Note: Descriptions are shown in the official language in which they were submitted.
CA 02195125 2000-10-10
-1_
5 DIFLUOROSTATONE ANTIVIRAL AGENTS
BACKGROUND OF THE INVENTION
10 Retroviruses are a class of viruses which transport
their genetic material as ribonucleic.acid rather than as
deoxyribonucleic acid. Retroviruses are associated with a
wide variety of diseases in man, one of which is AIDS.
Although there have been disclosures of other anti-viral
15 agents useful in the treatment of AIDS, for example see
patent applications EP 0 218 688, EP 0 352 000 and PCT/US
91/09741, the compounds of the present invention have not
been previously disclosed.
20
SUMMARY OF THE INVENTION
The present invention relates to compounds having try
following general formula (I);
25
R~ CNH-CH CNH-CH-C-CF2-C NR5R6
O O O O
x
30
formula I
and the stereoisomers, hydrates, isosteres and the
pharmaceutically acceptable salts thereof wherein
35
~l'~ ~1
wo ~Eroz.~9s rcz~usvstt~~aa~
_z_
P1 is
T
T'
C~~6 alkyfene
wherein T is [(O)b-W-R] and T' is [(O)b~-W'-R'~ or
hydrogen, wherein each of W and W' are independently
C1_~ alkylene or nothing,
14 rovided that W is C
P 2-s alkylene when W is directly
attached to a nitrogen atom in R,
provided that W' is C2_6 alkylene when W' is
directly attached to a nitrogen atom in R',
provided that W or W' are each independently C1_s
1' alkylene when R or R' are each independently an
aryl;
P2 is C1_b alkyl, cyclopentyl, hydroxy C1_~ alkyl,
phenyl, benzyl or 3-tetrahydrofuryl;
R and R' are each independently -CH2CH0, hydroxy Gx_g
alkyl, C1_6 alkoxy Ci_6 alkyl, C1_6 alkyl,
~R3)d ,
C1_~, alkenylene, piperazinyl, substituted piperazinyl,
piperidyl, morpholinyl, pyridyl, pyrazinyl, pyrimidinyl
or phenyl wherein substituted piperazinyl is
3D piperazinyl substituted on one nitrogen atom thereof
with CHO, C(O)ri7HR'q, C1_q alkyl or CO2Rq;
3~
~1'O ytt02~99 "o ~ " PC"TN695107241
Ll~v~l~'S
-3-
R1 is
\ \ \
~ N
/ (CHz)O- , NI~(CHz)O- , ~ / (CHz)O- ,
O O
I' N ~ \ hN I \
O_ J N~ ~ O\~ /
'~/O \% \ \/O
~S
Ni \ \ C02R~4
I / I / CH-O- ,
I \ IH O~ I \
/ (CHz)z-CH- ' ~N-CHZ- ' / CH~CH- ,
I \ OH O 1 ~~CHz-.
/ CH- ' ~/NI -CCHZ_ or
N ,
27
R3 is C1_6 allenyl C1_6 alkoxy, C1_6 alkylene,
hydroxy C1_6 alkyl, C1_6 alkyl or OH;
3G
Rq is Cz_6 alkyl, phenyl or 6enzyl;
R'q is hydrogen or C1_6 alkyl;
ti '; ~ ~ E_
VV09GI02.i99 ~ ~ ~ "~ : ") PCTlLS9il072d1
-~_
R5 is hydrogen, C1_15 alkyl, OH, hydroxy C1-15 alkyl,
-CH{((CH2)d-D-CHZ)x-R~e)2r -CH2Si{CH3)2(R3), PDL, _
(C1_6 alkylene)-ORq, -CH(Y)(Z),
(v)e HO
C~_salkylene~ ' (CH2)b
(a) (b)
N ~.r
- ~~ or CHz~ \ a
\N
(c) (d)
wherein PDL is -(CHz)e-2-, 3- or 4-pyridyl, or p-
substituted benzylaxy, wherein the substitution is with
a vitro, OH, amino, C1_s alkoxy, hydroxy C~_6 alkylene,
or halogen: Y is C1_15 alkyl, hydroxy Cl-lg alkyl, C1-6
alkyl or -(CH~)e-C6Hq-(V)~': Z is -(CHZ)d-O-CHO, CI-s
alkyiene-O-(CHy)d-(O-CHZ-CHZ)e-O-C1_6 alkyl, CHO, COZRq,
CO2NHRq, -(CHZ)a-O-(CH2)awR'~. -(CHa)e-ORq or
~(V)e
(CH2)e
(e)
wherein V is ORq or hydroxy C1_6 alkylene:
provided that d'=2 when R'7 is giperazinyl, substituted
piperazinyl, giperidyl or morpholinyl:
35
2i~i~~%5
i4'0 9610:499 PCTlUS95107241
_5_
R6 is as defined for RS with the proviso that R6 is
other than hydrogen when RS is hydrogen, or RS and R6
are taken together with the nitrogen atom to which they
are attached are selected from the group consisting of;
-N , - , -N~ ,
to (~ (9? (h)
(tHz?b / R3
R3-Si
~N
3
15 (i)
8
20 -N~ -MHO , _N or -N~ R$
(i} (m) (n1
R7 is CH20R4, C(O)NHR4 or CHO;
R'7 is piperazinyl, substituted piperazinyl, piperidyl,
morpholinyl, pyridyl, pyrazinyl, pyrimidinyl or phenyl,
wherein substituted piperazinyl is piperazinyl
substituted on one nitrogen atom thereof with CHO,
C(O)NHR4, C1_4 alkyl or C02R4;
Rg is (H, OH) or =O;
R'e is pyrimidyl, pyridyl, pyrazinyl or phenyl;
-~ ~ \~ ~~ " ~ r
W096lL)2J9<3 ._ f ~;~1 ~ ~'~ PCTIiJS9~!Q7231
-6-
a is zero, 1, 2 or 3;
b and b' are each independently zero or L;
d and d' are each independently 1 or 2;
a and e' are each independently zero, 1 or 2; and ,
x is zero or one.
LED DESCRIPTION OF THE IN57ENTION
l~ The terms "halo", "halogen" or "halide" refer to a
chlorine, bromine or iodine atom.
Isosteres of the compounds of Formula I include those
wherein (a) the a-amino acid residues of the P1 and Pa
l~ substituents are in their unnatural configuration (when
there is a natural configuration) or (b) when the normal
peptide amide linkage is modified, such as for example, to
form
a
ao n
-CH2NH- (reduced), -C-N(CH3) (N-methylamide), -COCH2-
(keto), -CH(OH)CH2- (hydroxy), -CH(NH2)CHZ- (amino),
-CH2CHZ- (hydrocarbon). Preferably a compound of the
invention should not be in an isosteric form. Unless
otherwise stated the a-amino acids are preferably in their
L-configuration.
A compound of the invention may be in free farm, e.g.,
amphoteric farm, or in salt, e.g., acid addition or anionic
salt, form. A compound in free form may be converted into a
3 f~
salt form in an art-known manner and vice-versa.
The pharmaceutically acceptable salts of the peptide of
Formula I (in the form of water, or oil-soluble or
dispersible products) include the conventional non-toxic
3~
salts or the quaternary ammonium salts of these peptides,
which are farmed, e.g., from inorganic or organic acids or
bases. Examples of such acid addition salts include
WO 96102~t99 ~ ~ PCTIilS95J07241
acetate, adipate, alginate, aspartate, benzoate,
benzenesulfonate, bisulfate, butyrate, citrate, camphorate,
camphorsulfonate, cyclopentanepropionate, digluconate,
dodecylsulfate, ethanesulfonate, fumarate, glucoheptanoate,
glycerophasphate, hemisulfate, heptanoate, hexanoate,
. hydrochloride, hydrobromide, hydroiodide, 2-hydroxyethane-
sulfonate, lactate, maleate, methanesulfonate, 2-naphthal-
enesulfonate, nicotinate, oxalate, paemoate, pectinate,
persulfate, 3-phenylpropionate, picrate, pivalate,
propionate, succinate, tartrate, thiocyanate, tosylate, and
undecanoate. Base salts include ammonium salts, alkalimetal
salts such as sodium and potassium salts, alkaline earth
metal salts such as calcium and magnesium salts, salts with
organic bases such as dicyclohexylamine salts, N-methyl-
D-glucamine, and salts with amino acids such as arginine,
lysine, and so forth. Also, the basic nitrogen-containing
groups may be quaternized with such agents as lower alkyl
halides, such as methyl, ethyl, propyl, and butyl chloride,
bromides and iodides; dialkyl sulfates like dimethyl,
diethyl, dibutyl; and diamyl sulfates, long chain halides
such as decyl, lauryl, myristyl and stearyl chlorides,
bromides and iodides, aralkyl halides like benzyl and
phenethyl bromides and others.
The hydrates of the compounds of Formula I are hydrated
compounds having the partial structure
~ pi~CF2~~~
HOxOH O
and in their end-use application are generally the active
forms.
21 X51 ~_'S
W 0 96102.199 PCT)IiS95)O i 241
_8-
In general, as used herein, the term "alkyl" includes
the straight, branched-chain and cyclized manifestations
thereof unless otherwise indicated, particularly such
moieties as methyl, ethyl, isopropyl, n-butyl, t-butyl,
-CH -t-but 1, c clo ro 1, n ~'
y y p py -propyl, pentyl, cyclopenty~,
n-hexyl, cyclohexyl and cyclohexylmethyl. The term
"aralkyl", when used, includes those aryl moieties attached
to an alkylene bridging moiety, preferably methyl or ethyl.
«Aryl" includes both carbocyclic and hetereocyclic
moieties of which phenyl, pyridyl, pyrimidinyl, pyrazinyl,
indolyl, indazolyl, furyi and thienyl are of primary
interest; these moieties being inclusive of their position
isomers such as, for examgle, 2-, 3-, or 4-pyridyl, 2- or
3_furyl and thienyl, 1-, 2-, or 3-indolyl or the 1- and 3-
indazolyl, as well as the dihydro and tetrahydro analogs of
the furyl and thienyl moieties. Also included within the
term "aryl" ors such fused carbocyclic moieties as
pentalenyl, indenyl, naphthalenyl, azulenyl, heptalenyl,
acena hth ten 1 fluaren 1 henalen 1 henanthren 1,
P Y Y . Y . p Y ~ p Y
anthracenyl, acephenanthrylenyl, aceanthrylenyl,
triphenylenyl, pyrenyl, chrysenyl and naphthacenyl. Also
included within the term "aryl" are such other heterocyclic
radicals as 2- or 3-benzo[b]thienyl, 2- or 3-naphtho[2,3-
b]thienyl, 2- or 3-thianthrenyl, 2H-pyran-3-(or 4- or
5-)yl, 1-isobenzo- furanyl, 2H-chromenyl-3-y1, 2- or 3-
phenoxathiinyl, 2- or 3-pyrrolyl, 4- or 3-pyrazolyl,
2-pyrazinyl, 2-pyrimidinyl, 3-pyridazinyl, 2-indolizinyl,
1-isoindolyl, 4H-quinolizin-2-yl, 3-isoquinolyl, 2-
~inolyl, 1-phthalazinyl, 1,8-naphthyridinyl, 2-
quinoxalinyl, 2-quinazolinyl, 3-cinaolinyl, 2-pteridinyl,
4aH-carbazol-2-yl, 2-carbazolyl, 8-carbolin-3-yl,
3-phenanthri.dinyl, 2-acridinyl, 2-perimidinyl,
1-phenazinyl, 3-isothiazolyl, 2-phenothiazinyl,
3~ 3-isoxazalyl, 2-ghenoxazinyl, 3-isochromanyl, 7-chromanyl,
2-pyrrolin-3-yl., 2-imldazolidinyl, 2-imidazolin-4-yl,
2-pyrazolidinyl, 3-pyrazolin-3-yl, 2-piperidyl,
21~~1~~
W096/(12999 PCTIt;S9S1072d1
_g_
2-piperazinyl, 1-indolinyl, 1-isoindolinyl, 3-morpholinyl,
benzo[b]isoquinolinyl arid benzo[b]furanyl, including the
position isomers thereof.
Likewise the term "alkylene" includes straight or
branched-chain moieties. Some examples of branched-chain
alkylene moieties are ethylethylene, 2-methyltrimethylene,
2,2-dimethyltrimethylene, and so on. For example, C3
alkylene can mean
CH3
-CH2-CH2-CH2- or -C- or -CH2-CH- or -CH-CH2-
I I I
CH3 CHI CH3
1~ All (C1_s) moieties such as C1_s alkyl, C1_s allenyl, C1_s
alkoxy, and hydroxy C1_s alkyl, are more preferably C1_3
moieties (containing 1-3 carbon atoms instead of 1-6 carbon
atoms).
The fluorenylmethyloxy moiety is that moiety generally
called by its abbreviation FMOC, and is the fluorenyl
moiety bearing -CH20 attached to the 9-position of the fluo-
renyl moiety. Other terms defined herein are piperazinyl
~-N,~ H~ or substituted piperazinyl ~ N~ -*~
the substitution (*) occurring only at one nitrogen atom
which is not attached to the remainder of the molecule
(attachment via a nitrogen atomj. The substituents are one
of CHO, C(O)NHR4, CI_y alkyl or C02R4.
The term "Bn" refers to a benzyl functionality of the
formula;
CHz
wo g~!o:a~~ ~ j ~ '~ a L 5 pc:~rrusosroaza l
-lo-
Piperidinyl and morpholinyl both bind to the rest of
-N~ -NCO
molecule via their respective nitrogen atoms while
pyrimidinyl, pyridyl and pyrazinyl bind to the rest
N~ N N
of t/he molecule anywhere except their respective
nitrogen atoms.
1~ Mare specifically, in the instance wherein P2 is either
Cl_6 alkyl or hydroxy Cy_6 alkyl, such moieties as -C(CH~)3,
-CH{CH3)Z, -CH(CH3)(CZHS), -C(OH)(CH3)3 and -CH(OH)CH3 are
preferred. The "hydroxy Cl_6 alkyl" moiety is illustrated in
one example by -CHZ-OH, the "C1_6 alkoxy Cl_6 alkyl" moiety,
is illustrated in one example by -CH2-OCH3, (although in
each instance the Cl_6 alkylene may be straight or branched
and the hydroxy radical is not limited to the terminal
carbon atom of the alkyl moiety).
2a As it is often quite advantageous to have what is
termed an amino protecting group (Pg), the scope of those
compounds of Formula I includes those Rl moieties which,
together with their adjacent carbonyl moiety farm such
groups as acetyl (AC), succinyl (Suc), benzoyl (Bz),
3p t-butyloxycarbonyl (Boc), benzyloxycarbonyl (CBZ), tosyl
(TS), dansyl (DNS), isovaleryl (Iva), methoxysuccinyl
(MeOSuc), i-adamantanesulphonyl {AdS02), i-adamantaneacetyl
(AdAC), phenyiacetyl, t-butylacetyl (fiba), bis[(1-
naphthyl)methyl]acetyl (BNMA) and Rz wherein Rz is an aryl
3y group as previously described suitably substituted by 1 to
3 members selected independently from the graup consisting
of fluoso, chloro, bromo, iodo, trifluoromethyl, hydroxy,
alkyl containing from 1 to 6 carbons, alkoxy containing
~ wo y~ni:~v~ 2 ~ ~ ~ ~ ~ 5 rcTm~svS~a~aai
-11-
from 1 to 6 carbons, carboxy, alkylcarbonylamino wherein
the alkyl group contains 1 to 6 carbons, 5-tetrazolo, and
acylsulfonamido (i.e., acylaminosulfonyl and sulfonylamino-
carbonyl) containing from 1 to 15 carbons, provided that
when the acylsulfonamido contains an aryl, the aryl may be
further substituted by a member selected from fluoro,
chloro, bromo, iodo and nitra.
Among the classes of amino protecting groups
contemplated are: t1) acyl type protecting groups such as
formyl, trifluoroacetyl, phthalyl, p-toluenesulfonyl
(tosyl), benzenesulfonyl, nitrophenylsulfenyl,
tritylsulfenyl, O-nitrophenoxyacetyi, and a-chlorobutyryl;
t2~ aromatic urethane type protecting groups such as
benzyloxycarbonyl and substituted benzyloxycarbonyls such
as p-chlorobenzyloxycarbonyl, p-methoxybenzyloxycarbonyl,
p-nitrobenzyloxycarbonyl, p-bromobenzyloxycarbonyl,
1-(p-biphenylyl)-1-methylethoxycarbonyl, a-, a-dimethyl-
3,5-dimethoxybenzyloxycarbonyl, and benzhydryloxycarbonyl;
(3) aliphatic urethane protecting groups such as tert-
butyloxycarbonyl (Boc), diisopropylmethoxycarbonyl, iso-
propyloxycarbonyl, ethoxycarbonyl, and allylaxycarbonyl;
C4) cycloalkyl urethane type protecting groups such as
cyclopentyloxycarbonyl, adamantyloxycarbonyl, and cyclo-
hexyloxycarbonyl; <S) thio urethane type protecting groups
such as phenylthiocarbonyl; t6> alkyl type protecting
groups such as triphenylmethyl (trityl) and benzyl (Bn);
t7) trialkylsilane protecting groups such as trimethyl-
silane if compatible. The preferred a-amino protecting
groups are tert-butyloxycarbonyl (Boc) or benzyloxycarbonyl
(CBZj. The use of Boc as an a-amino protecting group for
amino acids is described by Bodansky et al. in "The
Practice of Peptide Synthesis", Springer-verlag, Berlin
(1984), p. 20.
The term "stereoisomers" is a general term for all
isomers of individuals molecules that differ only in the
orientation of their atoms in space. It includes mirror
r} j ~ G s J! ~
CVO 9Gt02499 4 s ~ ~ '. ') PCTlU595l07241
-12-
image isomers (enantiomers), geometric (t1s/trans) isomers,
and isomers of compounds with more than one chiral center
that are not mirror images of one another
(diastereoisomers). Far amino-acids, the designations L!D,
or R/S can be used as described in IUPAC-IUB Joint
Commission on Biochemichal Nomenclature. E~an~l.Biachem. 138: ,
9-37 (i984).
In general the compounds of this invention may be
prepared using standard chemical reactions analogously
known in the art. More specifically, the preparation of
compounds of structure (3) is well known in the art and
described generally by Schirlin, D. and Van Dorsselaer, V.
in PCT/U591/09741 published July 23, 1992 with an
international publication number of WO 92/12123.
The compounds of formula (I) can be grepared as
described in Reaction Schemes A, A', A", B, C and D. All
the substituents, unless btherwise indicated, are
previously defined. The reagents and starting materials
are readily available to one of ordinary skill in the art.
30
WO 9b102~99 ~ ~ ~ ~ ~ ~ ~ PCTIfJS95107241
13
a
O
U=
O Z
n n Z
N ~ U= O n 'O I
v ~ ~' v U- O n
U n N ~
M W ~ N t!1
U- n-I ~ U v W r1
O
n " x x V v
i
~y- U- O x x
x
U- O
x
T W-U o T
a- a
a x
Z N
x
z
..
.r
v
z
c
z
z
z
U- O Z
'' N i
W U= O
O W
U= U ~ N
O i
W .C x x 'O W
U n v U- O n v v n
m ~ ~ ,_, ~ ,n x x ov
x
.i " _ ~ U- O ,,,
U- - co
O
a .a
- S
c, a
v a
z -
~
N
a
x
w v
z
v
n
U
v
n
Z
v
C b
N
Z a'
U= p x
O ~N u= O
S W I
V U cv N
_ _ G.
V1 _
V
a-U P1 U- O vp 1
1
x
'r ~ ~ V- O
b
a
N m ~ a-~
z
C
a s
W z
mt a u, o m,
ri r1 N N
~~~~j~~.~7
wo ~sor?.syy ec~rausv~omai
14
a
z
a
cr= o
1
N
W
U
1
U
- p
p,- V
n 1
n
Z a0
.H ~ V= O
t v
N S
w-v
t
a
t
v=o
.a
m
z
z
x ~o
U
= D
N
N W
W V
V '
V= O
a '
~- a b
v
S
5
w V a
C
1
Z n C>= t,
O
t
' O N
~ ~
- O
d ; .-K G-
~ v N = v n
T ~ w - V Z
v 1
1
S N Uy O
a
n
V=O V 'y.
N
C
v
..
z x
x
~
L V~ O
1=O
1 !
N N
W W
V V
m x ci=
o
V- O i
C7 .y ~ .i x
m w- V
-. v w- ,~ n
a C
n ~ 1 ~ x
p v x v. ~ ~ ~"~
U y "t w= O
iJl U= O
v x
N
'T.a N S w- V
O w-U
m
H
z
U 1 V= O
V - O - ~-t
N
S rn
w' N
W0961(12t99 ~ ~ ~'j ~ ~ ~ ~ PCTlLiS95/p7241
a
a
z
U= O
N
n W
G1 U n
u- O '~
v .~. yr
o s
w- a
i
m
x
u- o
x
a
N
a
z a
U= O a
Z
N v= o
w
U N
1 w
O
U- Y
n .i ~ V= O
V
N r,
n v p,-
v S
Z
U= p ,~ n
.w N Uc O
a V s~
v a
n
U
00
v
w
a
a
a
a
z V=O
V-p
N
By N
W
V U
G7 ra r U- O V= O
~
~ ~ a
a
a
v ~ n
n s
v Z
H , n
E., v= o ~ o
a
C ~ a
w
m
's' ~ o w
N N
~i t
WC79Gr02~99 L f l ~ ~ ~) Pl:'TlUB9510724C
-16-
In Scheme A, step (a) the aldehyde of formula (3) is
subjected to a condensation reaction under Reformatski
conditions with an. ester of bromodifluoroacetic acid,
preferably the ethyl ester in the presence of zinc and in
an anhydrous aprotic solvent, e.g., tetrahydrofuran, ether,
dimethoxyethane and the like under a nitrogen or argon
inert atmosphere. The reaction is gently heated to about
60°C for about 1-12 hours or ultrasonicated to produce
compounds (4).
Alternatively, in Reaction Scheme A step (a), the
condensation to produce compounds (4) can be achieved in
greater yields and at lower reaction temperatures utilizing
the following general method. Under an inert atmosghere,
such as nitrogen, the aldehyde (3) is dissclved in a
suitable anhydrous organic solvent. Examples of a suitable
anhydraus organic solvent are tetrahydrofuran, diethyl
ether, t-butyl methyl ether and the like. The solution is
cooled to apgroximately 0°C. To the solution is added about
0.30 equivalents of silver acetate, about 2.I equivalents
of zinc dust, and about 2 equivalents of ethyl
bromodifluoroacetate. About 0.34 equivalents of
diethylaluminum chloride (as a solution in toluene) is
added slowly to the reaction keeping the temperature of the
reaction below 12°C. The reaction is allowed to stir for 1
to 3 hours at about 0°C and then at room temperature for 4
to I2 hours. The reaction is then cooled to about 10°C and
quenched with saturated aqueous ammonium chloride. The
compound (4) is then isalated and purified by techniques
well known in the art. For example a solution of sodium
hydrogen tartrate is added and the reaction is allowed to
warm from 10°C to room temperature. The mixture is
filtered, the solids washed with a suitable organic
solvent, such as ethyl acetate and the layers of the
filtrate are separated. The aqueous layer is extracted
with ethyl acetate, the organic layer and extracts are
WO 9G/02499 ~ ~ ~ 'J ~ ~ ~ PCTlUS95107241
-17-
combined, dried over anhydrous magnesium sulfate, filtered
and concentrated. The residue is purified by flash
chromatography on silica gel with a suitable eluent, such
as cyclohexane/ethyl acetate to provide the compounds (4).
In Scheme A, step (b) formation of compounds (5) or
(I4) may be effected directly or undirectly. In one method,
the esters of formula (4) or (13) are de-esterified using a
suitable base, such as LiOH, KOH, NaOH and the like, in the
presence of water and a partially water miscible solvent
(such as tetrahydrofuran, dimethoxyethane, dioxane) at
about room temperature. The resulting acid can then be
aminated with the appropriate RSR6-substituted amine using
standard peptide-like coupling conditions. The selection of
the appropriate coupling reaction procedure is within the
skill of the art. The coupling reaction can be carried out
using standard coupling procedures such as the azide
method, mixed carbonic acid anhydride (isobutyl
chloroformate) method, carbodiimide
[dicyclahexylcarbodiimide, diisopropylcarbodiimide, or
water-soluble carbodiimide,l-(3-dimethylaminopropyl)-3-
ethylcarbodiimide hydrochloride (EDC)] method, active ester
(p-nitrophenyl ester, N-hydroxy-succinic imido ester)
method, Woodward reagent K method, carbonyldiimidazole
method, phosphorus reagents such as BOP-C1, or oxidation-
reductian methods. Some of these methods (especially the
carbodiimide method) can be enhanced by adding
hydroxybenzotriazole (HOST), for example the mixed
anhydride method may be employed, using DCC and
hydroxybenzotriazole at room temperature in solvents such
as CH2C12, tetrahydrofuran or dimethylformamide.
Alternatively the esters (4) or (13) may be directly
subjected to a reaction with the appropriate RSR6-
substituted amine without or with a solvent (tetrahydro-
furan) at a temperature of from 0 to 80°C.
vfO 9GJ02~i~ ~ ~ ~ ~ ~ ; ~ PCT/US9it072i1
-1$-
Alternatively, an appropriate RS, R6-substituted amine
that is protected as necessary is dissolved in a suitable
organic solvent, such as dichloromethane under an inert
atmosphere, such as nitrogen. An equivalent of a 2M
solution of trimethylaluminum in toluene is added dropwise
to the solution. After approximatelg 15 minutes this
solution is added to approximately 0.3 equivalents of ester
(4) or (13) dissolved in a suitable organic solvent, such
as dichloromethane. The reaction is allowed to stir for
about 15 to 24 hours at about room temperature to 40°C. The
product is then isolated using techniques well knowa in the
art. For example cold dilute aqueous hydrochloric acid and
ethyl acetate is added. The organic layer is separated and
washed with water, brine, dried over anhydrous magnesium
sulfate, f-filtered and concentrated under vacuum to provide
the compounds (5) or (14).
In Step (c) compounds (6), (8) or (11) are prepared by
removal of the P'1 protecting group using standard
procedures well known in the art [see T.H. Green,
"Protective Groups in Organic Synthesis", John Wiley and
Scns, 1981), such as hydrogenation. The free phenol
functionality is then reacted with an appropriate alkyl
halide in an inert solvent (preferably anhydrous dioxane,
anhydrous acetone or anhydrous dimethylformamide) in the
presence o~ a base (potassium or cesium carbonate) with or
without potassium iodide at room or reflux temperature.
In Step (c1) compound (13) is prepared by removal of
the P'1 protecting group using standard procedures well
known in the art[see T.H. Green, ~Protective Groups in
Organic Synthesis", John Wiley and Sons, 1981;, such as,
hydrogenation, P~~ being the compound obtained. Poe being a
free phenol.
In Step (c2) compounds (6), (8) or (11) are prepared
from the PoHderivatives (14), (16) or (17) by reaction with
WO 96!02-f99 C' a ~ ', PCTIUS95107i41
21 d..),~~
_19_
an appropriate alkylhalide in an inert solvent, in the
presence of a base. For example, the PoHderivative is
dissolved in a suitable organic solvent, such as acetone.
Approximately 1.2 equivalents of a suitable base, such as
potassium carbonate, are added followed by addition of
approximately 1.15 equivalents of the alkyl halide. A
catalytic amount of potassium iodide is then added and the
reaction is stirred for 1 to 3 days. The product is
isolated and purified by techniques well known in the art,
such as extractive methods and recrystallization. For
example, the reaction is poured into a suitable solvent
mixture, such as ethyl acetate/dilute aqueous sodium
chloride and the organic layer is separated. The organic
layer is then washed with dilute aqueous potassium
hydroxide, brine, dried over anhydrous magnesium sulfate,
filtered and concentrated under vacuum. The residue is
purified by recrystallization from a suitable solvent
mixture, such as cyclohexane/ethyl acetate to provide
compounds (6), (8) or (11).
In Step (d), for the preparation of Compounds (7}, (9)
and (15), the protecting groups Pg may readily be removed
by standard procedures well known in the art [see T.H.
Green, "Protective Groups in Organic Synthesis", ,john Wiley
and Sons, 1981], preferably acid/base hydrolysis (e. g.,
formic acid at room temperature followed by extraction of
the free base after treatment with sodium carbonate).
In Steg (e), Compounds (7), (9) or (15) are subjected
to a peptide coupling procedure with an appropriately
protected acid of the formula R'1CONHCH(Pa)C02H or R'lCOzH,
using the herein-described procedures (or by any other
coupling procedure well known in the art, or as described
in European Patent Application, Serial Number 93 401 785.6)
to produce compounds (8) and (11) (from compound (7)); (10)
and (12) (from compound (9)); and (16) and (I7) (from
compound (15)). R'1 is defined as R1 except far those
R'O 96ti)249'~ L ~ ~ r) ~ ~ .~ PCTliJS95/073d1
-20-
instances wherein protection of the R1 group may be required
as recognized by one of ordinary skill in the art. Far
example, a hydroxyl functionality on the alkyl portion of
the R1 group must be protected prior to the oxidation in
step (f).
In Step (f), the oxidation of compounds (8) and (11)
may be effected by methods well known in the art, such as
the Sworn oxidation procedure, or with 1,1,1-triacetoxy-1,1-
dihydro-1,2-benziodoxol-3(1H)-one to provide compounds of
formulas IA and IB.
In general the Sworn oxidation (see Synthesis, (i981),
165] is effected by reacting about 2 to 20 equivalents of
dimethylsulfoxide (DMSO) with about 1 to 10 equivalents of
trifluoroacetic anhydride ((CF3C0)20] or oxalyl chiaride
((COC1)z], said reactants being dissolved in an inert
solvent, e.g., methylene chloride (CHZC12), said reaction
being under an inert atmosphere (e.g., nitrogen or
equivalently functioning gas) under anhydrous conditions at
temperatures of about -70°C to -30°C to form an insitu
sulfonium adduct to which is added about 1 equivalent of
the appropriate alcohals, i.e., compounds (8) and (11).
Preferably, the alcohols are dissolved in an inert solvent,
e.g., CHyCl2, tetrahydrofuran, or minimum amounts of DMSO,
and the reaction mixture is allowed to warm to about -50°C
or -20°C (for about 20-60 minutes) and then the reaction is
completed by adding about 3 to 30 equivalents of a tertiary
amine, e.g., triethylamine, diisoprapylethylamine, /Y-methyl
morgholine, etc.
Alternatively the oxidation can be carried out with the
Doss-Martin periodinane (i.e., 1,1,1-triacetoxy-1,1-
dihydro-1,2-benziodoxol-3(1H)-one), [see Dess Martin, J.
Ors. Chem., 48, 4155, (1983)]. This oxidation is effected
by contacting about 1 equivalent of the alcohol with 1 to
10 equivalents of periodinane (preferably greater than 5
wo y6~ozan~> ~ ~ ~ ~ ~ ~ 5 PcTnrs9s~o~zai
-21-
equivalents), said reagent being in suspension in an inert
solvent (e. g., methylene chloride) under an inert
atmosphere (preferably nitrogen) under anhydrous conditions
at 0°C to 50°C (preferably room temperature) and allowing
the reactants to interact for about 1 to 48 hours.
Optional deprotection of the amine protecting groups may be
effected as desired after the ketones have been isolated.
In general, the modified Jones oxidation procedure may
conveniently be effected by reacting the alcohols with
pyridinium dichromate by contacting the reactants together
in a water-trapping molecular sieve powder, e.g., a
grounded 3 Angstrom molecular sieve), wherein said contact
is in the presence of glacial acetic acid at abaut 0°C to
50°C, preferably at room temperature followed by isolation
and then optionally removing amine protecting groups.
Alternatively, 1 to 5 equivalents of a chromic
anhydride-pyridine complex (i.e., a Sarett reagent prepared
insttu} [see Fieser and Fieser "Reagents for Organic
Synthesis" Vol. 1, pp. 145 and Sarett, et al., J.A.C.S. _25,
422, (1953)] in an inert solvent (e.g., CHyClz) under an
inert atmosphere under anhydrous conditions at 0°C to 50°C
is treated with 1 equivalent of the alcohol. The reaction
is allowed to stir for about 1 to 15 hours. The product is
isolated and purified by techniques well known in the art
followed by optionally removing amine protecting groups.
In step (g) the compounds decribed by formulas IA and
IB wherein R'1 is protected as required, are deprotected
under conditions well known in the art [see T.H. Green,
"Protective Groups in Organic Synthesis", John Wiley and
Sons, 1981], to provide compounds of formula IA' and IB'
wherein R"1 are those substituents which required
protection as required. For example the protected compound
is dissolved in a suitable organic solvent, such as
methylene chloride and treated with TFA/H20. The reaction
?i~%1~~5
V4'0 96f0:.199 PCTILT89514'72d t
_22_
is allowed to stir at room temperature for about 4 to 10
hours and then it is concentrated under vacuum. The
residue is purified by techniques well known in the art,
such as extractive methods followed by flash chromatography
(silica gel, hexane/ethyl acetate) to provide the
deprotected compound.
For the preparation of the necessary aldehydes of
formula (3) and the acids which are to be coupled with the
1G compounds (7), (9) or (15), alternative alkyTation
procedures are utilized depending upon whether the F1
and/or the PZ moieties are or are not residues of natural
amino acids. For the preparation of these intermediates
wherein the P1 or P2 moieties are residues of natural amino
15 acids (or manor modifications thereof, e.g.. P1 or Pz being
a benxyl or methyl ether of tyrosine)r the compounds are
either known or are prepared by processes and techniques
well known in the art.
20 To prepare the intermediates of the formula
P3
PgEiN-CHCOZRy
wherein Pg is an amino protecting groupr F3 is either a P'1
25 or P'2 moiety with P'1 and P'2 being as defined for PZ and
Pz respectively, except that they are other than residues
of naturally occuring amino acids, and the Rg moiety is an
alkyl radical, preferably methyl when P3 is P'~, and ethyl
when P~ is P'2, alternative methods are available.
To prepare the intermediates of formula
wo ~mro2a9!> 2 ~ ~ ~a ~ ~ ~ Pc~rn~s~sro~2ai
-23-
P.1 P.2
PgHN-CHCHO PgHN-CHCOOH
(1DB) (10A)
the following reaction scheme may be utilized
&EACTION SCHEME B
P3
PgNHCH2C02R9 (1) Base P9NHCHC02R9
(18) (2) P3X (19)
wherein P3 is as previously defined and X is a leaving
group, preferably halo or triflate, R9 is methyl when F3 is
P'1, and ethyl when P3 is P'2.
rn essence, the preparation of compounds (19) utilizes
the Krapcho method [Tetrahedron Letters, 26, 2205 (1976)]
for alkylation wherein compounds (18) are treated with a
base, e.g., LDA, (lithium diisopropylamide), followed by
reaction with the desired P3X in the presence of TMEDA
(i.e. tetramethylethylenediamine) in a solvent (tetrahydro-
furan) with or without HMPA (i.e. hepamethylphosphonamide)
according to the standard Krapcho conditions. Following
alkylation the compounds are then subjected to a reduction
using diisobutyl alaminum hydride (Dibal) in a mixture of
solvents, e.g., ether, toluene, hexane, tetrahydrofuran at
about -78°C for about 1 hour. Following the preparation of
the aldehydes of Formula (10B), the compounds are subjected
to the processes of Reaction Schemes A, A' and/or A".
Alternatively, the compounds of (19) may be prepared by
a Malonate/Curtius type sequence of reactions, [see Yamada,
et al., .T. Amer. Chem. Soc., (1972) 94, 6203] as illustrated
by the following reaction schema
WO 9fiIfB2-i99 ~ ~ ~ "~ l ~ ~ PCTfUS9id07241
-24-
fiEACTTON SCHEME C p
la
t-BUOaCCH2COaR9 t-Buo2CCHICOZR9
( 1 } Base ~
(20)
(2} P X (21)
ttemoval of t-Bu
P3
I
HOZCCHZCOZR9
(22)
Curtius-type
rearrangement
(19)
wherein t-BU is t-butyl, although other selectively
removable acid protecting groups may be utilized, and P3X
is as previously defined. This reaction involves the
al:cylation of the malonate ester (20) followed by selective
removal of the t-butyl protecting group to produce
compounds (22). These compounds are then transformed to
(19) using the Curtius type rearrangement which entails
their conversion to the protected amine via the
intermediately farmed acyiazides and isocyanates. The
resultant amines are then protected with standard amino
protecting groups, preferentially being protected insitu.
In the instance wherein Pa represents a P'1 moiety, the
ester is transformed to the desired aldehydes o~
gormula (3) using standard Dihal reduction techniques,
particularly in this situation (wherein P1 is not a residue
of a natural amino acid). Alternatively, (as is preferred
when P1 is a residue of a natural amino acid) the ester is
de-esterified to its corresponding acid, converted to its
corresponding hydroxamate and the hydroxamate upon
treatment with lithium aluminum hydride is converted to its
aldehyde. In the instance wherein Pa represents a P'2
W09G102499 ~ ~ ~ ') ~
PCTlL1S951072:~I
_25_
moiety, the ethyl ester of compounds (19) are removed and
the resulting compounds are ready for coupling as outlined
in Reaction Scheme A'.
In Scheme D an alternative procedure for preparation of
compounds of formula IA is set forth.
to
20
30
W'O 9GICI2-t99 PCT'IU995/072d1
~I'~')~rr~J
_z~_
Reaction Scheme D
(~}
( a), Coupling Reaction
P2 Pt
I 1
PgNH-CH-CNH-CH -C-CFz-C-NRSRg
I
O (23) OH O
(b), Deprotedion
P2 Pt
I
HzN-CH-CNH-CH -C-CFZ-C-NRSRs
) I
p OH O
(24)
(c}, Coupling Reaction
(d}, Oxidation
P2 Pt
R'tC-NH-CH-CNH-CH -C-CFz-C-NR5R5
O 0 IA O O
In Scheme D, step (a) compounds of formula (7) are
subjected to a coupling reaction in a manner analogous to
that described previously in Scheme A' step (e) with a
suitably protected acid of the formula PgNHCH(P2)CQ2H to
provide compound of formula (23).
In Scheme D, step (b) compounds of formula (23) are
deprotected in a manner analogous to that described in
Scheme A, step (d) to provide compounds of formula (24).
In Scheme D, steps (c) and (d) compounds of formula
(24) are first subjected to a coupling reaction with an
acid of the formula R'1C02FI in a manner analogous to that
described in Scheme A' step (2) and the coupled product is
W'O 96I(12199 2 '~ ~ ~ ~ L ~ PCTJUS9g107241
-z~-
then oxidized in a manner analogous to that described in
Scheme A' step (f) to provide the compounds of formula IA.
It is understood by one of ordinary skill in the art
that compounds of formulas (9) and (15) may be manipulated
in a manner analogous to formula (7) in Scheme D, in
addition to the manipulations decribed in Schemes A' and A"
to provide compounds of formulas IA, IA', IB and IB'.
Having generically described the methods for the
preparation of the compounds of this invention, the
following specific examples illustrate the chemistry and
techniques by which the synthesis may be effected.
The following examples present typical syntheses as
described in Schemes A, A', A" and D. These examples are
understood to be illustrative only and are nat intended to
limit the scope of the present invention in any way. As
used herein, the following terms have the indicated
meanings: "g" refers to grams; "mmol" refers to millimoles;
"ml" refers to milliliters; "bp" refers to boiling point;
"mp" refers to melting point; "'°C" refers to degrees
Celsius; "mm Hg" refers to millimeters of mercury; "uL"
refers to microliters; "up" refers to micrograms; "uM"
refers to micromolar; "Cbz" means carbobenzyloxy; "DMF"
means dimethylformamide; "THF" means tetrahydrofuran;
"TBAF" means tetrabutylammonium fluoride; "NMM" means N-
methylmorpholine; "DMSO" means dimethylsulfoxide; "HOBT"
means hydroxybenzotriazole and "EDC" means 1-(3-
dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride.
4fO96102a9!> ~ ~ C~ ~ j ~ ~ PCTI0895lfi24l
-28-
Exam~rle 1
Preparation of a,a-Difluoro-Y-[[(2-(Ry-
[[(hvdroxyyphenylacetyl]amino]-3-methyl-1-oxobutvl}amine]-
~oxo-4-(phenylmethoxy~-N-(phenylmethyl)-benzene-
.~entanamide.
OH O
1D NH~ CFZ 'NH
- HH
O O
Step A
preparation of O-benzyl-N-(tert-butoxycarbonyl3-L-
tyrosinal- the starting material in Reaction Scheme A
(Following the procedure of Schirlin, D. and Van
Dorsselaer, V. in PCT/CTS91/09741 published 3uly 23, 1992
with an international publication number of H10 92/12123.1
A mixture of N-tert-butoxycarbonyl-L-O-benzyltyrosine
(37.1 g, 100 mmol), dicyclohexylcarbodiimide (20.6 g. 100
moral), and N-hydroxybenzotriazole hydrate (15.3 g, 1DD
mmol} in anhydrous dichloromethane (35D mL) is stirred at
0°C for 10 minutes. To this is added at 0°C, N_,O_-
dimethyihydroxylamine hydrochloride (9.75 g, 100 mmol} and
~F-methylmorpholine (I0.1 g, 100 mmol). The temperature is
allowed to warm to room temperature and stirring is
continued for 15 hours. The white precipitate is then
filtered off and rinsed with dichioromethane. The filtrate
is concentrated under vacuum and the residue is purified by
Plash chromatography (silica gel, ethyl
acetate/cyclohexane, 2:8) to provide the N-tert-
butoxycarbonyl-L-0-benzyltyrosine-N, On-dimethyl-hydroxamate
(34.3 g) as a white solid (Rp=0.36 in ethyl
acetatejcyclohexane, 1:1}.
WO 96102499 J ~f ~ PCT/US95107241
-29
The N-tent-butoxycarbonyl-L-O-benzyltyrosine-N,O
dimethyl-hydroxamate (18.2 g, 44 mmol) is dissolved in a
mixture of anhydrous diethyl ether/dimethoxyethane (300 mL,
4:1) and cooled to 0°C. To this is added lithium aluminum
hydride (1.82 g, 48 mmol) portionwise. The reaction is
stirred at 0°C for 1.5 hours. A 1M solution of potassium
hydrogen sulfate (55 mL) is then added dropwise with
stirring to the reaction. After addition is complete, the
aqueous phase is decanted and extracted with ethyl acetate
(2 x 200 mL). The combined organic layers are washed with
3N hydrochloric acid (250 mL), water (200 mL), saturated
sodium bicarbonate (150 mL) and brine (200 mL). The
organic layer is then dried over anhydrous magnesium
sulfate, filtered and concentrated under vacuum. The
residue is recrystallized from ethyl acetate/gentane to
provide N-tert-butoxycarbonyl-L-O-benzyltyrosinal (13 g).
Step B
Preparation of 4-tert-butoxycarbonylamino-2,2-difluoro-3
hydroxy-5-(4-benzyloxy)phenylpentanoic acid ethyl ester.
Reaction Scheme A, step (a); To a stirred mixture of
N-tert-butoxycarbonyl-L-O-benzyltyrosinal (13.0 g, 36.6
mmol), silver acetate (1.82 g, 10.9 mmol), activated zinc
dust (5.02 g, 76.8 mg-atom, washed with 3N hydrochloric
acid, water, acetone and ether) and ethyl
bromodifluoroacetate (14.8 g, 72.9 mmol) in anhydrous
tetrahydrofuran (120 mL) at 0°C is added diethylaluminum
chloride (22.4 mL of a 1.8M solution in toluene) over 20
minutes. The temperature is kept below 12°C during the
addition. The reaction is then allowed to stir at 0°C for
90 minutes and then at room temperature for 4 hours. The
reaction is then cooled to 10°C and quenched with saturated
aqueous ammonium chloride (200 mL). A 1M solution of
sodium hydrogen tartrate (200 mL) is added and the reaction
is allowed to warm to room temperature. The reaction is
filtered and the solids rinsed with ethyl acetate. The
filtrate layers are separated and the aqueous layer is
~: ~ 5
wo vs~a~?.~vv ~- ' .~) ~ ~- rcT~~usvsro7za~
-30-
extracted with ethyl acetate. The combined organic layers
are dried over anhydrous magnesium sulfate, filtered arid
concentrated under vacuum. The residue is purified by
flash chromatography (cyclohexaneJethyl acetate, 4:1) to
provide the title compound (8.3k g'). The ratio of
diastereomers is approximately 1:1.
Step C
Preparation of 4-tert-Butoxycarbonylamino-2 2-difluoro-3
1G _hydroxv-5-(4-benzyloxv)phenyl-N-(phenvlmethyl)pentanamide.
Reaction Scheme A step (b): To a solution of 4-tert
butoxycarbonylamino-2,2-difluoro-3-hydraxy-5-(4
benzyloxy)ghenylpentanoic acid, ethyl ester (5.5 g, 11.5
mmol) in anhydrous tetrahydrofuran (50 mL) is added at 0°C,
1: benzylamine (6.15 g, 57.5 mmo1). The reaction is stirred
fer 3 hours at 0°C, then at room temperature for 15 hours.
The reaction is then diluted with ethyl acetate (100 mL),
washed with O.1N aqueous hydrochloric acid (2 x 50 mL),
water (50 mL), brine (50 mL) and dried over anhydrous
20 magnesium sulfate. It is then filtered and concentrated
under vacuum. The residue is recrystallized from ethyl
acetate/pentane to provide the title compound (5.17 g) as a
white solid.
25 Step D
Preparation of 4-amino-2,2-difluoro-3-hvdroxv-5-(4-
benzyloxy)phenyl-N-(phenylmethyl)pentanamide.
Reaction Scheme A, step (d): A solution of 4-tert-
butoxycarbonylamino-2,2-difluoro-3-hydroxy-5-(4-
30 benzyloxy)phenyl-N-(phenylmethyl)pentamide (5.1 g, 9.4
mmol) in trifluoroacetic acid (200 mL) a stirred at 0°C for
1 hour. The solvent is then removed under vacuum and the
residue dissolved in ethyl acetate (100 mL). The organic
layer solution is washed with saturated sodium hicarbonate
35 (3 x 50 mL), brine, dried over anhydrous magnesium sulfate,
filtered and concentrate under vacuum to provide the title
11'096FOta99 ~ j ~.~ J ~ ~ y PCTIiJ8951032d1
-31-
compound as a white solid; Rf=0.62 (silica gel,
butanol/acetic acid/water, 6:2:2).
Step E
Preparation of N-(R)- [ (1,1-
Dimethylethyl)dimethylsilyl]oxvinhenylacetyl]-L-valine.
To a stirred solution of (R}-(-)-mandelic acid (5.6g, 37
mmol) and tent-butylchlorodimethylsilane (15.0 g, 99.5
mmol) in DMF (100 mL) is added imidazole (16.7 g, 245 mmol)
in portions over 15 minutes while maintaining the reaction
mixture near 25°C with a coal water bath. After 17 hours,
the mixture is diluted with water and extracted with two
portions of ether. The combined extracts are cooled to -
10°C and washed quickly with cold (-10°C) 1 N HC1, twice
with cold water, dried over anhydrous sodium sulfate,
filtered and concentrated under vacuum to provide 15.0 g of
a yellow oil which is then dissolved in CH30H (250 mL) and
THF (80 mL). A solution of potassium carbonate (14 g, 100
mmol) in water (140 mL) is added with vigorous stirring.
2G After 1 hour, the mixture is partially concentrated under
vacuum and the remaining clear light yellow solution is
diluted with brine (300 mL). The cloudy mixture is cooled
to -10°C and acidified to pH 5 with 1 M KHSOq. Extraction
with cold (-10°C) ether and washing of the extracts with two
gortions of brine provides, after drying with anhydrous
sodium sulfate, filtration and concentration under vacuum,
6.6 g (678) of (R)-O-tart-butyldimethyl-silylmandelic acid
as a yellow oil. This crude silylated acid (25 mmol) is
dissolved in CH3CN (80 mL) and the resulting solution is
3G cooled to -10°C. To the stirred solution is added NMM (2.52
g, 24.9 mmol), then isobutylchloroformate (3.72 g, 27.2
mmol). After 10 minutes, a solution of L-valine methyl
ester hydrochloride (7.32 g, 43.7 mmol) and NMM (4.08 g,
40.3 mmol) in HZO (20 mL) is added and stirring is continued
at -10°C for 20 minutes. The reaction mixture is poured
into ice-cold dilute HC1 containing some NaCl and extracted
with two portions of ethyl acetate. The combined extracts
!f0 9tiltJr~99 ~ ~ ~ ~ ) ~ .~ PCTlUS9510724!
-32-
are washed with water, aqueous saturated sodium
bicarbonate, brine, dried over anhydrous magnesium sulfate,
filtered and concentrated under vacuum. Two flash
chromatographies (12% ethyl acetate in cyclohexane) provide
4.4 g of coupled silylated ester (Rp 0.4, 4/1.
cyclohexane/ethyl acetate) contaminated with N-
isobutyloxycarbanyl-L-valine methyl ester. The silylated
ester (4.4 g, 12 mmol) is dissolved in THF (15 mL) with
stirring and a solution of 1 M tetrabutylammonium fluoride
(TBAF) in THF (11.6 mL) is added. After 1.25 hours, the
solution is concentrated under vacuum and the residue is
purified by flash chromatography (4/1 cyclohexanejethyl
acetate) to provide 2.4 g (37% overall from O-tert-
butyidimethylsilylmandelic acid) of N-{(R)-mandeloyl]-L-
valine methyl ester as an oil. Following the procedures in
a manner analogous to that described above, the material is
reconverted to its O-tert-butyldimethylsilyl ether in 82%
yield after flash chromatography (12% ethyl acetate in
cyclohexane). To a stirred solution of this ester (13.5
mmol) in CH30H (125 mL) and water (5 mL) is added LiOH~H20
(0.S9 g, 14 mmol). After 17 hours an additional amount of
LiOH.H20 (0.06 g) is added and stirring is continued for 2
hours. The solution is concentrated under vacuum and the
residue is diluted with water (100 mL) and washed with
ether. The aqueous layer is acidified with ice-cold 6 N
HC1 and extracted with two portions of ether. The combined
extracts are washed with water, dried over anhydrous
magnesium sulfate, filtered and concentrated under vacuum
to provide the title compound in 48% yield as a white
solid: mp 146-148°C; IR, (CHC13) vma% 2957, 2932, 1718,
1678, 1516, 1096, 863, 840 cm-r; 1H N'MR (CDC13) d 9.17 (bs,
1 H), 7.51 (d, 1 H, J=9.0 Hz), 7.43-7.40 (m, 2 H), 7.32-
7.25 (m, 3 H}, 5.15 (s, 1 H), 4.51 (dd, 1 H, J=9.0, 4.3
Hz), 2.34-2.28 (m, 1 H), 1.00 (d, 3 H, J=6.9 Hz), 0.97 (d,
3 H, J=7.0 Hz), 0.94 (S, 9 H), O.I1 (S, 3 H), -0.08 (5, 3
H); 13C NMR (CDC13) d 175.49, 172.65, 139.36, 128.32,
128.21, 126.68, 75.66, 56.6D, 3i. D4, 25.66, 19.0I, 18.02,
WO 96l(12~99 ~ ~ ~ ~ ~ ~ ~ PCT/US95107241
-33-
17.44, -4.83, -5.31; mass spectrum, m/z 394 (M+ + 29), 366
(M+ + 1, 100), 350, 308; exact mass calcd for C19H32NOqSi
366.2101, found 366.2099; Cal2oD -56.6° (c 1.00, CH30H).
Anal. Calcd for CIyH31N04Si: C, 62.43; H, 8.55; N, 3.83.
Found: C, 62.22; H. 8.61; N, 3.65.
Step F
Preparation of [3~,4(5,R)]-2,4 5-Trideoxy-4-[ 2- [([(1 1-
dimethylethyl)dimethylsilyl)oxy]phenvlacetyl]amino)-3-
methyl-1-oxobutyl]amino]-2,2-difluoro-5- 4-
(phenylmethoxy)phenyll-N-(phenylmethyl)-L-alycero-
pentanamide.
Reaction Scheme A', step (e): N-(R)-[[[(I,1
dimethylethyl)dimethylsilyl]oxy]phenylacetyl)-L-valine
prepared above (25 mmol) is dissolved in CHgCN (80mL) and
the resulting solution cooled to -10°C. To the stirred
solution is added N-methylmorpholine (2.52 g, 24.9 mmol,
NMM), followed by isobutylchloroformate (3.72 g, 27.2
mmol). After 10 minutes, a solution of 4-amino-2,2-
difluoro-3-hydroxy-5-(4-benzyloxy)phenyl-N-(phenylmethyl)
pentanamide (43.7 mmol, prepared in step (d) above) and NMM
(4.08 g, 40.3 mmol) in H20 (20 mL) is added and stirring is
continued at -10°C for 20 minutes. The reaction mixture is
poured into ice-cold dilute HC1 containing some NaCl and is
extracted with two portions of ethyl acetate. The combined
extracts are washed with water, aqueous saturated sodium
bicarbonate, brine, dried over anhydrous magnesium sulfate,
filtered and concentrated under vacuum. The title compound
is obtained in 768 yield as a clear colorless oil which
crystallizes on standing after flash chromatography (68
acetone in CH2ClZ): mp 117-120°C; IR (KBr) vmax 3418, 1704,
1677, 1653, 1540, 1513, 1246, 1095, 839 cm-1; 1H NMR (CDC13)
d 7.44-7.18 (m, 16 H), 6.96 (d, 3 H, J=8.7 Hz), 6.84 (d, 2
H, J=8.7 Hz), 6.10 (d, 1 H, J=8.1 Hz), 5.10 (s, 1 H), 5.01
(s, 2 H), 4.69 (d, 1 H, J=6.9 Hz), 4.43 (dd, 1 H, J=14.7,
6.0 Hz), 4.34 (dd, 1 H, J=14.7, 6.0 Hz), 3.96 (dd, 1 H,
3=8.5, 6.9 Hz), 3.91-3.82 (m, 2 H), 2.76 (dd, 1 H, J=13.8,
«
'O 9ti102-t99 ~ ~ ~ '~ ~ ~ 5 Pf.'T!tiS951072di
-34-
7.8 Hz}, 2.66 (dd, 1 H, J=13.8, 7.8 Hz), 2.22-2.11 (m, 1
H), 0.94 (s, 9 H), 0.88 (d, 3 H, J=b.6 Hz), 0.86 (d, 3 H,
J=6.9 Hz). 0.09 (s, 3 H), -U.06 (s, 3 H); 13C NMR (CDCi3) d
173.37, 171.91, 163.92 (t, J=35 Hz), 159.01, 139.62.
137.40, 137.31, 130.47, 129.60, 129.16, 128.94, 128.80,
128.70, 128.37, 128.31, 128.22, 127.83, 126.63, 115.50 (t,
J=257 Hz), 115.36, 76.21, 70.67 (t, J=25 Hz), 70.37, 58.79,
51.93, 43.76, 36.52, 29.97. 26.D8, 19.80. 18.47, 17.96, -
4.41, -4.89; 19F NMR (CDC13) d -118.40 (s), -118.45 (s);
mass spectrum, m,'z 816 (M+ + 29). 788 (M+ + 1), 573, 469,
441, 423 (100); exact mass calcd for CqqHS6FZN3QSSi 788.3906,
found 788.3932: (a]z°D -74.6° (c 1.02, CH30H). Anal. Calcd
for CqqHSSFZN306Si: C, 67.06; H, 7.04; N, 5.33. Found: C,
67.07; H, 7.14; N, 5.18.
1J
Step G
Preparation of final title compound.
Reaction Scheme A', steps (f) and (g): The above
prepared alcohol is oxidized under Swern conditions as
2ii follows: To a stirred solution of 2 M axalyl chlaride/CHyClz
(2.OmL) at -60°C under nitrogen is added dropwise anhydrous
DMSO (0.42 mL, 5.9 mmol). After 10 minutes, a solution of
[3~,4(S,R)]-2,4,5-Trideoxy-4-[[2-[(([(1,1-
dimethylethyl}dimethylsilyl]oxy]phenyiacetyl]amino]-3-
25 methyl-1-oxobutyl]amino]-f,2-difluoro-5-[4-
(phenylmethoxy)phenyl]-N-jphenylmethyl)-L-glycero-
pentonamide (0.50 mmol) in CHZClz (3 mL) and anhydrous DMSO
(1.5 mL) is added. The solution is allowed to stir at -65
to -55°C for 5 hours. It is then warmed to -20°C, followed
30 by addition of triethylamine (1.39 mL, 10.0 mmol) over 5
minutes. The reaction mixture is then allowed to warm to
room temgerature overnight. it is then diluted with ethyl
acetate (60 mL) and filtered, washing the solids with ethyl
acetate. The filtrate is washed with 0.1 M HC1 (3 x 15
3~ mL), brine, dried over anhydrous magnesium sulfate,
filtered and concentrated under vacuum to provide 375 mg of
crude ketane. This is dissolved in 4/1 acetic acid,~HZO (5U
WO 9G1(i2.199 ~ PCTIU5951072a1
-35-
mL). The stirred solution is heated at 80°C for 19 hours,
allowed to cool to roam temperature before being diluted
with water and extracted with two portions of CH2Clz. The
combined extracts are washed with water, brine, dried over
anhydrous magnesium sulfate, filtered and concentrated
under vacuum. Flash chromatography of the residue (10~
acetone in CH2Clz} provides 180 mg title compound
contaminated by residual silylated compound. The material
is resubjected to the reaction conditions and purified by
flash chromatography to provide 121 mg of final title
compound. Two recrystallizations from ethyl
acetate/pentane provide 30 mg (8.8$) of final title
compound as a pale yellow solid: mg 144-150°C; IR (KBr) v,~ax
3398, 33D0, 1683, 1656, 1540, 1513, 1454, 1244, 698 cm 1;
1H NMR (DMSO-ds} d 9.77-9.72 (m, 1 H), 8.69 (d, 1 H, J=7.6
Hz), 7.80 and 7.73 (2d, 1 H total, J=9.3 Hz), 7.45-7.21 (m,
15 H}, 7.12 and 7.10 (2d, 2 H total, J=8.7 Hz), 6.89 and
6.87 (2d, 2 H total, J=8.8. 8.4 Hz), 6.21 and 6.20 (2d, 1 H
total, J=5.1 Hz), 5.05 (s, 2 H). 5.02 and 5.00 (2s, 1 H
total), 5.03-4.9D (m, 1 H), 4.38 (d, 2 H, J=5.7 Hz}, 4.23-
4.17 (m, 1 H}, 3.14-3.07 and 2.65-2.57 (m, 2 H total),
1.97-1.88 and 1.78-1.73 (m, 1 H total), 0.73 and 0.71 (2d,
3 H total, J=6.7 Hz), 0.57 and 0.49 (2d, 3 H total, J=6.9.
6.7 Hz); 19F NMR (DMSO-db) 8 -110.31 (d, J=268 Hz), -110.54
(d, J=268 Hz), -112.21 (d, J=268 Hz), -112.25 (d, J=269
Hz); mass spectrum, m/z 700 (M+ + 29), 672 (M+ + I, 100),
652, 439, 421, 402; exact mass calcd for C3gH4pFZNgOs
672.2885, found 672.2842.
35
wo ~c~ni2;~as ~ ~ q e: t :' ~j PcTtus~3sro~aay
~~~._
-36-
Example 2
Preparation of [6S-(6R*, 9R*, 13S*)]-4,4,-Difluaro-9-
methvlethvl)-3.5,8.11-tetraoxo-1,13-diahenvl-6-ff4-
(phenylmethoxy)phenyl]methyl]-12-oxa-2,7,10-
triazatetradecan-14-oic Acid, Methyl Ester.
r /
'' O H~ CF2~~1H
1/ ~ ~~lJJ(H
CH302C O ~ O O
Step A
Preparation of [3~,4(S)]-2,4,5-Trideoxy-4-[[2-[j~l,l-
dimethylettaoxy)carbonyl]amino]-3-methyl-1-oxobutyl]amino-S-
[4-(phenylmethoxy)phenyl]-N-(phenylmethyl)-L-4lycero-
pentanamide.
Reaction Scheme D, step (a): To an ice-cold stirred
solution of 4-amino-2,2-difluoro-3-hydroxy-5-(4-
benzyloxy)phenyl-N-(phenylmethyl)pentanamide (556 mg, 1.23
ms~ol, prepared in example i, step D) in 2:1 dry CHZCl2/DMF
(13.5 mL) is added BOBT (0.19 g, 1.2 mmol), DCC (0.268, 1.3
mmalj and N-t-Boc-L-valine (0.27 g, 1.2 mmol). The
reaction mixture is allowed to warm to 25°C overnight, then
2S
diluted with ethyl acetate/cyclohexane, washed with dilute
aqueous sodium bicarbonate, water, brine, dried over
anhydrous magnesium sulfate, filtered and concentrated
under vacuum to provide a gummy solid. This is triturated
with CHpCl2 and filtered to remove some of the
dicyclohexylurea. The filtrate is concentrated under
vacuum and the residue is purified by flash chromatography
(3:2 cyclohexane/ethyl acetate) to provide 694 mg (88~j of
the title compound. as an ivory solid. The major
3S diastereomer is separated in 69~ yield by recrystallization
from etherfCH2C12.
WO 967(12499 ~ ~ (~ C~ °~ ~ ~ PCTILiS95/07241
s
-37-
Step B
Preparation of the chloroformate of formula:
/ O»CI
CH302C ~~'~(O
To a stirred solution of triphosgene (6.65 g, 22.4
mmol) in CH2C12 (25 mL) under nitrogen is added (R)-(-)-
methyl mandelate (5.36 g. 32.3 mmol) followed by the rapid
addition of a solution of pyridine (2.66 mL, 32.9 mmol) in
CH2C12 (2.5 mL). The reaction mixture is warmed to reflux
and pyridinium chloride separated. After 17 hours at room
temperature, the reaction mixture is partially concentrated
under vacuum, diluted with ethyl acetate, and filtered.
The filtrate is concentrated concentrated under vacuum and
the residue is again dissolved in ethyl acetate and
filtered. Concentration under vacuum provides 7.25 g of a
pale yellow oil composed of approximately a 20:1:1 mixture
of chloroformate: carbonate: methyl mandelate. For the
chloroformate IR (neat) vmax 1772. 1750 cm-1; 1H NMR (CDClg)
d 7.5-7.37 (m, 5 H). 5.98 (s, 1 H), 3.78 (s, 3 H).
Step C
Preparation of [SE.6S-(6R*, 9R*, 135*)]-4,4,-Difluoro-5-
hydroxy-9-(1-methylethyl)-3,8,11-trioxo-1,13-diphenyl-6-
[[4-(phenylmethoxy)phenyl]methyl]-12-oxa-2,7.10-
triazatetradecan-14-oic Acid, Methyl Ester.
Reaction Scheme D, steps (b) and (c): A flask
containing the major diastereomer of (3g,4(S)]-2,4,5-
Trideoxy-4-[[2-[[(1,1-dimethylethoxy)carbonyl]amino]-3-
methyl-1-oxobutyl]amino-5-[4-(phenylmethoxy)phenyl]-N-
(phenylmethyl)-L-glycero-pentonamide (403 mg, 0.630 mmol)
is submersed in an ice bath and ice-cold trifluoroacetic
acid (TFA) (5 mL) is added with stirring. After 30
minutes, the solution is concentrated under vacuum at 25°C
and the residue is carefully partitioned between ethyl
acetate and aqueous saturated sodium bicarbonate. The
R'O 9Gt0:-t99 ~ ~ ~ j ~ ,- PCTIU595I0724I
L ~ ~,.~ d 4y~
-3$-
organic layer is separated, washed with brine, dried over
anhydrous magnesium sulfate, filtered and concentrated
under vacuum to provide 334 mg (98%) of deprotected amine
which is immediately suspended in CHZClz (5 mL) at 0°C and
treated with NMM (77 uL. 0.70 mmol) and the chloroformate
prepared above (160 mg, 0.70 mmol) dissolved in CHZCl2 (1
mL). After stirring for 1.5 hours, the reaction mixture is
poured into ethyl acetate/dilute aqueous ammonium chloride.
The organic layer is separated, washed with dilute aqueous
1G sodium bicarbonate, brine, dried over anhydrous magnesium
sulfate, filtered and concentrated under vacuum. Flash
chromatography (4:1 CHZCIZ/ethyl acetate) of the residue
provides 332 mg (72%) of a single diastereomer o~ the title
compound as a white foam: 1H NMR (CDC13) $ 7.44-7.26 (m, 15
H), 7.1.2 (d, 2 H, J=8.4 Hz), 6.89 (d, 2 H, J=8.4 Hz), 6.53
(d, 1 H, J=8.4 Hz), 5.89 (s. 1 H), 5.40 (d, 1 H, J=7.9 Hz),
5.02 (s, 2 H), 4.70 (d, 1 H, J=7.0 Hz), 4.49 (apparent d, 2
H, J=5.8 Hz), 4.3-4.1 (m, 2 H), 3.85 (dd, 1 H, J=7.8, 5.5
Hz), 3.63 (s, 3 H), 3.05-2.87 (m, 2 H), 2.2-2.04 (m, 1 H),
1.90 (s, 1 H), 0.83 (d, 3 H, J=6.7 Hz), 0~71 (d, 3 H, J=6.8
Hz); 1gF NMR (CDC13) d -116.60 (dd, 1 F, J=259, 9 Hz), -
118.73 (dd, 1 F, J=259, 15 Hz),
Step D
23 _Pr_eparation o~ final title compound.
Reaction Scheme A', step (f): The above prepared
alcohol is subjected to Swern Oxidation conditions in a
manner analogous to that described previously in example I,
step G. The residue is purified by flash chromatography
(4:1. CHZC12/ethyl acetate) to provide 52% of the final title
compound. Precipitation with pentane from an ethyl acetate
solution provides 29% of final title compound as a white
powder: IR (KBr) vm8x 3399, 3316. 1745, 1699, 1668, 1534,
1513, 1241 cm-1; IH NMR (CDC13) d 5.89, 5.87, and 5.82 (3s
in 6.0:1.$:1.0 ratio, 1 H); 5.05 and 5.03 (2s in 1:1.6
ratio, 2 H); 3.69. 3.67, and 3.65 (3s in 2.5:1.0:1.9 ratio,
3 H); 19F NMR (CDCI3) 8 -112.53 (s), -112.60 (s), -116.25
W096102-t99 L ~ ~ ~) ~ ~ ~ PCTlUS95107241
-39-
(d, J=256 Hz), -117.00 (d, J=256 Hz), -12D.21 (d, J=256
Hz), -120.78 (d, J=256 Hz), (s: d ratio 4:1); mass
spectrum, m/z 770 (M+ + 41), 758 (M+ + 29), 731, 730 (M+ +
1, 100); exact mass calcd for CqpHq2F2N30g 730.2940, found
730.2925; [a]2°D -48.3° (c 1.22, CH30H). Anal. Calcd for
CqpHq1F2N30g: C, 65.83; H, 5.66; N, 5.76. Found: C, 65.16;
H, 5.69; N, 5.79.
Example 3
Preparation of a,a-Difluoro-y-([3-methyl-1-oxa-2-[[(1-
oxo-traps-3-phenyl-2-propenyl)amino]butyl]amino]-B-oxo-4-
(phenylmethoxy)-N-(phenvlmethyl)-benzene-pentanamide.
n
O
IH
Step A
Preparation of N-(traps-Cinnamoyl)-L-valine
To a stirred solution of L-valine methyl ester
hydrochloride (1.97 g, 11.8 mural) and NMM (1.10 g, 10.8
mmol) in CH2C12 (15 mL) and DMF (20 mL} is added a solution
of 1-traps-cinnamoylimidazole (2.15 g, 10.8 mmol) in CH2C12
(28 mL). After 18 h, additional L-valine methyl ester
hydrochloride (0.36 g, 2.1 mmol) is added and the reaction
3D mixture is stirred for 5 h before being diluted with water
(100 mL) and extracted with CHC13. The organic extract is
washed with three portions of water, dried (MgSOq), and
concentrated in vacuo. Filtration through a short silica
gel column (1/1 ethyl acetateJcyclohexane) provides 2.0 g
(71~) of N-(traps-cinnamoyl)-L-valine methyl ester. To a
stirred solution of this ester (13.5 mmol) in CH30H (125
mL) and water (5 mL) is added LiOH~H20 (0.59 g. 14 mmol).
After 17 hours an additional amount of LiOH~H20 (0.06 g) is
~y091i102a99 ~ ~ ~ ~ ~ ~ °~ P('TIU895f072.~1
-40-
added and stirring is continued for 2 hours. The solution
is concentrated in vacua and the residue is diluted with
water (100 mL) and washed with ether. The aqueous layer is
acidified with ice-cold 6 N HC1 and extracted with two
portions of ether. The combined extracts are washed with
water and dried (MgS04). Concentration in vacua provides
67~ of the title compound as a white solid: mp 63-68°C; IR
(CHC13) v",gx 3008, 2970, 1718, 1670, 1629, 1512, 1196, 667
cm-1; 1H NMR (CDC13) d 9.94 (bs, 1 H), 7.65 (d, 1 H, J=15.6
1D Hz), 7.50-7.46 (m, 2 H), 7.35-7.31 (m, 3 H), 6.56 (d, 1 H,
J=8.7 Hz), 6.52 (d, 1 H, J=15.6 Hz), 4.75 (dd, i H, J=8.7,
4.8 Hz), 2.34-2.28 (m, 1 H), 1.02 (d, 3 H, J=6.9 Hz), 0.99
(d, 3 H, J=6.6 Hz); 13C NMR (CDC13) d 175.31, 166.66,
142.42, 134.50, 129.93, 128.79, 127.94, 119.75, 57.47,
31.19, 19.00, 17.78; mass spectrum, mjz 276 (&I* + 29), 248
(M* + 1), 203, 131 (100); exact mass calcd far ClmH1gN03
248.1287, found 248.1290; [a]20p +24.1° {c i.02, CH30H).
Anal. Calad for CIqHI~NOg~O.1H20: C, 67.51; H, 6.96; N,
5.62. Found: C, 67.30; H, 7.10; N, 5.57.
Step B
Preparation of (3~ 4(S)]-2 4 5-Trideoxv-2,2-difluoro-4-[j3-,
metal--1-axo-2-{C1-oxo-traps-3-phenyl-2-
prapenyl)am'no]butyl]amino)-5-[4-(phenylme'hoxy)phenyl]-N-
(phenylmethvl)-L-alycero-pentanamide.
Reaction Scheme A', step (e): N-{traps-Cinnamoyl)-L-
valine prepared above {25 mmol) is dissolved in CH3CN (80mL)
and the resulting solution cooled to -10°C. To the stirred
solution is added N-methylmorpholine (2.52 g, 24.9 mmol,
NMdi), followed by isabutylchloroformate (3.72 g, 27.2
mmol). After 10 minutes, a solution of 4-amino-2,2-
difluoro-3-hydroxy-5-{4-benzyloxy)phenyl-N-(phenylmethyl)
pentanamide (43.7 mmol, prepared in examgle 7., step D and
NH1M {4.08 g, 40.3 mmoi) in HZO (20 mL) is added and stirring
is continued at -10°C for 20 minutes. The reaction mixture
is poured into ice-cold dilute HC1 containing some NaCl and
is extracted with two portions of ethyl acetate. The
~ ~ " "lC
WO 9G!02499 ~ ~ ~ '~ 1 ~ ') PCTIUS95/07241
-41-
combined extracts are washed with water, aqueous saturated
sodium bicarbonate, brine, dried over anhydrous magnesium
sulfate, filtered and concentrated under vacuum. The
residue is recrystallized from ethyl acetate/pentane to
provide 61~ of a 2:1 mixture of diastereomers of title
- compound as a light tan solid: mp 222-227°C; IR (KBr) vmax
3410, 3287, 1682, 1652, 1620, 1540, 1512, 1243, 1217, 697
c:a 1; 1H NMR (DMSO-db) 8 9.17-9.11 (m, 1 H), 8.05 and 8.01
(2d, 1 H total, J=9.1, 9.1 Hz), 7.85 and 7.71 (2d, 1 H
total, J=9.1, 9.0 Hz), 7.59-7.22 (m, 16 H), 7.12 (d, 2 H,
J=8.5 Hz), 6.94-6.85 (m, 3 H), 6.26 (d, 1 H, J=7.6 Hz),
5.04 and 4.96 (2s, 2 H total), 4.42-3.99 (m, 5 H), 2.79-
2.63 (m, 2 H), 2.09-1.98 and 1.92-1.85 (2m, 1 H total),
0.86 and 0.74 (2d, 3 H total, J=6.9 Hz), 0.83 and 0.64 (2d,
3 H total, J=6.9 Hz); 19F NMR (DMSO-dg) E -111.11 (dd,
J=255, 6 Hz), -112.12 (dd, J=254, 8 Hz), -120.35 (dd,
J=254, 19 Hz), -122.04 (dd, J=255, 20 Hz); mass spectrum,
m/z 698 (M+ + 29j, 670 (M+ + 1, 100), 650, 441, 423. Anal.
Calcd for C39H41F2N30S: C, 69.94; H, 6.17; N, 6.27. Found:
C, 68.59; H, 6.32; N, 5.90.
Step C
Preparation of the final title compound.
Reaction Scheme A', step (f): The above prepared
alcohol is oxidized under Swern conditions as follows: To
a stirred solution of 2 M oxalyl chloride/CH2C12 (2.OmL) at
-60°C under nitrogen is added dropwise anhydrous DMSO (0.42
mL, 5.9 mmol). After 10 minutes, a solution o~ the above
prepared alcohol (0.50 mmol) in CH2C12 (3 mL) and anhydrous
DMSO (1.5 mL) is added. The solution is allowed to stir at
-65 to -55°C for 5 hours. It is then warmed to -20°C,
followed by addition of 'triethylamine (1.39 mL, 10.0 mmol)
over 5 minutes. The reaction mixture is then allowed to
warm to room temperature overnight. It is then diluted
with ethyl acetate (60 mL) and filtered, washing the solids
with ethyl acetate. The filtrate is washed with 0.1 M HC1
(3 x 15 mL), brine, dried over anhydrous magnesium sulfate,
~~0 9s~oz.vs rc~ra~s~smaai
~°~ ~~~ LJ
-42-
filtered and concentrated under vacuum. Flash
chromatography (6~ acetone in CHxCl2) followed by
recrystallization from ethyl acetate/pentane provides 34~;
of the final title compound as a white solid: mp 187-194°C;
IR (KBr) vm$X 3403r 3285, 1691, 1654, 1623, 1540, 1513,
1454, 1243, 1219, 1178, 697 cm-1; 1H NMR (DMSb-d6) d 9.77-
9.68 (m, 1 H), 8.fi7-8.62 (m 1 H), 8.05-8.00 (m, 1 H), 7.56
(d, 2 H, J=7.0 Hz), 7.50-7.21 (m, 14 H), 7.14 (d, 2 H,
J=8.5 Hz), 6.91-6.83 (m, 3 H), 5.04-4.95 (m, 3 H), 4.42-
1Q 4.33 (m, 3 H), 3.14-3.05 (m, 1 H), 2.74-2.61 (m, 1 H),
2.02-1.76 (m, 1 H), D.84 and 0.81 (2d, 3 H total, J=7.0,
6.9 Hz), 0.64 and 0.59 (2d, 3 H total, J=7.1, 6.9 Hz); 1gF
NMR (DMSO-dg) d -110.26 (d, J=269 Hz), -110.53 (d, J=268
Hz),-112.03 (d, J=269 Hz), -112.04 (d, J=268 Hz)-major
15 diastereomer pair; -105.07 (d, J=265 Hz), -105.08 (d,
J=266 Hz), -110.59 (d, J=265 Hz), -110.61 (d, J=266 Hz)-
minor diastereomer pair; mass spectrum, mJz 696 (M+ + 29),
668 (M+ + 1), 439, 230, 202, 131, 124, 91 (100); exact mass
calcd for C3gH~pF2N305 668.2936, found 668.2953; (a)a0D
2G +7.23° (c 0.622, DMSO-db). Anal. Calcd ~or C3gH~~FZN305: C,
70.15; H, 5.89: N, 6.29. Found: C, 69.63; H, 5.85; N,
6.01.
30
1V0 96!(12199 ~ j ~~ J ~ rL '~ PCTlUS9i1072.11
-43-
Example 4
Preparation of a,a-Difluoro-Y-[[2-[2-hydroxy-1-oxo-4-
(phenylbutyl)amino]-3-methyl-1-oxobutyl]amino]-S-oxo-4-
( henylmethoxyl-N-(pheny!methyl)-benzene-pentanamide.
0
!H~ CF2 'NH
~I ((H
l0
0 O
Step A
Preparation of(R)-[[2-[(1,1-
Dimethylethyl)dimethylsilyl]oxy]-4-phenylbutyryll-L-valine.
_5
(R)-(-)-2-hydroxy-4-phenylbutyric acid is coupled with
L-valine methyl ester hydrochloride under standard coupling
conditions in a manner analogous to that described
previously, such as example 2, step A, to provide (R)-(2-
20 hydroxy-4-phenylbutyryl)-L-valine methyl ester as a yellow
solid in 89~ yield. Treatment with tert-butyldimethylsilyl
chloride in a manner analogous to that described previously
in example 1 for the preparation of N-(R)-[[[(1,1-
dimethylethyl)dimethylsilyl]oxy]phenylacetyl]-L-valine
provides the ester of the title compound as a colorless oil
in 83~ yield after flash chromatography (9/1
cyclohexane/ethyl acetate). The ester is hydrolyzed with
LiOH.H20 in a manner analogous to that described previously
in example 1 for the preparation of N-(R)-[[[(l,l-
dimethylethyl)dimethylsilyl]oxy]phenylacetyl]-L-valine to
provide the title compound in 58~ yield as a white solid:
mp 110-113°C; IR (KBr) vmax 3387, 2958, 2932, 2897. 2860,
1719, 1626, 1531, 1251, 1097, 839 cm'1; 1H NMR (CDC13) d
7.24-7.12 (m, 7 H), 4.54 (dd, 1 H, J=8.7, 4.5 Hz), 4.32 (t,
1-H, J=4.8 Hz), 2.76-2.55 (m, 2 H), 2.36-2.25 (m, 1 H),
2.19-2.07 (m, 1 H}, 2.02-1.90 (m, 1 H), 1.00 (d, 3 H, J=7.2
Hz), 0.97 (d, 3 H J=6.9 Hz}, 0.97 (s, 9 H), 0.13 (s, 3 H),
0.10 (s, 3 H); 13C NMR (CDC13) d 175.64, 174.08, 141.69,
wa vgrnza~y ~ ~ ~ ~ ~ ~ ~ ~r.Trus~~ro~2m
-44-
128.42, 128.30, 125.77, 72.73, 59.64, 36.98, 30.75, 30.02.
25.67, 19. OS, 17.93, 17.54, -4.79. -5.15; mass spectrum,
m/z 394 (M+ + 1), 393 (N+), 378, 336 (100), 289, 117, 91,
73; [a]ZED +13.7° is 1.00, CH30H). Anal. Calcd for
Ca1H35N~aSi: C, 64.08; H, 8.96; N, 3.56. Found: C, 63.87;
H, 9.07; N, 3.45.
Step B
P_r_eparation of 3~,4(S R)1-2,4 5-Trideoxy-4-[j2-[[2-[((i,1-
dimethylethyl)dimethylsilyl]oxyl-i-axo-4-
_phenyibutyl]amino]-3-methyl-1-oxobutyl]amino]-5-[4-
(phenvlmethoxv)phenvll-N-(phe~ylmethvl)-L-ctlvcero-
pentanamide.
Reaction Scheme A', step (e): (R)-[j2-[(1,1-
Dimethylethyl)dimethylsilyl]oxy]-4-phenylbutyryl]-L-vaiine
is coupled with 4-amino-2,2-difluoro-3-hydraxy-5-(4-
benzylaxy)phenyl-N-(phenylmethyl)pentanamide prepared in
example 1, step D under standard coupling conditions in a
manner analogous to that described previously, such as in
example 2, step A, to provide the title compound after
flash chromatography (6~ acetone in CHZC12) as a tan foam in
61~ yield: mp 62-67°C; 1H NMR (CDC13) d 7.44-7.11 (m, 17 H),
"7.04 (d, 2 H, J=8.7 Hz), 6.86 (d, 2 H, J=8.7 Hz), 6.11 (d,
1 H, J=8.i Hz), 5.01 (s, 2 H), 4.71 (dv 1 H, J=6.9 AZ),
23 4.52 (dd, 1 H, J=14.6, 6.5 Hz), 4.35 (dd, 1 H, J=14.4. 5.4
Hz), 4.28 (t, 1 H, J=4.8 Hz), 4.15-3.93 (m, 3 H), 2.91 (d,
2 H, J=7.8 Hz), 2.72 (ddd, 1 H, J=I3.2, 12.0, 4.8 Hz), 2.57
(ddd, 1 H, J=13.5, 12.3, 5.4 Hz), 2.15-1.90 (m, 3 H), 0.98
(S, 9 H), 0.87 (d, 3 H, J=2.4 Hz), 0.85 (d, 3 H, J=2.1 Hz),
3G 0.14 (s, 3 H), 0.10 (s, 3 H); 1gF NMR (CDC13) d -117.17 (dd,
J=259, 15 Hz), -118.62 (dd, J=259, 10 Hz); mass spectrum,
m/z 844 (M+ + 29), 816 (M+ + 1), 758, 469, 441, 423 (100),
91: [a]Z~ti -30.6° (c 0.547, CH3aH). Anal. Calcd far
CqgH5gF2NgDg5i: C, 67.70; H, 7.29; N, 5.15. Found: C, 67.32.
35 H, 7.22; N, S.1S.
wo 9s~ozasy ~ ~ '~ ~ i L ~ rcTmsysronm
-45-
Step C
Preparation of final title compound.
Reaction Scheme A', steps (f) and (g): The above
prepared alcohol is oxidized under Swern conditions in a
manner analogous to that described previously in example 1,
step G to provide the protected ketone as a light brown oil
in 72$ yield after flash chromatography (6~ acetone in
CHgClz). Deprotection of the ketone (217mg) in a manner
analogous to that described previously in example 1, step G
provides the title compound after flash chromatography (6,
10, and finally 15$ acetone in CHZC12), 104 mg (58~) as a
light yellow glass. Crystallization from CH2C12/ether and
three recrystallizations from ethyl acetate/pentane
provides 34 mg (19~) of title compound as a white solid: SR
(KBr) v",ax 3394, 3300, 1686, 1651, 1534, 1513, 1498, 1243,
698 cm-1; 1H NMR (DMSO-dfi) d 9.76-9.69 (m, 1 H) 8.70-8.66
(m, 1 H), 7.52-7.11 (m, 19 H), 6.88 and 6.87 (2d, 2 H
total, J=8.7 Hz), 5.73 (t, 1 H, J=5.8 Hz), 5.03 and 5.00
(2s overlapping m, 3H total, J=8.3 Hz), 4.35 (apparent d, 2
H, J=6.1 Hz), 4.26-4.20 (m, 1 H), 3.94-3.87 (m, 1 H), 3.15-
3.06 (m, 1 H), 2.73-2.58 (m, 2 H), 1.96-1.6fi (m, 3 H), 0.77
and 0.76 (2d, 3 H total, J=6.7 Hzy, 0.63 and 0.53 (2dr 3 H
total, J=6.8, 6.6 Hz); 19F NMR (DMSO-dg) d -110.35 (d, J=268
Hz), -110.47 (d, J=268 Hz), -112.14 (d, J=268 Hz), -112.20
(d, J=268 Hz); mass spectrum, m/z 700 (M+ + 1), 680, 622,
439 (10D), 421, 254, 91.~
35
WO 9Gf0:~~J9 " (~ ' ) g PCTIUS95IQ72d1
C l '~ ~ ~ ~. .)
-46-
Example 5
_PreDaration of N-[1-([ 3 3-Difluoro-4-([2-methyl-1-
j-{phenvlmethoxy)methyl]-prcpyl]amino]-2,4-dioxo-1-[[4-
iohenylmethoxy)phenyl]-methyllbutyllamino]carbonwl]-2-
methvlpropwl]-8-oxo-4-morpholinepropanamide.
O~ O
~N NH~ ~ CF H
NH ~ OBn
O O ~ O O
Stag A
Preparation of [1 (R) 3~ 4(S)]-2,4,5-Trideoxy-4-L[(1,1-
dimethylethoxy)-carbonyl)aminol-2 2-difluoro-N-[2-methyl-1-
L(phenylmethoxy)methyl]propel]-5-(4-(phezylmethoxy)phen~l L
L--alycero-pentanamide.
Reaction Scheme A, step (b): To a stirred solution of
O-benzyl-D-valinol (5.42 g, 28.0 mmol) in CH2Clz (15 mL) is
added a 2 M solution of (CH3)3A1/toluene (14.0 mL, 28.0
mmol) over 20 minutes. During the addition more CHZC12 (10
mL) is added to convert the resulting paste to a thin
slurry. As the addition proceeds, a solution again forms
with vigorous gas evolution as it warms to reflux. After
the addition is complete, the solution is allowed to stir
at reflex for IS minutes as gas evolution subsides. The
solution is allowed to cool to room temperature and a
solution of 4-tart-butoxycarbonylamino-2,2-difluoro-3-
hydroxy-5-(4-benzyloxy)phenylpentanoic acid, ethyl ester
(5.38 g, 11.2 mmol, prepared in example 1) in CHSClz (20 mL)
is added over 15 minutes with moderate gas evolution. The
resulting solution is heated at reflex overnight. The
reaction mixture is cooled to 0°C and carefully quenched
with excess methanol. The resulting solution is washed
with 1 N HC1. The aqueous layer is extracted with ether.
The combined organic extracts are washed with water, dried
over anhydrous magnesium sulfate, filtered and concentrated
WO 9610299 ~ I ,' r~ ~ ~ ~ PCTJUS95/07241
_q7_
under vacuum to provide after flash chromatography (4:1
cyclohexane/ethyl acetate) 5.5 g (78~) of title compound
as a beige powder.
Step B
Preparation of [1 (R), 3f~, 4(S)1-4-Amino-2 4 5-trideoxy-
2,2-difluoro-N-[2-methyl-1-[(phenylmethoxy)methyl]propel]-
5-(4-(phenvlmethoxy)phenyl]-L-glvcero-pentanamide.
Reaction Scheme A, step (dj: A solution of the above
prepared alcohol (3.6 g, 5.7 mmolj in HC02H (50 mL) is
allowed to stir at room temperature for 3 hours. The
solution is then concentrated under vacuum at 35°C. The
residue is dissolved in ethyl acetate and aqueous sodium
bicarbonate is added with vigorous stirring. The layers
are separated and the organic layer is washed with water.
The combined aqueous layers are again extracted with ethyl
acetate. The combined organic extracts are dried anhydrous
magnesium sulfate, filtered and concentrated under vacuum
to provide 3.0 g (99~) of the title compound as an orange
oil.
Step C
Preraration of [1 (R), 3E, 4(S)1-2 4,5-Trideoxy-4-([2-
[[(1,1-dimethylethoxy)carbonyl]amino]-3-methyl-1-
oxobutvl]amino]-2,2-difluoro-N-[2-methyl-1-
[(phenylmethoxy)methyl]propel]-S-[4-(phenyl-
methoxv)Phenyl]-L-glycero-pentanamide.
Reaction Scheme D, step (a): The above pregared amine
is coupled under standard conditions in a manner analogous
to that described previously, such as exmaple 2, step A,
with N-(tert-butoxycarbonyl)-L-valine (1.21 g. 5.61 mmol)
to provide, after flash chromatography (9:1 CH2C12/ethyl
acetate), 2.3 g (60~) of title compound as a yellow powder.
~i'O9fV02~t99 ~ ~ ~ C~ ~ L ~ PCTlU595/072~17
-48-
Step D
PtEDaration of [1 fR3 3~F 4(S)]-A-[I2-Amino-3-methyl-i--
_oxobutyl]amino]-2,4 5-trideoxv-2,2-difluoro-N-[2-methyl-1-
j_(~tcenylmethoxyDmethyl ]-prooyl ]-5-[ 4- .
(phenylmet~'foxy)p~'teriyll-L-~11yC2r0-pentanamlde.
Reaction Scheme D, step(b): A solution of the above
prepared amide (2.0 g, 2.8 mmol) in HCOyH (25 mL} is allowed
to stir at room temperature for 6 hours. The solution is
concentrated under vacuum at 30°C and the residue is
IO dissolved in ethyl acetate. Aqueous sodium biearbonate is
added with vigorous stirring. White solids precipitate out
which are filtered and found to be the formats salt of the
(S)-amino alcohol. The organic layer of the filtrate is
separated, washed with water, dried aver anhydrous
i5 magnesium sulfate, filtered and concentrated under vacuum
to provide an amber oil containing some white solids; boor
the (R)- and (S)-amino alcohol and the lactam by-product
are present. The residue is slurred in CHC13 for 1 hour and
the precipitated solids, again the formats salt of the (S)-
2D amino alcohol, are collected. The filtrate is concentrated
under vacuum and the residue dissolved in CHaClz and
additional formats salt of the (S)-amino alcohol is
collected. Flash chromatography of the residual oil (5:1
CHyClz/ethyl acetate to remove the lactam, then 9:1 CHZCIz/
27 CH30H) provides a yellow semi-solid which is the (R)-amino
alcohol contaminated with 5% of the formats of the (S)-
amino alcohol. Yield: 800 mg (43~) of the formats salt of
the (S)-amino alcohol and 640 mg (37~) of the (R)-amino
alcohol. To a stirred suspension of the formats salt of the
a
30 (S)-amino alcohol (800 mg, 1.19 mmol) in ethyl acetate is
added 10~ aqueous sodium bicarbonate. After 2 hours at
roam temperature, the solids dissolve. The organic layer
is separated, washed with water, dried over anhydrous .
magnesium sulfate, filtered and concentrated under vacuum
35 to provide 712 mg (95~) of title compound as a white ,
powder: mp 160-164°C; IR (KBr) umax 3383, 3327. 1676. 1632,
1530, 1514, 1244, 1109 cm-1; 1H NMR (CDC13) d 7.73 (d, 1 H,
-WO 46lQ2q99 ~ ~ ~ ~ ~ f~ ~ PCT/tiS95J07241
-49-
J = 6.9 Hz), 7.43-7.29 (m, 10 H), 7.10 (d, 2 H, J = 8.4
Hz), 6.90 (d, 2 H, J = 8.7 Hz), 6.78 (d, 1 H, J = 9.0 Hz},
5.56 (bs, 1 H), 5.04 (s, 2 H), 4.55 (d, 1 H, J = 12.0 Hz),
4.47 (d, 1 H, J = 12.0 Hz), 4.33-4.18 (m, 2 H), 3.92-3.83
(m, 1 H), 3.60 (dd, 1 H, J = 9.6, 3.6 Hz), 3.45 (dd, 1 Hr
J = 9.6, 3.6 Hz), 3.16 (d, 1 Hr J = 3.9 Hz), 3.03-3.00 (m,
2 H), 2.18-1.93 (m, 2 H), 1.44 (bs, 2 H), 0.94 (d, 6 H, J
= 6.9 Hz), 0.85 (d, 3 H, J = 6.9 Hz), 0.53 (d, 3 H, J =
6.9 Hz); 19F NMR (CDC13) d -110.67 (dd, J = 259, 5 Hz), -
121.74 (dd, J = 258, 18 Hz); mass spectrum, m/z 654 (M+ +
29), 626 (M+ + 1), 325, 92, 91 (100), 72. Anal. Calcd for
C35Hq5F2N305: C, 67.18; H, 7.25; N, 6.72. Found: C, 67.34;
H, 7.21; N, 6.66.
Step E
Preparation of [1 (R), 3~r 4iS)]-2,4,5-Trideoxy-2,2-
difluoro-4-[(3-methyl-2-([3-(4-morpholinyl)-1 3-
dioxopropyl]amino]-1-oxobutyl]amino]-N- 2-methyl-1-
[(phenylmethoxy)methyl]propyl]-5-[4-(phenyl-
methoxy)phenyl)-L-alycero-pentanamide.
Reaction Scheme D, step (c): The (R)-amino alcohol
prepared above (169 mgr 0.270 mmol) is coupled under
standard coupling conditions described previously, such as
example 2, step A, with 2-(4-morpholinylcarbonyl)ethanoic
acid (49 mg, 0.28 mmol, compound (B) prepared in example
5a) to provide, after flash chromatography (3~ CH30H/CHC13),
185 mg (88$) of title comgound as a tan powder: mp 87-94°C;
IR (KB,) vmax 3428, 3325, 1651, 1535, 1512, 1454, 1238,
1115 cm 1; 19F NMR (CDC13) d -116.11 (d, J = 259 Hz), -
119.49 (dd, J = 259, 15 Hz); mass spectrum, m/z 809 (M+ +
29), 781 (M+ + 1) 509, 255, 243, 227, 88 (100).
Step F
Preparation of final title compound.
Reaction Scheme D, step (d): The above prepared
alcohol (177 mg, 0.283 mmol) is oxidized under Swern
conditions in a manner analogous to that described in
w°a ~~ro:~v9 2 ~ c~ c) ~ ~ ~ PcTros9vo~zar
-50-
example 1, step G to provide 170 mg (1D0~)af the final
title compound after flash chromatography (3~ CH3QH/CHCI3).
Recrystallization from CH2C12/ether provides 49 mg (28$) of
final title compound as pale tan powder: mp 119-128°C; IR
(KBr) v,~$X 3306, 1661, 1638, 1539, 1514, 1954, 1238, 1117
cm'1; 1H NMR {CDCI;) d 7.77 and 7.75 and 7.61 and 7.47 (4d,
1 H total, J = 7.1, 7.1, 8.7, 8.9 Hz respectively), 7.43-
7.29 {m, 1D H), 7.16-7.03 (m. 3Hj, 6.91-6.72 and 6.38-6.35
(2m, 3 H total), 5.74 and 5.53-5.36 and 5.24-5.17 (bs and
2m, 1 H total) 5.D2 (s, 2 H), 4.56 and 4.48 (2d, 1 H total,
J = 12.1 Hz), 4.55 and 4.47 (2d, 1 H total, J = 12.0 Hz),
4.27 and 4.23 and 4.12 (add, 1 H total, J = 8.8, 5.3 and
8.7, 5.7 and 7.3, 5.1 Hz respectively), 3.89-3.80 (m, 1 H),
3.69-3.59 (m, 7 H). 3.52-3.90 {m, 3 Hj, 3.32-3.21 (m. 3
H), 2.94-2.75 (m, 1 H), 2.29-1.87 (m, 2 H), 0.94 and 0.93
and 0.92 and 0.91 and D.89 and 0.88 and 0.84 and 0.81 and
0.80 and 0.76 and 0.59 and 0.55 (12d, 12 H total, J = 6.8
and 7.0 and 7.D and 6.9 and 6.9 and 7.0 and 7.0 and 7.0 and
6.8 and 6.8 and 6.9 and 6.$ Hz respectively); i9F NMR
(CDC1;) d -114.46 (d, J = 274 Hz), -112.74 {sj, -113.47 (or
J = 274 Hz), Hydrate: -114.43 (d, J = 253 Hz), -116.47 (d,
J = 254 Hz), -120.63 (d, J = 253 Hz), -122.73 {d, J = 253
Hz); mass spectrum, mjz 807(M+ + 29). 779 (M'~ + 1), 401,
361, 243 (100), 227; exact mass calcd for CqzH53FaNqOg
779.3831, found 779.3878.
35
W09ClU2a99 ~ ~ ~ ~ ~ ~ ~ PCT/US95J07241
-51-
Example 5a
Preparation of 4-Morpholine acetic acid trifluoroacetic
acid salt (A1 and 2-(4-Morpholinylcarbonyl)ethanoic acid
(B).
O O
-CH2 OHsCF3C02H ~ -~ ~~~-OH
CH2
0 B
l0
Preparation of 4-Morpholine acetic acid, 1 1-dimethylethyl
ester.
To a stirred solution of t-butyl bromoacetate (1.61 mL,
10.0 mmol) in tetrahydrofuran (25 mL) is added morpholine
(1.74 mL, 20.0 mmol). The suspension is stirred for 1.5
hours and then concentrated under vacuum. The residue is
dissolved in methylene chloride (50 mL) with saturated
sodium carbonate (50 mL). The layers are separated and the
aqueous is extracted methylene chloride (2 x 25 mL). The
organic extracts are combined, washed with saturated sodium
carbonate (20 mL), brine (30 mL), dried over anhydrous
magnesium sulfate, filtered and concentrated under vacuum.
The residue is triturated with ethyl acetate (15 mL),
filtered and the filtrate concentrated under vacuum to
provide the title compound (2.01 g) as a colorless oil; TLC
Rf = 0.45 (ethyl acetate, silica gel).
Preparation of final title compound (A)
Trifluoroacetic acid (15 mL) is added to the above
prepared ester (1.00 g, 4.97 mmol). The solution is
stirred for S hours and then concentrated under vacuum to
provide a yellow oil. This is triturated with diethyl
ether (25 mLj to provide the final title compound (A) (1.06
82~) as an off white solid; mp 118-121°C.
WO 96i(12~199 PCTiU895I072.ti
~'i~ rl~::i
-sz-
Preparation of 2-(4-Morpho~invlcarbonvl)ethanoic acid,
methyi ester.
To a solution of methylmalonyl chloride (10.0 g, 73.2
mmo1) in methylene chloride (200 mL) at 0°C is added rapidly
dropwise a solution of morpheline (16.0 g, 0.183 mmol, 16.0
mL) in methylene chloride (50 mL). The reaction is stirred
for 4 hours at room temperature. The reaction is then
filtered anti the filtrate is diluted with additional
methylene chloride (200 mL). This is then washed with 1 N
HC1, saturated sodium bicarbonate and brine. The organic
is then concentrated under vacuum to provide a yellow ail,
which is purified by flash chromatography (ethyl acetate,
silica gel) to provide the title compound (9.7 g, 71~) as a
pale yellow oil; Af = 0.28 (ethyl acetate).
Preparation of final title compound (B).
To a solution of the above prepared amide (1.70 g, 9.08
mmol) in methanol (45 mL) is added 1N lithium hydroxide (10
mL, 9.99 mmol). The reaction is stirred at room
temperature for 2.5 hours. The pH is adjusted to 3 with 1N
HCL and the reaction is concentrated under vacuum. The
residue is recrystallized from acetonitrile to provide the
titie'compound (B) (0.216 g, 14$) as a white solid.
30
W9 9fif02499 ~ '~ ~ ~ '~ L ~ PC"fIUS95/d724I
-53-
Example 6
j1R-(1R*,2S*)]-a, a-Difluoro-Y-[[2-(R)-
([(hvdroxy)phenylacetyl]-amino]-3-methyl-1-oxobutyl]amino]-
N-[2-methyl-1-[(phenvlmethoxy)methyl]propel]-6-oxo-4-
(~henylmethoxy)-benzenepentanamide.
Bn
OH
/ ~ ~OBn
w O i w i \
Step A
Preparation of [1 (A), 3~, 4(S)1-2,4,5-Trideoxy-4-[[2-([4-
([2-[([[(1,1-dimethyl-
ethyl)dimethvlsilyl]oxy]phenvlacetvl]amino]-3-methyl-1-
oxobutyl]amino]-2,2-difluoro-N-[2-methyl-1-
((phenylmethoxy)-methyl]propel]-5-[4-fPhenylmethoxy)-
phenyl]-L-qlycero-pentanamide.
Reaction Scheme A', step (e): (1 (R), 3~, 4(S)]-4-
Amino-2,4,5-trideoxy-2,2-difluoco-N-[2-methyl-1-
[(phenylmethoxy)methyl]propyl]-5-[4-(phenyl-
methoxy)phenyl]-L-glycero-pentonamide (0.87 g, 1.6 mmol)
prepared in example 5, step B, is coupled under standard
coupling conditions in a manner analogous to that described
previously, such as example 2, step A, with (R)-O-tert-
butyldimethyl-silylmandelic acid (0.44 g, 1.2 mmol)
prepared in example 1 to provide the title compound, after
flash chromatography (3:2, then 1:l hexane/ethyl acetate),
0.36 g (35~) as a yellow oil: IR (film) Vmax 3405, 3325,
2959, 2932, 2895, 2876, 2861, 1684, 1657, 1512, 1470, 1454,
1244, 1221, 1179, 1098, 864 cm-1: 1gF NMR (CDC13) b -117.20
(d, J = 258 Hz), -118.99 (s), -120.55 (d, J = 262 Hz); mass
spectrum (CI, 70 eV), m/z 874 (M+ + 1), 509, 221 (100):
exact mass calcd for CqgH66FZNgO~Si 874.4638, found 874.4651.
~19~~~~
VVQ 96tt1tq99 PCTfU6951072q 1
-54-
Step B
Preparation of final title compound.
Reaction Scheme A', steps (f) and (g): The above
prepared alcohol{345 mg, 0.39 mmol) is oxidized under Swern
conditions in a manner analogous to that described
previously in example 1 to provide after flash
chromatography (3:2 hexane/ethyl acetate), 154 mg ( 45~) of
the ketone as a yellow glass: IR (film) vex 3408, 3298.
2961, 2932, 2861, 1655, 1613, 1514, 1470, 1454, 1246. 1179,
1098, 1072, 839 cm-l; 1gF NMR (CDG13) c8 -112.6$ (5), -117~29
(d, J = ~53Hz), -120.03 (d, J = 253 Hz); mass spectrum
(CI, 70 eV), m/z 872 (M+ + 1), 221 (IQOj, 197; exact mass
calcd for Cq9H6qFZN307Si 872.4482, found 872.4500.
To a stirred solution of ketone in CHZCla (4 mL) is
added TFA/HZO (9:1, 1 mL). The reaction mixture is allowed
to stir at room temperature for 7 hours and then
ccncentrated under vacuum. The residue is dissolved in
CH2C12 and the solution is washed with saturated aqueous
sodium bicarbonate. brine, dried over anhydrous magnesium
sulfate, filtered and concentrate under vacuum. The
residue is purified by flash chromatography (1:1
hexane,~ethyl acetate) to provide 85 mg (64~) of the final
title compound as a white powder: IR {filmy vmg% 3397, 3314,
2965, 2934, 2874, 1750, 1688, 1663. 1532, 1514, 146$s 1454,
1242, 1179, 1113, 826 cm-1; 19F NMR (CDC13) 8 -112.30 (d, J
= 271 Hz), -113.50 {d, J = 271 H2), -117.54 (dr J = 254
Hz), -119.32 (d, J = 254 Hz); mass spectrum (CI, 70 eV),
m/z 788 (M+ + 1), 107 (I00), 91. Anal. Calcd for
Cq~Hq9FzN30'~0.6H20: C, 67.19; H, 6.58; N, 5.47. Found: C,
67.11; H, 6.55; N, 5.39.
,
W09Gl02J99 ~ ~ ~ PCTJU595/(17241
-55-
Example 7
Preparation of N-[1-([[3.3-Difluoro-4-[[2-methyl-1-
((phenylmethoxy)-methyl]-propyl]aminol-2 4-dioxo-1-[[4-
(phenylmethoxy)phenyl]- methyl]-butyllamino]carbonyl]-2-
methylpropyl]-6-(4-morpholinyl-carbonyl)-3-
pyridinecarboxamide.
O ~ nR.,
O v
JH~ CF H
~H ~ OBn
O O
Step A
Preparation of 6-(4-Morpholinylcarbonyl)-3-
pyridinecarboxylic Acid.
To a stirred 1 M solution of 0-tert-butyl-N,N '-
dicyclohexylisourea [see Mathias, L., Synthesis, 570, 1979,
far preparation] in CH2C12 (20 mL) at 5°C under nitrogen is
added 1.49 g (8.24 mmol) of 2-methyl isocinchomeronate (see
Isagawa, K., et al., Nippon Kagaku Zasshi, 88, 553, 1967,
for preparation] in portions over 2 minutes' After 15
minutes, the cold water bath is removed. As
dicyclohexylurea begins to precipitate, the reaction
mixture is again be placed in a cool water bath at
15-20°C. The mixture is diluted with CH2C12 (5 mL) and
allowed to stir at room temperature overnight. The mixture
is then diluted with CHZC12 (30 mL), filtered, and the blue-
green solids are washed with CH2ClZ . The filtrate and
washings are diluted with ether and washed with dilute
aqueous sodium bicarbonate, water, brine, dried over
anhydrous magnesium sulfate, filtered and concentrated
under vacuum to provide 1.36 g of light blue solids. Flash
chromatography (55:45 cyclohexane/ethyl acetate) provides
1.01 g (52~) of the diester below
W09Gf02494 r ~,1 y PCTIi1S95J07242
-56-
O
l~
CH30 ~J
O
as white crystals. Recrystallization from ether/pentane
provides the above diester as matted white needles: mp 111-
112°C; TR (KBr) V~ax 3420, 2984, 1711, 1379, 1310, 1290,
1246, 1134, 1126, 746 cm~l; TH NMR (CDC13) $ 9.26 (d, 1 H, J
= 2~1 Hz), $.39 (dd, 1 H, J = 8.1, 2.1 Hz), 8.19 (dd, 1 Hr
J = 8.1, 0.7 Hz), 4.04 (s, 3 H), 1.63 (s, 9H); mass
spectrum (EI), m/z 238 (M+ + 1), 237 (M+), 182, 179, 164,
57, (100); CI m/z 238 (M+ + 1), 210, 182 (100). Anal. Calcd
for C12H15N04: C, 60.75; H, 6,37; N. 5.90. Found: C, 60.80;
H, 6.3I; N, 5.75. A solution of the above diester (935 mg,
3.94 mmol) and morpholine (2.0 mL, 23 mmol) in THF (6 mL)
are stirred at reflux under nitrogen for 24 hours.
Additional morpholine (2.0 mL, 23 mmol) is added and
heating continued. After 3 more days, the solution is
concentrated under vacuum and the residue is dissolved in
ethyl acetate and the solution is washed twice with water.
Th,e organic layer is concentrated under vacuum to provide
1.01 g of light yellow solid. Two recrystallizations from
etherJpentane, with filtration through filter aid, provide
542 mg (47%) of amide ester below
O
O~ I \ O
~N
O
as fine cream-colored crystals: mp 91-93°C; IR (KBr) V,~nx
2984, 2965, 1707, 1634, 1370, 1317, 1287, 1169, 1132, 1117
Cm-1; 1H NMR (CDC13) $ 9.13 (dd, 1 A, J = 2.1, 1.9 Hz),
8.36 (dd, 1 H, J = 8.1, Z.l Hz), 7.75 (dd, 1 H, J = 8.1,
0.9 Hz), 3.83 (s, 4 H), 3.73-3.55 (m, 4 H), 1.62 (s, 9 H);
mass spectrum, mfr 293 (M+ + 1), 292, 123, 86 (100). Anal.
Calcd for ClSHZoN2Oq: C, 61.63; H, 6.90; N, 9.58. Found: C,
.wo 9sroza~9 ~ ~ (~ ~ ~ ~ r~ rc~rrus9sro7aai
_57_
61.62; H, 6.91; N, 9.64. HCl gas is bubbled through a
solution of the above amide ester (103.5 mg, 0.354 mmol) in
CH3N0z (4-5 mL) for 20-25 minutes. After standing for an
additional 20 minutes, the solution is concentrated under
vacuum and the residue is triturated with acetone to
provide the title compound (acid) as a pale yellow solid.
The material is combined with crude acid from a similar
experiment (from 502 mg 14) and recrystallized from acetone
to provide (335 mg (69~) of title compound as short, thick
1D white needles: mp 181-183°C; IR (KBr) v~,ax 2928, 2872,
1717, 1601, 1285, 1262, 1111 cm 1; 1H NMR (CD;OD) 8 9.26
(d, 1 H, J = 1.1 Hz), 8.78 (dd, 1 H, J = 8.1, 2.0 Hz), 8.0
(d, 1 H, J = 8.1, Hz), 3.80 (s, 4 H), 3.67 (run, 2 H), 3.51
(nm, 2 H); mass spectrum, m/z 277 (M+ + 41), 265 (M+ + 29),
238, 237 (M+ + 1, 1D0). Anal. Calcd for CylHlzNZOa: C,
55.93; H, 5.13; N, 11.86. Found: C, 56.15; H, 5.32; N,
11.46.
Step B
Preparation of [1 (R1, 3~L 4(S)]-2,4 5-Trideoxy-2,2-
difluoro-4-((3-methyl-2-[[(6-(4-morpholinylcarbonyl)-3-
pyridinyljcarbonyl]aminoj-1-oxobutyl]-aminol-N-(2-methyl-1-
[(phenylmethoxy)methyl)-propylj-5-[4-
_(phenylmethoxy)phenyl)-L-alycero-pentanamide.
Reaction Scheme D, step (c): The (S)-amino alcohol
(185 mg, 0.296 mmol) prepared in example 5, step D, is
cougled with the above prepared acid (77 mg, 0.33 mmol)
under standard coupling conditions in a manner analogous to
that described previously, such as examgle 2, step A, to
provide after flash chromatography (5'k CH30H/CHClg), 227 mg
(91~) of the title compound as a white powder: mp 218-220°C;
IR (KBr) v",ax 3287, 1674, 1663, 1636, 1557, 1539. 1514,
1244, 1115 cm 1; 1H NM.R (CDC1; + DMSO-d6) d 9.05 (d, 1 H, J
= 1.8 Hz), 8.29 (dd. 1 H, J = 8.1, 2.1 Hz), 8.03 (d, 1 H, J
= 9.3 Hz) 7.70 (d, 1 H, J = 8.4 Hz), 7.46 (d, 1 H, J = 8.4
Hz), 7.37-7.22 (m, 11 H), 7.09 (d, 2 H, J = 8.1 Hz), 6.67
(d, 2 H, J = 8.4 Hz), 5.89-5.85 (m, 1 H), 4.88 (d, 1 H, S
wo 9s~oza99 ~ ~ ~ " ~ .~ j rrrms~~ro7aai
J L
-Sa-
= ii.7 Hzj, 4.83(d, 1 H, J = i1.7 Hzjr 4.54 (d, 1 H, J =
i2.0 Hz)r 4.47 (d, 1 H, J = 12.0 Hzj, 4.45-4.35 (mr 1. H),
4.31 (t, 1 H, J = 9.0 Hz), 4.24-4.13 (m, 1 H)r 3.92-3.82
(m, 1 H), 3.77 (s. 4 H), 3.64-3.48 (m, 6 H). 3.08-3.02 (m,
1 H), 2.76 (dd, 1 H, J = 14.4, 10.5 Hz), 2.15-1.92 (m, 2
H), 0.94 (d, 6 H. J = 6.9 Hz), 0.88 (d, 6 H, J = 6.6 Hz):
19F NMR (CDC13 + DMSO-dg) & -109.78 (d, J = 256 Hz), -122.05
(dd, J = 256. 19 Hzj: mass spectrum, mJz 872 (M+ + 29), 844
(M+ + 1), 509 (100), 418, 318. 290. 219. Anal. Calcd for
CQ6HSSFZN50g: C. 65.47: H. 6.57: N, 8.30. Found: C, 65.07:
H, 6.65: Nr 9.12.
Step C
prev~aration_ of final title compound.
Reaction Scheme D, step id3~ mhe above prepared
alcohol (196 mg, 0.232 mmol) is oxidized under under Swern
conditions in a manner analogous to that described
previously in example 1, to provide 196 mg (1000 of crude
final title compound as an orange oil after flash
chromatography (3~ CT330H/CHC13). Recrystallization from
C32C'_2Jpentane, then CH2C12/ ether, provides 100 mg (Sl~s) of
pure final title compound as cream granules: mp 108-135°C:
IR (KBr) vex 3422, 3306, 1638, 1535, 1514, 1.454, 1115 cm-
i; ZH NMR (CBC13) d 8.95-8.87 (m, 1 H), 8.16 and 8.i3 and
8.12-8.08 (2dd and m. I H total, J = 2.2, 1.1 Hz), 7.74 and
7.71 <~d 7.68-7.67 (2dd and m, 1 H total, J = 5.8, 0.7 Hzj,
7.43-7.27 (m, 10 H), 7. I4-7.04 and 7.13 and 7.09 and 7.06
(m and 3d, 2 H total, J = 8.5, 8.8, 8.8 Hz respectively),
6.95-6.77 and 6.88 and 6.85 (m and 2d, 4H total, J = 8.7
Hzj, 6.49 and 6.39 and 6.23 (3d, 1 H total, J = 7.2, 6.7,
9.G Az respectively), 5.60-5.56 and 5.46 and 5.33-5.21 (m
and bs and m, 1 H total), 5.02 and 4.98 and 4.93 (3s, 2 H
total) 4.57-4.40 (m, 3A), 3.81 (bs, 5 H), 3.67-3.59 (m, 5
H), 3.50-3.43 (m, 1H), 3.36-3.27 (m, 1 H)r 2.95-2.76 (mr 1
3~ H), 2.17-1.91 (m, 2 H), 0.96-0.88 and 0.83 and 0.82 and
0.71 and 0.59 (m and 4d, i2 H total, J = 6.8 Hz): 19F NMR
(C~C13) d -111.54 (d, J = 270 Hz), -112.47 (''r J = 269
W0 9(/02x99 PCT/US95/07241
C ~ '~ '~~ ~
-59-
Hz), -113.76 (d, J = 270 Hz), -114.62 (d, J = 270 Hz),
Hydrate: -114.80 (d, J = 253 Hz), -118.53 (s), -118.68
(s), -122.82 (d, J = 259 Hz); mass spectrum, m/z 870 (M+
+ 29), 842 (M* + 1), 197, 194, 91 (100); exact mass calcd
for Cq5H5qF2NgOg 842.3940, found 842.3994. Anal. Calcd for
Casxs3FZNS08: C, 65.62; H, 6.35; N, 8.32. Found: C, 64.80;
H, 6.46; N, 8.27.
Example 8
Preparation of 7.7-Difluoro-4,I2-bis(1-methylethyl)-6,8 11-
trioxo-1-phenyl-9-[[4-(phenylmethoxy)phenyllmethyl]-2-oxa-
5,10,13-triazatetradecan-14-oic Acid. 3-Pvridinvlmethvl
Ester.
Bn
NI ~ O
-' O~NH
~OBn
p ~ O O
Step A
Preparation of N-[(3-Pyridinylmethoxy)carbonylj-L-valine.
To a stirred suspension of L-valine methyl ester
27 hydrochloride (2.0 g, 12 mmol) in toluene (15 mL) is added
triphosgene (1.78 g, 6 mmol) and DMF (0.15 mL, 2 mmol).
The heterogeneous mixture is allowed to stir at room
temperature for 30 minutes. To this mixture is added
tributylamine (0.1 mL, 0.4 mmol) dropwise over 2 minutes.
The reaction mixture is heated to reflux for 1 hour.
Analysis by IR spectroscopy shows a strong signal at 2250
cm-1, indicating the presence of isocyanate. The now
homogeneous solution is cooled to 0°C and 3-pyridylcarbinal
(0.9 mL, 9 mmol) is added dropwise over 10 minutes. A
white precipitate forms during the addition. The reaction
mixture is warmed to room temperature and is allowed to
stir for 16 hours. Additional 3-pyridylcarbinol (1.8 mL,
11d 9fi1U2~9J ~ ~~ ~ ' y ~ PCTII1S95107241
-60-
18 mmal) is added and the cloudy mixture is heated to
reflux for 3 hours. The solution is concentrated under
vacuum, diluted with ethyl acetate and washed with three
portions of water. The solution dried over anhydrous
magnesium sulfate, filtered and concentrated under vacuum
to provide an oil. Flash chromatography (8:1 CH2C12/C2H50H)
of the oil provides 1.46 g {60~) of N-[(3-
pyridinylmethoxy)carbonyl]-L-valine, methyl ester as a
yellow oil: 1H NMR (CDC13) d 8.61 (s, 1 H), 8.56 (d, 1 H, J
= 6.5 Hz), 7.71 (d, 1 H, J = 10.2 Hz), 7.29 (dd, 1 H, J =
8.1, 6.6 Hz), 5.38 {bs, 1 H), 5.13 (s, 2 H), 4.29 {dd, 1 H,
J = 8.2. 5.9 Hz), 2.12 {m, I H), 0.92 (d, 3 H, J = 6.3 Hz),
0.88 (d, 3 H, J = 6.3 Hz); (n]Z~p +9.25° (c 0.85, CHC13).
Anal. Calcd for C13H1aNp04: C, 58.65; H, 6.77; N, 10.53.
Found: C, 57.627 H, 7.03; N, 10.47.
Alternatively the N-[(3-pyridinylmethoxy)carbonyl]-L-
valine, methyl ester can be prepared as follows: L-valine
methyl ester (4.25 g, 34 mmol, freshly prepared ~rom the
hydrochloride salt by neutralization with 50~ aqueous NaOH)
is added to a stirred mixture of 1,1 '-carbonyldiimidazole
(4.86 g, 30 mmol) in CH2Clg {30 mL) over 15 minutes. After
15 more minutes, 3-pyridylcarbinol (5.0 mL, S1 mmol) is
added dropwise to the homogeneous solution. The
resulting solution is heated at 45°C for 3 hours and then
allowed to stir at room temperature overnight. The CH2Cla
is removed under vacuum, and the residue is dissolved in
toluene (70 mL). The solution is heated at 70°C for 6
hours, and then concentrated under vacuum. The residue is
dissolved in CHZC12 and the resulting solution is washed
three times with water, dried over anhydrous magnesium
sulfate, filtered and concentrate to provide an oil, which
is purified by flash chromatograghy to provide 4.6 g (51~)
of the N-[{3-pyzidinylmethoxy)carbonyl]-L-valine, methyl
ester as a yellow oil.
WO 96102499 ~ ~ ~ ~ ~ '~ ~ PCTlUS95l07241
-61-
To a stirred solution of the N-[(3-
pyridinylmethoxy)carbonyl]-L-valine, methyl ester (1.4 g,
5.3 mmol) in CH30H (6 mL) is added LiDH~H20 (0.24 g, 5.8
mmol). The heterogeneous mixture is allowed to stir at
room temperature for 16 hours. The reaction mixture is
diluted with water until completely homogeneous, acidified
with 0.5 N HC1 (11.6 mL, 5.8 mmol) and concentrated under
vacuum to provide a white solid, which is crystallized from
8:1 CHZCIz/CzH50H to the title compound 0.53 g (40%) as a
white crystalline solid: mp 242-244°C; 1H NMR (DMSO-dG) d
8.9 (m, 2 H), B.5 (d, 1 H, J = 8 Hz), 8.05 (m, 1 H), 7.7
(d, 1 H, J = 9 Hz), 5.22 (s, 2 H), 3.9 (m, 1 H), 2.05 (m, 1
H), 0.85 (m, 6 H).
Step B
Preparation of [1 (R), 3~, 4(S)]-2,4,5-Trideoxy-2,2-
difluoro-4-((3-methyl-1-oxo-2-[[(3-
pvridinylmethoxv)carbonyl]amino]butyl]amino]-N-[2-methyl-1-
((phenylmethoxy)methyl]propel]-5-(4-iphenyl-
methoxy)phenyl)-L-alycero-pentanamide.
The amino alcohol prepared in example 5, step D (0.28
g, 0.53 mmol) is coupled with the above prepared acid (0.19
g, 0.74 mmol) under standard coupling conditions described
previously, such as example 2, step A to provide, after
crystallization from ethyl acetate, 0.20 g (50~) of title
compound as a white powder: mp 187-191°C; IR (KBr) vmax
3426, 3300, 2960, 1698. 1674, 1657, 1541, 1512, 1246, 1177,
1151, 1105, 1028, 698 cnil; 1H NMR (DMSO-dg) d 8.58 (d, 1 H,
J = 1.6 Hz), 8.49 (dd, 1 H, J = 4.7, 1.4 Hz), 7.76 (d, 1 H,
J = 7.9 Hz), 7.46-7.24 (m, 13 H), 7.13-7.07 (2d, 2 H total,
J = 8.8 Hz), 6.88 and 6.81 (2d, 2 H total, J = 8.8, 8.3
Hz), 8.34, 7.97, and 6.18 (3d, i H total), 5.08 and 5.D3 (d
and s, 2 H total, J = 5.7 Hz), 4.99 (d, 2 H, J = 2.2 Hz),
4.46 (d, 2 H, J = 3.8 Hz), 4.21 (m, 1 H), 4.06-3.92 (m, 1
H), 3.87-3.73 (m, 2 H), 3.52 (d, 2 H, J = 6.9 Hz), 2.93 (d,
1 H, J = 12.2 Hz), 2.6 (m, 1 H), 1.93-1.75 (m, 2 H), 0.87
(apparent t, 8 H, J = 6.6 Hz), 0.68 (dd, 4 H, J = 6.8, 6.6
~V09fi;112499 1.", 4_ rt . PCTftiS9il07241
~~ I:;A~~
-6 2-
Hz); 19F NMR (DMSO-db) d major diastereomer: -108.9 (dd. J
= 251. 7 Ha), -119.8 (dd. J = 245, 19 Hz), minor
diastereomer: -111.6 (dd, J = 252, 8 Hz), -117.1 (dd. J
= 250, 18 FIz); mass spectrum, m/z 761 (M+ + 1), 555, 138.
121, 110, 92. Anal. Calcd for Cq2HgoF2Na0~: C, 66.32; H,
6.58: N, 7.37. Found: C. b2.43; H, 6.36; N. 6.79.
Step C
Preparation of final title compound.
Reaction Scheme D, step (d): In a manner analogous to
that described previously in example 1, the above prepared
alcohol is oxidized under Swern conditions to provide after
flash chromatography (8:1 CH2Clz/CzH50H), 100 mg (505) of
final title compound as a white powder: mp 108-111°C; IR
(KBr) vmax 3416, 3308, 2963, 1696, 1660, 1537, 1514, 1246,
1115, 1028 cm'i; ;H NMR (DMSO-d6) d 8.91 (d, 1 H, J = 8.8
Hz). 8.58-8.51 (m, 2 H), 7.75 (d, 1 H, J = 7.7 Hz). 7.45-
7.23 (m, i2 H), 7.16 (d. 1 H, J = 8.6 Hz), 6.90(d, Z H, J
= 8.6 Hz), 5.06-4.99 1m. 5 H). 4.45 (d, 2 H. J = 3.1 Hz),
3.92-3.8 (m, 2 H), 3.52 (m, 2 H}, 3.10 (dd, 1 H, J = 14.9.
3.1 Hz), 2.69 (dd, 1 H, J = 14.1, 9.9 Hz}, 1.87 (m, 2 H),
0.84 and 0.78 (2dd, 10 H total. J = 10.8, 7.1 Hz and J =
6.4. 4.8 Hz respectively), 0.60 (dd, 2 H, J = 8.7. 6.9 Hz);
1~C NMR (DMSO-d5) 8 171.2 157.15. 155.8, 149.t7, 148.95,
148.9. 138.3, 137.1, 135.5, 132.6, 130.2, 130.07, 128.8,
128.7, 128.37, 128.1fi, 127.75, 127.67, 127.6, 127.5,
127.44. 127.36. 123.43, 114.5, 71.9, 69.5, 69.45. 69.1,
69.0, 63.16, 59.6, 55.25, 54.76, 54.fi7, 33.8, 33.75, 30.35,
28.8, 28.77. 19.4. 19.33, 18.98, 18.59. 17.86. 29F NMR
(DMSO-dg) d -111.36 (d, J = 272 Hz), -114.36 (d, J = 270
Hz). -112.96 (s). -117.06 (d, J = 255 Hz}, -120.00 (d, J =
250 Hz); mass spectrum. m/z 759 (M+ f 1}, 460, I10, 91.
Anal. Calcd for C42HqgFaNqO~~H20: C, 64.94; H., 6.44; N, 7~22~
Found: C, 64.70; H, 6.36; N, 7.24.
-W096102.t99 ~ ~ r~ ~ ~ ~. ~ PCTIUS95I07241
-63-
Example 9
Preparation of 1R-(1R*,2S*)]-a,a-Difluoro-Y-II3-methyl-2-
[I4-(4-morpholinylsulfonyl)benzoyl]aminol-1-
oxobutyl]aminol-N- 2-methyl-1-
j(phenylmethoxy)methyl]propyll-g-oxo-4-(phenylmethoxy)-
benzenepentanamide.
.,\ OBn
~N
l0 \ ~ \ O
O ~ NH~ CF2 NH
H ~ ~Bn
O ~ O O
Step A
Preparation of [1 (R) 3~, 4(S)]-2,4,5-Trideoxy-2 2-
difluoro-4-[[3-methyl-2-[[4-(4-
marpholinylsulfonyl)benzoyl]amino]-1-oxobutyl]-amino]-N-[2-
methvl-1-[(phenvlmethoxy)methyllpropy11-5- 4-(phenyl-
methoxy)phenyl]-L-glvcero-pentanamide.
Reaction Scheme D, step (c): The (S)-amino alcohol
prepared in example 5, step D (161 mg, 0.26 mmol) is
coupled with the acid (77 mg, 0.28 mmol) of the following
structure
O~ ~ CO2H
~1NI ~O ~ /
S
a
O
(prepared as described by Sycheva, T.P. et al., Shornik
Statei Obshchei Khim, Akad. Nauk S S S R , 1, 568-571
(1953)] under standard coupling conditions in a manner
analogous to that described previously, such as example 2,
step A, to provide after flash chromatography (i:1 ethyl
acetate/CA2C12), 140 mg (62~) of the title compound as a
white solid. Recrystallization from ethyl
acetate/cyclohexane provides the title compound as a white
powder: mp 172.5-190.5°C (172.5-174, then 189.5-190.5°C); IR
~~~i~'C
w~o v6iaiagv r. J rcrws~sra~aa~
-64-
(K.BL) v~,$x 3420, 3322, 2965, 1643, 1534, 1514, 1171. 1113
cm 1; 1H NN1R (CDC13) d 7.90 (d. 2 H. J = 8.4 Hz), 7.79 (d,
2 H, J = 8.3 Hz), 7.4-7.25 (m, 10 H), 7.0B (d, 2 H, J = 8.6
Hz), 6.78 (d overlapping m, 4 H, J = 8.5 Hz), 6.65 (d, 1 H,
J = 8.6 Ha), 4.93 (s, 2 H), 4.53 (d, 1 H, J = 12.0 Hz),
4.47 (d, 1 H, J = 12.0 Hz), 4.45-4.24 (m, 4 H), 3.82 (m, 1
H), 3.73-3.65 (nm, 4 H), 3.62 (dd, 1 H, J = 9.7, 4.1 Hz),
3.46 (dd, 1 H, J = 10.0, 3.9 Hz), 3.1-2.85 (m, 6 H), 2.12
(m, 1 H), 1.98 (m, 1 H), 0.97-0.89 (m, 12 H); i9F NMR
1C (CDC13) d -110.1$ (dd, J = 261, 4 Az), -121.09 (dd, J =
261, 18 Hz); mass spectrum, m/z 879 (M+ + 1, 100), 527,
325. 254. Anal. Calcd for CQ6H56F2N40yS: C, 62.85; H, 6.42;
N, 6.37. Found: C, 62.65: H, 6.43; N, 6.26.
Step B
_Preparation of final title comtround.
Reaction Scheme D, step (d): In a manner analogous to
that described in example I the above prepared alcohol (133
mg, 0.151 mmol) is oxidized under Swern conditions to
2C provide 82 mg of a mixture of diastereomers of final title
ccmpound after flash chromatography {3:2 ethyl
acetate/cyclohexane). The mixture partially crystallizes
from cyclahexane/ethyl acetate upon evaporation of the
solvent. The tacky pale yellow solid is triturated with
ether to provide 59 mg (44~) of a single diastereamer of
final title compound as a white powder: mp 114-116°C; IR
(KHr) vmax 3426, 2971, 1686, 1663, ISI2, 1171, 1115 cm-1;
1H NMR (CDC13) d 7.92 (d, 2 H, J = 8.3 Hz), 7.81 (d, 2 H, J
= 8.3 Hz), 7.41-7.29 (m, i0 H), 7.06 (d, 2 H, J = 8.6 Hz).
6.85 (d, 2 H, J = 8.6 Hz), 6.78 (d, 2 H, J = 8.6 Hz), 6.29
(d, 1 H, J = 6.7 Hz), 5.26 (m, 1 H), 4.99 (s, 2 H, major
peak), 4.55 (d, 1 H, J = 12.0 Hz), 4.48 {d, I H, J = 12.0
Hz), 4.43 (dd, I H, J = 8.4, 6.4 Hz), 3.84 (m, 1 H)r 3.72
(narrow m, 4 H), 3.63 (dd, 1 H, J = 9.8, 3.8 Hz), 3.47 (dd,
1 H, J = 9.7, 3.8 Hz), 3.3.1 (dd, 1 H, J = 14.4, 5.0 Hz),
3.00-2.87 (m, 5 H), 2.14 (m, 1 H), 2.00 (m, 1 H), 0.96 (d,
3 H, J = 6.7 Hz), 0.95 (d, 3 H, J = 6.7 Hz), 0.94 (d, 3 H,
W096I02499 w PCT/US95/072d1
~~~~~~5
-65-
J = 6.7 Hz), 0.92 (d, 3 H, J = 6.7 Hz); 19F NMR (CDC13) d -
112.45 (d, J = 270 Hz), -113.69 (d, J = 270 Hz),-117.94 (d,
J = 255 Hz), -119.29 (d, J = 255 Hz); mass spectrum (CI),
m/z 905 (M+ + 29), 877 (M+ + 1), 525, 507, 401, 353 (100),
197. 107. Anal. Calcd for C46H5dF2NaO9S~0.5 H20: C, 62.36;
H, 6.26; N, 6.32. Found: C, 62.24; H, 6.30; N, 6.27.
Example 10
Preparation of [1R-(1R*,2S*)]-a,a-Difluoro-y-[ 3-methyl-2-
I[4-(4-morpholinylcarbonyl)benzoyl]amino]-1-
oxobutyl]amino]-N-[2-methyl-1-
[(phenvlmethoxv)methyl]propel]-&-oxo-4-(phenylmethoxy)-
benzenepentanamide.
O __
O
JH~ CF NH
~H ~ Bn
~ O O
Step A
Preparation of [1 (R), 3E 4(S)1-2,4 5-Trideoxy-2 2-
difluoro-4-[[3-methyl-2- [4-(4-
morpholinylcarbonyl)benzoyllamino]-1-oxobutyll-amino]-N- 2-
methyl-1-[iphenylmethoxv)methyljpropyl]-5-[4-(phenvl-
methoxy)phenyl]-L-ctlycero-pentanamide
Reaction Scheme D, step (c): The (R) amino alcohol
(171 mg, 0.274 mmol) prepared in example 5, step D is
coupled with 4-(4-morpholinylcarbonyl)benzoic acid (68 mg,
0.29 mmol, prepared in example 10a) under standard coupling
conditions, such as example 2, step A, in a manner
analogous to that described previously to provide, after
flash chromatography (3~ CH30H/CHC13), 217 mg (94~) of the
title compound as a yellow powder: IR (KBr) vm8x 3428,
3322, 1636, 1534, 1512, 1456, 1279, 1258, 1244, 1115 cm 1;
19F NMR (CDC13) d -110.19 (d, J = 259 Hz), -117.26 (d, J =
259 Hz), -120.14 (d, J = 259 Hz), -121.40 (dd, J = 259. 20
~~~°.:)i~~
~'O 9G!(12499 PCT1t1S951072q1
-s6-
Hz) + impurities; mass spectrum, m~'z 871 (M* + 29), 843
(M* + 1), 527, 509, 329, 317 (100), 289, 250, 218.
Step B
Preparation of final title compound.
Reaction Scheme Dr step (d): The above prepared
alcohol is oxidized under Swern conditions as follows: To a
stirred solution of the above prepared alcohol (215 mg,
0.255 mmol) in anhydrous CH2C12 (3 mL) and anhydrous DMSO
l0 (0.36 mL, 5.1 mmol) at -45°C is added 2 M oxalyl
chloride/CH2C12 (1.0 mL) over 5 minutes. The solution is
allowed to stir at -40 to -30°C for 2.5 hours. The solution
is then cooled to -70°C and diisopropylethylamine (0.67 mL,
3.8 mmol) is added over 5 minutes. The solution is allowed
to warm to room temperature, diluted with CHZC12, washed
twice with water, dried over anhydrous magnesium sulfate,
filtered and concentrated under vacuum. The residue is
purified by flash chromatography (3$ CH;OH/CHCI3) to provide
210 mg (98~) of final title compound as an amber oil.
Recrystallization from CHZC12/pentane provides 80 mg (37%)
of final title compound as an amber powder: mp 87-92°C; IR
(K73r) vx,ax 3306, 1636, 1534, 1514, 1454, 1279, 1258, 1244,
1115 cm-i; 1H NMR (CDC13) b 7.82-7.75 (m, 2 H), 7.48-7.30
(m, 12 H), 7.11-7.04 (m, 2 H), 6.91-6.82 (m, Z H), 6.7$ (d,
1 H, J = 8.7 Hz), 6.68(d, 1 H, J = 8.7 Hz), 6.44 and 6.33
(2dr 1 H, J = 6.9 Hz), 5.31-5.22 (m, 1 H), 5.05-4.94 (m, 2
H), 4.57-4.38 (m, 3 H), 3.88-3.28 (m, 1.2 H), 2.94-2.81 (m,
1 H), 2.16-1.93 (m, 2 H), 0.96-0.58 (m, 12 H); 19F NMR
(CDC13) d -lI1.56 (d, J = 270 Hz), -112.56 (d, J = 270 Hz),
-113.63 (d, J = 270 Hz), -114.54 (d, J = 270 Hz), Hydrate:
-113.53 (d, J = 292 Hz), -114.85 (d, J = 252 Hz), -118.71
(d, J = 292 Hz), -122.88 (d, J = 252 Hz); mass spectrum,
m/z 869 (M* + 29), 841 (M* + 1), 525, 317, 279, 218, 197.
157 (100); exact mass calcd for C4~H55F2Nq08 841.3988,
found 841.4043. Anal. Calcd for Cq7HSqHZNqOg: C, 67.13; H,
6.47; N, 6.66. Found: C, 65.80; Hr 6.60; N, 6.69.
W096I(t2-199 ~ ~ ~~ ~ ~ ~ ~ P'CTlUS95/072:11
-67
Example 10a
Prepara'ion of 4-(4-morpholinylcarbonyl)benzoic acid.
N-O '~ ~ O-OH
Preparation of 4-(4-morpholinylcarbonyl)benzoic acid methyl
ester.
To a stirred suspension of monomethyl terephthalate
(25.0 g, 0.14 mmol) in methylene chloride (500 mL) and
dimethylformamide (4 mL) is added dropwise oxalyl chloride
(12.1 mL, 0.14 mol} with vigorous gas evolution. After gas
evolutions ceases, the reaction is stirred for 45 minutes
and then cooled in an ice-water bath. Morpholine (48.4 mL,
0.56 mol) is then added resulting in an exotherm. After
the addition is complete the reaction is allowed to warm to
room temperature and stirred for 1 hour. The reaction
mixture is then washed with 0.5 N HC1 (2 x 500 mL), half-
saturated sodium bicarbonate (2 x 500 mL), water (2 x 500
mL) and brine (300 mL). The reaction is dried over
anhydrous magnesium sulfate, filtered and concentrated
under vacuum to provide the title compound (33.8 g, 98~) as
a white solid; mp 74-76°C.
Preparation of final title compound.
to a stirred solution of the above ester (50.1 g, 0.20
mmol) in methanol (800 mL) is added 1N lithium hydroxide
(241 mL, 0.24 mol) followed by water (160 mL). After
stirring at room temperature for 7 hours, additional water
(100 mL) is added. The reaction is then stirred an
additional 24 hours and then concentrated to remove the
methanol. The aqueous is extracted with methylene chloride
(2 x 100 mL) and diethyl ether (200 mL). The aqueous layer
was then cooled in an ice-water bath and acidified to pH 1
with 12 N HCI (ca. 20 mL). Filtration provides crude white
solid. The solid material is suspended in methylene
chloride (2 L) and heated to reflux. It is then cooled to
W09G1(1i499 ~- r 1 ~7r
. L ~ PCT7U59i107Z-t1
-68-
room temperature, treated with anhgdrous magnesium sulfate,
and filtered. The filtrate is concentrated under vacuum to
provide the final title compound (92.2 g, 89~) as a white
solid; mp 194-196°C.
10
20
30
~WO 9G!02499 ~ ~ ~ ~ ~ ~. ~ PCTlU59510'72.11
-69-
Example 11
Preparation of N-(1-[((3,3-Difluoro-4-[[2-methyl-1-
((phenylmethoxy)methyl]-propyl]amino]-2,4-dioxo-1-[[4-
jphenylmethoxy)phenyl]-methyl)butyl]amino)carbonyl]-2-
methylpropyl)-4-marpholineacetamide.
O
N~NH~ ~OBn
'~~0~( ~v ~ o o ~
O
Step A
Preparation of[1 (R), 3E, 4(S)]-2,4,5-Trideoxy-2 2-
difluora-4-([3-methyl-2-[[(4-morpholinyl)acetyl]amino]-1-
axobutvl)amino]-N-(2-methyl-1-
I(phenylmethoxy)methyl]propyll-5-[4-(phenylmethoxy)phenyl)-
L-qlycero-pentanamide.
Reaction Scheme D, step (c): The (R, S)-amino alcohols
(201 mg, 0.321 mmol) prepared in example 5, step D are
coupled with 4-morpholine acetic acid, trifluoro acid salt
(92 mg, 0.35 mmol, compound (A) prepared in example 5a)
under standard coupling conditions in a manner analogous to
that described previously [with the addition of 1.1 eq. of
NMM] to provide after flash chromatography (4~ CHgOH/CHC13),
224 mg (92~) of title compound as a beige powder: mp 110-
117°C; IR (KBr) vr,gx 3298, 2963, 1674, 1649, 1545, 1512,
1238, 1115, 698 c:a 1; 19F NMR (CDClg) d major diastereomer-
110.56 (dd, J = 260, 5 Hz), -120.79 (dd, J = 260, 18 FIz);
mass spectrum, m/z 781 (M+ + 29), 753 (M* + 1, 100), 752
(M*), 291, 100, 91.
Step B
Preparation of final title compound.
Reaction Scheme D, step (d): In a manner analogous to
that described in example 1 the above prepared alcohol (206
mg, 0.274 mmol) is oxidized under Swern conditions to
W09G1~2499 ~ 1 '~ y.j [ ~ fJ PCTlU59.~~1(172d1
_70_
provide 170 mg (83%) of an orange glass after flash
chromatography (3% CH30H/CHC13). Recrystallization from
CH2Ci2/ether provides 70 mg (34%) of final title compound as
sticky amber crystals: mp 48-53°C; IR (CHC13) vmax 2969,
1686, 1512, 1242, 1117, 760, 750, 731 cm-1; 1H NMR (CDClg)
d 7.55-7.50 (m, 1 H), 7.43-7.29 (m, 10 H), 7.14-7.07 (m, 2
H), 6.92-6.78 (m, 3 H), 6.52-6.49 and 6.40-6.37 (2m, 1 H
total), 5.33-5.19 (m, 1 H), 5.03 (apparent d, J = 3.6 Hz),
4.55 (d, 1 H, J = 11.9 Ha), 4.48 and 4.47 (2d, 1 H total, J
= 1.1.9 Hz), 3.88-3.80 (m, 1 H), 3.74-3.61 (m, 5 H), 3.50-
3.44 (m, 1 H), 3.32-3.25 (m, 1 H), 2.99-2.81 (m, 3 H),
2.49-2.46 (m, 4 H'), 2.14-I.95 (m, 2 H), 1.28-1.24 and 0.95-
0.70 and 0.50 (2m and d, 12 H total, J = 6.7 Hz); 19F NMR
(CDC13) d -111.32 (d, J = 273 Hz), -112.96 (s), -114.64 (d,
J = 272 Hz); mass spectrum. m/z 779 (M+ t 29), 751 (M+ +
1), 226, 100 (100), 86; exact mass calcd for C~;Hg3FZNq07
751.3882, found 751.3892. Anal. Caicd for
CdIH5gF2N407~I.3Hg0: C, 63.60; H, 6.77; N, 7.24. Found: C,
63.64; H, 6.80; N, 6.84.
Example 12
Preparation of N-(1-[([3,3-Difluoro-4-[[2-methyl-1-
j(phenYlmethoxy)methyl]-propS~I]amino]-2 4-dioxo-l~(4-
(phenylmethoxy)phenyl]methyl]-butyl]amino]carbonyl]-2-
methyl~ropY1]-1H-imidazole-1-acetamide.
n
O v
/~~ NH CF NH
iv v
N Tf H ~ Bn
~HCI
O O O
N
Step A
Preparation of 1H-Imidazole-1-acetic Acid.HCl.
To a stirred solution of imidazole (1.50 g, 22.0 mmol)
in CHZClz (25 mLj at 0°C under nitrogen is added drapwise
tent-butyl bromoacetate (1.62 mL, 10.0 mmol). The ice bath
WO 9G/U2.199 ~ ~ ~ .~7 ~ ~ ~ PCT/U595/07241
-71-
is removed and the solution is allowed to stir at room
temperature for 22 hours. The solution is concentrated
under vacuum and the residue is partitioned between ethyl
acetate/ water. The organic layer is washed with water and
concentrated under vacuum to provide 1.22 g (678) of tert-
butyl IH-imidazole-I-acetate as a white crystalline solid:
mg 110-113°C; 1H NMR (CDC13) d 7.49 (s, 1 H), 6.95 (s, 1
H), 4.59 (sr 2 H), 1.47 (s, 9 H). HC1 gas was bubbled
through a solution of the tent-butyl 1H-imidazole-1-acetate
IO in CH3NOi (4-5 mL) for 20-25 minutes. After standing far 20
minutes more, the solution is concentrated under vacuum and
recrystallized from CH3CN/CH30H to provide the title
compound in 70~ yield: mp 206-209°C; 1H NMR (DMSO-dfi) d
9.12(d, 1 H, J = 1.4 Hz), 7.73 (narrow m, 1 H)r 7.68 (d, 1
H, J = 1.4 Hz), 5.15 (s, 2 H). Anal. Calcd for
CSH6N20~~HC1: C, 36.94; H, 4.34; N, 17.23. Found: C,
37.21; H, 4.36; N, 17.39.
Step B
Preparation of[1 (Ri, 3~, 4(S)]-2.4 5-Trideoxy-2,2-
difluoro-4-((2-[(1H-imidazol-1-ylacetyl)amino]-3-methyl-1-
oxobutyl]amino]-N-[2-methyl-1-
((phenylmethoxy)methyl]~ropyl]-5- 4-(phenylmethoxy)phenyl]-
L-glycero-pentanamide.
Reaction Scheme D, step (c): To a solution of the (R)-
amino alcohol (205 mg, 0.328 mmol) prepared in example 5,
step D in anhydrous CHZC12 (4 mL) and anhydrous DMF (7 mL)
is added 1H-imidazole-1-acetic acid~HC1(56 mgr 0.34 mmol)
prepared above and NMM (35 uL, 0.32 mmol). Not all of the
1H-imidazole-1-acetic acid dissolves. HOBT (53 mg, 0.34
mmol) and EDC (66 mg, 0.34 mmol) are then added and the
resulting mixture is allowed to stir overnight at room
temperature. The mixture is concentrated under vacuum and
the residue is dissolved in ethyl acetate. The organic
3~ layer is washed with water, 10~ aqueous sodium bicarbonate,
brine, dried over anhydrous magnesium sulfate, filtered and
concentrated under vacuum to provide yellow solids which
WO961t12q99 ~ .~ t~ ~ ~ l ~ PCTfiIS95107241
-7z-
contain two major spots by TLC (10~ CH30H/CHC13). The two
compounds are separated by chromatography (the first is
eluted with 4~ CH3DH/CHC13, the second with 1:1
CH30H/CFTC13). The less polar material is the formamide of
of the starting material while the more polar material is
the title compound (110 mg, 459) which is isolated as a
white powder: mp I90-193°C: IR (KBr) vmax 3430, 3297, 1678,
1647, 1549. 1512, 1240, 1082 c:a I; 19F NMR (CDC13 + DMSO-d6)
d major diastereomer -115.66 (dd, J = 259, 7 Hz), -120.48
(dd, J = 253. 18 Hz); mass spectrum, m/z 762(M# f 29), 734
(M' + 1), 91 (100).
Step C
Preparation of final title compound.
Reaction Scheme D, step (d): In a manner analogous to
that described in example 1 the above prepared alcohol (101
mg, 0.138 mmol) is oxidized under Swern conditions tc
provide 101 mg (1008) of an orange oil after flash
chromatography (2:1 CHC13/CH30H). Recrystallization from
CH2C12/pentane provides 50 mg (50~) of final title compound
as a beige solid: mp 112-116°C; IR (KBr) vm8x 3428. 3295,
2963, 1647. 1551, 1512, 1240, 1113, 698 cm I; IH NMR (DMSO-
d6) ~ 9.00 and 8.93 and 8.66 and 8.64 (4d, 1 H total, J =
9.0, 9Ø 7.2, 7.2 Hz respectively), 8.16 (apparent
triplet, l H, J = 8.4 Hz), 7.59 (bs, 1 H)r 7.46-7.26 (m, 11
H), 7.15 8nd 7.14 (2d, 1 H total, J = 8.6 Hz), 7.06 (bs, 1
H)r 6.90 and 6.89 (apparent 2d, 3 H, J = 8.7 Hz), 5.05 (s,
2 H), 5.01 (dd. 1 H, J = 7.5. 3.4 Hz), 4.79-4.61 (m, 2 H),
4.50-4.39 (mr 2 H), 4.30-4.20 (m, I H), 3.85-3.76 (m, 1 H),
3.54-3.50 ('m, 2 H), 3.20-3.08 (m, 1 H), 2.72-2.57 (m, 1 H),
2.00-1.72 (m, 2 H), 1.27-1.21 and 0.87-0.69 and 0.61-0.52
(m, 1Z H total); I9F NMR (DMSO-db) 8 -108.87 (d, J = 267
Hz), -109.82 (dr J = 268 Hz), -111.37 (d, J = 268 Hz), -
112.32 (d, J = 254 Hz), 112.58 (d, J = 268 Hz), -117.61 (d,
J = 254 Hz); mass spectrum, mjz 760 (M+ r 29), 732 (M+ +
l), 712, 180, 91 (100), 69; exact mass calcd for CqOHqeF2N506
732.3573, found 732.3541. Anal. Calcd for CqoHq7F2NS06~2H20:
f:
wo ysroa.syy L~ ~ °~ ._j E ~ J rc°rnrs~sro~zal
-73-
C, 62.57; H, 6.69; N, 9.I2. FOUnd: C, 62.07; H, 6.49; N,
9.05.
Example 13
Preparation of [1-[[[3,3-Difluoro-2 4-dioxo-1- 4-
(phenvlmethoxy)phenyl]-methyl]-4-((2-
pyridinylmethyl)aminolbutyl]amino]carbonyl]- 2-
methvlpropyl]-carbamic Acid, 3-Pyridinylmethvl Ester.
N'/ ~ o
~O~ H. X CF2 -NH ~N~
~/ ~ ~/ ~ ~~,~/H
O /' O O
Step A
Preparation of [1 (R) 3~]-2.4 5-Trideoxy-4- [(1 1-
dimethvlethoxy)carbonyl]-amino]-2 2-difluoro-5- 4-
(phenylmethoxy)-phenyll-N-(2-pyridinylmethyl)-L-glycero-
pentanamide.
Reaction Scheme A, step (b): To a stirred solution of
4-tart-butoxycarbonylamino-2,2-difluoro-3-hydroxy-5-(4-
benzyloxy)phenylpentanoic acid, ethyl ester (1.5 g, 3.1
mmol) prepared in example 1 step B, in dry THF (16 mL) is
added 2-(aminomethyl)pyridine (0.38 mL, 3.8 mmol). The
homogeneous solution is heated at reflux for 4 hours and
then allowed to cool to room temperature. A thick white
precipitate forms upon cooling which is filtered and washed
with ether to provide 0.80 g (48~b) of title compound as a
white powder: mp 166-170°C; zH NMR (DMSO-d6) d 9.23 (bs, 1
H), 8.51 (m, 1 H), 7.78 (t, 1 H, J = 7.5 Hz), 7.5-7.25 (m,
8 H), 7.12 (d, 2 H, J = 8.4 Hz), 6.91 (d, 2 H, J = 8.4
Hzj, 6.69 (d, 1 H, J = 10.4 Hz), 6.19 (d, 1 H, J = 8.9
Hz), 5.08 (s, 2 Hj, 4.58-4.32 (m, 2 H), 4.15-3.91 (m, 1 H),
3.82 (m, 1 H), 2.95 (d, 1 H, J = 10.9 Hz), 1.15 (s, 9 H);
19F NMR (DMSO-dbj d major diastereomer: -103.30 (dd, J =
253, 9 Hz), -117.67 (dd, J = 252, 17 Hz), minor
wo vsx~zavv ~ ~ c~ ~ ~ '~ ~ rcTms9~c3~2ai
L
-74-
diastereomer: -111.37 (dd, J = 254, 8 Hz), -121.27 (dd, J
= 2s6, is Hz).
Step H
Preparation of [3~, 4(S}]-2,4,5-Trideoxy-2,2-difluoro-4-
((3-methyl-1-oxo-2-[[(3-
pyridinylmethoxy}carbonyl]amino]butyl]amino]-5-(4-
jphenylmethoxy)phenyl]-N-(2-pyridinylmethyl}-L-qlycero~
pentanamide.
Reaction Schemes A and A', steps {d) and (e): The
above grepared amide (0.2 g, 0.4 mmol) is deprotected in a
mannec analogous to that described previously in example 5,
step H by treatment with HCOyH. The free amine is then
coupled with N-[(3-Pyridinylmethoxy)carbonyl7-L-valine
(0.38 g, 1.5 mmol), prepared in example 8, under standard
coupling conditions in a manner analoqous to that described
previously to provide after recrystallization from ethyl
acetate, 0.09 g (40$) of title compound as a white powder:
mp 201-210°C; 1H NMR (DMSO-dg) d 9.48 (bs, 1 H), 8.79 (m, 1
H}, 8.65 (m, 2 H}, 8.02-7.85 (m, 3 H), 7.39-7.65 (m, 9 H),
7.24 (d, 2 Hr J = 8.5 Hz), 6.96 (d, 2 H, J = 8.5 Hz), 6.48
(d, 1 H, J = 10.4 Hz), 5.25 (d, 2 H, J = 3.8 Hz}, 5.14 {s,
2 H), 4.63-4.55 (m, 2 H}, 4.36 (m, 1 H), 4.17 (m, 1 H),
3.91 (dd, 1 H, J = 7.8, 6.0 Hz) 3.14-3.03 (m, 1 H), 2.81-
2.55 (m, 1 H), 1.95 (m, 1 H), 0.81 (dd, 6 H, J = 9.1, 7.0
Hz); 1vF NMR (DMSO-ds) & -110.17 (dd, J = 254, 11 Hz), -
119.79 (dd, J = 247, 16 Hz).
Step C
Pre oration of final title compound.
Reaction Scheme A', step (f}: The above prepared
alcohol (0.08 g, 0.1 mmol) is oxidized under Swern
conditions in an analogous manner to that described
previously, such as example l, step G, from Z M oxalyl
chlccide/CHZCly (0.6 mL, 1 mmol), DMSO (0.19 mL, 2.6 mmol),
and diisopropylethylamine (0.41 mL, 2.4 mmol), 0.04 g (50~)
to provide the final title compound as a tan powder: mp 90-
wo ~suza99 ~ ~ . ~~ ~ ~ z ~ rcTnrsgsio7za~
-75-
95°C; IR (KBr) v",ax 3401, 3306, 1699, 1599, 1437, 1298.
1242, 1178, 1113, 1026 cm l; 1H NMR (CDC13) d 8.63-8.52 (m,
2 H), 8.42 and 8.19, and 8.05 (3m, 2 H total), 7.77-7.62
(m, 2 H), 7.46-7.22 (m, 9 H), 7.18 (t, 1 H, J = 5.5 Hz),
7.05 (d, 2 H, J = 8.2 Hz), 6.89 (d, 2 H, J = 8.2 Hz), 5.35
and 5.22 (2d, 1 H total, J = 8.5 Hz), 5.10 (s, 2 H), 5.05
(s, 2 H), 4.87-4.66 (m, 1 H), 4.60-4.48 (m, 1 H), 3.81 (dd,
1 H, J = 9.2, 6.7 Hz), 3.32 and 3.27 (2d, 1 H total, J =
5.2 and 4.8 Hz), 3.18 and 3.13 (2d, 1 H total, J = 9.2 and
8.4 Hz) 1.91 (m, 1 H), 1.31 (d, 1 H, J = 6.8 Hz), 0.73 and
0.78 and 0.82 (3d, 5 H total, J = 6.8 Hz); 19F NMR (CDC13) d
-110.48 (d, J = 276 Hz), -113.38 (s), -113.51 (d, J = 276
Hz), Hydrate: -118.75 (d, J = 254 Hz), -120.75 (d, J = 255
Hz); mass spectrum, m/z 674 (M+ + 1), 460, 348, 280, 272,
I5 252, 207, 138, 110 (100)r 92.
Example 14
Preparation of [1-[[(3 3-Difluoro-2,4-dioxo-1-[[4-
jphenylmethoxy)phenyl]-methyll-4- (3-
~'ridinylmethvl)amino]butyl]amino]carbonyl]-2-
methvlpropyl]-carbamic Acid 3-PVridinylmethyl Ester
N~ ~ O a /
N
~O~NH~ CF2 .NH
~/ ~'I( ~/ ~ ~II(H
O ~ O O
Step A
preparation of [1 (R), 302,4,5-Trideoxv-4-f[(1 1-
dimethylethoxy)carbonyll-amino)-2 2-difluoro-5-[4-
phenylmethoxy)-phenyl]-N-(3-pyridinvlmethyl)-L-alycero-
pentanamide.
Reaction Scheme A, step (b): The title compound is
prepared in a manner analogous to that described in example
13, step A from 4-tert-butoxycarbonylamino-2,2-difluoro-3-
~Wb 9(i1~2.199 ~ ~ ~ ~ ~ '~ (') PCTtUS9iJQ7241
L
-76-
hydroxy-5-(4-benzyloxy)phenylpentanoic acid, ethyl ester
(1.5 g, 3.1 mmol) prepared in example i step B, and
3-(aminomethyl)pyridine (0.38 mL, 3.8 mmol). The
precipitated solids are washed with ether and are
recrystallized from ethyl acetate to provide 0.808 {458) of
title compound as a white powder: mp 158-160°C; IH NMR
(CDC13) d 8.55 (bs, 2 H), 7.7 {d, 1 H, J = 7.2 Hz), 7.5-7.2
(mr 8 H), 7.1 (d, Z B, J = 8.4 H2), 6.9 (d, 2 H, J = 8.3
Hz), 5.0 (s, 2 H), 4.9 (d, 1 H, J = 9.8 Az), 4.5 (d, 2 H. J
= 2.6 Bz), 4.1-3.9 (m, 2 H), 3.0-2.8 (m, 2 H), 1.4 (s, 9
H); igF NMR (DMSO-db) d -110.84 {d, J = 272 Ha), -122.56
(dd, J = 273, 21 Hz); mass spectrum, mfz 542 (M~ + 1), 486.
442, 424, 344, 244, 224, 197. 109. 91.
Step B
Preparation of (3~~.4(S)]-2,4,5-Trideoxy-2,2-difluoro-4-[[3-
meth -1-oxo-2-([(3-
pyridinvlmethoxv)carbonyl]aminolbutvl]amino]-5-(4-(phenyl-
methoxy)phenyll-N-(3-pyridinylmethyl)-L-dlycero-
peiltanamide.
Reaction Schemes A and A', steps (d) and (e): The above
prepared amide (0.Z7 g, 0.49 mmol) is treated with 97~
HCOZH. The reaction mixture became homogeneous within 5
minutes. The solution is allowed to stir at roam
temperature foz 3 hours. Zt is then concentrated under
vacuum. The yellow oil is then dissolved in ethyl acetate,
washed twice with aqueous 1 N sodium bicarbonate, dried
over anhydrous magnesium sulfate, filtered and concentrated
under vacuum to provide the corresponding free amine as a
yellow solid (0.18 g, 0.41 mmol). This is dissolved in a
1:1 mixture of CHZC12/ DMF (6 mL). To this stirring
solution at room temperature is added BOBT (0.07 g, 0.5
mmol), NMM {0.05 mL, 0.5 mmol), N-[{3-
pyridinylmethoxy)carbonyl)-L-valine {0.13 g, 0.52 mmol),
prepared in example 8 , and EDC (0.1 g, 0.5 mmol) in that
order. After 16 hours, the mixture is diluted with CHZC12
and washed with water. A white precipitate separates,
WO 96/02499 > ~ ~ ~ ~ ~ ~ PCT/LTS95l07241
_77,-
which is filtered, washed several times with ether, and
recrystallized from ethyl acetate to provide 0.12 g (458)
of title compound as a white powder: mp 195-196°C; 1H NMR
(DMSO-d6) 8 8.7-8.4 (m, 4 H), 7.75 (d, 1 H, J = 10.4 Hz),
7.65 (d, 1 H, J = 11.2 Hz), 7.5-7.3 (m 7 H), 7.15 (d, 2 H,
J = 8.4 Hz), 6.9 (d, 2 H, J = 8.4 Hz), 5.12 (s, 2 H), 5.08
(s, 2 H), 4.5-4.2 (m, 3 H), 4.1-3.9 (m, 2 H), 2.8-2.6 (m, 2
H), 2.0 (m, 1 H), 0.8 (m, 6 H); 19F NMR (DMSO-dg) d -111.2
(d, J = 257 Hz), -122.05 (dd, J = 250, 20 Hz); mass
spectrum, m/z b7b (M+ + 1), 567, 424, 150, 138, 110 (100),
92. Anal. Calcd for Cg6Hg9F2NSOs: C, 63.31; H, 5.87; N,
10.25. Found: C, 63.07; H, 5.90; N, 10.23.
Step C
Preparation of final title compound.
Reaction Scheme A', step (f): The above prepared
alcohol is oxidized under Swern conditions in a manner
analogous to example 1 as follows: To a slightly cloudy
solution of the above prepared alcohol (0.08 g, 0.12 mmol)
in CH2C12 (2 mL) and DMSO (0.21 mL) at -45°C is added 2M
oxalyl chloride/CH2C12 (0.6 mL, 1 mmol) over 5 minutes. The
solution is allowed to stir at -35 to -45°C for 2.5 hours
and is then cooled to -78°C. Diisopropylethylamine (0.42
mL, 2.4 mmol) is added over 5 minutes. The solution is
allowed to warm to room temperature over 1.5 hours, is
diluted with CH2C12 and washed with water. The yellow
precipitate which forms is filtered and washed three times
with ether to provide 0.04 g (50$) of final title compound
as a tan powder: mp 103-105°C; IR (KBr) vmBX 3422, 3295,
2965, 1b97, 1649, 1537, 1512, 1244 cm l; 1H NMR (DMSO-db) d
9.19 (bs, 1 H), 8.61-8.43 (m, 4 H), 7.75-7.59 (m, 2 H),
7.45-7.28 (m, 7 H), 7.09 (d, 2 H, J = 8.5 Hz), 6.81 (d, 2
H, J = 8.5 Hz), 5.09-4.98 (m, 4 H), 4.50-4.26 (m, 3 H),
3.90-3.61 (m, 2 H), 3.12 (m, 2 H), 2.55 (m, 1 H), 1.79 (m,
1 H), 1.27 (t, 5 H, J = 8.6 Hz), 0.77 (t, 1 H, J = 6.2 Hz);
19F NMR (DMSO-dfi) 8 minor diastereomer: -111.34 (apparent
doublet, J = 184 Hz), major diastereomer: -113.24 (d, J =
r
W09fif02499 ~ ~ J ' ~.7 PCTftIS951f172dt
_7$_
254 Hz), -117.56 (d, J = 254 Hz): mass spectrum, mfz 674
(M' + 1}, 565, 460, 252, 207, 143, 138, 110 (100), 92.
Anal. Calcd for C3sH3TF2N5Ds~2HZD~ C, 60~93: H, 5.78: N,
9.87. Found: C, 60.00; H, 6.15; N, 9.56.
Example 15
Preparation of (1-([[3,3-Difluoro-2,4-dioxo-1-I[4-
(phenylmethoxv)-phenyl]-methyl]-4-((2-
pyridinylmethyl)amino]butyl]-amino]carbonyl]- 2-
methvlpropyll-carbamic Acid 2-PVridinylmethyl Ester.
0
O H~ CF2 .NH "wN~
N ~~I(H
O ~ O O
Step A
Preparation of N-[(2-PVridinylmethoxy}carbonyl]-L-valine
Methyl Ester.
In a manner analogous to the preparation of N-[(3-
pyridinylmethoxy)carbonyl[-L-valine in example 8, the title
compound is prepared from L-valine methyl ester
hydrochloride (5.0 g, 30 mmol), triphosgene (4.66 g, 15.7
mmol), and (n-Bu)3N (0.05 mL, 0.4 mmol); however, 2-pyridyl-
carbinol (10.0 mL, 100 mmol, 3.3 equivalents) is added
dropwise to the intermediate isocyanate. Flash
chromatography as in example 8 provides 2.63 g (33g) of
3o title compound as a yellow oil: IR (neat) vmax 3345, 2965,
1726, 1533, 1439, 1314, 1271, 1236. 1213, 11D5 cm-1; rH NMR
(CDC13) d 8.61 (d, 1 H, J = 4.6 Hz), 7.7 (td, 1 H, J = 7.6,
1.8 Hz), 7.38 (d, 1 H, J = 7.7 Hz), 7.27 (dd, 1 H, J = 7.3.
4.6 Hz), 5.45 (d, l H, J = 8.9 Hz), 5.25 (s, 2 H), 4.35
(dd, 1 H, J = 9.2, 4.7 Hz), 3.75 (s, 3 H), 2.24 (m, 1 H),
1.0 (d, 3 H, J = 6.8 Hz), 0.9 d, 3 H. J = 6.8 Hz); 13C NMR
(CDC13) 8 172.4, 156.2, 155.9, 149.4, 136.7, 122.7, 121.6,
SVO 9G/0?~t99 ? ~ t~ ~ ~ ' ~ pC'f/US95I07341
_79-
67.4. 59.1, 52.1, 31.2, 18.9, I7.5; mass spectrum, m/z 267
(M+ + 1, 100), I36, 110.
Anal. Calcd for C13H1gN204: C, 58.65; H, 6.77; N, 10.53.
Found: C, 58.25; H, 6.86; N, 10.46.
Step B
Preparation of N-[(2-PyridinvlmethoxvJcarbonyll-L-valine.
Hydrolysis of the above ester (2.5 g, 9.4 mmol) with
LiOH~H20 (0.79 g, 18.8 mmol) in CH30H in a manner analogous
to that described previously in example 8 provides 2.0 g
(85Ps) of the title compound as a white solid: IR (KBr) vmax
3366, 3065, 2963, 2936, 1722, 1616. 1602, 1577, 1526, 1433,
1279, 1250, 1219, 1109, 764, 627 cm 1; 1H NMR (DMSO-dG) d
8.58 (d, 1 H, J = 4.7 Hzj, 7.96 (t, 1 H, J = 8.0 Hz), 7.6
(d, 1 H, J = 8.0 Hz), 7.43 (d, 1 H, J = 7.5 Hz), 7.4 (dd, 1
H, J = 7.5, 5.2 Hz), 5.15 (s, 2 H), 3.9 (dd, 1 H, J = 8.5,
5.9 Hz}, 2.1 (m, 1 H), 0.92 (dd, 6 H, J = 6.6, 3.7 Hzj; 1~C
NMR (DMSO-dG) d 173.1, 156.3, 156.2, 148.6, 137.4, 123.0,
121.4, 65.9. 59.6, 29.5, I9.1, 18.0; mass spectrum, m/z
253 (M+ + 1), 237, 209, 136 (100), 109, 92, 65.
Step C
Preparation of [3~, 4(S11-2 4,5-Trideoxy-2,2-difluoro-4-
L[3-methyl-1-oxo-2-[[(2-
~ridinylmethoxy)carbonyl]amino]butyllamino]-5-[4-(phenyl-
methoxvlohenyll-N-(2-pyridinylmethyll-L-qlycero-
pentanamide.
Reaction Schemes A and A', steps (d) and (e): The
[1 (R), 3~]-2,4,5-trideoxy-4-[[(1,1-
dimethylethoxy)carbonyl]-amino]-2,2-difluoro-5-[4-
(phenylmethoxy)-phenyl]-N-(2-pyridinylmethyl)-L-glycero-
pentonamide (0.34 g, 0.63 mmol) grepared in example 13,
step A, is deprotected as in example 13, step (d) and
coupled with the above prepared acid (0.21 g, 0.82 mmol)
under standard coupling conditions in a manner analogous to
that described previously to provide 0.14g (30~) of title
compound as a white powder: mp 224-225.5°C; IR (KBr) vmax
W'O 96102a99 PCTIi?595107241
i ~ ~~.'.
_so_
3291, 1697, 1680, 1657. 1618, 1599, 1572. 1537, 1512, 1454.
1439, 1398, 1341, 1300, 1244. 700 cm-l; 1H NMR (DMSO-dg) d
9.31 (bs, 1 H), 7.82-7.71 (m, 3H), 7.48-7.Z3 (m, 10 H),
7.10 (d, 2 H, J = 8.6 Hz), 6.82 (d, 2 A, J = 8.6 Hz), 5.05-
4.95 (m, 4 H), 4.47 (d, 2 H, J = 5.8 Hz), 4.23 (m, 1 Hy,
4.04 (m, 1 H), 3.79 (t, 1 H, J = 6.0 Hz), 4.40 and 3.69
and 3.14 (3m, 1 H total), 2.94 (d, 1 H, J = 13.0 Hz), 2.63
(dd, 1 H, J = 14.6, 10.8 Ha), 1.83 (m, 1 H), 0.70 (dd, 5 H,
J = 9.2, 7.1 Hz), 0.57 and 0.47 (2d, 1 H total, J = 6.9
Hz); 13C NMR (DMSO-d6) d 170.1, 157.4, 156.7. 156.6, 155.7.
148.97, 148.8, 137.2, 136.8, 130.76, 130.21, 128.4, 127.7,
127.5, 122.75, 122.25, 121.0, 120.8, 114.2, 69.0, 66.2,
60.38. 49.9, 44.1, 44Ø 34.0, 30.3, 19.2, 17.95; 19F NMR
(DMSO-ds) d major diastereomer: -110.07 (d, J = 248 Hz), -
119.9 ((d. J = 252 Hz), minor diastereomer: -110.03 (dm, J
= 250 Hz). -118.34 (dm, J = 250 Hz), -111.72 (dm, J = 250
Hz), -123.5 (dm, J = 250 Hzy; mass spectrum. m/z 676 (M*
+ 1), 595, 557, 110, 92 (100y.
Step D
Preparation o~ final title compound.
Reaction Scheme A', step (f): The above alcohol (0.13
g, 0.19 mmol) is oxidized under Swern condition in a manner
analogous to that described previously in example 1 to
provide after flash chromatography (8:1 CHgCl2/G~HgOHy 0.04
g (30%) of a 1:1 mixture of the final title compound as a
tan powder: mp 140-142°C; IR (KBr) vaax 3304, 1695, 1668,
1537, 1514, 1439, 1296. 1244, 1109, 1042 cca l; 1H NMR (DMSO-
d6) d 9.09 and 8.85 and 8.09 (d, m, d, 1 H total), 8.56 (m,
1 H), 8.17 (m, 1 H), 7.78-7.65 (m, 2 H), 7.45-7.15 (m, 10
H), 7.05 (d, 2 H, J = 8.6 Hz), 6.89 (d. 2 H, J = 8.6 Az),
5.56 and 5.43 (2d, I A total, J = 9.2, 9.2 Hz), 5.30-4.95
(m, 5 H), 4.84-4.45 (m, 2 H), 3.85-3.7 (m, 1 H), 3.32-3.27
(2d,1 H, J = 4.9, 4.0 Hz), 3.16-2.80 (2m, 1 H), 1.90-1.70
(m, 1 H), 0:83-0.75 (m, 5 H), 0.56 (d, 1 H, J = 6.8 Hz); 19F
NMR (DMSO-ds) 8 -110.07 (d, J = 275 Hz), -113.25 (agparent
d. J = 9 Hzy, -113.45 (d, J = 276 Hz), Hydrate: -118.5 (d,
~ly~f~Y~S
W'O 9Gln:al9 PCTIUS95lD7241
-81-
J = 257 Hz), -121.00 (d, J = 253 Hz); mass spectrum, m/z
674 (M+ 1), 460, 143, 110, 92.
10
20
30
W09GI02-i99 ~ ~ ~ ~ .~ ~ 5 PCTYUS95f0724!
_82_
Example I6
Preparation of [1-[[[3 3-Difluorc-2 4-dioxo-1-[[4-
~phenvlmethoxyi-phenyl]-methyl]-4-C(3-
pyridinylmethyl)amino]butyl]-amino]carbonyll- 2-
methvlpropvl]-carbamic Acid 2-Pyridinylmethyl Ester.
/ \ o . .~ / \n]
l0 N O H CF2 NH
N
O ~ O O
Step A
Preparation of (3~, 4(S)]-2,4,5-Trideoxy-2,2-di.fluoro-4-
Cf3-methyl-1-oxo-2-([(2-
yridinylmethoxy)carbonyl]amino]butyl]amino]-5-[4-(phenyl-
methoxy)phenyl]-N-(3-pyridinylmethyl)-L-c7lycero-
pentanamide.
Reaction Schemes A and A', steps (d) and (e): The [1
(R), 3L]-2,4,5-trideoxy-4-[[(1,1-dimethylethoxy)carbonyl]-
amino]-2,2-difluoro-5-[4-(phenylmethoxy)-phenyl]-N-(3-
gyri.dinylmethyl)-L-glycero-pentonamide
(0.357 g, 0.709 mmol) prepared in example 14, step A. is
deprotected as in example 14, step (d) and coupled with N-
[(2-pyri.dinylmethoxy)carbonyl]-L-valine (0.157 g, 0.620
mmol) prepared in example 15, under standard coupling
conditions described previously to provide after treating
the crude material with hot CH30H, filtering, and
concentrating under uacuum, 180 mg (54%) of title compound
as a white powder: IR (KBr) v~,ax 3298, 1697, 1680, 1659,
1537, 1512, 1298. 1244, 1103 cm-1: 1R Nt2R (Dt'LSO-ds) 8 9.36-
9.32 (m, 1 H), 8.52-8.46 (m, 3 H), 7.82-7.68 (m, 3 H),
7.41-7.26 (m, 7 H), 7.19 (d, 1 H, J = 9.6 Hz), 7.07 (d, 2
g~ J = 8.4 Hz), 6.81 (d, 2 H, J = 8.4 Hz), 6.Z7 (d, 1 H, J
= 7.2 FIz), 5.12 (d, 1 H, J -- 13.5 Hz), 5.06 (d., 1 H, J =
13.5 Hz), 5.02 (d, 1 H, J = 12.0 Hz), 4.97 (d, 1 H, J =
I l ,.I V L ,J
W 0 9(10299 PCTIUS9510724I
-83-
12.0 Hz), 4.44 (dd, 1 H, J = 15.3, 6.0 Hz), 4.35 (dd, 1 H,
J = 15.3, 5.7 Hz), 4.24-4.15 (m, 1 H), 4.1I (dd, 1 H, J =
10.5, 5.1 Hz), 4.06-3.95 (m, 1 H), 3.79 (dd, 1 H, J = 9.3,
6.9 Hz), 2.93-2.88 (m, 1 H), 2.60 (dd, 1 H, J = 13.8, 11.1
Hz), 1.82-1.77 (m, 1 H), 0.71 (d, 3H, J = 6.9 Hz), 0.68 (d,
3H, J = 6.9 Hz}; 1~F NMR (DMSO-dfi) 8 -109.87 (dd, J = 252, 8
Hz), -120.07 (dd, J = 253, 17 Hz}; mass spectrum, m/z 704
(M+ + 29), 67b (M+ + 1), 424, 241, 138, 110, 92 (100).
Anal. Calcd for C36H3gF2N506: C, 63.99; H, 5.82; N, 10.36.
1D Found: C, 60.62; H, 5.97; N, 9.83.
Step B
Preoaratioa of final title compound.
Reaction Scheme A', step (f): In a manner analogous to
that described previously in example 1, the above prepared
alcohol (152 mg, 0.225 mmo1) is oxidized under Swern
conditions to provide after flash chromatography (19:1
CHClg/CH3OH) 104 mg (688} of final title compound which is
recrystallized twice from CH2C12/pentane to provide 34 mg
(22~) of final title compound as a cream powder: mp 97-
102°C; IR (KBr) vmax 3405, 3324, 1699, 1534, 1512, 1437,
1242, 1178, 1113 cm-1; 1H NMR (CDC13) d 8.56-8.51 (m, 3H),
7.71-7.62 (m, 2 H), 7.43-7.19 (m, 10 H), 7.10-7.05 (m, 2
H), 6.91-6.83 (m, 2 H), 6.48-6.37 (m, 1 H}, 5.48-5.10 (m,
3H), 5.02 and 5.00 (2s, 2 H total), 4.61-4.46 (m, 2 H},
3.90 and 3.87-3.81 (dd and m, 1 H total, J = 8.7, 6.1 Hz),
3.36-3.23 (m, 1 H), 2.91-2.73 (m, 1 H total), 2.07-1.88 (m,
1 H), 0.86 and 0.79 and 0.76 and 0.75 and 0.61 and 0.59
(6d, 6 H total, J = 6.6 Hz); 19F NMR (CDC1;) d -111.65 (d, J
= 277 Hz), -111.73 (d, J = 276 Hz), -113.15 (d, J = 276
Hz), -113.20 (d, J = 277 Hz), Hydrate: -115.64 (d, J = 257
Hz), -116.55 (d, 3 = 256 Hz), -120.42 (d, J = 256 Hz), -
121.55 (d, J = 256 Hz); mass spectrum, m/z 702 (M+ + 29),
674 (M+ + 1), 138, 110, 92 (100). Anal. Calcd for
C36H37F2N506: C, 64.18; H, 5.54; N, 11.87. Found: C, 61.96;
H, 5.80; N, 10.38.
WO 961112.199 ~ ~ ~ ~ ~ ~ ~ PCTIUS95~~172d1
-84-
Example 17
Preparation of N-14-(N-{2-pyridylmethyl}oxycarbonyl-L-
valyl)am~na-2 2-difluora-1 3-dioxo-5-(4-benzyloxy)phenyl-
pentyl]-O-(3-pyridylmethyl)-D-valinol.
O
O H CF2 NH
~H ~ °
O~ o O
Step A
Preparation of N-tritvl-D-valinol.
A solution of D-valinol (4.95 g, 48.06 mmal),
triethylamine (7.4 mL, 52.87 mmol) and trityl chloride
(14.74 g, 52.87 mmol) in dry dichloromethane (75 mL) is
stirred for 17 hours at room temperature. The organic
solution is washed with water (2 x 75 mL), dried aver
anhydrous sodium sulfate, filtered and concentrated under
vacuum. The resulting oil is purified by flash
chromatography (silica gel, ethyl acetate/petroleum ether,
15/85) to provide the title compound (1.3.5 g, 81~;); (1f=0.45
(ethyl acetate petroleum ether. 15/85).
Step B
Preparation of N-trityl-O-3-pyridvlmethvl-D-valinol.
Under a nitrogen atmosphere, to a suspension of sodium
hydride (1.3 g, 30 mmol, 55~ dispersion in oil, previously
washed twice with pentane) in dry DMF (3 mL) is added with
stirring, a solution of N-trityl-D-valinol (3.45 g, i0
mmol) in DNdF (23 mL). The reaction is stirred for 30
minutes at room temperature and then cooled to 0°C and
tetrabutylammonium iodide (0.37 g, 1 mmol) is added. Then
3-picolyl chloride HC1 {1.81 g, 11 mmo1) is added to the
reaction in portions over 5 minutes. After addition is
WO 96f02~99 ~ ~ ~ ,~ ~ ~ J PCTlUS95/07241
_85-
complete, the cooling bath is removed and the mixture
allowed to stirr for I7 hours at room temperature. The
reaction mixture is then cooled with an ice bath and
hydrolyzed with water (100 mL). The mixture is then
extracted with ethyl acetate (2 x 100 mL). The organic
extracts are washed with water (2 x 50 mL), combined, dried
over anhydrous sodium sulfate, filtered and concentrated
under vacuum. The resulting yellow oil is gurified by
flash chromatography (silica gel, dichloromethane/ethyl
acetate, 9/1, Rp=0.42) to provide the title compound (3.4 g,
78B) as an oil.
Step C
Preparation of O-3-pyridvlmethyl-D-valinol.
A solution of N-trityl-O-3-pyridylmethyl-D-valinol
(3.63 g, 8.3 mmol) in formic acid (30 mL) is kept for 5.5
hours at room temperature. The formic acid is removed
under vacuum and the residue is dissolved in water (100
mL). The mixture is extracted with ethyl acetate (100 mL,
50 mL) to remove the trityl alcohol. The aqueous phase is
then made basic with saturated sodium carbonate (50 mL) and
4N sodium hydroxide (3 mL). The aqueous mixture is then
extracted with ethyl acetate (4 x 50 mL). The organic
extracts are rinsed with brine (2 x 50 mL), combined, dried
over sodium sulfater filtered and concentrated under vacuum
to grovide the title compound (1.32 g, 82g) Rg=0.12 (silica
gel, dichloromethane/methanol, 8/2).
Step D
Preparation of N-[4-tert-butoxycarbonylamino-2,2-difluoro-
3-hydroxy-1-oxo-5-(4-benzyloxv)phenyl-pentylj-O-(3-
ridylmethyl)-D-valinol.
Reaction Scheme A, step (b); A solution of the ester
(1.14 g, 2.38 mmol, prepared in examgle 1, step B) and of
O-3-pyridylmethyl-D-valinol (1.32 g, 6.8 mmol, prepared in
step C above) in dry tetrahydrofuran (1.5 mL) is heated for
2 days under reflux. After cooling, the reaction mixture
WO 9G102~99 P~"1°~S9e/07241
2~~j~1~5
-ax-
is diluted with ethyl acetate (5 mL), pentane (10 mL) and
the precipitate is collected by.filtration. The filtrate
is rinsed with pentane and recrystallized from
dichloromethane/methanol/gentane to provide the title
compound (0.8 g, 54~) as a white solid, Rg=0.5 (silica gel,
ethyl acetate); mass spectrum, m/z 628 (t~+).
Step E
Preparation of N-[4-amino-2,2-difluoro-3-hydroxy-1-oxo-5-
~N-(benzyloxy)rshenyl-pentyl]-O-(3-pyridylmethyl)-D-valinol.
Reaction Scheme A, step (d); The title compound is
obtained in 91~ yield following the deprotection procedure
in an analogous manner described in Example 17, step C;
mass spectrums m/z 528 (MH+).
Step F
Preparation of N- 4-(N-(2-pyridvlmethyl~oxycarbonyl-L-
valyl)amino-2 2-difluoro-3-hydroxy-1-oxo-5-(4-
benzyloxv)phenyl-pentyll-O-(3-oyridylmethyl)-D-valinol.
v~ Reaction Scheme A', step (e); To a solution of N-(2-
pyridylmethyloxycarbonyl)-L-valine (0.101 g, 0.4 mmol,
prepared in Example 15, step 8), in anhydrous DMF (2 mL)
are added under an atmosphere of nitrogen, N-
hydroxybenztriaxole hydrate (0.115 g, 0.4 mmolj, 1-ethyl-
3(3-dimethyiaminopropyl)carbodiimide (0.085 g, 0.44 mmol)
and DMF (1 mL). The reaction mixture is allowed to stir
for 30 minutes at room temperature and N-[4-amino-2,2-
difluoro-3-hydroxy-1-oxo-5-(N-(benzyloxy)phenyl-pentyl]-O-
(3-pyridylmethyl)-D-valinol (0.211 g, 0.4 mmol, prepared
above) in DMF (1 mL) is added. The reaction is allowed to
stir for 15 hours, then diluted with ethyl acetate (80 mL)
and washed with water (2 x 80 mLj. The aqueous rinses are
extracted with ethyl acetate ;80 mLj. The combined organic
extracts are dried over anhydrous sodium sulfate, filtered
and concentrated under vacuum. The residue is purified by
flash chromatography (2x, silica gel,
1V0 9610299 ~ ~ ~ w~ j ~l ~ PCTlUS951a7241
_87-
dichloromethane/ethanol, 95/5, RE=0.15) to provide the title
compound (0.170 g, 56%); mass spectrum, m/z 762 (MH+).
Step G
Preparation of final title compound.
Reaction Scheme A', step (f); Ta a solution of oxalyl
chloride (0.195 mL, 2.23 mmol) in anhydrous dichloromethane
(0.5 mL) at -60°C is added slowly under an atmosphere of
nitrogen, freshly distilled dimethyl sulfoxide (0.316 mL,
4.46 mmol) in dichloromethane (1.5 mL). After 10 minutes
of stirring at -60°C, the temperature is allowed to rise to
-15°C. A mixture of a solution of the alcohol prepared
above (0.170 g, 0.223 mmol) in dichloromethane (7 mL) and
dimethylsulfoxide (0.5 mL) is added to the reaction
dropwise. The reaction is then stirred for 1.75 hours at -
15°C and then cooled to -78°C. Diisopropylethylamine (0.93
mL, 6.69 mmol) is added and the reaction is stirred an
additional 10 minutes. The reaction is then allowed to
warm to room temperature and is diluted with
dichloromethane (25 mL). The reaction mixture is washed
with water (2 x 25 mL). The aqueous washes are extracted
with dichloromethane (25 mL). The combined organic
extracts are dried over anhydrous sodium sulfate, filtered
and concentrated under vacuum. The residue is purified by
flash chromatography (silica gel, ethyl acetate, Rf=0.17)
followed by recrystallization from dichloromethane/pentane
to provide the title compound (0.12D g, 71%); mass
spectrum, m/z 76D (MH+); 1H NMR (DMSO-d6) d 9.12-8.96 (m,
1H), 8.66-8.55 (m, 2H), 7.96-7.85 (m, 1H), 7.85-7.75 (m,
1H), 7.60-7.35 (m, 7H), 7.30-7.19 (m, 2H), 7.08-6.83 (m,
2H), 5.20 (s, 2H), 5.17-5.05 (m, 3H), 4.65-4.53 (m, 2H),
4.07-3.87 (2m, 2H), 3.72-3.60 (m, 2H), 3.28-3.14 and 2.86-
2.78 (m, 2H), 2.09-1.83 (m, 2H), 1.27-0.67 (m, 12H); 19F NMR
(DMSO-db, T=60°C) & (CgF6) 52.94 (d, J=269 Hz), 52.30 (d,
J=270 Hz), 51.25 (d, J=270 Hz), 50.35 (d, J=269 Hz).
Anal. Calcd for CqlH~7N507F2, 0.25 HZO: C,.64.43; H, 6.26; N,
9.16;
1V0 9E/(12-199 PC'T/US9SI07241
J ~ ~?~ j I L_
-$8-
Found C, 64.24; H, 6.25; 2d, 9.01.
Example 18
Dreparation N-(4-(N-{3-pyridvlmethyl}oxvcarbonyl-L-
valylDamino-2 2-difluoro-1,3-dioxo-5-(4-benzyloxy)phenyl.-
oentyl)-O-(3-pyridylmethyl)-D-valinol.
N
Step A
Dreparation of N-I4-tN-~3-pyridylmethyl}oxycarbonyl-L-
valyl)amino-2 2-difluoro-3-hvdroxy-1-oxo-5-(4-
benzyloxy)phenyi-pentyl)-0-(3-pyridylmethyl)-D-valinol.
Reaction Scheme A', step (e): The title compound is
prepared in a manner analogous to the coupling procedure
described in Example 17, Step Fr utilizing the amine
prepared in Example 17, Step E, and the acid prepared in
Example 8, Step A. The title compound is obtained in 84~k
yield, Rg=0.43 (silica gel, dichloromethanejethanol, 95j5)~
mass spectrum, m,~z 762 (MH+).
Anal. Calcd for C4lHqgNg0~F2~0.5 H20: C, 63.88; H, 6.54: N.
9.08;
Found C, 63.87: Hr 6.47; N, 9.06.
3G Step B
Preparation of Finai Title Compound.
Reaction Scheme A', step (f); The title compound is
prepared in a manner analogous to the Swern Oxidati.an
described in Example 17, Reaction Scheme A', step (f),
utilizing the above alcohol. The title compound is
obtained in 50~ yield, Rf=G.1 (silica gel, ethyl acetate):
1H NMR (DMSO-d5) 8 9.04-8.90 (m, 1H), 8.71-8.50 (m, 3H),
7.90-7.70 (m, 2H), 7.60-6.85 (m, 12H), 5.27-5.02 (m, 5H),
r y
WO 96!02.199 j
.~ ; L J PCT1U595107241
_89_
4.50-4.66 (m, 2H), 3.85-4.07 (m, 2H}, 3.72-3.57 (m, 2H),
3.30-3.12 and 2.85-2.65 (m, 2H); 2.05-1.8 (m, 2H), 1.20-
0.59 (m, 12H); 19F NMR (DMSO-dg) d (CgFg) 52.93 (d, J=268
Hz), 52.15 (d, J=269 Hz), 50.92 (d, J=269 Hz), 49.68 (d,
J=268 Hz); hydrates, 49.31 (d, J=254 Hz), 48.31 (d, J=254
Hz), 46.22 (d, J=254 Hz), 44.68 (d, J=254 Hz).
Anal. Calcd for Cq1H47N507F2, H20: C, 63.31; H, 6.35; N,
9.00;
Found C, 62.56; H, 6.26; N, 8.66.
Example 19
Preparation of N-(4-(N-{3-pyridylmethyl}oxyearbanvl-L-
valyl)amino-2,2-dilluoro-1,3-dioxo-5-(4-benzyloxy)phenyl-
pentyll-O-methyl -D-valinol.
S \ a
N
O H~ CF2 NH
H ~ CH3
O ~ O O
Step A
Preparation of N-tert-butoxycarbonyl-D-valinol.
A solution of D-valinol (5.1 g, 49.4 mmol) and di-tert-
butyldicarbonate (10.9 g, 50 mmol), in methanol (60 mL) is
stirred for 17 hours at room temperature. The reaction is
concentrated under vacuum and the residue is purified by
flash chromatography (silica gel, ethyl acetateJpetroleum
ether, 3/7, R~=0.37) to provide the title compound in
quantitative yield (10.07 g) as a colorless oil; mass
spectrum, m/z 204 (MH+).
1V0 961i12.i99 PCTfUS951072~tt
-90-
Step B
Preparation of N-tert-Butoxycarbonyl-O-methyl-D-valinol.
To a solution of N-tert-butoxycarbonyl-D-valinol (5.075
g, 25 mmol) and methyl iodide (27.5 mL of a 1M solution in
tetrahydrofuran, 27.5 mmol) in dry tetrahydrofuran (45 mL)
under an atmosphere of nitrogen at 0°C, is added with
stirring, potassium-tert-butoxide (3.086 g, 27.5 mmol) in 3
portions. The reaction mixture is kept at room temperature
for 5.5 hours, then hydrolyzed with a saturated solution of
sodium chloride and extracted with ethyl acetate (2 x 300
mL). The organic extracts are washed with brine (2 x 300
mL), combined, dried over anhydrous sodium sulfate,
filtered and concentrated under vacuum. The residue is
purified by flash chromatography (silica gel, ethyl
acetate/petroleum ether, 1/9, Rf=0.25) to provide the title
compound (2.93 g, 54%) as an oil.
Step C
Preparation of O-methyl-D-valinol.
A solution of N-tert-butoxycarbonyl-O-methyl-D-valinol
12~93 g. 13.5 mmol) in dry ether saturated with hydrogen
chloride (11 mL) is stirred at room temperature for 3.5
hours. The reaction is then concentrated under vacuum.
Pentane (50 mL) is added to the residue, followed by
addition of diethylamine (7 mL). The mixture is then
stirred for 10 minutes and the solid is removed by
filtration. The filtrate is concentrated under vacuum(150
bars) at room temperature to provide the title compound
(1.30 g, 82%) as a colorless oil.
Step D
Preparation of N-[4-tert-butoxvcarbonvlamina-2.2-difluoro-
3-hydroxy-1-axo-5-(4-benzyloxy)Phenyl-pentyl]-O-methyl-D-
valinal.
Reaction Scheme A, step (b); The title compound is
prepared in a manner analogous to the procedure of Example
i7. Step D, from the ester of Example 1, step B and the
WO96102.199 ~ ~ ~ '~) ~ ~ ~ PCT/US95/07241
-91-
above pzepared O-methyl-D-valinol followed by flash
chromatography (78$ yield); Rf=0.19 (minor) and 0.09
(major) (silica gel, ethyl acetate/petroleum ether, 35/65);
mass spectrum, m/z 551 (MH+), 568 (MNHq+).
Step E
Preparation of N-[4-amino-2,2-difluoro-3-hvdroxy-1-oxo-5-
(4-benzyloxy)phenyl-pentyi]-O-methyl-D-valinol.
Reaction Scheme A, step (d); The title compound is
prepared in a manner analogous to the deprotection
procedure of Example 17, step C, from N-[4-tert-
butoxycarbonylamino-2,2-difluoro-3-hydroxy-1-oxo-5-(4-
butyloxy)phenyl-pentyl]-O-methyl-D-valinol
prepared above (quantitative yield); mass sgectrum, mJz 451
(MH+).
Step F
Preparation of N-[4-(N-{3-pyridvlmethvl}oxycarbonvl-L-
valyl)amino-2,2-difluoro-3-hydroxy-1-oxo-5-(4-
benzyioxy)phenyl-pentyl]-O-methyl-D-valinol.
Reaction Scheme A', step (e); The title compound is
prepared in a manner analogous to the coupling method
described in Example 17, step F, from N-[4-amino-2,2-
difluoro-3-hydroxy-1-oxo-5-(4-benzylaxy)phenyl-pentyl)-O-
methyl-D-valinol prepared above and the acid prepared in
Example 8, Step A. Purification by crystallization (ethyl
acetate plus 108 ethanol/gentane) provides the title
compound in 58~ yield; mass spectrum, m/z 685 (MH+).
Steg G
Preparation of final title compound.
Reaction Scheme A', step (f); The final title compound
is prepared in a manner analogous to the Swern Oxidation
described in Example 17, step G from N-[4-(N-{3-
pyridylmethyl}oxycarbonyl-L-valyl)amino-2,2-difluoro-3-
hydroxy-1-oxo-5-(4-benzyloxy)ghenyl-pentyl]-O-methyl-D-
valinol prepared above (15~ yield, Rf=0.1, silica gel,
wo mraa.~vv L ~ r~, ~~ ~ , rrrrL~svseo~aa~
,.:,~J
-92-
dichloromethane/ethyl acetate, 4/6): 1H NMR (DMSO-d~) 5
9. a7-8.93 (m, 1H), 8.87-8.50 (m, 2H), 8.40-8.31 (m, 1H),
7.94-7.81 (m, 1H), 7.60-6.59 (several m, I1H), 5.29-5.01
(m, 5H), 4.10-3.78 (m, 2H',), 3.58-3.43 (m, 2H), 3.35-3.25
(several s, 3H), 3.30-3.08 and 2.85-2.63 (m, 2H), 2.11-
1.72 (m, 2H), 1.04-D.48 (m, 12H); 19F N~iR {DMSO-dg) d (CsF~)
53.27 (d, J=267 Hzj, 52.60 (d, J=269 H2), 51.32 {d, J=269
Hz), 49.76 (d, J=267 Hz), 49.89 (d, J=254 Hz}, 48.61 (d,
J=254 Hz}, 46.49 {d, J=254 Hz), 44,73 {J=2S4 Hz); mass
spectrum, mfz 683 (MH+).
Anal. Calcd for C36HqqNqO~F2, HyO: C, 61.70; H, 6.62; N,
7.99:
FOUnd Cv 62. D7: H, 6.46: N, 7.84.
IS Example 20
Preparation of N-{4-(N-~3-pyridylmethyl}oxycarbonyl-L-
valyl)amino-2 2-difluoro-1,3-dioxo-5-(4-benzylaxv}phenvl-
pentyll-O-(2-pyridylmethyl)-D-valinol.
za j
o ',
N
O' f!H~-~ tF2~NH
'..' N H O
O ~ O O N
2~ i
Step A
Preparation of N-tritvl-O-2-pyridylmethyi-D-valinol.
The title compound is prepared in a manner analogous
to the procedure described in Example 17. step B from the
alcohol of Example 17, step A and 2-picolyl chloride HC1,
in 81% yield, Rp=0.52 (silica gel, dichloromethane/ethyl
acetate, 9f1).
Step B
Preparation of O-2-pyridylmethyl-D-valinol.
The title compound is prepared in a manner analogous to
the procedure described in Example 17. step C from N-
R'O 9~f02-199 ~ ~ ~ ~ ~ 'j ~ PCT/IiS95107241
_93_
trityl-O-2-pyridylmethyl-D-valinol prepared above, in 80~
yield.
Step C
Preparation of N-[4-tent-butoxycarbonylamino-2,2-difluoro-
3-hydroxy-1-oxo-5-(4-benzyloxy)phenyl-pentyll-O-(2-
pyridylmethyl)-D-valinol.
Reaction Scheme A, step (b); The title compound is
prepared in a manner analogous to the procedure described
in Example 17, Step D, from the ester of Example 1, step B
and O-2-pyridylmethyl-D-valinol, prepared above, in 63$
yield, Rp=0.65 (silica gel, ethyl acetate).
Step D
Preparation of N-[4-amino-2,2-difluoro-3-hydroxy-1-oxo-5
(4-benzylaxy)phenyl-pentyl]-O-(2-pyridylmethyl)-D-valinol.
Reaction Scheme A, step (d); The title compound is
prepared in a manner analogous to the deprotection
procedure described in Example 17, step C from N-[4-tert-
butoxycarbonylamino-2,2-difluoro-3-hydroxy-1-oxo-5(4-
benzyloxy)phenyl-pentyl]-O-(2-pyridylmethyl)-D-valinol,
prepared above, in 91i: yield which is used directly in the
next step; mass spectrum, m/z 628 (MH+).
Step E
Preparation of N-(4-(N-~3-oyridylmethvl)oxvcarbonvl-L-
valvl)amino-2,2-difluoro-3-hydroxy-1-oxo-5-(4-
benzyloxy)phenyl-pentyl]-O-(2-pyridylmethyl)-D-valinol.
Reaction Scheme A', step (e); The title compound is
prepared in a manner analogous to the procedure described
in Example 17, step F from the amine prepared above and the
acid prepared is Example 8, Step A, in 53~ yield, Rg=0.14
(major) and 0.08 (minor) (silica gel,
dichloromethane/ethanol, 95/5); mass spectrum, m/z 762
(MR+).
~'1'~~1~'i
wo y6rtozav~ rcw;irsvsra7zaa
-94-
Step F
Preparation of final title camnound.
Reaction Scheme A', step (f); The final title compound
is prepared in a manner analogous to the Swern Oxidation
procedure described in Example 17, step G from the above
prepared alcohol, in 72~ yield, Rp=0.09 (silica gel, ethyl
acetate); 1H NMR (CDC13) d 8.65-8.53 (m, 1H), 7.79-7.64 (m,
2H)r 7.53-6.80 (several m, I4H), 6.60-6.10 (several m, 2H),
5.57-5.33 (m, 1H), 5.30-5.30 (m, 3H), 4.99 (s, 2H), 4.68-
4.56 (m, 2H,), 4.03-3.85 (m, 2H), 3.85-3.34 (m, 2H), 3.42-
3.24 and 2.97-2.75 (m, 2H), 2,13-1.88 (m, 2H), 1.05-0.57
(m, 12H); xsF NMR (CDC13) d (C6Fg) 50.26 (d, J=271 Hz),
49.97 (d, J=263 Hz), 49.18 (d, J=261 Hz)r 48.60 (d, J=271
Hz), 45.17 (d, J=252 H2), 45.10 (dr J=251 H2), 41.77 (d,
J=252 Hz), 40.91 (d, J=251 Hz); mass spectrum, m/z 760
(MH+).
Anal. Calcd for CalH~7N50aF2~ C, 64.81; H, 6.23; N, 9.22;
Found C, 62.91; H, 6.16; N, 8.66.
25
35
WO96If12~99
PCTIUS95/072a1
-95-
Example 21
Preparation of N-[4-(N-{2-pyridylmethyl}oxycarbonyl-L-
valyl)amino-2,2-difluoro-1,3-dioxo-5-(9-{3-
ovrzdvlmethvl}oxv)phenyl-oentyl)-O-(3-pyridylmethy1~-D-
valinol.
O ~ /N
l0
O /
N~ 0 NH CF2 NH
H ~ O
is o ~ a o
Step A
Preparation of 4-tern-butoxycarbonylamino-2,2-difluoro-3
hydroxv-5-(4-hydroxy)phenvl pentanoic acid, ethyl ester.
20 Reaction Scheme A, step (c1); A solution of 4-tert
butoxycarbonylamino-2,2-difluoro-3-hydroxy-5-[(4
benzyloxy)phenyl)pentanoic acid, ethyl ester (0.719 g, 1.5
mmol) in ethanol (50 mL) is kept for 7.5 hours under an
atmosphere of hydrogen in the presence of 10% palladium on
25 charcoal (0.079 g). The hydrogen atmosphere is then
exchanged with a nitrogen atmosphere, the suspension is
filtered and the filtrate is concentrated under vacuum to
provide the title compound (O.SOOg, 83%), Rp=0.51 (silica
gel, petroleum ether/ethyl acetate, 1/1).
Step B
Preparation of N-[4-tert-butoxvcarbonvlamino-2,2-difluoro-
3-hydroxy-1-oxo-5-(4-hydroxy)phenyl-pentyl)-O-(3-
pyridylmethvl)-D-valinol.
Reaction Scheme A, step (b); The title compound is
prepared in a manner analogous to the procedure described
in Example 19, step D, from the above prepared ester and
14'O JbIG2a94 ~ f ~ ~ ~ ~ PCTfUS9ii07241
-96-
the amine prepared in Example 17, step C, in 82% yield,
Rp=0.46 (silica gel, ethyl acetate).
Step C
Preparation of N-[4-tert-butoxycarbonylamino-2,2-difli.ioro-
3-hydroxy-1-oxo-5- 4-hydroxy)pheayl-pentyl]-O-f3-
pyridylmethyl)-D-valinol.
Reaction Scheme A, step (c2); A mixture of N-(4-tert-
butoxycarbonylamino-2,2-difluoro-3-hydroxy-1-oxo-5-(4-
hydroxy)phenyl-pentyl)-O-(3-pyridylmethyl)-D-valinol (0.376
g, 0.7 mmol, prepared above), 3-picolyl chloride HC1 (0.161
g, 0.98 mmol), cesium carbonate (0.775 q, 2.38 mmol) and
potassium iodide (0.016 g, 0.098 mmol) in anhydrous DMF (7
mi) is stirred for 66 hours under an atmosphere of
nitrogen. The reaction mixture is diluted with ethyl
acetate (50 mL) and washed with water (2 x SO mL). The
aqueous rinses are extracted with ethyl acetate (50 mL).
The combined organic extracts are dried over anhydrous
sodium sulfa e, filtered and concentrated under vacuum.
The residue is purified by flash chromatography (silica
gel, ethyl acetate/methanol, 95/5, Rf=0.25) to provide the
title compound (0.245 g, 56%): mass spectrum m,~z 629 (MH+).
Step D
Preparation of N-[4-amino-2,2-difluoro-3-hydroxy-1-oxo-5-
j4-{3-pvridvZmethvl}oxv)phenyl-pentvl]-O-(3-p~rrzdvlmethvl)-
D-valinol.
Reaction Scheme A, step (d); The title compound is
prepared in a manner analogous to the deprotection
procedure described in Example 17, steg C from N-[4-tert-
butoxycarbonylamino-2,2-difluoro-3-hydraxy-1-oxo-5-(4-
hydroxy)phenyl-pentyl]-O-(3-pyridylmethyl)-D-valinol
prepared above, in 84% yield; mass spectrum m/z 529 (MH+).
~~ dc~~~~
WO 96tti2.199 PCTN595l072a1
-97-
Steer E
Preparation of N-[4-_{2-pYridylmethyl}axycarbanyl-L-
v_alyl)amino-2 2-difluoro-3-hydroxy-1-oxo-5-(4-(3-
~ idylmethyl?oxy)phenyl-pentyl)-O-(3-pyridylmethyl)-D-
valinol.
Reaction Scheme A', step (e): The title compound is
prepared in a manner analogous to the coupling procedure
described in Example 17, step F, from the above prepared
amine and the acid prepared in Example 15, step B, in 568
yield, Rf=0.20 (silica gel, dichloromethane/ethanol, 95/5):
mass spectrum m/z 763 (MH*).
Step F
Preparation of final title compound.
Reaction Scheme A', step {f); The final title compound
is prepared in a manner analogous to the Swern Oxidation
procedure described in Example 17, step G from the above
prepared alcohol, in 708 yield, Rg=0.19 (silica gel, ethyl
acetate/acetone, 1/1); mass spectrum m/z 761 (MH+): 1H NMR
{DMSO-dg) d 9.11-8.97 (m, 1H), 8.80-8.50 (m, SH), 8.50-8.30
(m, 1H), 8.13-7.70 (m, 4H), 7.69-6.60 (m, 8H), 5.40-5.00
(m, SH), 4.71-4.49 (m, 2H), 4.10-3.80 (m, 2H), 3.80-3.51
(m, 2H), 3.51-3.30 and 2.86-2.60 (m, 2H), 2.12-1.72 (m,
2H), 1.17-0.47 (m, 12H); 19F NMR (DMSO-dg) d (CgFg) 52.91
(d, J=268 Hz), 52.11 (d, J=269 Az), 50.86 (d, J=269 Hz),
50.42 (d, J=268 Hz), 49.34 (d, J=254 Hz), 48.39 (d, J=254
Hz), 46.17 (d, J=254 Hz), 44.63 (d, J=254 Hz).
Anal. Calcd for CapHqgN607FZ: C, 63.15: H, 6.09; N, 11.05:
Found C, 62.26: H, 6.24; N, 10.76.
W09Gi02.t99 ~ ~ ~ ~ ~ ~"j PCTlLtS951072d1
-98-
Examol_e 22
Preaaration of N-(4-iN-~2-pyridy~methyl}oxycarbonyl-L-
v_alyl)amino-2 2-di~luoro-1 3-dioxo-5-(4-benzyloxv)phenvl-
pentvll-O-(2-pyridvlmethvl)-D-valinol.
n
O
O~NH
O /
O ~ O O
Step A
Pr_epa _ration of N-[4-{2-pvridvlmethyl}oxycarbonvl-L-
valyl)amino-2,2-difluoro-3-hydroxy-1-oxo-5-(4-
benzyloxy)phenyl-pentyl]-O-(2-pyridvlmethyl)-D-valinol.
Reaction Scheme A', step {e); The title compound is
prepared in a manner analogous to the coupling procedure
described in Example 17, step F from the amine prepared in
Example 20, Reaction Scheme A, step {d) and the acid
prepared in Example 15, step B, in 57~ yield, Rf=0.17
(major) and 0.12 (minor) (silica gel,
dichloromethane/ethanol, 95/5); mass spectrum mjz 762 (MH+)~
Step B
Preparation o~ ~inal title compound.
Reaction Scheme A', step {f); The final title compound
is prepared in a manner analogous to the Swern Oxidation
procedure described in Example 17, step G from the above
prepared alcohol, in 80% yield, Rf=0.22 (silica gel, ethyl
acetate); mass spectrum m/z 760 (N1H+); 1H NMR (CDC1;] 6
8.63-8.48 {m, 2H), 7.77-7.60 {m, 2H), 7.55-6.80 (several m,
13H), 6.49-6.02 (several m, 2H), 5.55-5.30 (m, 1H), 5.30-
4.95 (several m, SH), 4.72-4.52 (m, 2H), 4.05-3.82 (m, 2H),
3.82-3.35 (m, 2H), 3.42-3.23 and 2.97-2.77 (m, ZH), 2.13-
1.83 (m, 2H), 1.05-0.54 (m, 12H); 19F NMR (CDC13) d {CSFG)
50.18 (d, J=271 Hz), 49.87 (d, J=277 Hz), 49.02 (d, J=271
WO 96102499 ~ ~ ~~ ~ ~ ~ ~ PCTlU595/07241
_99_
Hz), 48.41 (d, J=271 Hz), 44.97 (d, J=252 Hzj, 44.78 (d,
J=252 Hz), 41.85 (d, J=252 Hzj, 41.12 (d, J=252 Hz).
Anal. Calcd for CQIH47N507FZr HzO; C, 63.31; H, 6.35: N,
9.00;
Found C, 63.37; H, 6.19; N, 8.85.
Example 23
Preparation of N-[4-(N-{2-pyridylmethyl}oxycarbonyl-L-
valyl)amino-2,2-difluoro-1,3-dioxo-5-(4-benzyloxy)phenyl-
pentyl]-O-methyl-D-valinol.
n
O /
,
~O~NH~ CF H
~H ~ OCH3
O ~ O O
Step A
Preparation of N-[4-{2-pyridylmethyl}oxycarbonyl-L-
valyl)amino-2,2-difluoro-3-hydroxy-1-oxo-5-(4-
benzyloxy)phenyl-pentyl]-O-methyl-D-valinol.
Reaction Scheme A', step (e); The title compound is
grepared in a manner analogous to the coupling procedure
described in Example 17, step F from the amine prepared in
Example 19, Reaction Scheme A, step (d) and the acid
prepared in Example 15, step B, in 558 yield, RE=0.23
(silica gel, ethyl acetate); mass spectrum m/z 685 (MH+j.
Anal. Calcd for CgSHqgNqOTFZ, 0.5 HZO; C, 62.32; H, 6.83; N,
8.08;
Found C, 62.36; H, 6.64; N, 7.91.
Step B
Preparation of the final title compound.
Reaction Scheme A', step (f);The final title compound
is prepared in a manner analogous to the Swern Oxidation
procedure described in Example 17, step G from the above
prepared alcohol, in 44~ yield, Rp=0.3 (silica gel,
W096Iti2a99 ~ ~ ~ ~ ~ ~ ~ PCTfUS95!(i724t
-100-
dichloromethane~'ethyl acetate, 3/7); mass spectrum mjz 683
(MHO'); 1H NMR (DMSO-d6) & 9.10-8.92 (m, 1H), 8.67-8.51 (m,
2H), 7.95-7.87 (m, 1H), 7.64-6.82 (several m. 11H), 5.30-
5.03 (m, SH), 4.10-3.93 (m, 1H), 3.93-3.78(m, 1H), 3.60-
3.37 (m,2H), 3.35-3.25 (several s, 3Hj, 3.32-3.10 and 2.87-
2.68 (m, 2H), 2.14-1.80 (m, 2H), 1.10-0.60 (m, 12H); 19F
NMR (DMSO-db) & (C6F6) 52.80 (d, J=267 Hz), 52.21 (d, J=269
Hzj, 50.93 (d, J=269 Hz), 52.21 (d, Ja269 Hz), 50.93 (d,
J=269 Ha), 49.36 (d, J=267 Hz).
Anal. Calcd for C36H4aNq07FZ, 0.5 HyO; C, 62.51; H, 6.56; N,
8.10;
Found C, 62.65; H, 6.50; N, 7.93.
Example 24
Preparation of N-{4-(N-{2-pyridylmethyi}oxycarbonyl-L-
walyl)amino-2 2-difluoro-1,3-dioxo-5-f4-benzyloxyjphenyl-
p_ent~~1]-O-benzyl-D-valinol.
n
O~NH
SOB n
O ~ i w
Step A
Preparation of N-[4-(N-{2-pvri.dylmeth~l~ oxvearbanyl-L-
valyl)amino-2 2-difluoro-3-hydroxy-1-oxo-5-(4-
3C '?benzyloxy)phenyl-pentyl]-O-benzyl-D-valinol.
Reaction Scheme A', step (e); The title compound is
prepared in a manner analogous to the coupling procedure
described in Example 17, step F from the amine prepared in
Example 5, step B and the acid prepared in Example 15, step
B in 52~; yield, Rg=0.28 (silica gel, petroleum ether/ethyl
acetate, 3/7).
Step B
Preparation of the final title compound.
21~~1~5
WO 9d102.t99 PCT/U595107241
-101-
Reaction Scheme A', step (f); The final title compound
is prepared in a manner analogous to the Swern Oxidation
procedure described in Example 17, step G from the above
prepared alcohol, in 62~ yield, Rf=0.07 (Alumina neutral Act
III, tetrahydrofuran/dichloromethane/water, 20/10/0.2),
Rf=0.27 (silica gel, ethyl acetate/petroleum ether,7/3);
mass spectrum m/z 759 (MH+); 1H NMR (CDC13) d 8.73-8.51
(m,lH), 7.79-7.65 (m, 1H), 7.42-6.45 (several m, 18H),
5.7D-5.10 (m, 4H), 5.06-4.93 (m, 2H), 4.58-4.40 (m, 2H),
4.05-3.80 (m, 2H), 3.74-3.58 and 3.55-3.40 (m, 2H), 3.35-
3.20 and 3.00-2. HD (m, 2H), 2.13-1.90 (m, 2H), 1.1D-0.50
(m, 12H); 19F NMR (CDC13) d (C6F6) 50.67 (d, J=273 Hz),
49.5D (broad s), 48.35 (d, J=273 Hz), 47.24 (d, J=255 Hz),
45.70 (d, J=255 Hz), 42.01 (d, J=255 Hz) 40.00 (d, J=255
Hz).
Anal. Calcd for Cg2HqgNq07F2, 0.5 H20: C, 65.70; H, 6.43; N.
7.30;
Found C, 65.69; H, 6.25; N, 7.19.
Example 25
Preparation of N-t4-(N-(3-pyridylmethyl}oxycarbonyl-L-
valyl)amino-2,2-difluoro-1,3-dioxo-5-(4-benzyloxy)phenvl-
pentyl]-O-(2-(2-methoxyethoxy)-1-ethyll-D-valinol.
a ~/
N
O~NH~ CF2 ,NH ~O~
H O CH3
O ~ O O
Step A
Preparation of N-trityl-0-(2-(2-methoxvethoxv)-1-ethvll-D-
valinol.
The title compound is prepared in a manner analogous to
the alkylation procedure described in Example 17, step B
from the compound prepared in Example 17, step A and 2-(2-
W0 96~(12~99 r/ ~ ~ ~ ~ ~ '~ PCTIU$J5/0'241
-102-
methoxyethoxy)ethyl-1-bromide in 8b~ yield, Rg=0.74 (silica
gel, acetone/petroleum ether, 2/8).
Step B
_Preparaticn of O-[2-(2-methoxyethoxy)-1-ethvll-D-valinol.
. A solution of N-trityl-O-[2-(2-methoxyethoxy)-1-ethyl]-
D-valinol (1.0 g, 2.28 mmol, grepared above) in dry ether
saturated with hydrogen chloride (20 mL) is kept for 2.5
hours at room temperature. The reaction is concentrated
under vacuum and the residue is purified by flash
chromatography (silica gear dichloromethane first to elute
the trityl alcohol and then dichloromethane/diethylamine.
95/5. Rg=0.20) to provide the title compound (0.46 g, 1000
as a colorless oil.
Step C
Preparation of N- 4-tert-butoxyearbonylamino-2 2-difluoro-
3 hydrpxy-1-oxo-5-(4-benzyloxy)phenyT-pentyl]-O-[2-i2-
m_ethoxyethoxy)-1-ethyl]-D-valinol.
Reaction Scheme A, step (b); The title compound is
prepared in a manner analogous to the procedure described
in Example 39. step D from the ester of Example 1, step B
and O-[2-(f-methoxyethoxy)-1-ethyl]-D-valinol, prepared
above, in 518 yield, Rg=0.37 (silica gel, petroleum
ether/ethyl acetate, 3/7).
Step D
Preparation of N-[4-amino-2 2-difluoro-3-hydroxy-1-oxo-5-
(4-benzyloxy)phenyl-pentyl]-O-[2-(2-methoxyethoxy)-1-
e_thyl]-D-valinol.
Reaction Scheme A, step (d); The title compound is
prepared in a manner analogous to the deprotection
procedure described in Example 17, step C from N-[4-tert-
butoxycarbonylamino-2,2-difluoro-3-hydroxy-1-oxo-5-(4-
benzyloxy)phenyl-pentyl]-O-(2-(2-methoxyethoxy)-1-ethyl]-D-
valinol, prepared above. in 97~ yield which is used
directly in the next step; mass spectrum m/z 539 (M,H+).
2~ y~1 ~~
1y'0 9GJ(12.i99 PCT/ITS95107241
-103-
Step E
Preparation of N-[4-(N-(3-pyridylmethyl}oxycarbonyl-L-
valyl)amino-2,2-difluoro-3-hydroxy-1-oxo-5-(4-
benzyloxy)phenyl-pentyl]-O-L2-(2-methoxyethoxy)-1-ethyl]-D-
valinol.
Reaction Scheme A', step (e); The title compound is
prepared in a manner analogous to the coupling procedure
described in Example 17, step F from the amine prepared
above and the acid prepared in Example 8, step A, in 52~
yield, Rf=0.18 (silica gel, ethyl acetate); mass spectrum
m/z 773 (MH+).
Step F
Preparation of final title compound.
Reaction Scheme A', step (f); The final title compound
is prepared in a manner analogous to the Swern Oxidation
procedure described in Example 17, step G from the above
prepared alcohol, in 76~ yield, Rg=0.09 (silica gel, ethyl
acetate); mass spectrum mjz 771 (MH'~); 1H NMR (DMSO-ds) d
9.05-8.88 (m, 1H), 8.77-8.55 (m, 2H), 8.40-8.27 (m, 1H),
7.94-7.81 (m, 1H), 7.76-6.55 (several m. 11H), 5.31-4.97
(m, 5H), 4.10-3.79 (m, ZH), 3.76-3.37 (m. 10H), 3.32 (s,
3H), 3.29-3.11 and 2.89-2.60 (m, 2H), 2.09-1.79 (m. 2H),
1.i1-0.44 (m, 12H); 19F NMR (DMSO-d6) d (C6F6) 52.97 (d,
J=267 Hzj, 52.12 (d, J=269 Hz), 50.88 (d, J=269 Hz), 49.45
(d, J=254 Hz), 45.35 (d, J=267 Hz), 48.17 (d, J=254 Hz),
46.17 (d, J=254 Hz), 44.31 (d, J=254 Hz).
Anal. Calcd for CqpHSZNaOSF2: C, 62.32; H, 6.80; N, 7.27;
Found C, 61.78; H, 6.77; N, 7.12.
~1~~~~5
wo v6rozavv rrrarsvsrD~2a~
-104-
Example 26
Pr~arationof N-[4-(N-{3-pyridvlmethyl}oxycarbonyl.-L.-,
valyl)amino-2,2-diflucro-1,3-diaxo-5-(4-{2-N-
morpholyl]ethyloxy)phenyl-pentyl]-0-methyl-D-valinol.
V
l ~ ~ \lV.~~
O
to aH Lr2 rah
OCH3
~ O O
Step A
preparation of N-(4-tent-butoxycarbonylamino-2,.2-difluoro-
~droxy-1-oxo-5-(4-hydroxy)phenyl-pentyl]-O-methyl-D-
valinol.
Reaction Scheme A, step (b); The title compound is
prepared in a manner analogous to the procedure described
in Example 17, Step D, from the ester of Example 21, step A
and the amine prepared in Example I9, step C, in 838 yield,
Rg=0.15 (silica gel, petroleum ether/ethyl acetate, 6/4);
mass spectrum m/z 46I (l~fF3+) .
Step B
Preparation of N-[4-tert-butoxycarbonylamino-2,2-difluoro-
3hydroxy-1-oxo-5-(4-(2-N-morpholvl~ethyloxy)phenvl-
pentyl]-O-methyl-D-valinol.
Reaction Scheme A, step (cy); A mixture of N-[4-tert-
butoxycarbonylamino-2,2-difluarc-3-hydroxy-1-oxo-5-(4-
hydroxy)phenyl-pentyl]-O-methyl-D-valinol (1.117 g, 2.43
mmol, prepared above), N-(2-chloroethyl)morpholine HC1
(0.633 g, 3.40 mmol), cesium carbonate (2.69 g, 8.26 mmol)
and potassium iodide (0.056 g, 0.34 moral) in dry DMF (20
mL) is stirred for 149 hours at room temperature. The
reaction mixture is then diluted with ethyl acetate (100
mL) and washed with brine (2 x 1D0 mL). The aqueous washes
are extracted with ethyl acetate (100 mL). The combined
organic extracts are dried over anhydrous sodium sulfate,
W09G/02449 ~ ~ ~ ) ~ '~' ~ PCT/US95107241
-105-
filtered and concentrated under vacuum. The residue is
purified by flash chromatography (silica gel, ethyl
acetate, Rp=O.i8) to provide the title compound (0.993 g,
71~;).
Step C
Preparation of N-[4-amino-2,2-difluoro-3-hydroxy-1-oxo-5-
-(2-N-morpholyl}ethyloxylphenvl-pentvl]-O-methyl-D-
valinol.
Reaction Scheme A, step (d); The title compound is
prepared in a manner analogous to the deprotection
procedure described in Example 17, step C from N-[4-tert-
butoxycarbonylamino-2,2-difluoro-3-hydroxy-1-oxo-5-(4-{2-N-
i5 morpholyl}ethyloxy)phenyl-pentyl]-O-methyl-D-valinol,
prepared above, in 858 yield which is used directly in the
next step.
Step D
Preparation of N-[4-(N-{3-pvridvlmethvl}oxvcarbonvl-L-
valyl)amino-2,2-difluoro-3-hydroxv-1-oxo-5-(4-{2-N
morpholyl}ethylaxy)phenyl-pentyll-O-methyl-D-valinol.
Reaction Scheme A', step (e); The title compound is
prepared in a manner analogous to the procedure described
in Example 17, Step F from the amine prepared above and the
acid prepared in Example 8, step A, in 80$ yield, Rg=0.19
(silica gel, ethyl acetate/methanol, 9/1); mass spectrum
m/z 708 (MH+).
Step E
Preparation of final title compound.
Reaction Scheme A', step (f); A mixture of the above
prepared alcohol (0.280 g, 0.396 mmol), 1,1,1-triacetoxy-
1,1-dihydra-1,2-benziodoxol-3(lAy-one (0.672 g, 1.583 mmol,
Dess-Martin periodinane) and tert-butanol (0.075 mL, 0.792
mmol) in freshly distilled dichloromethane (10 mL,
distilled over Py05) is stirred for 15 minutes at room
temperature. The reaction mixture is then hydrolyzed with
-,
W'0 9l>102 f39 ~ ~ ~ ~ ~ ~ PC'ItUSJ5li172d1
-106-
isoprapanol (1.2 mL) and concentrated under vacuus~. The
residue is suspended in dichioromethane (3 mL), filtered
through a Flurapore filter which is rinsed with
dichloromethane (2 x I mL). The filtrate is concentrated
under vacuum and the residue purified by flash
chromotagraghy (2x, silica gel, dichloramethane%methanal,
99J1 to remove the by-products of the Dess-Martin Reagent,
then 98/2 and 96/4 to elute the title compound) to provide
the title compound (0.112 g, 28~), Rf=0.11
(dichloromethane/methanol, 95/5); mass spectrum m/z 706
(MH+); 1H NMR (DMSO-d6) 8 8.99-8.70 (m, 1H), 8.64-8.43 (m,
3H), 7.82-7.73 (m, 1H), 7.47-6.70 (several m, 6H), 5.16-
4.92 (m, 3H), 4.08-3.95 (broad s, 2H)r 3.95-3.71 (m, 1H},
3.71-3.65 (m, 1H), 3.60-3.48 (broad s, 4H), 3.43-3.34 (m,
2H), 3.22-3.18 (2s, 3H}, 3.24-3.03 and 2.77-2.56 (m, 2H),
2.54-2.34 (m, 4A), 2.02-1.70 (m, 2H), 1.10-0.50 (m, 12H);
1gF NMR (DMSO-d5) d (CSFg) 52.87 (d, J=267 Hz}, 52.22 (d,
J=269 Hz), 50.91 (d, J=269 Hz), 49.38 (d, J=267 Hz).
Anal. CalCd ~ar CgSHqgN50gF2~0.5 H20; Cr 58.81; H, 7.05; N,
9.80;
Found C, 58.61; H, 6.88; N, 9.93.
30
2~~~1~5
W O 9G!(12499 PCTNS95l07241
-107-
In a further embodiment the present invention provides
a method of treating a patient afflicted with a viral
infection comprising the administration thereto of an
effective antiviral amount of a compound of formula (I).
. The term "viral infection" as used herein refers to an
abnormal state or condition characterized by viral
transformation of cells, viral replication and
proliferation. Viral infections for which treatment with a
compound of formula (I) will be particularly useful include
retroviruses such as but not limited to HTLV-I, HTLV-II,
HTLV-III (HIV virus), murine leukemia virus, feline
leukemia virus, cytomegalovirus(CMV), avian sarcoma virus
and the like. In addition treatment with a compound of
formula (I) would be useful in treating a wide range of
states of, HIV infection: AIDS, AAC (AIDS related complex),
both symptomatic and asymptomatic, and actual or potential
exposure to HIV. For example, the compounds of this
invention are useful in preventing infection by HIV after
suspected past exposure to HIV by, e.g., blood transfusion,
accidental needle stick, or exposure to patient blood
during surgery.
An "effective antiviral amount" of a compound of
formula (I) refers to an amount which is effective, upon
single or multiple dose administration to the patient, in
controlling the growth of the virus or in prolonging the
survivability of the patient beyond that expected in the
absence of such treatment. As used herein "controlling a
viral infection" refers to slowing, interrupting, arresting
or stopping the viral transformation of cells or the
replication and proliferation of the virus and does not
necessarily indicate a total elimination of the virus.
The present invention further provides a method of
inhibiting HIV protease in a patient in need thereof
2~'~~1~~
Vf0 96~'(t2~99 PCT/II595f071:11
-108-
comprising administering to said patient an effective
inhibitory amount of a compound of formula (T).
It is understood that patients suffering from a
retrovirus, such as HTLV-III are in need of an HIV protease
inhibitor such as a compound of formula (I).
As used herein, the term "gatient" refers to a warm-
blaoded animal, such as a mammal, which is afflicted with
a particular viral infection. It is understood that
humans, mice and rats are included within the scope of the
term "patient".
Administration of a compound of formula (I) to a
patient results in inhibition of HIV protease in the
patient. Thus, by treatment of a patient with a compound
of formula (I) retroviruses, such as HTLV-III, are
inhibited or suppressed.
A patient is in need of treatment with an agent which
inhibits HIV protease, such as a compound of formula (i),
where the patient is suffering from certain viral
infections for which HiV protease is implicated as a.
contributing factor in the progression of the disease.
Based on standard clinical and laboratory tests and
procedures, an attending diagnostician, as a person skilled
in the art, can readily identify those patients who are in
need of trea ment with an agent which inhibits HIV
protease, such as a compound of formula (I).
An "effective inhibitory amount" of a compound of
formula (I) is that amount which is effective, upon single
or multiple does administration to a patient, in providing
an inhibition of HIV protease.
21'~~' ~5
WO 961t12~99 PCTIUS951U7241
-i09-
As used herein the term "effective amount" refers to an
effective antiviral or inhibitory amount of a compound of
formula (I). An effective amount can be readily determined
by the attending diagnostician. as one skilled in the arty
by the use of known techniques and by observing results
obtained under analogous circumstances. In determining the
effective amount or dose, a number of factors are
considerec by the attending diagnostician, including, but
not limited to: the species of mammal; its size, age, and
general health; the specific viral infection involved; the
degree of or involvement or the severity of the viral
infection; the response of the individual patient; the
particular compound administered; the mode of
administration; the bioavailability characteristics of the
preparation administered; the dose regimen selected; the
use of concomitant medication; and other relevant
circumstances.
An effective amount of a compound of formula (I) is
expected to vary from about 0.1 milligram per kilogram of
body weight per day (mg/kg/day) to about 100 mg/kg/day.
Preferred amounts are expected to vary from about 0.5 to
about 10 mg/kg/day.
In effecting treatment of a patient afflicted with a
viral infection, a compound of formula (I) can be
administered in any form or mode which makes the compound
bioavailable in effective amounts, including oral and
parenteral routes. For example, compounds of formula (I)
3Q can be administered orally, subcutaneously,
intramuscularly, intravenously, transdermally,
intranasally, rectally, and the like. Oral administration
is generally preferred. One skilled in the art of
preparing formulations can readily select the proper form
and mode of administration depending upon the particular
characteristics of the compound selected, the viral
wo ~croza9y rcr~trs~sro~zai
-1I0-
infection to be treated, the stage of the infection, and
other relevant circumstances.
The compounds of formula (I) can be administered alone .
S or in the form of a pharmaceutical composition in
combination with pharmaceutically acceptable caeeiers or
excipients, the proportion and nature of which are
determined by the solubility and chemical properties of the
compound selected, the chosen route of administration, and
standard pharmaceutical practice. The compounds of the
invention, while effective themselves, may be formulated
and administered in the form of their pharmaceutically
acceptable salts for purposes of stability, convenience of
crystallization, increased solubility and the like.
In another embodiment, the present invention provides
compositions comprising a compound of formula (I) in
adatixture or otherwise in association with one or more
inert carriers. These compositions are useful, for
example, as assay standards, as convenient means of making
bulk shipments, oz as pharmaceutical co-~positions. An
assayable amount of a compound of formula (I) is an amount
which is readily measurable by standard assay procedures
and techniques as are well known and appreciated by those
skilled in the art. Assayable amounts of a compound of
formula (I) will generally vary from about 0.001% to about
75% of the composition by weight. Inert carriers can be
any material which does not degrade or otherwise covalently
react with a compound of formula (I). Examples of suitable
inert carriers are water; aqueous buffers, such as those
which are generally useful in high Performance Liquid
Chromatography (HPLC) analysis; organic solvents, such as
acetonitrile, ethyl acetate, hexane and the like; and
pharmaceutically acceptable carriers or excipients.
More particularly, the present invention provides
pharmaceutical compositions comprising a
WO 96102x99 ~ ~ ~ ,J ~ 4 .~ PCTlUS9g107241
-111-
therapeutically effective amount of a compound of
formula (I) in admixture or otherwise in association
with one or more pharmaceutically acceptable carriers
or excipients.
The pharmaceutical compositions are prepared in a
manner well known in the pharmaceutical art. The
carrier or excipient may be a solid, semi-solid, or
liquid material which can serve as a vehicle or medium
for the active ingredient. Suitable carriers or
excipients are well known in the art. The
pharmaceutical composition may be adapted for oral or
parenteral use and may be administered to the patient
in the form of tablets, capsules, suppositories,
'-5 solution, suspensions, or the like.
The compounds of the present invention may be
administered orally, for example, with an inert diluent
or with an edible carrier. They may be enclosed in
gelatin capsules or compressed into tablets. For the
purpose of oral therapeutic administration, the
compounds may be incorporated with excipients and used
in the form of tablets, troches, capsules, elixirs,
suspensions, syrups, wafers, chewing gums and the like.
These preparations should contain'at least 4% of the
compound of the invention, the active ingredient, but
may be varied depending upon the particular form and
may conveniently be between 4$ to about 70~ of the
weight of the unit. The amount of the compound present
in compositions is such that a suitable dosage will be
obtained. Preferred compositions and preparations
according to the present invention are prepared so that
an oral dosage unit form contains between 5.0-300
milligrams of a compound of the invention.
The tablets, pills, capsules, troches and the like
may also contain one or more of the following
C "'
WO 96/02199 ~ ~ ~ .J ~ cr. ~ PCT/IS995/072d1
-112-
adjuvants: binders such as microcryscalline cellulose,
gum tragacanth or gelatin; excigients such as starch or
lactose, disintegrating agents such as alginic acid,
Primogel, corn starch and the like; lubricants such as ,
magnesium stearate or sterotex; glidants such as
colloidal silicon dioxide; and sweetening agents such.
as sucrose or saccharin may be added or a flavoring
agent such as peppermint, methg~l saiicylate or ozange
flavoring. When the dosage unit form is a capsule, it
may contain, in addition to materials of the above
type, a liquid carrier such as polyethylene glycol or a
fatty oil. Other dosage unit forms may contain other
various materials which modify the physical form of the
dosage unit, for example, as coatings. Thus, tablets
I5 or pills may be coated with sugar, shellac, or other
enteric coating agents. A syrup may contain, in
addition to the present compounds, sucrose as a
sweetening agent and certain preservatives, dyes and
colorings and flavors. Materials used in preparing
these various compositions should be pharmaceutically
pure and non-toxic in the amounts used.
For the purpose of parenteral therapeutic
administration, the compounds of the present invention
may be incorporated into a solution or suspension.
These preparations should contain at least 0.1~ of a
compound of the invention, but may be varied to be
between 0.1 and about 50% of the weight thereof. The
amount of the inventive compound present in such
compositions is such that a suitable dosage will be
obtained. Preferred compositions and preparations
according to the present invention are prepared so that
a parenterai dosage unit contains between 5.0 to 100
milligrams of the compound of the invention.
The solutions or suspensions may also include the
one or more of the following adjuvants: sterile
WO 96102499
PCTIU595/07241
-113-
diluents such as water for injection, saline solution,
fixed oils, polyethylene glycols, glycerine, propylene
glycol or other synthetic solvents; antibacterial
agents such as benzyl alcohol cr methyl paraben;
antioxidants such as ascorbic acid or sodium bisulfite;
chelating agents such as ethylene diaminetetraacetic
acid; buffers such as acetates, citrates or phosphates
and agents for the adjustment of tonicity such as
sodium chloride or dextrose. The parenteral
preparation can be enclosed in ampules, disposable
syringes or multiple dose vials made of glass or
plastic.
The present invention is also directed to combinations
of the HIV protease-inhibitory compounds with one or more
agents useful in the treatment of AIDS, such as, for
example, with known antiviral agents suitable for treating
HIV 1 and HIV 2 viral infections, e.g., AZT, with or
without a PNPase inhibitor, or in conjunctive therapy with
DDI and a PNPase inhibitor.
30
W0 9GI0~499 ~ 9 ~ ~ ~ "~ ~ FC'I'fLTS4,~s107Z~11
oL
-I14-
The compounds of this invention may be assayed fc-
their HIV-protease inhibition using the following published
techniques.
Preparatian of Retroviral Enzyme
and
Assay for Inhibition of the Prctease
A) Preparation of Retroviral Enzyme
IO To prepare the recombinant protease, the HIV protease
is expressed via E. Coli by the published work of
C. Guenet, et ai., in European Sournal of Pharmacology.
Molecular Pharmacology Section, i72 (1989) 443-451.
B) Assay for inhibition of Recombinant Viral Protease
Inhibition of the reaction of the protease with a
peptide substrate [Ser-Gln-Asn-Tyr-Pro-Ile-Val-NH2,
Km = 1 mM] are in 50 mM Na acetate, 10~ glycerol, 5~
ethyleneglycol, pH 5.5, at 37°C for 1 hour. Various
concentrations of inhibitor in 10 ~1 DMSO are added to
80 u1 of assay soiutian and the reaction is initiated
by the addition of 10 u1 (1.& pp) of recombinant
protease. The reaction is quenched with I6 y~l of 4 M
perchloric acid. Products of the reaction are separated
by HPLC (VYDAC wide pore 5 cm C-18 reverse phase,
acetonit-ile gradient, 0.1~ trifluoroacetic acid). The
extent of inhibi=ion of the reaction is determined from
the peak heights of the products. HPLC of the products,
independently synthesized, provide quantitation
standards and confirmation of the product composition.
By following the techniques referenced above, as well
as by utilization of other known techniques, as well as by
comparison with compounds known to be useful for treatment
of the above-mentioned disease states, it is believed that
adequate material is available to enable one of ordinary
skill in the art to practice the invention.
WO 9G!02499 ~ ~ ~ ~ 1 L ~ PCT/US95/07241
-115-
As is true for most classes of compounds found to be
useful in the pharmceutical industry, certain subgeneric
groups and certain specific compounds are more preferred
such as those exemplified and shown in the following
charts.
15
25
35
R'O 9b;02.199 ~ ~ ~ ~ ~ ~ ~ PCTIUS95l(172d1
_ 116 -
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x
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WO 96t02~99 ~ ~ ~, °~ ~ L PCTIUS95I07241
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W09bIf12S99 ~ ~ ~ ~ ', ~ PCT.iiIS9SJ072aI
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W0 96~'U2499 PCTJLrS95l(i7241
-122-
The following list illustrates compounds according to
the present invention;
1, a,a-difluoro-Y-[((2-(Rj-
[I(hydroxyjphenylacetyl]amino}-3-methyl-1-oxobutyl]amino]-
6-oxo-4-(phenylmethoxy}-N-(phenylmethylj-benzene-
pentanamide;
2. [fi5-(6R*, 9R*, 138*)]-4,4,-di~luoro-9-(1-
1D methylethylj-3,5,8,11-tetraoxo-1,13-diphenyi-6-[(4-
(phenylmethoxy)phenyl]methyl]-12-oxa-2,1,10-
triazatetradecan-14-oic acid, methyl ester;
3. a,a-difluora-y-[[3-methyl-I-oxo-2-[((1-oxo-trans-3-
1; phenyl-2-propenyl)amino]butyljamino]-9-oxo-4-
(phenylmethoxyj-N-(phenylmethyl)-benaene-pentanamide;
4. a,a-difluoro-Y-[[2-(2-hydroxy-1-oxo-4-
(ghenylbutyl)amino]-3-methyl-1-oxobutyl]amino]-~-oxo-4-
20 (phenylmethoxyj-N-(phenylmethyl)-benzene-pentanamide;
5. N-[1-[[[3,3-difluoro-4-[[2-methyl-1-
[(phenylmethoxy)methyl]-propyl]amino]-2,4-dioxo-1-([4-
(phenylmethoxy)phenyl]-methyl]butyl]amino]carbonyl]-2-
25 methylpropyl]-g-oxo-4-morpholinepropanamide;
6. [1R-(1R*,25*)]-a, a-difluoro-y-((2-(R)-
[({hydroxy)phenylacetyl]-amino]-3-methyl-1-oxobutyl]amino]-
N-[2-methyl-1-((phenylmethoxy)methyl]propyl]-3-oxa-4-
30 (phenylmethoxy)-benzenepentanamide;
7. N-[1-([[3,3-difluoro-4-([2-methyl-1-
[(phenylmethoxy)-methyl]-propyl]amino]-2,4-dioxo-1-[[4-
(phenylmethoxy)phenyl]- methyl]-butyl]amino]carbonyl]-2-
35 methylpropyl]-6-{4-morpholinyi-carbonyl)-3-
pyridinecarboxamide;
WO 9G!02499 ~~ ~ PCTIUS95/072a1
-123-
8. 7.7-difluaro-4,12-bis(1-methylethyl)-6.8,11-trioxo-
1-phenyl-9-((4-(phenylmethoxy)phenyl]methyl)-2-oxa-5,10,13-
triazatetradecan-14-oic acid, 3-pyridinylmethyl ester;
9. [1R-(1R*,2S*)]-a,a-difluoro-y-[(3-methyl-2-([4-(4-
morphalinylsulfonyl)benzoyl]amino]-1-oxabutyl]amino]-N-[2-
methyl-i-[(phenylmethoxy)methyl]propyl]-8-oxo-4-
(phenylmethoxy)-benzenepentanamide;
Z0. (1R-(1R*,2S*)]-a,a-difluoro-y-[[3-methyl-2-[[4-(4-
morpholinylcarbonyl)benzoyl]amino]-1-oxobutyl]amino]-N-[2-
methyl-1-[(phenylmethoxy)methyl]propyl]-g-oxa-4-
(phenylmethoxy)-benzenepentanamide;
11. N-[1-[([3,3-difluoro-4-[(2-methyl-1-
[(phenylmethoxy)methyl)-propyl]amino]-2,4-dioxo-1-[(4-
(phenylmethoxy)phenyl]-methyl]butyl]amino]carbonyl]-2-
methylpropyl]-9-morphalineacetamide;
12. N-[1-[[[3,3-difluoro-4-[[2-methyl-1-
[{phenyimethoxy)methyl]-propyl]amino]-2,4-dioxo-1-[[4-
(phenylmethoxy)phenyl]methyl]-butyl]amino]carbonyl]-2-
methylprapyl]-1H-imidazole-1-acetamide;
13. [1-[[[3,3-difluoro-2,4-diaxo-1-[[4-
(phenylmethoxy)phenyl]-methyll-4-[(2-
pyridinylmethyl)amino]butyl]amino]carbonyl]- 2-
methylpropyl]-carbamic acid, 3-pyridinylmethyl ester;
35
W0961(i2-t99 ~ ~ ~ ~ ~ ~~ ~ PCTlUS9~5107241
-124-
14. (i-(([3,3-difluoro-2,4-dioxo-1-[[4-
1S
(phenylmethoxy)phenyl]-methyl]-4-[(3-
pyridinylmethyl)amino]butyl]amino]carbonyll-2-
methylpropyl]-carbamic acid, 3-pyridinylmethyl ester;
5
15. [1-(CL3,3-difluoro-2,4-dioxo-1-[[4-(phenylmethoxy)-
phenyl]-methyl]-4-((2-pyridinylmethyl)amino]butyl]-
ami.no]carbonyl]- 2-methylpropyl]-carbamic acid, 2-
pyridinylmethyl ester;
16. [1-[[[3,3-difluoro-2,4-dioxo-1-[(4-(phenylmethoxy)-
phenyl]-methyl]-4-[(3-pyridinylmethyl)amino]butyl]-
amino]carbonyl]- 2-methylpropyl]-carbamic acid, 2-
pyridinylmethyl ester;
17. N-[4-(N-{2-pyridylmethyl}oxycarbonyl-L-valyl)amino-
2,2-difluoro-I,3-dioxo-5-(4-benzyloxy)phenyl-pentyl]-O-{3-
gyridylmethyl)-D-valinol;
18, N-[4-(N-{3-pyridylmethyl}oxycarbonyl-L-valyl)amino-
2,2-difluoro-1,3-dioxo-5-(4-benzyloxy)phenyl-pentyl]-O-(3-
pyridylmethyl)-D-ualinol;
19. N-[4-(N-{3-pyridylmethyl}oxycarbonyl-L-valyl)amino-
2,2-difluoro-1,3-dioxo-5-(4-benzyloxy)phenyl-pentyl]-O-
methyl-D-valinol;
20. N-{4-(N-{3-pyridylmethyl}oxycarbonyl-L-valyl)amino-
2,2-difluoro-1,3-dioxo-5-{4-benzyloxy)phenyl-pentyl]-O-(2-
pyridylmethyl)-D-valinol;
21. N-I4-(N-{2-pyridylmethyl}oxycarbonyl-L-
va?p_)amino-2,2-difluoro-1,3-dioxo-5-(4-{3-
pyridylmethyl}oxy)phenyl-pentyi]-O-(3-pyridylmethyi)-D-
3S valinol;
R'O 96!02a99 ~ ~ ~ r} ~ ,~ ~ PCTlUS95/07241
-125-
22. N-[4-(N-{2-pyridylmethyl}oxycarbonyl-L-valyl)amino-
2,2-difluoro-1,3-dioxo-5-(4-benzyloxy)phenyl-pentylj-O-(2-
pyridylmethyl)-D-valinol;
23. N-[4-(N-{2-pyridylmethyl}oxycarbonyl-L-
valyl)amino-2,2-difluoro-1,3-dioxo-5-(4-benzyloxy)phenyl-
pentylJ-0-methyl-D-valinol;
24. N-{4-(N-{2-pyridylmethyl}oxycarbonyl-L-valyl)amino-
2,2-difluoro-1,3-dioxo-5-(4-benzyloxy)phenyl-pentylj-O-
benzyl-D-valinol;
25. N-{4-(N-{3-pyridylmethyl}oxycarbonyl-L-valyl)amino-
IS 2,2-difluoro-1,3-diaxa-5-(4-benzyloxy)phenyl-pentyl)-O-[2-
(2-methoxyethoxy)-1-ethyl)-D-valinol; and
26. N-[4-(N-{3-pyridylmethyl}axycarbonyl-L-valyl)amino
2,2-difluoro-1,3-dioxo-5-(4-{2-N-morpholyl]ethyloxy)phenyl
gentylj-O-methyl-D-valinol.
30
