Note: Descriptions are shown in the official language in which they were submitted.
~A25g~
--1--
ACYLALKYLAMINOCARBONYL SUBSTITUTED AMINO
AND IMINO ACID COMPOUNr~S
This invention is directed to the variousnovel acylalkylaminocarbcnyl substituted amino and
imino acid compounds of formula I below,
intermediates for preparing such compounds, and
compositions and methods of using compositions
containing these novel compounds.
The novel acylalkylaminocarbonyl substituted
amino and imino acids, esters, and salts of this
invention are of the formula
(I)
O ~RI 1l
R3-fH-Co(CH2)n~N--C-X
NH
C=O
R2
X is an amino or imino acid or ester of the
formula
R7
H2C fH2 H ~ ~ Rg~ ~ CH~
-N - C-COOR6 , -N - C COOR6 , -N C-COOR
~ (L) l (L) l (L)
. :
- .
.
. ~
~LZ6~
' HA25~
--2--
5>< ~ ~ R 11>~ ~ R '
-N ---C-COOR6 -N--C-COOR6 -N--C-CO~
I (L) I (L) I (L)
H H H
~)~ OR ' -N--C-COOR6 ~3
C-COOR6 , -N~
2 5 1 ( L ) H ( L )
R124
-N--CH-COOR or _N--C-COOR
6 1 (L) 6
R4 R5 H
;. . .. .
. .
~. .. .
.
.' ' ': ' '
lZ~
HA259
--3~
R7 is hydrogen, lower alkyl, halogen, keto,
O Rl~
hydroxy, -NH-C~lower alkyl, azido, amino, -~<
o R20
5 -NH-C (CH2)m- ~ , -(CH2)m - ~ (Rl 3)p
- ( CH2 ) m{~,~ ~ ~ ( CH2 ) m ~S3 , ( CH2 ) m~
a 1- or 2~naphthyl of the formula
-(CH2)m-__~%~ ~ , -(CH2)m-cycloalkyl,
O Rls (Rl4 )p
-O-C-N , -O-lower alkyl, -O-(CH2)m -
R15 (R13)p
a l- or 2-naphthyloxy of the formula
-O-(CH2)m ~ , -S-lower alkyl,
20 -S-(CH2) ~ , or a 1- or 2-naphthylthio
(Rl 3 )p
,
:~:, , . - ,
-
H~2 'j ~i a
of the formula
-S~(CH2)m ~ ( 14)p
R8 is keto, halogen, ~l5
-O-(CH2)m ~ , -O-lower alkyl, a 1- or
~R13)p
2-naphthyloxy of the formula
-O-(CH2)m ~ ( 14)p
-S-lower alkyl, -S-(CII2)m ~ (R13)p
or a l- or 2-naphthylthio of the formula
-S- (CH2);~
,J~ ( R 14 ) p
Rg is keto ox -(CH2)m ~ (R
13 p
:` :
~ ` ,
:
:' :
.
~259a
-5-
Rlo is halogen or -Y-Rl 6 ~
Rll, R 1l, Rl2 and R'l2 are independently
selected from hydrogen and lower alkyl or R'll,
Rl2 and R'l2 are hydrogen and Rll is
(Rl4 )p
Rl 3 iS hydrogen, lower alkyl of 1 to 4
carbons, lower alkoxy of 1 to 4 carbons, lower
alkylthio of 1 to 4 carbons, chloro, bromo,
fluoro, trifluoromethyl, hydroxy, phenyl, phenoxy,
phenylthio, or phenylmethyl.
Rl 4 iS hydrogen, lower alkyl of 1 to 4
carbons, lower alkoxy of 1 to 4 carbons, lower
alkylthio of 1 to 4 carbons, chloro, bromo,
lS fluoro, trifluoromethyl, or hydroxy.
m is zero, one, two, three, or four.
p is one, two or three provided that p is
more than one only if Rl3 or Rl4 is hydrogen,
methyl, methoxy, chloro, or fluoro.
Rls is hydrogen or lower alkyl of 1 to 4
carbons.
Y is oxygen or sulfur.
Rl 6 iS lower alkyl of 1 to 4 carbons,
-(CH2)m ~ , or the Rl6 groups join to
~ (Rl33p
complete an unsubstituted 5- or 6- membered ring
.
.
. :. - . . ,
: . .. . .
: . . . `- ~ ' ~
., .
,
.
~Z69~
H~25~a
--6--
or said ring in which one or more of the
carbons has a lower alkyl o 1 to 4 car~ons
or a di(lower alkyl or 1 to 4 carbons)
substituent.
R5 is hydroyen, lower alkyl,
-(CH2) ~ , -(CH2) ~ ' ( 2)r
-(CH2) ~--OH , -(CH2)r~
OH H
-(CH2)r~ N ~ -(CH2)r-NH2~ ~(CH2)r~sTT'
(CH ) -S-lower alkyl, (CH2)r ~
. NH2
o
or (CH2)r 2
;
` 35
`: :
- - -
., .. , . . . ~ . .
.
~-'' ".,: .. :,
~Z~ 6
['~.25
--7--
r is an integer ~rom 1 to 4.
Rlg is lower alkyl, benzyl, or
phenethyl.
R20 is hydrogen, lower alkyl, benzyl
or phenethyl
R4 is hydrogen, lower alkyl,
(CH2)m ~ , -(CH2)m-cycloalkyl,
2 m~ ~ ' (CH23
~S O
~(CH2)m ~ ~ or
~ ~<~
.
, . - .
.: :
`.~ ~ ~ ' ;:'. , :
;: ~ ` :' ~ . , ,
~2q~1E6
--8--
n is one or two.
Rl is hydrogen, lower alkyl, halo su'~stit!1ted
lower alkyl,
~(CH2)m ~ ~ ( 2 1 ~ S~
( 14 p
- (CH2) ~3 t - (CH2) ~)
~~cH2)m-cycloalkyl~ (CH2)2 NH2
- (CH2) 3-NH2 , - (CH2) 4-NH2
-(CHz)-r~ ~ - OH , (CH2)
OH
( 2)r~ (CH2)r-SH, -(CH2) -OH ,
: N
H ~H
(CI-I ) -S-lower alkyl, -(CH2)r \ , or
NH2
O
Il
(CH2)r 2
R2 is ~(CH2)m~ , ~(CH2)m~3
~ : 35
: `'
~2~ 6
_g_ ~A259
-~CH2)m~ ~ , or -(CH2)m--~ -
R3 is hydrogen, lower alkyl,
- ( CH~ )m--~ , - (CH~ )m l ,~J
-(CH2 )m{~J ' (CH2 )m~J
halo substituted lower alkyl, -~CH2 )m-cycloalkyl,
(~OEI
: H
:
(CH2)r ~ ~N ~ -(CH2)r-NH2 , -(CH2)r-SH,
2 0 H
.` NH
: -(CH2 )r-S-lower alkyl, -(CH2 )r-NH-C
NH2
,
,
~: ' '', ';
lZG9~
HA259~
--10--
-(CH2)r-C-NH2 or -(CH2)r-OH.
R6 is hydroyen, lower alkyl, an alkali metal
O R2 1
ion, benzyl, benzhydryl, -CH-O-C-Rl8, -C C-O-2~ ,
Rl7 R22
-CH-(CH2-OH)2, -CH2-CH-CIH2 , -(CH2)2-N(CH3)2,
OH OH
10 - CH2~3
Rl 7 i.S hydrogen, lower alkyl, cycloalkyl, or
phenyl.
~ 18 i.S hydrogen, lower alkyl, lower alkoxy,
lS or phenyl or Rl 7 and Rl 8 taken together are
-(CH2 )2~ -(CH2 )3-~ -CH=CH-, or ~ .
R2l and R2 2 are independently selected from
20 hydrogen and lower alkyl.
R2 3 iS lower alkyl.
R2 4 iS hydrogen, lower alkyl, ~
, or ~ (Rl4)p
; 25 ~ ~
;
... . ~: .
,
` `~ '` ' '; `' `
.. . ~
~Z~9~
~259a
The term lower alkyl used in defining
various symbols refers to straight or branched
chain radicals having up to seven carbons. The
preferred lower alkyl groups are up to four
carbons with methyl and ethyl mos-t prefer~ed.
Similarly the terms lower alkoxy and lower
alkylthio refer to such lower alkyl groups
attached to an oxygen or sul~ur.
The term cycloalkyl refers to saturated
rings of 3 to 7 carbon atoms with cyclopentyl 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
trifluoromethyl, which is preferred, pentafluoro-
ethyl, 2,2,2-trichloroethyl, chloromethyl, bromo-
methyl, etc.
The symbols -(CH2)m ~ , -(CH2)m ~ 3 ,
and -(CH2)m~ represent that the alkylene
bridge is attached to an available carbon atom.
. .
.
', ,~,. ' . ,.- ~ ',
:, - , . ,'
: :
.
-
- ~ ,
~Z6~
HA259a
-12-
Alm~uist et al., "Synthesis and Biological
Activity of a Ketomethylene Analogue of a
Tripeptide Inhibitor of Angiotensin Converting
Enzyme", J.Med. Chem., 1980, 23, 1392 1398,
disclose the ketomethylene compound of the forrnula
O O O ~
C-NH-CH-C-(CH2)z-C-N COOH.
CH2 ] ~ (L)
Thls and related compounds are also dis~losed by
Almquist et al. in U. S. Patent 4,329,473.
Meyer et al., "Novel Synthesis of (S)-l-
[5-(Benzoylamino)-1,4-dioxo-6-phenylhexyl]-L-
proline and Analogues: Potent Angiotensin
Converting Enzyme Inhibitors", J. Med. Chem.,
1981, 24, 964-969, disclose the synthesis and
activity of compounds of the formula
O O
` R-cH2-cH-c-(cH2)n-c-y
NH
~'
. :. ~ , ,
, ~
, . -. : . - : ~ ~ .
:: ; , : ,
., , - - ,. . - . . ,
~Z6~ 6
HA2Sga
-13-
Gravestock et al. in European Patent
Application 45161 disclose hypotensive compounds
of the formula
Rl-Y-NR2-CHR3-X-CHR11~CHR~-Co-NR~-CR~R~ -Rl.
Mercaptoacyl derivatives of proline and
substituted prolines are known to be useful hypo-
tensive agents due to their angiotensin converting
enzyme inhibition activity. Ondetti, e-t al. in U. S.
Patent 4,105,776 disclose such compounds ~7herein
the proline ring is unsubstituted or substituted by
an alkyl or hydroxy group. Ondetti, et al. in U. S.
Patent 4,154,935 disclose such compounds wherein
the proline ring is substituted with one or more
halogens. Ondetti, et al. in U. S. Patent 4,316,906
disclose such compounds wherein the proline ring is
substituted by various ethers and thioethers.
Krapcho in U. S. Patent 4,217,359 disclose such
compounds wherein the proline ring has a carbamoyloxy
substltuent. Krapcho in U. S. Patent 4,311,697
discloses compounds wherein the proline ring has a
diether, dithioether, ketal or thioketal substituent
in the 4-position. Krapcho .in U. S. Patent 4,316,905
discloses such cornpounds wherein the proline ring
has a cycloalkyl, phenyl, or phenyl-lower alkylene
sub~tituent. ondetti, et al. in U. 5.
~':
. ~ . . . . ..
.
:' ~ ', ~ '. :
; ~, .
~ :
5ga
Patent 4,234,489 disclose ~uch compourld~ wh~xein
the proline has a keto ~ titu~nt in the 5-
position. Krapcho, et al., in U. S. Pa'cent
4,310,461 di~close 3uch, compound~ wher~in th~
5 prolin~ ha~ an imldo, amldo, or am:Lns ~ubstituent
in the 4-position. Iwao, et al. in U.K. Paten'c
Application 2,027,025 disc:lose such compound~
wherein the prolin~ has an arom~tic ~ubstituent in
the 5-position.
Mercaptoacyl derivative~ o 3,4 d~hydro-
proline are di3clo~ed aa angiot~r~sirl con~rtin
erlzy~ inhibitor~ by Ondetti in U. ~. Patent
4,129, 566. ~rcaptoacyl ~iYati~ o~
thiazolidin~carboxylic acid a2ad ~ubsti;'cut~d
15 thiazolidin~c~rbo~ylic acid ar~ di~clo~d a~
angiote~in cov@rting ~nzy~ inhibito~ by ~nd~tti
in U. S. Patont 4,192,a7~ and by Yo~l~tomo
Pharma~utical Ind. in B~lgiu~ ~at~nt 86~,532.
M~rcaptoacyl deriv~tiv~ o~ dihydgoi~aindol~
20 carbo~ylic acid~ and t~l:rahyd~oi#oquinoline
carbo~yli~: acidu ar~ dig~:lo~ by l?o~lock in U.~C. Applica~io
2,0~8,863 and ~ ~Iayash~ 2t al. ir~ U. S. Pal:ont
4, ~56, 751 .
Mo~ca~toac~l d~rivati~ o various aslino
25 acids ar~ disc:losed by Ond~t~i ~t ~1. as being
,~,
' ~ '
. , :
.
-lS- HA259a
useul hypotensive agent~ due to th~ir angioten~in
converting enzym~ inhibitio~ activity in U. S.
Patent 4,053,651.
Carboxyalkylaminocarbonyl ~ubstitut~d t~tra-
hydroisoquinolin~s and proline~ ar~ di3closed a~
possessing angioten~in co~verting enzym~
inhibition activity by Tanab2 in European Patent
application 18,549 .
The compound of ~ormula I can b~ prepare~
by coupling an acylated alkylamin~ o~ th~ formul~
(II) ~ R
R3-C~-C-(C~2)~-~
1~ N}31
C--O
z
particularly the hydroc~lorid~ salt with th~ acid
chlorid~ o~ th~ ~or~ula
(II~) el-c-x
o
i~tho pxes~co o~ N-mo~hyl morpholin~ whorsi~ R~
~ h~ d~inltion o~ X i~ a~ ~a~ily re~ov~blo
estor pro~octing group such a~ ban2yl or t-butyl.
Re~o~al of th~ R~ prot~cting grou~ ~uch as by
hydrogena~ion when R~ is b~nzyl o~ ~re~t~ont with
tri~luoroac~tic aci~.wh~ Ro i8 t-bu~yl yi~ld~ tho
products of for~ula I ~h~r~in R~ i~ hydrog~n.
..
. . . . .
.
- ~
. ~ - . .
~6~
~!/`.2
--16--
The reactant of formu].a ~.I can be prepared ~y
converting the carboxyalkylamine of the form.ula
(IV) ll
HO-C-(CH2)m-N -prot
wherein prot is a protecting group such as benzylo~y-
carbonyl, to its acid chloride and then reacting
with an oxazolone of the formula
(V)
N
R2C C-R3
O - C=O
to yield
(VI)
O R
Il 11
3 1 ( 2)m P
NH
I
C=O
R2
Removal of the pr~tecting group such as by
hydrogenation yields the reactant of formula II.
The reactant of formula II wherein Rl is
other than hydrogen can also be prepared by
reacting a ketone of the formula
- ~ .. .
`. . , '
` : , ` .
.. : `: . . :
.. ~ ` , .
~2~9~
s Z ~ J,
--17--
(VII) O
Il
R3-CH-C- (CH2) n-halo
NH
. I
C=O
R2
wherein halo is Cl or Br with a substitu-ted
amine of the formula
(VIII)
Rl -NH2
The ketone intermediate of formula VII
can be prepared by treating a ketone of the
formula
(IX) O
Il
prot-NH-CH-C-(CH2)n-halo
R3
wherein prot is a protecting yroup such as
benzyloxycarbonyl with hydrogen bromide and
acetic acid followed by reaction with the acid
halide of the formula
(X) . O
R -C-halo
in the presence of base such as sodium bicarbonate.
.
. .
:`
,
. . , ~ .- ~: . , :
.
', . :'' ~ ~ .' :
~Z~9~
1~-
The compounds of formula I can also be
obtained by reactin~ a carboxyalkyla~inocarbon~l
substituted amino or ~nino acid chloride of the for~la
(XI)
~l 1l
C tc~2)n x
wherein ~6 in the definition of X is an easil~
removable ester protecting group such as benzyl
or t~butyl with the oxazolone of formula V.
Removal of the R6 ester group yields the compounds
of formula I wherein R6 is hydrogen.
The reactants of formula XI can be obtained
by treating a substituted amine of the
formula
(XII)
O Rl
H3C-O-C-(C~2)n-NH
with the acid chloride of ormula III to
yield
(XIII)
: O R O
l 1 11
H3C-O-C-(CH2)n-N- C-X
Treatment with methanol and sodium hydro~ide, follow-
ed by oxalyl chloride yields the reactantof formula XI.
The acid chloride amino or imino acid
ester of formula III is prepared by treating the
.
~`
,:
, . . . .
-, ~ :
, ~ ,.
:. :.:
.
. .
,
,
- : . .
I-IA ~ 5 9 ,~
--19--
corresponding amino or irnino acid ester hydro-
chloride with phosgene in the presence of
N-methyl morpholine.
In the above reactions if any or all o~
Rl, R3 and R5 are
-(CH2) ~ OH , -(CH2~r ~ OH, -(Cr{2)r~0H,
OH
lG ~(CH2)r~NH2' -(CH2) ~ N , -(CH2) -SH, or
N
H
~ NH
-(CH2)r-NH-C ~
NH2
then the hydroxyl, amino, imidazolyl, mercaptan
or guanidinyl function should be protected
during the reaction. Suitable protecting 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 acld, or other known methods following
completion of the reaction.
The ester products of formula I wherein
R6 is lower alkyl, benzyl or benzhydryl can be
chemically treated such as with
. . ' ~ ' , : ` ~ `'
~ . . . : . .- . .
: '' .' ~,., :.
:: ~
. . .
121E~9~b'~
HA25~a
--~0--
sodium hydroxide in aqueous dioxane or with
trimethylsilylbromide to yield the products of
formula I wherein R6 is hydrogen. The benzyl and
benzhydryl esters can also be hydrogenated, for
example by treating with hydrogen in the presence
of a palladium on carbon catalyst.
The ester products of formula I wherein R6
l
iS -CH-O-C-Rl 8 may be obtained by employing the
Rl 7
acid chloride of formula III in the above
reactions with such ester group already in place.
Such ester reactants can be prepared by treating
the corresponding amino or imino acid of the
formula
(XIV) HX
wherein R6 is hydrogen with an acid chloride such
o O
20 ~as ~ ~ CH2~O-C-Cl or (H3C)3-C-O-C-Cl
SO dS to protect the N-atom. The protected
:
'` ~: : ~ :
- . . . . . . .
- . , .
.
~25ga
-Zl-
amino or imino acid is then reacted in the
presence of a base with a compound of the formula
(~V) o
L-CH-O-C-Rl 8
E~17
wherein L is a leaving group such as chlorine,
bromine, tolylsulfonyl, etc., followed by removal
of the N-protecting group such as by treatment
with acid or hydrogenation.
The ester products of formula I wherein R6
o
iS -CH-O-C-Rl 8 can also be obtained by treating
~17
the product with formula I wherein R6 is hydrogen
with a molar excess of the compound of formula XV.
The ester products of formula I wherein R6
2 1 1i
is -C - C-O-R23 can be prepared by treating the
20 R22
product of formula I wherein R6 is hydrogen with a
molar excess of the compound of the
~` :
. :
.'
,, . , ~'. '
. ' '~, ~, ~ ' , ' '
~' ' ' , ' ,' .
. ' ' , ' . .
.~ ' . "
`, ' ' '
,
IIAZ 5 ~
-22-
formula
(XVI)
1 21 ll
I-C-- C-O-R23
R22
The ester producks of formula I wherein
R6 is -CH-(CH2-OH)2 or -CH2-CH - CH2 can
OH OH
be prepared by coupling the product of formula I
wherein R6 is hydrogen with a molar excess of the
compound of the formula
(XVII)
CH-(CH2-OProt)2
OH
or the formula
(XVIII)
CH2- CH - ICH2
OH OProt OProt
in the presence of a coupling agent such as
: dicyclohexylcarbodiimide followed by removal
of the hydroxyl protecting groups.
Similarly, the ester products of
formula I wherein R6 is -(CH2)2-N(CH3)2 or
'
~; ~ - , ,: .:
-.-... ,
23-
-CH2 ~ can be prepared by coupling the
product of formula I wherein R6 is hydrogen
with a molar excess of the compound of the formula
(XVIX)
HO-CH2-CH2-N (CH3)2
or the formula
(XX)
HO-CH2~)
in the presence of a coupling agent such as
dicyclohexylcarbodiimide.
The esters of formula I wherein R6 is
lower alkyl can be obtained from the carboxylic
acid compounds, i.e., wherein R6 i5 hydrogen,
by conventional esterification procedures, e.g.,
treatment with an alkyl halide of the formula
R6-halo or an alcohol of the formula R6-OH.
The products of formula I wherein R7 is
amino may be obtained by reducing the corresponding
products of formula I wherein R7 is azido.
Preferred compounds of this invention
with respect to the amino or imino acid or
ester part of the structure of formula I
are those wherein:
.,
.
~ 35
.~ :
: :
, :
":
:~'
: ~ -
,, . ~
.
.
- ,
9~
f~:25~cl -
-24-
is hydrogen, cyclohexyl, or phen~
R5 is hydrogen, straiyht or branched
chain lower alkyl or 1 to 4 carbons,
-CH2 ~ , -C7-7 ~ H
-CH2 ~ OH , CH2
OHN
~ J (CH2)~ 2
N
H
~ NH
-CH2-sH~ -(CH2)2 S CH3 ~ -(CH2)3NHC
NH2
O O
il 11
-CH -C-NH2 or ( 2)2 2
.
3S
. : .
:
,
:
126~
~IA25ga
-25-
R6 ls hydrogen, straight or branched chain
lower alkyl of 1 to 4 carbons, alkali rnetal salt,
O O
-Cll-O-C-R18 , -CH2-C-OR23
R17
-CH-(cH2-OH)2 ~ -CH2-cH - CH2 , -(cH2)2~1(c~3)2
OH OH
or -CH2 ~ .
N
` ~ R23 is straighk or branched chain lower
alkyl of 1 to 4 carbons, especially -C(CH3)3.
R17 is hydrogen, straight or branched
chain lower alkyl of 1 to 4 carbons, or
cyclohexyl.
R18 is straight or branched chain lower
alkyl of 1 to 4 carbons or phenyl.
R7 is hydrogen.
R7 is hydroxy.
~ R7 is straight or branched chain lower
alkyl of 1 to 4 carbons or cyclohexyl.
.
` ,
.
~ 35~
.
.` : ~ ~ , .
i ~ ,
i; .. . . : ~ ..
- . .
~Z~
f~259a
-26-
R7 is amino.
R7 is -O-lower alkyl wherein lower alkyl is
straight or branched chain of 1 to 4 carbons.
R7 is -(CH2)m - ~ wherein m is
zero, one or two and Rl 3 iS hydrogen, methyl,
methoxy, methylthio, chloro, bromo, fluoro, or
hydroxy.
R7 is -O-(CH2)m -
Rl3
l-naphthyloxy or 2-naphthyloxy wherein m is zero,
one, or two and R1 3 iS hydrogen, methyl, methoxy,
methylthio, chloro, bromo, fluoro, or hydroxy.
R7 is -S-lower alkyl wherein lower alkyl is
straight or branched chain o 1 to 4 carbons.
R7 is -S-(CH2)m - ~ R1 3
l-naphthylthio, or 2-naphthylthio wherein m is
zero, one, or two and R13 is hydrogen, methyl,
methoxy, methylthio, chloro, bromo, fluoro, or
hydroxy.
R8 is -O-lower ~lkyl wherein lower alkyl is
straight or branched chain of 1 to 4 carbons.
. ' ' ' , ` . . ~ - . -
.: .... :. ~ .. .. ..
. . ~ .- . . , . , , . - . .
. . - . . ~ ,
, . ~.. .. , . : ,
-: . ~.' ': .
ElA259a
-27-
R8 is -O-(CH2)m ~ Rl3 wherein m is
zero, one, or two and Rl 3 iS hydrogen, methyl,
methoxy, methylthio, chloro, bromo, fluoro, or
hydroxy.
R8 is -S-lower alkyl wherein lower alkyl is
straight or branched chain of 1 to 4 carbons.
R8 is -S-(CH2)m- ~ Rl3 wherein m is
zero, one or two and Rl3 is hydrogen, methyl,
methoxy, methylthio, chloro, bromo, fluoro or
hydroxy.
- R9 is phenyl, 2 hydroxyphenyl, or 4-hydroxy-
phenyl.
Rlo are both fluoro or chloro.
Rlo are both -Y-Rl 6 wherein Y is O or S, Rl 6
is straight or branched chain lower alkyl of 1 to
4 carbons or the Rl 6 groups join to complete an
unsubstituted 5-or 6-membered ring or said ring in
which one or more of the available carbons has a
methyl or dimethyl substituent.
Rll, R'll, Rl2 and R'l2 are all hydrogen, or
Rll is phenyl, 2-hydroxyphenyl, or 4-hydroxyphenyl
and R'll, Rl2 and R'l2 are hydrogen.
R2 4 i S phenyl.
Most preferred compounds of this invention with
respect to the amino or imino acid or ester part
of the structure of formula I are those wherein:
;
:
...... .. ~ ~ , ,
.
- - .
~z~
~1~s25
-22-
X is
~ (CH2)t~
~ S ~
H2f CH2 H2 ~C CH2
-N C-COOR6 or -N ~ (CL)OO 6
O o
R6 is hy~ogen, -CH-O-C-CH3, -CH-O-C C2}!5
CH3 CH (CH3) z
O O
-cH-o-c-c2Hs ~ -CH -o-c-c2H5
~ CH3 0
O
`~ 25-CH2-0-C-C(CH3)3 , an alkali metal salt,
: : straight or branched chain lower alkyl of
: ~ l to 4 carbons, --(CH2)2N(CH3)2 or
~ CH2 ~
30 : N :
,i, :
:~ : 35
` ``. ~ ' ' : ,
HA259a
-29-
R7 is hydrogen, cyclohexyl, lower alkoxy of
1 to 4 carbons, -(CH2)m -
~0-(CH2)m- ~ , or -S-(CH2)m- ~
R~3 Rl3
wherein m is zero, one, or two and Rl 3 iS
hydrogen, methyl, methoxy, methylthio, Cl, Br, F,
or hydroxy, especially preferred wherein R7 is
hydrogen.
t is two or three, especially where t is two.
Preferred compounds of this invention with
respect to the acylalkylaminocarbonyl portion of
the structure of formula I are those wherein:
R~ is straight or branched chain lower alkyl
of 1 to 4 carbons, -CF3, -(CH2)2-NH2, -(CH2)3-NH2,
-(CH2 )4-NH2, -cH2-oH~ -CH2~> , -CH~ ~OH,
-CH2--~ OH, -CH2~ , -CH2~N
H H
~NH
CH2 -SH, - ( CH2 ) 2 -S-CH3, - ( CH2 ) 3NHC~
NH2
0 0
-CH2-C-NH2, or -(CH2) 2 -C-NH2, especially methyl.
.. , .. ., . : ,
- ~ .,
,~ . .
~L2~
HA2 5ga
-30-
R2 is -(CH2)m - ~ wherein m is
zero, one, or two and Rl~ is hydrogen, rnethyl,
methoxy, methylthio, Cl, Br, F or hydroxy,
especially phenyl.
R3 is straight or branched chain lower alkyl
of 1 to 4 carbons, -~CH2)r-NH2, -(CH2)m ~ R
1 4
-CH2 ~ ~ or -CH2 ~ wherein m is
zero, one, or two, Rl4 is hydrogen, methyl,
methoxy, methylthio, Cl, Br, F, or hydroxy, and r
is an integer from 1 to 4, especially benzyl.
The compounds of formula I wherein R6 is
hydrogen form salts with a variety of inorganic or
organic bases. The nontoxic, pharmaceutically
acceptable salts are preferred, although other
salts are also useful in isolating or purifying
,. . : ,: ~, ,
.
. ; ~
f~A~S~a
31-
the product. Such pharmaceu~ically acceptable
salts include metal salts such as sodiutn,
potassium or lithium, alkaline earth metal
salts such as calcium or magnesium, and
salts derived from amino acids such as arginine,
lysine, etc. The salts are obtained ~ reac-ting
the acid form of the compound with an
equivalent of the base supplying the desired
ion in a medium in which the salt precipitates
or in aqueous medium and then lyophilizing.
Similarly, the compounds of formula I,
especlally wherein R6 is an ester group,
form salts with a variety of inorganic and
organic acids. Again, the non-toxic
pharmaceutically acceptable salts are
preferred, although other salts are also
useful in isolating or purifying the product.
Such pharmaceutically acceptable salts
include those formed with hydrochloric acid,
methanesulfonic acid, sulfuric acid, maleic
acid, etc. The salts are obtained by
reacting the product with an equivalent
amount o the acid in a medium in which the
` salt precipitates.
-~
~::
~` 35
.`~ .
.
,: .: :
. ~.. :. :
,
.: : :
.
~lZ~9~
! ll~ 2 '; ~a
-32-
As shown above, the amino or imino acidportion of the molecule of the products of
formula I is in the L-configuration. An
asymmetric center is also present in the
acylalkylaminocarbonyl portion of the molecule
when R3 is other than hydrogen. Thus, the
compounds of formula I can exist in
diastereoisomerlc 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.
.
. . . ~ :
.: ',:' : ,. . ..
.: ~, -. .
"'~"' '',' ' ~ ' '
.
~2~ 6
~A25
-33-
The products of formula I wherein the imino
acid ring is monosubstituted give rise to cis-
trans isomerism. The configuration of the final
product will depend upon the configuration of the
R7, R8 and R9 substituent in the starting material
of formula XIV.
The compounds of formula I, and the pharma-
ceutically acceptable salts thereof, are
hypotensive agents. They inhibit the conversion
of the decapeptide angiotensin I to angiotensin II
and, therefore, are useful in reducing or
relieving angiotensin related hypertension. The
action of the enzyme renin on angiotensinogen, a
pseudoglobulin in blood, produces angiotensin I.
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
forms of hypertension in various mammalian
species, e.g., humans. The compounds of this
invention intervene in the angiotensinogen ~
(renin) ~ angiotensin I ~ angiotensin II sequence
by inhibiting angiotensin converting enzyme 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.
:
~ . -
~1~2~9
-3~-
A single dose, or preferably t~10 to four divided
daily doses, provided on a basis of about O.l to
lO0 mg., preferably about l to 50 mg., per
kilogram of body weight per day is appropriate to
reduce blood pressure. The substance i5
preferably administered orally, but paren-teral
routes such as the subcutaneous, intramuscular,
intravenous or intraperitoneal routes can also be
employed.
The compounds of this invention can also be
formulated in combination with a diuretic for the
treatment of hypertension. A combination product
comprising a compound of this invention and a
diuretic can be administered in an effective
amount which comprises a total daily dosage of
abou 30 to 600 mg., preferably about 30 to 330 mg.
of a 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, hydroflume-
thiazide, bendroflumethiazide, methychlothiazide,
trichloromethiazide, polythiazide or ben~thiazide
as well as ethacrynic acid, ticrynafen,
chlorthalidone, furosemide, musolimine,
bumetanide, triamterene, amiloride and spiro-
nolactone and salts of such compounds.
The compounds of formula I can be formulated
for use in the reduction of blood
-
: ' '. ', : .. - '~ . '
. :
.
-
. :
;gi~i
~IA2S9a
-35-
pressure in compositions such as tablets, capsules
or elixirs for oral adrninistration, or in sterile
solutions or suspensions for parenteral
administration. About 10 to 500 mg. of a compound
of formula I is compounded with physiologically
acceptable ~ehicle, 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 compositions or preparations is
such that a suitable dosage in the range indicated
is obtained.
The compounds of formula I wherein X is
-NH-CH-COOR6 also possess enkephalinase inhibition
Rs
activity and are useful as analgesic agents.
Thus, by the administration of a composition
containing one or a combination of such compounds
of formula I or a pharmaceutically acceptable salt
thereof, pain is alleviated in the mammalian
host. A single dose, or preferably two to four
divided daily doses, provided on a basis of about
0.1 to about 100 mg. per kilogram of body weight
per day, preferably about 1 to about 50 mg. per
kilogram per day, produces the desired analgesic
activity. The composition is preferably
administered orally but parenteral routes such as
subcutaneous can also be employed.
The following examples are illustrative of
the invention. Temperatures are given in degrees
centigrade.
' ' ~' ~'
.
' . ' . ,
~ 2
-36-
xample 1
(+)-1-[[[3-(Benzo~lamino)-2-oxo-4-~hen~lbutyll-
methylamino]carbonyl]-L-~roline
a) [3-(Benzoylamino)-2-oxo~ nylbutyll-
~ lcarbamic acid, phenylmethyl ester
N-methyl-N-[(phenylmethoxy)carbon~l]-
glycine (2.23 g., 10 mmole) is dissolved in
30 ml. of tetrahydrofuran and cooled in an
ice-bath. Oxalyl chloride (1 ml., 11.5 ~mole)
is added followed by 2 drops of dimethylformamide.
After stirring for 30 minutes in the ice-ba~h,
the mixture is then stirred at room temperature
for an hour. To this 0.25 ml. of oxalyl chloride
is added. The mixture is evaporated, redissolved
in 15 ml. of tetrahydrofuran, and stirred
in an ice bath. A solution of 2-phenyl-4-
(phenylmethyl)-5(4H)-oxazolone (3.1 g., 12.4 rnmole)
dissolved in 15 ml. of tetrahydrofuran is
added to the above solution stirring in the
ice-bath. Triethylamine (1.4 ml., 10 mmole) is
- added and the solution is stirred at room
temperature overnight. The precipitated
` triethylamine hydrochloride salt is filtered off.
Tetrahydrofuran is removed from the residue and
it is then redissolved in pyridine (5 ml.) and
p-dimethylamino pyridine (20 mg.) is added. After
stirring at room temperature for 3 hours, acetic
acid (5 ml.) is added and the reaction mixture is
kept at 105 for 30 minutes. The reaction mixture
is then evaporated, the residue is dissolved in ethyl
,:
:
.
` '., `', ' , . ` ,~
9~
}I~.2~
.
-37-
acetate, and washed with aqueous sodiurn bic~r~onate
and water. After trituration with eth~l acGtate/
hexane, 2.2 g. o~ homoyeneous [3-(benzoylarr,ino~-
2-oxo-4-phenylbutyl]methylcarbamic acid, p'nenyl-
methyl ester is obtained; m.p. 140-141.
b) (+)-N-[3~ ethylamino)-2-oxo-1-(phen~lmethvl)-
propyl]benzamide, hydrochloride
[3-(Benzoylamino)-2-oxo-4-phenylbutyl~methyl-
carbamic acid, phenylmethyl ester ( 0.5 g.) is
dissolved in ethanol (50 ml.) containiny lN
hydrochloric acid (2 ml.). Palladium carbon
catalyst (10~, 100 mg.) is added and hydrogenation
is continued overnight. The reaction mixture is
then filtered, evaporated, dissolved in water,
and lyophilized to 300 mg. of (+)-N-[3-(methyl-
amino)-Z-oxo-l-(phenylmethyl)propyl]benzamide,
hydrochloride as a homogeneous white powder.
c) (+)-l-[[~3-(Benzoylamino)-2-oxo-4-phen~
butyl]methylamino]carbonyl]-L-~roline, phenvl-
methyl ester
L-Proline, phenylmethyl ester, hydrochloride
(300 mg., 1.25 mmole) is dissolved in 5 ml. of
methylene chloride and N-methyl morpholine
(0.35 ml., 3.13 mmole) is added. To this solution
stirring at -20~, 12% phosgene solution in
benzene (2 ml., approximately 1.9 mmole) is
added. Stirring is continued at -20 for
30 minutes. The mixture is then evaporated,
the residue is suspended in methylene chloride
(5 ml.) and (+)-N-[3-(methylamino)-2-oxo-1-
.. . . .
,'.: .' ~: ' '
~lZ~3t;6~;
~l~2, Ji.
-38-
(phenylmethyl)propyl~benzamide, hydrochloride
(250 mg., 0.76 mmole) is added followed by
N-methyl morpholine (0.22 ml., 2 mmole). ;r~.e reaction
mixture is stirred overnight. It is then
evaporated, the residu2 is dissolved in et~yl
aceta~e and washed with saturated sodium bicarhonate,
dilute hydrochloric acid, and water. The eth~l
acetate solution after evaporation is chrornatographed
over silica gel using the solvent system, ethyl
acetate:benzene (4:6) to give (+)~ [[3-(benzoyl-
amino)-2-oxo-4-phenylbutyl]methylamino]carbonyl]-
L-proline, phenylmethyl ester (37%) as an oil.
d) (+)-1-[[[3 (Benzovlamino)-2-oxo-4-phenylbutYl]-
methylamino]carbonyl]-L-proline
The phenylmethyl ester product from part (c)
(1.0 g., 1.9 mmole) is dissolved in absolute
ethanol (75 ml.). Palladium carbon catalyst
(10%, 100 mg.) is added and hydrogenation i5
continued for 48 hours. ~he mixture is then
filtered, evaporated and chromatographed over
silica gel using the solvent system chloro~crm:
methanol:acetic acid (9.0:0.5:0.5) to give
400 mg. of (~)-1-[[[3-(benzoylamino)-2-oxo-4-
phenylbutyl]methylamino]carbonyl]-L-proline;
m.p. 75-95;Rf = 0.27 [silica gel, chloroform:
methanol:acetic acid (9.0:0.5:0.5)].
Anal. calc'd. for C24H27N3 5 2
C, 64.52; H, 6.32; N, 9.41
Found: C, 64.52; H, 6.29; N, 9.25.
..
. .
, - , . .
.- . , , ~ -
. . : :.- . ... , :
.. . ., . : .: :
:. : . :
: , :.
-''''' '' :~ ' " ', ' ', '.
,:
~z~
5 9 a
-3 9 -
Example 2
1-[[[3~(Benzoylamino)-2-oxo-4-phen~_b~~l]me ~y1-
amino]carbonyl]-L-proline, isomer A
The diastereomeric produc~ from E~.ample 1
(1 g., 2.285 mmole) is dissolved in ethyl acetate
(15 ml.). To this dicyclohexylamine (0.4g ~1.,
2.5 mmole) is added. This yields a firs~ crop
of dicycloehxylamine salt [347 mg.; m.p. 154-155;
[~]25 = _57.5o (methanol)] and a second crop
of dicyclohexylamine salt (272 mg., m.p.
153-154). The mother liquor upon evaporation
and redissolution in acetonitrile affords
another crop [110 mg., m.p. 154-155; [~]D = -48
(methnaol)]. After pooling these various crops and
upon repeated recrystallization from acetonitrile
400 mg. of pure 1-[[[3-(benzoylamino)-2-oxo-4-
` phenylbutyl]methylamino]carbonyl]-L-proline,
dicyclohexylamine salt, isomer A is obtained;
m.p. (155) 156-157; []D = -68.2 ImetAanol).
This material is suspended in ethyl acetate
and acidified with 10~ potassium bisulfate to
give 279 mg. of 1-[[[3-(benzoylamino)-2-oxo-4-
phenylbutyl]methylamino]carbonyl]-L-proline,
isomer A; m.p. 60-75; [~]D3 -86.3 (c = 1.01,
methanol). Rf 0.48 (silica gel; chloroform:
methanol:acetic acid; 90:3:3).
24 27 3 5 2
C, 69.32; H, 8.17, N, 8.98;
Found: C, 69.32; H, 8.03, N, 9.04.
'~
: :
~' ' ;'
.
:`
~9~
HA259a
~ 40-
E~.ample 3
1-[[[3-(Benzoylamino)-2-oxo-4-phenylbutt~l~meth~l-
amino]carbon~l]-L-proline, isomer B
Following the procedure of Example 2,
the mother liquor after removal of the three
crops of dicyclohexylarnine salt is con~erted
to the ~ree acid (400 mg.) and i5 chromatographed
on silica gel (chloroform:methanol:acetic acid,
90:3:3) to yield 278 mg. This material is
treated with dicyclohexylamine in acetonitrile
and recrystallized from ethyl acetate to give
80 mg. of 1 [[[3-(benzoylamino)-2-oxo-4-
phenylbutyl]methylamino]carbonyl]-L-proline,
isomer B, dicyclohexylamine salt: m.p. (139)
140; [a]D +29.3 (methanol). This dicyclo-
hexylamine salt is suspended in ethyl acetate
and acidified with 10% potassium bisulfate
to give 42 mg. of 1-[[[3-(benzoylamino)-2-oxo-4-
phenylbutyl]methylamino]carbonyl]-L-proline,
isomer B; m.p. 60-82; [a]D = +38.6 (c = 0.88,
methanol). Rf 0.48 (silica gel; chloroform.
methanol:acetic acid; 90:3:3).
Anal. calc d 24 27 3 5 2
C, 68.61; H, 8.20; N, 8.89
25 Found: C, 68.61; H, 7.98; N, 8.83.
:
.
~ 35
: ., - , : .- , - . .
- , . :
--
,.: . . : , . . . .
.: . . ;.: ' ,
6~
_41~
Examples 4 - 64
Following the procedure of Exarnple 1 the
carboxyalkylamine shown in Col. I is converted to
its acid chloride and then reacted with the
oxazolone of Col. II. Removal of the
benzyloxycarbonyl protectiny group gives the
intermediate shown in Col. III. Treatment with
the acid chloride amino or imino acid ester
of Col. IV gives the ester product shown in
Col.. V. Removal of the R6 ester grou? yields
the final product wherein R6 is hydrogen.
`~ 35
.
... .
~2~
- i~l 2 5
--42--
Col. I
HO-C- (CH2) n-N-C-O-CH2~)
0
Col. II
,~ \
2 C- R3
O ~-- C=O
Col. III
O R
11
R3-CH-C- (CH2) n-NH
2 0 ~NH
C=O
Col. IV
Il
Cl-C-X
:~:
'
: ~ :
~: :
. ~. '. :
., ~ . - ,: .... : . - , :
::, .- , . . ..
,:., ' .: '
lZ~
~11' 2 5 j ~1 "
Col. V
Il l 11
R3-CH-C- (CM2) n~M--C-X
NH
I
C=O
Rz
~0
Z5
:
:
3 5
, - ~, . . :
. ~
.. :: : ~.. , : .,
.~:: , ,, ::: -
, .
126~6
ii,., ~;, ~
s ~ ~ ~
~ ~ 8 ,~ "' o -
1~ x 1 {~
~, o
~ ~ ~
X~ I
I ~ ~ ,,
U~ Uu~
:
,
X
~: ~,,
:`; 35
.~ .
~`
.. , . . : -
.` `, . . :: .
~, -`: , ,
. . ~ . `, : : .
. ` . . . ~ . .
~2~9~;6
1'~ 2 ~, 'i i~
_a,5_
10 ~C~CI= ~), N~ ~
"~ _
2 0 : U ~, U
X~
.
~`~
2 5
3:
' U~ ~
o
:
: :
: 35
`:
~ .
-
5 (~
r1 o U
8 g
(I - ~ )- (_~^
1O x I < }c ~ ~r r ~ 7~--
~ I
a; I
~r~ J~u~
,~
~r~
~ rJ
` 30
o r~
.
.
:::
'~ :
- . . . :: ..... ..
.: . ` .: - :,-.
..... . : : :.
: -- : : : . .
` :~ - - . : ,
~Z69~ ,2 ~'3il
--47--
~ 5:
o ~ O ,.~ ¦
10 XI ~ [u~X-z
~ I
I
I
2 0 ~ (~æ (~)
: 30
:` Q)
~1
.`
: E
' . ~1
~ ` .
.~ , .
. :
.. ~ ' . .
`: :
- :
,
- ~ . . .
.: . ~ ~ -: . , . -
` :`: - . ~ : . . '
: .
~ ~ .
i~6~
. . .
--48-
~ J
U ~t`l ^
10 ~ I L'~l o=u{~- G~
~ ='o
.--J
U,~
X
: :
: ~ I ~
`~
",
~30 ~:
, , ~
:: ~ 3 5: ~
:`. : : . . .
~69til~
h2"j-l
.
:~ 15
U~
U~
,
2 5 :
~C~
O
L';
~ 30
~ '
3 5:
':
.` ~ ,~,, , : -' . . , :
, : ~ : . -
~z~ ¢
-so-
5 (~ (TO)
rJ rJ ~
r~ t I t~ tJ O
~ ~ d [~ ~ ~Z'/ ~
r~
1 r
2 0 r~
~ ¦ o I
' ~0 ~ ~
:
3 0:
:
:
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.1
~ 35
.~
. ~:
~..... : : :
,: .: , : ~ : .
~ .~ ~ .. . . .
: - ` ~ ,...... . . ..
.,, .- ` ~ -: .
~L~6~
r:"~,
--51--
~ m ~ o -- u_
10 ~ zl~
_r~.
~ I
.
,-~ 1
~ Ur, U~ ~ r
3 0 ~4 ~
~ ul ~ ~ X
~1 r,~
3 5 ~
~ ~ .
- .
:- :: - :, ., -: -
~. : : - ' -.: . - . . ,:
~Z~;9~;~ib;
~1/ , ,! '; '), _
--52--
~ ,~' 3, '~
r I ~
2 5 C
r,) r~ 2
C~
. : ~
r~d N O
, :
:: ~ : .: . . :
'-.: . . :, '~ ' . : ' ,
: .
:: : -
: . .
-
~IL;269t~
2 s
--53--
(~ O,~J
u 8 c,
~-3 ~3 c~-3
~1 ~`J
U U U
~ z (~)
20 ~ ~ u
"5
,
Z
x~ 1 u~
u~ ~" æ z
,1
: :~
: : : : 35
::
:'
: : : :
:
.. , . ` ~ . ~ ` ~ .. . . . .
, -: . .
~ ! t~
~S~--
5 (~
~ ~ I r~
~ I LX: ~, ( s
20 (~
N
~ t
~ U~
--æ (~ æ Url
..
3 5 2~ r~ r~ r
.~
.
~: .: ,,
:, - ` . : :
: - ' ,
: : ,
, -
~69~
I l, I r ~J
-55-
a ~ ~_
V ~
~; I ,~ U ,
~æ
~`
2 5
U U
~Z~ O = o
~ U~ ~
;q ~ , ~ .:r .
- 3 5
~ ::
:: :
- , - - -~ - - :
- ~ , :,. .
, ~ , ` ~.. . ,
, : ~ ` ~ . . ` :
``'` ` . ` ~ . ` ` ` ` : :
lZb`9~
-J 'j i i
--5~--
xl C~Z ~? ~n~'
~ y ,
~ -
~ O ~ ~
o~
: 25
: ,1 1
P; I
3 0:
C~
~ ~ ~ C ~ ~ .
X
: ~ IL1
35~ '
: - :~ : :
,~ . :
. . .
¢~
tL4 ~ 5 ~J~'
-57--
5 (~
~ ~ a ~ r~_
1 5 t~ ~ ~
0
:
-
:
U~
: ~, E,. ~ : .
30 ~
,
U
~ Ul
`.` ' ~ ~
~`~ 35
:
~: : :
~: `
. ~ ~
: ~ . ~ . . -
~` ~ : :, ,, .. . . . - . ,
-., ~ ,, . ~ . . .
~L2~;9~6
'J I
--58--
.
s ,~ ~ s ~z~ u f,=~z-~
10 x I
Z Z Z
~ u
2 5
~J
~C
30: ~ ~ ~
: ~ :
a~ o
0
: : 3 5
: -' :.: - .: -
~ ' ': ~ ' ",':, . '
:~2~;9~6
' '
--5r)--
d _ o (~ J \U/ (~ w
1 o X I W W ~ W
æ z
û ~ u
C
25 ~ c I _, ,,
30 ~
,
,,~
3 5
:
:: :
~` . ~ .~` . . ` ..
3~
60-
^ 8 8
~ U
U
2 5
~ I U~ U~, U~
~ :
.
: :: 35
~ : `
`:
. ` .. ., , . . ~ ~, .
. .
. : , :
: ` ~ . ~ -, ~ , .
.``,, .,. ,.: .
~ !i"s ~
-61-
U U rJ t )
o_uo--ux~ o--u
5:~ q_u ,~-~
o . o o
10 ~ [~
u y ~,
2 0
, ~ z
: ~
3 0
:: : ~
3 5
. .: ~ :. -, . -; .
. . . .
.. . ~ , ,
.
~z~
.. ~!A2,'J
--62--
.
o o=2 o=o ~
o=o ~
lo xl Czt-$ C-$ cr~
-
$
C~
,
:
~ , t .
1~
-
a)
~
:~ .
3 5
:
: :
- ,., ,~ . .. , . :
: :. ~: - ~ ,,, : . . '
~Z~`~3~
J9;1
-~3-
The Rl protecting groups in Examples 1~, 3
to 38,40 and 41, the R3 protecting groups in
Examples 42 and 43, and the R5 protectin~ yrou~
in Examples 48, 49, and 51 to 54 are remo~ed
as the last step in the synthesis. The R6 ester
groups shown in Examples 59 to 64 are not
removed.
Example 65
~ [[[4-(Benzoylamino)-3-oxo-5~E~henylpent
methylamino]carbonyl]-L-proline
a)_ 3-(Methylamino)propanoic acid, methyl es-ter
Methyl amine (66 ml.) in ethanol is
chilled with stirring in an ice-bath. Methyl
acrylate (45 ml.) is added dropwise over a
period of 20 minutes. The bath is removed after
one hour and after 4 hours the reaction mix~ure ic
concentrated ln vacuo. The liquid is distilled
at 15 mm. of Hg. at 61-63 to give 18g. of
3-(methylamino)propanoic acid, methyl ester.
b) 1-[[(3-Methoxy-3-oxopropvl)methylaminc]-
carbonyl]-L-proline, l,l-dimethylethyl ester
L-Proline, l,l-dimethylethyl ester
(8.55 g.) is taken up into 200 ml. of methylene
chloride with stirring at -20. A solution of p'~s-
gene in benzene (12.5% by weight, 60ml.)is added
followed by 8.25 ml. of N-methyl morpholine. After 30 munutes
at -20 the reaction mixture is concentrated
n vacuo. The residue is taken up into 100 ml.
of methylene chloride with stirring in an ice-
bath. To this 7.0 g. of 3-(methylamino)-
;
- ,~ . . . .
- . ., , : .:. : ~ :
: ~ . ::
. .
: : . ' :
: -
~z~
~ 2
-64-
propanoic acid, methyl ester is added ~ollo~,/ed
by N-methyl morpholine (5.5 ml.). After one hour
the ice-bath is removed and the reaction mixture
is kept at room temperature overnight. The reaction
mix-ture is then concentrated ln _acuo, taken up
into ethyl acetate and washed with 10~ potassium
bisulfate and saturated sodium bicarbonate
to yield 14.9 g. of crude product. Crystallization
from ether/hexane yields 10.7 g. of 1-[[(3-
methoxy-3-oxopropyl)methylamino]carbonyl]-L-
proline, l,l-dimethylethyl ester; m.p. 70-71.
c) l-[[(2-Carboxyethyl)methYlamino]carbonYl]-
L-proline, l,l-dimethylethyl ester
1~[[(3-Methoxy-3-oxopropyl)methylamino~-
carbonyl]-L-proline, l,l-dimethylethyl
ester (7.2 g.) is taken up into 47.7 ml. of
methanol to which 28.6 ml. of lN sodium
hydroxide is added with stirring. After 2.5 nours
the methanol is removed in vacuo. The aqueous
phase is acidified with dilute hydrochloric
acid and extracted into ethyl acetate to give
7.1 g. of crude product. Crystallization from
ether/hexane yields 6.1 g. of 1-[[(2-carboxyethyl)-
methylamino]carbonyl]-L-proline, l,l-dimethyl-
ethyl ester m.p. 69-71.
d) (+)-l-[[[4-(Benzoylamino)-3-oxo-5-PhenYlpentYl]-
methylamino]carbonyl]-L-proline~ dimethylethyl
ester
1-[[[(2-Carboxyethyl)methylamino]carbonyl]-
L-proline, l,l-dimethylethyl ester (900 mg.) is
,- :
. ,: : ~ .
i9~
-&,-
taken up into 10.5 ml. of ~etrahydrofuran wi~
stirring in an ice-bath. To this oxalyl
chloride (0.3 ml.) is added followed by
2 drops of dimethylformamide. After 20 minutes
the ice-bath is removed. After one hour at
room temperature the reaction mixture is
concentra~ed to dryness ln vacuo. The residue is
taken up into 6 ml. of tetrahydrofuran and while
stirring in an ice-bath 2-phenyl-4-(phenylmethyl)-
5(4H)-oxazolone (754 mg.) in 4.8 ml. of
tetrahydrofuran i5 added dropwise followed by
triethylamine (0.42 ml.). The reaction mixture
is kept at room temperature overnight, the
triethylamine hydrochloride salt is filtered off
and the filtrate is concentrated to dryness. T'ne
residue is taken up into 3.0 ml. of pyridine
and stirred for 3 hours with 9 mg. of 4-dime~hvl-
amino pyridine. Acetic acid (3 ml.) is added
and the mixture is heated at 100-105 for 30
minutes, concentrated ln vacuo, taken up into
ethyl acetate and washed with saturated sodium
bicarbonate and dilute hydrochloric acid
to yield l.l g. of crude product. Purification
on a silica gel column eluting with ethyl acetate:
hexane (2:1) gives 330 mg. of (~)-1-[[[4-(benzoyl-
amino)-3-oxo-5-phenylpentyl]methylamino]carbonyll-
L-proline, l,l-dimethylethyl ester.
:
- ~ 35
:`-
, ~
: :::. ::,
'- ' ~ ,......................... .
~: , :
~X~9
rl~s25
-66-
e) (+)-1-[[[4-(Benzo~Lamino)-3-_xo-5~en/l~,ent~
methylamino]carbonyl]-L-proline
The t-butyl ester product from pa-t (d)
(300 mg.) is treated for 1.5 hours with 3 ml. of
trifluoroacetic acid, concentrated in vacuo and
triturated to a solid with ether/hexane to
give 250 mg. o (+)-1-[[[4-(benzovlamino) 3-oYO
5-phenylpentyl]methylamino]carbonyl]-L~proline;
m.p. 38-68; [~]D3 = -9.16 (c = 1.2, methanol);
Rf = 0.71 [silica gel, chloroform:methanol:
acetic acid (9:0.5:0.5)].
Anal. calc d. for Cz5 29 3 5 2
C, 63.04; H, 6.72; N, 8.82
Found: C, 63~04; H, 6.29; N, 8.61.
15Example 66
(+)-1-[[[3-(Benzo~lamino)-2-oxohe~tyl]methylar.ino]-
.
car ~ L~~roline
a) N-[(Phenylmethoxy)carb~yl]sarcosine~ 1,1-
__ __
dime~hylethyl ester
~0 A solution of N-[(phenylmethoxy)carbonyl~-
sarcosine (114.5 ~.), methylene chloride ~250 ml.),
concentrated sulfuric acid (4 ml.) and isobutylene
1600 ml.) is shaken in a Parr shaker for 3 days
followed by neutral wash to aive 136.5 g. o~
~-[(~henylmethoxy)carbonyl]-sarcosine, 1,1-
dimethylethyl ester.
b) Sarcosine, l,l-dimethylethyl ester
N-[(Phenylmethoxy)carbonyl]-sarcosine, l,l-
diemthylethyl ester (68 g., 238 mmole) is taken
into absolute ethanol (500 ml.) and stirred under
`
- 35
`::
:, . - . . ,
~ ~ . . , -
.": '
-
`~
, , ~ , . . . . .
~9~ fl~4~1 ~
67-
hydrogen in the presence of 10?~ ~alladium on carbon
catalyst ~6.6 g.) overnight at room temperature.
The reaction mixture is then ~iltered to remove
the catalyst and concentrated in vacuo to remo-te
the ethanol and give 2~.6 g. of sarcosine,l,l-
dimethylethyl ester as an oil.
c) l-[[N-[[(l,l-Dimethylethoxy~carbon~l]methyl]
methylamino]carbonyl]-L-proline, phen~lmeth~l ester
__
L-Proline, phenylmethyl ester, hydrochloride
(2.41 g., 10 mmole) is taken into 40 ml. of
methylene chloride and N-methylmorpholine (2.3 ml.,
25 mmole) with stirring at -20. To this is
added dropwise 12.5% phosgene in benzene (16 ml.,
15 mmole). After 30 minutes at -20, the mixture
lS is concentrated to dryness in vacuo and taken into
40 ml~ of methylene chloride with stirring in an ice
bath. To this is added sarcosine, l,l-dimethyl-
ethyl ester (1.6 g., 11 rnmole) followed by
N-methylmorpholine (1.1 ml., 10 mmole). A~ter one
hour the bath is removed and the reaction mixture is
stirred overnight at room temperature, concentrated
in vacuo, taken inko ethyl acetate and washed neutral
with 10% po~assium bisulfate and saturated sodium
bicarbonate. The crude residue (3.6 g.) is purified
in 180 g. of silica gel in ethyl acetate:hexane
(1:1) to glve 2.7 g. of l-[[N-[[~l,l-dimethyl-
ethoxy)carbonyl]methyl]methylamino]carbonyl]-L-
proline, phenylmethyl ester.
................ ' .
. .
.
~ 1~.2
68-
d) 1-[[(2-Carboxyethyl)methylamino]carbonyl]-L-
roline, phenylmethyl ester
The ester product from part (c) (Z.7 g.,
7.17 mmole)iS treated or 1.5 hours with 10 ml.
of trifluoroacetic acid and 1.6 ml. o anlsole.
After concentrating to dryness it i5 triturated
with ether-hexane. The crude material is
crystallized from ether to yield 2 g. of
1-[[(2-carboxyethyl)methylamino]carbonyl]-L-
proline, phenylmethyl ester; m.p. 102-104.
e~ N-(Benzoyl)-D,L-norleucine
D,L-Norleucine (39.3 g., 300 mmole) is taken
into 150 ml. of 2N sodium hydroxide and while
stirring in an ice bath 150 ml. of 2N sodium
hydroxide and benzoyl chloride (38.3 ml., 330 mmole)
are added over a 30 minute period. The bath is
removed and after l.S hours the reaction mixture
is extracted with ether. The aqueous portion
is acidified with 2N hydrochloric acid and the
crystals filtered to give 68.9 g. of N-(benzoyl)-
D,L-norleucine m.p. (125) 131-133.
f) 2-Phenyl-4-butyl-5(4H)-oxazolone
N-(Benzoyl)-D,L-norleucine (40 g., 170 mmole)
is taken into 300 ml. of tetrahydrofuran with
stirring in an ice bath. To this is added
dropwise dicyclohexylcarbodiimide (38.52 g.,
187 mmole) in tetrahydrofuran ~195 ml.). After
15 minutes the bath is removed and the reaction is
allowed to run overniqht. The dicyclohexylurea
is filtered off and the filtrate is concentrated
3S
~:
:
:
~.~,69~
-69-
O to dryness. The crude product (31.7 ~.) is ~uri~ied
on silica gel in hexane:ether (2:1) to give
2-phenyl-4-butyl-5(4H)-oxazolone. This product
crystallizes neat when refrigerated.
g) (+)-1-[[[3-(Benzoylamino)-2-oxohe~ 1]-
methylamino]carbonyl] L-proline, ph ~lmethvl ester
1-[[(2-Carboxyethyl)rnethylamino]carbonyl]-L-
proline, phenylmethyl ester (4.8 g., 15 mmole) is
taken into 50 ml. of dry tetrahydrofuran with
stirring in an ice bath. To this is added droPwise
oxalyl chloride (1.58 ml., 1~ mmole) followed
by 4 drops of dimethylformamide. After 20 minutes
the bath is removed and the reaction is run for one
hour at room temperature before concentrating to
dryness. This material is taken into 30 ml. of
tetrahydrofuran, chilled and added dropwise to
2-phenyl-4-butyl-5(4H)-oxazolone (3.42 g., 15.75
mmole) in 24 ml. of tetrahydrofuran while stirring
in an ice bath. Triethylamine (2.55 ml.) is
added. After 5 minutes the bath is removed and
the reaction is run overnight at room temperature.
The triethylaminè hydrochloride salt is filtered
off, the filtrate is concentrated to dryness,
taken into 16 ml. of pyridine, 50 mg. of
4-dimethylamino pyridine is added, and the mixture
is stirred for 3 hours. Acetic acid (16 ml.)
is added and the mixture is heated at 100 for
45 minutes. The mixture is concentrated to
dryness, taken into ethyl acetate and washed
neutral with saturated sodium bicarbonate and
. . ,
.
.
:,. :, . ;:
- .,
. .
~, - - . , , :
~z~ 6 ~It~2~
-70~
dilute hydrochloric acid to yive 4.7 y. of
product. Purification on silica gel in benzene:
ethyl acetate (1:2) yields 1.14 g. of (~
[[[3-(benzoylamino)-2-oxoheptyl]methylamino]-
carbonyl]-L-proline, phenylmethyl es-ter.
h) (+)-1-[[[3-(8enzoylamino)-2-oxohept~l]-
methylamino]carbonyl]-L-proline
The ester product from part ~g) (650 my.)
is taken into 30 ml. of absolute ethanol containing
120 m~. of 10~ palladium on carbon catalyst
and reduced under hydrogen for 20 hours. The
reaction mixture is filtered and concentrated
to dryness to yield 500 mg. crude product.
Purification on a silica gel column with
lS chloroform:methanol:acetic acid (90:5:5) gives
340 mg. of (+~ [[[3-(benzoylamino)-2-oxoheptyl]-
methylamino]carbonyl]-L-proline; m.p. 40-80;
[~]D3~ -8.2~ (c z 1.1, methanol). Rf 0.52
(silica gel; chloroform:methanol:acetic acid;
90:5:5).
Anal. c 21 29 3 5 2
C, 60.19; H, 7.39; N, 10.03
Found: C, 60.19; H, 7.13; N, 10.3~.
:,, , ~ , . - ~ .
:.~ ', - ~,. . .
-71-
Exa ple 67
~ 1 [[[7-Amlno-3-~benzoylamino)-2-o~ohe~t~
methylamino]carbonyl]-L-proline
a) N -Benzoyl-N -~(phenylmethoxy)carbon~ L-
_ _ . _
lysine
N6-[(Phenylmethoxy)carbonyl]-L-l~sine
(20.1a g., 72 rnmole) is taken into 72 ml. of
lN sodium hydroxide with stirring in an ice bath.
To this over 20 minutes is added benzoyl chloride
(10.0 ml., 86.2 mmole) and 4N sodium hydroxide
(21.6 ml.). The bath is removed and the reaction
is allowed to run for 1.5 hours at room
tempera-ture. The mixture is extracted with ethyl
acetate, the aqueous portion is acidified wi-th
dilute hydrochloric acid, and extracted with ethyl
acetate. The ethyl acetate extract is concentrated
to low volume and hexane is added to crystallize
out 25.7 g. of N2-benzoyl-N6-[(phenylmetho~y)-
carbonyl]-L-lysine; m.p. 110-112.
.
b) 2-Phenyl-4-[4-[[(phenYlmethoxy)carbonYl]amino~
butyl~-5(4H)-oxazolone
q c
N~-Benzoyl-NU-[(phenylmethoxy)carbonyl]-L-
lysine (23.06 g., 60 mmole) is taken into 110 ml.
of dry tetrahydrofuran with stirring in an ice
bath. To this dicyclohexylcarbodiimide (13.6 g.,
66 mmole) is added dropwise in 70 ml. of dry
tetrahydrofuran. The bath is removed and the
reaction is kept at room temperature overnight.
The dicyclohexylurea is filtered off and the
filtrate is concentrated to dr~ness and crystallized
: .. . .. . ....
: : -: . .:
-: ~
- : .
-
,,
, ' ' ' ,
~2~9~ 259~ ~
-72-
from ethyl ace-tate:hexane to yive 21.4 y. of
2-phenyl-4-[4-[[lphenylmethoxy)carbonyl]amin
butyl]-5(4H)~oxazolone.
c) (+)-1-[[[3-(Benzo~lamino)-7-[[(ph2nylme~hox
carbonyl]amino]-2-oxoheptyl]meth~lamino]carbonyl]-
L- roline henvlmethvl ester
~ , P ~
1-[[(2-Carboxyethyl)methylamino]carbon~l]-L-
proline, phenylmethyl ester (6.4 g., 20 mmole) is
taken into 65 ml. of dry tetrahydrofuran with
stirring in an ice bath. Oxalyl chloride (2.1 ml.,
24 mmole) is added dropwise followed by 5 drops
of dimethylformamide. After 20 minutes the ice
bath is removed. After one hour at room temperature
the reaction mixture is concentrated to dryness
lS in vacuo. The residue is taken into 40 ml. Gf
dry tetrahydrofuran and added dropwise to N -benzo~l-
N6-[(phenylmethoxy)carbonyl]-L-lysine (7.7 g.,
24 mmole) in 30 ml. of dry tetrahydrofuran while
stirring in an ice bath~ Triethylamine (3.4 ml.,
24 mmole) is then added. The reaction is run over-
night at room temperature. The triethylamine
hydrochloride salt i5 filtered off, the filtrate
is concentrated to dryness, taken into pyridine
(21.2 ml.), 4-dimethylamino pyridine (66.6 mg.)
is added, and the mixture is stirred for 3 hours
at room temperature. Acetic acid ~21.2 ml.) is
added and the reaction is heated for 45 minutes
at 100. The reaction mixture is concentrated
to dryness, taken into ethyl acetate, and washed
neutral with saturated sodium bicarbonate and
- , ,
., . -~ . , .
, ' ' ,'
. -.~
2$9a
-73-
and dilute hydrochloric acid. The crude prod~ct~9.87 g.) is chroma-tographed twice on silica ae].
in ethyl acetate:ben~ene (2:1J to yield l.g y.
of (~-1-[[[3-(benzoylamino)-7-[[(phen~lmetho~
carbonyllamino]-2-oxoheptyl]methylaminolcarbon~
L-proline, phenylmethyl ester.
d) (~)-1-[[[7-Amino-3-(benzo~lamino)-2-o~ohept~l]
methylamino]carbonyl]-L-proline
The ester product from part (c) (950 mg.,
1.47 mmole) is taken into 100 ml. of 95~ ethanol
with stirring. Ammonium chloride (1.45 ml.) and
180 mg. of 10~ palladium on carbon catalyst are
added. The mixture is stirred overnight under
hydrogen, filtered, and concentrated to dryness.
The crude product is lyophilized from water to
give 613 mg. Purification on silica gel using
the solvent system chloroform:rnethanol:acetic acid
(8:2:2) gives 403 mg. of product which is then
applied to an AG50WX2 column and eluted with 2
aqueous pyridine to give 260 mg. of (~)-1-[[[7-
amino-3-(benzoylamino)-2-oxoheptyl~methylamirlo]
carbonyl]-L-proline, m.p. 130-133; [~] = -20.5
(c = 0.9, methanol). Rf 0.35 (silica geDl;
chloroform:methanol:acetic acid, 8:2:2).
Anal. calc'd. for C21H30 4 5 2
C, 57.78; H, 7.39; N, 12.84
Found: C, 57.70; H, 7.44; N, 12.84.
.
.
:
- :
i-lA25~a
-74-
(+)-1-[[[3-(Benzoylamino)-4~ -indol-3-y_)-2-
oxobutyl]methylamino]carbonyl]-L-proline
a) N-Benzoyl-L-tryptophan
L-Tryptophan (61.2 g., 300 mmole) is ta'~en into
600 ml. of 0.5 N sodium hydroxide with stirring
in an ice bath. Benzoyl chloride (38.3 ml., 330
mmole) and lN sodium hydroxide (330 ml.) are
added over a 25 minute period in 5 equal portions.
After 15 minutes the bath is removed and the reaction
proceeds for 2 hours at room temperature. The
reaction mixture is extracted with ethyl acetate
and the aqueous portion is acidified with concentrated
hydrochloric acid and extracted into ethyl ace'ate.
The crude product (105 g.) is crystallized from
ether to yield 103.3 g. of N-benzoyl-L-tryptophan;
m.p. 84-86 (an ether adduct).
b) 2-Phenyl-4 [(lH-indol-3-yl)meth~1]-5(4H)-
oxazolone
N-Benzoyl-L tryptophan (50 g., 130.74 mmole)
is taken into 200 ml. of tetrahydrofuran with
stirring in an ice bath. Dicyclohexylcarbodiimide
(27 g., 130.74 mmole) in 60 ml. of tetrahydrofuran
is added dropwise. After 15 minutes the ice bath is
removed and the reaction proceeds overnight. The
dicyclohexylurea is filtered off and the filtrate is
concentrated to dryness in vacuo. The crude product
is crystallized from methanol to give 30.18 g. of
2-phenyl-4-~(lH-indol-3-yl)methyl]-5(4H)-oxazolone;
m.p. 141-143.
:
,
,- . ~ :
- ~ - ,
~' ' ' ' . ' . - . ' : ,
' -
~. : . .
J~l .
-75-
c) (+)-1-[[[3-(senzo~l~mino)-4-~ -indol-3-
yl)-2-o~obutyl]methylaminolcarbon~ _-proline,
phenylmethyl ester
1-[[~2-Carboxyethyl)methylamino]carbonyl~-L-
proline,phenylmethyl ester (3.2 g., 10 r~ole) is
taken into 33 ml. of dry tetrahydrofuran wit'n
stirring at 5. Oxalyl chloride (1.05 ml., 12 r~nole)
is added dropwise followed by 4 drops of dimethyl-
formamide. After 20 minutes the ice bath is
removed and after stirring for one hour at
room temperature the reaction mixture is
concentrated to dryness , taken into 20 ml. o~
tetrahydrofuran, chilled and added dropwise to
2-phenyl-4-[(lH-indol-3-yl)methyl]-5(4H)-oxazolone
(3.0 g., 10.5 mmole) in 16 ml. of dry tetrahydro-
furan with stirring in an ice bath. Triethyl-
amine (1.7 ml., 12 mmole) is then added to the
reaction mixture. After 15 minutes the ice bath
is removed and the reaction is run overnight at
room temperature. The triethylamine hydrochloride
salt is filtered off and the iltrate is concentra-
ted to dryness, taken into 10.6 ml. of pyridine,
33.3 mg. of 4-dimethylamino pyridine is added, and
the mixture is stirred for 3 hours. Acetic acid
(10.6 ml.) is added and the reaction is heated
for 45 minutes at 100 under an argon atmosphere.
The reaction mixture is concentrated to dryness,
taken into ethyl acetate, and washed with
saturated sodium bicarbonate and dilute hydro-
30 ~ chloric acid. The crude product (4.6 g.) is
. ~ ..
. ............... .
. ~ . ..
: :
9~
.2.-~g,~
16-
purified on a silica gel column in ethyl acetate:benzene (2:1) to yield 523 mg. of (+)-l-[[[~-(benz-
oylamino)-4-llH-indol-3-yl)-2-oxobut~l~methyl~mino~-
carbonyll-L-proline phenYlmethyl ester.
d) (+)-1-[[[3-(Benzoylamino)-4-(lH-inclol-3~ 2-
_xobutyl]methylamino]carbonyl]-L-proline
The ester product from part (c) (520 mg.
1 mmole) is taken into 25 ml. of 95% ethanol
containing 100 mg. of palladium on carbon catalyst
(10%) and the mixture is stirred under positive
hydrogen pressure overnight. The reaction mixture
is filtered and concentrated to dryness. The
crude product (400 mg.) is purified on a silica
gel column with chloroform:methanol:acetic acid
(90:5:5) to give 212 mg. of ( )-1-[[[3-(benzoyl-
amino)-4-(lH indol-3-yl)-2-oxobutyl]methylamino]-
carbonyl-L-proline; m.p. 102-128; [~]D -1.3
(c =-0.91 methanol). Rf 0.41 (silica gel; chloro-
form:methanol:acetic acid 90:5:5)
20 Anal. calc d- for C26 28 4 5 2
C 63.23; H 6.11; N 11.35
Found: C 63.23; H 5.92; N 11.01.
Example 69
(~)-1-[ [ [3-(BenzoYlamino)-4-(4-hydroxyphenyl)-2
oxobutyl]methylamino]carbonyl]-L-proline
a) 2-Phenyl-4-[[4-(phenylmethoxy)phenyl]methll]
5(4H)-oxazolone
O-Benzyl-L-tyrosine (11.0 g. 40.5 mmole)
is taken into 0.5 N sodium hydroxide (81 ml.)
and water (81 ml.) with vigorous stirring in an
'~`
. ~ 35
`...... :
,, ,
: . . ` :, . ` ,
:: . - .:~,, , ~ . .
~.2
-77-
ice-bath. To this in five equal portions i3 added
a total of 52 ml. of benzoyl chloride, 45 ml. of
lN sodium hydroxide and an additional 400 rnl. of
water over a 25 minute period. The bath is
removed and the reaction is run for 2 hours at
room temperature. The mixture is extracted t./ice
with ethyl acetate. The aclueous portion is filtered,
acidified with lN hydrochloric acid and the crystals
filtered to give 12.9 g. of N-benzoyl-O-benzyl-L-
tyrosine; m.p. 166-168 (162).
This N-benzoyl-O-benzyl-L-tyrosine (12.76 g.,
35 mmole) is taken into dry tetrahydrofuran (50 ml.)
with stirring in an ice-bath. To this dicyclohexyl-
carbodiimide (7.7 g., 37.4 mmole) in tetrahydrofuran
(18 ml.) is added dropwise. After 20 minutes,
the ice-bath is removed and the reaction proceeds
overni~ht at room temperature. The dicyclohexyl-
urea is filtered off and the filtrate is concentrated
to dryness. The crude product is crystallized
from ether/hexane to give 10.26 g. of 2-phenyl-
4-[[4-~phenylmethoxy)phenyl]methyl~-;(4H)-
oxazolone; m.p. 85-87 (83).
b) (+)-1-[[[3-(BenzoYlamino)-2-oxo-4-[4-!~henYl-
methoxy)phenyl]butYl~methylamino]carbonyl~-L-
Droline, phenylmeth,yl ester
1- [[(2-Carboxyethyl)methylaminc]carbonyl]-L-
proline, phenylmethyl ester (6.4 g., 20 mmole)
is taken into 65 ml. of dry tetrahydrofuran ~ith
stirring in an ice bath. Oxalyl chloride (2.1 ml.,
24 mmole) is added dropwise followed by 15 drops of
`:
`: :
, .
:i
.
...... . .
.
;:: . :
- ~ ;' ' ,' :
:
~IA25s,~
-78-
O dimethylformamide. After 20 minutes the ice
bath is removed and the reaction proceeds for
one hour at room temperature. The reaction
mixture is concentrated to dr~ness, taken into
40 ml. of tetrahydrofuran and added drop~,/ise to
a solution of 2-phenyl-4-[[4-~phenylmethoxy)
phenyl]methyl]-5(4H)-oxazolone (7.5 g., 21 mmole)
in 30 ml. of tetrahydrofuran while stirring in an
ice bath. Triethylamine (2.8 ml., 20 mmole)
is then added to the reaction mixture. After
30 minutes the ice bath is removed and the reaction
is run overnight at room temperature. The
triethylamine hydrochloride salt is filtered off and
the filtrate is concentrated to dryness ln vacuo.
The crude residue is taken in 21 ml. of pyridine,
67 mg. of 4-dimethylamino pyridine is added, and
the mixture is stirred for 3 hours under an argon
blanket. Acetic acid (21 ml.) is added and the
reaction mixture is heated ~or 45 minutes at 100
with stirring under a constant stream of argon.
The reaction mixture is concentrated to dryness,
taken into ethyl acetate and washed neutral with
saturated sodium bicarbonate and dilute hydro-
chloric acid. The crude ~roduct (11.9 g.) is
chromatographed over 600 g. of silica gel with
ethyl acetate:benzene (2:1) to yield 4.6 g. of
(+)-l-[[[3-(benzoylamino)-2-oxo-4-[4-(phenylmethoxy)
phenyl]butyl]methylamino]carbonyl]-L-proline,
phenylmethyl ester.
.
: ::
: .- . . .
- , - ~:
- ,
~2~
~ .,_
HA259a
~79-
c) (~-1-[[[3-(Benzoylamino?-4-(4-hydrox~phenYl)-
2 oxobutyl]methylamino~carbonyll-L-pro_in
The ester product from part (b) (2 g.,
2.83 mmole~ is ~aken into 125 ml. o~ methanol
containing 400 mg. of palladium on carbon catalyst
(10%) and stirred under positive hydrogen ~ressure
for 20 hours. The reaction mixture is filtered
and concentrated to dryness in vacuo. The crude
product is purified on a silica gel column with
chloroform:methanol:acetic acid (90:5:5) to yield
1 g. of product. This is ~aken into chloroform
and extxacted with water. The chloroform solution
is evaporated to give 415 mg., of t~)-1-[[[3-
(benzoylamino)-4-(4-hydroxyphenyl)-2-oxobutyl]-
methylamino]carbonyl]-L-proline; m.p. (103) 120-146,
[a]D ~35 (c = 1.0, methanol). R~ 0.48 (silica
gel; chloroform:methanol:acetic acid, 9:1:1).
Anal. calc d- for C24 27 3 6 0.4 H2 :
C, 62.57; H, 6.08; N, 9.12
Found: C, 62.64; H, 6.11; N, 9.04.
Example 70
(+)-l-[t[3-(BenzoYlamino)-2-oxo-4-(3_pyridin~1)bu~yl]
methYlamino]carbonyl]-L-Proline
a) 2-(BenzGylamino)-3-(3-pyridinyl)-2-propenoic
acid
3-Phenyl-4-(3-~yridinylmethylene)-5(4H)-
oxazolone (3 g., 12 mmole) tsee Grif~ith et al.,
J. Org. Chem., Vol. 29, p. 2659] is dissolved in
acetia acid (24 ml.) and aqueous hydrochloric
acid (0.5 N, 150 ml.). The reaction ~ixture i~
.
~ ' :
:
~ 35 ~
: `
~`~
, ~ :
, ~ :
~, :
HA259a
_-- 80--
stirred overnight at room temperature. It is
evaporated and reevaporated ~rom absolute ethanol.
It is triturated with tetrahyclrofuran, filtered,
and the filtered solid is retriturated with
S absolute ethanol to yield 2.8 g. of 2-(benzoylamino)-
3-t3-pyridinyl)-2-propenoic acid; m.p.
215-216 (203).
b) 2-_Benzoylamino)-3-(3-pyridinyl)-pro~anoic acid
2-(Benzoylamino)-3-(3-pyridinyl)-2-3ropenoic
acid (14 g., 46 mmole)is dissolve~ in water (500 ml.)
and hydrogenated using palladiurn on carbon catalyst
(10%, 1.8 g.) overnight. The catalyst is filtered
off, and the reaction mixture is evaporated to a
small volume (100 ml.) and lyophilized to give
13.1 g. of product. ThQ lyophilate is triturated
with absolute ethanol-ether mixture and filtered
to give 12 g. of 2-(benzoylamino)-3-(3-~yridinyl)-
propanoic acid; m.p. 99-115.
c) (~)-1-[~[3-(Benzoylamino)-2-oxo-4-(3-pyridinyl)-
bUtYl]methylaminolcarbonyl]-L-proline, phenylmethyl
ester
2-(Benzoylamino)-3-(3 pyridinyl)-propanoic acid
(3 g., 9.8 mmole) is suspended in tetrahydrofuran
(30 ml.) and while stirring in an ice bath
triethylamine (1.4 ml., 10 mmole) and dicyclo-
hexycarbodiimide (2.1 g., 10.2 mmole) are added.
The reaction mixture is stirred at room temperature
overnight. It is then filtered and the iltrate
is evaporated to dryness. ~his oxazolone is then
dissolved in tetrahydrofuran (15 ml.).
; .
: ., , :: - .
.
- ~ -. . -. .::
~: .
.,. : ',
~6~
. HA259a
~1-
1-[[(2-Carboxye~hyl)methylamino]carbonyl~-L-
proline, phenylmethyl ester (3 g., 9.4 mrnole) is
taken into dry tetrahydrofuran and treated with
oxalyl chloride and dimethylformamide as set
forth in Exarnple 66 (g). The resulting acid
chloride is taken into tetrahydrofuran (15 ml.),
chilled, and added dropwise to the above oxazolone
tetrahydrofuran solution while stirring in an ice
bath. Triethylamine (1.6 ml., 11.4 mmole) is
added and the reaction mixture is s~irred at
room temperature overnight. Triethylamine hydro-
chloride salt is filtered off and the filtrate is
evaporated in vacuo. The concentrated residue
is redissolved in pyridine ~10 ml.), 4-dimethylamino
pyridine ~50 mg.) is added, and the solution is
stirred at room temperature ~or 3 hours. Acetic
acid ~11 ml.) is then added and the reaction
mixture is heated at 100 for 40 minutes. It is
then evaporated, redissolved in ethyl acetate and
extracted with aqueous saturated sodium bicarbonate
solution followed by water. The remaining ethyl
acetate extract is concentrated and chromatographed
over siIica gel using the solvent system ethyl
acetate:methanol ~95:5) to give 0.8 g. of (+)-1-
[~[3-(benzoylamino)-2-oxo 4-(3-pyridinyl)butyl]
methylamino]carbonyl]-L-proline, phenylmethyl ester.
'
., :: .
~ . ;,, ~, ;
-' .: .
. . .
. :., .
~l~6~
HA259a
~2-
d) (~ [[[3-(Ben~oylamino)-2-oxo-~ eyridinyl)_
butyl]methylamino]carbonyl]-L-Proline
The es~er product from part (c) (0.7 y., 1.32
mmole) is dissolved in ethanol (50 ml.).
Palladium on carbon catalvst llO~, 100 mg.)
is added and the solution is stirred under
an atmosphere of hydrogen overnight. It
is filtered and the filtrate evaporated
tO.6 g.). This material is chromatographed
over silica gel using the solvent system
chloroform:methanol:acetic acid ~8:1:1)
to yield 0.35 g. of crude product. This
is combined with additional material (0.15 g.)
of similar purity from another ruin of
the reaction andthe combined material
tO.5 g.) is applied to a AG-SO(H )
column (10 ml. bed volume). The column
is washed with water (lO0 ml.) and then the
product is eluted out with 2~ aqueous
pyridine. The fractions containing the
product are pooled, evaporated,
.: - . , ~ . ,- :.
. . . . . .
- .
' ' ` .
HA259a
-83-
redissolved in water and lyophilized to give
0.38 g. of (~ [[3-(benzoylamino)-2-
oxo-4- ( 3-pyridinyl)butyl]methylamino]
carbonyl]-L-proline: m.p. 92-120;
[a]D = ~ 6.9 (c = 1.1, methanol).
Rf 0.24 (silica gel; chloroform:methanol:
acetic acid 8:1:1)
23 26 4 5 0.9 H2O:
C, 60.75; H, 6.16; N, 12.32
Found: C, 60.74; H, 5.91; N, 12.35.
Example 71
1-~[(4-Aminobutyl)[3-5benzoylamino)-2-
oxo-4-phenylbutyl_amlno]carbonyl]-L
Proline, monohydrochloride
a) N-[(Phen~lmethoxy)carbon~1]-4-
h~ydroxybutylamine
4-Hydroxybutylamine (10 g.,
112.18 mmole) is taken into 200 ml.
of dry tetrahydrofuran with stirring
in an ice bath. Triethylamine (17.2 ml.,
123.4 mmole) is added dropwise followed
, . . . .
,. : ~ i :
~ .. i .
:: , . - . -
':~. - .
HA259a
-84-
O by phenylmethoxycarbonyl chloride (17.6 ml., 123.4
mmole). After one hour the bath is removed and
the reaction proceeds overnight at roorn temperature.
The mixture is concentrated to dryness in vacuo,
taken into ethyl acetate and washed with water.
The crude product is crystallized from ethyl
acetate/hexane to give 16.34 g. of N-[(phenyl-
methoxy)carbonyl]-4-hydroxybutylamine.
b) N-[(Phenylmethoxy)carbonyl]-4-bromobutylamine
N-~(phenylmethoxy)carbonyl]-4~hydroxybutyl-
amine (16.07 g., 72 mmole) and triphenylphosphine
(20.75 g., 79 mmole) are taken into 105 ml. of dry
tetrahydrofuran into a 3-necked flask stirring
and fitted with a condenser. N-Bromosuccinimide
(14.1 g., 79 mmole) is added portionwise over
a 10 minute period. After 45 minutes the mixture
is concentrated to dryness, crystallized from hexane:
ethyl acetate (3:1) to remove tri~henylphosphine
oxide, and the mother liquor is concentrated to
dryness to yield 20 g. of crude product. This is
purified on 200 g. silica gel in hexane:ethyl
acetate (3:1) to yield 14.1 g~ of N-[~phenylmethoxy)-
carbonyl~-4-bromobutylamine.
.
c) N-[[[(PhenvlmethoxY)carbo~yl]aminolbutYll-
glycine,l,l-d m thylethyl ester
N-[(Phenylmethoxy)carbonyl]-4-bromobutylamine
(6.86 g., 24 mmole) and glycine, l,l-dimethylethyl
ester (4.722 g., 36 mmole) are taken into 48 ml.
of dimethylformamide. ~riethylamine (3.36 ml.,
24 mmole) is added to the reaction mix-ture.
:
,~ .
. . ~ -
- .
.
,
. .
HA259a
-85_
After 72 hours at room temperature, sodium
bicarbonate (6 g.) is added and after 1.5 hours
the mixture is filtered and concentrated to
dryness ln vacuo. The residue is purified on silica
gel in methanol:ethyl acetate (5:95) to give
5.63 g. of N-[[[(phenylmethoxy)car~onyl]amino]-
butyl]glycine, l,l-dimethylethyl ester.
d) l~[[[(l,l-Dimethylethoxv)carbonyllmethyl]
[[[(phenylme hoxy)carbon~l]amino]butyl]aminol
carbonyl]-L-proline, phenylmethyl ester
L-Proline, phenylmethyl ester, hydrochloride
(2.364 g., 9.81 mmole) is taken into 39 ml. of
methylene chloride with stirring at -20. N-
Methylmorpholine (2.76 ml., 24.48 mmole) is added
followed by the dropwise addition of phosgene
(14.7 mmole, 15.6 ml. of 12.5~ solution in benzene).
After 30 minutes at -20, the mixture is concentrated
to dryness ln vacuo. The residue is taken into
30 ml. of methylene chloride and added dropwise
to N-[[[(phenylmethoxy)carbonyl]amino]butyll-
glycine,l,l-dimethylethyl ester (3.0 g., 8.916
mmole) in 6 ml. of methylene chloride with
stirring in an ice bath. N-Methylmorpholine
(1.08 ml., 9.8 mmole) is added and after one hour
the bath is removed and the reaction proceeds
overnight at room temperature. The mixture is
~concentrated to dryness Ln vacuo, taken into
ethyl acetate and washed neutral with 103 potassium
bisulfate and saturated sodium bicarbonate. The
-
- 30 crude product i~ puri~ied on 125 g. of silica gel
.
.
. .
-': : ,
.
HA259
-~6-
in ethyl acetate:cyclohexane (2:1) to give 5.7 g.
of l-[[[[(~ dimethylethoxy)carbonyl~meth
[[[(2henylmethoxy)carbonyl]amino]butyl~amino~
carbonyl]-L-proline, phenylmethyl ester.
e) 1-[[(Carbox~methy~)[[[(pheny_methoxY)carbon~l]
amino]butyl]amino]carbonyl]-L-proline, phenylme~hYl
ester
The ester product from part (d) (5.0 g.,
8.8 mmole) is treated for 1.5 hours with 25 ml. of
trifluoroacetic acid and 2.1 ml. of anisole,
concentrated to dryness and treated twice with
cold ether/hexane and decanted. The crude
product (4.7 g.) is purified on 250 g. of silica
gel in chloroform:methanol:acetic acid (90:5:S)
lS to give 2.77 g. o l-[[(carboxymethyl)[[[(phenyl-
methoxy)carbonyl]amino]butyl]amino]carbonyl]-L-
prolinetphenylmethyl ester.
f) 1-[~[3-(BenzoYlamino)-2-oxo-4-E~enYlbutvl]
[[[(Phenylmethoxy)carbonyl]amino]butyl]amino]
carbonyl]-L-proline, phenylmethyl ester
The ester product form part (e) (2.7 g.,
S.27 mmole) is taken into 17 ml. of dry tetra-
hydrofuran with stirring in an ice bath. Oxalyl
chloride (0.55 ml., 5.8 mmole) is added dropwise
followed by the addition of 4 drops of dimethyl-
formamide. After 20 minutes the bath is removed
and after an additional hour the mixture is
concentrated to dryness. The residue is taken
into 10 ml. of te~rahydrofuran with stirriny in an
ice bath. 2-Phenyl-4-phenylmethyl-5(4H)-oxa~olone
:
. .
.` : .. .
:
-
~2~ 6
HA259a
-87-
(1.33 y., 5.27 mmole) in 8 ml. of tetrahydro~uran
is added dropwise followed by triethylamine
(0.9 ml., 6.42 ml.). Af~er 30 minutes the bath
is removed and the reaction proceeds at
room temperature. The trie~hylamine hydrochloride
salt is filtered off and the filtrate is
concentrated to dryness ln vacuo. The residue
is stirred under an argon blanket in 5.6 ml.
of pyridine and 25 mg. of 4-dimethylamino pyridine
for 3 hours at room temperature. Acetic acid
(5.6 ml.) is added and the mixture is heated at 100
for 45 minutes under a gentle argon flow. The
mixture is concentrated to dryness, taken into
ethyl acetate a~d washed neutral with saturated
lS sodium bicarbonate and dilute hydrochloric acid.
The crude product is purified on a silica gel
column in ethyl acetate:benzene (1:1) to give
450 mg. of 1-[[[3-(benzoylamino)-2-oxo-4-
phenylbutyl][[[(phenylmethoxy)carbonyl]amino]
butyl]amino]carbonyl]-L-proline, phenylmethyl
ester.
g~ l-[[(4-Aminobutyl)[3-(benzoYlamino)-2-oxo-
4-phenylbutYl]amlno]carbonyl]-L-proline, mono-
hydrochloride
The ester product from part (f) (420 m~.,
0.584 mmole) is taken into 40 ml. of 95% ethanol,
0.584 ml. of lN hydrochloric acid, and 50 mg. o~
palladium on carbon catalyst (lO~ he mixture
is stirred under hydrogen pressure for 24 hours.
The reaction mixture i5 then fiitered and
.
,,: '. . :., ' ~
.
.: . .
-
. .
HA259a
-88-
concen~rated ~o dryness ln vacuo. The crude
product is puri~ied on an LH-20 column in water
to yield l9S mg. of L-[[(4-aminobutyl)[3-
(benzoylamino)-2~oxo-4-phenylbutyl~amino~carbonyl~-
L-proline, monohydrochloride; m.p. (92) 122-162;
[]23 +9~4O (c = 1.06, methanol).
Rf 0.34 (silica gel; chloroform:methanol:acetic acid;
8:2:2).
Anal- calc'd- ~or C27H34N405 ~ HCl 1-0 H2O
C, 58.99; H, 6.78; N, 10.19; Cl, 6.45
Found: C, 58.99; H, 6.56; N, 10.19; Cl, 6.59.
ExamPles 72 - 96
Following the procedure of Examples 65-~
the amino or imino acid ester shown in Col. I
is reacted with the oxazolone of Col. II to
yield the ester product of Col. III. Removal
of the R6 ester group yields the products wherein
R6 is hydrogen.
:
- ~ . .
-: ,
- : : ,
HA2 5 9a
--89--
Col. I
R O
~1 11
(C 2)n N C X
s
Col. II
N
~ \
R2-C C-R3
O ---C=O
Col. III
O R O
11 11 11
R3 CH-C- ( CH2 I n~N ---C-X
NH
C=O
R2
,
.
;
35:
. : :
, . ., ~ :
. . :
,
HA~5ga
--90--
.
S r~
U~ U~ (~5 (~ 5
N
,~` 15 y
~0 ~ ) U
2 5 ~ ~r
~ I
U~
~30
.~
X
:
3 5
` ~ :
'' ' ~ ~ , . ~ ' ' ' :, '
~ ~ , . . . . .
HA259a
-91--
S
u
u o o
10 Y 1 ~
Z--U
I
1 ~ u
25 ~
U r~
~ . U~
"
Ur~
:3 0: ' ~ ~
: ~ ~ ,, '
3 5 ~ ~
.
: - . -., : . :
.:
.
~IL26~
. ~lA2 53,~
--92--
~r7 :~
= ~ = X.~
~ _
~ ~/ \
O~
~,~ I U~ Z Z
Z
0~ .
2 5~ N
~ : 1 _
C)
: ~ 30 : ;~
t~ 03 CO
~` ~ W
:` ~ :
,
:`- ~: 35~
.
- ~
.. ~ :. . . .. .
- .
~2~9~6 ~--~
HA2 S9a
--~3--
~ ~ ", r~
lo x ~ 8 h
1 5 ~ v
v~
2 0 s~ v
v~
30:
G~
_~
I 6~ co a~
: ; 35
':
:
, :,. .
:
. :
.:
::. - ~ . . ...
: ~
:
,
~: .
-
~l~6~ 6 ~
- H~259~ -
--94--
1~ x I ; ~ s' Z~ =o
1 5
:
:
:
: :
:: :25 ~
: :
~ a
:;f ~ 0
: 3 0
Q. :
~9 1`CO :
:
: 3 5 ~
~z~
HA2 59a
95--
o ~ 8 ~
U (~ U-u-u
x I Z~ z- æ æ
~ l ~
u u ~
,........ ...
u
.
~ co o
~ ~ a~
:
` : 35
~- :
`, . ': :'
.
. . - : , . . .
. - : ' :. . . : , ,:,
~Z~ 6 ~
HA255~ -
--96--
3 U~
U r
0_ U O ~
~ ~: U~ U--U
UU -- o _ j ~1 G = I ~
xl z ~u~ q
~ U u
:`
~ N1 (~) $ (~)
~~
~_~ U
~ ~ X~
;
a~
~ x
:' '~ . ~ : `:
~z~
~ IlA25g~
--97--
~,
U
C~ 3
o--~ U
,o o = c)
U ~ _ U
,o o
o=~ ',~ o=y
10 Xl C. ¢
~ I U~ U",
.
- 2Q
.
: l~
U~
Z [~u~
: 3 0
: ~
` : 3 5 ~ ~
:. : ,- . ,.. :
:, ~ ~ - ,- , .: - : .
- : . - : . . .
.
~, ,, . ~ ... . .
, . ., ` : .
,
.. . HA25~a
--98--
The Rl protecting yroups in Examples 17,
79 and 80 ~ the R3 protectiny groups in
Examples 81 and 83 ~ and the R5 protecting groups
in Examples 84 and ~6 to 88 are removed as
the last step in the synthesis. The R6 ester
groups shown in Examples 93 to 96 are not
removed.
_ample 97
t)-1-[[[3-(Benzoylamino)-2-oxo-4-phenYlbutYl~-
methylamino]carbonyl]-L-proline, ethyl ester
~1)-1-[[[3-(Benzoylamino)-2-oxo-4-
phenylbutyl]methylamino]carbonyl]-L-proline
(1 g.) is treated for 7 hours at room temperature
with 10 ml. of 2N ethanol:hydrochloric acid,
concentrated ln vacuo, taken up into ethyl
acetate and washed neutral with 10~ potassium
bisulfate and saturated sodium bicarbonate to
yield 900 mg. of crude product. This materialis
purified on silica gel column eluting with
chloroform:methanol:acetic acid (90:3:3) to give
671 mg. of (+)-1-[[[3-(benzoylamino)-2-oxo-4-
phenylbutyl~methylamino]carbonyl]-L-proline,
ethyl ester; m.p. 40 - 60; Rf [chloroform:
methanol:acetic acid (90:3:3:]= 0.62.
Anal. calc'd. for C26H31N3O5
C, 66.24; H, 6.77; N, 8.91
Found: C, 66.24; H, 6.75; N, 8.79.
J
:` :
:;
;~ 35 ~
J~
HA25ga
99
In a similar manner, ethyl or other alkyl
esters of the compounds of Examples 2 to 58 and
65 to 92 can be prepared.
Example 98
t+)-l ~[[3-(Benzoylamino)-2-oxo-4-phenylbutyll
methylamino]carbonyl]-L-proline, 3-pyridinylmeth~l
ester, monohydrochloride
(~)-1-[[[3-(Benzoylamino)-2-oxo-4-phenylbutyl]
methylamino]carbonyl]-L-proline (1.312 g., ~ mmole),
4-dimethylamino pyridine (183 mg., 1.5 mmole),
3-pyridinylcarbinol (0.29 ml., 3 mmole), and
dicyclohexylcarbodiimide (618 mg., 3 mmole) are
taken into tetrahydrofuran (10 ml.) with stirring
in an ice bath. The reaction proceeds overnight
at room temperature. The dicyclohexylurea is
filtered off and the filtrate is concentrated to
dryness. The crude product is chromatographed on
silica gel in chloroform:methanol:acetic acid
(90:3:3) to yield 630 m~. of (+)-1-[[[3-(benzoyl-
amino)-2-oxo-4-phenylbutyl]methylamino]carbonyl]-L-
proline, 3-pyridinylmethyl ester.
This ester product (0.46 g., 0.87 mmole) is
dissolved in methanol (5 ml.) and aqueous lN
hydrochloric acid (0.87 ml.) is added. This
mixture is evaporated, dissolved in water,
filtered, and lyophilized to give 450 mg. of
(+)-l-[[[3-(benzoylamino)-2-oxo-4-ohenylbutyll
methylamino]carbonyl]-L-proline, 3-pyridinylmethyl
ester, monohydrochloride; m.p. (62) 88-155.
Rf 0.33 (silica gel; chloroform:methanol:
acetic acid; 90:3:3).
. ~
, . . ~ " , ~ . : , . . .
:
~z~
Hh259a
-1()0--
Anal. Calc'd. for C30H32N~Os HCl 1.45 H~O:
C, 60.94; H, 6.12; N, g.48; Cl, 6.00
Found: C, 60.94; H, 5.86; N, g.52; Cl, 5.92.
Examples 99-110
Following the procedure of Example 98, (~)-1-
[[[3-(ben~oylamino)-2-oxo-4-phenylbutyl]methylamino]-
carbonyl]-L-proline is treated with the reagent
shown below in Col. I to yield the ester product
shown in Col. II.
Col. II
O O o ~
~ gH C-CH2-N-C-N ~ CLO)OR6
15 Example ~ Col. I R6
O O
Il 11
99 Cl-CH-O-~C-C2H5 -,H_o-c-c2H5
bo ~o
Il 11
100 Cl-CH-O-C-C2H5 -CH-O-C-C2H5
CH(CH3) 2 CH(CH3) 2
O O
Il ll
101 Cl-CH2-O-C-C(CH3)3 -CH2-O-C-C(CH3)3
O O
102 Br-CH2-O-C-CH3 -CH2-O-C-CH3
:
, ~ . ..
,
' ~' ' ,
HA259a
-101--
Example Col. I R,
O O
10 3 Cl-CE{2-O-C ~> -CH -O-C
O O
104 I-CH2-C-O-C (CH3) 3 -CH -C-O-C (CH )
CE~ O CH O
1 3 11 1 3 11
105 I-C C-O-CH -C--C-O-CH3
l l
`~ CH3 CH3
.
5 106 CH-(CH2-0-CH2~ )2 -CH(CH,~-OH)2
OH
107 c~2- CH -- 2 -CH2-CH CH2
OH O O OH O~l
H2C~) H2C ~)
.
.
: `
~`
~ ~ 35 ~
. .
:
. ~ ~ : . : .
,, , ~
-. ~
6 ~ ~
HA259a
-102-
Example Col. I R6
10 8 HO- (CH2) 2-M (CH3) 2 - (CH2) 2M (CH3) 2
10 9 HO-CH2~ 2~1
110 Ho-CH2i~l -CH2~
In a similar manner, esters can be prepared
of the products of Examples 2 to 58 and 65 to 92.
` 30
.
::`
:;
.
:, . ' ' -:
,
: . - ' ` - : ,
. '. . .: - , .
~2~
HA259a
-103-
(+)-1-[[[3-(Benzoylamino)-2-oxo-4-phen~lbut~
methylamino]carbonyl]-L-proline, sodium salt
(+)-1-[[13-(Benzoylamino)-2-oxo-4-phenyl-
butyl]methylamino]carbonyl]-L-proline (1 mmole)
is dissolved in water (50 ml.). Aqueous sodium
bicarbonate (0.1 N, 20 ml.) is added and the
aqueous solution is lyophilized. It is then
dissolved in water (10 ml.) and applied on a
column ~5cm. x 60 cm.) of Sephadex chromatography
gel G-10 and eluted with water. Fractions
containing the desired product are pooled and
lyophilized to obtain (+)-l-[[[3-(benzoyl-
amino)-2-oxo-4-phenylbutyl]methylamino]carbonyl]-
L-proline, sodium salt.
ExamPle 112
1000 tablets each containing the following
ingredients
~ (+)-1-[[[3-(Benzoylamino)-2-
oxo-4 phenylbutyl]methylamino]-
carbonyl]-L-proline, sodium
salt 100 mg.
Corn starch 50 mg.
Gelatin 7.5 mg.
Aviceltmicrocrystalline
cellulose) 25 mg.
Magnesium stearate 2.5 mg.
are prepared from su~icient bulk quantities by
mixing the (~ [[[3-(benzoylamino)-2-oxo 4-
phenylbutyl]methylamino]carbonyl]-L-proline,
~:
--
, :` :
~ 35
:
,~. . ; "
. ~ -: : :: . . , :
: - .:
-: -
:lZ~ 9~ D
HA259a
-104-
sodium salt and corn starch with an aqueous
solution of the gelatin. The mixture is dried
and ground to a fine powder. The Avicel and
then the magnesium stearate are admixed with
granulation. This mixture is then compressed
in a tablet press to form 1000 tablets each
containing 100 mg. of active ingredient.
In a similar manner, tablets containing
100 mg. of the product of any of Examples 1
to 110 can be prepared.
A similar procedure can be employed to
form tablets containing 50 mg. of active
ingredient.
Example 113
lS Two piece # 1 gelatin capsules each containing
50 mg. of (~)-1-[[[4-(benzoylamino)-3-oxo-5-
phenylpentyl]methylamino]carbonyl]-L-proline,
sodium salt are filled with a mixture of the
following ingredients:
; 20 (~ [[[4-(Benzoylamino)-3-
oxo-5-phenylpentyl]methylamino]-
carbonyl]-L-proline, sodium
salt - 50 mg.
Magnesium stearate 7 mg.
~ 25 Lactose 193 mg.
`` 250 mg.
In a similar manner cap~ule~ cont~lning
50 mg. of the product of any of Examples 1 to
110 can be prepared.
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HA259~ -
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ExamPle 114
An injectable solution is prepared as
follows:
(+)-1-[[[3-(Benzoylamino)-
2-oxo-4-phenylbutyl]methyl-
amino]carbonyl~-L-proline,
sodium salt 500 g
Methyl paraben 5 g.
Propyl paraben 1 g.
Sodium chloride 25 g.
Water for injection 5
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 asceptically
filled into presterilized vials which are
closed with presterilized rubber closures. Each
vial contains 5 ml. of solution in a concentra-
tion of 100 mg. of active ingredient per ml. of
solution for injection.
In a similar manner, an injectable solution
containing 100 mg. of active ingredient per ml.
of solution can be prepared for the product of
any Examples 1 to 110.
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HA25ga
-106-
Example 115
lO00 tablets each containing the followingingredients:
~ 1-[[[3-(~oyl~no-2-oxo-4-phenylhutyl~
methy~no] OE ~ -LrProline,3-~yri-
dinylmethyl ester, hydrochloride lO0 mg.
Avicel 100 mg.
Hydrochlorothiazide 12.5 mg.
Lactose 113 mg.
Cornstarch 17.5 mg.
Stearic acid 7 mq.
350 mg.
are pxepared from sufficient bulk quantities
by slugging the t~ [[[3-(benzoylamino)-2-
lS oxo-4-phenylbutyl]methylamino]carbonyl]-L-~roline,
3-pyridinylmethyl ester,monohydrochloride, 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 is
compressed into 350 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 100 mg. of the product
of any of Examples 1 to 97 and 99 to 110.
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