combined organic
extracts were washed with brine, dried (Na2S04) and
concentrated to give 0.450 g (>95~) of the product (1:1
mixture of diastereomers) as a light yellow solid: 1H NMR
(300 MHz, DMSO) b 8.63 (s, 1H), 8.56-8.57 (d, 1H, J =
2.lHz), 8.14-8.20 (dd, 1H, J = 9.2Hz; 2.lHz), 7.82-7.86 (d,
1H, J = 9.2Hz), 7.52-7.58 (m, 2H), 7.38-7.49 (m, 3H), 6.88
(s, 1H); FD+ MS for C15H11N304 = 297; Anal. calcd. for
C15H11N304: C, 60.61; H, 3.73; N, 14.14; Found: C, 59.59;
H, 9.16; N, 12.78.
SUBSTITUTE SHEET (RULE 26)


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Preparation 54
NO 2
HN
N
O --v O ~ i
y O
To a solution of a compound of Preparation 52 (0.75 g,
2.78 mmol), L-proline methyl ester hydrochloride (0.46 g,
2.78 mmol), 1-hydroxybenzotriazole hydrate (0.38 g, 2.78
mmol) and N,N-diisopropylethylamine (1.26 g, 9.72 mmol) in
anhydrous 1,2-dichlormethane (30 mL) stirring at room
temperature, was added 1-(3-dimethylaminopropyl)-3-
ethylcarbodiimide (0.585 g, 3.05 mmol). After 18 h, the
reaction mixture was diluted with H20 (50 mL) extracted with
ethyl acetate. The combined organic extracts were washed
with 10$ citric acid, sat'd aqueous sodium bicarbonate,
water, brine, dried (Na2S04) and concentrated. Purification
by radial chromatography (silica gel, 40~-75~ ethyl
acetate/hexanes) gave 0.56 g (53$) of the product (l:l
mixture of diastereomers) as a yellow solid: 1H NMR (300
MHz, CDC13) S 7.43-7.50 (m, 3H), 7.27-7.43 (m, 4H), 7.13-
7.20 (app, t, 1H, J = 7.5Hz), 6.83-6.91 (t, 1H, J = 5.8Hz),
5.14 (s, 1H), 4.52-4.58 (m, 0.5H), 4.41-4.47 (m, O.SH),
3.89-3.97 (m, 1H), 3.71 (s, 1.5H), 3.62 (s, 1.5H), 3.23-3.36
(m, 1H), 1.82-2,24 (m, 5H); 13C NMR (75.5 MH2, CDC13) d
172.2, 171.7, 168.7, 168.5, 149.0, 146.9, 146.5, 136.4,
135.9, 129.5, 129.4, 129.0, 128.8, 128.5, 128.2, 128.0,
SUBSTITUTE SHEET (RULE 26)


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127.8, 119.9, 119.6, 112.2, 112.0, 106.5, 106.5, 59.5, 59.4,
59.3, 59.3, 52.2, 52.0, 46.7, 46.7, 28.7, 28.6, 24.9, 24.5;
FD+ MS for C2pH21N305 = 383; Anal. calcd. for C2pH21N305:
C, 62.65; H, 5.52; N, 10.96; Found: C, 61.93; H, 5.62; N,
10.46.
Preparation 55
OZN y_ N
~ /
O
O~O
I
to
To a solution of a compound of Preparation 53 (0.43 g,
1.46 mmol), L-proline methyl ester hydrochloride (0.24 g,
1.46 mmol), 1-hydro:~cybenzotriazole hydrate (0.20 g, 1.46
mmol ) and N, N-diisopropylethylamine ( 0. 66 g, 5 . 10 mmol )
stirring in anhydrous 1,2-dichlormethane (30 mL) at room
temperature was added 1-(3-dimethylaminopropyl)-3-
ethylcarbodiimide (0.31 g, 1.60 mmol). After 18 h, the
reaction mixture wa~~ quenched with H20 (50 mL) and extracted
with ethyl acetate. The combined organic extracts were
washed with 10~ citric acid, saturated aqueous sodium
bicarbonate, H20, brine, dried (Na2S04) and concentrated.
Purification by radial chromatography (silica gel, 50~ ethyl
acetate/hexanes to 100$ ethyl acetate gradient) gave 0.25 g
(42$) of the a single diastereomer as a white foam solid:
1H NMR (300 MHz, CDC13) d 8.75-8.76 (b, 1H, J = 2.lHz),
8.28-8.32 (dd, 1H, J = 8.9Hz; 2.lHz), 7.91 (s, 1H), 7.45-
7. 58 (m, 6H) , 6.26 (s, 1H) , 4 . 65-4 .70 (m, 1H) , 3. 83-3 . 92 (m,
1H), 3.78 (s, 3H), 3.30-3.39 (m, 1H), 1.95-2.30 (m, 5H); FD+
SU8ST1TUTE SHEET (RULE 26)


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MS for C21H20N405 =~ 408; Anal. calcd. for C21H20N405: C,
61.76; H, 4.94; N, 13.72; Found: C, 61.24; H, 5.16; N,
13.10.
Preparation 56
~ H
''N ~ NHBoc
C, ~~
O NI-r
HN
N ~~ w
Ow-y' O
O
To a slurry of 5$ Pd/C (0.07 g) in ethanol (30 mL) was
added a solution of a compound of Preparation 54 (0.15 g,
0.39 mmol) in ethyl acetate (30 mL). The mixture was
treated with hydrogen gas (32 psi) at room temperature for 4
h on a Parr apparatus then carefully filtered through
celite. The resulting filtrate was evaporated to provide an
off-white solid foam which was dissolved in N,N-
dimethylformamide (30 mL). To this solution was added a
compound of Preparation 4 (0.16 g, 0.41 mmol), 1-
hydroxybenzotriazole hydrate (0.06 g, 0.41 mmol) and 1,3-
dicyclohexylcarbodiimide (0.09 g, 0.45 mmol). This solution
was stirred overnight at room temperature and subsequently
diluted with water (50 mL) then extracted with ethyl
acetate. The combined organic extracts were washed with
water, brine, dried (Na2S04) and evaporated to provide a tan
foam. Purification by radial chromatography (silica gel,
50$ ethyl acetate/he:~anes to 100$ ethyl acetate gradient)
yielded 0.23 g (82$) of the product (mixture of
diastereomers) as an off-white solid foam. 1H NMR (300 MHz,
CDC13) d 8.83-8.91 (rn, 1H), 7.44-7.51 (m, 2H), 7.20-7,36 (m,
SUBSTITUTE SHEET (RULE 26)


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8H), 6.88-7.06 (m, 3H), 6.32-6.38 (app. t, 1H, J = 6.9Hz),
5.28 (s, 1H), 5.12-5.19 (m, 1H), 9.88-4.91 (br. s, 1H),
4.48-4.60 (m, 3H), 4.17-4.24 (m, 1H), 3.64-3.72 (app. q, 2H,
J = 8.OHz), 3.62 (s, 3H), 3.39-3.52 (m, 1H), 3.28-3.39 (m,
1H) , 1 . 81-2.15 (m, 5H) , 1 .53-1. 57 (app. d, 3H, J = 7. 9Hz) ,
1.38 (s, 3H), 1.39 (s, 9H); FD+ MS for C39H4gN50g = 716;
Anal. calcd. for C3!aH4gN50g: C, 65.44; H, 6.90; N, 9.78;
Found: C, 65.23; H,, 7.43; N, 10.34.
Preparation 57
O ~ N~'NHBoc
O
O N
i
O ,..~ N
'' ''~1 N -'~
O O
To a slurry of 5$ Pd/C (0.042 g) in ethanol (30 mL) was
added a solution of a compound of Preparation 55 (0.08 g,
0.20 mmol) in ethyl acetate (30 mL). The mixture was
treated with hydrogen gas (32 psi) at room temperature for 4
h (Parr apparatus) then carefully filtered through celite.
The resulting filtrate was evaporated to provide a white
solid foam which was dissolved in N,N-dimethylformamide (20
mL). To this solution was added a compound of Preparation 4
(0.08 g, 0.20 mmol), i-hydroxybenzotriazole hydrate (0.03 g,
0.22 mmol) and I,3-d_Lcyclohexylcarbodiimide (0.05 g, 0.22
mmol). This solution was stirred overnight at room
temperature and subsE~quently diluted with water (50 mL) then
extracted with ethyl acetate. The combined organic extracts
were washed with watE:r and brine, dried (Na2S04) and
SUBSTITUTE SHEET (RULE 26)


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concentrated. Purification by radial chromatography (silica
gel, 75~ ethyl acetate/hexanes to 100$ ethyl acetate
gradient) yielded 0.10 g (66~) of the product (one
diastereomer) as an off-white solid foam: 1H NMR was
consistent with structure;
FD+ MS for C4pH48N60g = 740.
Example 13
H H \/
O N~~NH2
O NH'
i
HN
N
O
to O O
To a solution o:f a compound of Preparation 56 (0.17 g,
0.24 mmol) and aniso:Le (0.03 g, 0.26 mmol) stirring in
anhydrous dichloromethane (5 mL) at 0 °C was added
trifluoroacetic acid (1 mL). After 4 h, the reaction
mixture was quenched carefully with saturated aqueous sodium
bicarbonate extracted with ethyl acetate. The combined
organic extracts were: washed with sat'd aqueous sodium
bicarbonate, water, brine, dried (NaS204) and evaporated to
yield the desired product (1:I mixture of diastereomers) as
an off-white foam: 0.100 g (67~). 1H NMR was consistent
with structure; FD+ MS for C34H41N5~6 = 615; Anal. calcd for
C34H41N5~6~ C, 66.32; H, 6.71; N, 11.37; Found: C, 65.83;
H, 6.50; N, 6.50.
SUBSTITUTE SHEET (RULE 26)


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Example 14
w ~~ O~"N~'NH 3+CF 3CO0 '
I ~ ~... o
0 N
i
O ..~ N
N.
(~ 0
To a solution of a compound of Preparation 57 (0.080 g,
0.11 mmol) and anisole (0.0123 g, 0.114 mmol) stirring in
anhydrous dichloromeahane (5 mL) at 0 °C was added
trifluoroacetic acid (1 mL). After 4 h, the mixture was
quenched carefully with saturated sodium bicarbonate and
extracted with ethyl acetate. The combined organic extracts
were washed with saturated aqueous sodium bicarbonate,
water, brine, dried (NaS204) arid concentrated to yield foam
0.09 g (95$) of the desired product (one diastereomer) as an
off-white solid: 1H NMR was consistent with structure; FD+
MS for C35H4pN606-2CF3COOH = 640 (M-2CF3COOH); Anal. calcd.
for C39H42N6010F6: C, 53.92; H, 4.87; N, 9.67; Found: C,
51.86; H, 4.74; N, 9.54.
SUBSTITUTE SHEET (RULE 26)


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EXAMPLES PART 2A
Preparation la
~~ NHBoc
CO Me
2
tart-Butyloxycarbonyl-O-benzyl-D-serine methyl ester.
To a solution oi: t-butyloxycarbonyl-0-benzyl-D-serine
(25.0 g, 84.7 mmo7_) stirring in dimethylformamide (500
mL) at room temperature was added sodium bicarbonate
(14.2 g, 169 mmol) followed by methyl iodide (26.4 mL,
424 mmol). After 18 h, the reaction mixture was
concentrated to approximately 100 mL. Ethyl acetate was
added and the mixture washed with aqueous sodium
bicarbonate and brine. The organic extract was dried and
concentrated to give the desired compound (25 g, 96~) as
a light yellow oil: 1H NMR (300 MHz, CDC13) d 1.45 (s,
9H), 3.70 (m, 1H), 3.75 (s, 3H), 3.85 (m, 1H), 4.50 (m,
3H), 7.30 (m, 5H); MS (FD) m/e 310; Anal. calc'd for
C16H23N05: C, 62.12; H, 7.49; N, 4.53. Found: C, 62.31;
H, 7.49; N, 4.43.
Preparation lb
~2
CO Me
O-benzyl-D-serine methyl ester.
To a solution of tort-butyloxycarbonyl-O-benzyl-D-serine
methyl ester (BF8-Ei:O-275) (5.0 g, 16 mmol) stirring in
dichloromethane (40 mL) and anisole (1 mL) at 0 oC was
added trifluoroacetic acid (10 mL). After 4 h at room
temperature, a saturated aqueous solution of sodium
bicarbonate was added and the resulting mixture extracted
with ethyl acetate. The combined organic extracts were
SUBSTITUTE SHEET (RULE 26)


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washed with brine, dried over sodium sufate~ and
concentrated. The crude product was used in the next
step without further purification.
Preparation lc
H
'w O~' N
NHBoc
Me02C O
To a solution of 0-benzyl-D-serine methyl ester (the
product of Preparation lb) (65.9 mmol), boc-a-
aminoisobutyric acid (13.2 g, 65.4 mmol), 1-
hydroxybenzotriazole (8.8 g, 65.4 mmol), and N,N-
diisopropylethylami:ne (22.8 mL, 130.7 mmol) stirring in
dichloromethane (500 mL) at 0 °C was added 1-(3-
dimethylaminopropyl;l-3-ethylcarbodiimide (12.3 g, 71.9
mmol). After 18 h, ethyl acetate and ammonium chloride
(saturated aqueous ;solution) were added ands the resulting
mixture extracted with aqueous ammonium chloride, aqueous
sodium bicarbonate, and brine. The organic extracts were
dried over sodium sulfate and concentrated. Purification
by flash chromatography (25~ ethyl acetate/hexanes)
yielded the desired compound (21.6 g, 83$) as a white
solid: IH NMR (300 MHz, CDC13) d I.39 (s, 9H), 1.48 (s,
6H), 3.62 (dd, J = 3.4, 9.1 Hz, 1H), 3.70 (s, 3H), 3.85
(dd, J = 3. 4, 9.1 H;a, 1H) , 4. 48 (dd, J = 12.5, 22. 7 Hz,
2H), 4.75 (m, 1H), 4.92 (s, 1H), 7.11 (d, J - 8.6 Hz,
1H), 7.35 (m, 5H); MS (FD) m/e 395; Anal. calc'd for
C20H30N206: C, 60.90; H, 7.67; N, 7.10. Found: C,
61.02; H, 7.78; N, 7.10.
SUBSTITUTE SHEET (RULE 26)


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Preparation ld
H
O~N NHBoc
H02C O
To a solution of i~he product of Preparation lc (5.30 g,
13.4) stirring in dioxane (100 mL) /water (50 mL) at room
temperature was added lithium hydroxide (2.80 g, 67.3
mmol). After 18 h, water was added and the solution
concentrated. The resulting mixture was extracted with
diethyl ether. Brine was added to the aqueous layer and
the pH adjusted to 3.5 with 1 N HC1. The resulting
ZO mixture was extracted with ethyl acetate and the combined
organic extracts dried over sodium sulfate then
concentrated to yield the title compound (4.40 g, 86$) as
a white foam: 1H NMR (300 MHz, CDC13) d 1.39 (s, 9H),
1.45 (s, 3H), 1.47 (s, 3H), 3.68 (m, 1H), 3.95 (m, 1H),
4.54 (s, 2H), 4.70 (m, 1H), 5.51 (bs, 1H), 7.18 (d, J =
9.1 Hz, 1H), 7.25 (m, 5H), 9.90 (bs, 1H); MS (FD) m/e
381; Anal. calc'd for C1gH28N206: C, 59.99; H, 7.42; N,
7.36. Found: C, 59"74; H, 7.26; N, 7.30.
Preparation le
Et0 C N HAc
C/ CO Et
A solution of sodium ethoxide was generated by the
addition of sodium metal (52.89 grams, 2.3007 mol) over 3
hours to ethanol (1500 mL). To the sodium ethoxide
solution at ambient temperature was added a solution of
diethylacetamidomalonate (499.75 grams, 2.3007 mol)
dissolved in ethanol. (225 mL). The reaction mixture was
stirred for 1.5 hours at ambient temperature. 1-bromo-3-
phenylpropane (458.07 grams, 2.3007 mol) was added over
15 minutes and the reaction mixture was refluxed until
SUBSTITUTE SHEET (RULE 2S)


CA 02340344 2001-02-12
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complete as determined by hplc (16 hours). The reaction
mixture was concentrated to dryness and the residue
partitioned between ethyl acetate (1 x 1500 mL and 2 x
500 mL) and water (1500 mL). The ethyl acetate layers
were combined, washed with saturated sodium chloride
solution (9 x 500 mL), dried using sodium sulfate, and
concentrated to give 752.1 grams (98~) of the desired
compound as a light yellow solid. A 1.0 gram sample was
recrystallized from hexane:ethyl acetate (19:1, v:v) to
give a mp 84-86°C. 1H nmr ( CDC13): 8 1.18-1.23 (t,
6H), 1.37-1.50 (m, 2H), 2.02 (s, 3H), 2.34-2.41 (m, 2H),
2.58-2.62 (t, 2H), 4.16-4.24 (q, 4H), 6.76 (s, broad,
1H) , 7 .11-7.28 (m, 5H) . 13C nmr ( CDC13) : 8 13. 95,
23.03, 25.67, 31.85, 35.45, 62.46, 66.49, 125.40, 125.90,
128.27, 128.35, 14:1.77, 168.11, 168.94. MS (FIA ) m/z
336.3 ( [M+H]+) . IFt (KBr, cm 1) 1645.98 (amide) , 1744.76
(C=0) . Anal. Calcd.. for C18H25NO5: C, 64.46; H, 7.51; N,
4.17. Found: C, 64.60; H, 7.37; N, 4.39.
Preparation if
_ NHAc
/ C02H
(DL)-N-Acetyl-2-amino-5-phenylpentanoic Acid. A slurry
consisting of the product of Preparation le (249.15
grams, 0.7428 mol) and 2.5 N sodium hydroxide solution
was heated at 100°C for three hours. The reaction mixture
was cooled to 30°C and the pH adjusted to 5.0 using
concentrated hydrochloric acid. The solution was heated
to 100 °C and the pH was held at 5.0 using concentrated
hydrochloric acid as needed until the reaction was
complete as determined by hplc. The solution was
filtered while hot through diatomaceous earth. The
SUBSTITUTE SHEET (RULE 26)


CA 02340344 2001-02-12
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filtrate was cooled to 5-10 °C and the pH adjusted to 1.0
using concentrated hydrochloric acid. The resulting
slurry was stirred for 1 hour at 5 °C, filtered, and dried
in vacuum at 50 °C to give 160.34 grams (92$) of (DL)-N-
acetyl-2-amino-5-phenylpentanoic acid as a white powder,
mp 145-148 °C. 1H nmr ( DMSO-d6) : 8 1. 60-1 .71 (m, 4H) ,
1. 86 (s, 3H) , 2. 5E>-2.59 (m, 2H) , 4 . 19-4 .23 (m, 1H) , 7.16-
7.30 (m, 5H) , 8.14 (d, 1H) . 13C nmr ( DMSO-d6) : s 23.17
28.25, 31.55, 35.51, 52.55, 126.60, 129.14, 142.64,
170.25, 174.65. M:S ( FIA) m/z 236.2 (M+) . IR (KBr, cm
') 1609.17 (amidel, 1741.12 (C=0). Anal. Calcd. for
C13H1,N03: C, 66.36; H, 7.28; N, 5.95. Found: C, 66.41;
H. 7.15; N, 5.96.
Preparation la
NHAc
v
i s ~ CI 02H
(D)- N-Acetyl-2-amino-5-phenylpentanoic Acid. A solution
consisting of (DL)-N-acetyl-2-amino-5-phenylpentanoic
acid (438.0 grams, 1.862 mol), cobalt chloride (1.10
grams), 2N potassium hydroxide solution (931 mL, 1.862
mol) , and water (8000 mL) was adjusted to a pH of 8.0 by
the addition of 2N potassium hydroxide solution. To the
reaction mixture was added Acylase I (aspergillus
melleus, 39.42 gram~c) and vigorously stirred for 24 hours
at 40 °C while maintaining a pH of 8.0 by addition of 2N
potassium hydroxide. The resulting slurry was filtered.
The filtrate was adjusted to a pH of 2.0 giving a thick
slurry. The product was isolated by filtration, washed
with hexane (2000 mL) and dried in vacuum at 50 °C to give
188.52 grams (43~) of (D)-N-acetyl-2-amino-5-
phenylpentanoic acid. 1H nmr ( DMSO-d6): 8 1.59-1.74
SUBSTITUTE SHEET (RULE 26)


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WO 00/10565 PCTNS99l03525
-115-
(m, 4H), 1.86 (s, 3H), 2.57-2.60 (m, 2H), 4.22-4.26 (m,
1H) , 7 . 16-7 , 30 (m, 5H) , 8 . 02 (d, 1H) , 12 . 39 (s, broad,
1H) . '3C nmr ( DMSO-d6) : 8 23.18, 28.13, 31.66, 35.54,
52.58, 126.56, 129.10, 142.67, 170.12, 174.48. MS (FIA )
m/z 236. 1 (M+) . IR (KBr, cm-') 1625. 08 (amide) , 1700.24
(C=0) . Anal. Calcd. for C13H1,N03: C, 66.36; H, 7.28; N,
5.95. Found: C, 156.49; H, 7.00; N, 6.03.
Preparation lh
_ NH2
-i CO Et HCI
2
(D)-2-Amino-5-phenylpentanoic Acid, Ethyl Ester
Hydrochloride. A solution consisting of (D)-N-acetyl-2-
amino-5-phenylpentanoic acid (188.8 grams, 0.8024 mol),
ethanol (535 mL), and concentrated hydrochloric acid (268
mL, 3.21 mol) was warmed to 85 °C and monitored by hplc.
The reaction was determined to be incomplete by hplc at
14.5 hours and additional concentrated hydrochloric acid
(50 mL) was added. The reaction was determined to be
complete by hplc after 22.5 hours. Water was
azeotropically dist_~lled from the reaction by continuous
addition and distillation of 8000 mL of ethanol. The
ethanol was azeotropically distilled from the reaction by
the continuous addition and distillation of ethyl acetate
(2000 mL). Upon cooling the solution to 0 °C the product
crystallized. The solution containing the product was
stirred for 1 hour at 0 °C, filtered, and the cake dried
in vacuum at 40 °C to give 199.0 grams (96~) of 2-amino-5-
phenylpentanoic acid, ethyl ester hydrochloride, mp 117-
121 °C. 'H nmr ( DMSO-d6) : S 1.15-I .21 (t, 3H) , 1. 50-
1.89 (m, 4H), 2.48-2..67 (m, 2H), 3.92-3.98 (t, 1H), 4.08-
4.25 (m, 2H), 7.12-'7.29 (m, 5H), 8.76 (s, broad, 3H).
'3C nmr (DMSO-d6): b 13.90, 25.97, 29.52, 34.41, 51.71,
SUBSTETUTE SHEET (RULE 26)


CA 02340344 2001-02-12
WO 00/10565 PCT/US99/03525
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61.56, 124.91, 125.81, 128.24, 141.27, 169.35. MS (FIA )
m/z 222.3 (M') . 7:R (KBr, cm 1) 1741.14 (C=0) . [aj2o°
-11.17(c - 30.62 mg / 3mL, MeOH). Anal. Calcd. for
C~sH2oNO2Cl: C, 60.58; H, 7.82; N, 5.43. Found: C,
60.45; H, 7.67; N, 5.55.
Preparation li
H
N
NHBoc
O
Et0 O
A slurry consisting of N-t-BOC-a-aminoisobutyric acid
(90.64 grams, 0.446 mol), 2-chloro-4,6-dimethoxy-1,3,5-
triazine (75.90 grams, 0.425 mol), N-methyl morpholine
(88.13 grams, 0.871 mol), and diethyl ether (1000 mL) was
stirred at ambient temperature until complete as
determined by hpl.c (3 hours). The D-2-amino-5-
phenylpentanoic acid, ethyl ester hydrochloride (109.55
grams, 0.425 mol) was added and the reaction mixture
stirred for 16 hours at ambient temperature. The
reaction mixture was partitioned between 10$ citric acid
solution (1000 mL) and ethyl acetate (3 x 500 mL). The
organic phase was washed with 10$ citric acid solution (3
x 500 mL), saturated sodium bicarbonate solution (3 x 500
mL), water (1 x 500 mL), dried using sodium sulfate, and
concentrated to dryness. The residue was recrystallized
from hexane (3000 mL) to give 155.11 grams of the desired
compound: mp 97-99 °C. 'H nmr ( CDC13) : S 1.25-1.28 (t,
3H), 1.43 (s, 9H), 1.48 (s, 3H), 1.50 (s, 3H), 1.70-1.73
(m, 3H) , 1. 87-1 . 93 (:m, 1H) , 2 . 62-2 . 67 (m, 2H) , 4 . 16-4 . 21
(m, 2H), 4.57-4.62 (m, 1H), 4.95 (s, 1H), 6.96 (s, broad,
1H), 7.16-7.19 (m, 3H), 7.26-7.33 (m, 2H). 13C nmr
SUBSTITUTE SHEET (RULE 2B)


CA 02340344 2001-02-12
WO 00/10565 PCT/US99/03525
-117-
( CDC13) : $ 14.:13, 26.32, 27.17, 28.67, 32.47, 35.73,
52.54, 57.17, 6I.62, 126.21, 128.69, 128.79, 142.12,
154.99, 172.81, 174.69. MS (FIA ) m/z 407.5 ([M+H]+). IR
(KBr, cm 1) 1652.i'S, 1685.52 (amides), 1741.73 (C=O).
[oc]Z°o - 7.83 (c - 10.22 mg / lmL, MeOH) . W (0.1$
trifluoroacetic aced in water . acetonitrile) ~,",aX 215.6
nm. Anal. Calcd. for CZZH34N2O5: C, 65.00; H, 8.43; N,
6.89. Found: C, 65.23; H, 8.34; N, 6.94.
Preparation li
H
N NHBoc
I ~ HO- ' O O
A solution consisting of the product of Preparation li
(152.53 grams, 0.3752 mol) and tetrahydrofuran (884 mL)
was cooled to 5 °C. A solution consisting of lithium
hydroxide (26.96 grams, 1.126 mol) and water (1419 mL)
was added to the reaction dropwise over 10 minutes
maintaining a temperature of 5-10 °C. Ethanol (183 mL)
was added and the reaction stirred at 5-10 °C until
complete as determined by hplc (2 hours). The pH of the
reaction mixture was adjusted to 2.0 using 6 N
hydrochloric acid solution while maintaining 5-10 °C. The
product was extracted from solution with ethyl acetate (3
x 500 mL). The ethyl acetate extracts were combined,
dried using sodium sulfate, and concentrated to dryness
to give 141.51 grams (1000 of The desired compound: 1H
nmr ( DMSO-d6) : 8 1. 32-1. 37 (m, 15H) , 1. 57-I . 75 (m, 4H) ,
2.51-2.58 (m, 2H), 4.23-4.27 (m, 1H), 6.85 (s, broad,
1H), 7.15-7.28 (m, 5H), 7.42 (d, 1H), 12.5 (s, broad,
1H) . 13C nmr ( DMSO-d6) : 8 26.31, 27.85, 29.00,
31.86, 35.60, 52.53, 56.60, 78.95, 126.52, 129.05,
129.10, 142.69, 155.06, 174.40, 175.17. MS (FIA ) m/z
SUBSTITUTE SHEET (RULE 26)


CA 02340344 2001-02-12
WO 00/10565 PCT/ITS99/03525
-118-
379.5 ([M+H]+). Ilk (KBr, cm') 1641.98, 1692.22 (amides),
1719.72 (C=0). Ca]'°° _ -5.73 (c = 10.48 mg / lmL, MeOH}.
Anal. Calcd. for C.~;,H3°NZO5: C, 63.47; H, 7.99; N, 7.40.
Found: C, 63.25; H, 7.84; N, 7.46.
Preparation 1L
H
I N NH
J ~ ~
N O_ _ O O' _O
g OH
N-Methyl morpholine (4.79 mL, 2 eq, 47.3 mm) was added to
a stirred slurry of: N-Boc-a-aminoisobutyric acid (4.43 g,
2I . 7 mm, 1 eq) and 3. B9 g (21 . 7 mm, 1. 0 eq) of 2-chloro-
(4,6)-dimethoxy-1,3,5-triazine (CDMT) in 100 mL of
diethyl ether. After stirring the reaction mixture at
ambient temperaturE~ for 1.5 hours, D-tryptophan ester
hydrochloride was added. After stirring overnight, the
reaction mixture was quenched by the addition of 150 mL
of 10$ aqueous citric acid solution. The layers were
separated and the ether layer was washed with 50 mL of
saturated sodium bicarbonate solution and 50 mL of
water. Lithium hydroxide (2.43 g, 5 eq) was dissolved in
100 ml of water and the solution was added to the diethyl
ether solution and stirred vigorously for 4 hours at room
temperature. The layers were separated and the pH of the
aqueous layers was adjusted to 5.6 with 1M HC1. The pH
was then adjusted to 3.95 with 10$ citric acid solution
and the aqueous layer was extracted with 100 mL of ethyl
acetate. The ethyl acetate layers were washed with
brine, dried over magnesium sulfate and filtered. The
volatiles were removed under vacuum to give 82 $ yield of
the desired product as a white foam. 1H-NMR consistent
with structure.
SUBSTITUTE SHEET (RULE 26~


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Preparation 2A
o~
o /_
~''~ o
To a solution of 4-methoxyphenylacetic acid (98 g, 590
mmol) in absolute ethanol (300 mL) was added of p
toluenesulfonic acid (20 g, 105 mmol) . The reaction
mixture was heated to reflux and maintained at that
temperature for 5 lz then cooled to room temperature and
concentrated to dryness. The resulting oil was purified
by flash chromat=ography (silica gel, 20$ ethyl
acetate/hexanes) to give 102 g (89$) of the desired
product as a colorless oil: 'H-NMR (d, DMSO) 1.17 (t, J =
8 . 7 Hz, 3H) , 3. 56 (;;, 2H) , 3. 73 (s, 3H) , 4 . 05 (q, J = 7.2
Hz, 2H), 6.87 (d, J = 8.7 Hz, 2H), 7.17 (d, 8.7 Hz, 2H);
MS ( ion spray) 195. 3 (M+1 ) ; Anal . Calc' d for C1,H19O3: C,
68.02; H, 7.27. Found: C, 67.95, 7.17.
To a solution of the product of Preparation 2A (40 g, 200
mmol) in carbon tetrachloride (500 mL) was added N-
bromosuccinimide (37 g, 206 mmol) and hydrobromic acid
(4 drops of 48$ aqueous solution). The resulting mixture
was heated to reflux and maintained at that temperature
for 5 h then cooled to room temperature, filtered, and
concentrated. The :resulting oil was purified by flash
chromatography (silica gel, chloroform) to give 51.1 g
(94$) of the desired product as a colorless oil: 'H-NMR
SUBSTITUTE SHEET (RULE 2B)
Preparation 28


CA 02340344 2001-02-12
WO 00/10565 PC'T/US99/03525
-120-
(d, DMSO) 1.19 (t, J - 8.4 Hz, 3H), 3.77 (s, 3H), 4.18
(m, 2H) , 5. 88 (s, 1H) , 6. 95 (d, J = 8 . 4 Hz, 2H) , 7. 50 (d,
J = 8.4 Hz, 2H); MS (FD) 272, 274 (M+); Anal. Calc'd for
ClH~3Br03: C, 48.37; H, 4.80. Found: C, 48.52, 4.77.
Preparation 3
o-
o=N
N
~y0
O ~ ~
/ /
O
To a solution of the product of Preparation 2B (49.5 g,
181 mmol) stirring in dimethylformamide (500 mL) at room
temperature was added 4-nitroimidazole (20.5 g, 181 mmol)
and potassium carbonate (?5 g, 543 mmol). After 16 h,
the reaction was filtered and concentrated. The resulting
oil was partitioned between ethyl acetate and water and
extracted with ethyl acetate. The combined organic
extracts were washed with brine, dried over sodium
sulfate, filtered and concentrated. The resulting oil
was purified by flash chromatography (silica gel, 30-70~
ethyl acetates/hexanes gradient) to yield 33.6 g (6I~) of
the desired product as an orange oil that solidifies upon
standing: 'H-NMR (d, DMSO) 1.17 (t, J = 7.2 Hz, 3H), 3.78
(s, 3H) , 4.25 (q, .T = 7.2 Hz, 2H) , 6.57 (s, 1H) , 7.02 (d,
J = 8.7 Hz, 2H) , 7. 46 (d, J = 8.7 Hz, 2H) , 7. 92 (s, 1H) ,
8.38 (s, 1H) ; MS (ion spray) 306 (M+1) ; Anal. Calc' d for
C14H15N3~5: C, 55.08; H, 4.95; N, 13.76. Found: C, 54. 93;
H, 4.89; N, 13.82.
SUBSTITUTE SHEET (RULE 26~


CA 02340344 2001-02-12
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preparation 4
\. O N II ' N O / ,
O
O N
~N
-O \N~
O
To a slurry of 10$ palladium on carbon (6.0 g) in
tetrahydrofuran (30 mL) was added a slurry of the product
of Preparation 3 ;8.4 g, 27.5 mmol) in tetrahydrofuran
(30 mL). The reaction mixture placed under a hydrogen
atmosphere (40 mm Hg) using a Parr apparatus until the
reduction was complete then filtered through celite. To
the resulting solui:ion stirring at room temperature was
added the product of. Preparation ld (10.5 g, 27.5 mmol),
1-hydroxybenzotriazole (4.1 g, 30.3 mmol) and 1-(3-
dimethylaminopropyl)-3-ethylcarbodiimide (6.3 g, 30.3
mmol). After 16 h, the reaction mixture was concentrated
and the resulting oil was slurried in ethyl acetate and
filtered. The solution was diluted with water and then
extracted with ethyl acetate. The combined organic
extracts were washed with brine, dried over sodium
sulfate, filtered and concentrated. The resultant crude
material was purified by flash chromatography (silica
gel, 3~ methanol/chl.oroform) to give 14.4 g (83~) of the
desired product as a tan foam: 'H-NMR (d, DMSO) 1.78 (t, J
= 7 . 2 Hz, 3H) , 1 . 27-1. 32 (m, 15H) , 3 . 60 (m, 1H) , 3 . 67 (m,
1H) , 3. 76 (s, 3H) , 4 . 20 (d, J = 7 .2 Hz, 2H) , 4 . 44 (d, J =
3.0 Hz, 2H), 4.57 (m, 1H), 6.35 (s, 1H), 6.97 (d, J = 7.2
Hz, 2H), 7.20-7.35 (m, lOH), 7.40 (m, IH), 7.52 (s, 1H);
MS ( ion spray) 638 (M+1 ) ; Anal . Calc' d for C33H4sN5O8 : C,
62.15; H, 6.80; N, 10.98. Found: C, 62.41; H, 6.85; N,
11.09.
SUBSTITUTE SHEET (RULE 26)


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Preparation 5
\ O N N O
/~ / /~ O
O N O
C %N
O NJ
O
f
To a solution of the product of Preparation 4 (14.4 g, 23
mmol) stirring in dioxane (150 mL) at room temperature
was added a solution of of lithium hydroxide (0.65 g,
27.6 mmol) in water (75 mL) . After 20 min, the reaction
mixture was acidified to pH - 2.9 with 1 N hydrochloric
acid. To the resulting solution was added water and
ethyl acetate the mixture was extracted with ethyl
acetate. The combined organic extracts were washed with
brine, dried over. sodium sulfate and concentrated to
yield 13.0 g (93~) of the desired product as a yellow
foam: 'H NMR (d, DMSO) 1.25-1.40 (m, 15H), 3.65-3.70 (m,
2H), 3.76 (s, 3H), 4.44 (d, J - 3.4 Hz, 2H), 4.57 (m,
1H), 6.20 (s, 1H), 6.97 (d, J - 3.4 Hz, 2H), 7.15-'1.35
(m, lOH), 7.42 (m, 1H), 7.53 (s, 1H), 10.2 (s, 1H); MS
(ion spray) 610. 7 (M+1) ; Anal . Calc' d for ~31H3gN5Og: C,
61.07; H, 6.45; H, 11.49. Found: C, 60.90; H, 6.43; N,
11.32.
SUBSTITUTE SHEET (RULE 2S)


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Preparation 6
O N~N O
O~ ~
O O
/ '" N
J
N
O
O
To a solution of the product of Preparation 5 (g.p
8,13.0 mmol) stirring in dimethylformamide (150 mL) at
room temperature was added 4-methylpiperidine (1.6 mL,
13.0 mmol), 1-hydroxybenzotriazole (2.0 g, 14.3 mmol) and
1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (3.0 g,
14.3 mmol). After 16h, the reaction mixture was filtered
and concentrated. The resulting material was partitioned
between ethyl acetate and water and extracted with ethyl
acetate. The combined organic extracts were washed with
brine, dried ove r sodium sulfate, filtered and
concentrated to dryness. The resulting crude material
was purified by fla;;h chromatography (silica gel, 3$
methanol/ chloroform) to yield 7.65 g (85~) of the
desired product as a yellow foam: 'H-NMR (d, DMSO) 0.2
(m, 1H), 0.50 (d, ~T = 6.0 Hz, 1.5 H), 0.80 (d, J = 6.0
Hz, 1.5 H), 1.05 (m, 1H), 1.22-1.45 (m, 15H), 1.50-1.65
(m, 4H), 2.65 (m, lH), 3.00 (m, 1H), 3.55 (m, 1H), 3.65
(m, 1H), 3.75 (s, 3H), 4.37 (m, 1H), 4.40-4.50 (m, 2H),
4 . 60 (m, 1H) , 6. 62 (d, J = 13 Hz, 1H) , 6. 98 (t, J = g, 4
Hz, 2H), 7.10-7.45 (m, 11H), 10.15 (br s, 1H); MS (ion
spray) 691 . 3 (M+1 ) ; Anal . Calc' d for C37H50N6O~ ~ 0 . 6H20: C,
63.34; H, 7.35; N, 11.98. Found: C, 63.25; H, 7.03;
11.87.
SUBSTITUTE SHEET (RULE 26)


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WO 00/10565 PCTNS99/03525
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Examples 1 and 2
W. N N
O
i' ~~ O
O N
~N 2HC1
~N N.J
/ ' ~ v
0
0
To a solution of t:he product of Preparation 6 (7.26 g,
10.5 mmol) stirring in dichloromethane (25 mL) at room
temperature was added trifluoroacetic acid (10 mL).
After 4 h, the reaction mixture was poured into a
saturated solution of sodium bicarbonate extracted with
chloroform. The combined organic extracts were washed
with brine, dried over sodium sulfate, filtered and
concentrated to yield 6. 12 g ( 99$ ) of the free base as a
tan foam. The diastereomeric material (3.0 g) was
chromatographed on an 8 x 15 cm Prochrom column packed
with Kromasil CHI-~DMP chiral phase using an eluent
mixture of 3A alcohol (13$ by v), dimethylethylamine
(0.2~ by v) in heptane at a flow rate of 250 mL/min to
provide the individual diastereomers in pure form:
Example 1. Isomer 1 To a solution of the purified isomer
in ethyl acetate was added a saturated solution of
hydrochloric acid in diethyl ether. The resulting slurry
was concentrated to dryness to yield 1.1 g (37$) of the
desired product as a white solid: 'H NMR (d, DMSO) 0.50
(d, J = 6. 0 Hz, 1 . 5 Fi) , 0. 80 (d, J = 6. 0 Hz, 1. 5 H) , 1 .16
(m, 1H), 1.35 (m, 1Fi), 1.50-1.70 (m, 8H), 2.60-2.70 (m,
2H), 3.03 (m, 1H), 3.65-3.80 (m, 6H), 4.40 (m, 1H), 4.53
(s, 2H), 4.75 (m, 1H), 6.90-7.08 (m, 3H), 7.25-7.45 (m,
9H) , 8. 20-8 . 40 (m, 41:i) , 8 . 61 (d, J = 7. 5 Hz, 1H) , 11.15
(br s, 1H); tR = 7.93 min; MS (ion spray) 591.6 (M+1);
SUBSTITUTE SHEET (RULE 26)


CA 02340344 2001-02-12
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Anal . Calc' d for C;ZH42N6O5'2HC1: C, 57. 92; H, 6. 69; N,
12.66. Found: C, 57.72; H, 6.47; N, 12.42.
Example 2. Isomer 2 To a solution of the purified isomer
in ethyl acetate was added a saturated solution of
hydrochloric acid in diethyl ether. The resulting slurry
was concentrated to yield 0.98 g (33$) of the desired
product as a white solid: 'H NMR (d, DMSO) 0.50 (d, J =
6.0 Hz, 1.5 H), 0.80 (d, J - 6.0 Hz, 1.5 H), 1.16 (m,
1H), 1.35 (m, IH), 1..50-1.70 (m, 8H), 2.60-2.70 (m, 2H),
3.03 (m, 1H), 3.65--3.80 (m, 6H), 4.40 (m, 1H), 4.53 (s,
2H), 4.75 (m, 1H), 0.90-7.08 (m, 3H), 7.25-7.45 (m, 9H),
8.20-8. 40 (m, 4H) , 8. 61 (d, J = 7. 5 Hz, 1H) , lI .15 (br s,
1H); tR - 11.78 min; MS (ion spray) 591.6 (M+1); Anal.
Calc' d for C32H42N6O5'2 .2HC1: C, 57.29; H, 6. 64; N, 12.53.
Found: C, 57.23; H, 6.29; N, 12.57.
Preparation 7
N~N O
o O
~~N
N N.J
O O ~ \
O
Reaction of the product of Preparation 5 (0.6 g, I.p
mmol), 4-(4-fluorobenzoyl)piperidine hydrochloride (0.24
g. 1.0 mmol), triet:hylamine (0.15 mL, 1.1 mmol), 1-
hydroxybenzotriazole (0.16 g, 1.1 mmol), and I-(3-
dimethylaminopropyl)-:3-ethylcarbodiimide (0.23 g, 1.1
mmol) in dimethylformamide (40 mL) as described in
Preparation 6 gave 0..58 g (73$) of the desired product as
SUBSTITUTE SHEET (RULE 2B)


CA 02340344 2001-02-12
WO 00/t 0565 PCT/US99/03525
-12 6-
a tan foam: 'H-NMR (d, DMSO) 1.20-1.90 (m, 18H), 1.40-
1.90 (m, 3H), 2.83 (m, 1H), 3.55-3.73 (m, 3H), 3.75 (s,
3H), 3.85 (m, 1H),. 4.45 (d, J - 3.8 Hz, 2H), 4.60 (m,
1H), 6.65 (d, J = :L0.93 Hz, 1H), 6.95-7.05 (m, 2H), 7.10-
7.20 (m, 2H), 7.20-7.50 (m, 11H), 8.00-8.10 (m, 2H),
10.15 (br s, 1H); MS (FD) 798.7 (M+); Anal. Caic'd for
C43H51FN60g: C, 64.65; H, 6.43; N, 10.53. Found: C, 64.38;
H, 6.48; N, 10.61.
Examples 3 and 4
\ N~N
\O
O N
F / ~N
\I N N
O O ~ \
O
Reaction of the product of Preparation 7 (0.53 g, 0.66
mmol) and trifluoroacetic acid (2 mL) in dichloromethane
(6 mL) as described i.n Example 1 gave 0.34 g (74$) of the
desired mixture of diastereomers as a tan foam. This
material (0.11 g) was purified by HPLC (8 x 15 cm
Prochrom column packed with Kromasil'~ CHI-DMP chiral
phase, eluent mixture of 3A alcohol and
dimethylethylamine in heptane) to provide the individual
diastereomers which were converted to their repsective
hydrochloride salts as desribed in Example 1.
Example 3. Isomer 1. 1H-NMR (d, DMSO) 1.15-1.20 (m, 6H),
1.20-1.60 (m, 3H), 1.70 (m, 1H), 2.90 (m, 1H), 3.55-3.70
(m, 4H), 3.75 (s, 3H), 3.85 (m, 1H), 4.40 (m, 1H), 4.40-
4.55 (m, 2H), 4.60 (m, 1H), 6.65 (d, J - 11 Hz, 1H),
7.00-7.05 (m 2H), 7.20 (m, 1H), 7.20-7.40 (m, 13H), 8.00-
8.10 (m, 2H), 20.40 (br s, 1H); tR - 6.4 min; MS (ion
SUBSTITUTE SHEET (RULE 26)


CA 02340344 2001-02-12
WO 00/10565 PCT/US99/03525
-127-
spray) 699. 7 (M+1 ) ; Anal . Calc' d for C38Hq3FN~O~: C, 65. 31;
H, 6.20; N, 12.03. Found: C, 65.08; H, 6.18; N, 11.87.
Example 4. Isomer 2 'H-NMR (d, DMSO) 1.15-1.20 (m, 6H),
1.20-1.60 (m, 3H), 1.70 (m, 1H), 2.90 (m, 1H), 3.55-3.70
(m, 4H), 3.75 (s, 3H), 3.85 (m, 1H), 4.40 (m, 1H), 4.40-
4.55 (m, 2H), 4.60 (m, 1H), 6.65 (d, J - 11 Hz, 1H),
7.00-7.05 (m 2H), i'.20 (m, 1H), 7.20-7.40 (m, 13H), 8.00-
8.10 (m, 2H), 10.40 (br s, 1H); tR - 8.0 min; MS (high
res) calc'd fox C3gH~gqFN60s: 699.3306. Found: 699.3313.
Preparation 8
O N~N O
O
O N
N
O
Reaction of the product of Preparation 5 (1.0 g, 1.7
mmol), piperidine (0.17 mL, 1.7 mmol), 1-
hydroxybenzotriazole (0.25 g, 1.9 mmol), 1-(3-
dimethylaminopropyl)-3-ethylcarbodiimide (0.4 g, l.g
mmol), and dimethy:Lformamide (15 mL) as desribed in
Preparation 6 gave 0.7 g (60~) of the desired product as
a tan foam: 'H-NMR (d, DMSO) 0.97 (m, 1H), 1.25-1.40 (m,
15H), 1.40-1.55 (m, 7H), 3.30-3.45 (m, 2H), 3.60 (m,
1H), 3.67 (m, 1H), 3.75 (s, 3H), 4.45 (d, J - 3.4 Hz,
2H), 4.57 (m, 1H), 6.62 (s, 1H), 6.98 (d, J - 8.7 Hz,
2H), 7.13 (m, 1H), 7.25-7.45 (m, lOH), 10.15 (br s, 1H);
MS (ion spray) 677. 5 (M+1 ) ; Anal . Calc' d for C3sH48N60,: C,
63.89; H, 7.15; N, 12.42. Found: C, 63.97; H, 6.99; N,
12.44.
SUBSTITUTE SHEET (RULE 26)


CA 02340344 2001-02-12
WO 00/10565 PCT/US99/035Z5
-128-
Example 5
\r O N N
/ O
O N
C %N 2HC1
N
i ~ \
O
O
Reaction of the product of Preparation 8 (0.68 g, 1.0
mmol) and trifluoroacetic acid (2 mL) in dichloromethane
(6 mL) as described. in Example I gave 0.6 g (93g) of the
desired product as a white solid: 'H-NMR (d, DMSO) 0.95
(m, 1H), 1.30-I.60 (m, 11H), 3.20-3.40 (m, 3H), 3.60-3.75
(m, 3H), 3.78 (s, :3H), 4.50-4.55 (m, 2H), 4.75 (m, 1H),
6.80 (s, 1H), 7.05 (d, J - 9.0 Hz, 2H), 7.25-7.35 (m,
7H), 7.37 (d, J - 8.7 Hz, 2H), 8.10 (m, 1H), 8.20-8.30
(m, 3H), 8.58 (d, J - 7.6 Hz, 1H), 11.00 (br s, 1H); MS
(ion spray) 577. 4 (M+1 ) ; Anal . Calc' d for C3IHqON6O5~2 .2HC1:
C, 56.68; H, 6.48; N, 12.79. Found: C, 56.70; H, 6.64;
N, 12.37.
Preparation 9
N N O
O O
''O
/ ,."' N
J
N
N
/ O O
O
Reaction of the product of Preparation 5 (1.42 g, 2.3
mmol), d-proline methyl ester (0.3 g, 2.3 mmol), 1-
SUBSTITUTE SHEET (RULE 26)


CA 02340344 2001-02-12
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-129-
hydroxybenzotriazole (0.35 g, 2.5
mmol), and 1-(3-
dimethylaminopropyl)-3-ethylcarbodiimide
(0.53 g, 2.5
mmol) in tetrahydrofuran (15 mL)
as described in
Preparation 6 gave 0.99 g (60$) of
the desired product as
a white foam: 1H-NMR (d, DMSO)
1.25-1.40 (m, 18H), 1.75-


1.90 (m, 2H), 2.40 (m, 1H), 3.3 0 (m, 1H), 3.60-3.80 (m,
7H), 4.40 (m, 1H), 4.45-4.50 (m, 2H), 4.57 (m, 1H), 6.50


(m, 1H), 6.95-7.05 (m, 2H), 7.10-7.40 (m, 11H), 10.20
(br


s, 1H); MS (ion spray) 721.3 (M+1);Anal. Calc'd for
lO C3~Hq8N6Og: C, 61.65,; H, 6.71; 11.66.Found: C, 61.42;
N,


H, 6.43; N, 11.65.


Example 6
O N N
/ '~ O
O N
~N
J
N 2HC1
O O O
O
Reaction of the product of preparation 9 (0.87 g, 1.2


mmol) and trifluoroacetic acid (2 mL) in dichloromethane


(6 mL) as described in Example 1 gave 0.58g (70$) of the


desired product: I.40-1.60 (m, 6H),
'H-NMR (d,
DMSO)


1.75-1.95 (m, 3H), 2.20 (m, 1H), 2.95 (m, 1H), 3.60-3.80


(m, 9H), 4.40 (m, 7_H), 4.50-4.55 2H),4.75 (m, IH),
(m,


6.70 (s, 1H), 7.00 (t, J - 8.7 Hz, 2H),7.40-7.45 (m,


SUBSTITUTE SHEET (RULE 26)


CA 02340344 2001-02-12
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9H), 8.05 (m, 1H),, 8.20-8.30 (m, 3H), 8.55 (m, 1H), 10.95
(m, 1H); MS (ion spray) 621.5 (M+1); Anal. Calc'd for
C32HqpNgO~~2.3HC1: C, 54.55; H, 6.05; N, 11 . 93. Found: C,
54.96; H, 5.81; N, 11.79.
Preparation 10
N
-NFIH o c
O O
N
~O
O/
To a suspension of 5$ palladium on carbon (1.75 g) and
tetrahydrofuran (120 mL) was added the product of
Preparation 3 (3.51 g, 11.5 mmol). The reaction mixture
was placed under a hydrogen atmosphere (40 mm Hg) on a
Parr apparatus for 2 h then filtered through celite. The
filtrate was subsequently added to a solution of the
product of Preparation lj (4.33 g, 11.5 mmol), 1-(3-
dimethylaminopropyl)-3-ethylcarbodiimide (2.60 g,
12.6mmo1) and 1-hyd:roxybenzotriazole (1.72 g, 12.6 mmol)
stirring in tetrahyclrofuran (50 mL) at 0°C. After 16 h at
room temperature, the reaction mixture was concentrated.
The resulting residue was dissolved in ethyl acetate,
filtered and the resulting filtrate concentrated. The
crude residue was purified by flash chromatography
(silica gel, 90 ~; ethyl acete/hexanes to 10 $
methanol/ethyl acetate gradient ) to give 4 . 5 g ( 62 $ ) the
desired product as a light orange foam: 'H NMR consistent
with structure; MS (IS) m/e 636 (M + 1). Anal.
(C39Hq5N5O-,) C, H, N.
SUBSTITUTE SHEET (RULE 26)


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Preparation 11
N
NHBoc
O O
N
N
HO
O
O
To a solution of the product of Preparation 10 (1.01 g,
1.59 mmol) stirring in tetrahydrofuran (30 mL) and water
(15 mL) at room temperature was added lithium hydroxide
(0.26 g, 6.30 mmol). After 25 min, the reaction mixture
was concentrated and the resulting residue was diluted
with water and extracted with diethyl ether. The aqueous
extracts were acid~_fied to pH 2-3 with 1N hydrochloric
acid and then extracted with ethyl acetate. The combined
organic extracts were washed with brine, dried with
sodium sulfate and concentrated to provide 0.96 g (99 g)
of the desired compound as a light tan foam that was used
without further purification: 1H NMR consistent with
structure; MS (IS) m/e 608 (M + 1) . Anal. (C32H91NSO~) C:
calcd, 63.25; found, 62.68, H, N.
Preparation 12
N
NHBoc
O O
N
1 N
N
O
O
SUBSTITUTE SHEET (RULE 26)


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To a solution of the product of Preparation 11 (0.93 g,
1.53 mmol) stirring in dichloromethane (25 mL) at room
temperature was added N-methylmorpholine (0.20 mL, 1.83
mmol) and of 2-chloro- (4, 6) -dimethoxy-l, 3, 5-triazine
(0.35 g, 1.99 mmol). After 1 h, 4-methylpiperidine (0.20
mL, 1.68 mmol) was added and the resulting mixture was
stirred room temperature for 2 h at which time 2-chloro-
(4, 6) -dimethoxy-1, ?., 5-triazine (0.10 g, 0.70 mmol) was
added. After 1 h, the reaction mixture was concentrated
and the resulting residue purified by flash
chromatography (silica gel, ethyl acetate/methanol
gradient) to give t=he desired compound as a light yellow
solid foam (0.875 g, 83$); 'H NMR consistent with
structure; MS (IS) :m/e 689 (M + 1) .Anal. (C3eH52N6O6) C,H,N.
Example 7
I ~ , N
NH2
O O
N ~2HC1
1 N
N
I
O / /
O
To a solution of the product of Preparation 12 (0.77 g,
1.12 mmol) and anisole (0.13 mL, 1.13 mmol) stirring in
dichloromethane (20 mL) at 0 C was added trifluoroacetic
acid. After 3-4 h, the reaction mixture was warmed to
room temperature and then quenched by pouring over cold
saturated aqueous sodium bicarbonate. The organic layer
was collected and the aqueous layer was extracted twice
with dichloromethane. The combined organic extracts were
washed with aqueous sodium bicarbonate, water, brine,
SUBSTITUTE SHEET (RULE 26~


CA 02340344 2001-02-12
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then dried over sodium sulfate and concentrated. The
resulting material was purified by flash chromatography
(silica gel, 5$ methanol/ 95~ ethyl acetate gradient to
5~ triethylamine/T.O~ methanol/ 85a ethyl acetate) to
provide 0.63 g (95 ~) of the desired mixture of
diastereomers as an off-white solid foam. The mixture
(190 mg) was resolved by,chiral HPLC [Kromasil packing
material, 15$ 3A alcohol/ 85$ heptane (w/ 0.2$
dimethylamine)] to provide the two desired diastereomers.
To a solution of di.astereomer 2 (65 mg) (retention time =
9.00 min) stirrings in ethyl acetate (5 mL) was added
saturated solution of hydrochloric acid in diethyl ether.
The resulting whitf~ precipitate was collected by vacuum
filtration and rinsed with diethyl ether to provide the
desired compound (60 mg) as a white amorphous solid: 1H
NMR consistent with structure; MS (IS) m/e 589 (M + 1),
Anal. (C33HqqN6Oq~2HC1) C, H, N.
Preparation 13
O
N
CH~O
To a solution of Preparation 3 (3.00 g, 9.84 mmol)
stirring in tetrahydrofuran (10 mL) and ethanol (5 mL)
was added to sodium hydroxide (20 mL of a 5 N aqueous
solution). The resulting smixture was stirred at ambient
temperature until hydrolysis was complete and
subsequently acidic=ied to pH 2.0 with aqueous
hydrochloric acid. The reaction mixture was extracted
with ethyl acetate, dried over sodium sulfate, and
SUBSTITUTE SHEET (RULE 26)


CA 02340344 2001-02-12
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concentrated. The resulting carboxylic acid was combined
with pyrrolidine (0.710 g,10 mmol),
1-
hydroxybenzotriazole hydrate (1.35 g, 10 mmol) and 1,3-
dicyclohexylcarbodiimide (2.06 g, 10.0 mmol) stirring in
tetrahydrofuran (100 mL) at room temperature. After 18
h, the mixture was. concentrated, the residue slurried in
ethyl acetate then filtered and concentrated.
Purification by flash chromatography (silica gel,
chloroform/methanol.) provided afford 2.74 g (84$) of the
desired product: MS: (M+H)' 331.2; 'H NMR (300 MHz,
DMSO-d6) b 8. I9 (d, 1H, J = 1.51 Hz) , 7. 80 (d, 1H, J = 1.51
Hz) , 7. 45 (d, 2H, ~J = 8. 67 Hz) , 7. 02 (d, 2H, J = 8. 67 Hz) ,
6.58 (s, 1H), 3.7T (s, 3H),3.75-3.60 (m, 1H) 3.45-3.30
(m, 2H) , 2. 90-2. 75 (m, 1H} 1. 95-1. 60 (m, 4H) ; Anal . Calcd.
for C,6H18N404: C, 58.18; H, 5.49; N, 16.96. Found: C,
58.44; H, 5.45; N, 16.87.
The product of Preparation 13 (1.13 g, 3.42 mmol) was
added to a mixture of 10$ palladium/carbon (0.65 g) and
palladium/black (0.1:5 g) in tetrahydrofuran (40 mL) and
the mixture shaken under hydrogen (38 psi) in a Parr
apparatus. After reduction was complete, the reaction
mixture was filtrated through celite and the filtrate
immediately combined: with 1,3-dicyclohexylcarbodiimide
(0.71 g, 3.45mmo1), :1-hydroxybenzotriazole (0.46 g, 3.40
SUBSTITUTE SHEET (RULE 26)
Preparation 14


CA 02340344 2001-02-12
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-135-
mmol), the product: of Preparation lj (1.30 g, 3.44 mmol)
and additional tetrahydrofuran (60 mL). After stirring
overnight at amlbient temperature, the mixture was
concentrated and the residue slurried in ethyl acetate
then filtered. The filtrate was concentrated and the
residue purified by flash chromatography(silica gel,
chloroform/methanol) which afforded I.50g (66~) of the
desired product which was used without further
purification.
To a solution of the product of Preparation 14 (1.45 g,
2.20 mmol) in dichloromethane (30 mL) was added
triflouroacetic acid (10 mL). After 2 h, the mixture was
concentrated and thE~ residue treated with excess aqueous
sodium bicarbonate .and extracted. The combined organic
extracts were concentrated and the resulting residue was
purified by flash chromatography (silica gel,
chloroform/methanol) to provide 0.68 g of the desired
product as a yellow solid: MS: (M+H)' 561.3. 'H NMR was
consistent with product. Anal. Calcd. for C3lHqpN6Oq~0.2
CHC13: C, 64.11; H, 6.93;N, 14.38. Found: C, 64.19; H,
7.19; N, 14.50. The isomeric mixture (1.72 g) was
separated as previously described in Example 7 to provide
0.64 g of isomer I (t,~ = 7.50 min) and 0.49 g of isomer 2
(tR - 10.15 min) . Isomer 2 (486 mg, 0.87 mmol) was
SUBSTITUTE SHEET (RULE 26)
Example 8


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dissolved in a minimal amount of ehtyl acetate and
treated with an excess of saturated hydrochloric acid in
ethyl acetate. Concentration and subsequent evaporation
from diethyl ether allowed for recovery of 580 mg of an
off-white solid: NfS: (M+H)' 561.3, 562.4. 1H NMR was
consistent with product. Anal. Calcd. for C31H40N6Oq~3.0
HC1: C, 55.57; H, 6.47; N, 12.54. Found: C, 56.40; H,
6.43; N, 12.20.
Preparation 15
..3.
The product of Preparation 13 (0.85 g, 2.57 mmol) was
combined with 10~'s palladium/carbon (0.50 g) and
palladium/black ( 0. J.5 g) in tetrahydrofuran ( 40 mL) and
the mixture shaken under a hydrogen atmosphere (38 psi)
in a Parr apparatus.. After reduction was complete, the
catalyst was removed by filtration through celite and the
amine/tetrahydrofuran solution was immediately combined
with 1,3-dicyclohexy.lcarbodiimide (0.53 g, 2.57mmo1), 1-
hydroxybenzotriazole (0.35 g, 2.57 mmol), the product of
Preparation 1L (1.00 g, 2.57 mmol) and additional
tetrahydrofuran (60 mL). After stirring overnight at
ambient temperature, the mixture was concentrated and the
residue slurried in ethyl acetate and filtered. The
filtrate was concentrated and the residue purified by
flash chromatography(silica gel, chloroform/methanol)
SUBSTITUTE SHEET (RULE 26)


CA 02340344 2001-02-12
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which gave 1.62 g of the desired product which was used
without further purification.
The compound of Preparation 15 (1.57 g, 2.34 mmol) was
dissolved in dichloromethane (25 mL) and triflouroacetic
acid (10 mL) added. The resulting mixture was stirred at
ambient temperature for 2.5 h, concentrated, and the
residue treated with excess aqueous sodium bicarbonate.
The aqueous mixture was extracted with ethyl acetate and
the combined organic: extracts concentrated and dried. The
residue was chromatographed over silica gel
(chloroform/methanol) to provide 0.71 g (53 s) of the
desired product: MS: (M+H)+ 572.5. 'H NMR was consistent
with product. Anal. Calcd. for C31H3,N~04~0.35 CHC13: C,
61.38; H, 6.19; N, 15.98. Found: C, 61.36; H, 6.11; N,
16.08. The isomeric mixture (2.16 g) was separated as
previously described in Example 7 to provide 1.10 g of
isomer 1 (tR - 10.34 min) and 0.80 g of isomer 2 (tR -
13.70 min). The product derived from isomer 2 (0.80 g,
1.40 mmol) was dissolved in a minimal amount of ethyl
acetate arid the resu:Lting solution treated with an excess
of hydrochloric acid in ethyl acetate. The solution was
then concentrated to provide 0.88 g (82 ~) of the desired
product as an off white solid: MS: (M+H)+ 572.3, 573.4.
SUBSTITUTE SHEET (RULE 26)
Example 9


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'H NMR was consistent with product. Anal. Calcd. For
C3,H3,N-,04'3.0 HC1: C, 54.67; H, 5.92; N, 14.40. Found: C,
54.25; H, 5.89; N, 13.35.
Preparation 16
O-
O'Nt
N
OH
CHzO
To a solution of t:he product of Preparation 3 -411159-
(5.75 8,18.9 mmol) stirring at room temperature in
tetrahydrofuran (10 mL) was added sodium hydroxide (25 mL
of a 5 N aqueous solution) along with water (15 mL) and
ethanol (10 mL). After hydrolysis was complete, the
mixture was acidified to pH 2.0 with aqueous hydrochloric
acid and extracted. The combined organic extracts were
dried, filtered, and concentrated to give the desired
product in quantitative yield as a tan solid: 1H NMR (300
MHz, DMSO-ds) b 14. 05~-13. 60 (bs, 1H) , 8. 34 (s, 1H) 7. 90 (s,
1H), 7.45 (d, 2H, ~T - 8.67 Hz), 7.00 (d, 2H, ,T - 8.67
Hz), 6.42 (s, 1H), 3.77 (s, 3H). FDMS: 277 (M)+ Anal.
Calcd. for ClpHI1N3O5~0.67 H20: C, 49.82; H, 4.30;N, 14.52.
Found: C, 50.05; H, 4.01; N, 14.12.
;SUBSTITUTE SHEET (RULE 26)


CA 02340344 2001-02-12
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Preparation 17
p_
o-N.
N
\ w
CH~O
The compound of Preparation 16 (2.50 g, 9, 0 mmol) was
combined with ac(ueous dimethylamine(40$,1.15 mL,9.0
mmol), 1-hydroxy-benzotriazole hydrate(1.22 g, 9_0
mmol)and 1,3-dicyclohexylcarbodiimide (1.86 g, 9.0 mmol)
in tetrahydrofuran (60 mL) and the mixture stirred at
ambient temperatures. After 18 h, the mixture was
concentrated and the residue slurried in ethyl acetate
and filtered. The filtrate was concentrated and the
resulting residue purified by flash chromatography
(silica gel, chloro:form/methanol) to afford 1.83 g
of the desired product : 1H NMR ( 300 MHz, DMSO-d6) 8 8 .14
(s, 1H) 7.76 (s, 1H) , 7.42 (d, 2H, J = 8. 67 Hz) , 7.00 (d,
2H, J = 8.67 Hz), Ei.78 (s, 1H), 3.77 (s, 3H), 2.91 (2,
3H), 2.85 (s, 3H). ESMS: (M+H)' 305.2.
The compound of Preparation 17 (1.26 g, 4.14 mmol) was
combined with 10~ palladium/carbon (0.70 g) and
SUBSTITUTE SHEET (RULE 26)
Preparation 18


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palladium/black (0.15 g) in tetrahydrofuran(40 mL) and
the mixture shaken under a hydrogen atmosphere (38 psi)
in a Parr apparatus. After reduction was complete, the
catalyst was removed by filtration through celite and the
solution was immediately combined with 1,3-
dicyclohexylcarbod:iimide (0.82 g, mmol), 1-
hydroxybenzotriazole mono-hydrate (0.54 g, 4.0 mmol), the
product of Preparation lj, (1.50 g, 3.97 mmol), and
additional tetrahydrofuran (60. mI,). After stirring
overnight at ambient temperature, the mixture was
concentrated and the resulting residue slurried in ethyl
acetate and filters~d. The filtrate was concentrated and
the residue purified by silica gel chromatography
(chloroform/methanol) which provided 1.508 (57~) of the
desired product. MS: (M+H)+ 635.6. 1H NMR was
consistent with product. Anal. Calcd. for C3qHq6N6O6: C,
64.33; H, 7.30; N, 13.24. Found: C, 64.09; H, 7.09; N,
13.01.
Example 10
To a solution of the compound of preparation 18 (1.45 g,
2.29 mmol) stirring in dichloromethane (50 mL) at room
temperature was added triflouroacetic acid (15 mL). After
3 h, the reaction mixture was concentrated and the
residue treated with excess aqueous sodium bicarbonate.
SUBSTITUTE SHEET (RULE 26)


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The mixture was extracted with ethyl acetate and the
combined organic Extracts were dried over sodium sulfate
and concentrated. The residue was purified by flash
chromatography (sil.ica gel, chloroform/methanol) to give
0.73 g (60 ~) if the desired product as a yellow solid:
(60$. ESMS: (M+H)' 535.4. 1H NMR was consistent with
product . Anal . Ca:lcd. for CZ9H38N5O4 ~ 0 . 05 CHC13 : C, 64 . 54;
H, 7.09; N, 15.54. Found: C, 64.28; H, 6.70; N, 15.35.
The diastereomeric mixture (2.35 g) was resolved by HPLC
(8 x 15 cm Prochrom column packed with Kromasil CHI-DMP
chiral phase using an eluent mixture of 3A alcohol and
dimethylethylamine .in heptane to provide the individual
diastereomers in pure form (isomer 1, tR - 7.84 min),
isomer 2 (1.03 g, tR = 10.27 min). To a solution of isomer
2 (1.03 g, 1.93 mmol) in ethyl acetate was added a
saturated solution of hydrochloric acid in ethyl acetate.
The resulting solution was concentrated, treated with
diethyl ether and concentrated to provide 1.23 g of the
desired product as an off white solid: ESMS: (M+H)+ 535.3,
536.4. 'H NMR was consistent with product. Anal. Calcd.
for C29H38N5Oq~3.0 HC1: C, 59.08; H, 6.42; N, 13.05. Found:
C, 54.12; H, 6.38; PJ, 12.86.
The compound of preparation 17 (0.73 g, 2.38 mmol) was
combined with 10$ pa.lladium/carbon (0.50 g) and
SUBSTITUTE SHEET (RULE 26)
Preparation 19


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palladium/black (0.10 g) in tetradyrofuran(40 mL)and the
mixture shaken undo r hydrogen (38 psi) in a Parr
apparatus. After reduction was complete, the catalyst
was removed by fill=ration through celite and the
resulting solution was immediately combined with
dicyclohexylcarbodiimide (0.49 g, 2.38 mmol), 1-
hydroxybenzotriazol.e mono-hydrate (0.32 g, 2.37 mmol),
the product of Pre~~aration 1L (0.93 g, 2.39 mmol) and
additional tetrahyd~rofuran (60 mL). After stirring
overnight at ambient temperature, the mixture was
concentrated and the residue slurried in ethyl acetate
and filtered. The filtrate was concentrated and the
residue purified lay silica gel chromatography
(chloroform/methanol) to provide 0.76 g (50$) of the
desired product as an off white solid which was used
without further purification.
Example 11
To a solution of the compound of preparation 19 (0.74 g,
1.15 mmol) stirring .at room temperature in
dichloromethane (30 mL) was added triflouroacetic acid
(10 mL). After 2 h, the mixture was concentrated and the
residue treated with excess aqueous sodium bicarbonate.
The resulting mixturE~ was extracted with ethyl acetate
and the combined organic extracts were concentrated. The
SUBSTITUTE SHEET (RULE 26)


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residue was purified by flash chromatography (silica gel,
chloroform/methanol.) to provide 0.23 g (37$) of the
desired product: E;SMS: (M+H)+ 546.6. 1H NMR was
consistent with product. Anal. Calcd. for CZgH35N,Oq~0.25
CHC13: C, 61.05; H, 6.17; N, 17.04. Found: C, 61.41; H,
6.32; N, 16.52. The isomeric mixture (2.00 g) was
separated as described in Example 10 to provide 0.73 g of
isomer 1 (ta = 9.85 min) and 0.82 g of isomer 2 (tR =
12.87 min). To a solution of isomer 2 (0.82 g, 1.50
mmol) stirring in ei~hyl acetate and methanol was added a
saturated solution of hydrochloric acid in ethyl acetate.
The resulting mixture was concentrated to provide 0.84 g
of the desired product: ESMS: (M+H)+ 546.2, 547.3. 'H NMR
was consistent with product. Anal. Calcd. for CZgH35N,Oq~3.0
HC1: C, 53.18; H, 5.85; N, 14.97. Found: C, 53.73; H,
6.03; N, 14.04.
Preparation 20
O-
O
-"o
~/1 O
Reaction of (3,4-dimethoxyphenyl)acetic acid (30.0 g, 153
mmol) and p-toluenesulfonic acid (6.5 g, 33.8 mmol) in
absolute ethanol (201) mL) according to Preparation 1 gave
31.6 g (92~) of the desired product as a yellow oil: 1H-
NMR is consistent with structure; MS (ion spray) 225
(M+1 ) ; Anal . Calc' d for C12H1s04 : C, 64 . 27; H, 7 .19.
Found: C, 64.08; H, 7.07.
SUBSTITUTE SHEET (RULE 26j


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Preparation 21
O-
O
~~''0
Reaction of the compound of Preparation 20 (1.5 g, 6.7
mmol ) , N-bromosuccinimide ( 1 . 3 g, 7 . 4 mmol ) , 2, 2' -
azobis(2-methylpropionitrile) (0.2 g) in carbon
tetrachloride (30 m:L)as decribed in Preparation 2
provided 2.03 g (1000 of the desired product as a clear
oil: 1H-NMR is consistent with structure; Anal. Calc'd for
C~ZHISBr04: C, 47.54; H, 4.99. Found: C, 47.64; H, 5.17.
Preparation 22
0
i
o= N
N
~.,V.~_O
O\
0 l
Reaction of the product of Preparation 21 (13.3 g, 44
mmol), 4-nitroimidazole (5.0 g, 44 mmol) and sodium
hydride (2.1 g, 53 mmol) in tetrahydrofuran (400 mL) as
desribed in Preparation 3 provided 22.6 g (85$) of the
desired product as a tan oil. 'H-NMR is consistent with
structure; MS (ion spray) 334.1 (M-1); Anal. Calc'd for
C,sH1~N306'O.1CHC13: C, 52.23; H, 4.96; N, 12.10. Found: C,
52.55; H, 9.81; N, 11.85.
SUBSTITUTE SHEET (RULE 26)


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Preparation 23
N"J
~O
O ~O\
O-
O N N O
0
0 0
~~N
Hydrogenation of the compound of Preparation 22 (2.1 g,
6.3 mmol) with 10~ palladium on carbon (1.5 g) in
tetrahydrofuran (100 mL) followed by reaction with the
product of Preparation ld (2.4 g, 6.3 mmol), 1-
hydroxybenzotriazol~e ( 0 . 97 g, 6 . 9 mmol ) , 1- ( 3-
dimethylaminopropyl)-3-ethylcarbodiimide (1.43 g, 6.9
mmol) as described .in Preparation 4 gave 2.08 g (49~) of
the desired product as a red foam: 1H-NMR is consistent
with structure; MS (ion spray) 668.4 (M+1).
Preparation 24
"w0 - .O
I//
O
O N
O
O-
Reaction of the product of Preparation 23 (426814) (1.93
g, 2 . 9 mmol ) and lithium hydroxide ( 0 . 08 g, 3 . 5 mmol ) in
dioxane (50 mL)and water (25 mL) as described in
Preparation 5 provided 1.68 g (91$) of the desired
SUBSTITUTE SHEET (RULE 26)


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product as a tan foam: 'H-NMR is consistent with
structure; MS (ion spray) 640.3 (M+1); Anal. Calc'd for
C32H91N509: C, 60.08; H, 6.46; N, 10.95. Found: C, 60.31;
H, 6.75; N, 10.65.
Preparation 25
O N N O
,~ p
O N O
~N
~N ~NJ
~ \ o\
0
o-
Reaction of the product of Preparation 25 (426815) (0.8
g, 1.3 mmol), 4-methylpiperidine (0.16 mL, 1.3 mmol), 1-
hydroxybenzotriazole (0.2 g, 1.43 mmol) and 1-(3-
dimethylaminopropyl)-3-ethylcarbodiimide (0.3 g, 1.43
mmol) in dimethylformamide (20 mL) as desribed in
Preparation 6 provided 0.56 g (60$) of the desired
product as a tan foam: 1H-NMR is consistent with
structure; MS (ion spray) 721.5 (M+1); Anal. Calc'd for
CsaHSZNsOe : C, 63 . 31; H, 7 . 2 7 ; N, 11 . 6 6 . Found : C, 63 .18 ;
H, 7.30; N, 11.60.
Example 12
\O N N
/ O
O N
~N 2HC1
J
.~N N
O ~Ov
2 0 I O-
SUBSTITUTE SHEET (RULE 26)


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Reaction of the compound of Preparation 25 (0.5 g, 0.7
mmol and trifluoroacetic acid (2 mL) in dichloromethane
(6 mL) as described in Example 1 gave 0.4 g (83$) of the
desired mixture of isomers as a white solid: 'H-NMR is
consistent with structure; MS (ion spray) 621.6 (M+1);
Anal . Calc' d for Ca,3HqqN6O6'2 . 3hydrochloric acid: C, 56.25;
H, 6.62; N, 11.93. Found: C, 56.39; H, 6.33; N, 11.83.
Preparation 26
.O N N O
0
O N O
C %N
NJ
~ \ o\
0
o-
Reaction of the the product of Preparation 24 (0.8 g, I.3
mmol), dimethylamin.e hydrochloride (0.11 g, 1.3 mmol),
triethylamine (0.2 mL, 1.43 mmol), 1-hydroxybenzotriazole
( 0 . 2 g, 1 . 4 3 mmol ) and 1- ( 3-dimethylaminopropyl ) -3-
ethylcarbodiimide (0.3 g, 1.43 mmol) in dimethylformamide
(20 mL) as described in Preparation 6 gave 0.3 g (35$) of
the desired product as a tan foam: 'H-NMR is consistent
with structure: MS (ion spray) 667.4 (M+1); Anal. Calc'd
for C34HQ6N60e: C, 6I.25; H, 6.95; N, 12.60. Found: C,
60.83; H, 6.48; N, 12.45.
SUBSTITUTE SHEET (RULE 26)


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Example 13
r'~O N N
/ O
O N
N 2HC1
1
O
O-
Reaction of the product of Preparation 26 (0.28 g, 0.42
mmol) and trifluoroacetic acid (2 mL) in dichloromethane
(6 mL) as described in Example 1 gave 0.21 g (78$) of the
desired mixture of isomers as a white solid: 'H-NMR is
consistent with structure; MS (high res) calc'd for
CZ9H39N6O6: 567.2931. Found: 567.2938 . Anal . Calc' d for
CasHseNs~s'2hydrochloi-ic acid: C, 54.46; H, 6.30; N, 13.14.
Found: C, 54.67; H, 6.08; N, 13.00.
Preparation 27
C F3
Reaction of 4-trifluoromethylphenyl acetic acid (15.0 g,
73.4 mmol) and p-to.l.uenesulfonic acid (2.8 g, 14.7 mmol)
in absolute ethanol (100 mL) as described in Preparation
1 gave 16.3 g (95$) of the desired product as colorless
oil: 'H-NMR (d, DMSn) 1.18 (t, J = 7.0 Hz, 3H) , 3. 80 (s,
2H) , 4.10 (q, J = 7. 0 Hz, 2H) , 7 . 49 (d, J = 7 . 9 Hz, 2H) ,
7. 69 (d, J = 7. 9 Hz, 2H) ; MS (FD) 232 (M+) ; Anal. Calc' d
for CllHlF30z: C, 56.90; H, 4.77. Found: C, 56.81; H,
4.85.
SUBSTITUTE SHEET (RULE 26)


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CF3
Reaction of the product of Preparation of 27 (15.8 g,
68.0 mmol), N-bromosuccinimide (12.5 g, 70 mmol) and 48~
HBr (3 drops) in carbon tetrachloride (80 mL) as
described in Preparation 2 gave 19.8 g (94~) of the
desired product as a colorless oil: 1H-NMR (d, DMSO) 1.19
(t, J = 7.2 Hz, 3H), 4.15-4.25 (m, 2H), 6.07 (s, 1H),
7.78 (s, 4H) ; MS (FD) 309, 311 (M+) ; Anal. Calc' d for
C,lHIOBrF302: C, 42.47; H, 3.24. Found: C, 42.38; H, 3.13.
Preparation 29
a'
o= N:
N
O
C F~
Reaction of the prodvuct of Preparation 28 (51.8 g, 167
mmol), 4-nitroimidazole (18.8 g, 167 mmol), and potassium
carbonate (51 g, 368 mmol) in N,N-dimethylformamide (600
mL) as described Preparation 3 gave 21.7 g (38$) of the
desired product as a viscous orange oil: 'H-NMR (d, DMSO)
1.19 (t, J = 7.2 Hz, 3H), 4.26 (q, J = 7.2 Hz, 2H), 6.80
(s, 1H) , 7.76 (d, J =- 8.3 Hz, 2H) , 7. 83 (d, J = 8 .3 Hz,
2H) , 8. O1 (s, 1H) , 8.51 (s, 1H) ; MS (ion spray) 344
(M+1 ) ; Anal . Calc' d for C,9HIZFaN30q : C, 48 . 99; H, 3 . 52; N,
12.24. Found: C, 4_9.03; H, 3.74; N, 11.96.
SUBSTITUTE SHEET (RULE 26)
Preparation 28


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Preparation 30
O N N O
O
O N O
'~ N
O N
O
CF3
Hydrogenation of the product of Preparation 29 (8.5 g,
24.8 mmol) with 10~ palladium on carbon (6.0 g) in
tetrahydrofuran (70 mL) followed by coupling with the
product of Preparation ld (9.5 g, 24.8 mmol), 1-
hydroxybenzotriazole (3.7 g, 27.3 mmol) and 1-(3-
dimethylaminopropyl)-3-ethylcarbodiimide (5.6 g, 27.3
mmol)as described in Preparation 4 gave 12.8 g (77~) of
the desired product as a tan foam: 'H-NMR (d, DMSO) 1.17
(t, J = 7.2 Hz, 3H), 1.25-1.35 (m, 15H), 3.60 (m, 1H),
3.70 (m, 1H), 4.27 (q, J = 7.2 Hz, 2H), 4.44 (d, J = 2.6
Hz, 2H) , 4 . 60 (m, 1H) , 6. 63 (s, 1H) , 7 . 23-7. 30 (m, 7H) ,
7. 45 (m, 1H) , 7 . 58-7. 65 (m, 3H) , 7. 81 (d, J = 8 . 3 Hz,
2H), 10.25 (br s, 1H); MS (ion spray) 676.5 (M+1); Anal.
Calc'd for C33HaoFsNsO~,: C, 58.66; H, 5.97; N, 10.36.
Found: C, 58.58; H, 6.17; N, 10.27.
SUBSTITUTE SHEET (RULE 28)


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Preparation 31
y O N N
o ~ /\r
o~ o
/~N
J
O N
O
CFA
Reaction of the product of Preparation 30 (12.3 g, 18.2
mmol) and lithium hydroxide (0.52 g, 21.8 mmol) in
dioxane (100 mL) and water (75 mL) as described in
Preparation 5 gave 11.8 g (100$) of the desired product
as tan foam: 'H-NNtR (d, DMSO) 1. 20-1. 35 (m, 15 H) , 3 . 60
(m, 1H) , 3. 65 (m, 1H) , 4 . 45 (d, J = 2 . 6 Hz, 2H) , 4 . 60 (m,
1H), 6.46 (s, 1H), 7.15 (m, 1H), 7.20-7.35 (m, 6H), 7.42
(m, 1H), 7.57-7.65 (m, 3H), 7.79 (d, J - 8.3 Hz, 2H),
10.25 (br s, 1H); MS (ion spray) 648.9 (M+1); Anal.
Calc' d for C31H36F'3r~5O-7: C, 57. 41; H, 5. 60; N, 10. 81.
Found: C, 57.31; H, 5.59; N, 10.53.
Preparation 32
N N O
/ O
O N O
~N
~NJ
O
CFs
Reaction of the product of Preparation 31 (8.0 g, 12.3
mmol), 4-methylpiperi.dine (1.5 mL, 12.3 mmol), 1-
hydroxybenzotriazole (1.83 g, 13.5 mmol) and 1-(3-
dimethylaminopropyl)-3-ethylcarbodiimide (2.8 g, 13.5
SUBSTITUTE SHEET (RULE 26)


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mmol) in N,N-dimethylformamide (150 mL) as described in
Preparation 6 gave 7.33 g (81$) of the desired product as
a tan foam: 'H-NMR (d, DMSO) 0.78 (d, J = 6. 0 Hz, 1.5H) ,
0. 84 (d, J = 6. 0 Hz, 1. 5H) , 0. 95 (m, 1H) , 1. 25-1. 35 (m,
16H) , 1. 50-1 . 70 (m, 4H) , 2 . 65 (m, 1H) , 3 . 60 (m, 1H) , 3. 67
(m, 1H) , 3 . 80 (m, 1H) , 4 . 35-4 . 50 (m, 3H) , 4 . 60 (m, 1H) ,
6.88 (d, J = 9.8 Hz, 1H), 7.20-7.30 (m, 7H), 7.45 (m,
1H), 7.48-7.55 (m, 2H), 7.60 (m, 1H), 7.75-7.85 (m, 2H),
10.25 (br s, 1H); MS (ion spray) 729 (M+1); Anal. Calc'd
for C3~Hq~F3N6O6: C, Ei0.98; H, 6.50; N, 11.53. Found: C,
61.24; H, 6.44; N, 11.77.
Examples 14 and 15
\ ~~~.O N N
/ '/~ O
O N
C %N 2HC1
~N N J
O
CF3
Reaction of the product of Preparation 32 (7.0 g, 10.0
mmol) and trifluoroacetic acid (10 mL) in dichloromethane
(25 mL) as described in Example 1 gave 5.62 g (93$) of
the desired product (3.0 g) as a tan foam which was
purified by HPLC (8 :x 15 cm Prochrom column packed with
Kromasil CHI-DMP chi:ral phase with an eluent mixture of
3A alcohol and dimetllylethylamine in heptane) to give 1.5
g (45 $) of isomer 1 and 1.1 g (30 $) of isomer 2.
Example 14 (isomer 1) : 'H-NMR (d, DMSO) 0.25 (m, 1H) , 0.76
(d, J = 6. 4 Hz, 1.5H) , 0. 86 (d, J = 6.4 Hz, 1 . 5H) , 1.00
(m, 1H) , 1. 45-1. 70 (m, 8H) , 2 . 65-2 . 75 (m, 2H) , 3 . 15 (m,
1H), 3.65-3.80 (m, 3H), 4.40 (m, 1H), 4.51 (s, 2H), 4,75
(m, 2H) , 7 .10 (d, J =- 12. 8 Hz, 1H) , 7.20-7. 40 (m, 6H) ,
SUBSTITUTE SHEET (RULE 26)


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7.58 (d, J = 8.0 Hz, 1H), 7.67 (d, J = 8.0 Hz, 1H), 7.80-
7 . 90 (m, 2H) , 8 .10 (br s, 1H) , B . 20-8 . 35 (m, 3H) , 8 . 55
(d, J = 7.5 Hz, 1H), 10.95 (br s, 1H); t~ = 8.23 min; MS
( ion spray) 629. 3 (M+1 ) ; Anal . Calc. d for C32H;9F3N6O4 ~2HC1:
C, 54.78; H, 5.89; N, 11.98. Found: C, 54.85; H, 5.71;
N, 11.70.
Example 15 (isomer 2) : 'H-NMR (d, DMSO) 0.25 (m, 1H) , 0. 76
(d, J = 6. 4 Hz, 1 .5H) , 0. 86 (d, J = 6. 4 Hz, 1 .5H) , 1. 00
(m, 1H), 1.45-1.70 (m, 8H), 2.65-2.75 (m, 2H), 3.15 (m,
IO 1H), 3.65-3.80 (m, 3H), 4.40 (m, 1H), 4.51 (s, 2H), 4.75
(m, 2H) , 7. 10 (d, J' = 12. 8 Hz, 1H) , 7.20-7. 40 (m, 6H) ,
7 . 58 (d, J = 8 . 0 Hz, 1H) , 7 . 67 (d, J = 8 . 0 Hz, 1H) , 7 , 80-
7.90 (m, 2H), 8.10 (br s, 1H), 8.20-8.35 (m, 3H), 8.55
(d, J = 7. 5 Hz, 1H) , 10. 95 (br s, 1H) ; tR = 10. 77 min; MS
(ion spray) 629.3 (1H+1); Anal. Calc.d for
C32H3gF3N6Oq'2 .2HC1: C, 54 .22; H, 5. 86; N, 11. 85. Found: C,
59.15; H, 5.84; N, :11.64.
Preparation 33
N N- -O
/ O
O N
F , ~ ~N
N NJ
0 0
C F3
Reaction of the product of Preparation 31 (0.6 g, 0.93
mmol), 4-(4-fluorobe~azoyl)piperidine hydrochloride (0.23
g, 0 . 93 mmol ) , trietlzylamine ( 0 .15 mL, 10 . 2 mmol ) , 1-
hydroxybenzotriazole (0.14 g, 1.02 mmol) and 1-(3-
dimethylaminopropyl)--3-ethylcarbodiimide (0.21 g, 1.02
mmol)in dimethylformamide (30 mL) as described in
Preparation 6 providE:d in 0.35 g (45$) of the desired
SUBSTITUTE SHEET (RULE 26)


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product as a yello~r foam: 1H-NMR (d, DMSO) 1.25-1.35 (m,
15H), 1.40-1.50 (m, 2H), 1.75 (m, 1H), 1.85 (m, 1H),
2.85-3.00 (m, 2H), 3.55-3.75 (m, 3H), 3.90 (m, 1H), 4.40-
4.50 (m, 3H), 4.60 (m, 1H), 6.90 (m, 1H), 7.25-7.40 (m,
12H) , 7. 50-7. 60 (m,, 3H) , 8 . 03-8. 10 (m, 2H) , 10.20 (br s,
1H); MS (ion spray) 837.4 (M+1); Anal. Calc'd for
C43H48FqN6O7: C, 61.71; H, 5.78; N, 10.04. Found: C, 61.53;
H, 5.98; N, 9.95.
Example 16
\. ~ O N N
/ /~ O
O N
F / ~N 2HC1
N N...J
O O
CF3
Reaction of the product of Preparation 33 (0.34 g, 0.4
mmol) and trifluoroar_etic acid (2 mL) in dichloromethane
(6 mL) as described in Example 1 gave 0.2 g (63$) of the
desired product as a yellow solid: 'H-NMR (d, DMSO) 1.45-
1.65 (m, 6H), 1.75 (m, 1H), 1.85 (m, 1H), 2.85-3.05 (m,
2H), 3.25 (m, 1H), 3.60-4.00 (m, 7H), 4.40-4.55 (m, 3H),
4. 75 (m, 1H) , 7. 05 (d, J = 10. 6 Hz, 1H) , 7.25-7. 40 (m,
8H), 7.55-7.70 (m, 2H), 7.75-7.85 (m, 2H), 8.00-8.10 (m,
2H), 8.15-8.25 (m, 3H), 8.50 (d, J = 7.2 Hz, 1H), 10.75
(br s, 1H); MS (ion spray) 737.0 (M+1); Anal. Calc'd for
C3eHaoFaNsOs'2.4HC1: C, 55.37; H, 5.18; N, 10.20. Found: C,
55.39; H, 5.45; N, 10.07.
SUBSTITUTE SHEET (RULE 26)


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Preparation 34
N N O
O \
N O N I IO
~N
o ~NJ
' ~ \
C Fs
Hydrogenation of the product of Preparation 29 (1.75 g,
5.1 mmol) with 10'~ palladium on carbon (1.4 g) in
tetrahydrofuran (60 mL) followed by reaction with the
product of Preparation 1L (2.0 g, 5.1 mmol), 1-
hydroxybenzotriazole (0.76 g, 5.6 mmol) and 1-(3-
dimethylaminopropyl)-3-ethylcarbodiimide (1.16 g, 5.6
mmol) as described :in Preparation 4 gave 2.51 g (72~) of
the desired product as a tan foam: 'H-NMR (d, DMSO) 1.15-
1. 35 (m, 18H) , 3 . 05--3 .15 (m, 2H) , 4 . 25 (m, 2H) , 4 . 65 (br
s, 1H) , 6. 62 (s, 1H) , 6. 85 (m, 1H) , 6. 95-7. 08 (m, 2H) ,
7.20-7.30 (m, 2H), i.40-7.55 (m, 2H), 7.55-7.65 (m, 3H),
7.82 (d, J = 8.3 Hz, 2H), 10.20 (br s, 1H), 10.75 (br s,
1H) ; MS (ion spray) 685 (M+1) ; Anal. Calc'd for
C34H39F3N605-1H20: C, 58.11; H, 5.88; N, 11 .96. Found: C,
58.15; H, 5.59; N, 11.92.
Preparation 35
N N O
O ~\
O N O
C %N
O N
' ~ \
O
2 0 cF~
SUBSTITUTE SHEET (RULE 26)


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Reaction of the product of Preparation 34 (2.2 g, 3.2
mmol) and lithium hydroxide (0.1 g, 3.9 mmol) in dioxane
(50 mL) and water (25 mL) as described in Preparation 5
gave 2.1 g (1000 of the desired product as a tan foam:
'H-NMR (d, DMSO) , 1.. 15-1. 35 (m, 15H) , 3 . 05-3.15 (m, 2H) ,
4. 65 (br s, 1H) , 6. 97 (s, 1H) , 6. 90 (m, 1H) , 6. 98-7.10
(m, 2H), 7.20-7.30 (m, 2H), 7.40-7.55 (m, 2H), 7.57-7.64
(m, 3H) , 7. 80 (d, ~T = 8. 3 Hz, 2H) , 10. 20 (br s, 1H) ,
10.75 (br s, 1H), 1.3.80 (br s, 1H); MS (ion spray) 657.4
(M+1 ) ; Anal . Calc' cl for C32H35F3N6O6: C, 58 . 53; H, 5. 37; N,
12.80. Found: C, ~~9.28; H, 5.17; N, 12.65.
Preparation 36
N N O
~ O
N O "N O
~N
-~ ~NJ
v
0
C F3
Reaction of the product of Preparation 35 (0.7 g, 1,1
mmol), 4-methylpiperidine (0.13 mL, 1.1 mmol), 1-
hydroxybenzotriazole (0.17 g, 1.2 mmol) and 1-(3-
dimethylaminopropyl)-3-ethylcarbodiimide (0.26 g, 1.2
mmol) in N,N-dimethylformamide (30 mL) as described in
Preparation 6 provided 0.47 g (58$) of the desired
product as a tan foam: 1H-NMR (d, DMSO) 0.78 (d, J - 6.4
Hz, 1.5H), 0.86 (d, J = 6.3 Hz, 1.5H), 1.15-1.35 (m,
18H), 1.50-1.70 (m, 3H), 2.60-2.70 (m, 2H), 3.00-3.15 (m,
2H), 3.30 (m, 1H), 4.40 (m, 1H), 4.65 (m, 1H), 6.85-
6.95(m, 2H), 7.00-7.:10 (m, 2H), 7.17-7.30 (m, 2H), 7.40-
7.60 (m, 4H), 7.75-7.85 (m, 2H), 10.20 (br s, 1H), 10.75
(br s, 1H); MS (ion spray) 738.5 (M+1); Anal. Calc'd for
SUBSTITUTE SHEET (RULE 28)


CA 02340344 2001-02-12
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CseHasFsN,05'1H20: C, 60.39; H, 6.40; N, 12.97. Found: C,
60.18; H, 6.21; N, 12.99.
Examples 17 and 18
N
2HC1
N N,.
O
C F3
Reaction of the product of Preparation 36 (4.8 g, 6.5
mmol) and trifluoroacetic acid (16 mL) in dichloromethane
(40 mL) as described in Example 1 gave .2.0 g (44$) of the
desired mixture as a tan foam. Purification by HPLC (8 x
cm Prochrom colun~r~ packed with Kromasill CHI-DMP chiral
10 phase with an eluent. mixture of 3A alcohol (13$ by v),
dimethylethylamine (0.2$ by v) in heptane at a flow rate
of 250 mL/min) gave 0.5 g (12 $) of isomer 1 and 0.4 g (9
$) of isomer 2. Example 17. (isomer 1) 'H-NMR (d, DMSO)
0- 77 (d, J = 6. 5 Hz, 1. 5H) , 0. 87 (d, J = 6. 0 Hz, 1 . 5H) ,
15 1.00 (m, 1H), 1.32 (s, 3H), 1.50 (s, 3H), 1.50-1.70 (m,
2H) , 2. 72 (m, 1H) , 3. 00-3.30 (m, 4H) , 3 . 75 (m, 1H) , 4 . 05-
4 . 33 (m, 3H) , 4 . 20 (m, 1H) , 4 . 78 (m, 1H) , 6. 94 (m, 3H) ,
7.20 (s, 1H), 7.30-7.40 (m, 2H), 7.55-7.70 (m, 2H), 7.75-
8.00 (m, 4H), 8.05-8.15 (m, 2H), 8.50 (m, 1H), 10.86 (s,
1H) , 11. 05 (s, 1H) ; 1=k = 6. O1 min; MS (ion spray) 638.2
(M+1 ) . Example 18. (~~.somer 2) rH-NMR (d, DMSO) 0. 77 (d, J
- 6. 5 Hz, 1. 5H) , 0. 8',~ (d, J = 6. 0 Hz, 1 . 5H) , 1 . 00 (m,
1H), 1.32 (s, 3H), 1.50 (s, 3H), 1.50-1.70 (m, 2H), 2.72
(m, 1H), 3.00-3.30 (m, 4H), 3.75 (m, 1H), 4.05-4.33 (m,
3H), 4.20 (m, 1H}, 4.78 (m, 1H), 6.94 (m, 3H), 7.20 (s,
1H), 7.30-7.40 (m, 2H:), 7.55-7.70 (m, 2H), 7.75-8.00 (m,
;5U8STITUTE SHEET (RULE 28)


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4H), 8.05-8.15 (m, 2H), 8.50 (m, 1H), 10.86 (s, 1H),
11.05 (s, 1H); tR = 7.5 min; MS (ion spray) 638.2 (M+1).
Preparation 37
N O
~O
~N
r/
O N
.~ O N
O
C F~
A mixture of the product of Preparation 29 (11.1 8, 32.3
mmol) and 5$ palladium on carbon (1.7 g) in
tetrahydrofuran (100 mL) was hydrogenated at 60 psi at
room temperature using a Parr apparatus. After 1.5 h,
the resulting brown solution was filtered through celite
and concentrated to give 8.8 g (87~) crude oil which was
used without purification. To a mixture of the amine
stirring at O °C in tetrahydrofuran (20 mL) was added the
product of Preparation lj (10.6 g, 28.1 mmol) in
tetrahydrofuran (30 mL). To this mixture was added 1-
hydroxy-7-azobenzotriazole (4.0 g, 29.5 mmol) and 1,3-
dicyclohexylcarbodiimide (6.1 g, 29.5 mmol). The
solution was allowed to warm to room temperature and the
resulting mixture filtered after 3 days. The filtrate
was concentrated and subsequently purified by flash
chromatography (silica gel, 3.5~
methanol/dichlorometlzane) to provide 12.1 g (64$) of the
desired product as an orange solid: 1H-NMR (d, DMSO) 1.15
(t, J = 7 Hz, 3H), 1..18-1.32 (m, 15H), 1.35-1.70 (m, 4H),
3.23 (m, 2H) , 4 .19 (q, J = 7 Hz, 2H) , 4 . 31 (m, 1H) , 6. 58
(s, 1H) , 7. 00 (br s, 1H) , 7. 05-7.22 (m, 6H) , 7.41 (m,
1H), 7.52-7.58 (m, 3H), 7.75 (d, J = 8 Hz, 2H), 10.19 (br
SUBSTITUTE SHEET (RULE 26)


CA 02340344 2001-02-12
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s, 1H) ; MS ( ion spray) 674 . 7 (M+1 ) ; Anal . Calc' d for
C39H42F3N506: C, 60. E~1.; H, 6.28; N, 10. 39. Found: C, 60. 44;
H, 6.48; N, 10.36.
Preparation 3B
N~O
~O
'' ~~N
O N
~N
N -"J
O
r CFa
To a solution of tlhe product of Preparation 37 (12.0 g,
17. 8 mmol) stirring .in dioxane (20 ml) and water (20 ml) at
room temperature was added lithium hydroxide (0.84 g,
35.6 mmol). After 90 min with intermittent sonication,
the reaction was poured into a solution of sodium
bisulfate (12 g/50 mI~ Hz0) and brine (20 ml,) then extrar_ted
with ethyl acetate. The combined organic extracts were
dried over sodium sulfate, filtered, and concentrated to
provide 11.5 g (100$) of the desired product as a tan
solid: 'H-NMR (d, DMSO) 1.17-1.31 (m, 15 H), 1.40-1.70
(m, 4H), 2.45 (m, 2:H), 4.33 (m, 1H), 6.40 (s, 1H), 7.00
(m, 1H), 7.05-7.23 (m, 6H), ?.40 (m, 1H), 7.55-7.71 (m,
3H), 7.76 (d, J = 8 Hz, 2H), 10.25 (br s, 1H); MS (ion
spray) 646. 6 (M+1 ) ; Anal . Calc' d for C32H38F3N5O6~O . 7 H20:
C, 58.39; H, 6.03; N, 10.64. Found: C, 58.52; H, 6.01;
N, 9.87*.
SUBSTITUTE SHEET (RULE 26)


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Preparation 39
0
N~O
~O
N
O N
~N
O
C Fs
To a solution of the product of Preparation 38 (6.0 g,
9.3 mmol)stirring at 0°C in dimethylformamide was added
dimethylamine hydrochloride (0.76 g, 9.3 mmol),
diethylcyanophosphonate (1.41 mL, 9.3 mmol), and
triethylamine (1.29 mL, 9.3 mmol). After 30 min, a
second equivalent of dimethylamine hydrochloride, DECD
and triethylamine were added. After 30 min, the reaction
mixture was diluted with ethyl acetate (300 mL) and
washed with aqueous sodium bisulfate and brine. The
organic extract was dried over sodium sulfate, filtered,
and concentrated. The resulting crude material was
purified by radial chromatography (silica gel, 4$
methanol in dichloromethane) to give 4.7 g (75$) of the
desired product as a tan foam: 1H-NMR (d, CDC13) 1.25(s,
9H), 1.42(s, 6H), 1.60-1.80 (m, 4H), 1.90 (br s, 1H),
2.57 (m, 2H), 2.98 (s, 6H), 4.48 (m, 1H), 7.05-7.21 (m,
6H) , 7. 50 (m, 1H) , 7 . 62-7. 76 (m, 5H) , 8. 93 (br s, 1H) ,
10.93 (br s, 1H) ; MS (ion spray) 673.7 (M+1) .
SUBSTITUTE SHEET (RULE 26)


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Examples 19 and 20
N
~O
'' ~~N
O N
N
O
C F~
To the product of Preparation 39 (4.7 g, 7.0 mmol) was
stirred at room temperature in a saturated solution of
hydrochloric acid in glacial acetic acid (30 mL). After
90 min, the mixture was concentrated. The resulting
material diluted with ethyl acetate and extracted with
aqueous sodium bicarbonate. The organic extract was dried
over sodium sulfate, filtered, and concentrated to give
3.7 g (93~) of an orange solid. MS (ion spray) 573.4
(M+1). The diastereomers (3.4 g) were separated by chiral
chromatography using a Kromasil-CHI normal phase column
to provide 1.40 g (S6I~) of isomer 1 and 1.26 g (37~) of
isomer 2. The individual isomers were dissolved in a
saturated solution of hydrochloric acid in glacial acetic
acid (4 mL) and subsequently concentrated to provide the
desired products as tan solids:
Example 19 (isomer 1) 1H-NMR (d, DMSO) 1.41 (s, 3H), 1.42
(s, 3H), 1.51-1.73 (m, 4H), 2.53 (m, 2H), 2.82 (s, 3H),
2.84 (s, 3H), 4.39 (m, 1H), 6.91 (s, 1H), 7.10 (m, 3H),
7.18-7.29 (m, 3H), 7.55 (d, J = 8 Hz, 2H), 7.78 (d, J = 8
Hz, 2H) , 7.91 (br s,, 1H) , 8.15 (br s, 3H) , 8.38 (d, J -
7.5 Hz, 1H), 10.78 (br s, 1H); MS (ion spray) 573.4
SUBSTITUTE SHEET (RULE 26)


CA 02340344 2001-02-12
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-162-
(M+1 ) ; Anal . Calc .. d for C29H35F3N6~3'2 . 3HC1: C, 53 . 06; H,
5.73; N, 12.80. Found: C, 52.90; H, 5.66; N, 12.70.
Example 20. (isome=~ 2) 1H-NMR (d, DMSO) 1.42 (s, 6H),
1.51-1.73 (m, 4H), 2.53 (m, 2H), 2.82 (s, 3H), 2.84 (s,
3H), 4.39 (m, 1H), 6.91 (s, 1H), 7.10 (m, 3H), 7.18-7.29
(m, 3H), 7.55 (d, J = 8 Hz, 2H), 7.78 (d, J = 8 Hz, 2H),
7.91 (br s, 1H), 8.15 (br s, 3H), 8.38 (d, J = 7.5
Hz,lH), 10.78 (br s, 1H); MS (ion spray) 573.4 (M+1);
Anal. Calc.d for CZ<~H3SF3N6O3~2HC1: C, 53.96;H, 5.78; N,
13.02. Found: C, 53.84; H, 5.71; N, 12.93.
Preparation 40
CFA
O
O F
Reaction of (2-fluora-4-trifluoromethyl)phenylacetic acid
(20.0 g, 90 mmol) ar.~d p-toluenesulfonic acid (5.0, 26
mmol) in absolute ethanol (200 mL) as described in
Preparation 1 provided 22.5 g (1000 of the desired
product as a colorless oil: 1H-NMR is consistent with
structure; MS (FD) 250 (M+); Anal. Calc'd for CllHioF402:
C, 52.81; H, 4.03. Found: C, 52.94; H, 3.94.
Preparation 41
C F3
O
F
Br
Reaction of the product of Preparation 40 (16.8 g, 67
mmol), N-bromosuccinimide (12.3 g, 69 mmol) and 48~ HBr
(3 drops) in carbon tetrachloride (170 mL) as described
in Preparation 2 gave 22.05 g (1000 of the desired
product as a colorless oil: 1H-NMR is consistent with
SUBSTITUTE SHEET (RULE 26)


CA 02340344 2001-02-12
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structure; MS (FD) 328, 330 (M+); Anal. Calc'd for
CiiHsBrF402: C, 40.15; H, 2.?6. Found: C, 40.00; H, 2.77.
Preparation 42
o-
O ' N:
\~~ O
F3
Reaction of the product of Preparation 41 (21.4 g, 65
mmol), 4-nitroimidazole (8.8 g, 78 mmol) and potassium
carbonate (26.8 g, 195 mmol) in dimethylformamide (300
mL) as described in Preparation 3 gave 3.75 g (16~) of
the desired product as a tan oil: 1H-NMR is consistent
with structure; MS (ion spray) 362.2 (M+I); Anal. Calc'd
for C19H11FQN309: C, 46.55; H, 3.07; N, 11.63. Found: C,
47.13; H, 3.49; N, :L1.37.
Preparation 43
.'~. O N N O
O O
O
~~N
~-- O N F
O
CF3
I5 Reduction of the product of Preparation 42 (2.88 g, 8.0
mmol) with 10~ palladium on carbon (1.45 g) in
tetrahydrofuran (60 mL) follwed by coupling with the
product of Preparation ld gave (3.0 g, 8.0 mmol), 1-
hydroxybenzotriazole (1.2 g, 8.8 mmol) and 1-(3-
.SUBSTITUTE SHEET (RULE 26)


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dimethylaminopropy:l)-3-ethylcarbodiimide (8.8 mmol) as
described in Preparation 4 gave 2.85 g (51~) of the
desired product as a tan foam: 1H-NMR is consistent with
structure; MS (ion. spray) 694.4 (M+1); Anal. Calc'd for
C33H3gFqN5O~: C, 57.14; H, 5.67; N, 10.10. Found: C, 57.28;
H, 5.59; N, 10.09.
Preparation 44
N N O
I~ o
O N O
'' N
O F
' ~ \
O
CF3
Reaction of the product of Preparation 43 (2.64 g, 3,g
mmol) and lithium hydroxide (0.11 g, 4.6 mmol) in dioxane
(50 mL) and water (25 mL) as described in Preparation 5
gave 2.53 g (1000 of the desired product as a tan foam.
1H-NMR is consistent with structure; MS (ion spray) 664.4
(M+1).
Preparation 45
--~N N F
O
C F3
N N O
0
0
~N
.SUBSTITUTE SHEET (RULE 26)


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Reaction of the product of Preparation 44 (0.8 g, 1.2
mmol), 4-methylpiperidine (0.14 mL, 1.2 mmol), 1-
hydroxybenzotriazole (0.18 g, 1.32 mmol) and 1-(3-
dimethylaminopropyl)-3-ethylcarbodiimide (0.27 g, 1.32
mmol) in dimethylformamide (40 mL) as described in
Preparation 6 gave 0.63 g (70~) of the desired product
as a yellow foam: 1H-NMR is consistent with structure; MS
(ion spray) 747.4 (M+1).
Example 21
\ ~~ O N N
/ O
O N
2HC1
___CN N F
O
C F3
Reaction of the product of Preparation 46 (0.54 g, 0.72
mmol) and trifluoroacetic acid (2 mL) in dichloromethane
( 6 mL) as described in Example 1 gave 0 . 4 g ( 77$ ) of the
desired mixture of isomers as a white solid. 1H-NMR is
consistent with structure; MS (ion spray) 647.6 (M+1);
Anal. Calc'd for C,,2H3gFqN6Oq~2HCl: C, 53.41; H, 5.60; N,
11.68. Found: C, 53..34; H, 5.84; N, 11.65.
SUBSTITUTE SHEET (RULE 26)


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N N O-y(
o N° o
~'N
NJ F
1
r
CF3
Reaction of the product of Preparation 44 (0.8 g, 1.2
mmol), dimethylamine hydrochloride (0.1 g, 1.2 mmol),
triethylamine (0.19 mL, 1.3 mmol), 1-hydroxybenzotriazole
(0.18 g, 1.3 mmol) and 1-(3-dimethylaminopropyl)-3-
ethylcarbodiimide (0.27 g, 1.3 mmol) in dimethylformamide
(40 mL) as described in Preparation 6 gave 0.48 g (5g~)
of the desired product: 1H-NMR is consistent with
structure; MS (ion spray) 693.4 (M+1); Anal. Calc'd for
C33H40F4N6~6: C, 57.22; H, 5.82; N, 12.13. Found: C, 57,48;
H, 5.74; N, 12.02.
Example 22
N N
( / O
O'
2HC1
CF3
Reaction of the product of Preparation 46 (0.43 g, 0.62
mmol) and trifluoroacetic acid (2 mL) in dichloromethane
(6 mL) as described .in Example 1 gave 0.25 g (60~) of the
desired product as a mixture of diastereoisomers. 1H-NMR
SUBSTITUTE SHEET (RULE 26)
Preparation 46


CA 02340344 2001-02-12
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is consistent with structure; MS (ion spray) 593.9 (M+1);
Anal. Calc'd for C28H32F4N6Oq~2hydrochloric acid: C, 50.53;
H, 5.15; N, 12.62. Found: C, 50.25; H, 5.20; N, 12.35.
Preparation 47
F
O
S //~'' O
Reaction of 4-fluorophenylacetic acid (15.0 g, 97,0
mmol ) , p-toluenesu:Lfonic acid (2 . 0 g, 10 . 5 mmol ) and
absolute ethanol (100 mL) as described in Preparation 1
gave 15.4 g (87~) of the desired product as a colorless
oil: 1H-NMR (d, DMSO) 1.17 (t, J = 7.2 Hz, 3H) , 3.66 (s,
2H), 4.06 (q, J - '7.2 Hz, 2H), 7.10-7.20 (m, 2H), 7.25-
7 . 35 (m, 2H) ; MS (FD) 182 (M+) ; Anal . Calc' d for CloHmF02
C, 65.92; H, 6.09. Found: C, 65.67; H, 5.96.
Preparation 48
~~O
Reaction of the product of Preparation 47 (14.9 g, g2
mmol), N-bromosuccinimide (14.9 g, 84.5 mmol) and 48~ HBr
(4 drops) in carbon tetrachloride (80 mL) as described in
Preparation 2 gave 18.3 g (85~) of the desired product as
a colorless oil: 1H--NMR (d, DMSO) 1.19 (t, J - 7.2 Hz,
3H), 4.15-4.25 (m, 2H), 5.95 (s, 1H), 7.15-7.30 (m, 2H),
7.56-7.70 (m, 2H); 1KS (FD) 260, 262 (M+); Anal. Calc'd
for CloHioBrFOZ: C, 46.00; H, 3.96. Found: C, 46.10; H,
3.95.
SUBSTITUTE SHEET (RULE 26)


CA 02340344 2001-02-12
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Preparation 49
0
o.- N'
\~~ O
Reaction of the product of Preparation 48 (68 g, 260
mmol), 4-nitroimidazole (35.0 g, 312 mmol) and potassium
carbonate (108 g, 780 mmol) in dimethylformamide (300 mL)
as described in Preparation 3 gasve 39.8 g (52~) of the
desired product as an orange oil: 1H-NMR (d, DMSO) 1.83
(t, J = 7.2 Hz, 3H), 4.25 (q, J - 7.2 Hz, 2H), 6.66 (s,
1H), 7.25-7.35 (m, :?H), 7.55-7.65 (m, 2H), 7.95 (d, 1.13
Hz, 1H), 8.44 (d, J - 1.5 Hz, 1H); MS (ion spray) 294.2
(M+1) ; Anal. Calc'd for Cl3HizFNs04: C, 53.24; H, 4.12; N,
14.33. Found: C, 53.51; H, 4.07; N, 14.42.
Preparation 50
w0 N N O
( / O
O O
-,'' N
J
~O N
O
F
Reduction of the product of Preparation 49 (8.9 g, 30.3
mmol) with 10~ palladium on carbon (6.0 g) in
tetrahydrofuran (120 mL) follwed by coupling with the
product of Preparation 1d (11.4 g, 30 mmol), 1-
hydroxybenzotriazole (4.5 g, 33 mmol) and 1-(3-
SUBSTITUTE SHEET (RULE 26)


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dimethylaminopropyl)-3-ethylcarbodiimide (6.8 g, 33 mmol)
as described in Preparation 4 gave 10.8 g (58~) of the
desired product as a tan foam: 1H-NMR (d, DMSO) I.18 (t, J
= 7.2 Hz, 3H), 1.25-1.35 (m, 15H), 3.60 (m, 1H), 3.70 (m,
1H) , 4.25 {q, J ~ 7 .2 Hz, 2H) , 4 .44 (d, J = 2 . 6 Hz, 2H) ,
4.60 (m, 1H), 6.47 {s, 1H), 7.20-7.40 (m, 9H), 7.40-7.50
(m, 3H), 7.56 (s, 1H), 10.25 (br s, 1H); MS (ion spray)
626.1 (M+1); Anal, Calc'd for C32H4oFN50~: C, 61.43; H,
6.44; N, 11.19. Found: C, 61.63; H, 6.42; N, 11.26.
Preparation 51
N N O
0
O N
~'N
O N
O
F
Reaction of the product of Preparation 50 {10.5 g, 17.0
mmol) and lithium hydroxide (0.48 g, 20.4 mmol) in
dioxane (200 mL) and water (100 mL) as described in
Preparation 5 gave 10.1 g (1000 of the desired product
as a tan foam: 1H-NMR (d, DMSO) 1.25-1.40 (m, 15H), 3.35
(br s, 1H) , 3 . 60 (m, 1H) , 3 .70 (m, 1H) , 4.44 (d, J = 2. 6
Hz, 2H) , 4. 60 (m, 1:H) , 6.33 (s, 1H) , 7.20-7.35 (m, 9H) ,
7.40-7.50 (m, 3H), 7.56 (s, 1H), 10.20 (br s, 1H); MS
(ion spray) 598.5 (M+1) ; Anal. Calc'd for C3oH3sFNs0~: C,
60.29; H, 6.07; N, x:1.72. Found: C, 60.38; H, 6.29; N,
11.49.
SUBSTITUTE SHEET (RULE 26)


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Preparation 52
N N O
\ ~O
- O O
Reaction of product of Preparation 51 (9.2 g, 15.4 mmol),
4-methylpiperidine (1.83 mL, 15.4 mmol), 1-
hydroxybenzotriazole~ (2.3 g, 27 mmol) and 1-(3-
dimethylaminopropyl)-3-ethylcarbodiimide (3.5 g, 17 mmol)
in dimethylformamide~ (100 mL) as described in Preparation
6 gave 9.7 g (93~) of the desired product as a tan foam:
1H-NMR (d, DMSO) 0.76 (d, J = 6.1 Hz, 1.5H) , 0. 86 (d, J =
6.1 Hz, 1.5H), 1.00 (m, 1H), 1.20-1.40 (m, 15H), 1.45-
1.70 (m, 3H), 2.55-2.70 (m, 2H), 3.05 (m, 1H), 3.60 (m,
1H), 3.65-3.75 (m, 2H), 4.40 (m, 1H), 4.44 (d, J - 2.6
Hz, 2H), 4.60 (m, 1:H), 6.73 (d, J - 11.3 Hz, 1H), 7.15-
7.35 (m, 9H), 7.35-7.50 (m, 4H), 10.20 (br s, 1H). MS
( ion spray) 679 . 6 (:M+1 ) ; Anal . Calc' d for C36H4~FN6O6 : C,
63.70; H, 6.98; N, 12.38. Found: C, 63.44; H, 6.86; N,
12.22.
SUBSTITUTE SHEET (RULE 26)


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2HC1
Reaction of the product of Preparation 52 (9.7 g, 14.3
mmol) with trifluoroacetic acid (16 mL) in
dichloromethane (40 mL) as described in Example 1 gave
6.8 g (73~) of t:he desired product as a mixture of
diastereoisomers. The mixture (3.2 g) was purified by
HPLC (8 x 15 cm Prochrom column packed with Kromasil CHI-
DMP chiral phase with an eluent mixture of 3A alcohol and
IO dimethylethylamine in heptane) to give 0.8 g (24 ~) of
isomer 1 and 0.9 g (26 ~) of isomer 2 as white solids:
Example 23 (Isomer 1) . 1H-NMR (d, DMSO) 0.75 (d, J = 6.4
Hz, 1.5H) , 0. 88 (d, :J = 6.4 Hz, 1.5H) , 1. 20 (m, 1H) , 1.35
(m, 1H), 1.45-1.70 (m, 8H), 2.60-2.75 (m, 2H), 3.15 (m,
1H), 3.65-3.85 (m, :3H), 4.35 (m, 1H), 4.52 (s, 2H), 4.75
(m, 1H), 6.95 (d, ~T - 11.3 Hz, 1H), 7.20-7.49 (m, 9H),
7.45 (m, 1H), 7.52 (m, 1H), 8.05 (br s, 1H), 8.25 (m,
3H), 8.56 (m, 1H), 10.95 (br s, 1H); tR - 6.73 min; MS
(ion spray) 579,4 (M+1); Anal. Calc'd for
C31H3yFNs04'2HC1~0.2CHC.'13: C, 56.29; H, 6.24; N, 12.67.
Found: C, 56.47; H, 6.17; N, 12.24.
Example 24. (Isomer 2) 1H-NMR (d, DMSO) 0.75 (d, J = 6.4
Hz, 1.5H) , 0. 88 (d, J = 6.4 Hz, 1. 5H) , 1.10 (m, 1H) , 1.35
(m, 1H), 1.45-1.70 (m, 8H), 2.60-2.75 (m, 2H), 3.15 (m,
1H), 3.65-3.85 (m, 3H), 4.35 (m, 1H), 4.52 (s, 2H), 4.75
(m, 1H), 6.95 (d, J - 11.3 Hz, 1H), 7.20-7.49 (m, 9H),
SUBSTITUTE SHEET (RULE 26)
Example 23 and 24


CA 02340344 2001-02-12
WO 00/10565 PCT/US99/03525
-172-
7.45 (m, 1H), 7.52 (m, 1H), 8.05 (br s, 1H), 8.25 (m,
3H), 8.56 (m, 1H), 10.95 (br s, 1H); tR - 9.09 min; MS
(ion spray) 579.4 I;M+1) ; Anal. Calc'd for C31H3gFN6Oq~2HCl:
C, 57.14; H, 6.34; N, 12.90. Found: C, 57.17; H, 6.18;
N, 12.79.
Preparation 53
N"J
CN
/ ~ \
O
F
w0 N N O
O
O O
~~N
Reaction of the product of Preparation 51 (0.6 g, 1.0
mmol), pyrrolidine (0.08 mL, 1.0 mmol), 1-
IO hydroxybenzotriazole (0.15 g, 1.1 mmol) and 1-(3-
dimethylaminopropyl)-3-ethylcarbodiimide (0.23 g, 1.1
mmol) in dimethylformamide (20 mL)as described in
Preparation 6 gave 0.27 g (41~) of the desired product as
a white foam: 1H-NMR is consistent with structure; MS (FD)
650.5 (M+) ; Anal. Calc'd for C3qHq3FN6O6~0.6H20: C, 61.73;
H, 6.73; N, 12.70. Found: C, 61.98; H, 6.43; N, 12.66.
Example 25
N N
/ O
O
2HC1
~N N
O
F
SUBSTITUTE SHEET (RULE 26)


CA 02340344 2001-02-12
WO 00/10565 PCT/US99i03525
-173-
Reaction of the product of Preparation 53(0.2 g, 0.3
mmol) and trifluoroacetic acid (4 mL) in dichloromethane
(6 mL) as described in Example 1 gave 0.16 g (84~) of the
desired mixture of isomers as a yellow solid: 1H-NMR is
consistent with structure. MS (high res) calc'd for
C29H36F'N6~4: 551.2782. Found: 551.2790.
Preparation 54
\O N N O
/ O
O N O
~~N
\N N
' ~ \
O
F
Reaction of the product of Preparation 51 (1.0 g, 1,7
mmol), dimethylamine hydrochloride (0.14 g, 1.7 mmol),
triethylamine (0.26 mL, 1.9 mmol), 1-hydroxybenzotriazole
(0.26 g, 1.9 mmol) and 1-(3-dimethylaminopropyl)-3-
ethylcarbodiimide (0.4 g, 1.9 mmol) in dimethylformamide
(30 mL) as described in Preparation 6 gave 0.55 g (52~)
of the desired product as a tan foam: 1H-NMR is consistent
with structure; MS (ion spray) 625.4 (M+1); Anal. Calc'd
for C32H41FN6O6: C, 61.53; H, 6.61; N, 13.45. Found: C,
61.22; H, 6.33; N, 13.44.
SUBSTITUTE SHEET (RULE 26)


CA 02340344 2001-02-12
WO 00/10565 PGT/US99/03525
-174-
Example 26
\~ N N
O
i' '~ O
O N
~N 2HC1
~N
O
j
F
Reaction of the product of Preparation 54 (0.54 g, 0.86
mmol) and trifluoroacetic acid (2 mL) , dichloromethane (6
mL) as described i.n Example 1 gave 0.4 g (77~) of the
desired product a:~ a mixture of isomers: 1H-NMR is
consistent with structure. MS (ion spray) 525.4 (M+1);
Anal. Calc'd for C27H33FN6O6-2HC1: C, 54.27; H, 5.90; N,
14.06. Found: C, 5:3.11; H, 5.70; N, 13.58.
Preparation 55
N N' .O
O ~J.I(O
O N
F , ~ ~N
N NJ
O O
F
Reaction of the product of Preparation 51 (0.6 g, 1.0
mmol), 4-(4-fluorobe~nzoyl)piperidine hydrochloride (0.25
g, 1.0 mmol), triethylamine (0.16 mL, 1.1 mmol), 1-
hydroxybenzotriazole f0.15 g, 1.1 mmol) and 1-(3-
dimethylaminopropyl)--3-ethylcarbodiimide (0.23 g, 1_1
mmol) in dimethylformamide (40 mL) as described
Preparation 6 gave 0..42 g (53~) of the desired product as
a tan foam: 1H-NMR is consistent with structure; MS (ion
SUBSTITUTE SHEET (RULE 28)


CA 02340344 2001-02-12
WO 00/10565 PGT/US99/035Z5
-175-
spray) 787 . 4 (M+) ; Anal . Calc' d for C42H4aFzNEO-, : C, 63 . 83 ;
H, 6.17; N, 10.63. Found: C, 63.95; H, 5.90; N, 10.44.
Example 27
n 0 N N
i o
O N
~ 2HC1
F , ~N
\ (I N N.,
O O
F
Reaction of the product of Preparation 55(0.4 g, 0.5
mmol) with trifluoroacetic acid (2 mL) in dichloromethane
(6 mL) as described in Example 1 gave 0.32 g (82~) of the
desired product as a yellow foam: 1H-NMR is consistent
with structure. MS (high res) calc'd for C3~H42F2IV6ps:
687.3106. Found: 687.3103. Anal. Calc'd for
C3~H4oFzNs~s'2.4 HCl: C, 57.40; H, 5.52; N, 10.85. Found:
C, 57.56; H, 5.53; N,. 10.50.
Preparation 56
N N- .O
O I~IO
O N
I '~ N
~- O N
O
F
Reduction of the product of Preparation 4 (4.8 g, 16.0
mmol) with 10~ palladium on carbon (5.0 g) and
tetrahydrofuran (160 mL) followed by coupling with the
product of Preparation lj (6.0 g, 16.0 mmol), 1-
hydroxybenzotriazole (2.4 g, 17.6 mmol) and 1-(3-
SUBSTITUTE SHEET (RULE 26)


CA 02340344 2001-02-12
WO 00/10565 PCT/US99/03525
-176-
dimethylaminopropyl)-3-ethylcarbodiimide (3.6 g, 17.6
mmol ) as described in Preparation 4 gave 15 . 4 g ( 77$ ) of
the desired product as a tan foam: 1H-NMR (d, DMSO) 1.17
(t, J - 7.2 Hz, 3H), 1.23-1.45 (m, 15H), 1.45-1.57 (m,
6H), 7.16 (q, J - 6.8 Hz, 2H), 4.40 (m, 1H), 6.45 (s,
1H), 7.05 (m, 1H), 7.10-7.30 (m, 8H), 7.40-7.48 (m, 3H),
7.54 (s, 1H), 10..?0 (br s, 1H); MS (ion spray) 624.4
(M+1) ; Anal. Calc'c3 for C33H42FNsOs: C, 63.55; H, 6.79; N
11.23. Found: C, 63.83; H, 6.78; N, 11.38.
Preparation 57
Reaction of the product of Preparation 56 (14.8 g, 24.0
mmol) with lithium hydroxide (0.66 g, 29.0 mmol) in
dioxane (200 mL) and water (100 mL) as in described in
Preparation 5 gave 14.3 g (1000 of the desired product
as a tan foam: 1H-Nr~ (d, DMSO) 1.25-1.40 (m, I5H), 1.50-
1.75 (m, 6H), 4.40 (s, 1H), 6.60 (s, 1H), 7.05 (s, 1H),
7.10-7.30 (m, 8H), T.40-7.50 (m, 3H), 7.55 (s, 1H), 10.2
(br s, 1H), 13.63 (br s, 1H); MS (ion spray) 596.5 (M+1);
2 0 Anal . Calc' d for C31Ff3sFNsOs' 0 . ldioxane : C, 62 . 3 9 ; H, 6 . 47 ;
N, 11.59. Found: C, 62.16; H, 6.56; N, 11.28.
SUBSTITUTE SHEET (RULE 2B)


CA 02340344 2001-02-12
WO 00/10565 PC'T/US99/03525
-177-
Preparation 58
N N O
O
N I'O
C %N
N NJ
0
/ v
F
Reaction of the product of Preparation 57 (13.3 g, 23.1
mmol), 4-methylpiperidine (3 mL, 23.1 mmol), 1-
hydroxybenzotriazole (3.4 g, 25.4 mmol) and 1-(3-
dimethylaminopropyl)-3-ethylcarbodiimide (5.2 g, 25.4
mmol) in dimethyli=armamide (100 mL) as described in
Preparation 6 gave :14.4 g (93~) of the desired product as
a tan foam: 1H-NMR I;d, DMSO) 0.76 (d, J = 6.4 Hz, 1.5 H) ,
0.86 (d, J - 4.9 Hz, 1.5H), 1.00 (m, 1H), 1.25-1.45 (m,
17H), 1.45-1.75 (m, 8H), 2.60-2.80 (m, 2H), 3.75 (m, 1H),
4.30-4.45 (m, 2H), 6.71 (d J - 11.7 Hz, 1H), 7.05 (m,
1H), 7.10-7.30 (m, S~H), 7.30-7.45 (m, 3H), 10.15 (m, 1H);
MS (ion spray) 677.5 (M+1); Anal. Calc~d for C3~H.~9FN605:
C, 65.66; H, 7.30; N, 12.42. Found: C, 65.78; H, 7.19;
N, 12.44.
7Examples 28 and 29
N N
/ O
O N
~N
/~ \N-J 2hIC1
~N
O
F
Reaction of the product of Preparation 58(13.8 g, 20.4
mmol) with trif7_uoroacetic acid (16 mL) in
SUBSTITUTE SHEET (RULE 26)


CA 02340344 2001-02-12
wo oonos6s Pc~rmsmo3s2s
-178-
dichloromethane (40 mL) as described in Example 1 gave
10.5 g (89~) of the desired mixture as a tan foam. The
mixture ( 4 . 0 g ) was purified by HPLC ( 8 x 15 cm Prochrom
column packed with Kromasil CHI-DMP chiral phase with an
eluent mixture of 3A alcohol and dimethylethylamine in
heptane) to give 1.5 g (38 ~) of isomer 1 and 0.77 g
(20~) of isomer 2 as white solids: Example 28. (isomer 1)
~H-NMR (d, DMSO) 0.75 (t, J = 6.4 Hz, 1.5 H), 0.87 (t, J =
6 . 0 Hz, 1. 5 H) , 1.15 (m, 1H) , 1. 35 (m, 1H) , 1 . 45-1. 80 (m,
12H), 2.55-2.75 (m, 3H), 3.05 (m, 1H), 3.65-3.75 (m, 2H),
4.30-4.50 (m, 2H), 6.94 (d, J = 12 Hz, 1H), 7.10-7.20 (m,
2H), 7.20-7.40 (m, '7H), 7.45 (m, 1H), 7.55 (m, 1H), 8.08
(m, 1H), 8.15-8.30 (m, 3H), 8.44 (t, J - 7.2 Hz, 1H),
10.90 (br s, 1H); to - 6.62 min; MS (ion spray) 578.3
(M+1 ) ; Anal . Calc ~ d for C3zHaiFNsOs ~ 2 . 3HC1: C, 58 . 81; H,
6.61; N, 12.72. Found: C, 57.91; H, 6.55; N, 12.72.
Example 29. (isomer 2) 1H-NMR (d, DMSO) 0.75 (t, J = 6.4
Hz, 1.5 H), 0.87 (t, J - 6.0 Hz, 1.5 H), 1.15 (m, 1H),
1.35 (m, 1H), 1.45-1.80 (m, 12H), 2.55-2.75 (m, 3H), 3.05
(m, 1H), 3.65-3.75 (m, 2H), 4.30-4.50 (m, 2H), 6.94 (d, J
- 12 Hz, 1H), 7.10-7.20 (m, 2H), 7.20-7.40 (m, 7H), 7.45
(m, 1H), 7.55 (m, 1H), 8.08 (m, 1H), 8.15-8.30 (m, 3H),
8.44 (t, J = 7.2 Hz, 1H), 10.90 (br s, 1H); tR = 8.95 min;
MS (ion spray) 578.3 (M+1); Anal. Calc'd for
C32H41FN6~3'2.3HC1: C, 58.81; H, 6.61; N, 12.72. Found: C,
58.05; H, 6.64; N, 12.43.
Preparation 59
F
O ~ F
~~ O
Reaction of 3,4-diflworophenylacetic acid (25.0 g, 145
mmol) with p-toluenesulfonic acid (9.5 g, 49.5 mmol) in
SUBSTITUTE SHEET (RULE 2B)


CA 02340344 2001-02-12
WO 00/10565 PCT/US99/03525
-179-
absolute ethanol (:150 mL) as described in Preparation 1
gave 28.7 g (99~) of the desired product as a colorless
oil: 1H-NMR is consistent with structure; MS (?D) 201
(M+) .
Preparation 60
F
~a
Reaction of the product of Preparation 59 (10.0 g, 50.0
mmol, N-bromosuccin:imde (9.17 g, 51.5 mmol) and 48~ HBr
(4 drops) in carbon tetrachloride (40 mL) as in
Preparation 2 gave :12.0 g (86~) of the desired product as
a colorless oil which was used without further
purification: 1H-NMR is consistent with structure; MS
(ion spray) 278, 280 (M+1).
Preparation 61
0
o.-N'
~~~-O
F
Reaction of the product of Preparation 60 (10.5 g, 38
mmol), 4-nitroimidazole (5.2 g, 45.6 mmol) and potassium
carbonate (15.1 g, 114 mmol) in dimethylformamide (400
mL) as described in Preparation 3 gave 4.54 g (39~) of
the desired product as an orange oil: 1H-NMR is consistent
with structure; MS (i.on spray) 312.0 (M+1); Anal. Calc'd
for C13H11FzN3~a~0.2H20: C, 49.59; H, 3.65; N, 13.35.
Found: C, 49.58; H, 3.62; N, 13.09.
SUBSTITUTE SHEET (RULE 26)


CA 02340344 2001-02-12
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Preparation 62
O ~O
F
Reduction of the product of Preparation 61 (1.35 g, 4.3
mmol) with 10~ palladium on carbon (0.8 g) in
tetrahydrofuran (40 mL) followed by coupling with the
product of Preparation 1d (1.64 g, 4.3 mmol), 1-
hydroxybenzotriazole~ (0.7 g, 4.7 mmol) and 1- (3-
dimethylaminopropyl)--3-ethylcarbodiimide (1.04 g, 4.7
mmol) as described i.n Preparation 4 gave 1.9 g (69~) of
the desired product as a tan foam: 1H-NMR is consistent
with structure; MS (ion spray) 644 (M+1); Anal. Calc'd
for C3zH39FZN50~: C, 59.71; H, 6.11; N, 10.80. Found: C,
59.72; H, 6.04; N, 10.63.
Preparation 63
N N O
0
o r~
c
F
F
Reaction of the product of preparation 62 (1.9 g, 3.0
mmol) with lithium hydroxide (0.09 g, 3.6 mmol) in
dioxane (50 mL) and water (25 mL) as described in
SUBSTITUTE SHEET (RULE 26)


CA 02340344 2001-02-12
WO 00/10565 PCT/US99/035Z5
-181-
Preparation 5 gave 1.6 g (87~) of the desired product as
a tan foam: 1H-NMR is consistent with structure; MS (ion
spray) 616.4 (M+1).
Preparation 64
N N O
~O
0 N 0 O
-.'"N
N N
O
r
F
F
Reaction of the product of Preparation 63 (0.5 g, 0.8
mmol), 4-methylpiperidine (0.1 mL, 0.8 mmol), 1-
hydroxybenzotriazole (O.I2 g, 0.88 mmol) and 1-(3-
dimethylaminopropyl)-3-ethylcarbodiimide (0.18 g, 0.88
mmol) in dimethylformamide (40 mL) as described in
Preparation 6 gave 0.3 g (54~) of the desired product as
a white foam: 1H-NMR is consistent with structure; MS
( ion spray) 697 (M+1 ) ; Anal . Calc' d for C36H46FzN6p6; C,
62.06; H, 6.65; N, 12.06. Found: C, 61.82; H, 6.57; N,
11.96.
SUBSTITUTE SHEET (RULE 26)


CA 02340344 2001-02-12
WO 00/10565 PGT/US99/03525
-182-
Exempla 30
N N
_O
r o
O N
2HCI
N N
\ F
0
r
F
Reaction of the product of Preparation 64 (0.22 g, 0.3
mmol) and trifluoroacetic acid (2 mL) in dichloromethane
(6 mL) as described in Example 1 gave 0.2 g (1000 of the
desired mixture of isomers as a yellow foam: 1H-NMR is
consistent with structure; MS (ion spray) 597.5 (M+1);
Anal. Calc'd for C~lFi3aF2N6p4~2.2HC1; C, 55.01; H, 5.99; I~
12.42. Found: C, 55.16; H, 5.96; N, 12.20.
Preparation 65
N N, .O
.O ~ O
O
c~
F
Reaction of the product of Preparation 63 (0.5 g, O.g
mmol), pyrrolidine (0.07 mL, 0.8 mmol), 1-
hydroxybenzotriazole (0.12 mL, 0.88 mmol) and 1-(3-
dimethylaminopropyl)-3-ethylcarbodiimide (0.18 g, O,gg
mmol) in dimethylformamide (40 mL) as described in
Preparation 6 gave 0.25 g (42~) of the desired product as
SUBSTITUTE SHEET (RULE 26~


CA 02340344 2001-02-12
WO 00/10565 PCT/US99/035Z5
-183-
a tan foam: 1H-NMR is consistent with structure. MS (ion
spray) 669 . 4 (M+1 ) ; Anal . Calc' d for C3qHqzF2N6O~~ 0 . 7H20 : C,
59.94; H, 6.42; N, 12.33. Found: C, 59.96; H, 6.28; N
11.97.
E'Z 31
w
\ O N N
-i O
O
2HCi
CN N
O J \ F
F
Reaction of the product of Preparation 65 (0.2 g, 0.3
mmol) and trifluoroacetic acid (2 mL) in dichloromethane
(6 mL) as described in Example 1 gave 0.14 g (74~) of the
desired product as a yellow foam: 1H-NMR is consistent
with structure; MS (ion spray) 569.4 (M+1); Anal. Calc'd
for CZgH3qF2N6Oq~2.2HC1: C, 53.68; H, 5.62; N, 12.95.
Found: C, 53.83; H, 5.57; N, 12.37.
Preparation 66
O N N O
O
O N O
F ~ f /J
N
O p ~ \ F
F
Reaction of the product of preparation 63 (0.5 g, 0_g
mmol), 4-(4-fluorobenzoyl)piperidine hydrochloride (0.2
9~ 0.8 mmol), triethylamine (0.13 mL, 0.88 mmol), 1-
hydroxybenzotriazole (0.12 g, 0.88 mmol) and 1-(3-
dimethylaminopropyl)-3-ethylcarbodiimide (0.18 g, 0_g8
SUBSTITUTE SHEET (RULE 26)


CA 02340344 2001-02-12
WO 00/t0565 PCT/US99/03525
-184-
mmol) in dimethylformamide (40 mL) as described in
Preparation 5 gave 0.14 g (22~) of the desired product as
a white foam: 1H-NMR is consistent with structure; MS
( ion spray) 805 . 6 (M+1 ) ; Anal . Calc' d for Cq2H9~F3N6~~ : C,
62.68; H, 5.89; N, 10.44. Found: C, 62.45; H, 5.82; N,
10.40.
Example 32
O N N
-'' O
0 N
F ,. / ~N 2HC1
y ~ N
N
O p / \ F
F
Reaction of the product of Preparation 66 (0.14 g, 0.17
mmol) and trifluoroacetic acid (2 mL) in dichloromethane
(6 mL) as described in Example 1 gave 0.1. g (77~) of the
desired mixture of isomers as a yellow solid: 1H-NMR is
consistent with structure; MS (ion spray) 705.5 (M+1);
Anal. Calc'd for C3;H3gF3N6O5~2.1HC1: C, 56.88; H, 5.30; N,
10.76. Found: C, 56.64; H, 5.31; N, 10.30.
Preparation 67
Reaction of 2,4-difluorophenylacetic acid (20 g, 116
mmol) and p-toluenesulfonic acid (6.0 g, 31 mmol) in
absolute ethanol (150 mL) as described in Preparation 1
gave 22.1 g (95$) of the desired product as a colorless
SUBSTITUTE SHEET (RULE 26)


CA 02340344 2001-02-12
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-185-
oil which solidifies upon setting: -H-NMR is consistent
with structure; MS (FD) 200 (M+).
Preparation 68
Reaction of the product of Preparation 67 (21.4 g, 100
mmol), N-bromosuccinimide (19.6 g, 103 mmol) and 48~ HBr
(6 drops) in carbon tetrachloride (100 mL) as described
in Preparation 2 gave 27.9 g (1000 of the desired
product as a colorless oil: 1H-NMR is consistent with
structure; MS (FD) 278, 280 (M+); Anal. Calc'd for
CioH9BrF202: C, 43.04; H, 3.25. Found: C, 42.92; H, 3.15.
Preparation 69
o'
p _..~ N a
N F
\\V,.~ O
0
F
Reaction of the product of Preparation 68 (26.9 g, 96
mmol), 4-nitroimidazole (13.0 g, 115 mmol) and potassium
carbonate (40 g, 288 mmol) in dimethylformamide (150 mL)
as described in Preparation 3 gave 14.3 g (48~) of the
desired product as an arange oil: 1H-NMR is consistent
with structure; MS (ion spray) 312 (M+1); Anal. Calc'd
for C13H11F2N30q: C, 50.17; H, 3.56; N, 13.50. Found: C,
49.90; H, 3.56; N, 13.26.
SUBSTITUTE SHEET (RULE 26j


CA 02340344 2001-02-12
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Preparation 70
N, _O
r _o ~ ~~.~(o
0 N
~'. N
O N F
O
F
Reduction of the product of preparation 69(1.35 g, 4.3
mmol) with 10~ palladium on carbon (0.8 g) in
tetrahydrofuran (40 mL) followed by coupling with the
product of Preparation 1d (1.64 g, 4.3 mmol), 1-
hydroxybenzotriazole (0.7 g, 4.7 mmol) and 1-(3-
dimethylaminopropyl)-3-ethylcarbodiimide (1.04 g, 4.7
mmol) as desribed in Preparation 4 gave 1.52 g (55~) of
the desired product as a tan foam: 1H-NMR is consistent
with structure; MS (ion spray) 644.5 (M+1); Anal. Calc'd
for C32H3gF2N5O~: C, 59.71; H, 6.11; N, 10.88, rr~ound: C,
59.43; H, 5.97; N, 10.91.
Preparation 71
N N o
I~ o
O~N O
~N
J
O N F
O
F
Reaction of the product of preparation 70 (1.42 g, 2.2
mmol) with lithium hydroxide (0.07 g, 2.64 mmol) in
SUBSTITUTE SHEET (RULE 26)


CA 02340344 2001-02-12
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-187-
dioxane (50 mL) and water (25 mL) as described in
Preparation 5 gave 1.35 g (1000 of the desired product
as a tan foam: 1H-NMR is consistent with structure; MS
( ion spray) 616 , 3 (M+1 ) ; Anal . Calc' d for C3oH;;F2N50~ ; C,
58.33; H, 5.73; N, 11.38. Found: C, 57.71; H,r5.86; N,
10.80.
Preparation 72
\ O N N
f1 w~
'~N
F
Reaction of the product of Preparation 7I (0.6 g, 1.0
mmol), 4-methylpiperidine (0.12 mL, 1.0 mmol), 1-
hydroxybenzotriazole (0.15 g, 1.1 mmol) and 1-(3-
dimethylaminopropyl)-3-ethylcarbodiimide (0.23 g, 1.1
mmol) in dimethylformamide (30 mL) as described in
Preparation 6 gave 0.66 g (94~) of the desired product as
a tan foam: 1H-NMR is consistent with structure; MS (FD)
696 (M+) ; Anal . Calc' d for C36H96F2N6O6 : C, 62 . 05 ; H, 6 . 65;
N, 12.06. Found: C, 62.21; H, 6.48; N, 12.17.
Example 33
\ O N N
O
O
~~N
N~ 2HC1
__~~N F
O
F
SU8ST1TUTE SHEET (RULE 26)


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Reaction of the product of Preparation 72 (0.51 g, 0.73
mmol) and trifluoroacetic acid (2 mL) in dichloromethane
(6 mL) as in described in Example 1 gave 0.25 g (51~) of
the desired product as tan foam: 1H-NMR is consistent
with structure; MS (ion spray) 597.5 (M+1); Anal. Calc'd
for C31H3gF2N6Oq'2.2HCI: C, 55.01; H, 5.99; H, 12.42.
Found: C, 56.92; H, 5.98; N, 12.36.
Preparation 73
O N N O
O
O N O
~~N
~N N F
O
F
Reaction of the product of Preparation 71 (0.6 g, 1.0
mmol), pyrrolidine (0.8 mL, 2.0 mmol), 1-
hydroxybenzotriazole (0.15 g, 1.1 mmol) and 1-(3-
dimethylaminopropyl)-3-ethylcarbodiimide (0.23 g, 1.1
mmol); dimethylformamide (30 mL) as described in
Preparation 6 gave 0.4 g (58~) of the desired product as
a white foam: 1H-NMR is consistent with structure; MS
( ion spray) 669 . 5 (M+1 ) ; Anal . Calc' d for C32H42FZN6O6 : C,
61.07; H, 6.33; N, 12.57. Found: C, 60.84; H, 6.31; N,
12.32.
SUBSTITUTE SHEET (RULE 26~


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ale 34
O N N
_ O
2HC1
Reaction of the product of Preparation 73 (0.3 g, 0.45
mmol) and trifluoroacetic acid (2 mL) in dichloromethane
(6 mL) as described in Example 1 gave 0.21 g (70~) of the
desired product as a white foam: 1H-NMR is consistent with
structure; MS (ion spray) 569.4 (M+1); Anal. Calc'd for
CZgH3qF2N6Oq~2.3HC1: C, 53.38; H, 5.61; N, 12.88. Found: C,
53.59; H, 5.58; N, 12.42.
Preparation 74
O
N1
O=
N
N~
To a solution of the compound of Preparation 49 (17.0 g,
58.0 mmol) stirring at room temperature was added to
sodium hydroxide (125 mL of a 2N aqueous solution) along
with tetrahydrofuran (10 mL) and ethanol (10 mL). After
hydrolysis was complete, the mixture was cooled in an
bath and acidified to pH 2.75 with aqueous hydrochloric
SUBSTITUTE SHEET (RULE 26j


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acid and extracted with ethyl acetate. The combined
organic extracts were washed with water, dried over
sodium sulfate and concentrated to provide i5.0 g (gg~)
of the desired carboxylic acid. The crude material was
combined with aqueous N,N-dimethyl amine (40~, 9.0 mL,
71.8 mmol), 1-hydroxybenzotriazole hydrate (7.64 g, 56.6
mmol)and 1,3-dicyclohexylcarbodiimide (11.7 g, 56.6 mmol)
in tetrahydrofuran (150 mL). After 18 h, the mixture
was concentrated and the residue slurried in ethyl
acetate, filtered, and the filtrate concentrated.
Purification of the concentrate by flash crromatography
(silica gel, chloroform/methanol) provided 10.2 g (62~)
of the desired product: ESMS: (M+H)' 293.1. 1H NMR (300
MHz, DMSO-d6) 8 8.2.1 (d, 1H, J = 1. 51 Hz) 7, g0 (d~ lH~ J
1.13 Hz), 7.60-7.50 (m, 2H,), 7.38-7.25 (m, 2H), 6.88 (s,
1H), 2.92 (s, 3H), 2.86 (s, 3H). Anal. Calcd. for
C13H13N4~3: C, 53.43; H, 4.48; N, 19.17. Found: C, 53.43;
H, 4.71; N, 19.07.
N
The product of Preparation 74 (2.0 g (6.85 mmol) was
combined with 10~ palladium/carbon (1.80 g) and
palladium/black (0.20 g) in tetrahydrofuran(75 mL)and the
2 5 mixture shaken under a hydrogen atmosphere ( 3 8 ps i ) in a
Parr apparatus. After reduction was complete, the
SUBSTITUTE SHEET (RULE 26)
Preparation 75


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catalyst was removed by filtration through celite and the
resulting solution was immediately added to a solution of
1,3-dicyclohexylcarbodiimide (1.51 g, 7.3 mmol), 1-
hydroxybenzotriazole (1.0 g, 7.3 mmol), the product of
Preparation lj (2.77 g, 7.3 mmol) in tetrahydrofuran (50
mL) at room temperature. After 16 h, the mixture was
concentrated and the residmP ~lt'rrior~7 ,., ..~L._, _
then filtered. The filtrate was concentrated and
resulting crude product purified by flash chromatography
(silica gel, chloroform/methanol) which afforded 3.47 g
(81~) of the desired product: ESMS: (M+H)+ 623.5,
624.6. 1H NMR was consistent with product. Anal. Calcd.
for C33HqgN6OqFØ0'~? CHC13: C, 63.44; H, 6.94; N, 13.44.
Found: C, 63.04; H, 7.41; N, 11.93.
To a solution of the product of Preparation 75 (1.45 g,
2.29 mmol) stirring at room temperature in
dichloromethane (50 mL) was added triflouroacetic acid
(15 mL). After 3 hours, the mixture was concentrated
and the material treated with excess aqueous sodium
bicarbonate. The aqueous mixture was extracted with
ethyl acetate and the combined organic extracts
concentrate. The resulting residue was purified by flash
SUBSTITUTE SHEET (RULE 28)
Example 35


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chromatography (silica gel, chloroform/methanol) to
provide 1.55 g of the desired product: ESMS: (M+H)T
523.3. The isomeric mixture (3.44 g) was separated as
previously described in Example 7 to provide 0.98 g of
pure isomer 1 (tR = 7.94 min) and 0.81 g of isomer 2 (tR =
10.57 min). For isomer 2, 0.80 g (1.53 mmol) was
dissolved in ethyl acetate/methanol and treated with a
saturated solution of hydrochloric acid in diethyl ether.
The resulting mixture was concentrated to provide 0.90 g
(92~) of the desired product as a light tan solid: ESMS:
(M+H)+ 523.4, 524.5. 1H NMR was consistent with product.
.Anal. Calcd. for C~8H35NsQ3F~3.25 HC1: C, 52.46; H, 6.01;
N, 13.11. Found: C, 52.49; H, 6.23; N, 12.80.
Preparation 75
HBOC
O
-N
~I
N I
I ~ N~
F ~ O
The product of Preparation 74 (2.00 g, 6.85 mmol) was
combined with 10~ pa.lladium/carbon (1.80 g) and
palladium/black (0.20 g) in tetrahydrofuran 75 mL) and
the mixture shaken under hydrogen atmosphere (39 psi) in
a Parr apparatus. After reduction was complete, the
catalyst was removed by filtration through celite and the
amine/tetrahydrofuran solution was immediately combined
with 1,3-dicyclohexylcarbodiimide(1.41 g, 6.85 mmol), 1-
hydroxybenzotriazole mono-hydrate (0.93 g, 6.85 mmol),
the product of Preparation lj (2.60 g, 6.84 mmol) and
SUBSTITUTE SHEET (RULE 26)


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additional tetrahydrofuran (75 mL). After stirring
overnight at ambient temperature, the mixture was
concentrated and the residue slurried in ethyl acetate.
The filtrate was concentrated and the residue purified by
flash chromatography(silica gel, chloroform/methanol) to
provide 3.65 g (85~)of the desired product as a tan
solid: ESMS: (M+H)+ 625.4. 1H NMR was consistent with
product. Anal. Calcd. for C3iH41N606'0.03 chloroform: C,
61.17; H, 6.60; N, 13.34. Found: C, 61.25; H, 6.90; N,
12.69.
To a solution of the product of Preparation 75 (3.30 g,
5.3 mmol) stirring in dichloromethane (30 mL) at room
temperature was added triflouroacetic acid (10 m). After
3 h, the mixture was concentrated and the residue treated
with excess aqueous sodium bicarbonate. The resulting
mixture was extracted with ethyl acetate and the combined
organic extracts were washed with 1N aqueous sodium
hydroxide, dried over sodium sulfate, and concentrated.
The residue was purified by flash chromatography (silica
gel, chloroform/methanol) to provide 1.40 g (51~) of the
desired product as a light tan solid: ESMS: (M+H)+ 525.3.
1H NMR was consistent with product. Anal. Calcd. for
CZ~H33N6OqF'1.3 methanol: C, 60.03; H, 6.80; N, 14.84.
SUBSTITUTE SHEET (RULE 26)
Example 36


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Found: C, 60.19; H, 6.81; N, 14.56. The isomeric
mixture (3.20 g) was separated as previously described in
Example 7 to give 1.57 g of isomer 1 (tR = 7.57 min) and
0.88 g of isomer 2 (tR = 10.43 min). For isomer 2, O.gg g
(1.68 mmol) was dissolved in ethyl acetate and treated
with a saturated solution of hydrochloric acid in diethyl
ether. The resulting mixture was concentrated, washed
with diethyl ether to give 0.97 g of the desired product:
ESMS: (M+H)+ 525.4, 526.7. 1H NMR was consistent with
product. Anal. Calcd. for C25H33N609F~2.75 HC1: C, 51.73;
H, 6.07; N, 13.4:1. Found: C, 51.62; H, 5.74; N, 13.34.
Preparation 76
O
O
~O
Reaction of 4-ethoxyphenylacetic acid (23.5 g, 130 mmol)
and p-toluenesulfonic acid (4.0 g, 21 mmol) in absolute
ethanol (150 mL) as described in Preparation 1 gave 23.2
g (86~) of the desired product as a colorless oil: 1H-NMR
(d, DMSO) 1.17 (t, J = 7.2 Hz, 3H) , 1.31 (t, J = 7.2 Hz,
3H), 3.56 (s, 2H), 3.99 (q, J = 7.2 Hz, 2H), 4.05 (q, J =
7.2 Hz, 2H), 6.85 (d, J = 8.7 Hz, 2H), 7.14 (d, J = g.7
Hz, 2H); MS (ion spray) 209 (M+1); Anal. Calc'd for
C12H1603~ C, 69.21; H, 7.74. Found: C, 68.91; H, 7.55.
SUBSTITUTE SHEET (RULE 2B)


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To a solution of the product of Preparation 76 (53 g, 255
mmol) stirring in carbon tetrachloride (600 mL) at room
temperature was added 46.6 g (262 mmol) of N-
bromosuccinimide and 3.0 g (18.3 mmol) of 2,2'-azobis(2-
methylpropionitrile). The resulting reaction mixture was
heated to reflux. After 3.5 h, the solution was cooled
to room temperarure, filtered and concentrated. The
resulting oil was chromatographed on silica gel using
chloroform as eluant to afford 70.9 g (97~) of the
desired product as a colorless oil: 1H-NMR (d, DMSO) 1.17
(t, J = 7.2 Hz, 3H), 1.25-1.35 (m, 3H), 4.00-4.10 (m,
2H), 4.13-4.25 (m, 2H), 5.86 (s, 1H), 6.92 (d, J = 8.7
Hz, 2H), 7.47 (d, J = 9.0 Hz, 2H); MS (FD) 287, 289 (M+).
Preparation 78
0
o = N:
N
~~~ 0
p ~ w
0
Reaction of the product of Preparation 77 (11.4 g, 40
mmol), 4-nitroimidazole (4.5 g, 40 mmol) and potassium
carbonate (16.6 g, 120 mmol) in dimethylformamide (100
mL) as described in Preparation 3 gave 5.47 g (43~) of
the desired product as a yellow oil: 1H-NMR (d, DMSO) 1.18
SUBSTITUTE SHEET (RULE 26)
Preparation 77


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(t, J = 7.2 Hz, 3H), 1.29 (t, J = 7.2 Hz, 3H), 4.03 (q, J
- 7.2 Hz, 2H), 4.23 (q, J = 7.2 Hz, 2H), 6.54 (s, 1H),
6.70 (d, J = 8.7 Hz, 2H) , 7.42 (d, J = 8.7 Hz, 2H) , 7.90
(s, 1H), 8.34 (s, 1.H); MS (ion spray) 320.2 (M+1); Anal.
Calc'd for C15HI~N3O5: C, 56.42; H, 5.37; N, 13.16. Found:
C, 56.29; H, 5.17; N, 13.15.
Preparation 79
\ /~O V' .O
O
O
Reduction of the product of Preparation 78 (9.6 g, 30
mmol) with 10~ palladium on carbon (7.0 g) in
tetrahydrofuran (100 mL) followed by coupling with the
product of Preparation ld (11.5 g, 30 mmol), 1-
hydroxybenzotriazole (4.5 g, 33 mmol) and 1-(3-
dimethylaminopropyl)-3-ethylcarbodiimide (6.8 g, 33 mmol)
as described in Preparation 4 gave 9.9 g (50~) of the
desired product as a tan foam: 1H-NMR (d, DMSO) 1.17 (t, J
- 7.2 Hz, 3H), 1.25-1.40 (m, 18H), 3.58 (m, 1H), 3.70 (m,
1H), 4.02 (q, J = 7.2 Hz, 2H), 4.20 (q, J = 7.2 Hz, 2H),
4.44 (d, J = 3.4 Hz, 2H), 4.60 (m, 1H), 6.33 (s, 1H),
6.95 (d, J = 8.7 Hz, 2H), 7.15-7.35 (m, 9H), 7.43 (m,
1H), 7.51 (m, 1H), 10.2 (br s, 1H); MS (ion spray) 652.4
(M+1) ; Anal. Calc'd for C3qHq5N50g: C, 62.66; H, 6.96; N,
10.74. Found: C, 62.92; H, 7.00; N, 10.98.
SUBSTITUTE SHEET (RULE 26)


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Preparation 80
\ w0 O
Reaction of the product of Preparation 80 (9.7 g, 15.0
mmol) and lithium hydroxide (0.42 g, 18.0 mmol) in
dioxane (200 mL) and water (100 mL) as described in
Preparation 5 gave 9.4 g (1000 of the desired product as
a tan foam: 1H-NMR (d, DMSO) 1.25-1.40 (m, 18H), 3.60 (m,
1H), 3.68 (m, 1H), 4.02 (q, J - 7.2 Hz, 2H), 4.44 (d, J =
3.0 Hz, 2H), 4.60 (m, 1H), 6.19 (m, 1H), 6.95 (d, J = g.7
Hz, 2H), 7.28-7.35 (m, 9H), 7.40 (m, 1H), 7.51 (s, 1H),
10.2 (br s, 1H), 13.5 (br s, 1H); MS (ion spray) 624.5
(M+1 ) ; Anal . Calc' d for C43Hq1N50$ : C, 61 . 62 ; H, 6 . 63 ; N,
11.23. Found: C, 61.58; H, 6.92; N, 10.99.
Preparation 81
\ O N N O
0
O N O
~N
J
N
N
O
O
SUBSTITUTE SHEET (RULE 26)
O
--


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Reaction of the product of Preparation 80 (7.43 g, 12.0
mmol), 4-methylpiperidine (1.42 mL, 12.0 mmol), 1-
hydroxybenzotriazole (1.78 g, 13.2 mmol) and 1-(3-
dimethylaminopropyl)-3-ethylcarbodiimide (2.72 g, 13.2
mmol) in dimethylformamide (100 mL) as described in
Preparation 6 gave 6.4 g (76~) of the desired product as
a tan foam: 1H-NMR (d, DMSO) 0.74 (d, J = 6.4 Hz, 1.5 H),
0.87 (d, J = 6.0 Hz, 1.5H), 1.05 (m, 1H), 1.25-1.40 (m,
18H), 1.50-1.70 (m, 3H), 2.55-2.70 (m, 2H), 3.00 (m, 1H),
3.57 (m, 1H), 3.65-3.85 (m, 2H), 4.00-4.20 (m, 2H), 4.38
(m, 1H), 4.44 (d, .~ = 3.4 Hz, 2H), 4.60 (m, IH), 6.61 (d,
J = 12.0 Hz, 1H), 6.95-7.00 (m, 2H), 7.15-7.20 (m, 2H),
7.20-7.45 (m, 9H), 10.15 (br s, 1H); MS (ion spray) 705.5
(M+1 ) ; Anal . Calc' d for C3gH52N6O~ : C, 64 . 75 ; H, 7 . 44 ; N,
11.92. Found: C, 64.59; H, 7.21; N, 11.87.
Example 37 aad 38
N N
~O
r ~ o
O N
2HC1
~N N
O
J--
O
V
Reaction of the product of Preparation 81 (6.4, 9.1 mmol)
and trifluoroacetic acid (10 mL) in dichloromethane (25
mL) as described in Example 1 gave .4.71 g (77~) of the
desired mixture of diastereomers as a tan foam.
Resolution of the diastereomers (2.4 g) by HPLC (Kromsil
CHI-DMP chiral stationary phase, 3A
alcohol/dimethylethylamine/heptane) provided 200 mg (8
of isomer 1 and 0.8 g (31 ~) of isomer 2, both isolated
SUBSTITUTE SHEET (RULE 28)


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as white solids after acidification with hydrochloric
acid as described in Example 7:
Example 37. (Isomer 1) 1H-NMR (d, DMSO) 0.74 (d, J = 6.4
Hz, 1.5H), 0.88 (d, J = 6.0 Hz, 1.5H), 1.20 (m, 1H), 1.31
(t, J = 6.8 Hz, 3H), 1.45-1.70 (m, 8H), 2.60-2.70 (m,
2H), 3.05 (m, 1H), 3.65-3.80 (m, 3H), 4.00-4.20 (m, 3H),
4.37 (m, 1H), 4.52 (s, 2H), 4.75 (m, 1H), 6.80 (d, J -
13.2 Hz, 1H), 6.95-7.05 (m, 2H), 7.25-7.40 (m, 9H), 7.92
(br s, 1H), 8.20-830 (m, 3H), 8.53 (d, J = 7.2 Hz, 1H),
10.9 (br s, 1H) ; tR = 9.17 min; MS (ion spray) 605 (M+1) ;
Anal. Calc'd for C;3H4~N605~2HCI~0.1 CHC13: C, 58.45; H,
6.74; N, 12.74. Found: C, 58.64; H, 6.77; N, 12.36.
Example 38. (Isomer 2) 1H-NMR (d, DMSO) 0.74 (d, J = 6.4
Hz, 1.5H), 0.88 (d, J = 6.0 Hz, 1.5H), 1.20 (m, 1H), 1.31
(t, J = 6.8 Hz, 3H), 1.45-1.70 (m, SH), 2.60-2.70 (m,
2H), 3.05 (m, 1H), 3.65-3.80 (m, 3H), 4.00-4.20 (m, 3H),
4. 37 (m, 1H) , 4.52 (s, 2H) , 4 .75 (m, 1H) , 6. 80 (d, J =
13.2 Hz, 1H), 6.95-7.05 (m, 2H), 7.25-7.40 (m, 9H), 7.92
(br s, 1H), 8.20-8.30 (m, 3H), 8.53 (d, J = 7.2 Hz, 1H),
10.9 (br s, 1H); to = 12.68 min; MS (ion spray) 605 (M+1);
Anal. Calc'd for C;3H44N6OS~HC1: C, 59.35; H, 6.85; N,
12.98. Found: C, 59.62; H, 7.01; N, 12.71.
Preparation 82
N N' .O
~I'(O
/ ~ O O
O
~N
N'
O
O
Reaction of the product of Preparation 80 (0.9 g, 1.5
mmol), dimethylamine hydrochloride (0.13 g, 1.5 mmol),
SUBSTITUTE SHEET (RULE 26)


CA 02340344 2001-02-12
wo oonos6s Pcr~us99io3sZs
-200-
triethylamine (0.23 mL, 2.65 mmol), 1-
hydroxybenzotriazole (0.23 g, 1.65 mmol) and 1-(3-
dimethylaminopropyl)-3-ethylcarbodiimide (0.34 g, 1.65
mmol) in dimethylformamide (50 mL) as described in
Preparation 6 gave 0.46 g (47~) of the desired product as
a tan foam: 1H-NMR (d, DMSO) 1.25-1.35 (m, 18H), 2.90 (m,
6H), 3.57 (m, 1H), 3.67 (m, 1H), 4.03 (q, J = 7.2 Hz,
2H), 4.43-4.47 (m, 2H), 4.57 (m, 1H), 6.55 (m, 1H), 6.97
(d, J = 8.7 Hz, 2H), 7.15-7.45 (m, 11H), 10.16 (br s,
1H); MS (ion spray) 651.4 (M+1); Anal. Calc'd for
C39H96N607: C, 62. 7 5; H, 7.13; N, 12 . 91. Found: C, 62.55;
H, 6.84; N, 12.84"
Examples 39 aad 40
N N
~O
/~ O
O N
N 2HC1
N
O
O
Reaction of the product of Preparation 82 (0.44 g, 0.68
mmol)and trifluoroacetic acid (2 mL) in dichloromethane
(6 mL) as described in Example 1 gave 0.19 g (45~) of the
desired product as a tan foam. Resolution of the
diastereomers (90 mg, 0.14 mmol) by HPLC (Kromsil CHI-DMP
chiral stationary phase, 3A alcohol/dimethylethylamine
/heptane) provided 50 mg (50 ~) of isomer 1 and 27 mg (27
of isomer 2, both isolated as white solids after
acidification with hydrochloric acid as described in
Example 7:
Example 39. (isomer i): 1H-NMR (d, DMSO) 1.32 (t, J = 6.8
Hz, 3H) , 1. 50 (s, 6H) , 2. 86 (s, 3H) , 2. 90 (s, 3H) , 3 .70-
SUBSTITUTE SHEET (RULE 26)


CA 02340344 2001-02-12
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3.80 (m, 2H), 4.03 (q, J - 7.2 Hz, 2H), 4.52 (s, 2H),
4.75 (m, 1H), 6.76 (s, 1H), 7.00 (d, J - 8.7 Hz, 2H),
7.25-7.40 (m, 9H), 8.06 (m, 1H), 8.20-8.30 (m, 3H), 8.52-
8.60 (m, 1H), 11.00 (br s, 1H); tR - 7.70 min; MS (high
res ) calc' d for Cz9H3sNsOs : 551. 2982 . Founa: 551. 2987 .
Anal. Calc'd for C29H38N6~5~2.3 HC1~0.3ethyl acetate: C,
54.88; H, 6.51; N, 12.72. Found: C, 54.70; H, 6.49; N,
12.43.
Fxample 40. (isomer 2): 1H-NMR (d, DMSO) 1.32 (t, J = 6.8
Hz, 3H), 1.50 (s, 6H), 2.86 (s, 3H), 2.90 (s, 3H), 3.70-
3.80 (m, 2H), 4.03 (q, J = 7.2 Hz, 2H), 4.52 (s, 2H),
4.75 (m, 1H), 6.76 (s, 1H), 7.00 (d, J = 8.7 rz, 2H),
7.25-7.40 (m, 9H), 8.06 (m, 1H), 8.20-8.30 (m, 3H), 8.52-
8.60 (m, 1H), 11.00 (br s, 1H); tR = 9.09 min; MS (high
res) calc'd for Cz9H39N6Os: 551.2982. Found: 551.2976.
Anal. Calc'd for C2aH38N605~2.HC1~0.3 ethyl acetate: C,
55.18; H, 6.53; N, 12.79. Found: C, 55.01; H, 6.33; N,
12.54.
Preparation 83
N N O -7(
~~O
O
O
~'' N
CN
O
O
Reaction of the product of Preparation 80 (0.9 g, 1.5
mmol), pyrrolidine (0.13 mL, 1.5 mmol), 1-
hydroxybenzotriazole (0.23 g, 1.65 mmol) and 1-(3-
dimethylaminopropyl)-3-ethylcarbodiimide (0.34 g, 1.65
mmol) in dimethyiformamide (40 mL) as described in
Preparation 6 gave 0.7 g (74~) of the desired product as
SUBSTITUTE SHEET (RULE 26~


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a tan foam: 1H-NMR (d, DMSO) 1.25--1.40 (m, 18H), 1.70-1.90
(m, 4H), 2.95 (m, 1H), 3.30-3.40 (m, 2H), 3.55-3.70 (m,
3H), 4.03 (q, J = 7.2 Hz, 2H), 4.44 (d, J = 3.4 Hz, 2H),
4.57 (m, 1H), 6.34 (s, 1H), 6.97 (d, J = 8.7 Hz, 2H),
7.20-7.35 (m, 9H), 7.40-7.45 (m, 2H), 10.15 (br s, 1H);
MS (ion spray) 677.6 (M+1); Anal. Calc'd for
C3sHaaNs0~~0.2H20: C, 63.55; H, 7.17; N, 12.35. Found: C,
63.32; H, 6.96; N, 12.24.
Example 41
N N
/ _O ~ O
O
2HC1
CN
O
O
--
Reaction of the product of Preparation 83 (0.59 g, 0,9
mmol) and trifluoroacetic acid (2 mL) in dichloromethane
(6 mL) as described in Example 1 gave 0.36 g (64~) of
the desired product as a mixture of isomers: 1H-NMR (d,
DMSO) 1.32 (t, J = 6.8 Hz, 3H) , 1.45-1.60 (m, 6H) , 1. 65-
1.90 (m, 4H), 2.90 (:m, 1H), 3.25-3.45 (m, 2H), 3.65-3.75
(m, 3H), 4.02 (q, J = 6.8 Hz, 2H), 4.45-4.55 (m, 2H),
4.70-4.80 (m, 1H), 6.54 (s, 1H), 6.98 (d, J = 8.7 Hz,
2H), 7.20-7.40 (m, 9H), 8.05 (m, 1H), 8.20-8.30 (m, 3H),
8.54 (d, J = 7.2 Hz, 1H), 10.95 (br s, 1H); MS (high res)
calc'd for C31HQ1N6O5: 577.3138. Found: 577.3132. Anal.
Calc' d for C31H4oNsOs'2HC1: C, 57 . 32 ; H, 6 . 52 ; N, 12 . 94 .
Found: C, 57.46; H, 6.59; N, 12.91.
SUBSTITUTE SHEET (RULE 26)


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-203-
Preparation 84
O
O
~'"' O
Reaction of 4-butyloxyphenylacetic acid (10.0 g, 48 mmoI)
and p-toluenesulfonic acid (2.5 g, 13 mmol) in absolute
ethanol (100 mL) as described in Preparation 1 gave
11.04 g {98~) of the desired product as a colorless oil:
1H-NMR {d, DMSO) 0..94 (t, J = 7.4 Fiz, 3H), 1.18 (t, J =
7.0 Hz, 3H), 1.40--1.50 (m, 2H), 1.60-1.80 (m, 2H), 3.57
(s, 2H), 3.93 (q, J = 6.5 Hz, 2H), 4.08 (q, J = 7.3 Hz,
2H), 6.86 (d, J = 8.4 Hz, 2H), 7.15 (d, J = 8.4 Hz, 2H);
MS (ion spray) 237 (M+1); Anal. Calc'd for C19H2o03: C,
71.16; H, 8.53. Found: C, 71.33; H, 8.55.
Preparation 85
O
O
Br
To a solution of the product of Preparation 84, 6.0 g (25
mmol) in 100 mL of carbon tetrachloride was added 4.7 g
(25.8 mmol) of N-bromosuccinimide and 0.6 g of 2,2'-
azobis(2-methylpropionitrile). The reaction mixture was
heated to reflux. After 3.5 h, the mixture was cooled to
room temperature, filtered and concentrated. The
resulting oil was purified by flash chromatography
(silica gel, 3~ methanol/chloroform) to proved 6.9 g
SUBSTITUTE SHEET (RULE 26)


CA 02340344 2001-02-12
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(885) of the desired product as a colorless oil: 1H-NMR
(d, DMSO) 0.93 (t, J = 7.35 H, 3H) , 1.20 (t, J = 7.2 Hz,
3H), 1.40-1.50 (m, 2H), 1.60-1.80 (m, 2H), 3.95-4.05 (m,
2H), 4.10-4.15 (m, 2H), 5.87 (s, 1H), 6.93 (d, J = 8.7
Hz, 2H), 7.45 (d, J = 8.7 Hz, 2H); MS (FD) 314, 316 (M+);
Anal . Calc' d for C19H19Br03 ~ 0 . 5CHC13 : C, 52 . 54; H, 5 . 9g .
Found: C, 52.35; H, 5.84.
Preparation 86
o'
o._-_N~
\./ O
Reaction of the product of Preparation 85(5.82 g, 19.0
mmol), 4-nitroimidazole (2.1 g, 19.0 mmol) and potassium
carbonate (8.0 g, 57 mmol) in dimethylformamide (150 mL)
as described in Preparation 3 gave 3.5 g (53~) of the
desired product as a yellow oil: 1H-NMR (d, DMSO) 0.93
(t, J = 7.3 Hz, 3H), 1.19 (t, J = 7.0 Hz, 3H), 1.35-1.50
(m, 2H), 1.60-1.80 (m, 2H), 3.92-4.06 (m, 2H), 4.20-4.30
(m, 2H), 6.56 (s, 1H), 6.99 (d, J = 8.6 Hz, 2H), 7.44 (d,
J = 8.6 Hz, 2H) , 7.92 (s, 1H) , 8.37 (s, 1H) ; MS (ion
spray) 348 . 3 (M+1 ) ; Anal . Calc' d for C1~H21N305 : C, 58 . 78;
H, 6.09; N, 12.10. Found: C, 59.08; H, 6.21; N, 12.19.
SUBSTITUTE SHEET (RULE 26~


CA 02340344 2001-02-12
WO 00/10565 PCT/US99/03525
-205-
Preparation 87
N N
~O
.~ ~ O
O N
~'' N
.O N
O
O~
Reduction of the product of Preparation 86 (1.5 g, 4.3
mmol) with 10~ palladium on carbon (0.8g) in
tetrahydrofuran (40 mL) followed by coupling with the
product of Preparation ld (1.64 g, 4.3 mmol), 1-
hydroxybenzotriazole (0.7 g, 4.7 mmol) and 1-(3-
dimethylaminopropyl)-3-ethylcarbodiimide (1.04 g, 4.7
mmol) as described in Preparation 4 gave 1.1 g (38~) of
the desired product as a tan foam: 1H-NMR (d, DMSO) 0.92
(t, J = 7.5 Hz, 3H), 1.18 (t, J = 7.2 Hz, 3H), 1.25-1.40
(m, 15H), 1.40-1.50 (m, 2H), 1.60-1.75 (m, 2H), 3.60 (m,
1H), 3.70 (m, 1H), 3.95-4.00 (m, 2H), 4.20-4.25 (m, 2H),
4.45-4.48 (m, 2H), 4.57 (m, 1H), 6.35 (s, 1H), 6.97 (t, J
- 9.0 Hz, 2H), 7.15-7.35 (m, 9H), 7.40 (m, 1H), 7.50 (s,
1H), 10.20 (br s, 1H); MS (ion spray) 680.5 (M+2); Anal.
Calc'd for C36H49NSO8: C, 63.61; H, 7.27; N, 10.30. Found:
C, 63.53; H, 6.99; N, 10.54.
SUBSTETUTE SHEET (RULE 28)


CA 02340344 2001-02-12
WO 00/10565 PCT/US99/03525
-206-
J O
O
O~
Reaction of the praduct of Preparation 87 (1.1 g, 1.6
mmol) and lithium hydroxide (0.5 g, 1.92 mmol) in dioxane
(50 mL) and water (25 mL) as described in Preparation 5
gave 1.04 g (1000 of the desired product as a tan foam:
1H-NMR (d, DMSO) 0.95 (t, J = 7.5 Hz, 3H), 1.25-1.35 (m,
15H), 1.35-1.50 (m, 2H), 1.65-1.75 (m, 2H), 3.57 (m, 1H),
3.65 (m, 1H), 3.95 (t, J = 6.4 Hz, 2H), 4.57 (m, 1H),
6.19 (d, J = 1.5 Hz, 2H), 6.20 (s, 1H), 6.96 (d, J = 8.7
Hz, 2H), 7.10-7.35 (m, 9H), 7.40 (m, 1H), 7.50 (s, 1H),
10.20 (br s, 1H), 13.45 (br s, 1H); MS (ion spray) 652.5
(M+1 ) ; Anal . Calc' d for C32HasNsOa : C . 62 . 66 ; H, 6 . 96 ; N,
10.75. Found: C, 62.45; H, 7.07; N, 10.72.
J' O
0 00
o'~
SUBSTITUTE SHEET (RULE 26)
Preparation 88
Preparation 89


CA 02340344 2001-02-12
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-207-
Reaction of the product of Preparation 88 (1.0 g, 1.6
mmol), 4-methylpiperidine (0.19 mL, 1.6 mmol), 1-
hydroxybenzotriazole (0.24 g, 1.8 mmol) and 1-(3-
dimethylaminopropy:l)-3-ethylcarbodiimide (0.35 g, 1.8
mmol) in dimethylformamide (60 mL) as described in
Preparation 6 gave 0.57 g (48~) of the desired product
as a tan foam: 1H-NMR (d, DMSO) 0.75 (d, J = 6.0 Hz, 1H),
0.85-0.95 (m, 6H), 1.25-1.40 (m, 15H), 1.40-1.75 (m, 7H),
2.55-2.75 (m, 2H), 3.00 (m, 1H), 3.55 (m, 1H), 3.60-3.85
(m, 2H), 3.95-4.00 (m, 2H), 4.60 (m, 1H), 4.85-4.98 (m,
3H), 6.97 (d, J = 8.7 Hz, 1H), 6.90-7.00 (m, 2H), 7.15
(m, 1H), 7.20-7.45 (m, 10H), 10.15 (br s, 1H); MS (ion
spray) 733.5 (M+1) ; Anal. Calc'd for C4oH56N60~: C, 65.55;
H, 7.70; N, 11.47. Found: C, 65.44; H, 7.49; N, 11.59.
Examples 42 and 43
N N
l ,- _~ ~ o
O N
N 2HC1
--_~N. N
O
O~
Reaction of the product of Preparation 89 (0.55 g, 0.75
mmol} and trifluoroacetic acid (2 mL) in dichloromethane
(6 mL) as described in Example 1 gave 0.4 g (75~) of the
desired product as a mixture diastereomers. This material
was resolved by HPLC (Kromsil CHI-DMP chiral stationary
phase, 3A alcohol/dimethylethylamine/heptane) to provide
the desired diastereomers, both isolated as white solids
after acidification with hydrochloric acid as described
in Example 7:
SUBSTITUTE SHEET (RULE 26)


CA 02340344 2001-02-12
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-208-
Example 42. (isomer 1): 1H-NMR (d, DMSO) 0.75 (d, J = 6.4
Hz, 1H), 0.85-1.00 (m, 5H), 1.25-.1.40 (m, 2H), 1.40-1.50
(m, 2H), 1.50-1.60 (m, 6H), 1.60-1.75 (m, 4H), 2.60=2.70
(m, 2H), 3.00 (m, 1H), 3.60-3.75 (m, 3H}, 3.95-4.00 (m,
2H), 4.52 (s, 2H), 4.75 (m, 1H), 4.88 (m, 1H), 6.89 (d, J
- 14 Hz, 1H), 7.00-7.05 (m, 2H), 7.20-7.40 (m, 9H), g.10
(m, 1H), 8.20-8.30 (m, 3H), 8.60 (m, 1H), 11.02 (br s,
1H) ; tR = 5.90 min; MS (high res) calc'd for C35H99N6O5:
633.3764. Found: 633.3768. Anal. Calc'd for
C35HqgN6O5~2.3HC1: C, 58.66; H, 7.07; N, 11.73. Found: C,
58.59; H, 6.99; N, 11.46.
Example 43. (isomer 2): 1H-NMR (d, DMSO) 0.75 (d, J = 6.4
Hz, 1H), 0.85-1.00 (m, 5H), 1.25-1.40 (m, 2H), 1.40-1.50
(m, 2H), 1.50-1.60 (m, 6H), 1.60-1.75 (m, 4H}, 2.60=2.70
(m, 2H), 3.00 (m, 1H), 3.60-3.75 (m, 3H), 3.95-4.00 (m,
2H), 4.52 (s, 2H), 4.75 (m, 1H), 4.88 (m, 1H), 6.89 (d, J
- 14 Hz, 1H), 7.00-7.05 (m, 2H), 7.20-7.40 (m, 9H), 8.10
(m, 1H), 8.20-8.30 (m, 3H), 8.60 (m, 1H), 11.02 (br s,
1H) ; tR - 7 . 47 min; MS (high res ) calc ' d for C35H99N6O5
633.3764. Found: 633.3762. Anal. Calc'd for
C35H49N6~5'HC1: C, 59.57; H, 7.14; N, 11.91. Found: C,
59.74; H, 7.30; N, 11.72.
Reaction of 4-phenoxyphenylacetic acid (25.0 g, 110 mmol)
and p-toluenesulfonic acid (5.0 g, 26 mmol) in absalute
ethanol (150 mL) as in described in Preparation 1 gave
27.6 g (98~) of the desired product as a yellow oil: 1H-
SUBSTITUTE SHEET (RULE 2S)
Preparation 90


CA 02340344 2001-02-12
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NMR (d, DMSO) 1.18 (t, J - 7.2 Hz, 3H), 3.64 (s, 2H),
4.08 (q, J = 7.2 Hz, 2H), 6.90-7.00 (m, 4H), 7.13 (t, J =
7.5 Hz, 1H), 7.2.3 (d, J = 8.7 Hz, 2H), 7.40 (t, J - 5.7
Hz, 2H); MS (ion spray) 257.2 (M+1); Anal. Calc'd for
ClSHisOs- C, 74.98; H, 6.29. Found: C, 74.88; H, 6.31.
Reaction of the product of Preparation 90 (10.0 g, 39.0
mmol), N-bromosuccinimide (7.2 g, 40.2 mmol) and 48~ HBr
(4 drops) in carbon tetrachloride (40 mL) as described in
Preparation 2 gave 11.9 g (92~) of the desired product
as a colorless oil.: 1H-NMR (d, DMSO) 1.21 (t, J = 7..3 Hz,
3H), 4.15-4.30 (m, 2H), 5.94 (s, 1H), 6.95-7.15 (m, 4H),
7.20 (m, 1H), 7.40-7.50 (m, 2H), 7.52-7.70 (m, 2H); MS
(FD) 334, 336 (M+); Anal. Calc'd for C16H15BrOy 0.05CHC13:
C, 56.51; H, 4.45. Found: C, 56.85; H, 4.27.
preparation 92
0
p.-N
--.
0
Reaction of the product of Preparation 91 (10.9 g, 33.0
mmol), 4-nitroimidazole (4.5 g, 39.6 mmol) and potassium
carbonate (13.4 g, 99.0 mmol) in dimethylformamide (150
mL) as described in Preparation 3 gave 5.92 g (49~) of
the desired product as a yellow oil: 1H-NMR (d, DMSO)
SUBSTITUTE SHEET (RULE 26)
Preparation 91


CA 02340344 2001-02-12
wo oon os6s Pcrius99iossis
-210-
1.17 (t, J = 6.8 Hz, 3H) , 4.25 (q, J = 7.2 Hz, 2H) , 6.60
(s, 1H), 7.00-7.10 (m, 4H), 7.17 (t, J - 7.2 Hz, 1H),
7.43 (t, J = 6.0 Hz, 2H) , 7.53 (d, J = 6.8 Hz, 2H) , 7.94
(s, 1H), 8.41 (s, 1H); MS (ion spray) 368.2 (M+1); Anal.
Calc'd for ClyHl~N~05~0.15CHC13: C, 59.30; H, 4.49; N,
10.91. Found: C, 59.55; H, 4.73; N, 10.97.
Preparation 93
N N O
~O
I ~ o
O N
~'N
O N
O
O
Reaction of the product of Preparation 92 (1.58 (4.3
mmol) with 10~ palladium on carbon (0.8 g) in
tetrahydrofuran (70 mL) followed by coupling with the
product of Preparation 1d (1.64 g, 4.3 mmol), 1-
hydroxybenzotriazole (0.7 g, 4.7 mmol) arid 1-(3-
dimethylaminopropyl)-3-ethylcarbodiimide (1.04 g, 4.7
mmol) as described in Preparation 4 gave 1.92 g (62~) of
the desired product as a tan foam: 1H-NMR (d, DMSO) 1.20
(t, J = 7.2 Hz, 3H), 1.25-1.35 (m, 15H), 3.57 (m, 1H),
3.70 (m, 1H), 4.25 (q, J = 7.2 Hz, 2H), 4.45-4.47 (m,
2H), 4.60 (m, 1H), 6.43 (s, 1H), 7.00-7.10 (m, 4H), 7.20
(m, 1H), 7.25-7.35 (m, 6H), 7.35-7.45 (m, 6H), 7.55 (s,
1H), 10.20 (br s, 1H); MS (ion spray) 700.7 (M+1); Anal.
Calc' d for C38H45N5~H : C, 65 . 22 ; H, 6 . 48; N, 10 . 01. Found:
C, 65.12; H, 6.43; N, 9.87.
SUBSTITUTE SHEET (RULE 28)


CA 02340344 2001-02-12
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Preparation 94
N N O
-O
/ ~ O
O N O
/ --~" N
J
O N
O
O
Reaction of the product of Preparation 93 (1.72 g, 2.5
mmol) and lithium hydroxide (0.07 g, 3.0 mmol) in dioxane
(50 mL) and water (25 mL) as described in Preparation 5
gave 1.68 g (1000 of the desired product as a tan foam:
1H-NMR (d, DMSO) 1.25-1.40 (m, 15H), 3.60 (m, 1H), 3.70
(m, 1H), 4.45-4.50 (m, 2H), 4.57 (m, 1H), 6.25 (s, 1H),
7.00-7.07 (m, 4H), 7.15-7.35 (m, 8H), 7.35-7.45 (m, 5H),
7.55 (s, 1H), 10.20 (br s, 1H), 13.55 (br s, 1H); MS (ion
spray) 672 . 6 (M+1 ) ; Anal . Calc' d for C36H41N5O8 : C, 64 . 37 ;
H, 6.15; N, 10.43. Found: C, 64.56; H, 6.37; N, 10.23.
Preparation 95
N N O
_O
O N O O
'N
CN N
o / ~ i
O
Reaction of the product of Preparation 94 (0.45 g, 0.67
mmol), pyrrolidine (0.07 mL, 0.67 mmol), 1-
SUBSTITUTE SHEET (RULE 28)


CA 02340344 2001-02-12
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-212-
hydroxybenzotriazole (0.1 g, 0.74 mmol) and 1-(3-
dimethylaminopropyl)-3-ethylcarbodiimide (0.15 g, 0.74
mmol)in dimethylformamide (30 mL) as described in
Preparation 6 gave 0.22 g (45~) of the desired product
as a white foam: 7H-NMR (d, DMSO) 1.20-1.40 (m, 15H),
1.65-1.90 (m, 4H), 3.05 (m, 1H), 3.25-3.45 (m, 2H), 3.55-
3.75 (m, 3H), 4.45-4.50 (m, 2H), 4.60(m, 1H), 6.43 (s,
1H), 7.05 (t, J = 8.7 Hz, 3H), 7.20 (m, 1H), 7.25-7.30
(m, 7H), 7.35-7.50 (m, 7H), 10.20 (br s, 1H); MS (ion
spray) 725.7 (M+1); Anal. Calc'd for C4oH4aNs0~: C, 66.28;
H, 6.68; N, 11.59. Found: C, 66.42; H, 6.68; N, 11.59.
Example 44
2HC!
CN N
0
0
N N
~O
/ O
O
'' N
Reaction of the product of Preparation 95 (0.22 g, 0.3
mmol) and trifluoroacetic acid (2 mL) in dichloromethane
(6 mL) as described in Example 1 gave 0.2 g (1000 of the
desired mixture of isomers: 1H-NMR (d, DMSO) 1.45-1.55 (m,
6H), 1.70-1.90 (m, 4H), 2.95 (m, 1H), 3.25-3.45 (m, 2H),
3.50-3.90 (m, 3H), 4.45-4.55 (m, 2H), 4.75 (m, 1H), 6.60
(m, 1H), 7.00-(m, 3H), 7.20 (m, 1H), 7.25-7.50 (m, 12H),
7.98 (m, 1H), 8.15-8.30 (m, 3H), 8.52 (t, J = 7.6 Hz,
1H), 10.88 (br s, 1H); MS (ion spray) 625.4 (M+1); Anal.
Calc'd for C35H4oNsOs'2HC1; C, 60.26; H, 6.07; N, 12.05.
Found: C, 60.02; H, 6.01; N, 11.81.
SUBSTITUTE SHEET (RULE 26)


CA 02340344 2001-02-12
WO 00/10565 PCT/US99/03525
-213-
N \ N' .O
~I.I(O
O
O N O
/ ~'.N
J
~N N
o / ~ i
~I
0
Reaction of the product of Preparation 94 (0.6 g, 0.9
mmol), 4-methylpiperidine (0.1 mL, 0.9 mmol), 1-
hydroxybenzotriazole (0.14 g, 1.0 mmol) and 1-(3-
dimethylaminopropyl)-3-ethylcarbodiimide (0.2 g, 1.0
mmol)in dimethylformamide (30 mL) as described in
Preparation 6 gave 0.4 g (59~) of the desired product as
a white foam: 1H-NMR is consistent with structure; MS
( ion spray) 753 . 5 (M+1 ) ; Anal Calc' d for C92HSZN60~ : C,
67.00; H, 6.96; N, 11.16. Found: C, 66.73; H, 6.91; N,
11.04.
Example 45
..
2HC1
I
O
Reaction of the product of Preparation 96 (0.34 g, 0.45
mmol) and trifluoroacetic acid (2 mL) in dichloromethane
(6 mL) as described i.n Example 1 gave 0.27 g (83~) of
SUBSTITUTE SHEET (RULE 26)
Preparation 96


CA 02340344 2001-02-12
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-214-
the desired mixture of isomers as a white solid: 1H-NMR
is consistent with structure; MS (high res) calc'd for
C3~H4sNs~s : 653 . 3451. Found: 653 . 3446 .
Preparation 97
0
/ \
\ /
Reaction of biphenylacetic acid (25.2 g, 119 mmol) and p-
toluenesulfonic acid (3.3 g, 17 mmol) in absolute ethanol
(250 mL) as described in Preparation 1 gave 25.4 g (89~)
of the desired product as a yellow oil: 1H-NMR is
consistent with structure; MS (FD) 240.1 (M+); Anal.
Calc'd for C16H16~2: C, 79.97; H, 6.71. Found: C, 79.75;
H, 6.59.
Preparation 98
O
O
Br
Reaction of the product of Preparation 97 (18.0 g, 75.0
mmol), N-bromosuccinimide (13.7 g, 77.25 mL) and 48~ HBr
(4 drops) in carbon tetrachloride (80 mL) as described in
Preparation 2 gave 22.56 g (94~) of the desired product
as a yellow oil: iH-NMR is consistent with structure; MS
(FD) 318, 320 (M+}; Anal. Calc'd for.
ClsHlsBrOy 0.05Chydrochloric acid3: C, 60.21; H, 4.74.
Found: C, 59.50; H, 4.75.
SUBSTITUTE SHEET (RULE 28)


CA 02340344 2001-02-12
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Preparation 99
O
O=
,O
To a slurry of sodium hydride (2.42 g, 60.5 mmol)
stirring in dimethylformamide (200 mL) at room
temperature was added 4-nitroimidazole (6.9 g, 60.5
mmol). After 10 min, the product of Preparation 98
(17.62 g, 55.0 mmol) was added. After 16 h, the reaction
mixture was concentrated and the. residue was slurried in
ethyl acetate then filtered. The resulting oil was
partitioned between ethyl acetate and water then
extracted with ethyl acetate. The combined organic
extracts were washed with brine, dried over sodium
sulfate, filtered and concentrated. The resulting oil
was absorbed onto silica gel and purified by flash
chromatography (silica gel, 30-50~ ethyl acetate/hexanes)
to yield 12.0 g (62g) of the desired product as a yellow
viscous oil: 1H-NMR is consistent with structure; MS (FD)
351 (M+) .
Preparation 100
N N O
~~O
O
O
i
N
/ 1 /
SUBSTITUTE SHEET (RULE 26)


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-216-
Reduction of the product of Preparation 99 (2.0 g, 5.8
mmol) under a hydrogen atmosphere with 10~ palladium on
carbon (0.8 g) and tetrahydrofuran (70 mL) followed by
coupling with the product of Preparation ld (2.2 g, 5.8
mmol), 1-hydroxybenzotriazole (0.86 g, 6.4 mmol) and 1-
(3-dimethylaminopropyl)-3-ethylcarbodiimide (1.3 g, 6.4
mmol) as described in Preparation 4 gave 0.7 g (18~) of
the desired product as a tan foam: 1H-NMR is consistent
with structure; MS (FD) 683 (M+); Anal. Calc'd for
C38HQSN50~: C, 66.75; H, 6.63; N, 10.34. Found: C, 66.79;
H, 6.48; N, 10.32.
Preparation 101
N N O
O
/ O
O
O
Reaction of the product of Preparation 100 (0.7 g, 1.0
mmol ) and lithium hydroxide ( 0 . 03 g, 1. 2 mmol ) in dioxane
(20 mL) and water (10 mL) as described in Preparation 5
gave 0 . 66 g ( 1000 of the desired product as a tan foam:
1H-NMR is consistent with structure; MS (FD) 656 (M+);
Anal. Calc'd for C36HQ1N50~: C, 65.94; H, 6.30; N, 10.68.
Found: C, 65.90; H, 6.37; N, 10.42.
SUBSTITUTE SHEET (RULE 26)


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Preparation 102
N N' -O
/ _O ~ ~I.I(O
O N O
O
Reaction of the product of Preparation 101 (0.7 g, 1.1
mmol) withy 4-methylpiperidine (0.13 mL, 1.1 mmol), 1-
hydroxybenzotriazole (0.17 g, 1.2 mmol) and 1-(3-
dimethylaminopropyl)-3-ethylcarbodiimide (0.25 g, 1.2
mmol) in dimethylformamide (40 mL)as described in
Preparation 6 gave 0.52 g (65~) of the desired product as
a tan foam: 1H-NMR is consistent with structure; MS (FD)
728.4 (M+) ; Anal. Calc'd for C~~H9~F3N606: C, 60.98; H,
6.50; N, 11.53. Found: C, 61.18; H, 6.35; N, 11.44.
Reaction of the product of Preparation 102 (0.36 g, 0.49
mmol) and trifluoroacetic acid (4 mL) in dichloromethane
(12 mL) as described in Example 1 gave 0.3 g (88~) of the
desired mixture of isomers. Resolution of the
SUBSTITUTE SHEET (RULE 26)
Examples 46 and 47


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diastereomers (4 g, 3.6 mmol) by HPLC (Kromsil CHI-DMP
chiral stationary phase, 3A alcohol/dimethylethylamine
/heptane eluant) provided provided 0.6 (16 ~) of isomer
1 and 0.5 mg (12 ~) of isomer 2, both isolated as white
solids after formation of their respective hydrochloride
salts as described in Example 7:
Example 46. (isomer 1). 1H-NMR is consistent with
structure; tR = 6.9 min; MS (ion spray) 637.4 (M+1); Anal.
Calc'd for C3~HqqN6Oq~2.5HC1: C, 61.05; H, 6.44; N, 11.54.
Found: C, 60.89; H, 6.53; N, 11.25.
Example 47. (isomer 2) 1H-NMR is consistent with
structure; tR = 9.2 min; MS (ion spray) 637.4 (M+1); Anal.
Calc'd for C3~HqqN6Oq~2.6HC1: C, 60.75; H, 6.42; N, 11.49.
Found: C, 60.67; H, 6.63; N, 11.18.
Preparation 103
-~ F
~'O
Reaction of 3-fluorophenylacetic acid (15.0 g, 97.0 mmol)
and p-toluenesulfonic acid (3.0 g, 16 mmol) in absolute
ethanol as described in Preparation 1 gave 16.5 g (94~)
of the desired product as a colorless oil: 1H-NMR is
consistent with structure; MS (FD) 182 (M+); Anal. Calc'd
for C1aH11F~z: C, 65.92; H, 6.09. Found: C, 64.94; H,
5.99.
Preparation 104
~C
SUBSTITUTE SHEET (RULE 26)


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Reaction of the product of Preparation 103 (15.0 g, g2
mmol), N-bromosuccinimide (15.0 g, 84.5 mmol) and 48~ HBr
(4 drops) in carbon tetrachloride (80 mL) as described in
Preparation 2 gave 19.2 g (90~) of the desired product as
a colorless oil: 1H-NMR is consistent with structure; MS
(FD) 259, 261 (M+); Anal. Calc'd for CloHioBrF02: C, 46.00;
H, 3.86. Found: C, 45.71; H, 3.90.
Preparation 105
o-
N
~O
Reaction of the product of Preparation 104 (15.0 g, 58.0
mmol), 4-nitroimidazole (7.8 g, 63.8 mmol) and sodium
hydride ((2.8 g, 63.8 mmol) in dimethylformamide (200 mL)
as in desribed in Preparation 3 gave 11.13 g (65g) of the
desired product as a yellow oil: 1H-NMR is consistent
with structure; MS (FD) 293 (M+); Anal. Calc'd for
C13H12FN3~4: C, 53.24; H, 4.12; N, 14.33. Found: C, 53.12;
H, 4.22; N, 14.47.
SUBSTITUTE SHEET (RULE 26)


CA 02340344 2001-02-12
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°/
Reaction of the product of Preparation 105 (1.7 g, 5.g
mmol) with 10~ palladium on carbon (0.7 g) in
tetrahydrofuran (40 mL) under a hydrogen atmosphere
followed by coupling with the product of Preparation 1d
(2.2 g, 5.8 mmol), :1-hydroxybenzotriazole (0.86 g, 6.4
mmol) and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide
(1.3 g, 6.4 mmol) as described in Preparation 4 gave
2.05 g (60~) of the desired product as a yellow foam: 1H-
NMR is consistent with structure; MS (FD) 625 (M+); Anal.
Calc' d for C32H9oFN50, : C, 61. 43 ; H, 6 . 44; N, 11 . 19 .
Found: C, 61.28; H, 6.64; N, 11.32.
Preparation 106
N, .O
'O
O
N'
O
' ~F
O
Reaction of the product of Preparation 104 (0.12 g, 3.2
mmol) and lithium hydroxide (0.09 g, 3.84 mmol) in
SUBSTITUTE SHEET (RULE 2B)
Preparation 106


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dioxane (40 mL) and water (20 mL) as described in
Preparation 5 gave 1.91 g (1000 of the desired product
as a tan foam: 1H-NMR is consistent with structure; MS
(FD) 598 (M+) ; Anal. Calc'd for C3pH36FN5O~: C, 60.29; H,
6.07; N, 11.72. Found: C, 60.21; H, 6.41; N, 11.06.
Preparation 107
N N' _O
i. _O ~ O '~In
O
Reaction of the product of Preparation 106 (0.7 g, 1.2
mmol), 4-methylpiperidine (0.14 mL, 1.2 mmol), 1-
hydroxybenzotriazole (0.18 g, 1.3 mmol) and 1-(3-
dimethylaminopropyl)-3-ethylcarbodiimide (0.27 g, 1.3
mmol) in dimethylformamide (100 mL) as described in
Preparation 6 gave 0.52 g (64~) of the desired product
as a white solid: 1H-NMR is consistent with structure; MS
(FD) 678 (M+) ; Anal. Calc'd for C36Hq~FN6O6: C, 63 .70; H,
6.98; N, 12.38. Found: C, 63.62; H, 7.10; N, 12.31.
SUBSTITUTE SHEET (RULE 26)


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-222-
N N
~O
O
O N
~N
/~ NJ
~N
F
O
Preparation 108
2HCI
Reaction of the product of Preparation 107 (0.51 g, 0.75
mmol) and trifluoroacetic acid (2 mL) in dichloromethane
(6 mL) as described in Example 1 gave 0.24 g (49~) of the
desired mixture of isomers as a yellow solid: 1H-NMR is
consistent with structure; MS (FD) 578 (M+); Anal. Calc'd
for C31H39FN6Oq~2.7HC1: C, 54.99; H, 6.21; N, 12.41. Found:
C, 54.97; H, 6.23; N, 12.40.
O
Reaction of the product of Preparation 106 (0.7 g, 1.2
mmol), pyrrolidine (0.1 mL, 1.2 mmol), 1-
hydroxybenzotriazole (0.18 g, 1.3 mmol) and 1-(3-
dimethylaminopropyl)-3-ethylcarbodiimide (0.27 g, 1.3
mmol) in dimethylformamide (40 mL) as desribed in
Preparation 6 gave 0.54 g (69~) of the desired product
SUBSTITUTE SHEET (RULE 26)
Example 48


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as a yellow foam: 1H-NMR is consistent with structure; MS
(FD) 650 (M+) ; Anal. Calc'd for C3gHq3FN6O6~0.2CHC13: C,
60.89; H, 6.45; N, 12.46. Found: C, 60.91; H, 6.39; N,
12.36.
Example 49
N N
~O
/ O
O N
~~N
J
~N N
F
0
2HC1
Reaction of the product of Preparation 108 (0.4 g, 0.6
mmol) and trifluoroacetic acid {2 mL) in dichloromethane
(6 mL) as desribed in Example 1 gave 0.3 g (79~) of the
desired mixture of isomers as a yellow solid: 1H-NMR is
consistent with structure; MS (FD) 550 (M+); Anal. Calc'd
for CzgH35FN6Oq~2.2HC1: C, 55.21; H, 5.94; N, 13.32. Found:
C, 55.07; H, 5.91; N, 12.53.
Preparation 109
O
p F
Reaction of 2-fluorophenylacetic acid (15.0 g, 97.0 mmol)
and p-toluenesulfonic acid (2.8 g, 14.5 mmol) in absolute
ethanol (100 mL) as described in Preparation 1 gave 17.0
g (96~) of the desired product as a colorless oil: 1H-NMR
is consistent with structure; MS (FD) 182 (M+).
SUBSTITUTE SKEET (RULE 26)


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Preparation 110
lw /
Br
Reaction of the product of Preparation 109 (15.0 g, 82
mmol), N-bromosuccinimde (15.0 g, 84.5 mmol) and 48~ HBr
(3 drops) in carbon tetrachloride (80 mL) as described in
Preparation 2 gave 21 g (9$~) of the desired product as a
colorless oil: 1H-NMR is consistent with structure; MS
(FD) 260 (M+) .
Preparation 111
o_
i
o - N'
~~.- O
Reaction of the product of Preparation 110 (15.0 g, 5g
mmol), 4-nitroimidazole (7.8 g, 63.8 mmol) and sodium
hydride (2.8 g, 63.8 mmol) in dimethylformamide (200 mL)
as described in Preparation 3 gave 11.36 g (67~) of the
desired product as a white solid: 1H-NMR is consistent
with structure; MS (FD) 293.1 (M+); Anal. Calc'd for
C13H1zFN304: C, 53.24; H, 4.12; N, 14.33. Found: C, 53.54;
H, 4.18; N, 14.11.
SUBSTITUTE SHEET (RULE 26)


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Pr- eparation 112
N N O
_ ~~O
O O
O N
_',' N
J
-O N F
p
o /
Reaction of the product of Preparation 111 (1.7 g, 5,g
mmol) with 10~ palladium on carbon (0.7 g) in
tetrahydrofuran (50 mL) under a hydrogen atmosphere
followed by coupling with the product of Preparation 1d
(2.2 g, 5.8 mmol), 1-hydroxybenzotriazole (0.86 g, 6.4
mmol) and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide
(1.3 g, 6.4 mmol) as described in Preparation 4 gave 2.4
g (67~) of the desired product as a tan foam: 'H-NMR is
consistent with structure; MS (FD) 625 (M+); Anal. Calc~d
for C32HqaFN50~: C, 61.43; H, 6.44; N, 11.19. Found: C,
61.51; H, 6.50; N, 11.34.
Preparation 113
N N O
O O
O N O
~~N
J
O N F
O
Reaction of the product of Preparation 112 (2.35 g, 3~g
mmol) and lithium hydroxide (0.1 g, 4.6 mmol) in dioxane
SUBSTITUTE SHEET (RULE 26)


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(40 mL) and water (20 mL) as desribed in Preparation 5
gave 2.27 g (1000 of the desired product as a tan foam:
1H-NMR is consistent with structure; MS (FD) 598 (M+);
Anal. Calc'd for C3oH3sFN50~: C, 60.29; H, 6.07; H, 11.72.
Found: C, 60.08; H, 6.28; N, 11.42.
Preparation 114
O
N-- F
O
Reaction of the product of Preparation 113 (0.7 g, 1.2
mmol), 4-methylpiperidine (0.14 mL, 1.2 mmol), 1-
hydroxybenzotriazole (0.18 g, 1.3 mmol) and 1-(3-
dimethylaminopropyl)-3-ethylcarbodiimide (0.27 g, 1.3
mmol) in dimethylformamide (100 mL) as described in
Preparation 6 gave 0.56 g (69~) of the desired product as
a tan foam: 1H-NMR is consistent with structure; MS (FD)
678.2 (M+) ; Anal. Calc'd for C36H4~FN6O6: C, 63 .70; H,
6.98; N, 12.38. Found: C, 63.44; H, 7.05; N, 12.10.
Example 50
N N
'O
/ ~ O
O N
C /N 2HC1
/''~ J
--( ,N N F
O
SUBSTITUTE SHEET (RULE 28)


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Reaction of the product of Preparation 114 (0.53 g, 0.7g
mmol) and trifluoroacetic acid (4 mL) in dichloromethane
(12 mL) as described in Example 1 gave 0.38 g (75~) of
the desired mixture of isomers as a yellow solid: 1H-NMR
is consistent with structure; MS (FD) 578 (M+); Anal,
Calc'd for C31H3gFN6Qq~2.2HC1: C, 56.51; H, 6.30; N, 12.75.
Found: C, 56.45; H, 6.10; N, 12.43.
O
Reaction of the product of Preparation 113 (0.7 g, 1.2
mmol), pyrrolidine (0.1 mL, 1.2 mmol), 1-
hydroxybenzotriazole (0.18 g, 1.3 mmol) and 1-(3-
dimethylaminopropyl)-3-ethylcarbodiimide (0.27 g, 1.3
mmol) in dimethylformamide (100 mL) as described in
Preparation 6 gave 0.6 g (77~) of the desired product as
a tan foam. 1H-NMR is consistent with structure; MS (FD)
650 (M+) ; Anal. Calc'd for C39H43FN6O6: C, 62.75; H, 6.66;
N, 12.91. Found: C, 62.53; H, 6.58; N, 12.71.
SUBSTITUTE SHEET (RULE 26)
Preparation 115


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Example 51
O N N
/ O
O N
/N 2HC1
J
~N N F
i
O
Reaction of the product of Preparation 115 (0.46 g, 0.7
mmol) and trifluoroacetic acid (4 mL) indichloromethane
(12 mL) as described in Example 1 gave 0.44 g (1000 of
the desired mixture of isomers as a white foam: 1H-NMR is
consistent with structure. MS (high res) calc'd for
C29H36FN6~4 : 551. 2782 , Found: C, 551. 2779 . Anal . Calc' d
for C29H35FN60g'2HC1: C, 55.86; H, 5.98; N, 13.48. Found:
C, 56.09; H, 5.91; N, 13.44.
Preparation 1i6
~3
Reaction of 3-trifluoromethylphenylacetic acid (15.0 g,
73.4 mmol) and p-toluenesulfonic acid (3 g, 15.6 mmol) in
absolute ethanol (200 mL) as described in Preparation 1
gave 15.6 g (93~) of the desired product as a colorless
oil: 1H-NMR is consistent with structure; MS (FD) 232
(M+); Anal. Calc'd for CllHiiFs02: C, 56.90; H, 4.77.
Found: C, 56.93; H, 4.65.
SUBSTITUTE SHEET (RULE 26)


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3
~C
Reaction of the product of Preparation 116 (10.0 g, 44.0
mmol), N-bromosuccinimide (8.0 g, 45.3 mmol) and 48~ HHr
(4 drops) in carbon tetrachloride (70 mL), as desribed in
Preparation 2 gave 11.2 g (82~) of the desired product as
a colorless oil: ''H-NMR is consistent with structure; MS
(FD) 264 (M+) ; Anal. Calc'd for Cl~HIOBrF302: C, 42.47; H,
3.24. Found: C, 42.37; H, 3.26.
Preparation 118
0
O=N:
N
C F~
O
r
Reaction of the product of Preparation 117 (11.2 g, 36.0
mmol), 4-nitroimidazole (4.9 g, 43.2 mmol) and sodium
hydride (1.7 g, 43.2 mmol) in dimethylformamide (180 mL)
as described in Preparation 3 gave 6.22 g (50~) of the
desired product as a yellow oil: 1H-NMR is consistent with
structure; MS (FD) 343.1 (M+); Anal. Calc'd for
C14H1zF3N3~4: C, 48.99; H, 3.52; N, 12.24. Found: C, 48.74;
H, 3.63; N, 12.06.
SUBSTITUTE SHEET (RULE 26)
Preparation 117


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Preparation 119
N N O
~~O
/ /~ O
O
~C
3
Reaction of the product of Preparation 118 (2.0 g, 5.8
mmol) with 10$ palladium on carbon (0.6 g) in
tetrahydrofuran (80 mL) under an atmosphere of hydrogen
followed by coupling with the product of Preparation 1d
(2.2 g, 5.8 mmol), 1-hydroxybenzotriazole (0.86 g, 6.4
mmol) and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide
(1.3 g, 6.4 mmol) as described in Preparation 4 gave
1.82 g (47~) of the desired product as a tan foam: ~~H-NMR
is consistent with structure; MS (FD) 675.4 (M+); Anal.
Calc'd for C33H90F3N5O7: C, 58.66; H, 5.97; N, 10.36.
Found: C, 58.67; H, 5.87; N, 10.51.
Preparation 120
N N O
~O
I~ o
O N
~N
NJ
0
~CF3
Reaction of the product of Preparation 120 (1.67 g, 2.5
mmol) and lithium hydroxide (0.07 g, 2.8 mmol) in dioxane
(40 mL) and water (20 mL) as described in Preparation 5
SUBSTITUTE SHEET (RULE 26)


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gave 1.60 g (99~) of the desired product as a yellow
foam: 1H-NMR is consistent with structure; MS (FD)
648 (M+) ; Anal. Calc'd for C31H36F3N5~7: C, 57.49; H, 5.60;
N, 10.81. Found: C, 57.52; H, 5.62; N, 10.75.
Preparation 121
N N O
'O
..~.. O O
-C'r
o ll ~Y"~a
Reaction of the product of Preparation 120 (0.6 g, 0.93
hydroxybenzotriazole (0.13 g, 1.02 mmol) and 1-(3-mmol),
4-methylpiperidine (0.11 mL, 0.93 mmol), 1-
dimethylaminopropyl)-3-ethylcarbodiimide (0.12 g, 1.02
mmol)in dimethylformamide (40 mL) as described in
Preparation 6 gave 0.55 g (81~) of the desired product as
a tan foam: 1H-NMR is consistent with structure; MS (FD)
728.9 (M+) ; Anal. Calc'd for C3~Hq~F3N6O6: C, 60.98; H,
6.50; N, 11.53. Found: C, 60.81; H, 6.57; N, 11.69.
Example 52
O N N
/ O
O
~N
~ NJ
--( .N
~.J ~ ~ \ cF,
0
2HC1
SUBSTITUTE SHEET (RULE 26)


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-232-
Reaction of the product of Preparation 121 (0.5 g, 0.68
mmol) and trifluoroacetic acid (2 mL) in dichloromethane
(6 mL) as described in Example 1 gave 0.33 g (69~) of the
desired mixture of isomers as a yellow solid: 1H-NMR is
consistent with structure; MS (ion spray) 628.8 (M+1);
Anal. Calc'd for C3zH39F3N6Oq~2.3HC1: C, 53.94; H, 5.84; N,
11.79. Found: C, 53.89; H, 5.92; N, 11.65.
Preparation 122
3
Reaction of 3-trifluoromethylphenylacetic acid (15.0 g,
73.4 mmol) and p-toluenesulfonic acid (2.8 g, 14.5
mmoI)in absolute ethanol (200 mL) as described in
Preparation 1 gave 16.11 g (94~) of the desired product
as a colorless oil: 1H-NMR is consistent with structure;
MS (FD) 232 (M+) . Anal. Calc' d for C11H11F30z : C, 56. 90;
H, 4.77. Found: C, 56.64; H, 4.90.
Preparation 123
C 3
Reaction of the product of Preparation 122 (15.0 g, 65
mmol), N-bromosuccinimide (11.9 g, 67.0 mmol) and 48~ HBr
(4 drops) in carbon tetrachloride (80 mL) as described in
Preparation 2 gave 17,1 g (85~) of the desired product as
SUBSTITUTE SHEET (RULE 26)


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a colorless oil: 1H-NMR is consistent with structure; MS
(FD) 311, 313 (M+),
Preparation 124
o-
o=N,
N C F~
~O
Reaction of the product of Preparation 123 (15.0 g, 48.0
mmol), 4-nitroimidazole (6.0 g, 52.8 mmol) and sodium
hydride (2.1 g, 52.8 mmol) in dimethylformamide (200 mL)
as described in Preparation 3 gave 12.25 g (74~) of the
desired product as a yellow oil: 1H-NMR is consistent with
structure; MS (FD) 343 (M+); Anal. Caic'd for C14Hi2F3N304:
C, 48.99; H, 3.52; N, 12.24. Found: C, 49.10; H, 3.58;
H, 12 . 22 .
Preparation 125
N N O
_O
O N O O
''N
J
~O N CF3
O
'''
Reaction of the product of Preparation 124 (2.0 g, 5.8
mmol) with 10~ palladium on carbon (1.0 g)in
tetrahydrofuran (60 mL) under a hydrogen atmosphere
follwed by coupling with the product of Preparation ld
(2.2 g, 5.8 mmol), 1-hydroxybenzotriazole (0.86 g, 6.4
SUBSTITUTE SHEET (RULE 2B)


CA 02340344 2001-02-12
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mmol) and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide
(1.3 g, 6.4 mmol) as described in Preparation 4 gave 3.16
g (93~) of the desired product as a tan foam: 1H-NMR is
consistent with structure; MS (FD) 675.4 (M+); Anal.
Calc'd for C33H40F3N5~7: C, 58.66; H, 5.97; N, 10.36.
Found: C, 58.81; H, 6.04; N, 10.12.
Preparation 126
N N O
~O /\
O N O O
1'' N
J
O N CF3
O
Reaction of the product of Preparation 125 (2.78 g, 4.1
mmol) with lithium hydroxide (0.12 g, 4.9 mmoI) in
dioxane (40 mL) and water (20 mL) as described in
Preparation 5 gave 2.6 g (98~) of the desired product as
a yellow foam: 1H-NMR is consistent with structure; MS(FD)
648.2 (M+) ; Anal. C"alc'd for C31H3sF3NsO~: C, 57.49; H,
5.60; N, 10.81. Found: C, 58.06; H, 6.14; N, 10.27.
Preparation 127
N N, _O
_ O ~~.~(,
/ ~ O O
O
~~N
~.-~N N C F3
O
Reaction of the product of Preparation 126 (0.7 g, 1.1
mmol), 4-methylpiperidine (0.13 mL, 1.1 mmol), 1-
SUBSTITUTE SHEET (RULE 26)


CA 02340344 2001-02-12
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-235-
hydroxybenzotriazole (0.17 g, 2.2 mmol) and 1-(3-
dimethylaminopropyl)-3-ethylcarbodiimide (0.25 g, 1.2
mmol)in dimethylformamide (30 mL) as described in
Preparation 6 gave 0.32 g (40~) of the desired product
as a tan foam: 1H--NMR is consistent with structure; MS
(FD) 728 (M+) ; Anal. Calc'd for C3~Hq~F3N6O6: C, 60.98; H,
6.50; N, 11.53. Found: C, 60.76; H, 6.59; N, 12.36.
Example 53
N N
~O
O
O N
2HC1
3
Reaction of the product of Preparation 127 (0.3 g, 0.41
mmol) and trifluoroacetic acid (2 mL) in dichloromethane
(6 mL) as described in Example 1 gave 0.28 g (97~) of
the desired mixture of isomers as a white solid: 1H-NMR is
consistent with structure; MS (FD) 628 (M+); Anal. Calc'd
for C32H39F3N6Oq~2.2HC1: C, 54.22; H, 5.86; N, 11.85.
Found: C, 54.33; H, 5.84; N, 11.56.
Preparation 12$
N N O
~~O
/ ~ O O
O
~'N
CN N CFs
O
SUBSTITUTE SHEET (RULE 26)


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Reaction of the product of Preparation 126 (0.5 g, 0.77
mmol), pyrrolidine (0.07 mL, 0.77 mmol), 1-
hydroxybenzotriazole (0.12 g, 0.85 mmol) and 1-(3-
dimethylaminopropyl)-3-ethylcarbodiimide (0.18 g, 0.85
mmol) in dimethylformamide (30 mL) as described in
Preparation 6 gave 0.2 g (37~) of the desired product as
a tan solid: 1H-NMR is consistent with structure; MS (FD)
700 (M+) ; Anal. Calc'd for C35H43F3N606~0.4H20: C, 59.38; H,
6.24; N, 11.87. Found: C, 59.17; H, 6.24; N, 11.87.
Example 54
N N
~O
O
O N
~N 2HC1
J
~N N CF3
O
Reaction of the product of Preparation 128 (0.2 g, 0.29
mmol) and trifluoroacetic acid (2 mL) in dichloromethane
(6 mL) as described in Example 1 gave 0.18 g (1000 of
the desired mixture of isomers as a white solid:lH-NMR is
consistent with structure; MS (FD) 600 (M+).
Preparation 129
N N O--?(
~~O
O
O
N
j N C F3
O
Reaction of the product of preparation 126 (0.75 g, 1.2
mmol), dimethylamine hydrochloride (0.1 g, 1.2 mmol),
SUBSTITUTE SHEET (RULE 26)


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triethylamine (0.19 g, 1.3 mmol), 1-hydroxybenzotriazole
(0.18 g, 1.3 mmol) and 1-(3-dimethylaminopropyl)-3-
ethylcarbodiimide (0.27 g, 1.3 mmol) in dimethylformamide
(40 mL) as described in Preparation 6 gave 0.49 g (60~)
of the desired product as a tan foam: 1H-NMR is
consistent with structure; MS (FD) 675 (M+); Anal. Calc'd
for C33HQ1F3N6O6: C, 58.75; H, 6.13; N, 12.46. Found: C,
58.69; H, 6.12; N, 12.28.
Example 55
N N
-O
O
O
~N~
/N CF3
O
2HC1
Reaction of the product of Preparation 129 (0.42 g, 0.62
mmol) and trifluoroacetic acid (2 mL) in dichloromethane
(6 mL) as described in Example 1 gave 0.3 g (75$) of the
desired mixture of isomers as a yellow solid: 1H-NMR is
consistent with structure; MS (FD) 574 (M+); Anal. Calc'd
for CZ8H33F3N6O4'2.8 HC1: C, 48.70; H,, 5.33; N, 12.42.
Found: C, 49.84; H, 5.27: N, 12.09.
SUBSTITUTE SHEET (RULE 28)


CA 02340344 2001-02-12
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-238-
Preparation 130
N N O-7(
O
O
Reaction of the product of Preparation 126 (0.5 g, 0.77
mmol), 4-(4-fluorobenzoyl)piperidine hydrochloride (0.19
g. 0.77 mmol), triethylamine (0.12 mL, 0.85 mmol), 1-
hydroxybenzotriazole (0.12 g, 0.85 mmol) and 1-(3-
dimethylaminopropyl)-3-ethylcarbod.iimide (0.18 g, 0.85
mmol) in dimethylformamide (40 mL) as described in
Preparation 6 gave 0.45 g (69~) of the desired product as
a yellow foam: 1H-NMR is consistent with structure; MS
(FD) 836.8 (M+) ; Anal. Calc'd for Cq3H4eF41V6~7-0.4H20: C,
61.19; H, 5.83; N, 9.96. Found: C, 60.92; H, 5.56; N,
10.09.
Example 46
N N
~O
/ O
O
'N 2NCI
J
F ~ ~ N N Ct=3
O O
Reaction of the prodcut of Preparation 130 {0.4 g, 0.48
mmol) and trifluoroacetic acid (2 mL) in dichloromethane
(6 mL) as described in Example 2 gave 0.26 g (67~) of the
SUBSTITUTE SHEET (RULE 26)


CA 02340344 2001-02-12
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-239-
desired mixture of isomers as a white solid. -H-NMR is
consistent with structure; MS (FD) 736.7 (M+); Anal.
Calc'd for C38HQOF4N605~2.1 HC1: C, 56.12; H, 5.22; N,
10.33. Found: C, 56.p8; H, 5.46; N, 10.38.
Preparation 131
0
i
p~N~
N
~-O
0
Reaction of alpha-bromocyclohexylacetic acid (5.0 g, 21.0
mmol), 4-nitroimidiazole (2.6 g, 23.1 mmol) and sodium
hydride (0.93 g, 23.1 mmol) in dimethylformamide (45 mL)
as described in Preparation 3 gave 1.9 g (34~) of the
desired product as a clear oil: 1H-NMR is consistent with
structure; MS (ion spray) 268 (M+1); Anal. Calc'd for
C12H1~N3~4: C, 53.92; H, 6.41; N, 15.72. Found: C, 53.63;
H, 6.33; N, 15.77.
Preparation 132
"O
O
Reaction of the product of Preparation 131 (1.4 g, 5,2
mmol) with 10~ palladium on carbon (0.8 g) in
SUBSTITUTE SHEET (RULE 26)


CA 02340344 2001-02-12
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-240-
tetrahydrofuran (60 mL) under a hydrogen atmosphere
followed by coupling with the product of Preparation ld
(2.0 g, 5.2 mmol), 1-hydroxybenzotriazole (0.8 g, 5.7
mmol) and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide
(1.2 g, 5.7 mmol) as described in Preparation 4 gave
2.09 g (65~) of the desired product as a tan foam. 1H-NMR
is consistent with structure; MS (ion spray) 600.4 (M+1);
Anal. Calc'd for Cj1H45N5O7: C, 62.08; H, 7.56; N, 11.68.
Found: C, 62.04; H, 7.53; N, 11.74.
Preparation 133
'O
Reaction of the product of Preparation 132 (2.0 g, 3.3
mmol) with lithium hydroxide (0.1 g, 4.0 mmol) in dioxane
(50 mL) and water (25 mL) as described in Preparation 5
gave 1.9 g (99~) of the desired product as a tan foam: 1H-
NMR is consistent with structure; MS (ion spray) 586.4
(M+1 ) ; Anal . Calc' d for C3oH43N50~ : C , 61. 52 ; H, 7 . 4 0 ; N,
11.96. Found: C, 61.41; H, 7.42; N, 11.82.
SUBSTITUTE SHEET (RULE 26)


CA 02340344 2001-02-12
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-241-
Preparation 134
N N O--7(
\. ~ ~ ~O
O O
O N
C %N
N N.J
O
Reaction of the product of Preparation 133 (0.8 g, 1.4
mmol), 4-methylpipe:ridine (0.17 mL, 1.4 mmol), 1-
hydroxybenzotriazole (0.21 g, 1.54 mmol) and 1-(3-
dimethylaminopropyl)-3-ethylcarbodiimide (0.32 g, 1.54
mmol) in dimethylformamide (30 mL) as described in
Preparation 6 gave 0.92 g (99~) of the desired product as
a tan foam: 1H-NMR is consistent with structure; MS (ion
spray) 667.5 (M+1); Anal. Calc'd for C36HSQN6O6: C, 64.84;
H, 8.16; N, 12.60. Found: C, 64.55; H, 7.73; N, 12.26.
Example 57
\ ~N
O
2HC1
Reaction of the product of Preparation 134 (0.7 g, 1.0
mmol) and trifluoroacetic acid (2 mL) in dichloromethane
(6 mL) as described in Example 1 gave 0.43 g (64~) of the
desired mixture of isamers as a tan solid: 1H-NMR is
SUBSTITUTE SHEET (RULE 2B)


CA 02340344 2001-02-12
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-242-
consistent vuith structure; MS (ion spray) 567.6 (M+1);
Anal. Calc'd for C31Hg6N6Oq~2HCl: C, 58.21; H, 7.56; N,
13.14. Found: C, 58.36; H, 7.33; N, 13.19.
Preparation 135
N N O
I~ o
O~N O
~~N
N
O
Reaction of the product of Preparation 133 (0.8 g, 1.4
mmol), dimethylamine hydrochloride (0.22 g, 1.4 mmol),
triethylamine (0.22 mL, 1.54 mmol), 1-
hydroxybenzotriazo:le (0.21 g, 1.54 mmol) and 1-(3-
dimethylaminopropyl)-3-ethylcarbodiimide (0.32 g, 1.54
mmol) in dimethylformamide (30 mL) as described in
Preparation 6 gave 0.86 g (1000 of the desired product
as a tan foam: 1H-NMR is consistent with structure; MS
{ ion spray) 613 . 4 (M+1 ) ; Anal . Calc' d for C32HQgN6O6 : C,
62.72; H, 7.90; N, 13.72. Found: C, 62.44; H, 7.64; N,
13.57.
SUBSTITUTE SHEET (RULE 26)


CA 02340344 2001-02-12
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Example 58
n0 N~N
OO
O
~N 2HCI
N''J
O
Reaction of the product of Preparation 135 (0.7 g, 1.0
mmol) and trifluoroacetic acid (2 mL) in dichloromethane
(6 mL) as described in Example 1 gave 0.43 g (64$) of the
desired mixture of isomers as a tan solid: 1H-NMR is
consistent with structure; MS (ion spray) 567.6 (M+1);
Anal. Calc'd for C31Hq6N6Oq~2HCl: C, 58.21; H, 7.56; N,
13.14. Found: C, 58.36; H, 7.33; N, 13.19.
Preparation 136
U,
.>
To a suspension of 2-naphthyl acetic acid (49.37 g, 265.0
mmol) in carbon tetrachloride (55 mL) was added and
thionyl chloride (80 mL). The mixture was heated to
reflux for 20 minutes then cooled to ambient temperature.
Carbon tetrachloride (125 mL), N-bromosuccinimide (56.60
g, 318.0 mmol) and hydrobromic acid (48~ aq., 0.5 mL)
were added. The mixture was heated to reflux for 30 min,
SUBSTITUTE SHEET (RULE 26)
O
N


CA 02340344 2001-02-12
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-244-
cooled to ambient temperature, filtered, and
concentrated. The resulting material was dissolved in
dichloromethane (200 mL) and excess ethanol (100 mL) was
added dropwise. After 1 h, the reaction was concentrated
S and the resulting crude material was purified by flash
chromatography(si:Lica gel, 30~ ethyl acetate/hexane) to
yield a tan solid. This crude material was dissolved
dimethylformamide (200 mL) and 4-nitroimidazole (29.78 g,
263.5 mmol) and potassium carbonate (72.70 g, 526.8 mmol)
were added. After 16 h, the reaction was concentrated
to 100 mL. Ethyl acetate and water were added and the
mixture washed with sodium bicarbonate and brine. The
organic layer was dried over sodium sulfate and
concentrated . The crude material was was purified by
flash chromatography (silica, 30~ ethyl acetate/hexane)
to yield 40.2 g (47~) of the desired product as a brown
foam: 1H NMR (300 MHz, CDC13) - consistent with
structure; Anal. calcd. for C1~H15N309; 62.76 C, 4.65 H,
12.92 N; found 60.54 C, 4.35 H, 12.04 N; ISMS (M+) - 326.
Preparation 137
,>
SUBSTITUTE SHEET (RULE 26)


CA 02340344 2001-02-12
WO 00/10565 PCT/US991035Z5
-245-
Reaction of the product of Preparation 136 (4.80 g, 14.77
mmol) with 5~ palladium on carbon (2.5 g) in
tetrahydrofuran (100 mL) under a hydrogen atmosphere
followed by coupling with the product of Preparation 1d
(5.61 g, 14.77 mmol), EDCI (2.79 g, 16.25 mmol), 1-
hydroxybenzotriazole (2.00 g, 14.77 mmol), and N-
methylmorpholine (1.6 mL, 14.77 mmol) as described in
Preparation 4 gave (6.04 g, 62~) of the desired product
as a light orange foam: 1H NMR (300 MHz, CDC13) -
consistent with structure; Anal. calcd. for C36H43N5~7;
65.74 C, 6.59 H, 10.65 N; found 64.02 C, 6.09 H, 10.13 N;
ISMS (M+) - 658.
Preparation 138
A solution of lithium hydroxide (0.38 g, 9.16 mmol) in
water (50 mL) was added to a solution of the product of
Preparation 137 (6.04 g, 9.16 mmol) in tetrahydrofuran
(100 mL). After 30 min, water was added and the mixture
washed with diethyl ether. The aqueous layer was
adjusted to pH = 3.0 with sodium bisulfate, saturated
with sodium chloride, and washed with ethyl acetate. The
combined organic extracts were dried over sodium sulfate,
and concentrated. T~o the resulting crude material
stirring at room temperature in dimethylformamide (50 mL)
was added 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide
SUBSTITUTE SHEET (RULE 26)


CA 02340344 2001-02-12
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-246-
(2.08 g, 10.01 mmol.), 1-hydroxybenzotriazole (1.24 g,
9.16 mmol) and 4-methyipiperidine (1.1 mL, 9.16 mmol).
After 18 h, the reaction was quenched with saturated
bicarbonate, and washed with ethyl acetate. The combined
organic extracts were washed with brine, dried over
sodium sulfate, and concentrated. The crude material was
purified by flash chromatography (silica gel, 5~
methanol/dichloromethane) to yield 4.9 g (75 ~s) of the
desired product as a pale yellow foam: 1H NMR (300 MHz,
CDC13) - consistent with structure; Anal. calcd. for
C40H50N6~6% 67.58 C, 7.09 H, 11.82 N; found 65. 60 C, 7.09
H, 11.50 N; ISMS (M~+) - 711.
Examples 59 aad 60
CIH
CIH
N N
To a solution of of the product of Preparation 138 (4.90
g, 6.89 mmol) stirring at room temperature in
dichloromethane (40 mL) and anisole (1.0 mL) was added to
triflouroacetic acid (10 mL). After 3 hours, the
reaction was quenched with saturated sodium bicarbonate
and extracted with ethyl acetate. The combined organic
extracts were washed with brine, dried over sodium
sulfate and concentrated. The resulting crude material
was purified by flash chromatography (silica gel, 5~
methanol/dichloromethane) to give the product as a
SUBSTITUTE SHEET (RULE 26)


CA 02340344 2001-02-12
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mixture of diastereomers. This material was resolved by
HPLC (Kromsil CHI-~DMP chiral stationary phase, 3A
alcohol/ dimethylethylamine/heptane eluant) to provide
the free amine of the desired products. The individual
diastereomers were dissolved in ethyl acetate and treated
with a saturated solution of hydrochloric acid in diethyl
ether. The resulting precipiate was filtered to yield
the desired products (426779 - 0.64 g, 14~) (426780 -
0.43 g, 9~) as tan solids: Example 59. 1H NMR (300 MHz,
CDC13) - consistent with structure; Anal. calcd. for
C35H44N6~4C12; 61.49 C, 6.49 H, 12.29 N; found 60.28 C, 6.38
H, 11.74 N; ISMS (M+) - 611. Example 60. 1H NMR (300
MHz, CDC13) - consistent with structure; Anal. calcd. for
C35H44N6~4C12; 61.49 C, 6.49 H, 12.29 N; found 47. 81 C, 5.29
H, 9.83 N; ISMS {M+) - 611.
Preparation 139
O
N ~
N- 'O
O N O
Nw%N
O
Reaction of the product of Preparation 136 (1.318, 4.02
mmol) with 10~ palladium on carbon {0.5 g) in
tetrahydrofuran (50 mL) under a hydrogen atmosphere
followed by coupling with the product of Preparation 1j
(1.528, 4.02 mmol), 1-hydroxybenzotriazole (0.688, 4.42
mmol), and 1-(3-dimethylaminopropyl)-3-
ethylcarbodiimide(0.91g, 4.42 mmol) as described in
SUBSTITUTE SHEET (RULE 26)


CA 02340344 2001-02-12
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-248-
Preparation 4 to give g (38 of the title compound
1.06 ~)


as a tan sol id: 1H NMR DMSO, 1.22(m, 18H), 1.50(m,
(d6- d):


4H), 2.55(m, 2H), 4.26(q, J = 9.OHz, 2H), 4.37(bs, 1H),


5.75(s, 1H), 6.60(s, 1H), 7.02(bs, 1H), 7.I6(m, 3H),


7.22(m, 3H), 7.43(m., 1H),7.50(d, = 9.3Hz, 2H), 7.60(m,
J


2H), 7.97(m, 3H), 10.21(m,1H). Ion spray MS (M' +1):


656.


Preparation 140
O
N
N
/ O~ O
N
O
NON
O
Reaction of the product of Preparation 139 (1.06 g, 1.62
mmol) with lithium hydroxide 75 mg, 1.78 mmol)in dioxane
(30 mL) and water (1.5 mL) as described in Preparation 5
gave 1.01 g (100 ~) of the title compound as a golden
yellow solid: 1H NMR (d6-DMSO, d): 1.20(m, 15H),
1.50(m,4H), 2.55(m, 2H), 4.38(bs, 1H), 6.58(s, 1H),
7.02(bs, 1H), 7.17(m, 3H), 7.25(m, 3H), 7.35(m, 1H),
7.50(m, 2H), 7.58(m, 2H), 7.98(m, 3H), 8.09(m, 1H),
10.36(bs, 1H). Ion spray MS (M+ +1): 628.
SUBSTITUTE SHEET (RULE 2B)

CA 02340344 2001-02-12
s
t
DEMANDES OU BREVETS VOLUMINEUX
LA PRESENTS PART1E DE CETTE OEMANDE OU CE BREVET
COMPREND PLUS D'UN TOME.
CECt EST LE TOME ~ DE 3
NOTIE: Pour ies tomes additionels, veuiiiez cantacter le Bureau canadien des
brevets
JUMBO APP~ICAZ'IONS/PAi'ENTS
THIS SECTION OF THE APPL1CATION/PATENT CONTAINS MORE
THIS IS VOLUME ~_ OF
MOTE: For additional voiumes~please canta'ct'the Canadian Patent Office