Note: Descriptions are shown in the official language in which they were submitted.
HA375a
-1- 1310792
N-HETEPcOCYCLIC ALCOHOL RENIN INHIBITORS
.... . _ .
Jones et al. in WO 84/03044 disclose renin
inhibiting tetra-, penta-, or hexapeptide
analogues of the formula
X-D-E-A-B-Z-W
where X and W are terminal groups; D, E, B and Z,
of which any one or, except with reduced
analogues, two may be absent, are aromatic, lipo-
philic or (in the case of E) aromatic, lipophilic,
or basic amino acid or amino acid analogue
residues, and ~ is an analogue of a lipophilic or
aromatic dipeptide residue wherein the peptide link
is replaced by one to four-atom carbon or carbon-
nitrogen link which as such or in hydrated form is
an unhydrolyzable tetrahedral analogue of the
transition state of the peptide bond as given
above. In particular, A is defined as
R4 Rl R2
-N - C - M C -
R5 R6
~'
~A375a
-2~
1 3 1 079~
wherein M can b~ -C~O~.
Sz~lke et al. in European Patent ~pplication
104,041, published in March 1984, disclose renin
inhibitory polypeptides including the partial sequence
S
X -A- B- Z- W and
X-Ph~-Hi8-A-~-Z~W
10 wherein A i~ Rl R3 R2 O.
-Ng-C~- G-~ C~- C-
and G is
OE~
-C8 -C~2
X is hydkog~n, protecting g~oup, or an amino acyl
residue, B i~ a lipop~ilic ami~o acyl r~sidu~, and
Z plu~ W ar~ an a~ino alcohol r~idu~ or Z i~
a~inoacyl and W is h~dro~y, e~t~r, amido, ~tc.
Mat~u~da et al. in U.S. Patent 4,54~,926
disclo~s renin inhibiting peptido~ of th~ for~ula
N~
O C~ O Bu~
~ 2 1l 1
R - C- NH - C~ - C ~N~ C~-X
HA375a
-3-
1 3 t 07q2
wherein But represents an isobutyl or sec-butyl
group and X includes a group of the formula
-CH(R2 )_y
Gordon et al. in U.S. Patent 4,514,391
disclose hydroxy substituted peptide compounds of
the formula
OH R Rl o
R3- CH -CH - CH2- N -CH - C -X
NH
C~O ,
R2
which possess angiotensin converting enzyme or
lS enkephalinase inhibition activity.
~ his invention is directed to new hetelocyclic
alcohol containing renin inhibitors of formula I
including pharmaceutically acceptable salts thereof
~I)
R5 O R4 R3
ll l 11 1
~5 X ~ NH- CH - C ~ NH -CH - C -NH -CH -CH - R
0~
O O
Il ~ 11
3~ X is R6-~CH2)m~, R6-(CH2)m-C N CH C ,
Rlo o
Il l 11 11
R6-(CH2)m-C-t NH ~CH~ C ~--q , R6-~CH2)m-O-C-,
_4~ HA375a
13107Q~
o o
Il 11
R6--(CH2)n~C- , R6-(CH2)m~ C-
O
Il
CN _ C , R6~(CH2)m-S2- or
o ,,
R6- ( CEI2 )m-CH--c-
(CH2)m,
R3, R4, R5 and R1o are independently selected
from hydrogen, lower alkyl, halo substituted lower
alkyl, -(CH2)n-aryl, -(CH2)n-heterocyclo, -(CH2)n-OH,
-(CH2)n-O-lower alkyl, -(CH2)n-NH2, -(CH2)n-SH,
-(CH2)n-S-lower alkyl, ~(CH2)n~~(CH2)g~OH,
( 2)n (CH2)g~NH2~ ~(CH2~n~S-(CH2) -OH~
ll
-(CH2)n-C-OH, ~(CH2)n~S~(CH2)g~NH2,
N~
30 -(CH2)n-NH-C , ~.
NH2
_5_ ~375a
1 3 1 07~2
o
Il
( 2~n C NH2 ~ (CH~)n ~ J ~ I-(CH2)
R8
and -(CH2~n-cycloalkyl~
R6 and R6' are independently selected from
lower alkyl, cycloalkyl, aryl and heterocyclo.
p is zero or one.
g is zero or one.
m and m' are independently selected from5 zero and an integer from 1 to 5.
n is an integer from 1 to 5.
g is an integer from 2 to 5.
7 is CH2 O CH2 ~ or -CH2 ~ .
R8 is 2,4-dinitrophenyl, -C-O-C~2 ~ ,
-52 ~ CH3 ~ or -CH2 O-CH
~0
-6- HA375a
1 3 1 0792
Rl is a fully saturated, partially saturated,
or unsaturated monocyclic N-heterocyclic ring of 5
or 6 atoms containing at least one N atom or a
~icyclic ring in which such N-heterocyclic ring is
fused to a benzene rin~. The N-heterocyclic ring
can also include an O or S atom or up to three
additional N atoms. The N-heterocyclic ring is
attached to -CH- by way of an available carbon
OH
atom. An available N atom in the N-heterocyclic
ring can be substituted with an N-protecting group-
such as -CH2-0-CH2 ~ , -S2 ~ H3 ~
or 2,4-dinitrophenyl, or lower alkyl, -(CH2)n ~ ,
or -(CH2)n-cycloalkyl. Similarly, an available C
atom in the monocyclic ring or an avaliable C atom
in the benzene portion of the bicyclic ring can be
substituted with lower alkyl,
~(CH2)n~> '
or -(CH2)n-cycloalkyl.
Preferred N-heterocyclic rings are
lR2 lR2
~ ~ Rg, N ~ 9 ' N ~ 9
R
r~ ~ 2
R2
Rg
HA375a
~7 1 3 1 0792
~S ~o
N $1 N ~J
R~ ~'<9
R9 ~ N ~ Rg ' --N~ ~ 9
R2
1 2
_~Rg , ~ Rg ~ ~Rg
T R~ and ~N~ Rg -
2 5 CH2 0 CH2~ , -S02~ c~3
2,4-dinitrophenyl, hydrogen, lower alkyl, -(CH2)
or -(CH2)n-cycloalkyl.
Rg is hydrogen, lower alkyl,
-(CH2)
or -(CH2)n-cycloalkyl.
-8- HA375a
~ 3 1 0 7 9 2
,--
~ N- represents a heterocyclic ring of the
S ~ormula ~ (CH2)~\
Y N-
(CH~)b
wherein Y is -CH2, O, S, or N-Rg, a is an integer
from 1 to 4, and b is an integer from 1 to 4
provided that the sum of a plus b is an integer ~rom
2 to 5 and such heterocyclic rings wherein one carbon
atom has a substitu~nt selected from lower alkyl,
lower alkoxy, lower alkylthio, halo, CF3, nitro, or
~ydroxy.
~0 This invention in its broadest aspects
relates to the compounds of formula I above, to
compositions and the method of using such compounds
as antihypertensive agents, and intermediates useful
in the preparation of such compounds.
The term lower alkyl used in defining
various symbols refers to straight or branched
chain radicals having up to seven carbons.
Similarly, the terms lower alkoxy and lower
alkylthio refer to such lower alkyl groups
attached to an oxygen or sulfur. ~he preferred
lowar alkyl groups are straight or branched chain
o~ 1 to 5 carbons.
_g_ HA375a
1 31 07~2
The term cycloalkyl refers to saturated
rings of 4 to 7 carbon atoms with cyclopen~yl and
cyclohexyl being most preferred.
The term halogen refers to chloro, bromo and
fluoro.
The term halo substituted lower alkyl refers
to such lower alkyl groups described above in
which one or more hydrogens have been replaced by
chloro, bromo or fluoro groups such as trifluoro-
methyl, which is preferred, pentafluoroethyl,2,2,2-trichloroethyl, chloromethyl, bromomethyl, --
etc.
The term aryl refers to phenyl, l-naphthyl,
2-naphthyl, mono substituted phenyl, l-naphthyl,
lS or 2-naphthyl wherein said substituen~ is lower
alkyl of 1 to 4 carbons, lower alkylthio of 1 to 4
carbons, lower alkoxy of 1 to 4 carbons, halogen,
hydroxy, amino, -NH-alkyl wherein alkyl is of 1 to
4 carbons, or -N(alkyl)2 wherein alkyl is of 1 to 4
carbons, di or tri substituted phenyl, l-naphthyl
or 2-naphthyl wherein said substituents are
selected from methyl, methoxy, methylthio,
halo~en, and hydroxy.
The term heterocyclo refers to fully saturated
or unsatuxated rings of 5 or 6 atoms containing one
or two 0 and S atoms and/or-one to four N atoms
provided that the total number of hetero atoms in
the ring is 4 or less. The hetero ring is
attached by way o an available carbon atom.
Preferred hetero groups include 2- and 3-thienyl,
2- and 3-furyl, 2-, 3- and 4-pyridyl. The term
hetero also includes bicyclic rings wherein
HA375a
-lO- I 3 1 0792
the five or six membered ring containing 0,
s and N atoms as defined above is fused to a
benzene ring. The preferred bicyclic ring
is benzimida201yl.
The compounds of formula I wherein
o
Il
6 ~ H2)m C
can be prepared by coupling an alcohol of the formula
(II)
H2N-CH -CH-R
OH
with a peptide of the formula
~III)
200 R5 R4
R6-(CH2)m~O~C-~ N~ -CH - C~-p NH- CH - COOH
This reaction is preferably performed in a solvent
such as dimethylformamide and in the presence of
hydroxybenzotria201e, N-methylmorpholine, and a
coupling agent such as dicyclohexylcarbodiimide.
The corresponding compounds of formula I
wherein p is zero can be prepared by coupling
the alcohol of formula II with the amino acid of
the formula
-11- HA375a
1 3 1 0 ~q~
(IV~
O R4
Il I
R6-(cH2)m-o-c-NH-cH -COOH
to yield the products of the formula
(V)
Rl R3
Il l 11 1
R6(C~)m-O-C-NH-CH-C -NH-CH -CH-R
OH
When R6-(CH2)m- is t-butyl or benzyl,
lS then t~e product of formula V can be treated
so as to remove the t-butoxycarbonyl or
ben2yloxycarbonyl group such as by the use
of anhydrous hydrochloric acid when R6 is t-butyl
to yield the amine of the formula
(VI)
R4 R3
Il
H2N-C~--C-- N~--CH--CH-Rl
I
~5 OH
Coupling with the acylated amino acid of the formula
(VII)
O R
~.
R6-(CH2) -O-C -NH -CH -COOH
yialds ~he products of formula I wherein
p iS one.
HA375a
-12-
~ 3 1 ~7q2
The compounds of formula I wherein X is
o
Il
other than R6-(CH2)m-O-C-
s can b~ prepared by treating the product of
formula I wherein X is
O o
-C -o-c(cH3)3 or -C O C~2 ~
to remove the t-butoxycarbonyl or benzyloxycarbonyl
group and yield the intermediates of the formula _-
(VIII)
R5 1I R4 11 l3
H2N-CH--C--NH--CH--C--N~--CH~ CH-R
OH
The amine of form~la VIII or formula VI is treated
with the halide of the formula
(IX)
R6- ( CH2 )m-halo
particularly where halo is Br to give the products
of formula I wherein X is R6-(CH2)m~.
~he compounds of formula I wherein X is
o
R6-O-(CH2)n-C- can be prepared by treating ~le
amine of formula VIII or VI with the acid chloride
1 3 1 0792
-13- HA375a
of the formula
(X)
o
Il
~6 (CH2)n-c-Cl
in the presence of triethylamine.
The compounds of formula I wherein X is
R6-(CH2)m-S02- can be prepared by treating the
amine of formula VIII or VI with the substituted
sulfonyl chloride of the formula
(XI)
6 ( 2)m 2
~he compounds of formula I wherein X is
o
Il
R6-(CH2)m-C- can be prepared by treating the
amine of formula VIII or VI with the acid chloride
of the formula
(XII)
O
Il
R6- ( CH2 )m-C-Cl
~5
in ~he presence of triethylamine. Alternatively,
these compounds can also be prepared by coupling
the carboxylic acid of the fo~ula
(XIII)
3~ 0
R6-(CH2)m-C-OH
~ 3 1 0792
HA375a
to the amine of formula VI or VIII in the presence
of a coupling agent sucn as dicyclohexylcarbo~
diimide and 1-hydroxybenzotriazole hydrate.
~he compounds of formula I wherein X is
Rl o O
Il l 11
R6- ( CH2 )m~C~ (NH-CH--C ~
and q is one can be prepared by acylating the
amino acid of the formula
(XIV) -.
RllO
H2~CH - C - OH
with the acid chloride of formula XII in the
presence of base such as sodium hydroxide (i.e. at
a pH of about 8) and in a solvent such as tetra-
hydrofuran and water to give the acylated amino
~O acid of the formula
(XV)
Rlo
Il l 11
R~-~C~2) -C-N~-CH - C - OH
~5
The amino acid of formula XV is then coupled
to the amine of formula VI or VIII in the presence
of dicyclohexylcarbodiimide and l-hydroxybenzo-
triazole hydrate to give the desired compounds of
formula I.
The compounds of the ~ormula I wherein X is
o
Il
R~-(CH2)m-NH-C- and p is one can be prepared by
coupling an amino acid of the formula
1310792
HA375a
-15-
(XVI)
R5 O
Il l 11
6 (CH2)m-N~-c-NH-cH--C--OH
to the amine of formula VI in the presence of a
coupling agent such as dicyclohexylcarbodiimide
and l-hydroxybenzotriazole hydrate.
Similarly, the compounds of formula I
wherein X is
Q
Il . . .
R6- ( CH2 )m-NH-C-
and p is zero can be prepared by coupling an amino
acid of the formula
(XVII)
O R4 O
R6-(CH2)m-NH-C-NH--CH--C--OH
to an alcohol of formula II in the presence of a
coupling agent such as dicyclohexylcarbodiimide and
l-hydroxybenzotriazole hydrate.
The compounds of formula I wherein X is
O
CN_ C and p is one can be prepared by
coupling an amino acid of the formula
(XVIII)
O R5 O ~.
Il l 11
CN _ C _NH C~_ C _ OH
to the amine of formula VI in the presence of a
coupling agent such as dicyclohexylcarbodiimide
1 3 1 07q2
HA375a
-16-
and l-hydroxyben20triazole hydrate.
Similarly, the compounds of formula I
11
wherein X is C N - C- and p is zero can be
prepared by coupling an amino acid of the formula
~XIX)
0 R4 0
Il 1 11 .
CN_ C_ NH ~ CH - C -O~H
to the alcohol of formula II in the presence of a
coupling agent such as dicyclohexylcarbodiimide
and 1-hydroxybenzotriazole hydrate.
The amino acid intermediates of formulas
XVI, XVII, XVIII, and XIX can be prepared by treating
an amine R6-(CH2)m-NH2 with phosgene and N-methyl
~0 morpholine followed by reaction with an amino acid
methyl ester hydrochloride salt of the formula
(~)
R5 0
l 11
H2N-CH - C - 0 -CH3 ~Cl
or o the formula
(XXI)
R4 0
3a 1 11 ~.
H2N-CH - C -0 -CH3 ~ HCl
in the presence of N-methyl morpholine. Removal
of the methyl ester group by treatment with aqueous
3~ sodium hydroxide gives the desired intermediate.
1 3 1 07S2
HA375a
-17-
The products of formula I wherein
X is R5-(CH2)m CH -C-
I
R'
can be prepared by coupling the carboxylic acid
of the formula _~
(XXII)
o
Il
R5-(CH2)m CH- C -o~
(CH2)~,
I
R'6
to the amine of formula VI or VIII in the presence
of dicyclohexylcarbodiimide and l-hydroxybenzo-
triazole hydrate. Alternatively, the acid of
formula XXII can be converted to the acid chloride
and this acid chloride can then be coupled to the
amine of formula VI or VIII in the presence of
triethylamine and tetrahydrofuran or water and
sodium bicarbonate.
13~07q2
HA375a
-18-
The compounds of formula I wherein X is
O o
Il ~I 11
R6-(C~2)m- C - N - C~-C-
can be prepared by acylating proline with the acid
chloride of formula XII in the presence of base
such as sodium hydroxide, i.e., a pH of about 8~
and a solvent mixture of tetrahydrofuran and water
to giv~ ..
(XXIII)
O o
R6-(CH~)m C N - CH-C-OH
The acylated amino acid of formula XXIII is
then coupled to the amine of formula VIII or VI in
the presence of a coupling agent such as dicyclo-
hexylcarbodiimide and l-hydroxybenzotriazole
hydr~te.
~ he alcohol of formula II can be prepared by
treating the N-heterocyclic starting material of
the formula H-Rl with n-butyl lithium to yield the
lithium compound of the formula
~YIV)
Li-R
.
HA375al 3 1 0 7, 2
-19-
This lithium N-heterocyclic compound is then
reacted with the aldehyde o the formula
~YV)
R3 O
1 11
Prot-NH- CH -CH
wherein Prot is an amino protecting group such as
t-butoxycarbonyl. Removal of the t-butoxycarbonyl
group such as by treatment with hydrochloric acid
1~ givas the alcohol of formula II.
The aldehyde of formula XXV is prepared by .
treating the N-protected ~-amino acid ester of the
formula
(~XVI)
R3 O
l 11
Prot-NH-CH- C- O-alkyl
with lithium borohydride to give the alcohol of the
formula
~YVII)
R3
I
Prot -NH- CH - CH2 OH
Traatment of the alcohol of formula XXVII with
~5 pyridine-sulfur trioxide complex or with
periodinane reagent [see Dess et al., J. Org.
Che~., Vol~ 48, p. 5155-4156 (1983)] gives the
d~sired aldehyde.
In the above reactions, if any of R3, R4, R5
~nd Rlo are -(CH2)n-aryl wherein aryl is phenyl,
l-naphthyl, 2-naphthyl substituted with one or
1 3 ~ 0792
HA375a
-20-
more hydroxy or amino groups, -(CH2)n-heterocyclo
wherein heterocyclo is an imidazolyl, -(CH2)n~NH2,
( 2)n SH, (CH2)n~OH, -(CH2) -NH-C or
NH2
o
Il
-(CH2)n-C-OH then the hydroxyl, amino, imidazolyl,
mercaptan, carboxyl, or guanidinyl function should
be protected during the reaction. Suitable protect-
ing groups include benzyloxycarbonyl, t-butoxycarbonyl,
benzyl, benzhydryl, trityl, etc., and nitro in the
case of guanidinyl. The protecting group is removed
by hydrogenation, treatment with acid, or by other
known means following completion of the reaction.
The various peptide intermediates employed
in above procedures are known in the literature or
can be read~ly prepared by~known methods. See for
example, The Peptides, Volume 1, "Major Methods Of
Peptide Bond Formation", Academic Press (1979).
Preferred compounds of this invention axe
those of formula I wherein:
o
ll
X is R6 (C~2)m C
Rlo
R6 (CH2)m N~ C~ , R6-(CH2)m~~C~
1 3 1 ~7q2
-21- HA375a
o o
R6- ( CH2 )m-N}I-C- CN_C_ or
O
Il
6 ( H2)m ~CH C
(CIH2~m~
R6
R~ and R6' are independently selected from
straight or branched chain lower alkyl of up to 5
carbons, cycloalkyl of 4 to 6 carbons, phenyl,
l-naphthyl, and 2-naphthyl.
m and m' are independently selected from
zero, one and two.
C CN_ CN_
~o ~ /~ ~ ~
JW O N- S ~ N- HN N
H3C-N ~N-
~s
Especially preferred are the compounds
wherein X is O
3~
(H3C)3C-O-C-- ~ ~ ~ CH27~ CH-C- ,,
1 3 1 07~2
HA375
22 -
O o
l } c- , (H3C)3-C-CH2-C-
o
(H3C)3-C-NH-C-
o o o
(H3C)3-C-C- , C-N~-CH--C-
( CH2 ) 4
NH2
O O
CN_C_ O N-C-
\ -
f~
or H3C-N N-
~ 9 N ~ 9 ~3Rg
~R ~ N _ -R2
9 9
1 3 1 07~2
HA375a
-23-
Rg ~ ~ ~ Rg ,
Rg R2
~ Rg ~ Rg , especially
1 1
R2 2
~NI ~ ' ~ N
2 is CH2 CH2 ~ , hydrogen, straight
or branched chain lower alkyl of up to 5 carbons,
or -(CH2)n ~ wherein n is an integer from 1
- to 3.
1 3 1 0792
HA375a
-24~
Rg is hydrogen, straight or branched chain
lower alkyl of up to 5 carbons, or -~CH2)
S wherein n is an integer from 1 to 3.
R3 is straight or branched chain lower al~yl
of 3 to 5 carbons, -(CH2)n-cyclopentyl,
-(CH2)n-cyclohexyl, or -(CH2)n ~
wherein n is an integer from 1 to 3, especially -.
-CH2 ~ or -CH2CH(CH3)2
R4 is hydrogen, straight or branched chain
lower alkyl of up to 5 carbons, -(CH2)4-NII2 ,
r fJ 2 ~ ~N-cH2-o-cH2 ~ ,
-CX~ ~ ~ -CH2 ~ ~ _cu2 ~ OH ,
-C~ -C:~2~N , -Cn2~ '
1 3 ~ ~) 7
HA375a
-25-
-(C~2) ~ '` or -''^2
~ ~2
~2
especially 2 ~ ~J
R5 is strai~ht or branched chain lower alkyl
of up to 5 carbons,
-CH2 ~ , -(C:.2) ~ , -CH2 ~ - OCH3
-CH2-~-naphthyl), -CH2-(~-naphthyl), -CH2 ~ OH,
-CH2-cyclopentyl, -CH2-cyclohexyl~ C:' ~
-c-~2~ ,3
~ 2 ~ , especially benzyl.
y
1 31 07~2
HA375a
-26-
Rlo is (CH2)4 NH2
The compound of foxmula I form salts with a
variety of inorganic and organic acids. The non-
toxic pharmaceutically acceptable salts are pre-
ferred, although other salts are also useful in
isoiating or purifying the product. Such pharma-
ceutically acceptable salts include those formed
with hydrochloric acid, methanesulfonic acid,
sulfuric acid, acetic acid, maleic acid, etc. The
~alts are obtained by reacting the product with an~
equivalent amount of the acid in a medium in which
the salt precipitates.
The compounds o formula I contain asymmetric
centers when any or all of R3, R4, R5 and Rlo are other
than hydrogen and at the carbon to which the -OH
group i~ attached. Thus, the compounds of formula I
can axist in diastereoisomeric forms or in mixtures
thereof. The above described processes can utilize
racemates, enantiomers or diastereomers as starting
materials. When diastereomeric products are prepared,
they can be separated by conventional chromatographic
or fractional crystallization methods.
The compounds of formula I, and the pharma-
~5 ceutically acceptable salts thereof, are anti-
hypertensive agents. They inhibit the conversion
o~ angiotensinogen to angiotensin I and therefore,
are useful in reducing or relieving angiotensin
r~lated hypertension. The action of the enzyme
renin on angiotensinogen, a pseudoglobulin in
blood plasma, produces angiotensin I.
~ 3 ~ 0792
HA375a
-27-
Angiotensin I is converted by angiotensin
converting enzyme (ACE) to angiotensin II. The
latter is an active pressor substance which has
been implicated as the causative agent in several
S forms of hypertension in various mammalian
species, e.~., humans. The compounds of this
invention intervene in the angiotensinogen ~
(renin) ~ angiotensin I ~ tACE) ~ angiotensin II
sequence by inhibiting renin and reducing or
eliminating the formation of the pressor substance
angiotensin II. Thus by the administration of a _.
composition containing one (or a combination~ of
the compounds of this invention, angiotensin
dependent hypertension in a species of mammal
(e.g., humans) suffering therefrom is alleviated.
A single dose, or preferably two to four divided
d~ily doses, provided on a basis of about 100 to
1000 mg., preferably about 250 to 500 m~. per kg. of
body weight per day is appropriate to reduce blood
~0 pressure. ~he substance is preferably
administered orally, but parenteral routes such as
the subcutaneous, intramuscular, intraveneous or
intraperitoneal routes can also be employed.
The compounds of this invention can also be
~S formulated in combination with a diuretic for the
treatment of hypertension.
~ combination product comprising a compound
of this invention and a diuretic can be
administered in an efective amount which
comprises a total daily dosage of about 1000 to
~000 mg~, preferably about 3000 to 4000 mg. of a
I 3 1 07~2
HA375a
-28-
compound of this invention, and about 15 to
300 mg., preferably about 15 to 200 mg. of the
diuretic, to a mammalian species in need thereof.
Exemplary of the diuretics contemplated for use in
combination with a compound of this invention are
the thiazide diuretics, e.g., chlorothiazide,
hydrochlorothiazide, flumethiazide, hydroflu-
methiazide, bendroflumethiazide, methyclothiazide,
trichloromethiazide, polythiazide or benzthiazide
as well as ethacrynic acid, ticrynafen,
chlorthalidone, furosemide, mu~olimine, bumetanide,
triamterene, amiloride and spironolactone and salts
o~ such compounds.
The compounds of formula I can be formulated
for use in the reduction of blood pressure in
compositions such as tablets, capsules or elixirs
for oral administration or in sterile solutions or
suspensions for parenteral administration.
About 100 to 500 mg. of a compound of formula I is
compounded with physiologically acceptable
vehicle, carrier, excipient, binder, preservative,
stabilizer, flavor, etc., in a unit dosage form as
called for by accepted pharmaceutical practice.
The amount of active substance in these
~5 compositions or preparations is such that a
suitable dosage in the range indicated is obtained.
The following examples are illustrative of the
invention. All temperatures are given in degrees
centigrade.
HA37sa 7 3 ~ ~192
-29- -
Example
N -[N-[(1,1-Dimethylethoxy~carbonylL~L~ y~
alanyl]-N-[(S)-l-~hydroxy(lH-imidazol~2-yl~methyll-
3-methylbutYl~-L-histidinamide, ace~lc acid solvate
a) N-[(l,l-Dimethylethoxy)carbonyll-L-leucinal
Thionyl chloride (50.2 ml., 691 mmole) is
added dropwise over a period of 20 minutes to a
stirred (ice-cold) suspension of L-leucine
(72.05 g., 550 mmole) in absolute ethanol. After
the addition is completed, the ice-bath is removed
and the reaction mixture is stirred at room
temperature for one hour. It is then refluxed for
}ive hours on a steam bath. It is then
concentrated in vacuo and diluted with ether. The
-
~eparated crystals are filtered to give 99.9 g. of
L-leucine, ethyl ester, monohydrochloride; m.p.
~130) 135-137; [~]22 = +14.7 ~c = 2.3,
methanol),
A solution of L-leucine, ethyl ester, mono-
hydrochloride (46.85 g., 239.4 mmole) in 75%
ethanol (500 ml.) is added to a vigorously stirred
solution of sodium borohydride (35 g., 907 mmole)
in 75% ethanol (500 ml.) over a period of thirty
minutes. After the addition is completed, the
~5 reaction mixture is refluxed for 3 hours. Ethanol
is removed ln vacuo. The agueous solution is
extracted with ethyl acetate. The aqueous
solution is again extracted with ethyl acetate
after saturating with sodium chloride. The
combined ethyl acetate solution after washing with
saturated sodium chloride solution is evaporated to
HA375a 1 3 1 0 7 9 2
-30-
give 20.8 g. of crude (S~ 2-amino-4-methyl~l-pentanol.
Di-tert-butyl dicarbonate (38.73 g., 177.5
mmole) is added to a stirred (ice-bath) solution of
(S)-2-amino-4-methyl-1-pentanol (20.8 g.,
177.5 mmole) in tetrahydrofuran (340 ml.). After
stirring in an ice-bath for 15 minutes, the
reaction mixture is stirred at room temperature
for 3 hours. The tetrahydrofuran is removed ln
vacuo. This crude product (~0.53 g.) is combined
with 36.25 g. from a previous run and the entire
~lount is chromatographed over silica gel (800 g;)_~
using the solvent system ethyl acetate:hexane
(1:1) to yield 72.6 g. of (S)-2-[[(l,l-dimethyl-
ethoxy)carbonyl]amino]-4-methyl-1-pentanol as an
oil; [~]D2 = -26.2 (c = 1.5, methanol).
Trie~hylamine (63 ml., 450 mmole), dimethyl-
sulfoxide (63.9 ml., 900 mmole), and pyridine-
sulur trioxide complex (71.6 g., 450 mmole)
~re added to a stirred (room temperature) solution
of (S)-2-[[(1,1-dimethylethoxy)carbonyl]amino]-4
methyl-l-pentanol (32.59 g., 150 mmole) in
methylene chloride (750 ml.). After stirring the
reaction mixture for 15 minutes, it is evaporated
in vacuo at room temperature. Ice-water (450 ml.)
~5 is added to the residue which is then extracted
with ether. The e~her extract is successively
washed with 10% citric acid, water, saturated
sodium bicarbonate, and water. The ether extxact
on evaporat,on gives 19.73 g. of N-[(l,l-dimethyl-
ethoxy)carbonyl~-L-leucinal as an oil; ~]22 = _50O
(c = 4.6, methanol).
~ -31~ HA37sa 1 3 1 0 7 q 2
b) ~-~(S)-1-Amino-3-methylbutyll-1-[(phenyl-
methoxy)methyl~-lH-imidazole-2-methanol,
dihydrochloride
2.5 M n-Butyllithium solution in hexane
(4.6 ml., 11.5 mmole) is added to a solu~ion of
l-[(phenylmethoxy)methyl]-lH-imidazole (2.06 g.,
10.94 mmole; prepared as described by Brown et al.,
J.Chem.Soc. Perkin Trans. I, l9B2, p. 1553) in
tetrahydrofuran (35 ml.) at -70 under argon.
After 45 minutes at -70, an orange colored
solution results. A~ this point, a solution of --
N-[(1,1-dimethyleth~oy)carbonyl]-L-leucinal
(1.18 g., 5.47 mmole) in tetrahydrofuran (7 ml.)
is carefully added over a period of 2 - 3 minutes.
The reaction is kept at -70 for one hour, then at
0 for 15 minutes, and then is quenched by the
addition of saturated ammonium chloride (6 ml.).
The reaction mixture is diluted with ether and
rinsed with several portions of water and brine,
and dried over magnesium sulfate. Concentration
ln vacuo gives 3.13 g. of crude product. Flash
chromatography on silica gel (120 g. of Whatman
LPS-1) elutiny with petroleum ether-acetone t4:1)
gives 261 mg. of ~-[(S)-l-[[(l,l-dimethylethoxy)-
carbonyl]amino]-3-methylbutyl]-1-[(phenylmethoxy)
methyl]-lH-imidazole-2-methanol, fast moving
isomer; [a]22 = +5.8 (c - 0.5, chloroform),
564 mg. of a-[(S)-l-[[(1,1-dimethylethoxy)-
carbonyl]amino]-3-methylbutyl]-l~[(phenylmethoxy)-
methyl]-lH-imidazole-2-methanol, slow moving
isomer; [a]22 - +12.2 (c = 0.5, chloroform),
and 625 mg. of a mixture fraction for a total
1 3 1 0792
-32- HA375a
yield of 1.45 g. of product; TLC (silica gel;
petroleum ether:acetone, 3:1) Rf = 0.39, 0.13.
Anal. calc'd. for C22H3~N304:
C, 65.48; H, 8.24; N, 10.41
Found: C, 65.05; H, 8.20; N, 10.21
(fast moving isomer)
C, 65.26; H, 8.37; N, 10.29
(slow moving isomer)
A (1:1) isomeric mixture of the above fast
and slow moving isomers (493 mg., 1.22 mmole) is
dissolved in 10 ml. of ethyl acetate and cooled in_.
an ice-water bath under argon. The solution is
saturated with dry hydrochloric acid and stirred
cold for one hour, and then concentrated 1n vacuo
to give 420 mg. of crude product. Chromatography
on a 30 x 350 mm. HP-20 column gradient eluted
from 350 ml. of 9:1 to 1:9, 0.01 N aqueous hydro-
chloric acid:acetonitrile at 9 ml.~2 min./fraction
yields 402 mg. of a-[(S)-1-amino-3-methylbutyl-1-
[(phenylmethoxy)methyl]-lH-imidazole-2-methanol,
dihydrochloride; m.p. 98-117; [a]22 = _0 3O
(c = 1, methanol). TLC (silica gel; n-butanol:
pyridine:acetic acid:water, 4:1:1:1) Rf = 0.78.
Anal. calc'd for C17H25N3O2 2HC1 1.7 H2O:
~5 C, 50.17; H, 7.53; N, 10.33; Cl, 17.42
Found: C, 50.13; H, 7.54; N, 10.29; C1, 17.58.
c) N-~N-[(l,1-Dimethylethoxy)caxbonyl~-L-phenyl-
alanvll-l'-[(phenylmethoxy)methyl]-L-h-istldine
Thionyl chloride (27.2 ml., 375 mmole) is
added in drops to a stirred solution in an ice-
1 3 1 07q2
_33_ HA375a
bath of L-histidine (38.75 g., 2~0 mmole) in
methanol (500 ml.). After 15 minutes the ice-bath
is removed and the reaction mixture is stirred at
room temperature for one hour. After refluxing
for 48 hours, it is concentrated ln vacuo. The
separated crystals are filtered using methanol for
washings to 48.93 g. of L-histidine, methyl ester,
dihydrochloride. The methanolic solution on
dilution with ether affords an additional 10 g. of
product; m.p. 208 - 209; [~]2D2 = +10.1 (c = 1.8,
water).
Triethylamine (28 ml., 200 ml.) and di-tert-
butyl dicarbonate (48 g., 220 mmole) are added to a
suspension of L-histidine, methyl ester (24.~ g.,
100 mmole) in methanol (80 ml.). After 3.5 hours,
the mixture is filtered and the methanolic
solution is concentrated ln vacuo. The residue is
taken into chloroform and washed with 10~ citric
acid. The crude product on crystallization-from
isopropyl ether affords 23.1 g. of N,l'-bis~(l,1-
dimethylethoxy)carbonyl]-L-histidine, methyl ester;
m.p. (62) 88 - 95; [~]2D2 = +25 4 (c = 1 1
carbon tetrachloride).
Benzylchloromethyl ether (11.6 ml., 83.6 mmole)
is added to a solution of N,1'-bis[~1,1-dimethyl-
ethoxy)carbonyl]-L-histidine, methyl ester (24.7 g.,
66.9 mmole) in dry methylene chloride (156 ml.)
and the reaction mixture is stirred at room
temperature for 5 hours. After concsntrating ~n
vacuo and on dissolution in ethyl acetate 17.85 g.
of N-[~1,1-dimethylethoxy)carbonyl]-1'-[~phenyl-
methoxy)methyl]-L-histidine, methyl ester, mono
HA375a 1 3 1 07~2
-34-
hydrochloride crystallizes out; m.p. (148) 152 -
153; ~]D = -19.5~ (c = 1.8, methanol). This
methyl ester product is dissolved in hydrogen
chloride in acetic acid solution (60 ml., 1.5 N
and kept at room temperature for 15 minutes. It is
then evaporated in vacuo and the residue is
dissolved in hot isopropanol. After cooling, the
separated crystals are filtered to yield 7.08 g.
of l-[(phenylmethoxy)methyl]-L-histidine, methyl
ester, dihydrochloride; m.p. (170) 173 - 17~.
l-[(Phenylmethoxy)methyl]-L-histidine, methyl-
ester, dihydrochloride (1.79 g., 4.94 mmole),
l-hydroxybenzotriazole (0.756 g., 4.94 mmole), and
N-[(1,1-dimethylethoxy)carbonyl]-L phenylalanine
(1.31 g., ~.94 mmole) are dissolved in dimethyl-
formamide (16 ml.). While stirring the above
solution in an ice-bath, dicyclohexylcarbodiimide
(1.02 g., 4.94 mmole) and N,N-diisopropylethylamine
(1.72 ml., 10 mmole) are added. After 3 hours the
ice-bath is removed and the reaction mixture is
stirred at room temperature overnight. It is then
concentrated to dryness and the residue is tritura-
ted with ethyl acetate. The saparated urea is
filtered off. The ethyl acetate solution is
washed with saturated sodium bicarbonate and then
it is evaporated. The residue upo~ crystallization
from ethyl acetate gives 1.97 g. of N-[N-[(l,1-
dimethylethoxy)carbonyl]-L-phenylalanyl]~
~(phenylmethoxy)meth~l]-L-histidine, methyl ester;
m.p. (165) 166 - 168.
13107~2
HA375a
-35-
N-[N-[~l,1-Dimethylethoxy)carbonyl]-L-
phenylalanyl]-l'-[(phenylmethoxy)methyl]-L-
histidine, methyl ester (4.5 g., 8.4 mmole) is
dissolved in hot methanol (25 ml.). After cooling
to room temperature aqueous sodium hydroxide
solution t9.24 ml., lN) is added and the mixture
is stirred at room temperature for 3 hours. It is
then concentrated in vacuo and water (60 ml.) is
added to the residue. After cooling the aqueous
1~ ~olution in an ice-bath, it is acidified to pH 4~5
u~ing aqueous hydrochloric acid. It is then --
extracted with ethyl acetate to yield 3.95 g. of
crystalline N-[N-[~1,l-dimethylethoxy)carbonyl]-L-
phenylalanyl]~ [(phenylmethoxy)methyl]-L-
histidine; m.p. 193 - 194; [~]22 = -4.8
(c = 1.1, dimethylformamidè).
d) N -~N-[(l,l-Dimethylethoxy~carbonyl]-L-phenyl-
alanyll-N- r (s,-1- ~hydroxy[l-[(phenYlmethoxY)methvl~-
lH-imidazol-2-yllmethyl~-3-methylbutyl]-3'-[(phenvl-
~0 methoxy)methyl]-L-histidinamide
N-Methylmorpholine (154 mg., 1.52 mmole~ is
added to a mixture of N-[N-[(l,l-dimethylethoxy)-
carbonyl]-L-phenylalanyl]-l'-[(phenylmethoxy)methyl]-
L-hi~tidine (397 mg., 0.759 mmole), ~-[(S)-l-amino-3-
~S methylbutyl]-l-[(phenylmethoxy)methyl]~lH-imidazole-2-
methanol, dihydrochloride (309 mg., 0.759 mmole),
and l-hydroxybenæotriazole hydrate (116 mg.,
0.759 mmole) in dimethylformamide (5 ml.) cooled
in an ice-water bath under argon followed by the
30 addition of dicyclohexylcarbodiimide (157 mg.,
1 3 1 07q2
HA375a
-36-
0.759 mmole). The reaction mixture is kept cold
for 2 hours and then refigerated overnight. The
reaction mixture is then diluted with ethyl
acetate (30 ml.), chilled for 20 minutes, and then
filtered. The filtrate is further diluted with
ether and the organic solution is rinsed with two
portions of water (15 ml.), saturated sodium
bicarbonate solution (15 ml.), and brine, dried
over magnesium sulfate, and concentrated ln vacuo
to give 615 mg. of crude product. Two flash
chromatographies on silica gel (LPS-l) eluting .-
with chloroform:methanol:ammonia (25:2:0.05) gives
385 mg. of N2-~N~ dimethylethoxy)carbonyl]-L-
phenylalanyl]-N-[(S)-l-[hydroxy[l-[(phenylmethoxy)-
methyl]-lH-imidazol-2-yl]methyl]-3-methylbutyl]-
3'-[(phenylmethoxy)methyl]-L-histidinamide; m.p.
69-84; [~]D = -14.3 (c = 1, methanol). TLC
(silica gel; chloroform:methanol:ammonia,
25:2:0.05) Rf = 0.2, 0.26.
2 Anal- calc d- for C45H57N7O7 05 H2O
C, 66.15; H, 7.16; N, 12.00
Found: C, 66.20; H, 7.13; N, 11.73.
e) N2-[N-[(l,1-Dimethylethoxy)carbonyl]-L-phenYl-
alanyl]-N-[(S~ hydroxy(lH-imidazol-_-yl)methyl]-
3-methylbutyll-L-histidinamide, acetate
A solution of the product from part (d)
(337 mg., 0.413 mmole) in a mixture of methanol
(16 ml.), water (3.1 ml.), and lN aqueous
hydrochloric acid (0.908 ml.) is stirred under an
atmosphere of hydrogen (balloon) in the presence
o ?0% palladium hydroxide on carbon catalyst
1 3 1 37 9 ~
HA375a
-37
(80 mg.). After 17 hours, the reaction mixture is
filtered, concentrated ln vacuo, and flash chroma-
tographed on 36 g. of silica gel (LPS-l) eluting
with chloroform:methanol:water:acetic acid
(80:20:~.5:1). Pooliny of the product containing
fractions gives 187 mg. of N2-[N-[(l,l-dimethyl-
ethoxy)carbonyl]-L-phenylalanyl]-N-[(S)-1-
[hydroxy(lH-imidazol-2-yl)methyl]-3-methylbutyl]-
L-histidinamide, acetate; m.p. 90-118;
[~]D~ = -30.8~ (c = 0.5, methanol).
TLC`(silica gel; chloroform:methanol:water:acetic --
acid, 90:20:2.5:1) Rf = 0.14 and 0.17.
Anal. calc'd. for C29H41N7O5 2.4 C2 ~ 2
C, 54.2S; X, 7.36; N, 13.11
Found: C, 54.16; H, 7.01; N. 13.07.
Exam~le 2
N -~N-[(l,l-Dimethylethoxy)carbonyl]-L-~henyl-
alanyl]-N-~(S)-1-[(S)-hYdroxy(lH-imidazol-2-yl)-
methyll-3-me_h~vlbutyl~-L-histidinamide, acetic
?O acid solvate
"
a) NC-[N-L51,1-Dimethylethoxv)carbonyl]-L-~henyl-
alanyl~-N-[(S)-l-~(S~-hydroxy~ (phenylmethoxy)
m-thyl]-lH-imidazol-2-yllmethyll-3-methylbutyl~
3'- r (~henvl--m-ethoxy)methyl]-L-histidinamide
~5 A solution of ~-[(S)-1-[[(1,l-dimethylethoxy)-
c~rbonyl]amino]-3-methylbutyl]-1-[~phenylmethoxy)-
mathyl]-lH-imidazole-2-methanol, fast moving isomer,
from Example l(b), 320 mg., 0.79 mmole) in ethyl
acatate (7 ml.) is cooled in an ice-water bath and
saturated with gaseous hydrogen chloride. After
remaining in the bath for 15 minutes, the bath is
~ 3 1 37q2
HA375a
-38-
removed and the mixture is allowed to stand at
ambient temperature in a stoppered flask, for
65 minutes. The solution is concentrated 1n
vacuo to give 300 mg. of a solid white powder
residue of ~-[(S)-1-amino-3-methylbutyl]-1-
[(phenylmethoxy)methyl]-lH-imida~ole-2(S)-
methanol, dihydrochloride; m.p. 70-100~.
The above powder (270 mg., 0.66 mmole)
is dissolved in dry dimethylformamide (5 ml.).
1-H~droxybenzotriazole hydrate (100 mg.,
O.66 mmole), N-[N-[(1,l-dimethylethoxy)carbonyl]- '
L-phenylalanyl]~ [(phenylmethoxy)methyl]-L-
histidine ~342 mg., 0.66 mmole) and N-methyl-
morpholine (1~7 mg., 1.38 mmole) are added and the
mixture is cooled in an ice-water bath and treated
with dic~clohexylcarbodiimide (149 mg.,
0.66 mmole). The mixture is stirred, in a
stoppered flask, in the cold for 15 minutes and
then refrigerated for 18 hours. After diluting to
a volume of 35 ml. with ethyl acetate, the mixture
is filtered to remove insolubles and 35 ml. of
ether is added. After washing with water
(2 x 15 ml.), saturated sodium bicarbonate
solution ~15 ml.) and brine, the mixture is dried
~5 over magnesium sulfate and concentrated ln vacuo
to give 600 mg. of crude product as a viscous oil.
This residue is chromatographed on silica gel
(LPSl, 90 g.) eluting with chloroform:methanol:
con-entrated ammonium hydroxide (30:~:0.05) to
give 200 mg. of N2-[N-[(l,l-dimethylethoxy)-
carbonyl]-L-phenylalanyl]-N~[(S)-l-[(S)-hydroxy-
1 3 1 07S2
HA375a
-39-
[1-[(phenylmethoxy)methyl]-lH-imidazol-2-yl]-
methyl]-3-methylbutyl]~3'-[(phenylmethoxy)methyl]-
L-histidinamide as a glassy solid;
m.p. 70-100; [a]D = 27.5 (10 mg./ml. of
methanol). TLC (silica gel; chloroform:methanol:
concentrated ammonium hydroxide, 30:2:0.05)
Rf = 0.10.
Ana a 45 57 7 7
C, 65.86; H, 7.17; N, 11.95
Found: C, 65.86; H, 7.12; N, 11.76.
b) `N2-[N-[(1,1-DimethylethoxY)carbo~y~J-L-phenyl~
alanyl]-N-r(S)-l-[~S~-hydroxY(lH-imidazol-2-yl)methyll-
3-methylbutyll-L-histidinamide
To a solution of the product from part (a)
(194 mg., 0.236 mmole) dissolved in methanol
(8.9 ml.) is added water (1.7 ml.), lN a~ueous
hydrochloric acid (0.504 ml.) and 20% palladium
hydroxide on carbon catalyst (150 mg.~. The
mixture is stirred under an atmosphPre of hydrogen
(balloon~ for 18 hours. It is then filtered and
concentrated ln vacuo to give 153 mg. of crude
product. Flash chromatography (LPS-l silica gel,
25 g.) eluting with chloroform:methanol:water:
acetic acid (90:20:2.5:1) followed by lyophilli-
zation from 1% aqueous acetic acid of the pooledproduct containing fractions gives 124 mg. of
N2-[N-C(1,1-dimethylethoxy)carbonyl]-L-phenyl-
alanyl]-N-[(S)-l-[(S)-hydroxy(1~-imidazol-2-yl)-
methyl]-3-methylbutyl]-L-histidineamide, acetic
acid solvate; m.p. 105-110; [a]D ~ -30.1
1 3 1 07"2
HA375a
-40-
(~ - 0.5, methanol). TLC Isilica gel; chloroform:
methanol:water:acetic acid, 90:20:2.5:1) Rf = 0.11.
Anal calc'd. fox C~gH41N7O5~ 1.3 C2H4O~ 2
C, 55.23; H, 7.45; N, 14.27
Found: C, 55.19; H, 7.20; N, 14.29.
Example 3
N2-[N-[(1,l-Dimethylethoxy)carbonyl]-L-phen~
alanvl]-N-[(S)~1-[(R)-hydroxy~lH-imidazol-2-yl)-
methyll-3-methvlbutyl]-L-histidinamlde, acetic
acid solvate
a) N~-[N- L( 1,1-Dimethylethoxy)carbonyll-L-
phenylalanyll~N-~(S)-l-[(R)-hydroxy[l [(phenyl-
methoxy)-methyl]~lH-imidazol-2-yllmethyl]-3-methyl-
butyll-3'- E (phenylmethoxy)methyl]-L-histidinamide)
A solution of ~-[(S)-1-[[(1,l-dimethylethyl-
ethoxy)carbonyl]amino]~3-methylbutyl]-1-[(phenyl-
methoxy)methyl]-lH-imidazole-2-methanol,slow movin~
isomer, from Example l(b), (404 mg., 1 mmole) in
ethyl acetate (15 ml.) is cooled in an ice water
ba~h to 0 and saturated with gaseous hydrogen
chloride. After remaining in the bath for lS
minutes, the bath is removed and the mixture is
allowed to stand at ambient temperature for 45
minutes. Removal o the solvents ln vacuo gives
~5 396 mg. o~ solid a-[(S)-l-amino-3-methylbutyl]-1-
[(phenylmethoxy)methyl]-lH-imidazole-2(R)-methanol,
dihydrochloride.
To a mixture of this amine salt (0.81 mmole),
N-~N~tl,l-dimethylethoxy)carbonyl]-L-phenylalanvl]-
l'-[(phenylmethoxy?methyl]-L-histidine (423 mg.,
0.81 mmole), and 1-hydroxybenzotriazole hydrate
1 3 1 07q2
HA375a
-41-
(124 mg., 0.81 mmole~ in 6 ml. of dimethylformamide
cooled in an ice-water bath under argon is added
N-methylmorpholine (164 mg., 1.62 mmole) followed
by dicyclohexylcarbodiimide (167 mg., 0.81 mmole).
The reaction mixture is refrigerated overnight,
~len diluted with ethyl acetate and filtered. The
organic solution is rinsed with water, saturated
sodium bicarbonate solution, and brine, dried over
magnesium sulfate, and concentrated ln vacuo to
give 566 mg. of crude product. Flash chromato-
graphy (LPS-l silica gel, 60 g.) eluting with
chloroform:methanol:ammonia (30:2:0.05) gives
270 mg. of N2-[N-[(1,1-dimethylethoxy)carbonyl]-
L-phenylalanyl]-N-[(S)-l-[(~)-hydroxy~l-[(phenylmethoxy)-
methyl]-lH-imidazol-2-yl]methyl]-3-methylbutyl]-
3'-[(phenylmethoxy)methyl]-L-histidinamide;
m.p. 78-88; [~]D = -9.2 (c - 1,
methanol). TLC (silica gel; chloroform:methanol:
ammonia, 30:2:0.5) Rf = 0.10.
Anal. calc'd. for C45H57N707:
C, 66.89; H, 7.11; N, 12.14
Found: C, 66.69; H, 7.18; N, 12.17.
b) N2-[N-[(1,1-Dimethylethoxy)carbonylL-L-phenyl-
alanyl]-N-[(S)-1-[(R?~drox~y(1 -imidazol-2-yl)-
methyll-3-methYlbutyl]-L-histidinamide
To a olution of the product from part (a)
(255 mg., 0.316 mmole) dissolved in methanol
(11.7 ml.) is added water (2.2 ml.), lN agueous
hydrochloric acid (0.663 ml.) and 20% palladium
hydroxide on carbon catalyst (150 mg.). The
mixture is stirred under an atmosphere of hydrogen
1 3 1 0 7 9 L
HA375a
-42-
(balloon) for 18 hours. It is then filtered and
concentrated ln vacuo to give 208 mg. of crude
product. Flash chromatography (LPS-1 silica gel,
35 g.) eluting with chloroform:methanol:water:
acetic acid (90:20:2.5:1)followed by lyophillization
from 1% aqueous acetic acid of the pooled product
containing fractions gives 144 mg. of N2-[N-[(l,1-
dimethylethoxy)carbonyl]-L-phenylalanyl]-N-[(S)~1-
[(R)-hydroxy(lH-imidazol-2-yl)methyl]-3~methylbutyl]-L-
histidinamide, acetic acid solvate; _.
m.p. 194 - 196 (dec.); [~]D = 8.6
(c = 0.5, methanol). TLC (silica gel:chloroform:
methanol:water:acetic acid, 90:~0:2.5:1) Rf = 0.14.
Anal. calc~d~ for C29H41N7~5 1-3 C2H4~ 2
C, 55.22; H, 7.47; N, 14.36
Found: C, 55.15; H, 7.18; N, 14.31.
Example 4
N2-~N-L(l,1-Dimethylethoxy)carbonyl]-L-phenylalanyl]-
N-r(S)-1-~S?-hydroxy(lH_imidazol-2_ l)methyl~-2-~henyl-
ethYl~-L-histidlnamide, diacetate salt
a) N-[(1,1-Dim_thylethoxy)carbonyll-L-Phenylalaninal
To a solution of lithium borohydride (940 mg.
43.2 mmole) in dry tetrahydrofuran (90 ml.) cooled in
an ice-bath under nitrogen is added a solution
containing N-[(1,1-dimethylethoxy)carbonyl]-L
phenylalanine, N-hydroxysuccinimide ester (6.0 g.,
16.6 mmole) [prepared according to the procedure
of Anderson et al., JACS, Vol. 86, p.l839 (1964)] in
tetrahydrofuran (60 ml.). The addition is carried
out over a period of 5 minutes. After an
1 3 1 0792
HA375a
-43-
additional 20 minutes at 0, the reaction mixture
is poured into 1 1. of cold 10% potassium bisulfate.
The mixture is extracted with ethyl acetate (4 x
150 ml) and the combined organic extracts are
rinsed with saturated sodium bicarbonate, water,
and brine, dried over magnesium sulfate, and
concentrated ln vacuo to give 3.8 g. of crude
product. Flash chromatography (Merck 9385 silica
gel, ~50 g.) eluting with chloroform:methanol
(100:1, 50:1, and finally 25:1) gives 2.4 g. of
purified product. Recrystallization from ether-
hexane gives 1.9 g. of (S)-2-[[(1,1-dimethylethoxy)-
carbonyl]amino]-2-phenylmethyl-1-ethanol; m.p.
95-96; [~]D = -27.5 (c = 1, methanol). TLC
(silica gel; chloroform:methanol:acetic acid,
25:1:1) Rf = 0.54.
Anal. calc'd. for C14H21NO3
C, 66.90; H, 8.42; Ni 5.57
Found: C, 67.02; H, 8.49; ~, 5.23.
~0 Pyridine sulfur trioxide complex (3.04 g.,
19.1 mmole) is added to dry dimethylsulfoxide
~7 ml.) under argon and stirred at room
temperature for 15 minutes. This mixture is then
treated with a mixture of (S)-2-[[(1,1-dimethyl-
~5 ethoxy)carbonyl]amino]-~-phenylmethyl-l-ethanol
(1.~ g~, 4.77 mmole) and diisopropylethylamine
7 g., 19.1 mmole) in dry methylene chloride
~7 ml.), added rapidly along with the simultaneous
application of an ice-water cooling bath. The
ic~-bath is removed and after 10 minutes the
reaction mixture is poured onto a mixture of 50 ml.
1 :~1 07~2
EA375a
-44-
each of ice-water and ether. The aqueous portion
is further extracted with ether (2 x 50 ml.) and
the combined organic extracts are washed with 5%
potassium bisulfate, water, saturated sodium
bicarbonate, water, and brine, dried over
magnesium sulfate, and concentrated ln vacuo
to give 949 mg. of N-[(1,1-dimethylethoxy)
carbonyl]-L-phenylalaninal; m.p. 71-80;
[~]D = +37-7 (c = 1, methylene chloride).
TLC (silica gel; petroleum ether:acetone, 3:l)
Rf = 0.45.
b) N - [ ( S ? -1- ~ ~ 51, 1-Dimethylethoxy ? carbonyl~amino]-
2-phenylethyl-1-[(phenylmethoxy)methyll-lH-imidazole-
2-methanol
2.5 M n-Butyllithium solution in hexane
(2.8 ml., 6.99 mmole) is added to a solution of
l-[(phenylmethoxy)methyl]-lH-imidazole (1.28 g.,
6.82 mmole) in dry tetrahydrofuran (20 ml.) at
-70 under argon. After 45 minutes at -70, a
solution of N-[(1,1-dimethylethoxy)carbonyl]-L~
phenylalaninal (850 mg., 3.41 mmole) in tetra-
hydrofuran (4 ml.) is added over a period of
several minutes. After 2 hours at -70, the
reaction is warmed to 0 and then guenched by the
addition of saturated ammonium chloride (3 ml.).
The reaction mixture is then treated with ether
(lO0 ml.) and water (1.5 ml.). The organic
extract is rinsed with water (10 ml.) and brine,
dried over magnesium sulfate, and concentrated ln
30 vacuo to give 2.04 g. of crude product. Flash ~~
chromatography (LPS-1 silica gel, 80 g.) eluting
~ 31 0~7n~
HA375a
-45-
with petroleum ether:acetone (3:1) gives 234 mg.
oî ~-[~S)-l-[[(1,1-dimethylethoxy)carbonyl]amino]-
2-phenylethyl-1-[(phenylmethoxy)methyl~-lH-
imidazole-2-methanol (fast moving isomer),
152 mg. of a mixture fraction, and 294 mg. of a
slow moving isomer. TLC (silica gel; petroleum
ether:acetone, 2:1) Rf = 0.13, 0.08.
c) N~-[N- Ul,l-Dimethylethoxy)carbonYll-L-
phenylalanyll-N- r ( s ~ hydroxy[l~ henyl-
methoxy)methy,ll-lH-imidazol-2-yllmethyl]-2-
phenylethyll-3'~phenylmethoxy)methy~l]-L- --
histidinamide
An approximately (1:1) mixture of the fast
and slow moving isomer products from part (b)
(512 mg., 1.17 mmole) is dissolved in ethyl
acetate (20 ml.). The solution is cooled in an
ice-water bath and saturated ~ith gaseous hydrogen
chloride. The stoppered reaction is kept cold for
o~e hour, and then concentrated ln vacuo to
492 mg. of liyht tan colored a-[(S)-l-amino-2-
phenylethyl]-l-[(phenylmethoxy)methyl]-lH-
imidazole-2-methanol, dihydrochloride.
This amine salt (1.17 mmole) is heated with
l-hy~roxyben~otriazole hydrate (179 mg.,
~5 1.17 mmole), N-[N-[(l,l-dimethylethoxy)carbonyl]-
L-phenylalanyl]-l'-[(phenylmethoxy)methyl]-L-
histidine (611 mg., 1.17 mmole), and dimethyl-
~ormamide (9 ml.). The above mixture is cooled in
an ice-bath under argon and treated with dicyclo-
hexylcarbodiimide (237 mg., 1.17 mmole). The
reaction mixture is refrigerated overnight, then
1 3 1 0792
HA375a
-46-
diluted with ethyl acetate and filtered. The
filtrate is diluted with ether and xinsed with
water, saturated sodium bicarbonate, and brine,
dried over magnesi~ sulfate, and concentrated ln
S vacuo to give 882 mg. of crude product. Flash
chromatography (LPS-1 silica gel, 120 g.) eluting
with chloroform:methanol:ammonia (30:2:0.05)
yields 160 mg. of N2~[N-[(l,l-dimethylethoxy)-
carbonyl]-L-phenylalanyl]-N-[(S)-l-[hydroxy[1-
~(phenylmethoxy)methyl]-lH-imidazol-2-yl]methyl]-
~-phenylethyl]-3'-[(phenylmethoxy)methyl]-h-
histidinamide (fast moving isomer), 239 mg. of the
slow moving isomer, and about 60 mg. of a mixed
fraction. TLC (silica gel; chloroform:methanol:
ammonia, 30:2:0.05) Rf = 0.28, 0.26. [~]D = -25.4
(c = 1, methanol) for the fast moving isomer and
[~]D = -29.1 (c = 1, methanol) for the slow
moving isomer.
d) N2-[N-r(l,l-Dimethylethoxy)carbonyll-L-phenyl-
alanyll-N-[(S~ S)-hydroxy(lH-imidazo1-2-y~methyl]-
2-phenylethy~l_L-histidinamine, diacetate
salt
A mixture of the fast moving isomer from
part (c) (157 mg., 0.186 mmole), 20% palladium
~5 hydroxide on carbon catalyst (100 mg.), methanol
(6.9 ml~), water (1.3 ml.), and lN aqueous hydro-
chloric acid (0.391 ml.~ is stirred at room
temperature under hydrogen (balloon) for 25 hours.
The reaction mixture is then filtered and
30 concentrated in vacuo to give 139 mg. of crude ~-
-
product. Flash chromatography ILPS-silica gel,
13107~2
HA375a
-47-
21 g.) eluting with chloroform:methanol:water:
acetic acid (90:20:2.5:1) yields 107 mg. of
N2-[N-[~l,l-dimethylethoxy)carbonyl]-L-
phenylalanyl]-N-[(S)-l-[~S~hydroxy(lH-imidazol-2-yl)-
methyl]-2-phenylethyl]-L-histidinamide,
diacetate salt; m.p. 210-212 (d); [~]D = -21.4
(C = 0.5, methanol). TLC (silica gel; chloroform:
methanol:water:acetic acid, 90:20:2.5:1) Rf = 0.09.
Anal. calc~d. fox C32H39N7O5 2C~ 4 2 2
lOC, 57.06; H, 6.78; N, 12.94
Found: C, 57.01; H, 6.50; N, 12.94. ~-
Example 5
N2- rN- r (l,l-Dimethylethoxy)carbonvl]~L-phenyl-
alanyll-N-r(S)-l-[(R)-hydroxy(lH-imidazol 2-yl)-
methyl~-2-phenylethyl]-L-histidinamide
acetate salt ~1:1.5)
A mixture of N2-~N-[(1,l-dimethylethoxy)-
carbonyl]-L-phenylalanyl]-N-[(S)-l-[hydroxy[l-
[(phenylmethoxy)methyl]-lH-imidazol-2-yl]methyl]-
2-phenylethyl]-3'-~(phenylmethoxy)methyl]-L-
histidinamide (slow moving isomer) (236 mg.,
0.280 mmole) [prepared in Example 4(c)], 20%
palladium hydroxide on carbon catalyst (100 mg.),
methanol (10.4 ml.), water (2.0 ml.), and
lN aqueous hydrochloric acid (0.588 ml.) is
stirred at room temperature under hydrogen
(balloon) for 25 hours. It is then filtered and
concentrated ln vacuo to give 193 mg. of crude
product. Flash chromatography (LPS-l silica gel,
28 g.) eluting with chloroform:methanol:water:
acetic acid (90:20:2.5:1) yields 198 mg. of
1 3 1 07~2
HA375a
-48-
product. Lyophilli~atio~ from 1% agueous acetic
acid (polycarbonate filtered) yields 201 mg. of
N -[N~ dimethylethoxy)carbonyl]-L-phenyl~
alanyl]-N-[(S)-l~[(R)-hydroxy~ imidazol-2-yl)methyl]-
2-phenylethyl]-L-histidinamide, acetate
salt (1:1.5); m.p. 1g6 - 217; [~]D = ~30 7
(c = 0.5, methanol). TLC (silica gel; chloroform:
methanol:water:acetic acid, 90:20:2.5:1) Rf = 0.09.
~nal. calc'd. for C32H39N7s 1-5 C2H42 2
C, 57.47; ~, 6.81; N, 13.41
Found: C, 57.47; H, 6.47; N, 13.40.
Example 6
"
N -[N-[(l,l-Dimethylethoxy)carbonYll~L-phenyl-
alanyll-N-r(lS)-2-cyclohexy~ s-~-hy~ lH
lS imida~ol-2-vlmethylLethYl]-L-histidinamide,
monoacetate salt
a) (S~-2 LL~ _meth~ethoxy~arbonyl~amino
2-Phenylmethyl-1-ethanol
To a solution containing N-~(l,l-dimethyl-
ethoxy)carbonyl]-L-phenylalanine (lO g.,
37.7 mmole) in dimethylformamide (40 ml.) is added
solid sodium bicarbonate (4.75 g., 56.6 mmole~ and
iodomethane (16 g., 113 mmole). The mixture is
h~ated at ~0 under argon for 12 hours, ~he cooled
~S and the reaction mixture partitioned between water
(150 ml.) and ether (250 ml.). The organic layer
is rinsed with 2% aqueous sodium bicarbonat~
(2 x lO0 ml.), 2% aqueous sodium bisulfite
(lO0 ml.),~water (2 x 100 ml.)~ and brine, dried
over magnesium sulfate, and concentrated ln vacuo
to give 10.5 g. of N-[(1,l-dimethylethoxy)carbonyl]-
1 ~` 1 979
HA375a
-49-
L-phenylalanine, methyl ester as an oil;
[~]D = +47-7 (c = 1, methylene chloride3
TLC (silica geli petroleum ether:acetone,
6:1) Rf = 0.41.
Anal. calc'd. for C15H21NO4:
C, 64.49; H, 7.58; N, 5.01
Found: C, 64.56; H, 7.60; N, 5.29.
To a solution containing N~[(l,1-
dimethylethoxy)carbonyl]-L-phenylalanine, methyl
ester (10 g., 35.8 mmole) dissolved in a mixture of
tetrahydrofuran (190 ml.) and absolute ethanol
(190 ml.) is added lithium chloride (6.09 g.,
143.2 mmole). The resulting homogeneous solution
is treated with sodium borohydride (5.42 g.,
143.2 mmole) and the reaction is stirred at room
temperature under argon for 24 hours. The
reaction mixture is next filtered using ether
(a'oout 700 ml.) to rinse the filter cake. The
resulting filtrate is rinsed with water ( 3 x
200 ml.~ and brine (200 ml.), dried over magnesium
sulfate, and concentrated 1n vacuo to give 9 g. of
crude product. Recrystallization from
ether/hexane gives 7.59 g. of (S)-2-[[(1,1-
dimethylethoxy)carbonyl]amino]-2-phenylmethyl-1-
ethanol; m.p. 94 - 96; [a]D ~ -27-2 (c = 1,
methanol). TLC(silica gel; petroleum
ether:acetone, 3:1) Rf = 0.39.
Anal. calc'd. for C14H21NO3:
C, 66.90; H, 8.42; N, 5.57
Found: C, 66.80; H, 8.57; N, 5.38.
- 1 3 ~ 07q2
HA375a
-50-
b) [(S)-2-Cyclohexyl-1-(hydroxymethyl)ethyl
carbamic acid, 1,1-dimethylethyl ester
A solution of (S)-2-[[(1,1-dimethylethoxy)-
carbonyl~amino]-2-phenylmethyl-1-ethanol (7 g.,
27.8 mmole) in methanol (70 ml.) is hydrogenated
at 55 psi on a Parr Shaker using 5% rhodium on
alumina (500 mg.) as catalyst. After 17 hours,
the reaction mixture is filtered and concentrated
in vacuo to yield 7.36 g. of [(S)-2-cyclohexyl-1-
(hydrox~ethyl)ethyl]carbamic acid, 1,1-dimethyl-
ethyl ester as an oil; [~]D = -23.3 (c = 1,
methylene chloride). TLC (silica gel; petroleum
ether:acetone, 3:1) Rf = 0.5.
Anal. calc'd. for Ci4H27NO3:
C, 65.33; H, 10.57; N, 5.44
Found: C, 64.94; H, 10.55; N, 5.23.
c) (S)-(2-Cyclohexyl-1- I ormylethyl~carbamic acid,
l,l-dimethyleth~l estex
A solution of [(S)-2-cyclohexyl-1-(hydroxy-
methyl)ethyl]carbamic acid, l,l-dimethylethyl
ester ~4.6 g., 17.9 mmole) in methylene chloride
(40 ml.) is added to a mixture of Dess-Martin
periodinane reagent (8 g., 19 mmole) [prepared
according to Dess et al., J. Org, Chem., Vol. 48,
p. 4155 (1983)] and t-butanol (1.5 g., 19 mmole)
in methylene chloride ~70 ml.) which had been
stirred at room temperature before the addition.
A slight exotherm (to 32) results. After 30
minutes, the reaction mixture is quenched in ether
(800 ml.), resulting in the separation of a white
solid. A mixture of sodium thiosulfate pentahydrate
7 ~ ,~
HA375a
-51-
(31.3 g., 126 mmole) in saturated sodium bicar-
bonate solution (200 ml.) is added, with stirring.
The resulting two-phase mixture is separated and
the oxganic phase is washed with water, saturated
sodium bicarbonate (2 x 100 ml.), water, and
brine, dried over magnesium sulfate, and
concentrated ln vacuo to give 3.8 g. of (S)-(2-
cyclohexyl-l-formylethyl)carbamic acid, 1,1
~imethylethyl ester as a colorless oil.
1~ d) ~(lS)-l-(Cyclohexylmethyl)-2-hydroxY-2-
~1-[(phenvlmethoxy)methylJ-lH-imidazol-2-yl]-
ethyl~carbamic acid, l,1-dimethyleth~l ester
2.5 M n-Butyllithium solution in hexane
(12 ml./ 31 mmole~ is added to a solution of
l-[(phenylmethoxy)methyl]-lH-imidazole ~5.3 g.,
28 mmole) in tetrahydrofuran (90 ml.) at -70
under argon. After stirring for 15 minutes,
(S)-(2-cyclohexyl-1-formylethyl)carbamic acid,
l,l-dimethylethyl ester (3.6 g., 14 mmole) in
0 tetrahydrofuran (36 ml.) is added dropwise over a
period of 5 minutes at a reaction temperature of
-65 to -70. After 2 hours at ~70, the bath is
warmed to 0 and saturated ammonium chloride
(25 ml.) is added followed by ether (300 ml.) and
water (25 ml.). The organic phase is washed with
water (2 x 50 m.) and brine, dried over magnesium
sulfate, and concentrated ln vacuo. The resulting
crude product (8.4 g.) is flash chromatographed
~LPS-l silica gel) eluting with acetone:petroleum
ether ~1:4) to give 580 mg. of [(lS)-1-(cyclo- ~
hexylmethyl)-2-hydroxy-2-[1 [(phenylmethoxy)methyl]-
~ 3 ~ O~q2
HA375a
-52-
lH-imidazol-2-yl]ethyl]carbamic acid, 1,1-
dimethylethyl ester (fast moving isomer), 370 mg.
of a mixed fraction, and 2 g. of a slow moving
isomer. TLC (silica gel; acetone:petroleum ether
1:4) Rf = 0.15, 0.10.
Fast moving isomer; [~]D = -21.5
(13 mg./ml., methanol).
Anal.calc'd. for C25H37N3O4 2
C, 66.61; H, 8.45; N, 9.32
Found: C, 66.55; H, 8.39; N, 9.00.
Slower moving isomer; [~]D = +9.1 -'
(14 mg./ml., methanol).
Anal- calc'd. for C25H37N304 022 H2O
C, 67.08; H, 8.43; N, 9.39
Found: C, 67.08; ~, 8.35; N, 9.01.
e~ N2-~N-[(l~l-Dimethylethoxy)carbonyl]-L-
henylalanyll-N-~(S~-l-r(S)-hydroxy11-[(phenyl-
methoxy)methyl~-lH-imidazol-2-~l]methYl]-?-
cyclohexylethyll~3'-[(phenylmethox~methyl~-L-
histidinamide
A solution of the fast moving isomer from
part (d) (430 mg., 0.97 mmole) in ethyl acetate
(8 ml.) is cooled in an ice-water bath and
saturated with gaseous hydrogen chloride. After
remaining in the cold for 15 minutes, the mixture
is kept in a stoppered flask at ambient
temperature for 45 minutes. The mixture is
concentrated ln vacuo to give 384 mg. of the amine
dihydrochloride salt.
This amine salt (306 mg., 0.8 mmole) is
dissolved in dry dimethyformamide (6 ml.) and
13tO7q2
HA375a
-53-
N-[N-[(l,l-dimethylethoxy)carbonyl]-L-phenyl-
alanyl]-1'-[(phenylmethoxy)methyl~-L-histidine
(442 mg., 0.8 mmole) is added followed by
l-hydroxybenzotria201e hydrate (122 mg.,
0.8 mmole) and N-methylmorpholine (160 mg.,
1.6 mmole). After cooling in an ice-water bath,
dicyclohexylcarbodiimide is added. The mixture is
~tirred in the cold for 15 minutes and then
refrigerated overnight in a stoppered flask. The
solids that separate ~rom the solution are
recovered by filtration after the mixture is
diluted to a volume of 45 ml. with ether. The
filtrate is washed with water, sa~ura~ed sodium
bicarbonate solution, and brine, dried over
magnesium sulfate, and concentrated in vacuo to
give a viscous oil residue (587 mg.). This
residue is preabsorbed on silica gel (Baker's,
3 g.~ and flash chromatographed (LPS-1 silica gel,
90 g.) eluting with chloroform:methanol:concentrated
~0 ammonium hydroxide ~30:2:0.05) to give reco~ery of
the product in two fractions (333 mg. of impure and
70 mg. of pure product). The impure fraction is
rechromatographed as above to give a recovery o~
206 mg. This is combined with the 70 mg. fraction
~S and chromatographed again according to the above
procedure to give 255 mg. of N2-[N-[(l,1-dimethyl-
e~hoxy)carbonyl]-L-phenylalanyl]-N-[(lS)-1-[~S)-
hydroxy-[l-[(phenylmethoxy)methyl]-lH-imidazol-2-
yl]methyl]-2-cyclohexylethyl]-3'-[~phenylmethoxy)-
mathyl~-L-histidinamide as a glassy solid;
m.p. 74-77 ~gradual melt); [a]D = -24.3
1 3 ~ 079~
~3753
. SD~ - .
~c = 1, methanol3. TLC (silica g~l: chloroo~s
math3nol:conc. ammo~iu~ hydroxid~, 30:2 0.0
R~ = û.17.
Anal . calc ' d- for C4~3E3t61N7~7 1E~2
C, 66,57; ~, 7.33; ~ 11.32
Found: C, 66.55; E~, 7.26; N, 11.37.
f) ~__
A mixt~re~ of th~ produc~ ~o~
part ~) (24~ mg., 0.~8 m~ola) ~ 20% ~alladl~u~
h~dro~ida oQ car~o~ ~ataly~ 185 ~g.) in m~ o~
~i ml.~ ~lu5 wat~r (0.~1 ~O) ~nd l~t h~lkos:l~lor~c
acid (0.62 ml., 0.62 ~oles) i4 8tl~0~ at roo~a
tllperatur~ in a~ at~o~ o~ hydrog~~ lloo~
for 60 hour~ addition~l 100 n~. s:~ ca~ly~t
and 4 ml. o~ m*thanol ar~ ad~3d a~
stir~ed in a~ atmosph~ of hy~go~ ~or 20
hour~. After filterin51 ~ough~ite~, tl~
filt~at2 i~ eonc~era~d i_ ~ to qiv~$ 200 mg,
o~ ~ gl~s~y solid seEI~du~. Thi~ ros~ldu~ i~ th~
fla~h c~ro~atoslraph~a ~I,PS~ ca ~ 0 sr~)
~lutiny wit~ c3~10ro~on~:mothanol :wat~s a~otlc: ~id
~9Os20:2.Ssl) to gi~ 120 Dl~. o~ h ~ r no~,id
25 ~ ak i~ lyol?hilliz~d froDI~ 3X alC~ cia to givo
_120 mg. o~ an amorphous whits Jol~d. Thi~ ~olid
i~ ~o~hro~atograph~c~ (IPS-l ~ilic~ gol, 16 ~. )
eluting with chloroform:m~ ol:w~t~r:ac~
acid ~90:20:205:1) to give~ 69.4 m~. of N2 tN~
30 dimcthyl~l:ho~y~carbo~yl~-L-phsnylalanyl]-N-t~15) 2-
* Trademark
~ .
~ ' ,.
1 3 1 079~
HA375a
-55-
cyclohexyl-1-[(S)-hydroxy-lH-imidazol-2-ylmethyl]-
ethyl] L-histidinamide, monoacetate
salt; m.p. 162 (gradual melt, shrinks at 110);
[~]D = -35.6 (5 mg./ml., methanol). TLC (silica
gel; n-butanol:pyridine.acetic acid:water,
4:1:1:1) Rf = 0.61.
Anal. calc'd- for c32H45N7o5 1 C2 4 2
C, 57.07; H, 7.65; N, 13.70
Found: C, 57.05; H, 7.40; N, 13.72.
10Example 7
N2 [N-~(l,l-Dimethylethoxx~ y~
alanyll-N-~lS)-2-cy~clohexyl-l-[(R)-hydroxy-lH
imidazol-2-ylmethyllLethyll-L-histidinamide~
monoacetate salt
a) N2-rN-r(l,l-DimethYlethoxy)carbonyll-L-
phenylalanyl]-N-[(lS)-l- L t R)-hydroxv~ phenYl-
methoxv)methyl~-lH-imidazol-2-yl]meth~yl]-2-
cyclohexylethyll-3~ henylmetho-xy)me-thyl]-L
histidinamide
A solution of [(lS)-1-(cyclohexylmethyl)-
2-hydroxy-2-[1-[(phenylmethoxy)methyl]-lH-
imidazol-2-yl]ethyl]carbamic acid, 1,1-
dimethylethyl ester (slow moving isomer)
(467 mg., 1,05 mmole) [prepared in Example 6 (d)]
in ethyl acetate (25 ml.) is cooled in an ice-
water bath under argon and saturated with gaseous
hydrogen chloride. The stoppered reaction is kept
cold for one hour and then concentrated ln vacuo
to give 467 mg. of the amine dihydrochloride salt.
This amine salt (374 mg., 0.84 mmole) is ~~
dissolved in dimethylformamide (6 ml.) along with
~310:7~
HA375a
-56-
N-[N-[(l,l-dimethylethoxy)carbonyl]-L-phenyl
alanyl]-l'-[(phenylmethoxy)methyl]-L-histidine
(439 mg., 0.84 mmole) and l-hydroxybenzotriazole
hydrate (128 mg., 0.84 mmole). The mixture is
cooled under argon in an ice-water bath and
treated with N-methylmorpholine (170 mg.,
1.68 mmole) followed by dicyclohexylcarbodiimide
(173 mg., 0.84 mmole). The stoppered reactlon
mixture is refrigerated o~ernight, then filtered
and extracted with ethyl acetate. The organic ~.
solution is rinsed with water, saturated sodium
bicarbonate, water, and brine, dried over maynesium
sulfate, and concentrated ln vacuo to 730 mg. of
crude product. Flash chromatography (LPS-l silica
gel, 70 g.) eluting with chloroform:methanol:
concentrated ammonia (30:2:0.05) gives 271 mg.
of N2-[N-[(1,1-dimethylethoxy)carbonyl]-L-phen~l-
alanyl]-N-[(lS)-l-[(R)-hydroxy[l-[(phenylmethoxy)-
methyl]-lH-imida~ol-2-yl]methyl]-2-cyclohexyl-
ethyl]-3'-[(phenylmethoxy)methyl]-L-histidinamide;
[~]D = -8.7 ~c = 1, methanol).
TLC ~silica gel; chloroform:methanol:conc. ammonia,
30:2:0.05) R~ = 0.14.
Anal. calc'd. for C48H61N7O7:
~5 ~, 67.98; H, 7.25; N, 11.56
Found C, 67.80; H, 7.27; N, 11.44.
b) N -[N-~(l,l-Dimethylethoxy ? carbonyl]-L-phenyl~
alanyl~-N-[(lS ? -2-cyclohexyl-1-[(R~-hydroxy-lH-
imidazol-2-ylmethylle-hyl]-L-histidinamide
3~ monoacetate salt ~`
.
The product from part (a) (265 mg.,
0.312 mmole) is dissolved in methanol (11.6 ml.)
1 3 1 07q2
_57_ HA375a
to which is added water (2.2 ml.) followed by
lN aqueous hydrochloric acid ~0.66 ml.) and 20%
palladium hydroxide on carbon catalyst (100 mg.).
The mixture is stirred under a hydrogen atmosphere
(balloon) for 17 hours, then filtered and
concentrated in vacuo to give 250 mg. of crude
product. Flash chromatography (LPS-l silica gel,
35 g.) eluting with chloxoform:methanol:water (tap
distilled):acetic acid (90:20:2.5:1) followed by
lyophillization of the product containiny
fractions gives 186 mg. of material. Rechroma- --
tography using double distilled water (LPS-l
silica gel, 20 g.) eluting with the 90:20:2.5:1
solvent system gives 95 mg. of N2-[N-[(l,1-
dimethylethoxy)carbonyl]-L-phenylalanyl]-N-
[(lS)-2-cyclohexyl-1-[(R)-hydroxy-lH-imidazol-2-
ylmethyl]ethyl]-L-histidinamide,
monoacetate sal~; m.p. 193 - 197 (d 200);
[~]D = -19 (c = 0.5, methanol). TLC (silica
gel; chloroform:methanol:water:acetic acid,
90:20:2.5 1) Rf = 0.07.
Anal- calc'd- for c3~H45N7O5 0-8 C2 4 2
C, 59.26; H, 7.55; N, 14.40
Found: C, 59.24; H, 7.49; N, 14.37.
25Example 8
N-[(lS?-2-C clohex~l-l-[(R)-hydroxY(lH-imidazol-2-yl)~
methyl]ethy~ N2~[N~ l-dimethylethoxy)carbonyll-L
phenylalanyll-L-leucinamide~ acetate salt
a) N-[N-r(l,l-Dimethvlethoxy~ rbonyl~-L-
30 ~ phenylalanyl]-L-leucine~ methyl ester
A solution of diisopropylethylamine (8.7 ml.,
50 mmole) in tetrahydrofuran (50 ml.) is added
dropwise to a mixture of N-[(l,1-dimethylethoxy)-
carbonyl~-L-phenylalanine (13.265 g., 50 mmole),
L-lPucine, methyl ester (9.085 g., 50 mmole) and
1 3 1 07~2
HA375a
-58-
l-hydroxybenzo~riazole hydxate (7.65 g., 50 mmole)
in tetrahydrofuran (lO0 ml.) at 0. This is
followed by the addition of dicyclohexylcarbo-
diimide (10.315 g., 50 mmole). The reaction is
stirred at 0 for 2 hours and left stirring
overnight at room temperature. The next day, the
precipitated dicyclohexyl urea is filtered off,
the solvents are stripped down and the residue is
diluted with ethyl acetate (200 ml.). The organic
solution is washed sequentially with saturated
aqueous sodium bicarbonate solution (2 x 100 ml.)
and saturated aqueous sodium chloride
(2 x lO0 ml.), dried over sodium sulfate,
filtered, and concentrated to give crude product.
~5 Crystallization from ethyl ether gives 7.05 g. of
pure product. Concentration of the mother liquor
solutions gives 4.57 g. of crystalline product. An
additional I.35 g. of product is obtained by
chromatographic purification of the crude product
obtained from the left-over mother liquors (40 g.
silica gel, eIuting with 4:1 hexane:ethyl acetate).
Thus, a total of 12.96 g. of N- [N- [(1,1-dimethyl-
ethoxy)carbonyl]-L-phenylalanyl]-L~leucine, methyl
ester is obtained; m.p. 104 - 105; [~]D = -17.5
~5 (c = 1.2, methanol). TLC (silica gel; hexane:
ethyl acetate, 1:1) Rf - 0.57.
Anal. calc'd. for C21H32N2O5:
C, 64.30; H, 8.15; N, 7.14
Found: C, 64.12; H, 8.16; N, 7.02.
b) N-~LlS)-2-Cyclohexyl-l-[(R)-hydroxy[l-[(pheny
methoxy)methyl]-lH-imidazo
N2-[N-~(l,l-dimethylethoxy)carbonyl]-L-~henyl-
alanyl]-L-leucinamide
The methyl ester product from part (a)
(1.176 g., 3 mmole) in tetrahydrofuran (12 ml.)
I 3 1 0792
HA375a
-59-
is treated with aqueous lN sodium hydroxide
(3.3 ml., 3.3 mmole). After two hours at room
temperature, the mixture is refluxed for two
hours The solvents are stripped down and the
residue is taken up in water, acidified, and
extrac~ed with ethyl aceiate. The organic layer
is dried over sodium sulfate and concentrated to
give a residue which is resubjected to hydrolysis
with aqueous lN sodium hydroxida (1.5 ml.,
1.5 mmole) in methanol (12 ml.). After three
hours at room temperature, the solvents are
stripped down. The residue is taken up in water
~100 ml.), extracted with ethyl acetate
t2 x ~5 ml.), and the organic layer is discarded.
The aqueous layer is carefully acidified (pH 3.9),
extracted with ethyl acetate (3 x 25 ml.), dried
over sodium sulfate and concentrated to give
~53 mg. of N-[N-(1,1-dimethylethoxy)carbonyl]-L-
phenylalanyl]-L-leucine.
~0 To a mixture of this acid (753 mg.,
2.0 mmole), [(lS)-1-(cyclohexylmethyl)-2-hydroxy-2-
[l-[~phenylmethoxy)methyl]-lH-imidazol-2-yl]ethyl]-
carbamic acid, l,l-dimethylethyl ester, slow isomer
~rom Example 6(d) (887 mg., 2.0 mmole), and
~S l-hydroxyben~otriazole hydrate (306 mg., 2.0 mmole)
in ~etrahydrofuran (8 ml.) at 0 is added diiso-
propylethylamine (731 ~1., 4.2 mmole) followed by
dicyclohexylcarbodiimide (416 mg., 2.0 mmole).
T~e reaction mixture is stirred at 0 for about
2 hours and then kept overnight in the cold room
(about 5). The next day, the precipitated dicyclo~
hexyl urea is filtered off and the residue is
redissolved in ethyl acetate, washed with
saturated aqueous sodium bicarbonate solution
(2 x 30 ml.) and saturated aqueous sodium chloride
I 3 1 0792
HA375a
-60-
(2 x 30 ml.), dried over sodium sulfate, filtered,
and c~ncentrat~d to give 1.339 g. of a crude
residue. Repeated flash chromatographies (silica
gel, eluting with 9:1:0.1 and 15:1:0.05
chloroform:methanol:acetic acid) gives 1.021 g. of
N-[(lS)-2-cyclohexyl 1-[(R)-hydroxy[l-[(phenylmethoxy)-
methyl]-lH-imidazol-2-yl~methyl]ethyl]-N2-[N-[(1,1-
dimethylethoxy)carbonyl]-L-phenylalanyl]-L-leucinamide.
TLC (silica gel; chloroform:methanol:acetic acid,
9:1:0.1) Rf = 0.5.
c) N-~(lS) c-~-lo>e~y~ l IlR)-hydroxy~lH-imidazol-
2-yl)methyl]ethyl]-N2-[N-~(l,l-dimethylet oxy)car-
bonyl]-L-phenvlalanyll-L-leucinamide, acetate salt
20% Palladium hydroxide on carbon catalyst
(100 mg.) is weighed in a side arm flask and flushed
with hydrogen using a balloon. Ths L=leucinamide
product from part (b) (295 mg., 0.42 mmole) is
added via a syringe as a solution in methanol
(5 ml.). This is followed by sequential addition
of mathanol (8.5 ml.~, water (3.0 ml.), and
aqueous 10% hydrochloric acid (0.307 ml.). The
flask is then carefully flushed several times with
hydrogen and stirred overnight at room temperature.
The contents of the flask are filtered and the
filtrate is concentrated. The resulting residue
is flash chromatographed (30 g. of silica gel;
elutins with 90:10:1:0.1 chloroform:methanol:
water:acetic acid) to give 155.3 mg. of N-[(lS)-
2-cyclohexyl-1-[(R)-hydroxy(lH-imidazol-2-yl)methyl]-
ethyl]-N2-[N-[(1,1-dimethyle~hoxy)carbonyl]-L-
phenylalanyl~-L-leucinamide, acetate
salt; m.p. 101-109 (gradual melting); [a]D = -25.7
(c = 1.06, methanol). TLC (silica gel; chloroform:
~ 3 t 07q2
HA375a
-61-
methanol:water:acetic acid, 90:20:2.5:0.1) Rf = 0.21.
Anal- calc'd- for C32H~9N55 0 9 C2 4 2 2
C, 6~.33; H, 8.37; N, 10.75
Found: C, 62.38; H, 8.17; N, 10.75.
Example 9
N-[(lS)-1-(Cyclohexylmethyl)-2(R)~hydroxy~2-(lH-
imidazol-2-yl)ethyl]-N2-[N-[(l,1-dimethylethoxy)-
carbonyl~-L-phenylalanyl]glycinamide, acetate salt
a) N-[N-[(l,1-Dimethylethoxy)carbonyl]-L-~henyl-
alanyl~glycine~ ethyl ester
To a mixture of N-[(1,1-dimethylethoxy)- -
carhonyl]-L-phenylalanine (13.265 g., 50 mmole),
glycine, ethyl ester, monohydrochloride (6.98 g.,
50 mmole), and l-hydroxybenzotriazole hydrate
~7.65 g., 50 mmole) in tetrahydrofuran (100 ml.)
at 0 is added dicyclohexylcarbodiimide (10.315 g.,
50 mmole~ as a solution in tetrahydrofuran (25 ml.).
This is followed by the addition of a solution of
diisopropylethylamine (8.7 ml., 50 ~mole) in tetra-
hydrofuran (25 ml.). The rea`ction is stirred at 0
for 2 hours and then stirred ovexnight at room
temperature. The next day, the precipitated
dicyclohexyl urea is filtered off and the solvents
~5 stripped down. The residue is diluted with ethyl
acetate (200 ml.) and the resulting organic
301ution is washed sequentially with saturated
a~ueous sodium bica.rbonate solution (2 x 100 ml.)
and saturated aqueous sodium chloride (100 ml.),
dried over sodium sulfate, filtered, and concen-
trated. The resulting crude material is crystall-
ized from ethyl ether to give 9.45 g. of pure
product. Concentration of the mother li~uor
1 3 1 07"2
HA375a
-62-
solution to half its original volume gives an
additional 3.7 g. of N-[N-[(1,1-dimethylethoXy)-
carbonyl]-L-phenylalanyl]glycine, ethyl ester;
m.p. 90~9~; [~]D = -9.1 (c = 1.41, methanol).
Anal. calc'd. for C18H26N2O5:
c, 61.73; ~, 7.42; N, 7.99
Found: C, 61.64; H, 7.46; N, 8.07.
b) N-[(lS)-l-(Cyclohexylmethyl)-2(R)-hydroxy-
~-[1-[(Phenylmethoxy)methyl]-lH-imidazol-2
ethyll-N2-[N-[(l,l-dimeth~lethoxy~carbonyl]-L~
~henylalanyl]glycinamide
A solution of the ethyl ester product from
part (a) (1.05 g., 3.0 mmole) in tetrahydrofuran
(12 ml.) is`treated with agueous lN sodium
hydroxide (3.1 ml., 3.1 mmole). ~ydrolysis is
found to be complete after 2 hours at room tempera-
ture. The solvents are stripped down and the
residue is taken up in saturated agueous sodium
bicarbonate solution (15 ml.) and extracted with
ether (30 ml.). Some sodium salt precipitates at
this stage and is filtered and separated from the
biphasic layer. The ethereal layer is discarded.
The aqueous layer is carefully acidified (pH 5.0
and finally 2.5) and reextracted with ethyl acetate
(3 x 20 ml.). The precipitated sodium salt is
taken up in water, the aqueous solution is
acidified (pH 2.5), and extracted with ethyl
acetate (3 x 20 ml.). The combined organic extract
is dried over sodium sulfate, filtered, and con-
centrated to give 599 mg. of N-[(l,1-dimethyl-
ethoxy)carbonyl]-L-phenylalanine]glycine.
To a mixture of the above acid (plus an
added amount from a small scale run) (644 mg., 2.0
mmole) and [(lS)-1-(cyclohexylmethyl)-2-hydroxy-2-
1 31 07q2
HA375a
-63-
[l-[(phen~lmethoxy)methyl]-lH-imidazol-2-ylJ-
ethyl]carbamic acid, l,l-dimethylethyl ester, slow
isomer from Example 6(d) (887 mg., 2.0 mmole) in
dimethylformamide ~8 ml.) at 0 is added l-hydroxy
ben~otriazole hydrate (306 mg., 2.0 mmole), diiso-
propylethylamine (731 ~1., 4.~ mmole), and finally
dicyclohexylcarbodiimide (420 mg., 2.0 mmole). The
reaction mixture is stirred for 2 hours at 0 and
then overnight at room temperature. The next day,
the precipitated dicyclohexyl urea is filtered
off, and the filtrate is concentrated. The ~
residue is taken up in ethyl acetate (75 ml.) and
washed sequentially with water (2 x 25 ml.),
saturated aqueous sodium bicarbonate (2 x 25 ml.),
and saturated aqueous sodium chloride (25 ml.),
dried over sodium sulfate, a~d concentrated to
give 838 mg. of crude product. Repeated chroma-
tographic purifications (silica gel, eluting with
19:1 chloroform:methanol) gives 113 mg. of N-[(lS)-
1-(cyclohexylmethyl)-2(R)-hydroxy-2-[1-[(phenylmethoxy)-
methyl]-lH-imidazol-2-yl]ethyl]-N2-[N-[(1,l-dimethyl-
ethoxy)carbonyl]-L-phenylalanyl]glycinamide.
TLC (silica gel; chloroform:methanol:
acetic acid, 9:1:1) Rf - 0.3.
Anal. calc'd. for C36H49N5O6:
C, 66.81; H, 7.62; N, 10.81
Found: C, 64.49; H, 7.41; N, 10.02.
c) N-~(lS)-l-(Cyclohex~lmethyl)-2(~)-hydroxy-2-
tlH-imidazol-2-yl)ethyll-N2-[N-[(l,l-dimethyl-
ethoxy)carbonyll-L-phenylalanyl]glycinamide~
acetate salt
20% Palladium hydroxide on carbon catalyst
(100 mg.) is weighed in a side arm flask and flushed
with hydrogen using a balloon. The glycinamide product
~ 1 0 7 q 2
HA375a
-64-
from part (b) (118 mg., 0.2 mmole) is added as a
2 ml. methanol solution. This is followed by the
sequential addition of methanol (6 ml.), water
(2 ml.) and 10% aqueous hydrochloric acid (145 ~1.).
~he reaction mixture is stirred at room temperature
for several days. The catalyst is removed by
filtration and ~he filtrate is concentrated to
give a crude product which after repeated
~hromatographic separations (silica gel, eluting
with chloroform:methanol:water:acetic acid,
90:20:2.5:1) yields 9.5 mg. of N-[(lS)-l-(cyclo-
hexylmethyl)-2(R)-hydroxy-2-(~H-imidaæol-2-yl)ethyl]-
N2-[N-[(l,l-dimethylethoxy)carbonyl]-L-phenyl-
alanine~glycinamide, acetate salt;
m.p. 99 - 100; [~D = ~7-0 (c = 0.1, methanol).
TLC (silica geli chloroform:methanol:water:acetic
acid, 90:20:2.5:1) Rf = 0.27.
Anal calc~d for C28H41N55 0 7 C2 4 2 2
C, 60.16; H, 7.86; N, 11.92
~0 Found: C, 60.06; ~, 7.64; N, 11.93.
Exam~le 10
N2-rN-r(l,l-Dimethylethoxy~L~arbonvl]-L-~henylalanylL-
N-[(S)-1-r(R~-hvdroxy(2-thia~olyl)methyl]-3-methyl
butvl]-L-histidinamide, acetate salt
~5 a) (2S~-2-rr(1,l-Dimethylethox~ bonyl]aminol-
-methyl-1-(2-thiazolyl2-1-pentanol
2.5N n-Butyllithium in hexane (8 ml.) is
added to a solution containing thiazole (1.7 g.,
~0 mmole) in dry tetrahydrofuran (60 ml.) cooled
to -70 under argon. After stirring a short time,
the solid material begins to come out of
1 3 i 0792
HA375a
-65-
solution. The reaction remains heterogeneous after
35 minutes at -40 to -35. After cooling the
reaction to -50, a solution containing N-[(1,1-
dimethylethoxy)carbonyl]-L-leucinal (2.2 g.,
10 mmole) in tetrahydrofuran (5 ml.) is added.
After 90 minutes the reaction is warmed to -10
and quenched with saturated ammonium chloride (10 ml.).
The reaction mixture is extracted into ether and
the organic extract is rinsed with water and brine,
dried (MgSO4) and concentrated in va~uo to give
1.8 g. of amide product. Two flash chromato-
graphies on first 100 g. and then 150 g. o silica
gel (LPS-l) eluting with petroleum ether:acetone
(20:1) fails to remove a minor impurity from the
co-eluting desired diastereomeric product
mixture. A total of 1.2 g. of (2S)-2-[[1,1-
dimethylethoxy)carbonyl]amino]-4-methyl-1-(2-
thiazolyl)-l-pentanol is obtained. TLC (silica
gel; petroleum ether:acetone, 4:1) Rf = 0.37.
[~]D = -38.6 (c = 1, chloroform).
b) [(l,l-Dimethylethoxy)carbonYl~-N -(2,4-
dinitro~henyl)-N-[(S~ (R~ydroxy(2-thiaæol~l)-
methyl]-3-methvlbutyll-L-histidinamide
A solution of the product from part (a)
(1.5 g., 5 mmole) dissolved in ethyl acetate
(40 ml.) is cooled in an ice water bath under
argon. After saturating with dry HCl gas, the
reaction is kept cold or 30 minutes. After
removal of the ethyl acetate ln vacuo, trituration
of the residue gives 1.45 g. of crude bis
hydrochloride salt.
1 3 1 079~
HA375a
-6~-
This bis hydrochloride salt (860 mg.,
3.15 mmole), N-[(l,l-dimethylethoxy)carbonyl]-
Nl-(2,4-dinitrophenyl)-L-histidine (1.38 g.,
3.15 mmole), and l-hydroxybenzotria7O1e hydrate
(482 mg., 3.15 mmole) are stirred in tetrahydro-
furan ~30 ml.) under argon; cooled in an ice water
bath, and treated with N-methylmorpholine (637 mg.,
6.3 mmole) followed by dicyclohexylcarbodiimide
(650 mg., 3.15 mmole). The reaction is stirred in
the ice bath for a~ hour, refrigerated overnight,
and then filtered. The filtrate is diluted with
ethyl acetate. The organic extract is rinsed with
water, saturated sodium bicarbonate, and brine,
dried (MgSO4), and concentrated ln vacuo to give
1.82 g. of crude product. Flash chromatography on
silica gel (170., LPS-l) eluting with chloroform:
methanol (20:1) yields 0.46 of fast moving isomex
(S), 0.57 g. of a mixture of isomers, and 0.27 g.
of slow moving isomer (R). Rechromatographing the
mixture ~raction gives a total of 0.54 g. of
(S) isomer and 0.80 g. of r(l,l-dimethylethoxy)-
carbonyl]-Nl-(2,4~dinitrophenyl)-N-[(S)-1-[(R)-
hydroxy(2-thiazolyl)methyl]-3-methylbutyl]-L-
histidinamide; m.p. 82 - 107. TLC (silica gel;
chloroform:methanol 20:1) Rf = 0.15. [~]D =
+12.7 (c = 1, methylene chloride).
c) N -rN-[(1,1-Dimethylethoxy2carb~ -L-p_enyl-
alanyll-N-[(S)-l-[(R)-hydroxY(2-thiazolVl)methYll-
3-methylbutyll-Nl-(2,4-dinitrophenyl)-L-histidlnamide
A solution containing the (R) hydroxy isomer
product from part (b) (510 mg., 0.84 mmole) in
1 3 1 ~7~
HA375a
-67~
ethyl acetate (25 ml.) is cooled in an ice water
bath and saturated with dry HCl. After stirring
for 60 minutes, the solvent is removed in vacuo to
yield 436 mg. of Nl-(2,4-dinitrophenyl)-N-~(S)-l-
S [(R)-hydroxy(2-thiazolyl)me~hyl]-3-methylbutyl]-
L-histidinamide, hydrochloride salt.
This crude hydrochloride salt (436 mg.,
O.46 mmole), N-[(1,l-dimethoxyethoxy)carbonyl]-L-
phenylalanine (122 mg., O.46 mmole), and 1-hydxo~y-
benzotria~ole hydrate (70 mg., 0.46 mmole) are _-
stirred in tetrahydrofuran (10 ml.) under argon,
cooled to 0 and treated with N~methylmorpholine
(118 mg., l.lS mmole) followed by dicyclohexyl-
carbodiimide (95 mg., 0.46 mmole). The reaction
is allowed to warm to room temperatuxe overnight,
then filtered and diluted with ethyl acetate and
ether. The organic extract is rinsed with water,
saturated sodium bicarbonate, and brine, dried
(MgSO4), and concentrated 1n vacuo to give 360 mg.
~0 of crude product. Flash chromatography on silica
gel (40 g., LPS-l) eluting with chloroform:methanol
(20:1) yields 250 mg. of N2-[N-[(1,1-dimethylethoxy)-
carbonyl]-L-phenylalanyl]-N-[(S)-l-[(R)-hydroxy-
(2-thia~olyl)methyl]-3-methylbutyl]-Nl-(2,4-
~5 dinitrophenyl)-L-histidinamide, m.p. 100 ~ 115;
[~]D = +25.5 (c = 1, chloroform). TLC(silica gel;
chloroform:methanol, 10:1) Rf = 0~32.
~n~l~ calc d. for C35H42N8OgS 1.5 H2O:
C, 54.04; H, 5.R3; N, 14.41; S, 4.12
Found: C, 53.99; H, 5.59, N, 13.97; S, 4.18.
1 3 ~ 0792
HA375a
-68-
d) N2-[N-[(l,l-Dimethylethoxv)carbonyl]-L-phenyl-
alanyl]~N-L~ 1-L(R)-hydroxy(2-thiazoly~)methyl]-3-
methylbutyll-L-histidinamide, acetate salt
Mercaptoacetic acid (1.15 g., 12.5 mmole) i5
added to a solution of the product from part (c)
(243 mg., 0.324 mmole) in dimethylformamide (3 ml.
under argon. After 2 hours at room temperature,
the reaction is extracted into a mixture of ethyl
acetate (40 ml.) and ether (20 ml.), rinsed with
seven 10 ml. portions of 10% a~ueous sodium
carbonate, three 10 ml. portions of water, and
brina. The organic extract is dried (MgSO4) and
concentrated ln vacuo to 0.26g. of crude
product. Flash chromatography on silica gel
(25 g., LPS - 1) eluting with chloroform:
methanol:water:acetic acid (90:15:1:0.5) gives
112 mg. of product. This material is dissolved in
0.5% aqueous acetic acid ~20 ml.), millipore
filtered, and lyophillized to give 99 mg. of N2-
[N-[(l,l-dimethylethoxy)carbonyl]-L-phenylalanyl]-
N-[(S)-l-[(R)-hydroxy(2-thiazolyl)methyl]-3-methyl-
butyl]-L-histidinamide, acetate salt; m.p. 89-115;
[~]D = -30.2 (c = 0.5, methanol). TLC (silica
gel; chloroform:methanol:water:acetic acid,
90:15:1:0.5) Rf = 0.21.
Anal. calc'd for C29H40N6O5S 0.5 C2 4 2 2
C, 56.94; H, 7.01; N, 13.28; S, 5.07
Found: C, 57.00; H, 6.63; N, 13.28; S, 4.78.
I ~` 1 07"~
HA375a
-69-
Example 11
N-[(S)-2-CYclohexyl--l-[(R~-hydroxy(lH--lmidazol-2
vl)methyl]ethyl]-N2-[N-(pyrrolidinYlcarbonyl)-L
~henylalanyl]~L-histidinamide, dihydrochloride
a) N-~1-Pyrrolidinylcarbonyl)-L-phenylalanine,
methvl ester
N-Methylmorpholine (11 ml., 100 mmole) and
phosgene (101 ml. of 12% solution in benzen~,
80 mmole) are added rapidly (dropwise) to a
solution of pyrrolidine (3.34 ml., 40 mmole) in
methylene chloride (200 ml.) at -30 under argon.
The resulting mixture is stirred for one hour at
-30, then for one hour as the temperature warms
to ~5, after which the mixture is concentrated ln
vacuo at 25. The residue is redissolved in
methylene chloride. N-Methylmorpholine (13.2 ml.,
120 mmole) followed by L-phenylalanine, methyl
ester, hydrochloride (8.63 g., 40 mmole) are then
added. The mixture is stirred overnight under
argon at 25, after which it is concentrated to
dryness. The residue is dissolved in ethyl
acetate, washed sequentially with water, lN
hydrochloric acid, and saturated aqueous sodium
bicarbonate solution, dried (MgS04), and concenta-
~5 ted. The residue is chromatographed on silica gel(Merck, 300 g.) eluting with benzene:acetic acid
(6:1). Fractions containing the product (Rf =
0.4) are combined and concentrated. The residue
(1.5 g.) is crystallized from ethyl acetate/hexane
to give 1.19 g. of N-(1-pyrrolidinylcarbonyl)-L-
phenylalanine, methyl ester; m.p. 93 - 95;
~ 3 ~ 0792
HA375a
-70-
[~]D = -19.4 (c = 1, methanol~.
b) N-(1-Pyrrolidinylcarbonyl?~L-E~henylalanine
A mixture of the methyl ester product from
part ~a) (1.187 g., 4.3 mmole),aqueous lN sodium
hydroxide solution (5.15 ml., 5.15 mmole), and
methanol (17 ml.? is stirred at 25 for 4 hours,
after which it is concentrated ln vacuo. lN
Hydrochloric acid and ethyl acetate are added to
the residue, and the mixture is extracted with
ethyl acetate. The extract is dried (MySO4) and
concentrated to gi~e N-(l-pyrrolidinylcarbonyl)-L-
phenylalanine; [~]D = -12.8 (c = 1, methanol).
c) (~R,~S)-~ Am ~ };IIE~hen lmethoxy)me~hyl]-
lH~imidazol-2-yl]cyclohexanepro~anol
A solution of [(lS)-l-(cyclohexylmethyl)-2-
hydroxy-2-[1-[(phenylmethoxy)methyl]-lH-imidazol-
~-yl]ethyl]carbamic acid, 1,1-dimethylethyl ester
(slow moving isomer) (3.92, 8.83 mmole) [prepared
as described in Example 6(d)] in ethyl acetate
(200 ml.) is cooled to 0 and HCl gas is bubbled
through the solution for 30 minutes. The mixture
is then stirred or 3.5 hours as it warms to room
temperature, after which it is concentrated ln
vacuo to give 3.56 g. of (aR,~S~-~-amino-~-[1-
[(phenylmethoxy)methyl]-lH-imidazol-2-yl]cyclohexane-
propanol as a white powder.
d) [(l,1-Dimeth~lethoxy?carbonyl~-N-[(lS,2R)-l-
(cyclohexylmethyl)-2-hydroxy-2-[1- r ( phenylmethoxy?-
methYl]-lH-imidazol-2-yl]-N3-[(phenylmethoxy?methyl]-
30 L-histidinamide -~
Triethylamine (2.06 ml., 14.7 mmole) and
dicyclohexylcarbodiimide (1.52 g., 7.35 mmole)
~ 3 1 07~2
HA375a
-71-
are added to a solution of the product from part
(c) (3.06 g., 7.35 mmole), l-hydroxybenzotriazole
hydrate (1.13 g., 7.35 mmole) and N-(l,l~dimethyl-
ethoxy)carbonyl]-l-[(phenylmethoxy)methyl] L-
histidine (2.76 g., 7.35 mmole) in tetrahydrofuran
(20 ml.). The mixture is stirred for 18 hours at
25, after which it is filtered. The filtrate is
diluted with ethyl acetate, washed with saturated
sodium bicarbonate solution, dried (MgS04), and
concentxated. The residue (4.92 g.) is
chromatographed on silica gel (Merck) eluting ~`
with ethyl acetate:pyridine:acetic acid:water
(80:20.6:11) to give as the major product 3.98 g.
of [~l,l-dimethylethoxy)carbonyl~-N-[(lS,2R)-l-
(cyclohexylmethyl)-2-hydroxy 2-[1-[(phenylmethoxy)-
methyl]-lH-imidazol-2-yl]-N3-[(phenylmethoxy)methyl]-
L-histidinamide; [~]D ~ -6.1 (c = 1.8, methanol).
e) N-[(lS,2R)-l-(Cyclohexylmethyl)-2-hydroxy-2-
[l-[(phenylmethoxy)methyl]-lH-imidazol-2-yl]ethyll-
N3-[(phenylmethoxy)methyl]-L-histidinamide
A solution of the product form part (d)
(3.88 g., 5.53 mmole) in ethyl acetate (200 ml.)
is cooled to 0 in an ice bath and HCl gas is
bubbled through the solution for 30 minutes. The
resulting mixture is then stirred for 2.5 hours as
it warms to 25, after which it is concentrated
to a small volume. The resulting white precipitate
is collected to give 3.33 g. of N-[(lS,2R)-l~(cyclo-
hexylmethyl)-2-hydroxy-2-[1-[(phenylmethoxy~methyl]-
lH-imidazol-2-yl]ethyl]-N3~[(phenylmethoxy)methyl]-
L-histidinamide as a white powder;
1 3 1 0 7~2
HA375a
-72-
m.p. 143 ~ 157; [~]D = +18-4 (c = 1.0,
methanol).
f) N-[(S)-2-Cyclohexyl-l-[(R~hydroxy-[l-
[(ph~nylmethoxv)methyl]~lH-imidazol-2~yl]methyl]-
ethyl]-N2-[N-(pyrrolidinYlcarbonyl)~L-phenYlalanyl]
N -[(phenylmethoxy)methyl]-L-histidinamide
Triethylamine (0.84 ml., 6.0 mmole) followed
by dicyclohexylcarbodiimide (453 mg., 2.2 mmole)
are added to a mix~ure of N~ pyrrolidinylcar-
bonyl)-L-phenylalanine (577 mg., 2.2 mmole),
l-hydroxybenzotriazole hydrate (337 mg., 2.2 mole);
and the L-histidinamide product from part (e)
(1.47 g., 2.0 mmole) in tetrahydrofuran (8 ml.).
The resulting mixture is stirred overnight as it
warms to 25. It is then filtered and the
filtrate is diluted with ethyl acetate, washed
with saturated sodium bicarbosate solution, dried
(MgSO4~, and concentrated. The residue (1;7 g.)
is chromatographed onisilica gel eluting with
ethyl acetate:pyridine:acetic acid:water
(30:20:6:11) to give as the major product 1.46 g.
of N-[(S)-~-cyclohexyl-l-[(R~-hydroxy-[l-~(phenyl-
methoxy)methyl]-lH-imida~ol-2-yl]methyl]ethyl~-N2-
[N-(pyrrolidinylcarbonyl)-L-phenylalanyl]-N3-[(phenyl-
methoxy)methyl]-L-histidinamide; [~]D = ~14.2
(c=0.8, methanol).
g) N-[(S)-2-Cyclohexyl-1-[(R)-hvdroxy-(lH-imidazol-
~-yl~methyl]ethyl]-N2-[N-(pyrrolidinYlcarbonyl~-L-
henylalanyl~-L-histidinamide, dihydrochloride
A mixture of the product from part (f) (1.41
g., 1.60 mmole), lN hydrochloric acid (3.36 ml.,
1 3 1 07q2
HA375a
-73-
3.36 mmole), and 20% palladium hydroxide on carbon
catalyst (300 mg.) in methanol (25 ml.) is stirred
under a stream of hydrogen for 24 hours. It is
then filtered and concentrated. The residue is
lyophillized from water to give 1.0 g. of N-[(S)-
2-cyclohexyl-1-[~R)-hydroxy(lH-imidazol-2-yl)-
methyl]ethyl]-N2-[N-(pyrrolidinylcarbonyl)-L-
phenylalanyl]-L-histidinamide, dihydrochloride as
a fluffy white powder; m.p. (168) 175 - 180;
[~]D = -44.8 (c = 0.9, methanol). TLC (silica
gel; ethyl acetate:pyridine:acetic acid:water,
40:20:6:11) R~ = 0.28.
32 44 8 4 2.2HC1 3.5 H20:
C, 51.38; H, 7.17; N, 14.98; Cl, 10.43
Found: C, 51.18; H, 7.09; N, 14.98; Cl, 10.48.
Exam~le 12
N-t(S)-2-Cyclohexyl-l-r(R)-hydroxy(lH-imidazol-2-
vl)methyllethyl]-N2-[N-~ dimethylethyl)amino]
carbonyll-L-~henylalanvl]-L-histidinamide,
dihydrochloride
a) N-[(4-Nitrophenoxy~carbonyl]-L~henylalanine,
methyl ester
N-Methylmorpholine (2.2 ml., 20 mmole)
followed by 4-nitrophenyl chloroformate (2.01 g.,
10 mmole) are added to a suspension of L-phenyl-
alanine, methyl ester, hydrochloride (2.15 g.,
10 mmole) in methylene chloride (40 ml.) at -30.
The resulting mixture is stirred at -30 for 15
minutes, then for 15 minutes at 25, after which
it is washed sequentially with lN HCl and
saturated aqueous sodium bicarbonate solution,
13~01q2
HA375a
-74-
dried, and concentrated. The residue (2.96 g.) is
crystallized from acetonitrile to give 1.22 g. of
N-[(4-nitrophenoxy)carbonyl]-L-phenylalanine,
methyl ester; m.p. 130 - 131, [~]D = ~88
(c = 1.5, chloroform). Tlle mother liquor is
chromatographed on silica gel (90 g.) eluting with
benzene:ethyl acetate (9:1) to give an additional
760 mg. of product.
b) N-[[~l,l-Dimethylethyl)aminolcarbonYll-L-
phenylalanine, methyl ester _.
l,l-Dimethylethyl amine (0.56 ml., 5.4 mmole)
is added to a solution of the product from part (a)
(1.48 g., 4.3 mmole) in toluene (21 ml.) at 0.
The resulting mixture is stirred for 24 hours as
it warms to 25, after which it is concentrated ln
vacuo. The residue is dissolved in ethyl acetate
and the solution is washed sequentially with lN
HCl solution, saturated aqueous sodium bicarbonate
solution, and saturated potassium carbonate
solution. The organic phase is filtered and the
filtrate is washed with aqueous potassium
carbonàte solution until the washes are colorless.
The or~anic extract is dried (MgSO4) and
concentrated in vacuo. The residue (1 17 g.) is
~5 crystallized from ethyl acetate/hexane to give
850 mg. of N-[[(l,l-dimethylethyl~amino]carbonyl]-
L-phenylalanine, methyl ester; m.p. 84-86;
[~]D = +24.4 (c = 0.9, methanol).
c) N-[[~ Dimet~y~l
Phenvlalanine
A mixture of the methyl ester product from
part (b) (838 mg., 3.0 mmole) and aqueous lN
I ~ 1 07q2
HA375a
-75-
sodium hydroxide solution (3.3 ml., 3.3 mmole) in
methanol (3 ml.) is stirred for 2 hours at 25,
after which it is concentrated in vacuo. The
residue is dissolved in water and washed with
ethyl acetate. The agueous layer is made acidic
by the addition of lN HCl solution and extracted
with ethyl acetate. The extract is dried (MgSO4)
and concentrated to give 603 mg. of N-[[(l,1-
dimethylethyl)amino]carbonyl]-L-phenylalanine.
[a]D = +39.6 (c = 0.7, methanol).
d) N-r(S)-2-C~clohexyl-1-[(R)-hydroxy[l-[(phenyl-~-
methoxy~methvl~ imidazol-2-y~lmethyl]ethyl]-N2-
[N-rU~ dimethylethyl2a ino~c rbonyll L-phenyl-
alanvl]-N3-~phenylmethoxy~methyl]-L-histidinamlde
Triethylamine (0.84 ml., 6.0 mmole) and
dicyclohexylcarbodiimide (453 mg., 2.~0 mmole) are
added to a mixture of N-[(lS,2R)-l-(cyclohexyl-
methyl)-2-hydroxy-2-[1-[(phenylmethoxy)methyl]-lH-
imidazol-2-yl]ethyl]-N3-[(phenylmethoxy)methyl]-L-
histidinamide (1.47 g., 2.0 mmole) [prepared as set
forth in Example ll(e)], the product from part (c)
(582 mg., 2.20 mmole), and l~hydroxybenzotriazole
hydrate (337 mg., 2.20 mmole) in tetrahydrofuran
(8 ml.) at 0. The resulting mixture is stirred
~5 overnight as it warms to 25, after which it is
filtered. The filtrate is diluted with ethyl
acetate, washed with saturated sodium bicarbonate
solution and brine, dried (~gSO4), and concentrated.
The residue is flash chromatographed on silica gel
(Merck) eluting with ethyl acetate:pyridine:acetic
acid:water (80:20:6:11) to give as the major
~ 3 1 0792
~76- HA375a
product 1.48 g. of N-[(S)-2-cyclohexyl-l~[(R)-
hydroxy[l-[(phenylmethoxy)methyl] lH-imidazol-2-
yl]methyl]ethyl]-N2-[N-[~(l,l-dimethylethyl)amino]-
carbonyl]-L-phenylalanyl]-N3-[(phenylmethoxy)methyl]-
L-histidinamide. [~]D = ~3-5 (c = 9, methanol).
e) N-[(S)-2-CyclohexYl-l-[(R)-hYdroxY(lH-imidazol-
2-vl)methyl]ethyll-N2-[N-L[(l,l-dimethylethyl)amino]
carbonyl]-L-phe~ylalanyl~-L-histidinamide,
dihydrochloride
A mixture of the product from part (d) ~-
(1.43 g., 1.69 mmole) and 20% palladium hydroxide
on carbon catalyst (300 mg.) in methanol (25 ml.)
is stirred under a stream of hydrogen for 24 hours,
after which it is filtered and concentrated. The
residue is lyophilized from water to give 1.064 g.
of N-[(S)-2-cyclohexyl-1-[(R)-hydroxy(lH-imidazol-
2-yl)methyl)ethyl]-N2-[N-[[(l,l-dimethylethyl)amino~-
carbonyl]-L-phenylalanyl]-L-histidinamide, dihydro-
chloride as a white solid; m.p. tl75) 178-185;
[~]D = -27.7~ (c = 0.9, methanol). TLC (silica gel;
ethyl acetate:pyridine:acetic acid:water,
40:20:6:11) Rf = 0.27.
32 46 8 4 2.lHC1 2.5~2O:
C, 52.77; H, 7.35; N, 15.38; Cl, 10.22
Found: C, 52.56; ~, 7.36; N, 15.24; Cl, 10.37.
1 3 1 07q2
HA375a
-77-
Example 13
N-~(S)-2-Cyclohexyl-l-[(R)-hydroxy(lH-imidazol-2-
yl~methyllethyll-N2-[N-(cyclopentylcarbonyl~-L-
phenylalanyll-L-histidinamide~ dihydrochloride
a) N -(L-Phenylalanyl)-N-[(lS,2R)-2~cyclohexyl-1-
[hydxoxy[1~ enylmethoxy)methyll~lH-imidazol-2-yl
methyl]ethyl~-N3-l~h nYlmethoxY?methyl]-L-histidin-
amide, trihydrochloride salt
A solution of N2-[N-[(1,1-dimethylethoxy)-
carbonyl]-L-phenylalanyl]-N-[(S)-1-[(R)-hydroxy-
[l-(phenylmethoxy)methyl] lH-imidazole-2-yl]- --
methyl]-2-cyclohexyleth~1]-3'-[~phenylmethoxy)-
methyl]-L-histidinamide (7.63 g., 9.0 mmole~
[prepared as set forth in Example 7(a)] in ethyl
acetate (325 ml.) is cooled in an ice-water bath
under argon and then saturated with HCl gas. The
mixture is stoppered and stirred cold for 30
minutes, then the bath is removed and the mixture
is allowed to warm to 25 over 60 minutes.
Removal of the solvents ln vacuo followed by
drying of the colorless solid product in vacuo
gives 7.64 g. of crude N2-(L-phenylalanyl)-N-
[(lS,2R)-2-cyclohexyl-1-[hydroxy[1-[(phenylmethoxy)-
methyl]-lH-imidazol-~-yl]methyl]ethyl]-N3-[(phenyl~
~S mathoxy)methyl]-L-histidinamide as a trihydrochloride
salt.
b) N-[(S)-2-Cyclohexyl-I- r ( R)-hydroxy[l- r ( phenyl-
methoxy)methyl]-lH-imidazol-2-y~methyl]ethyl]-N2-
~N-(cyclo~entylcarbonyl)-L-phenYlalanYl]-N3-
[(Phenylmethoxy)methyl]-L-histidinamide
Triethylamine (0.63 ml., 4.5 mmole) and
dicyclohexylcarbodiimide (340 mg., 1.65 mmole)
1 3 1 ~7q2
HA375a
-78-
are added to a solution of the trihydrochloride
salt product from part (a) (1.34 g., 1~5 mmole),
l-hydroxybenzotriazole hydrate (252 mg., 1.65 mmole),
and cyclopentanecarboxylic acid (0.18 ml.,
1.65 mmole). The resulting mixture is stirred for
18 hours at 25 after which it is filtered. The
filtrate is diluted with ethyl acetate, washed
with saturated sodium bicarbonate solution, dried
(MgSO4), and concentrated. The residue (1.25 g.)
is chromatographed on silica gel (Merck) eluting
with ethyl acetate:pyridine:acetic acid:water
(80:20:6:11) to give as the major product 800 mg.
of N-[(S)-2-cyclohexyl-l-[(R)-hydroxy[l-[(phenyl-
methoxy)methyl]-lH-imidazol-2~yl]methyl]ethyl]-N2-
[N-(cyclopentylcarbonyl)-L-phenylalanyl]-N3-[(phenyl-
methoxy)methyl]-L-histidinamide [~]D = -8.1
(c = 1.04, methanol).
c) N-[(S~-2-Cyclohexyl-l-~(R)-hydroxy(lH-imidazol-
2-yl)methyl]ethyl]-N -~N-(cyclopentylcarbonyl)-L0 PhenYlalanvlL-_-hlstidinamide, dihydrochloride
A mixture of the product from part (b)
(740 mg., 0.87 mmole), 20% palladium hydroxide on
carbon catalyst (150 mg.), and 1.0 N hydrochloric
acid (1.83 ml., 1.83 mmole) in methanol (30 ml.)
~5 is hydrogenated under a slow stream of hydrogen
~or 18 hours. The mixture is then filtered and
concentrated to dryness. The residue is dissolved
in water and lyophilized to give N-[(S)-2-cyclohexyl-
l-[(R)-hydroxy(lH-imidazol-2-yl)methyl]ethyl]-N2
[N-(cyclopentylcarbonyl)-L-phenylalanyl]-L-histidin-
amide, dihydrochloride; m.p. (155) 174-177.
1 3 1 07~2
HA375a
-79-
~]D = -28.2~ (C = 0.97, methanol). TLC (silica
gel; ethyl acetate:pyridine:ac~tic acid:water,
40:20:6:11) Rf = 0.2S.
Anal- calc'd- for C33H45N7Q4 2-15 HCl 3-36 H20
C, 53.37; H, 7.31; N, 13.20; Cl, 10.26
Found: C, 53.37; H, 7.30; N, 13.31; Cl, 10.31.
Example 14
N-~(S)-2-Cyclohexyl-1-_[~R)-hydrox ~lH- midazol-?-
yl)methyllethYl]-N -~N-(3,3-dimethvl-1-oxobutyl)-L-
phenylalanyl]-L-histidinamide, dihy~_chloride
a) N- r (S~-2 Cyclohexyl-l-~(R)-hydroxy[l-[(phenvl-.
methoxY)methy~-lH-imidazol-2-ylLmethyllethyll-N2-
~N-(3,3-dimethyl-1-oxobu~yl)-L-phenylalanyl]-N3-
[(phenvlmethoxy)methyll-L-histidinamide
Triethylamine (0.63 ml., 4.5 mmole) followed
by dicyclohexylcarbodiimide (340 mg., 1.65 mmole~
are added to a solution of N2-(L-phenylalanyl)-N-
[(lS,2R)-2-cyclohexyl-1-[hydroxy[l-[(phenylmethoxy)-
methyl]-lH-imidazol-2-yl]methyl]ethyl]-N3-[~phenyl-
methoxy)methyl]-L-histidinamide, hydrochloride salt
(1.34 g., 1.5 mmole) [prepared as described in
Example 13(e)], l-hydroxybenzotriazole hydrate
(252 mg., 1.65 mmole~ and 3,3-dimethylbutanoic acid
(0.21 ml., 1.65 mmole) in tetrahydrofuran (5 ml.)
at 0. The resulting mixture is stirred for 18
hours as it warms to 25, after which it is
filtered. The filtrate is diluted with ethyl
acetate, washed with saturated aqueous sodium
~ bicarbonate solution, dried (MgS04), and
concentrated. The residua (1.23 g.) is purified
by flash chromatography on silica gel (Merck, 150 g.)
1 ~ 1 07q2
HA375a
--~0--
eluting with ethyl acetate:pyridine:acetic acid:
water (80:20:6:11) to give as the major product
730 mg. of N-[(5)~2-cyclohexyl-1-[(R)-hydroxy[l-
[(phenylmethoxy)methyl]-lH-imidazol-2-yl]methyl]-
ethyl]-N -[N-(3,3-dimethyl~l-oxobutyl)-L-phenyl-
alanyl]-N3-[(phenylmethoxy)methyl]-L-histidinamide;
[~]D = -11.5 (c=l, methanol).
b) N-[5S)-2-Cyclohexyl-l-L(RI-h~droxy(lH-imidazol-
2-vl)methylL~thYl]-N2-[N-(3,3-dimethyl-1-oxobutyl)-
0 L-phenylalanyl]-L-histidinamide~ dihydrochloride
A mixture of the product from part (a)
(660 mg., 0.8 mmole), 20% palladium hydroxide on
carbon catalyst (150 mg.) and 1.0 N hydrochloric
acid (1.7 ml., 1.7 mmole) in methanol (20 ml.) is
hydrogenated under a slow stream of hydrogen for
24 hours. The mixture is then filtered and
concentrated to dryness. The residue (540 mg.) is
dissolved in water and activated charcoal (50 mg.)
is added. The resulting mixture is filtered and
lyophillized to give N-[(S)-2-cyclohexyl-1-[(R)-
hydroxy(lH-imida7ol-2-yl)methyl]ethyl]-N2-[N-(3,3
dimethyl-l-oxobutyl)-L-phenylalanyl]-L-histidin-
amide, dihydrochloride; m.p. 158~184~;
[~]D = -32.4~ (c = 0.79, methanol). TLC (silica
~el; ethyl acetate:pyridine:acetic acid:water,
40:20:6:11) R~ = 0.27.
r C33H47N704 2.15 HCl 2.5 H20:
C, 54.35; H, 7.4~; N, 13.45; Cl, 10.45
Found: C, 54.16; H, 7.45; N, 13.59; Cl, 10.42.
~ 3 1 ~7~1~
HA375a
-81-
Example 15
(lS,2R) N-~l-(CvclohexylmethYl~-2-hydro2y-2-51H-
imidazol-~-yl~ethvl]-N2-~N-(4 morpholinylcarbonyl)-
L-~henvlalanyl]-L-histidinamide, trifluoroacetate
salt
a) N-(4-Morpholinylcarbonyl)-L-phenvlalanine,
methyl ester
Morpholine (1.1 ml., 12.5 ml.) is added to a
solution of N-[(4-nitrophenoxy)carbonyl]-L-phenyl-
alanine, methyl ester (3.44 g., 10 mmole). The
resulting mixture is stirred for 2 hours at 25,
then at 100 for 5 hours, after which it is concen-
trated ln vacuo. The residue is dissolved in
ethyl acetate and the solution is washed with
saturated potassium carbonate solution until the
washes are colorless. The organic extract is
dried (MgS04) and concentrated ln vacuo. The
residue is crystallized from ethyl acetate/hexane
to give 2.3 g. of N-(4-morpholinylcarbonyl)-L-
phenylalanine, methyl ester; m.p. 88 - 91;
[a]D = -30.8 (c = 0.6, methanol).
b) N-(4-Moroholinylcarb n ~ ~-L-phenylalanine
A mixture of the methyl ester product from
part ~a) (2.3 g., 7.8 mmPle) and agueous lN sodium
hydroxide solution (8.6 ml., 8.6 mmole) in methanol
(12 ml.~ is stirred for 5 hours at 25, after
which it is concentrated ln vacuo. The residue i5
dissolved in water and washed with ethyl acetate.
The extract is dried (MgS04) and concentrated to
give 2.2 g. of N-(4-morpholinylcarbonyl)-L-phenyl-
~ alanine; [~]D = -23.8 (c = 2, methanol).
1 3 1 o 7 r~ L
HA375a
-82
c) ~lS,2R)-N-[1-(Cyclohexylmethyl)-2-hydroxy-2-
[1-~(phenylmethoxy)methyl]-lH-imidazol-2-yll-N
[N-(4-morpholinylcarb n~-L-~henylalanyl]-N3-
[(phenylmethoxy)methyl]-L-histidinamide
Triethylamine (0.84 ml., 6.0 mmole) and
dicyclohexylcarbodiimide (435 mg., 2.20 mmole)
are added to a mixture of N-C(lS,2R)-l-(cyclo-
hexylmethyl)-2-hydroxy-2-[1-[(phenylmethoxy)-
methyl]-lH-imidazol-2-yl~ethyl]-N3-[(phenyl-
methoxy)methyl]-L-histidinamide (1.49 g.,
2.0 mmole) [prepared as set forth in Example ll(e)],
the product from part (b) ~612 mg., 2.20 mmole),
and l-hydroxybenzotriazole hydrate (337 mg.,
2.20 mmole) in tetrahydrofuran (8 ml.) at 0.
lS The resulting mixture is stirred for 18 hours as it
warms to 25, after which it is filtered. The
filtrate is diluted with ethyl acetate, washed
with saturated sodium bicarbonate solution and
brine, dried (MgSO4), and concentrated. The
residue is flash chromatographed on silica gel
(Merck) eluting with ethyl acetate:pyridine:acetic
acid:water (100:20:6:11) to give as the major
product 1.42 g. of (lS,2R)-N-[1-(cyclohexylmethyl)-
2-hydroxy-2-[1-[(phenylmethoxy)methyl]-lH-imidazol-2-
yl]-N2-[N-(4-morpholinylcarbonyl)-L-phenylalanyl]-
N3-[(phenylmethoxy)methyl]-L-histidinamide;
[~]D = -15.3 (c = 0.9, methanol).
d) (lS,2R~-N-[l-(CYclohexYlmethyl)-2-hvdroxy-2-
(lH-imidazol-2-yl)ethyl]-N -IN-(4-mor~holinylcar-
bonyl)-L-Phenylalanyll-L-histidinamide, trifluoro-
acetate salt
A mixture of the product from part (c)
(1.4 g., 1.6 mmole), 1.0 N hydrochloric acid
1 3 1 0 7, 2
HA375a
-83-
~3.55 ml., 3.55 mmole), and 20% palladium hydroxide
on carbon catalys~ ~300 mg.) in methanol (25 ml.)
is stirxed under a stream of hydrogen for 18 hours,
after which it is filtered and concentrated. The
residue (1.7 g.) is purified by preparative HPLC
~YMC S15 ODS column 20 x 500 mm., 25 ml/min of
56% agueous methanol containing 1~ trifluoroacetic
acid, W absorbance monitored at 215 nm.).
Fractions containins the major product (retention
1~ time 22 minutes) are combined and concentrated.
The residue is lyophilized from water to give 850 mg.
o~ (lS,2R)-N~[l-~cyclohexylmethyl)-2-hydroxy-2-
(lH-imidazol-2-yl)ethyl]-N2-[N-(4-morpholinyl~
carbonyl)-L-phenylalanyl]-L-histidinamide, trifluoro-
lS acatate salt as a white solid; m.p. (76) 89-112;
[~]D = -38.1 (c = 0.935, methanol). TLC (silica
gel; ethyl acetate:pyridine:acetic acid:water,
40:20:6:11) Rf = 0.19.
~nal. calc'd. for C32H44N8O5 2.2 C2 3 2 2
~0 C, 48.65; H, 5.52; N, 12.47; F, 13.95
Found C, 48.69; H, 5.60; N, 12.49; F, 14.09.
Exam~le 16
(lS,2R~-N2-~N-~(4-Methyl~ erazinyl)carbonylL-L-
~henylalanylL-N-tl-(cyclohexylmethyl)-2-h~droxy-2-
S ~lH-imida201-2-yl)ethyll-L-histidinamide,
trihydrochloride
a) N-r(4-Methyl~ zinyl)carbonyl]-L
henvlalanine, methyl ester
A solution of N-[(4-nitrophenoxy)carbonyl]-L-
phenylalanine, methyl ester (3.44 g., 10 mmole) in
toluene (40 ml.) is heated to reflux and l-methyl
1 3 1 0792
HA375a
-84-
piperazine (1.4 ml., 12.5 mmole) is added to the
warm solution. The mixture is stirred for 3 hours
as it cools to 25, after which it is concentrated
ln vacuo. The residue is dissolved in ethyl
acetate and washed with aqueous potassium carbonate
solution. The residue (3.85 g.) is crystallized
from ethyl acetate to give 2.33 g. of N-[(4-methyl-
1-piperazinyl)carbonyl]-L-phenylalanine, methyl
ester as an off white solid; m.p. 137 - 138~;
[~]D = ~35 9 (c = 1, methanol).
b) N-~(4-Methyl-l-Diperazin~l)carbonyll-L-
phenylalanine
A solution of the methyl ester product from
part (a) (2.29 g., 7.5 mmole) in methanol (12 ml.)
and 1.0 N sodium hydroxide solution (8.25 ml.,
8.25 mmole) is stirred for 3 hours at 25, after
which the methanol is removed ~n vacuo. The
residue is acidified by the addition of excess
1.0 N ~Cl solution and is applied to a cationic
exchange column (lO0 ml bed of AG 50 W-X2). The
column is eluted with water until the eluant is no
longer acidic and then is eluted with 2% aqueous
pyridine. Product containing fractions are pooled
and concentrated. The residue tl.0 g.) is crystal-
lized by trituration with refluxing ethyl acetateto give 860 mg. of N-[(4-methyl-1-piperazinyl)car-
bonyl]-L-phenylalanine as a crystalline solid;
m.p. 130 - 132.
l3107Q2
HA375a
-85-
c) (lS,2R)-N2~[N-~(4-MethYl-1-piperazinyl)car-
bonyl]-L-phenylalanyll-N-[l-(cyclohexYlmethyl)-
2-hydroxy-2-~ (phenylmethoxy)methyl~-lH-imida
~-vllethv~l]-N -[(~henylmethoxy~methyl]-L-histidin-
amide
Triethylamine (0.92 ml., 6.6 mmole) and
dicyclohexylcarbodiimide (453 mg., 2.2 mmole) are
added to a solution of N-[(lS,2R)-l-(cyclohexyl-
methyl)-2-hydroxy-2-[1-[(phenylmethoxy)methyl]-lH-
imida2O1-2-yl~ethyl]-N3-[(phenylmethoxy)methyl]-L-
histidinamide (1.5 g., 2~0 mmole) [prepared as -.-
described in Example ll(e)~, l-hydroxybenzotriaæole
hydrate (337 mg., 2.2 mmole), and the product from
part (b) (690 mg., 2.2 mmole) in dimethylfo~mamide
(8 ml.) at 0. The mixture is stirred for 18
hours at 25, after which it is filtered. Ethyl
acetate is added to the filtrate and the mixture
is washed with saturated aqueous sodium bicarbonate
solution and brine, dried (MgSO4), and concentrated.
~he residue is flash chromatographed on silica gel
(150 g., Merck) eluting with chloroform:methanol:
ammonium hydroxide (100:12.5:0.25) to give as the
major product 600 mg. of (lS,2R)-N2~[N-[(4-methyl-
l-piperazinyl)carbonyl]-L-phenylalanyl]-N-[l-
(cyclohexylmethyl~-2-hydroxy-2-[1-~(phenylmethoxy)-
methyl]-lH-imida~ol-2-yl]ethyl]-N3-[(phenylmethoxy)-
methyl]-L-histidinamide; [~]D = -21.5 (c = 1,
methanol).
1 3 ~ 0792
HA375a
-86-
d) (lS,2R)-N2-[N-[(4-Methyl-l-~iperazinyl)car-
bonyl]-L-phenylalanyll-N-[1-(cyclohexylmethyl~-2-
_droxy-2-(lH-imidazol-2~yl)ethyl]-L-histidinamide,
trihydrochloride
A mixture of the product from part (c)
(563 mg., 0.64 mmole~, 20% palladium hydroxide
on carbon catalyst (125 mg.), l.ON HCl solution
(2.12 ml., 2.12 mmole) and methanol (20 ml.) is
hydrogenated under a slow stream of hydrogen for
~0 hours, after which it is filtered and
concentrated. The residue is dissolved in water,
charcoal filtered, and lyophillized to give 442
mg. of (lS,2R)-N -[N~[~4-methyl-1-piperazinyl)-
carbonyl]-L-phenylalanyl]-N-[1-(cyclohexylmethyl)-
2-hydroxy-2-(lH-imidazol-2-yl)ethyl]-L-histidinamide,
trihydrochloride; m.p. 178 - 202; [~]D = ~ 54 4
(c = 0.97, methanol). TLC (silica gel; chloroform:
methanol:ammonium hydroxide, 100:25:1) Rf = 0.39.
33 47 94 3-3 HCl 4.0 ~2
C, 47.96; H, 7.11; N, 15.26; Cl, 14.16
Found: C, 47.96; H, 7.06; N, 15.32; Cl, 14.19.
Example 17
(lS,2Rt-N-rl-(Cyclohexylmethyl)-2-hYdroxy-2-(2-
thiazol~l)ethyll-N2-[N-[.(l,l-dimeth~Ylethoxy~car-
~5 bonyll-L-phenylalanyll~L hl idinamide, O.5
acetate salt
a) (S~-~- r r (l,l-DimethYlethoxY~carbonyl]amino~-
cyclohexane ~ acid
Platinum oxide catalyst (5 g.) is added to a
solution of N-[(l,l-dimethylethoxy)carbonyl]-L-
phenylalanine (120 g., 0.452 mole) in absolute
1 3 1 07~2
HA375a
-87-
ethanol (1 l.). The mixture is placed on a Parr
reduction apparatus at 50 lbs. pressure. The
absorption of hydxogen is rapid and the hydrogen
reservoir needs continued refilling. The reduction
proceeds overnight and after 20 hours is
completed. The mixture is filtere~ through Celite
and concentrated in vacuo to give 124.4 g. of (S)~
~-[[(l,1-dimethylethoxy)carbonyl]amino]cyclohexanepro-
panoic acid as a glassy solid colorless xesidue;
[~]D = ~9-5 (c = 1, methanol). TLC (silica gel;
toluene:acetic acid, 4:1) Rf = 0.62.
b) ~S)-~-[[(1,1-Dimethylethoxy)carbon~l~amlno]-
N-methox~-N-methvlcvclohexanepropanamide
The product from part (a) (22.6 g.,
83.3 m~ole) is dissolved in tetrahydrofuran (250
ml.) under a blanket of argon at 26. Carbonyl-
diimidazole solid (16.0 g., 98.7 mmole~ is added
in portions over one minute. Moderate gas
evolution begins shortly after the addition is
completed. The mixture remains colorless through-
out. ~he mixture is stirred for 30 minutes at 25
during which time it remains clear and colorless.
O,N-Dimethylhydroxylamine hydrochloride (11.5 g.,
118 mmole) is then added in a single portion
followed immediately by triethylamine (17.5 ml.,
125 mmole) in a single portion. Following the
triethylamine addition a white precipitate forms.
The mixture is stirred for 3 hours at 25, after
which it is poured into lN HCl (400 ml.) and
30 extracted with ether (3 x 200 ml.). The colorless -~
extracts are combined and washed with saturated
1 3 1 0792
HA375a
-88-
so~ium bicarbonate solution (2 x 200 ml.), dried
~MgSO~), and concentrated to give 24.2 g. of
(S)-~-[[~l,l-dimethylethoxy)carbonyl]amino]-N-
methoxy-N-methylcyclohexaneprQpanamide; [~]D =
-11.1 (c = 7, methanol).
c) (S)-~l-(Cyclohexylmethyl)-2-oxo-2-(7-thia-
2 olyl)ethYl]carbamic acid, l,1-dimethylethyl
ester
A 2 . 6 M solution of n-butyllithium (19.5 ml.,
4.78 mmole) is added a solution of thiazole (4.07 g~,
4.78 mmole) in tetrahydrofuran (80 ml.) at -60
undar argon. The reaction is stirred at -60 for
30 minutes. The product from part (b) (7.5 g.,
2.4 mmole) in tetrahydrofuran (15 ml.) is added
lS dropwise at -60 and the reaction mixture is
stirred until the temperature reaches -20 (about
40 minutes). The reaction is quenched with
saturated ammonium chloride (40 ml.) and the
product is extracted with ether (4 x 200 ml.).
The organic layer is washed with brine, dried
(NgSO4), filtèred and concentrated to yield 7.2 g.
of crude product. This material is purified by
filtration through a 60 g. pad of Merck silica
using a hexane:ethyl acetate (8:2) solvent system.
The ~iltrate is concentrated _ vacuo to yield 6.0 g.
of crystalline (S)-[l-tcyclohexylmethyl)-2-oxo-2-
(2-thiazolyl)ethyl]carbamic acid, l,1-dimethylethyl
aster; m.p. 64 - 69. TLC (silica gel; hexane:
ethyl acetate, 8:2) Rf = 0.45.
3~ Anal. calc'd. for C17H26N~S3 0~1 hexane
C, 60.93; H, 7.90; N, 8~08; S, 9.24
Found: C, 61.17; H, 8.13; N, 7.95; S, 8.97.
1 3 1 ~792
HA375a
-89-
d) (lS,2R)~ Cvclohexylmethyl~-2-hydroxy-?-
(2 ~hiazolyl)ethyllcarbamic acid, ~ dimethyl-
ethYl ester
The product from part (c) ~2.73 g., 8.07
mmol~) is dissolved in absolute ethanol (50 ml.)
and cooled to 5~. Sodium borohydride (0.6 g.,
16.14 mmole~ is added portionwise and the reaction
mixture is stirred for one hour, diluted with
ether (200 ml.), and ~uenched with lN HCl to pH l.
The organic layer is separated, washed twice with
water and brine, dried (MgSO4), and concentrated .-
n vacuo. The two isomers are separated by flash
chromatography on silica gel (Merck, 300 g.)
eluting with ethyl acetate:hexane (3:8). The
slower moving isomer is identified as the S,S
configuration and the faster moving isomer is
identified as (lS,2R)-[1-(cyclohexylmethyl)-2~
hydroxy-2-~2-thiazolyl)ethyl]carbamic acid,
l,l-dimethylethyl ester; [~]D = -30.72 (c = 0.55,
methanol). TLC (silica gel; ethyl acetate:hexane;
1:1) Rf = 0.70.
e) (aR,~S)-B-Amino-~-(2-thiazolyl)cyclohexanepro-
panol, dihydrochloride
The product from part (d) (0.8 g., 2.3 mmole)
is dissolved in ethyl acetate ~20 ml.) and HCl is
bubbled into the solution for lO minutes, after
which it is stirred at room temperature for 4
hours. The reaction mixture is concentrated to
give (aR,~S)-~-amino-a-(2-thiazolyl)cyclohexane-
propanol, dihydrochloride as a white solid.
~ 3 1 07S2
HA375~
--90--
f3 N-~N-[(1,1-Dimethvlethoxv)carbonyl]-L-phenyl~
alanyll-1~-(2,~-dinitrophenyl)-L-histidine
2,4-Dinitrofluorobenzene (4.62 ml.,
36.74 mmole) is added to a solution of N-[N-
[(l,l-dimethylethoxy)carbonyl]-L-phenylalanyl]-L-
histidine (12.4 g., 30.8 mmole) in aqueous sodium
bicarbonate (lM,82 ml.) and methanol (103 ml.).
After stirring the solution for 2.5 hours, an
additional amount of 2,4-dinitrofluorobenzene
(0.6 ml., 4.77 mmole) is added. At the end of a
total of 4 hours of reaction time, the mixture is
acidified with aqueous hydrochloric acid (lN, 51.3
ml.) to a pH of 3.9 and then diluted with water
(500 ml.). The separated solid is filtered and
washed with water. This solid (16.6 g.) is then
crystallized from hot ethyl acetate. Dicyclohexyl-
amine (1.9 ml.) is added and the crystallized salt
is filtered and the free acid is regenerated by
acidification to give 1.008 g. of N-[N-[(l,1-
dimethylethoxy)carbonyl]-L-phenylalanyl]-1'-(2,4-
dinitrophenyl)-L-histidine; m.p. 180 - 182;
[~]D = +5.2 (c = 1.4, acetic acid).
g) ~lS,2R~ __[1-(CyclohexYlmethYl)-2-hydroxy-
2-(2-thiazolyl ? ethyll~N2-rN-r~ m~5hylethoxy)-
carbon~l~-L-~henylalanvl]-1'-(2,4-dinitrophenyl~
L-histidinamide
.. .. _
N-Methylmorpholine (0.23 ml., 2.1 mmole) is
added to a solution of the crude dihydrochloride
salt product from part (e~ (0.53 g., 1.5 mmole),
N-[N-[(1,1-dimethylethoxy)carbonyl]-L-phenyl-
alanyl]-1'-(2,4-dinitrophenyl)-L-histidine
1 3 1 0792
HA375a
--91~
(O.85 g., 1.5 mmole), and 1 hydroxybenzotriazole
hydrate (0.2 g., 1.5 mmole) at 0 under argon.
Dicyclohexylcarbodiimide (0.31 g., 1.5 mmole) is
finally added and the reaction is kept at 0
overnight, diluted with ethyl acetate (300 ml.),
and filtered. The filtrate is washed with water,
s~turated sodium bicarbonate solution, and brine,
dried ~gSO4), filtered and concentrated 1n vacuo
to yield 1.4 g. of crude product. Chromatography
on silica gel (Merck, 200 g.) eluting with
methanol:chloroform (5:100) gives 0.8 g. of
(lS,2R)-N-[l-(cyclohexylmethyl)-2-hydroxy-2-
(2-thiazolyl)ethyl]-N2-[N-[(l,l-dimethylethoxy)-
carbonyl]-L-phenylalanyl]-1'-(2,4-dinitrophenyl)-
1~ L-histidinamide; [~]D = -15.69 (c = 0.51,
methanol). TLC (silica gel; methanol:chloroform,
5:85) Rf = 0.25.
h) (lS,2R)- ~ exvlmet~y~ Ydroxy-2-
(2-thlazolyllethyl~-N -[N-~(l,l-dimethylethoxy2-
car~g~ lalanyll-L-h~stldinamide,
0.5 acetate salt
The product from part (g) (0.8 g., 1.0
mmole) is treated with thiolacetic acid (2.8 ml.)
in dimethylformamide (4 ml.) at room temperature.
After 2 hours, the reaction mixture is
concentrated ln vacuo, diluted with ethyl acetate
(200 ml.) and ether (100 ml.), washed with
saturated sodium bicarbonate solution (twice) and
brine (twice), dried (MgSO4), and concentrated
ln vacuo to give 0.8 g. of crude product. This
material is chromatographed on silica gel (Merck,
1 3 1 07q~
HA375a
-92-
200 g.) eluting with methanol:chloroform (5:100)
to remove the (S,S) isomer contamlnant. The
solvent system is changed to chloroform:methanol:
water:acetic acid (90:10:1:0.5) to elute the
desired ~S,R) isomer. The solution is concentra~
ted ln vacuo, dissolved in 2% aqueous acetic acid,
filtered (millipore), and lyophilliæed to give
O . 15 g . of ( 1 S, 2R ) -N- [ 1- ( cyclohexylmethyl)-2-
hydro~y-2-(2-thiazolyl)e~hyl]-N2 [N-[(l,l-dimethyl-
ethoxy)carbonyl]-L-phenylalanyl]-L-histidinamide,
0.5 acetate salt as a light yellow powder; m.p.
110 - 115; ~]D = -32.75 (c = 0.4, methanol).
TLC (silica gel; methanol:chloroorm, 1:9) Rf = O.29.
Anal. calc'd- for C32H44N65S 0 5 C2~I42 H20
C, 58.90; H, 7.19; N, 12.49; S, 4.76
Found: C, 58.69; H, 6.94; N, 12.38; S, 4.92.
Example 18
N-[(S)-1-~HydroxY(lH~imidazol-4-yl)methyl]-3-methyl~
butvl]-N2-rN-[~-dimethylethoxy)carbonyl]-L-
Dhenylalanyl]-L-histidinamide, 0.3 acetate salt
a) 1-[(PhenylmethoxY)methyl]-2-(phenylthio)-lH-
imidazole
A solution of n-butyllithium (3.8 ml.,
2.6 N in hexane) is added dropwise to a solution of
l-[(phenylmethoxy)methyl]-lH-imidazole (1.88 g.,
10 mmole) in tetrahydrofuran (35 ml.) at -60~ under
argon. After stirring for 2 hours, this solution
is added dropwise to a solution of diphenyldisulfide
~1.88 g., 10 mmole) ~t -60 and stirred at -50 to
30 ~60 for 2 hours. Saturated ammonium chloride `-
(20 ml.) is added dropwise at room temperature and
~3~07S2
HA375a
-93-
the crude product is extracted with ether (2 x
200 ml.), washed with brine, dried ~MgS04) and
concentrated in vacuo. Flash chromato~raphY on
.
silica gel (120 g., LPS-1) eluting with hexane:
ethyl acetate (7:3) yields 2.2 g. of l-[(phenyl-
methoxy)methyl]-2-phenylthio-lH-imidazole. TLC
(silica gel; hexane:ethyl acetate, 7:3) Rf = 0.21.
b) [(l,l-DimethYlethoxy)carbonYl]-N-methoxy-N-
methyl-L-leucinamide
N-[(l,l-Dimethylethoxy)carbonyl]-L-leucine
hydrate (10 g., 40 mmole) is dissolved in excess
toluene and concentrated to dryness. The residue
is dissolved in tetrahydrofuran (100 ml.) and
carbonyldiimida~ole (7.8 g., 48 mmole) is added in
a single portion. The mixture is stirred at room
temperature for 30 minutes. 0,N-Dimethylhydroxyl-
amine hydrochloride (4.3 g., 44 mmole) and txiethyl-
amine (6.2 ml., 44 mmole) are then added. The
resulting mixture is stirred at room temperature
~0 for 3 hours, after which it is poured into excess
lN hydrochloric acid. The mixture is extracted
- three times with ethyl acetate and the extracts
are washed once with lN hydrochloric acid and
twice with saturated sodium bicarbonate solution,
~5 dried (MgSO4), and concentrated to give 9.3 g. of
[(l,l-dimethylethoxy)carbonyl]-N-methoxy-N-methyl-
L-leucinamide as a colorless oil; [a]~ = -25.2
(c - 1.6, methanol).
c) (S)-r3-Methyl-1-[~3-(phenylmethoxy)methyl]-2-
(phenylthio)-3H-imidazol-4 yl]carbonyl]butyl]-
carbamic acid, l,l-dimethylethyl ester
Lithium dii~opropylamide is prepared 1n
1 3 1 07q2
HA375a
--9~L--
situ by the dropwise addition of n-butyllithium
(3.2 ml., 2.6 N in hexane) to a solution of
diisopropylamine (1.2 ml., 8.4 mmole) in tetra-
hydrofuran (24 ml.) under argon at -60 for
15 minutes. The product from part (a) (2.36 g.,
8 mmole) in tetrahydrofuran (20 ml.) is added
dropwise at -50 and stirred at -73 for 10
minutes. The product from part (b) (1.08 g.,
3.9 mmole) is added in tetrahydrofuran (6 ml.) at
--60 and stirred until the reaction reaches 0
(40 minutes). The reaction is then quenched by -
the addition of saturated ammonium chloride (10
ml.) and diluted with ether (400 ml.). The
organic layer is separated, washed with saturated
ammonium chloride and brine, dried (MgS04),
filtered, and concentrated in vacuo. The crude
product is purified by flash chromatography on
silica gel (480 g., LPS-l) eluting with hexane:
ethyl acetate (8:2) to yield 1.03 g. of (S)-[3-
methyl-1-[[3-[(phenylmethoxy)methyl~-2-(phenyl-
thio)-3H-imidazol-4-yl]carbonyl]butyl]carbamic
acid, i,1-dimethylethyl ester as a yellow solid;
m.p. 86-89. TLC (silica gel; hexane:ethyl
acetate, 1:1) Rf = 0.73.
~S Anal. calc'd. for C28H35N3O4S:
C, 65.99; H, 6.92; N, 8.24; S, 6.29
Found: C, 66.19; H, 7.13; N, 8.30; S, 6.45.
d) (S)-[3-Methyl-1-[[3-[(~henyl-e-h-oxy)methyl~ -
3H-imidazol-4-yl]carbonyl Lb~ arbamic acid,
1,1-dimethylethyl ester
The product from part (c) (0.61 g.,
1.02 mmole) is dissolved in methanol (10 ml.)
1 3 1 0 7, 2
HA375a
-95-
and hydrogenated at atmospheric pressure overnight
using 20% palladium hydroxide on carbon (0.5 g.)
as cata].yst. The reaction mixture is filtered
(Celite), recharged with 0.5 g. of catalyst, and
hydrogenated an additional 12 hours. It is then
filtered (Celite) and chromatographed on silica
gel (60 g., LPS-1) eluting with ethyl acetate:
hexane (2:3). The product containing fractions
are concentrated ln vacuo to give 0.38 g. of
(S)-[3-methyl-1-[[3-~(phenylmethoxy)methyl]-3H-
imidazol-4-yl]carbonyl]butyl]carbamic acid, -.-
l,l-dimethylethyl ester. TLC (silica gel; ethyl
acetate:hexane, 1:1) Rf = 0.45.
e) (S)~[3-Methyl-1-[~3-[(phenylmethoxy~methYl]-
3H-imidazol-4-yl]hYdroxymethvl]butyl]carbamic
acid, l,l-dimethylethYl ester
A 1 M solution of lithium triethylborohydride
(1.6 ml., 1.6 mmole) in tetrahydrofuran is added
dropwise, at 0 undr argon, to a solution of the
product from part (d) (0.16 g., 0.4 mmole) in tetra-
hydxofuran (4 ml.). After 30 minutes, O.S M hydro-
chloric acid (2 ml.) is added dropwise at 0, the
mixture is diluted with ether (200 ml.) and then
washed with water and brine, dried (MgSO4),
filtered, and concentrated ln vacuo to give 0.21 g.
of crude product. This material is stirred
overnight in 50 ml. of methanol:chloroform (1:1)
with 3 g. of Merck silica, filtered, and concentra-
ted ln vacuo~ The crude product is chromatographed
through silica gel (LPS-1, 100 g.) using a S%
methanol:chloroform solvent system. The product
t 3 ~ 0792
HA375a
-96-
containing fractions are combined and evaporated
to give 0.16 g. of (S)-[3-methyl-1-[[3-[(phenyl-
methoxy)methyl]-3H-imidazol-4-yl~hydroxymethyl]-
butyl]carbamic acid, l,1-dimethylethyl ester as a
viscous oil. TLC (silica gel; methanol:chloroform,
1:20) Rf = 0.22.
Anal. calc'd. for C22H23N3O4 H2O
C, 62.70; H, 8.04; N, 9.99
Found: C, 62.70; H, 8.49; N, 9.97.
f) ~-[(S)-l-Amino-3-methylbutyl]~3-L(phenyl-
methoxy)methyl]-3H-imidazole-4-methanol
A solution of the product from part (e)
(0.11 g., 0.26 mmole) in dichloromethane (2 ml.)
and trifluoroacetic acid (2 ml.) is stirred at
lS 10 for 30 minutes and at room temperature for 10
minutes. The reaction mixture is evaporated ln
vacuo and then concentrated from acetonitrile
(three times) to give a-[(S)-l-amino-3-methyl-
butyl]-3-[(phenylmethoxy)methyl]-3H-imidazole-
4-methanol which contains 2.5 M of trifluoro-
acetic acid.
Anal. calc'd. for C28H3SN3O4S
C, 44.90; ~, 4.71; N, 7.14
Found: C, 44.50; H, 4.80; N, 7.04
~S g) N-t(S)-l-rHvdroxyr3-[(~henylmethoxy~methYll-
3~-imidazol-4-yl]methylJ-3-methylbutyl]-N2-[N-
r(i,l-dimethyleth~y)carbonyl]-L-phenylalanyl
N -[(phenylmethoxy)methyl]-L-histidinamide
Diisopropylethylamine (0.11 ml., 0.52 mmole)
is added to 3 ml. tetrahydrofuran solution of the
crude 2.5 M trifluoroacetic acid salt product from
1 3 1 07q2
HA375a
-97-
part (f) (0.26 mmole), at 0 undr argon, followed
by the addition of N-[N-[(l,l-dimethylethoxy)-
carbonyl]-L-phenylalanyl]-l'-[(phenylmethoxy)methyl]-
L-histidine (135.9 mg., 0.26 mmole), 1-hydroxybenzo-
S triazole hydrate (135.2 mg., 0.26 mmole) and
finally dicyclohexylcarbodiimide (53.7 mg.,
0.26 mmole). After one hour, dimethylaminopyridine
(15 mg., 0.12 mmole) is added. ~he reaction
mixture is stirred overnight at room temperature,
concentrated ln vacuo, dissolved in e~hyl acetat~ .
(150 ml.), and filtered. ~he filtrate is washed
with half saturated sodium bicarbonate solution
and brine, dried (MgS04), filtered, and concentra-
ted i vacuo to give 0.17 g. of crude product. Two
flash chromatographies, one on silica gel (Merck,
80 g.) and the other on silica gel (LPS-l, 80 g.
employing the solvent system chloroform:methanol:
ammonia hydroxide (100:10:0.2) yields 60 mg. of
N-[(S)-l-[hydroxy[3-[~phenylmethoxy)methyl]-3H-
imidazol-4-yl]methyl]-3-methylbutyl]-N2~[N-[(1,1-
dimethylethoxy)carbonyl]-L-phenylalanyl]-N3-
[(phenylmethoxy)methyl]-L-histidinamide (3:7
isomer ratio).
h) N-~(S)-1-[Hydroxy(lH-imidazol-4-~l)methyl]-
3-methylbutyl]-~ ~[N-[(l,1-dimethylethoxy)carbonyll-
L-~henylalanyll L-histidinamide, 0.3 acetate salt
The product from part (g) (60 mg.,
0~74 mmole) is dissolved in methanol t4 ml.),
water (0.5 ml.), and lN hydrochloric acid (0.15 ml.)
and hydrogenated overnight using 20% palladium
hydroxide on carbon catalyst (40 mg.). The
~ 3 ~ 07q2
HA375a
-98-
reaction mixture is filtered through Celite,
concentrated in vacuo, and chromatographed on
silica (Merck, 30 g.) eluting with chloroform:
methanol:water:acetic acid ~90:20:2.5:1). The
product containing fractions are combined and
evaported to yield 27.5 mg. of product. A portion
of this sample is dissolved in ethyl acetate and
stirred with water. The organic phase is
concentrated to yield 12.1 mg. of N [(S)-1-[hydroxy-
10 (lH-imidazol-4-yl)methyl]-3-methylbutyl]-N2_[N-[(1~
dimethylethoxy)carbonyl]-L-phenylalanyl]-L-histidinamide,
0.3 acetate salt having an isomer ratio of about
1:4; m.p. 113 - 128. TLC (silica gel; chloroform:
methanol:water:acetic acid, 90:20:2.5:1) Rf = 0.19.
Anal- calc'd- for C~9H41N75 ' 0 3 C2~42
C, 60.70; H, 7.26; N, 16.74
Found: C, 60.80; H, 7.67; N, 16.68.
Example 19
(lS,2R)-N2-[N-~N2-(Cyclobutylcarbonyl~-L-lysyl]-
L-phenylalanvll-N-[l-(cyclohexylmethyl)-2-hydroxy-
2-(lH-imidazoI-2-yl)ethyll-L-histidlnamide, 3,3-
hvdrochloride
a) (lS,2R~-N2-~N-LN2-(Cyclobutylcarbonyl)-N6-
[(phenylmethoxy)carbonyl~-L-lysYl~-L-~henYlalanyll-
N-[l (cyclohexylmethyl)-2-hydroxY~2-~1-[(phenyl-
methoxy)methyl~-lH-imidazol-2-vllethyl]-N3-[(phenyl-
methoxv)methyl]-L-histidinamide
N-Methylmorpholine (586 mg., 5.79 mmole) and
dicyclohexylcarbodiimide (398 mg., 1.93 mmole) are
added to a mixture of N-[~cyclobutyl)carbonyl]-N6-
[~phenylmethoxy)carbonyl]-L-lysine (700 mg.,
1 3 1 07~2
HA375a
_99_
1.93 mmole), N2-(L-phenylalanyl)-N-[(lS,2R)-2-
cyclohexyl-l-[hydroxy[l-[(phenylmethoxy)methyl]-lH-
imida~ol-~-yl]methyl]ethyl]-N3-[(phenylmethoxy)methyl]-
L-histidinamide, trihydrochloride salt (1.72 g.,
1.~3 mmole) [prepared as described in Example 13(a)],
and l-hydroxybenzotriazole hydrate (325 mg.,
2~12 mmole) in tetrahydrofuran (20 ml.) at 0~
under argon. The reaction is kept stoppered in a
refrigerator for 3 days, then iltered, diluted
with ethyl acetate, rinsed with three 15 ml.
portions of water, 15 ml. of saturated sodium .-
bicarbonate solution, and brine, dried (MgS04)
and concentrated ln vacuo to yield 2.12 g. of
crude product. Recrystallization from methanol/
ethyl acetate gives 1.2 g. of (lS,2R)-N2-[N-[N2-
(cyclobutylcarbonyl)-N6-~(phenylmethoxy)carbonyl]-
L-lysyl]-L-phenylalanyl]-N-[l-(cyclohexylmethyl)-
2-hydroxy-2-[1-[~phenylmethoxy)methyl]-lH-imidazol-
2-yl]ethyl]-N3-[(phenylmethoxy)methyl]-L-histidinamide;
m.p. 146 - 162 (dec.); [~]D = -14-5 (c = 0.5,
methanol).
b) (lS,2R)~N2-[N-[N-_-(CYclobutylcarbonyl)-L-lysyl
L-~henylalanyl]-N- r 1-(cyclohexvlmethYl~-2-hydroxy-2-
(lH-imidazol-2-yl)ethyl]-L-histidinamide~ 3,3-
~5 hvdrochloride
A solution containing the product from part
(a) (1.05 g., 0.952 mmole), hydrazine hydrate
(476 mg., 9.52 mmole), and 20% palladium hydroxide
on carbon catalyst (300 mg.) in methanol (50 ml.)
is stirred under hydrogen for 24 hours. The
reaction mixture is then filtered, concentrated to
1 3 1 0792
HA375a
-100-
remove the methanol, and redissolved in water
~25 ml.) containing lN a~ueous hydrochloric acid
~2.63 ml.). Lyophillization gives 756 mg. of
(lS,2R)-N2-[N-[N2-(cyclobutylcarbonyl)-L-lysyl]-
L-phenylalanyl]-N-[1-(cyclohexylmethyl)-2-hydroxy-
2-(lH-imidazol-2-yl)ethyl]~L-histidinamide, 3.3
hydrochloride; m.p. 174 -187; [~]D = ~30 0
(c - 0.5, methanol). TLC (silica gel; n-butanol:
pyridine:acetic acid:water, 4:1:1:1) Rf = 0.48.
Anal- calc'd- for C38H55N95 3-3 ~Cl 3-3 H2O:
C, 50.84; H, 7.29; N, 14.04; Cl, 13.03
Found: C, 50.84; H, 7.43; N, 14.20; Cl, 12.96.
Example 20
(lS,2~)-N-[l-~Cyclohexylmeth~ hydroxy-2-(lH-
imidazol-2-yl)ethyll-N2-[1-oxo-3-phenyl-2-(phenyl~
methyl)propyl~-L-histidinamide, 2.2-trifluoroa etate
salt
a) 2,2-Bis~phenvlmethvl)propanedioic acid,
diethvl ester
Diethyl benzylmalonate (8.6 ml., 50 mmole)
is added dropwise over 5 minutes to a suspension of
sodium hydride (2.0 g., 50 mmole, of 60% dispersion
in mineral oil) in tetrahydrofuran (100 ml.). Gas
evolution is observed. When the addition is
completed, the mixture is heated at reflux for
10 minutes and is then cooled to 25. A solution
of benzyl bromide (6.5 ml., 55 mmole) in tetra-
hydrofuran (10 ml.) is added dropwise over 10
minutes, after which the mixture is stirred for 20
hours at ?5 under argon. lN Hydrochloric acid
(100 ml., 100 mmole) is then added and the mixture
1 3 1 07q2
HA375~
-101-
is extracted with e~hyl acetate. Th~ extract is
washed with saturated aqueous sodium bicarbonate
solution, dried (MgSO4), and concentrated ln
vacuo. The residue (17.8 g.) is chromatographed
on silica gel (Merck) eluting with hexane:ethyl
acetate (5:1) to give 2,2-bis(phenylmethyl)pro-
panedioic acid, diethyl ester.
b) ~-(Phenylmethyl)benzenepropanoic acid
A solution of the diethyl ester product from
part (a) (14.4 g., 4~.5 mmole) in ethanol (lO0 ml.)
and 1.0 N aqueous sodium hydroxide solution (95 ml~;
95 mmole) is heated at reflux for 48 hours. The
ethanol is removed ln vacuo and ~he remaining
aqueous mixture is acidified by the addition of lN
lS hydrochloric acid. The mixture is saturated with
sodium chloride and extracted with ethyl acetate.
The extract is dried (MgSO4) and concentrated ln
vacuo to give 9.6 g. of the dicarboxylic acid
which crystallizes on standing.
A solution of the above material (9.6 g.) in
dioxane (100 ml.~ containing concentrated
hydrochloric acid (1 ml.) is heated at reflux for
24 hours, after which it is concentrated to
dryness. The residue is crystallized from ethyl
acetate:hexane to give i.o2 g. of a-(phenylmethyl)-
benzenepropanoic acid; m.p. 86 - 87.
c) (lS,2R)-N-[l-(CyclohexylmethYl?-2-hydroxy-2-
rl-~(phenylmethoxy)methvl]-lH-imidazol-2-yllethyl]
N2-~l-oxo-3-phenvl-2-(phenYlm thyl)~ropyl]~N3-Ll-
(phenylmethoxy)methyl]-L-histidinamide
1 3 1 0792
HA375a
-102-
Triethylamine (0.53 ml., 3.8 mmole) and
dicyclohexylcarbodiimide (283 mg., 1.4 mmole) are
added to a solution of N-[(lS,2R)-l~(cyclohexyl-
methyl)-2-hydroxy-2-[l-[(phenylmethoxy)methyl]-1~-
imidazol-2-yl]ethyl] N3-~(phenylmethoxy)methyl]-L-
histidinamide (920 mg., 1.25 mmole) [prepared as
described in Example ll(e)], l-hydroxybenzotriazole
hydrate (210 mg., 1.34 mmole), and the product
from part (b) (330 mg., 1.4 mmole) in
tetrahydrofuran (5 ml.) at 0. The resulting
mixture is stirred for 18 hours at 25, after
which it is filtered. The filtrate is diluted
with ethyl acetate, washed with saturated sodium
bicarbonate solution and brine, dried, and con-
centrated. The residue is chromatographed on
silica gel (Merck) eluting with ethyl acetate:
pyridine:acetic acid:water (100:20:6:11) to give
900 mg. of (lS,2R)-N-[1-(cyclohexylmethyl)-2-
hydroxy~2-[1-[(phenylmethoxy)methyl]-lH-
imidazol-2-yl]ethyl]-N2-[1-oxo-3-phenyl-2-
(phenylmethyl)propyl]-N3-[(phenylmethoxy)-
methyl]-L-histidinamide as the major product;
[~]D = ~5 3~ (c = 1, methanol).
d) (lS,2R)-N-[l-~Cyclohexylmethyl)-2-hydroxy-2-
(l~-imidazol-2-vl)ethYl~ [1-oxo-3-phenyl-2-
(Phenylmethyl)Propyl]-L-histidinamide, 2,2
trifluoroacetate salt
A mixture of the product from part (c) (900
mg., l.l mmole), 20% palladium hydroxide on carbon
catalyst (200 mg.), and 1.0 N hydrochloric acid
(2~4 ml., 2.4 mmole) in methanol (20 ml.) is
1 3 1 07 ,2
HA375a
-103-
hydrogenated under a slow stream of hydrogen for
19 hours. The mixture is then filtered and
concentrated to dryness. The residue is dissolved
in water, activated carbon is added, and the
mixture is then millipore filtered and
lyophillized to give 590 mg. of a pinkish solid.
This solid is further purified by preparative
HPLC (YMC s 15 ODS column, 20 x 500 mm., 67%
aqueous methanol containing 1% trifluoroacetic
acid, 25 ml/min., W 220 nm. monitoring).
Fractions containing the major component
(retention time 32 minutas) are combined and
concentrated. The residue is dissolved in water
and lyophillized to give 465 mg. of (lS,2R)-N
[1-(cyclohexylmethyl)-2-hydroxy-2-(lH-imidazol-2-
yl)ethyl]-N -[l-oxo-3-phenyl-2-(phenylme~hyl)propyl~-
L-histidinamide, 2.2 trifluoroacetate salt as a
fluffy white solid; m.p. (70) 78 - 108; [~]D =
-28.0 (c = 0.70, methanol). TLC (silica gel,
ethyl acetate:pyridine:acetic acid:water,
40:20:6:11) Rf = 0.36.
Anal. alc d- f r 34 42 6 3 2 3 2
0.8 ~2
C, 54.39; H, 5.4 ; N, 9.91; F, 14.79
Found: C, 54.34; H, 5.55; N, 10.00, F, 14.73.
Example 21
(lS,2R)-N-~l-(Cyclohexylmethyl)-2-hydroxy-2-(lH-
imidazol-2-yl)ethyl]-N2-~1-oxo-3-(1-naphthaleny~
2-(1-na~hthalenylme~hyl~propyll-L-histidinamide,
d hydrochloride
a) 2,2-Bis(l~naphthalenylmethyl~propanedioic
acid, diethYl ester
Diethylmalonate (15.2 ml., 100 mmole)
is added to a suspension of sodium hydride
1 3 1 07~2
HA375a
-104-
(20Q mmole) in tetrahydrofuran (400 ml.). The
addition is accompanied by gas ev~lution. When
the addition is completed, ~he mixture is warmed
to reflux temperature to yield a homogeneous
solution. 1-Chloromethyl naphthalene ~35.4 g.,
200 mmole) is added to the refluxing solution over
one hour. THe mixture is then stirred at reflux
for 17 hours, af~er which it is ~uenched by the
addition of excess lN hydrochloric acid. The
mixture is extracted with ether and the extract is
washed with saturated sodium bicarbonate solution,_
dried (MgSO4), and concentrated to a yellow oil.
The residue is dissolved in hot petroleum ether
and the colorless solution is decanted from a
brown insoluble gum. The solu~ion is concentrated
and the residue is crystallized twice from
methanol to give 13g. of 2,2-bis(1-naphthalenyl-
methyl)propanedioic acid, diethyl ester as
colorless crystals; m. p. 79 - 81.
b) -(l-NaDhthalenylmethyl)-l-na~hthalenyl-
~ro~anoic acid
A mixture o~ the diethyl ester from part (a)
(11.0 g., 25 mmole) and 1.0 N sodium hydroxide
solution (50 ml., 50 mmole) in ethanol (50 ml.) is
stirred at 25 for 3 days. Sodium hydroxide solid
(~ g.) is then added and the mixture is stirred
for an additional 6 hours at 25~, after which it
is concentrated ln vacuo. The residue is diluted
with water, resulting in a dense white
precipitate. The precipitate is collectèd and
washed with hexane. The filtrate is then washed
1 3~ 07q2
HA375a
-105-
with hexane and the combine~ hexane washes are
extracted with sodium hydroxide Colution. The
scdium hydroxide extracts and the solid
precipitates are combined and acidified by the
addition of concentrated hydrochloric acid. The
mixture is then extracted with ethyl acetate and
the extract is dried and concentrated. The
residue (7.2 g.) is dissolved in dioxane
(250 ml.). Concentrated hydrochloric acid (1.0
ml.) is added and the mixture is stirred at 90
for l9 hours, after which it is concentrated ln '
vacuo. The residue is triturated with methanol to
give 4.6 g. of ~ naphthalenylmethyl)-1-naphtha-
lenylpropanoic acid as a white powder; m.p.
lS 168 ~ 170.
c) N-~(lS,2R)-1-(CyclohexYlmethy~-2-hydroxy-2-
(lH-imidazol-2-yl~ethyll-L-histidinamide
A mixture of [(l,l-dimethylethoxy)carbonyl]-
N-[(lS,2R)-l-(cyclohexylmethyl)-2-hydroxy 2-[1-
[(phenylmethoxy)methyl]-1~-imidazol-2-yl]ethyl]-N3-
[(phenylmethoxy)methyl]-L-histidinamide (1.46 g.,
2.0 mmole) [prepared as set forth in Example 11 (d)],
20% palladium hydroxide on carbon catalyst (350 mg.),
and 1.0 N hydrochloric acid (4 ml., 4 mmole)
in methanol (15 ml.~ is hydrogenated under a
slow stream of hydrogen for 20 hours at 25,
after which it is filtered and concentrated
to dryness. The residue (1.1 g.) is dissolved in
acetic acid l30 ml.) and dry HCl gas is bub~led
through the solution for 30 minutes at 25. The
mixture is then stirred at 25 for 2 hours, after
which is is concentrated 1n vacuo. The residue is
1 3 1 07q2
HA375a
-106-
triturated with acetonitrile to give 833 mg. of a
white solid. The solid is dissolved in excess lN
hydrochloric acid and concentrated to dryness.
The process is repeated three times to give
N-[(lS,2R)-1-(cyclohexylmethyl)~2-hydroxy 2-~lH-
imidazol~2-yl)ethyl]-L-histidinamide as a white
solid.
d) (lS,2R)-N-~-(Cvclohexylmethyl~-2-hydrox~-2-
(lH-imidazol-2-yl)ethyll-N2-[1-oxo~3-(1-naphthal-
en 1)-2-(1-na~hthalenylmethyl)pr~pyl]-L-histidinamide,
dihydrochloride
Triethylamine (O.68 ml., 4.9 mmole) and
dicyclohexylcarbodiimide (309 mg., 1.5 mmole) are
added to a mixture of the product from part (c)
(750 mg., 1.5 mmole), 1-hydroxybenzotriazole
hydrate (230 mg., 1.5 mmole) and the product from
part (b) (511 mg., 1.5 mmole) in dimethylformamide
(7 ml.) at 0. The resulting mixture is stirred
for 18 hours at 25 after which it is concentrated
to dryness. Methanol (9 ml.) and 1.0 N hydro-
chloric acid (6 ml.) are added to the residue.
The mixture is filtered and the filtrate is
concentrated to dryness. The residue is flash
chromatographed on silica gel (Merck) eluting with
ethyl acetate:pyridine:acetic acid:water
(50:20:6:11) to give a major product (Rf = 0.25)
that is homogeneous but dark brown in color. This
material is dissolved in l.0 N hydrochloric acid
and reconcentrated. The resulting hydrochloride
30 salt (430 mg.) is purified by chromatography on `~
HP-20 eluting with a gradient from 0.01 N hydro-
1 3 1 07q2
E~375a
-107-
chloric acid to methanol. Colorless fractions
containing the major product are combined and
partially concentrated to remove the methanol. The
aqueous residue is lyophillized to give a fluffy
electrostatic white solid that is dissolved in
water and relyophillized to give 220 mg. of
(lS,2R)-N-[1-(cyclohexylmethyl)-2-hydroxy-2
(lH-imidazol-2-yl)ethyl]-N2-[1-oxo-3-(1-naphthalenyl)-
2-(1-naphthalenylmethyl)propyl]-L-histidinamide,
dihydrochloride; m.p. 163 - 183; [~]D = -56.1
(c = 0.59, methanol). TLC (silica gel; ethyl
acetate:pyridine:acetic acid:water, 50:20:6:11)
Rf = 0.25.
Anal. calc d- for C42X46 6 3 2
C, 63.06; H, 6.59; N, 10.51; Cl, 9.75
Found: C, 63.03; ~, 6.76; N, 10.61; Cl, 9.78.
~xamples 22 - 46
Following the procedures of Examples 1 to
21, additional compounds within the scope of this
invention can be prepared having the forula
R5 O R4 R3
X-~ NH -CH - C ~ NH - CH- C- N~- CH- CH - R
OH
wherein the substituents are as defined below.
1 3 1 07q2
_ 108- HA375a ~`
z~
~, Q
Q Q Q
z-~ z-~
N N
~ I Q Q
o = ~ o _~
~ y ~ ~ ~ Q
o = V 2: Z
~n O O=y 0= ~
1~ ¦ ` N
~ 3 ~ 07~
HA37 5a
--109-
2~ 2
N N
~ N
U V ~
~ ¦ U U
2 [~
N y U
O=c,~ O=U ~=
~_ N I ~_ U -- U '~
O = ~ Z ~ Z
_ =~t O--y 0=~
Z U
2 2
O , . ~.
. U~ `D ' ~ ,
1 3 1 0792
HA37 5a
-- 110--
'o~ ~z =-Z~
~ ~ N
- Q
N N ~
r~ I
Z_-- O ~
y ~ _N ¦ O _ O
$ o=$ o=$ ~
- ~ o ô~ ~
r ' ,~ ~
1 3 1 0.7~2
HA37 5a
0~ z~
N
U ~ ~
~, ~, 3
~ r~ I U N ~
Z~ _ Z~
O=
O = t.~ l
<~3_ I ~N ~ 0 -D
O= Z O=y Z
n_ ~ O Z~ ~
", ~ . Y
~3
~ .
1 3 1 07q2
HA375a
112-
Q ~ ~
~ ~1 u-~
.. ,, I ~ ~ Z = Z
N N N
U U U
U UN N
~ _ 2N~2
~ I U U U
O =U
U ~ :: _ y O _ U
N I N
t I ~ [~
~ I `,
1 3 ~ 07Q2
HA37 5a
--113--
u ~ ~ ~ ~u
~`I ~ N
2~ U Q~
U U~ U
~, Z~ Z-~ z
N ~`I ~ ~ /
~ I U UU C.)
O 0=0 ~ ~ =~ ~
1 3 ~ O/7q2
_ 114- ~1A375a
=z~ ~z ~ Z~Z
? ? S'
N
O =
O = ~ ~N~
o _ ~, Cl ~:
~ N Z O -- C~ ~
,~ ~ ZO = O O ~ Z
3 ~
1 3 ~ 07q2
~A375a
-115--
~a~
1000 tabl~t~ ~ac~ corltaining th~ followi~g
in~redient~ s
N2~ lN~ Dim~thyl~thoacy)
S casbonyl ] -L-ph~nyl al 3~yl ~ N~
L ( lS ) -2 ~cyclohexyl-l- [ ( R ) -hydro~y~
l~-i;nidazol-2 -ylm~t:hy~ ~ ~thy3 ] -
L-histidina~id~, monoacot~ts
salt 250 mg.
Cornstarch lOQ mg.
G~lati~ 20 m~.
*~vicol (micxoc:~stalli~o c~llulosel~)50 mg.
~agne~ium stoa~t~
~5 D35~-
15 ar~- praparod ~OD~ ~u~ici~nt bullc gu~ti~ y
mi~cing th~ active ~nono~c~t~t~ ~lt cO~pOU~la ~d
corn~arch w$t~ a~ agu~ouc ~ utlon o~
g~latin. Th~ mixtur~ dl ~d ~o~ ~o
~n3 powd~r. q~ ~Avic~ d th~ t~ ag~
20 st~arat~ ar~ ad~s~d w~th gr~ul~o~O T~L8
mixtus~ i~ th~ comp~e~8~ iDI a t~lbl9'e ~p~081~ to
for~ 1000 t~ ~ co~ 25~ ~O o~
activ~
In a 3iln~ oa~er, t~l~t~ co~i~ q
25 2S0 mg. o~ ~ produc~ o~ ~y o~ E:x~splo~ 1 ~o 6
8 ~o 46 ca~ b~ prop&~a~.
_` A ~in~l~ proce~du~o c~ b~ ~ploy~a to ~on~
t~l6t~ co.sl~ai~ 500 laq. o~ ~c~iv~ adi~
* Trademark
.
1 3 1 ~7q2
HA375a
-116-
Exam~le 4~
An injectable solution is prepared as
follows:
N -[N-~(1,1-Dimethylethoxy)
5 carbonyl]-L-phenylalanyl]-N-
[(S)~1-[(R)-hydxoxy(lH~imidazol-
2-yl)methyl]-3-methylbutyl]-L-
histidinamide, acetate salt1000 g.
Methyl paraben 5 g.
10 Propyl paraben 1 g.
Sodium chloride 5 g. -.-
The active substance, preservatives, and
sodium chloride are dissolved in 3 liters of water
for injection and then the volume is brought up to
5 liters. The solution is filtered through a
sterile filter and aseptically filled into pre-
sterilized vials which are closed with
presteriliæed rubber closures. Each vial contains
5 ml. of solution in a concentration of 200 mg. of
active ingredient per ml. of solution for
in] ectlon.
In a similar manner, an injectable solution
containing 200 mg. of active ingredient per ml. of
solution can be prepared for the product of any of
Examples 1, 2, and 4 to 46.
1 3 1 0792
HA375a
-117-
ExamDle 49
-
lO00 tablets each containlng the following
ingredients:
N2-[N-[(lll-Dimethyle~hoxy)-
S carbonyl]-L-phenylalanyl]-N-
[ ( lS ) -2-cyclohexyl-1-[(R)~hydroxy
lH-imidazol-2-ylmethyl]ethyl]-
L-histidinamide, monoacetate
salt 500 mg.
lO Avicel 300 mg.
Hydrochlorothia7ide 14.5 mg.
Lactose 113 mg.
Cornstarch 15.5 mg.
Stearic acid 7 m~.
950 mg.
are prepared from sufficient bulk quantities
by slugging the N2-[N-[(l,1-dimethylethoxy)-
carbonyl]-L-phenylalanyl]-N-[(lS)-2-cyclohexyl-
l-[(R)-hydroxy-lH-imidazol 2-ylmethyl]ethyl]-L-
histidinamide, monoacetate salt, Avicel, and a
portion of the stearic acid. The slugs are ground
and passed through a #2 screen, then mixed with
the hydrochlorothiazide, lactose, cornstarch, and
remainder of the stearic acid. The mixture ls
compressed into 950 mg. capsule shaped tablets in
a tablet press. The tablets are scored for
dividing in half.
In a similar manner, tablets can be prepared
containing 500 mg. of the product of any of
Examples l to 6 and 8 to 46.
