CA2340344A1 - Growth hormone secretagogues - Google Patents

Abstract

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

Description

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DEMANDES OU BREVETS VOLUMINEUX
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THIS IS VOLUME ~_ OF J
I110TE:'For additionay voiumes-ple.ase cnntact'the Canadian Patent Office 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) 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) 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 United States Patent 5,206,235, issued April 27, 1993, describes a series of benzolactam compounds typified by the following structure.
H

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.

~ H
N~~ ~3+ CH3S03 N
I ~ ~ N~

SUBSTITUTE SHEET (RULE 26) 5 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

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) 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) /\ /~ ~~ ~~ /~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~

N ~ \
R4 N. N / NJ
I I
N NH Z , ~ /
I l N
SUBSTITUTE SHEET (RULE 28) 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) 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) N N N N N

N N N
NCl--C6alkyl NC1-C6alkylaryl N N ,N N N N
C
S O ~N ~ 0 Naryl 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~

wo oonos6s Pcrivs~ro35zs 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) 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~

N

or NH
SUBSTITUTE SHEET (RULE 2B) 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) N N N N
C> > o C , ~I
I
N
CND
N N N N
C~ C~ C~ C~
S I N I
p p~ SOZCH
N
U N~ U

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) H

~NH2 O O
HN
N
Y

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) H
N
- NHZ
/ O O
NH
Hjy N

O H
IA
The present invE~ntion still further relates to compounds of formula TB:
H
N
NHZ

NH
N
N ~ ~ CHs IB
The present invention additionally relates to compounds of formula Ia':
SUBSTITUTE SHEET (RULE 26) H
N
NHBoc NH
N
EtO
-E

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:

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) "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) 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) 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) WO 00/t0565 PCT/US99/03525 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) 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) 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 NHZ
SUBSTITUTE SHEET (RULE 26~

J is H;
G is H;
X is NH;
V is /~ /\ /
N
N N N~
I ~ or E is ~ ~ ~ OMe ~~CH3 / ~ \ / OMe \ \ ~ \
/ ~ ~ / ~ /

\ ~ ~ \
/ F ~ O~\
D is -C (0) R6, where R6 is 1-pyrrolidinyl, 1-piperidinyl, 4-methyl-1-piperidinyl, N,N-dimethyl, N ~ or N ~ ~ F
L is H or CH3;
SUBSTITUTE SHEET (RULE 26) 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) 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) WO 00/10565 PC'T/US99/03525 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) Scheme I

N
N
Br H N
Et0 V Et R
R
O
NaH, DMF O
IV
IIb' LiOH

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

R
O
II
wherein R is representative of E as previously defined, and R2R1N is R6 as previously defined.
;iUBSTITUTE SHEET (RULE 26) 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) 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) 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 OMe IIIa' LiOH
H
\ N
0 NHBoc OH
III
SUBSTITUTE SHEET (RULE 26) Scheme II, continued IIb' DCC, HOBT
H

NHBoc ~i 0 0 NH
N
Et0 R
O
Ia' 1. LiOH
2. RIRzNH VI
3. TFA
H
N

NH
N

R
O
Ia .SUBSTITUTE SHEET (RULE 26) 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) Scheme III
NOZ NOZ
r _ I~, \ ; 1. LiOH
N N
H NaH
DMF Et0 \ Ph 2.
VII NH
O
VI I I ' C OZMe 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

NH TFA NH
r r Dl N
N ~ N
Ph Ph COZMe O C02Me 0 Ib' Ib SUBSTITUTE SHEET (RULE 2B) 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) 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) 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~

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) 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) 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) WO 00/10565 PCT/US99/fl3525 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) 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) WO 00/10565 PC'F/US99/03525 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) 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) 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) WO 00/1O5b5 PCT/US99/03525 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) 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) 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) 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) 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) 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) Formulation 4 Tablets, each containing 30 mg of active ingredient, are prepared as follows:
Quantity Ingredient (mg/tablet) Active Ingredient 30 .
mg Starch 45.0 mg Microcrystalline cellulose 35 .
mg Polyvinylpyrrolidone (as 10~ solution in water) 4.0 mg Sodium carboxymethy.l starch 4 .
mg Magnesium stearate 0 .
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) 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) 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) 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) 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.
SUBSTITUTE SHEET (RULE 2B) Preparation 1 O NHBOC
CO Me 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$) as a light yellow oil: 1H NMR (300 MHz, CDC13) d 1.45 (s, 9H), 3.70 (m, 1H), 3.75 (s, 3H), 3.85 (m, 1H), 4.50 (m, 3H), 7.30 (m, 5H); MS (FD) m/e 310; Anal. calc'd for C16H23N05: C, 62.12; H, 7.49; N, 4.53. Found: C, 62.31; H, 7.49; N, 4.43.
Preparation 2 o~r~2 COZMe To a solution of a compound of Preparation 1 (5.0 g, 16 mmol) stirring in dichloromethane (25 mL) and anisole (1 mL) at 0 oC was added trifluoroacetic acid. After 4 h at room SUBSTITUTE SHEET (RULE 26) WO 00/IOSbS PCT/US99/03525 temperature, saturai:ed sodium bicarbonate solution was added and the mixture extracted with ethyl acetate. The combined organic extracts were washed with brine, dried over sodium sufate, and concentrated. The crude product was used in the next step without further purification.
Preparation 3 H
'~ O ~ N NHBoc Me0 2C O
To a solution of a compound of Preparation 2 (65.4 mmol), boc-a-aminoisobutyric acid (13.2 g, 65.4 mmol), 1-hydroxybenzotriazole~ (8.8 g, 65.4 mmol), and N,N-diisopropylethylamine (22.8 mL, 130.7 mmol) stirring in dichloromethane ( 500 mL) at 0 °C was added 1- ( 3-dimethylaminopropyl)-3-ethylcarbodiimide (12.3 g, 71.9 mmol). After 18 h, ethyl acetate and saturated ammonium chloride were added and the mixture extracted with ammonium chloride, sodium bicarbonate, and brine. The organic extracts were dried over sodium sulfate and concentated.
Purification by silica gel chromatography (25$ 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) Preparation 4 H
O~ N NHBoc 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 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$) oi= the desired product as a white solid:
1H-NMR is consistent with structure; MS (FD) 275 (M+); Anal.
Calc'd. for: C, 56.73; H, 4.73; N, 15.27. Found: C, 56.48;
H, 4.78; N, 15.08.
SU8ST1TUTE SHEET (RULE 26) Preparation 7 Bc To a suspension of 5$ 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) 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$) as a light orange foam: 1H NMR (300 MHz, CDC13) d ; MS (FD) m/e Anal, calc'd for . C, ; H,; N, . Found: C, ; H, ; N, .
Preparation 9 N~
_ N
N~ O
H N
SUBSTITUTE SHEET (RULE 2B) To a solution of a compound of Preparation 8 (2.09 g, 3.61 mmol), pyrrolidine (0.30 mL, 3.61 mmol), and 1-hydroxybenzotriazole (0.59 g, 3.97 mmol) stirring in anhydrous DMF (50 mL) at 0 °C was added 1,3-dicyclohexyl carbodiimide (0.82 ~g, 3.97 mmol). After 18 hours at room temperature, the reaction was concentrated, dissolved in dichloromethane, filtered, and concentrated. Purification by silica gel chromatography (5$ 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~

~~ 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) 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) Preparation 11 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$) of the desired product as a. clear oil. 1H-NMR is consistent with structure; MS (FD) 303 (M+); Anal. Calc'd for: C, 59.40; H, 5.65; N, 13.85. Found: C, 59.73; H, 5.71; N, 13.40.
SUBSTITUTE SHEET (RULE 26) WO 00/10565 PC'T/US99/03515 Preparation I2 N
N
H:O
p To a solution o:f a compound of Preparation 11 (4.24 g, 14 mmol) stirring in tetrahydrofuran (30 mL) and ethanol (30 mL) at room temperai~ure was added 2N NaOH (35 mL, 70 mmol) .
After 1 h, this mixture was treated with 5N HC1 until pH =
2.5. Ethyl acetate ~;30 mL) and water (30 mL) were added and the resulting solution was extracted with ethyl acetate.
The combined organic: extracts were washed with brine, dried over sodium sulfate, filtered and concentrated to give 3.8 g (98$) of the desired product as a yellow oil: 1H-NMR is consistent with stru.cture~ MS (FD) 276 (M+), SUBSTITUTE SHEET (RULE 2B) wo oon os6s pcT~smo3s2s 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$) of the desired product as a yellow oil: 1H-NMR is consistent with structure; MS (FD) 386.2 (M+).
SUBSTITUTE SHEET (RULE 26) Preparation 14 O N
NHBoc O O
HN
N
N
N
_ v MfeO 2 C .
To a slurry of 10~ Pd/C in dioxane (10 mL) was added a solution of a compound of Preparation 13 (2.4 g, 6.2 mmol) in dioxane (100 mL). The mixture was then treated with hydrogen gas (40 psi) on a Parr apparatus. After 5 h, an amount of 10$ 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) WO 00/10565 PC"T/US99/03525 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) _70-Preparation 15 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 N
Et0 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$) of the desired product as a yellow foam: 1H-NMR
is consistent with structure; MS (FD) 289.1 (M+); Anal.
Calc'd. for: C, 58.13; H, 5.23; N, 14.53. Found: C, 58.40;
H, 5.17; N 14.24.
SUBSTITUTE SHEET (RULE 26) Preparation 17 NOz N
HO
O
s Reaction of a compound of Preparation 16 (3.3 g, 12.0 mmmol ) , 2N NaOH ( 30 mL, 60 mmol ) in ethyl acetate ( 30 mL)/ethanol (30 mL) according Preparation 12 gave 2.85 g (90$) 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 SUBSTITUTE SHEET (RULE 26) 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

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$) of the desired product as a tan foam: 1H-NMR is consistent with structure;
MS (FD) 704.5 (M+) , .SUBSTITUTE SHEET (RULE 26) Example 3 H
O N
'\~~~~ 2 N
<~ N I
N
Me0 ZC
Reaction of a compound of Preparation 19 (0.23 g, 0.33 mmol) , trifluoroacet:ic acid (4.0 mL, 24 mmol) in dichloromethane (12 mL), followed by treatment with HC1/ethyl acetate solution (40 mL), according to Example 2 gave 0.17 g (77$) 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 N
Et0 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) 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 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 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) (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.

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) Example 4 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$) of the desired product as an off-white solid: 1H-NMR
is consistent with :>tructure; MS (FD) 514 (M+); Anal. Calc'd for: C, 58.35; H, 5.66; N, 16.33. Found: C, 58.25; H, 6.40;
N, 16.16.
Preparation 24 Reaction of 5-nitroindole (3.0 g, 18.5 mmol), a-bromophenylacetic acid ethylester (4.5 g, 18.5 mmol), and sodium hydride (0.8 ct, 20 mmol, 60$ dispersion in mineral oil) in N,N-dimethylfarmamide (75 mL) according to Preparation 1 gave 3.9 g (65$) of the desired product: 1H-SUBSTITUTE SHEET (RULE 26) WO 00/10565 PGT/(JS99/035Z5 NMR is consistent with structure; MS (FD) 324 M+; Anal.
Calc' d for: C, 66. E~6; H, 4.97; N, 8. 64 . Found: C, 66. 80; H, 5.11; N, 8.81, Preparation 25 N
HO
I
O
Reaction of a compound of Preparation 24 (2.0 g, 6.2 mmol) and 2N NaOH (5.0 mL, 100 mmol) in tetrahydrofuran (10 mL)/ethanol (8 mL) according to Preparation 12 gave 1.4 g (76$) 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) _78_ Preparation 26 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) _79_ 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$) of the desired product: 1H-NMR is consistent with structure;
MS (FD) 739.7 (M+) Example 5 O
IV~i2 O O
HN
Me0 2C
Reaction of a compound of Preparation 27 (0.38 g, 0.51 mmol) and trifluoroacetic acid (2 mL, 26 mmol) in dichloromethane (10 mL) according to Example 2 gave 0.125 g (38~) of the desired product: 1H-NMR is consistent with structure; MS (FD) 6:39 (M+) ; Anal . Calc' d for 1 H20: C, 65.74; H, 6.59; N, 10.65. Found: C, 65.75; H, 6.42; N, 10.98.
SUBSTITUTE SHEET (RULE 28) Preparation 28 ~"'~ N
N
HO
To a solution ~of a compound of Preparation 6 (27 g, 98 mmol) stirring in tetrahydrofuran (60 mL) and absolute ethanol (60 mL) at ambient temperature was added 2N NaOH
(250 mL, 500 mmol). After 3.5 h, the mixture was washed with diethyl ether and the organic extract subsequently washed with water. The combined aqueous extracts were acidified and the rE:sulting mixture extracted with ethyl acetate. The combined organic extracts were washed once with brine, dried over sodium sulfate, filtered, and concentrated to give' 24.2 g (75$) 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 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~

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) 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) Preparation 32 ~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 HN
MeO 2C
2.5 HC1 SUBSTITUTE SHEET (RULE 2t3) 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) 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) 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.

Example 7 ,..~~NH 2 HN
i MfeO 2 C
SUBSTITUTE SHEET (RULE 2B) _87_ 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) _88_ 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) _89_ 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$) of the desired compound as a white solid: 1H-NMR is consistent with structure; MS (FD) 590 (M+); Anal. Calc'd for: C, 56.11; H, 6.08; N, 12.66. Found: C, 56.16; H, 5.92; N, 12.56. IR
(KBr) 3163.75, 3031..15, 2952.46, 2876.38, 1745.07, 1664.94, 1530.69, 1497.79, 1453.37, 1435.81, 1197.21, 1177.62, 1094.93, 747.95, 701..04 cm 1.
SUBSTITUTE SHEET (RULE 28) WO 00/10565 PC'f/US99/03525 Preparation 40 O~
~N
O
To a solution of a compound of Preparation 28 (1.0 g, 4.0 mmol), morpholine (0.35 mL, 4.0 mmol), 1-hydroxybenzotriazolE~ hydrate (0.6 g, 4.4 mmol) stirring in N,N-dimethylformamide (50 mL) at room temperature was added 1,3-dicyclohexylcarbodiimide (0.9 g, 4.4 mmol). After 16 h, the mixture was concentrated, and the residue extracted with ethyl acetate. The combined organic extracts were filtered, washed with saturated aqueous sodium bicarbonate, water, brine, dried over sodium sulfate, filtered, and concentrated. The resulting residue was purified by silica gel chromatography (methanol/chloroform) to give 0.75 g (60$) of the desired product as a white foam: 'H-NMR is consistent with structure; MS (FD) 316 (M+).
SUBSTITUTE SHEET (RULE 26) WO 00/10565 PC'T/US99/03525 Preparation 41 H
/~ 0 N
~~~ NHBoc O O
HN
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) WO 00/10565 PC'f/US99/03525 Example 9 I 0 ~'~ NH z HN
N
~N
O

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) Preparation 42 NOz CI N
N

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$) of the desired product as a tan foam: 'H-NMR is consistent with structure; MS (FD) 314 (M+).
Preparation 43 O
NHBoc O O
c Hydrogenation of a compound of Preparation 42 (0.91 g, 2.9 mmol) in ethyl acetate (50 mL)/ethanol (50 mL), 5~ Pd/C
(0.36 g) in ethyl acetate (5 mL) followed by reaction with a compound of Preparation 4 (0.95 g, 2.5 mmol), 1-hydroxybenzotriazole hydrate (0.37 g, 2.75 mmol), and 1,3-SUBSTITUTE SHEET (RULE 26) dicyclohexylcarbodiimide (0.5? g, 2.75 mmol) gave 0.43 g (25$) 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

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) 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 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) 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 ~~ NHBoc 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) 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) WO 00/10565 PC'T/US99/03525 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

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) _99_ 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$) of the desired product as a white solid: 1H-NMR is consistent with structure; MS (FD) 492 (M+); Anal.
Calc' d for: G, 55.2:?; H, 6. 06; N, 14 . 86. Found: 55.33; H, 6.28; N, 13.24; IR (KBr) 3224, 3061, 3032, 2962, 2936, 2873, 1678, 1636, 1538, 1498, 1454, 1101 cm 1.
Preparation 49 H
O N NHBoc O O
HN
N
N
N
Reaction of a compound of Preparation 47 (1.00 g, 580 mmol) , hexamethylene~imine (0.2 mL, 1 .7 mmol) , 1-hydroxybenzotriazole hydrate (0.25 g, 1.9 mmol) and 1,3-dicyclohexylcarbodii.mide (0. 4 g, 1 . 9 mmol ) in N, N-dimethylformamide (50 mL) as described in Preparation 4 gave 0.76 g (68~) of the desired product as a tan foam: 1H-NMR is consistent with structure; MS (FD) 660.2 (M+); Anal. Calc'd for: C, 65.43; H, 7.32; N, 12.02. Found: C, 65.92; H, 7.86;
N, 11.71.
SUBSTITUTE SHEET (RULE 26) Example 12 H
~2 O O
HN
N
~,"~ N
2.67 HC1 Reaction of a compound of Preparation 49 (0.67 g, 1.0 mmol) and trifluoroacetic acid (4 mL, 52 mmol) in dichloromethane (12 mL) for 1 h at ambient temperature, followed by acidification with HC1 in ethyl acetate, according to Preparation 4 gave 0.3 g (48$) 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 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) WO 001105b5 PCT/US99/03525 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) acetate. The combined organic extracts were washed with water, brine, dried (Na2S04) and concentrated. Purification by silica gel chromatography (25$-75~ ethyl acetate/hexanes) gave 0.580 g (5090) of the product (mixture of diastereomers) as a yellow oil: 1H NMR (300 MHz, CDC13) d 8.72-8.73 (d, 1H, J = 2.lHz), 8.22-8.27 (dd, IH, J = 9.2Hz; 2.lHz), 8.15 (s, 1H), 7.44-7.50 (app. t, 3H, J = 6.9Hz), 7.34-7.41. (m, 3H), 6.19 (s, 1H), 4.26-4.39 (m, 2H), I.27-1.33 (t, 3H, J =
6.9Hz); FD+ MS for CI7H15N304 = 325; Anal. calcd. for C17HI5N304: C, 62.'T6; H, 4 . 65; N, 12, 92; Found: C, 62. 89;
H, 4.92; N, 12.92.
Preparation 52 HN
HO
i To a solution o f a compound of Preparation 50 (0.81 g, 2.73 mmol) stirring .in dioxane (30 mL) at room temperature was added LiOH~H20 (0.57 g, 13.6 mmol) and H20 (15 mL).
After 45 min, the mixture was concentrated to a volume of approximately 20 mL.. The resulting aqueous solution was diluted with H20 (7'i mL) and extracted with diethyl ether.
The aqueous layer was acidified with 1N HC1 and extracted with ethyl acetate. The combined organic extracts were washed with brine, dried (Na2S04) and concentrated to give 0.71 g (95$) 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) 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) Preparation 54 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$) of the product (l:l mixture of diastereomers) as a yellow solid: 1H NMR (300 MHz, CDC13) S 7.43-7.50 (m, 3H), 7.27-7.43 (m, 4H), 7.13-7.20 (app, t, 1H, J = 7.5Hz), 6.83-6.91 (t, 1H, J = 5.8Hz), 5.14 (s, 1H), 4.52-4.58 (m, 0.5H), 4.41-4.47 (m, O.SH), 3.89-3.97 (m, 1H), 3.71 (s, 1.5H), 3.62 (s, 1.5H), 3.23-3.36 (m, 1H), 1.82-2,24 (m, 5H); 13C NMR (75.5 MH2, CDC13) d 172.2, 171.7, 168.7, 168.5, 149.0, 146.9, 146.5, 136.4, 135.9, 129.5, 129.4, 129.0, 128.8, 128.5, 128.2, 128.0, SUBSTITUTE SHEET (RULE 26) 127.8, 119.9, 119.6, 112.2, 112.0, 106.5, 106.5, 59.5, 59.4, 59.3, 59.3, 52.2, 52.0, 46.7, 46.7, 28.7, 28.6, 24.9, 24.5;
FD+ MS for C2pH21N305 = 383; Anal. calcd. for C2pH21N305:
C, 62.65; H, 5.52; N, 10.96; Found: C, 61.93; H, 5.62; N, 10.46.
Preparation 55 OZN y_ N
~ /
O
O~O
I
to To a solution of a compound of Preparation 53 (0.43 g, 1.46 mmol), L-proline methyl ester hydrochloride (0.24 g, 1.46 mmol), 1-hydro:~cybenzotriazole hydrate (0.20 g, 1.46 mmol ) and N, N-diisopropylethylamine ( 0. 66 g, 5 . 10 mmol ) stirring in anhydrous 1,2-dichlormethane (30 mL) at room temperature was added 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (0.31 g, 1.60 mmol). After 18 h, the reaction mixture wa~~ quenched with H20 (50 mL) and extracted with ethyl acetate. The combined organic extracts were washed with 10~ citric acid, saturated aqueous sodium bicarbonate, H20, brine, dried (Na2S04) and concentrated.
Purification by radial chromatography (silica gel, 50~ ethyl acetate/hexanes to 100$ ethyl acetate gradient) gave 0.25 g (42$) of the a single diastereomer as a white foam solid:
1H NMR (300 MHz, CDC13) d 8.75-8.76 (b, 1H, J = 2.lHz), 8.28-8.32 (dd, 1H, J = 8.9Hz; 2.lHz), 7.91 (s, 1H), 7.45-7. 58 (m, 6H) , 6.26 (s, 1H) , 4 . 65-4 .70 (m, 1H) , 3. 83-3 . 92 (m, 1H), 3.78 (s, 3H), 3.30-3.39 (m, 1H), 1.95-2.30 (m, 5H); FD+
SU8ST1TUTE SHEET (RULE 26) MS for C21H20N405 =~ 408; Anal. calcd. for C21H20N405: C, 61.76; H, 4.94; N, 13.72; Found: C, 61.24; H, 5.16; N, 13.10.
Preparation 56 ~ H
''N ~ NHBoc C, ~~
O NI-r HN
N ~~ w Ow-y' O
O
To a slurry of 5$ 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$) of the product (mixture of diastereomers) as an off-white solid foam. 1H NMR (300 MHz, CDC13) d 8.83-8.91 (rn, 1H), 7.44-7.51 (m, 2H), 7.20-7,36 (m, SUBSTITUTE SHEET (RULE 26) WO 00/10565 PGT/tJS99/03525 8H), 6.88-7.06 (m, 3H), 6.32-6.38 (app. t, 1H, J = 6.9Hz), 5.28 (s, 1H), 5.12-5.19 (m, 1H), 9.88-4.91 (br. s, 1H), 4.48-4.60 (m, 3H), 4.17-4.24 (m, 1H), 3.64-3.72 (app. q, 2H, J = 8.OHz), 3.62 (s, 3H), 3.39-3.52 (m, 1H), 3.28-3.39 (m, 1H) , 1 . 81-2.15 (m, 5H) , 1 .53-1. 57 (app. d, 3H, J = 7. 9Hz) , 1.38 (s, 3H), 1.39 (s, 9H); FD+ MS for C39H4gN50g = 716;
Anal. calcd. for C3!aH4gN50g: C, 65.44; H, 6.90; N, 9.78;
Found: C, 65.23; H,, 7.43; N, 10.34.
Preparation 57 O ~ N~'NHBoc O
O N
i O ,..~ N
'' ''~1 N -'~
O O
To a slurry of 5$ 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) WO 00/10565 PC'f/US99/03525 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) Example 14 w ~~ O~"N~'NH 3+CF 3CO0 ' I ~ ~... o 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) WO 00/10565 PCTlUS99/035Z5 Preparation la ~~ NHBoc CO Me 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) 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) Preparation ld H
O~N NHBoc 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) 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 (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) WO 00/i056S PCT/US99/035Z5 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) WO 00/10565 PCTNS99l03525 (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

(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) 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) ( 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) 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~

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 WO 00/10565 PC'T/US99/03525 (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~

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) 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) WO 00/10565 PC'T/US99/03525 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) Examples 1 and 2 W. N N
O
i' ~~ O
O N
~N 2HC1 ~N N.J
/ ' ~ v 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$) of the desired product as a white solid: 'H NMR (d, DMSO) 0.50 (d, J = 6. 0 Hz, 1 . 5 Fi) , 0. 80 (d, J = 6. 0 Hz, 1. 5 H) , 1 .16 (m, 1H), 1.35 (m, 1Fi), 1.50-1.70 (m, 8H), 2.60-2.70 (m, 2H), 3.03 (m, 1H), 3.65-3.80 (m, 6H), 4.40 (m, 1H), 4.53 (s, 2H), 4.75 (m, 1H), 6.90-7.08 (m, 3H), 7.25-7.45 (m, 9H) , 8. 20-8 . 40 (m, 41:i) , 8 . 61 (d, J = 7. 5 Hz, 1H) , 11.15 (br s, 1H); tR = 7.93 min; MS (ion spray) 591.6 (M+1);
SUBSTITUTE SHEET (RULE 26) Anal . Calc' d for C;ZH42N6O5'2HC1: C, 57. 92; H, 6. 69; N, 12.66. Found: C, 57.72; H, 6.47; N, 12.42.
Example 2. Isomer 2 To a solution of the purified isomer in ethyl acetate was added a saturated solution of hydrochloric acid in diethyl ether. The resulting slurry was concentrated to yield 0.98 g (33$) 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$) of the desired product as SUBSTITUTE SHEET (RULE 2B) WO 00/t 0565 PCT/US99/03525 a tan foam: 'H-NMR (d, DMSO) 1.20-1.90 (m, 18H), 1.40-1.90 (m, 3H), 2.83 (m, 1H), 3.55-3.73 (m, 3H), 3.75 (s, 3H), 3.85 (m, 1H),. 4.45 (d, J - 3.8 Hz, 2H), 4.60 (m, 1H), 6.65 (d, J = :L0.93 Hz, 1H), 6.95-7.05 (m, 2H), 7.10-7.20 (m, 2H), 7.20-7.50 (m, 11H), 8.00-8.10 (m, 2H), 10.15 (br s, 1H); MS (FD) 798.7 (M+); Anal. Caic'd for C43H51FN60g: C, 64.65; H, 6.43; N, 10.53. Found: C, 64.38;
H, 6.48; N, 10.61.
Examples 3 and 4 \ N~N
\O
O N
F / ~N
\I N N
O O ~ \
O
Reaction of the product of Preparation 7 (0.53 g, 0.66 mmol) and trifluoroacetic acid (2 mL) in dichloromethane (6 mL) as described i.n Example 1 gave 0.34 g (74$) 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) 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) 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) 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

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$) of the desired product: I.40-1.60 (m, 6H), 'H-NMR (d, DMSO) 1.75-1.95 (m, 3H), 2.20 (m, 1H), 2.95 (m, 1H), 3.60-3.80 (m, 9H), 4.40 (m, 7_H), 4.50-4.55 2H),4.75 (m, IH), (m, 6.70 (s, 1H), 7.00 (t, J - 8.7 Hz, 2H),7.40-7.45 (m, SUBSTITUTE SHEET (RULE 26) 9H), 8.05 (m, 1H),, 8.20-8.30 (m, 3H), 8.55 (m, 1H), 10.95 (m, 1H); MS (ion spray) 621.5 (M+1); Anal. Calc'd for C32HqpNgO~~2.3HC1: C, 54.55; H, 6.05; N, 11 . 93. Found: C, 54.96; H, 5.81; N, 11.79.
Preparation 10 N
-NFIH o c O O
N
~O
O/
To a suspension of 5$ 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) 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

N
O
O
SUBSTITUTE SHEET (RULE 26) 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

O O
N ~2HC1 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~

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) concentrated. The resulting carboxylic acid was combined with pyrrolidine (0.710 g,10 mmol), 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 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 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) 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 '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) 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 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) 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 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$) of the desired product as an off white solid which was used without further purification.
Example 11 To a solution of the compound of preparation 19 (0.74 g, 1.15 mmol) stirring .at room temperature in dichloromethane (30 mL) was added triflouroacetic acid (10 mL). After 2 h, the mixture was concentrated and the residue treated with excess aqueous sodium bicarbonate.
The resulting mixturE~ was extracted with ethyl acetate and the combined organic extracts were concentrated. The SUBSTITUTE SHEET (RULE 26) WO 00/10565 PC'T/US99/03525 residue was purified by flash chromatography (silica gel, chloroform/methanol.) to provide 0.23 g (37$) 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 WO 00/105b5 PCT/US99/03525 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 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) Preparation 23 N"J
~O
O ~O\
O-O N N O

~~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) 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\

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$) of the desired product as a tan foam: 1H-NMR is consistent with structure; MS (ion spray) 721.5 (M+1); Anal. Calc'd for CsaHSZNsOe : C, 63 . 31; H, 7 . 2 7 ; N, 11 . 6 6 . Found : C, 63 .18 ;
H, 7.30; N, 11.60.
Example 12 \O N N
/ O
O N
~N 2HC1 J
.~N N
O ~Ov SUBSTITUTE SHEET (RULE 26) Reaction of the compound of Preparation 25 (0.5 g, 0.7 mmol and trifluoroacetic acid (2 mL) in dichloromethane (6 mL) as described in Example 1 gave 0.4 g (83$) 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

O N O
C %N
NJ
~ \ o\

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) Example 13 r'~O N N
/ O
O N

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$) of the desired mixture of isomers as a white solid: 'H-NMR is consistent with structure; MS (high res) calc'd for CZ9H39N6O6: 567.2931. Found: 567.2938 . Anal . Calc' d for CasHseNs~s'2hydrochloi-ic acid: C, 54.46; H, 6.30; N, 13.14.
Found: C, 54.67; H, 6.08; N, 13.00.
Preparation 27 Reaction of 4-trifluoromethylphenyl acetic acid (15.0 g, 73.4 mmol) and p-to.l.uenesulfonic acid (2.8 g, 14.7 mmol) in absolute ethanol (100 mL) as described in Preparation 1 gave 16.3 g (95$) 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) 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 Preparation 30 O N N O
O
O N O
'~ N
O N
O

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) 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$) of the desired product as tan foam: 'H-NNtR (d, DMSO) 1. 20-1. 35 (m, 15 H) , 3 . 60 (m, 1H) , 3. 65 (m, 1H) , 4 . 45 (d, J = 2 . 6 Hz, 2H) , 4 . 60 (m, 1H), 6.46 (s, 1H), 7.15 (m, 1H), 7.20-7.35 (m, 6H), 7.42 (m, 1H), 7.57-7.65 (m, 3H), 7.79 (d, J - 8.3 Hz, 2H), 10.25 (br s, 1H); MS (ion spray) 648.9 (M+1); Anal.
Calc' d for C31H36F'3r~5O-7: C, 57. 41; H, 5. 60; N, 10. 81.
Found: C, 57.31; H, 5.59; N, 10.53.
Preparation 32 N N O
/ O
O N O
~N
~NJ
O
CFs Reaction of the product of Preparation 31 (8.0 g, 12.3 mmol), 4-methylpiperi.dine (1.5 mL, 12.3 mmol), 1-hydroxybenzotriazole (1.83 g, 13.5 mmol) and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (2.8 g, 13.5 SUBSTITUTE SHEET (RULE 26) mmol) in N,N-dimethylformamide (150 mL) as described in Preparation 6 gave 7.33 g (81$) 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

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) 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

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) 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

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) 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) 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 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) 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

N N,.
O

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 $) of isomer 2. Example 17. (isomer 1) 'H-NMR (d, DMSO) 0- 77 (d, J = 6. 5 Hz, 1. 5H) , 0. 87 (d, J = 6. 0 Hz, 1 . 5H) , 15 1.00 (m, 1H), 1.32 (s, 3H), 1.50 (s, 3H), 1.50-1.70 (m, 2H) , 2. 72 (m, 1H) , 3. 00-3.30 (m, 4H) , 3 . 75 (m, 1H) , 4 . 05-4 . 33 (m, 3H) , 4 . 20 (m, 1H) , 4 . 78 (m, 1H) , 6. 94 (m, 3H) , 7.20 (s, 1H), 7.30-7.40 (m, 2H), 7.55-7.70 (m, 2H), 7.75-8.00 (m, 4H), 8.05-8.15 (m, 2H), 8.50 (m, 1H), 10.86 (s, 1H) , 11. 05 (s, 1H) ; 1=k = 6. O1 min; MS (ion spray) 638.2 (M+1 ) . Example 18. (~~.somer 2) rH-NMR (d, DMSO) 0. 77 (d, J
- 6. 5 Hz, 1. 5H) , 0. 8',~ (d, J = 6. 0 Hz, 1 . 5H) , 1 . 00 (m, 1H), 1.32 (s, 3H), 1.50 (s, 3H), 1.50-1.70 (m, 2H), 2.72 (m, 1H), 3.00-3.30 (m, 4H), 3.75 (m, 1H), 4.05-4.33 (m, 3H), 4.20 (m, 1H}, 4.78 (m, 1H), 6.94 (m, 3H), 7.20 (s, 1H), 7.30-7.40 (m, 2H:), 7.55-7.70 (m, 2H), 7.75-8.00 (m, ;5U8STITUTE SHEET (RULE 28) WO 00/10565 PC1'NS99/03525 4H), 8.05-8.15 (m, 2H), 8.50 (m, 1H), 10.86 (s, 1H), 11.05 (s, 1H); tR = 7.5 min; MS (ion spray) 638.2 (M+1).
Preparation 37 N O
~O
~N
r/
O N
.~ O N
O
C F~
A mixture of the product of Preparation 29 (11.1 8, 32.3 mmol) and 5$ 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$) of the desired product as an orange solid: 1H-NMR (d, DMSO) 1.15 (t, J = 7 Hz, 3H), 1..18-1.32 (m, 15H), 1.35-1.70 (m, 4H), 3.23 (m, 2H) , 4 .19 (q, J = 7 Hz, 2H) , 4 . 31 (m, 1H) , 6. 58 (s, 1H) , 7. 00 (br s, 1H) , 7. 05-7.22 (m, 6H) , 7.41 (m, 1H), 7.52-7.58 (m, 3H), 7.75 (d, J = 8 Hz, 2H), 10.19 (br SUBSTITUTE SHEET (RULE 26) s, 1H) ; MS ( ion spray) 674 . 7 (M+1 ) ; Anal . Calc' d for C39H42F3N506: C, 60. E~1.; H, 6.28; N, 10. 39. Found: C, 60. 44;
H, 6.48; N, 10.36.
Preparation 3B
N~O
~O
'' ~~N
O N
~N
N -"J
O
r CFa To a solution of tlhe product of Preparation 37 (12.0 g, 17. 8 mmol) stirring .in dioxane (20 ml) and water (20 ml) at room temperature was added lithium hydroxide (0.84 g, 35.6 mmol). After 90 min with intermittent sonication, the reaction was poured into a solution of sodium bisulfate (12 g/50 mI~ Hz0) and brine (20 ml,) then extrar_ted with ethyl acetate. The combined organic extracts were dried over sodium sulfate, filtered, and concentrated to provide 11.5 g (100$) 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) Preparation 39 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) 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) (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 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) 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

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

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) 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

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

N N O

~N
.SUBSTITUTE SHEET (RULE 26) 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

___CN N F
O

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) N N O-y( o N° o ~'N
NJ F

r 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' 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 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) Preparation 49 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) 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

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) 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) WO 00/10565 PC'T/US99/03525 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 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

~N N
O
F
SUBSTITUTE SHEET (RULE 26) WO 00/10565 PCT/US99i03525 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) 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) 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) 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) WO 00/10565 PC'T/US99/03525 Preparation 58 N N O
O
N I'O
C %N
N NJ

/ 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) wo oonos6s Pc~rmsmo3s2s 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) 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 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) 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

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) 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

-.'"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) Exempla 30 N N
_O
r o O N

N N
\ F

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~

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) WO 00/t0565 PCT/US99/03525 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

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) 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

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 Preparation 70 N, _O
r _o ~ ~~.~(o ~'. 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) 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) 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~

ale 34 O N N
_ O

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

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 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 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 WO 00/10565 PC"f/US99/03525 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) 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 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) 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 o = N:
N
~~~ 0 p ~ w 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 (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) 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

O N O
~N
J
N
N
O
O
SUBSTITUTE SHEET (RULE 26) O
--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

~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) 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) wo oonos6s Pcr~us99io3sZs 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
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) 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~

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

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) 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) (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~

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) 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

o'~
SUBSTITUTE SHEET (RULE 26) Preparation 88 Preparation 89 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. 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) 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 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 p.-N
--.

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 wo oon os6s Pcrius99iossis 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) 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) 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

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) N \ N' .O
~I.I(O
O
O N O
/ ~'.N
J
~N N
o / ~ i ~I

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 ..

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 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 / \
\ /
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) 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

SUBSTITUTE SHEET (RULE 26) 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) 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 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) 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) °/
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 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) N N
~O
O
O N
~N
/~ NJ
~N
F
O
Preparation 108 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 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

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) 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) 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) (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

/''~ J
--( ,N N F
O
SUBSTITUTE SHEET (RULE 28) 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 Example 51 O N N
/ O
O N

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) ~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 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 Preparation 119 N N O
~~O
/ /~ O
O
~C

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

~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) 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, SUBSTITUTE SHEET (RULE 26) 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 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 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) 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) 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
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) 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

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) 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) 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~

O

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) 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) 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 i p~N~
N
~-O

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) WO 00/10565 PC'T/US99/03525 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) 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

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) 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) WO 00/10565 PC'T/US99/03525 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

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) 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) WO 00/lOS6S PGT/US99/03S2S

(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) 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) WO 00/10565 PC'T/US99/03525 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) s t DEMANDES OU BREVETS VOLUMINEUX

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 IS VOLUME ~_ OF
MOTE: For additional voiumes~please canta'ct'the Canadian Patent Office

Claims (15)

We claim:

1. A compound of formula I

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, 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 and 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, 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 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

3. A compound according to Claim 1 wherein B is

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

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

7. A compound according to Claim 1 wherein E is

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 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.