Note: Descriptions are shown in the official language in which they were submitted.
:2~18f~
The present invention relate~ to new renin-
inhibiting peptides, to processes for their preparation
and to their use in medicaments, in particular in circu-
lation-influencing medicament~.
5Renin i8 a proteolytic enzyme which i~ predomi-
nantly produced by the kidneys and secreted into the
plasma. It i~ known that renin eliminates the decapeptide
angiotensin I from angiotensinogen in vivo. Angiotensin
I is in turn degraded in the lung~, the kidney~ or other
10tissues to give the octapeptide anqiotensin II, which has
an effect on blood pre~ure. The different effects of
angiotensin II such a~ ~asocon triction, Na' retention in
the Xidneys, aldosterone release in the adrenal gland and
increase in tone of ~he sympathetic nervou~ system act
15synergistically in the sense of an increa~e in blood
pressure.
The activity of the renin-angiotensin system can
be manipulated pharmacologically by the inhibition of the
activity of renin or the angiotensin converting enzyme
20(ACE) and also by blockade of angiotensin II receptors.
The development of orally administrable ACE inhibitor~
has thus led to new antihypertensives (cf. DO~ 3,628,650,
Am. J. Med. 77, 690, 1984).
A relatively new approach i8 to intervene in the
25renin-angiotensin ca~cade at an earlier point in tLme,
namely by inhibiting the highly specific peptidase renin.
Different types of renin inhibitor~ have hitherto
been developed- renin-specific antibodie~, pho~pholipids,
Le A 26 914 - 1 -
2 ~
peptides having the N-terminal sequence of prorenin,
synthe~ic peptides as substrate analogues and modified
peptides.
In EP-A2 0,273,696, EP-A~ 0,278,158 and in PCT WO
86/04~01, renin-inhibiting peptides are described in
which the scope of meaning of R2 ~see compounds according
to the invention) also includes heterocyclic radicals,
but without an indication of the specific 1,3-dithialane
or 1,3-dithione group or an actual representative of this
class of substance being given. Peptides have now been
found in which the customary amino acid radical -His
(histidine) has been replaced by a 1,3-dithiolane or 1,3-
dithione radical and which surprisingly have a good
renin-inhibiting action.
The invention relates to peptides of the general
formula (I)
~ 2 S ~ S
R1-B-tNH ~ CO)W-NH ~ O-NH ~ O-D-E-X (I)
OH
in which
R1 - represents hydrogen or
- represent~ alkoxycarbonyl having up to 8 carbon
atoms or benzylo~ycarbonyl or
- represents a grcup of the formula R3-Co-
in which
R3 - denotea morpholino or the radical~
Le A ?6~l4 - 2 -
2 ~ 8
(cH3)3c-so2-cH2-cH-
CH2-C6H5
or (CH3)3C-CO-c~2-~H-
~H2C6Hs
or
- denotes a group vf the formula -NR~R5
in which
R4 and R5 are identical or different and
denote hydxogen, straight~chain or branch-
ed alkyl having up to 8 carbon atoms or
phenyl
or
- denote the radical ~ (CH2) L-
in which
L - denotes the number O, 1 or 2
B - represents a direct bond or
- repre~ent~ an amino acid grouping of the
~ormula
R6
-NH l CO- or H3C ~ H3
in which
R8 _ denotes ~traight-chain or branched ~lkyl
having up to 8 carbon atoms, which i5
Le A 26 ~14 - 3 .
,, ., ~, . . ...
-` ~ 2 ~ 3 ~
optionally substituted by hydroxyl,
carboxyl, alkoxy or alkoxycarbonyl having
up to 6 carbon atoms, or benzyloxy
or
- represent~ proline
in its L~form, D-form or as the D,L-isomer mixture,
R2 _ represents aryl having 6 to 10 carbon atoms,
which is optionally monosubstituted to trisub~ti-
tuted by identical or different substituents from
the s~ries comprising halogen, ni~ro, cyano,
hydroxyl, traight-chain or branched alkoxy
having up to 8 carbon atoms~ acetoxy or benzyl-
oxy, or by a group of the formula o-Co-R7
in which
R7 denotes straight-chain or branched alkyl
having up to 8 ~arbon atom~
w - denotes a number 0 or 1
A - represents~a -CH~- or a CH2-CH2- group,
D and E a~e identical or dif~ere~t and
- represent a direct bond or
- repre~ent an ~mino a~id grouping of the formula
Le A 2~ 914 - 4 -
.
- ` 2 ~ 8
-NH ~ o-
in which
R6 has the abovementioned meaning of R~ and is
identical to or different from this, in its L-
form, D-form or as the D,L-isomer mixture,
X - represents hydroxyl, benzyloxy or alkoxy having
up to 8 carbon atoms or morpholino ox
- repre~ents a group of the formula -NHR8
in which
Ra _ denotes hydrogen or
- denotes ~traight-chain or branched alkyl
having up to 10 carbon atoms, which is
optionally sub~ti~uted by hydroxyl/
phenyl, pyridyl or by the radical of the
formula
~NH2
or
- repre~ent~ a radical of he formula
-- N N-CH2-C6H5
and their phy~iologically ~ceptable salt~.
The compounds of the genPral formula (I)
Le A 26 914 5
.
. ' ~
2 ~
according to ~he invention have several asymmetric carbon
atoms. ~hey can therefore be present independently of one
another in the D- or the L-form. The invention includes
the optical antipodes as well as the isomer mixtures or
racemates. PrQferably, the amino acid radicals are
present independently of one another in the optically
pure form, preferably in the L-form.
The compounds of the general formula (I) accord-
ing to the invention can be present in the form of their
salts. These may be salts of compound~ according to the
invention with inorganic or organic acids or bases. The
acid addition products preferably include salts with
hydrochloric acid, hydrobromic acid, hydroiodic acid,
sulphuric acid, phosphoric acid or with carboxylic acids
such as acetic acid, propionic acid, oxalic acid, gly-
colic acid, succinic acid, maleic acid, hydroxymaleic
acid, methylmaleic acid, fumaric acid, adipic acid, malic
acid, tartaric acid, citric acid, benzoic acid, cinnamic
acid, lactic acid, ascorbic acid, salicylic acid, 2-
acetoxybenzoic acid, nicotinic acid, isonicotinic acid,
or sulphonic ~cids such as methanesulphonic acid, ethane-
sulphonic acid, benzenesulphonic acid, toluenesulphonic
acid, naphthalene-2-sulphonic acid or naphthalene-
disulphonic acid.
Preferred compounds of thP general foxmula ~I)
are those in which
Rl ~ repre~entM hydrogen or
- repres~nts alkoxycarbonyl having up to 6 carbon
atom~ ox benzyloxycarbonyl or
Le A 26 914 - ~ -
.
~ ~'` 2~1g~
- represents a group of the formula R3-Co
in which
R3 - denotes morpholino or the radicals
( CH3 ) 3c-so2-cH2-cH-
CH2-C6H5
or (CH3)3C-CO-CH2-~H-
C~l2C~iH5
or
- denotes a group of the formula -NR4R5
in which
R4 and R5 are identical or different and
denote hydrogen, straight-chain or
branched alkyl having up to 6 carbon atoms
or phenyl
or
N
- denotes the radical ~ (CH2~L-
in which
~ - denotes the number 0, 1 or 2
Le A 26 914 - 7 -
.:
..
.
. . .
~ `` 2 ~
B - represents a direct bond or
- represents an amino acid grouping of the formula
R6
-NH ~ O-
in which
R6 _ denotes ~traight-chain or branched alkyl
having up to 6 carbon atoms, which is
optionally substituted by hydroxyl, alkoxy
having up to 4 carbon atoms or benzyloxy
ox
- represents proline,
in its L-form, D-form or as the D,L-isomer mixture,
R2 _ represents phenyl or naphthyl, which is
optionally monosubstituted to trisubstituted by
identical or different substituents from the
series compri.sing fluorine, chlorine, bramlne, jodide,
hydroxyl, nitro, cyano, straight-chain or
branched alkoxy having up to 6 carbon a~oms,
acetoxy or benzyloxy,
w denotes a number 0 or 1,
A - repre~ents a -C~z~ or a -CH2-C~2- group,
20 D and E are identical or di~ferent and
- repr~sent a direct ~ond or
- repre~en~ an amino acid grouping of the formula
Le A 26 914 - 8 -
2 ~ 8
-NH ~ o-
in which
R~ ha~ the abovementioned meaning of R5 and is
identical to or different from this, in its L-
form, ~-form or as the D,L-isomer mixture,
X - represents hydroxyl, benzyloxy or alkoxy having
up to 6 carbon atoms or morpholino or
- reRres~nts a group of the formula -NHR~
in which
R~ - denotes hydrogen or straight-chain or
branched alkyl having up to 8 carbon
atoms, which is optionally substituted by
hydroxyl, phenyl, pyridyl or by the
radical of the formula
~NH 2
or
- repres~nt~ a radical of the formula
~--~N-cH2-c6Hs
and their phy~iologically acoeptable 5alt8.
Particularly preferred compounds of the general
~ormula (I) are tho~e in which
Le A ?6 ~14 9
.
. . .
; .
`~ ~
`,' `~
^- 2 ~
R1 - represents hydrogen or
- repre~ents alkoxycarbonyl having up to 4 carbon
atoms or benzyloxycarbonyl
- represents a group of the formula R3-Co-
in which
R3 ~ denotes mDrpholino or the radical (CH3)3C-S02-CH2-~H-
or a group of the fonm~a -NR4R5 CH2 6 5
in which
R4 and R5 are identical or different and
denote hydrogen or straight-chain or
branched alkyl having up to 4 carbon
atoms, or phenyl
or
- denote~ the radical ~ (CH2)L-
lS in which
L - denotes the number 0, 1 or 2
B - repressnts a direct bond or
- xepresents an aMino acid grouping of the formula
H ~
-N 0-
in whlch
Le A 26 914 - 10
2 ~
R6 _ denotes straight-chain or branched alkyl
having up to 4 carbon atoms, which is
optionally substituted by hydroxyl
or
S - represents proline,
in its L-form, D-form or as the D,L~isomer mixture,
R2 _ represents phenyl or naphthyl, which is optionally nonosubsti-
tuted to trisubstituted by f~uorine, chlorme, iodide, hydro~l,
methoxy, ethoxy, propoxy, tert. butoxy, ~enzyloxy or acetoxy,
w - denote~ a number 0 or l,
A - represents a -CH2- or a -CH2-CH2- group,
D and E are identical or different and
- represent a direct bond or
- represent an amino acid grouping of the formula
R6 '
-NH ~ o-
in which
R6 ha~ the abovementioned meaning of R6 and is
identical to or different from thi~, in itS L-
form, D-form or as the D,L-i~omer mixture,
X - represent~ hydro~yl, benzyloxy or a}koxy having
up to 4 carbon atom~ ox morpholi~o or
- reprasent~ a group of the formula -N~R8
- 11 -
.
:
. ..
2 ~
in which
R~ - denotes hydrogen or
- denotes straight-chain or branched alkyl
having up to 6 carbon atoms, which is
optionally substituted by hydroxyl,
phenyl, pyridyl or by the radical of the
formula
~NH2
or
- represents a radical of the formula
r--~N-cH2-c6Hs
and their pnysiologically acceptable salts.
Salts of the compounds according to the invention
with salt-forming groups can be prepared in a manner
known per se, for example by reactinq the compounds
according to the invention, which contain acidic groups,
with corresponding ba~eR or by reacting the compounds
according to the invention, which contain basic groups,
with corre~ponding acid~, in each case preferably with
the abov~mentioned base~ or acid-~
Stereoisomer ~ixtureg, in particular diastereomer
mixtures, can be ~eparated .into the individual isom~rs in
a manner known per se, or example by fractional cry~tal-
lization or chromatography.
Racemates can be resolved in a manner known per
se, for exampl~ by converting the optic~l ~ntipod~s to
Le A 2~ 12 -
'
`~ 2 ~ 8
diastereomers.
The group of the formula
A~
-NH O-NH ~ O-
4 1
has three asymmetric carbon a~oms which can be present
independently of one another in the R- or S-configura-
tion. Preferably, this grouping is present in the SR, 3S,
4S-csnfiguration, 5R, 3R, 4S-configuration, 5S, 3S, 4S-
configuration or 5S, 3R, 4S-configuration, particularly
preferably in the 5R, 3S, 4S- or 5S, 3S, 4S-configu-
ration. The grouping is also employed as the isomer
mixture.
The compounds according to the invention of the
general formula (I~
A ~
~R2
R 1 - B - ( NHlCO ) W- NH~O - NH~O - D - E- X ( I )
in which OH
B, D, E, A, R~, R2, w and X have the abovementioned
meanings,
are obtain~d by a process in which
[A] compounds o the gener~l formula ~II)
Le A_26 914 - 13 -
;
~ ~ \
2 ~ 8
~ (IIJ
Z-NH ~ 0-D-E-X
in which
D, E and X have the abovementioned meanings
and
Z - represents an amino protecting group,
are first converted into amines by removing the group Z
by customary methods and these ~mines are then reacted
with compounds of the general formula (III~
S~S
Z ~ -NH~OOH t I I I ),
in which
A has the abovementioned meaning,
and
~' ha the abo~ementioned meaning of Z and i~ identical
to or different from this~
with activation of the carboxylic acid by customary
1~ methods in inert solvents to give compounds o~ the
general f~mula (Ia)
Le A 26 914 ~ 14 -
:
2 ~
A~
S ~ S ~ (Ia)
Z'-NH ~ ~-NH ~ O-D-E-X
in which OH
Z', A, D, E and X fiave the abovementioned meanings,
the protecting group Z' is removed by customary methods
and in a further step the compounds are condensed with
compounds of ~he general formula (IV)
~ 2
R1_B_(NH ~ OOH)w (IV~
in which
R1, B, w and R2 have the abovementioned meanings,
if appropriate with the abovemantioned carboxylic acid
activation,
and, if appropriate, the corresponding esters are hydro-
lyzed by customary method~, or the compounds of the
general formulae (III) and ~IV) are first reacted by the
abovementioned methods and a further peptide linX is then
at~achad using the compounds of the g-ner~1 fonmula (II3
or by a process in ~hich
l~_a_3k_2~ - 15
;, , , . , , ~ ~
,;
.: ; . ~
'
2~d-~8
~B] compounds of the general formula (Ib)
2 1 ~ S
R1-B-(NH ~ o)w-NH~A~co-NH ~ o-D-E-G
in which OH
A, B, Rl, R2, w, D and E have the abovementioned meanings,
S and
G - represents alkoxycarbonyl having up to 8 carbon
atoms,
are first hydrolyzed ~o the corresponding acids by
customary method~ and in a further ~tep are condensed in
the presence of auxiliaries with amines of the general
fo~mula (V), or morpholine of the formula (VI) or N-
benzylpiperazine of the formula (VII)
H2N-R8 ~Y), O NH (VI) or HN~__~N-CH2C6H5 (VII)
in which
R~ has the abovementioned meaning,
or by a process in which
[C] either compounds of the general formula (Ic)
Le A 26 914 - 16 -
- `" 2~31
~2 Sxs ~ ( Ic )
Rl -B- ( NH O ) -NH CO-NH~(:OX '
in which
OH
A, B, Rl, w and R2 have the abovementioned meanings,
and
X' - represents alkoxy having up to 6 carbon atoms or
benzyloxy,
or compounds of the general formula (Id)
7~ ~
s~s ,~
Z'-NH ~ O-NH ~ OX~ (Id)
in which OH
Z', A and X' have the abovementioned meanings,
are first hydrolyzed to the corresponding acids by
customary method~ and then reacted with the fragment of
~he general formula tYIII)
D-E-X ~VIII)
in which
Le ~ ?6 914 - 17 -
- ~
'. - ~
.
.
- ~ "
2~ 8~
D, E and X have the abovementioned meanings,
in inert solvents, if appropriate in the presence of
auxiliaries,
and in the case of the compounds of the general formula
5 (Id) are reacted in a next step by the method described
under process [A], with stepwise removal of the respec-
tive protecting group Z~, with compounds of the general
formula (IV) or (IVa)
~ 2 ~R2
R1_NH ~ OOH (IY) or R 1 - B - ( NH ~ OOH)W (IVa)
in which
R1, B, w and R2 have the abovementioned meanin~s.
The synthesis can be illustrated by way of
example by the following reaction scheme:
a@_~_2~_YL~ - 18
2~1g~8
~o x`
Boc-NH ~ O-NH O-NH
1. ¦ t HCl
¦ - Boc
H2N ~ O-NH ~ O-NH
~1
S~S
2. I Boc-NH~A~COOH
S~S ~ ~
Bo~~NH ~ O--NH ~ O-NH- -~O-NH
OH
Le_A 2~_~14 - l9 -
.
.
. ~ , '~ .
2~8~g
t HCl
B o c
S~XS
H2N CO--NH~Co NH--~Co- NH~3
4, ¦ ~ Boc-NH OOH
~3 5 S ~O
B o c - N O--NHXO--NH~ONHXONH~3
~A~
S S
E~OC-N OOH ~ H2NX~2H5
~ SXS
E~OC -N O-N OOC2H5
Le A 26 914 - 20 -
.
, '~ ,
,
2 ~ 8
H20 t OH
~ Sxs
EtOC-NH O~NH OOH
O '
t H2N~Co-N~l~G-NH-CH2~3
OH
~SX
EtOC-NH CO NH ~NH~CO-NH o-NH-CH2~3
OH
~B~ -
H~ ~ 1
EtOC-N X o-NHlCOOC2H5
0~
~ ~ Ol~e/H20
Le A 26 914 - 21
.. ~ .
.~ , . ,, . ~ ,
."
: . , :.
~8
,~3 r~
EtOC-NH CO- -NH X O-NH ~ COOH
OH
H2N~/\NH2 ¦-~2
~srx ~ ~
EtOC-NH CO----NH O~NH ~ ~ ONH ~ NH2
tC]
~ 3 rS ~
E~OC-NH O ~ NH X O-NH ~ OOH
OH
NH2 - CH2
OC~3
E~oc-N o HXO N~CONH-CH2
o}~
L~ A 26 ~14 - 22 -
'.
: ,
The preparation of the compounds according to the
invention has also been carried out by other customary
variants of the process described (cf. for example
Houben-Weyls ~Methoden der organischen Chemie" (Methods
of organic chemistry) XV/l and 2; M. Bodan~zky,
A. Bodan~zky in ~The Practice of Peptide Synthesis",
Springer Verlag, Berlin, 1984; George R. Pettit in
"Synthetic Peptides", Volume 4, El~evier Scientific
Publishing Company, Amsterdam-Oxford-New York, 1976;
E. Gross and J. Meienhofer (Editors) in "The Peptidesn,
Vol. 1-3, Academic Press. New York-London-Toronto-Sydney-
San Francisco, 1981; M. Bodanszky in "Principles of
Peptide Synthe6is", Springer Verlag, Berlin-Heidelberg-
New ~ork-Tokyo, 1984; R. Uhmann and K. Radscheit, Offen-
lS legung~schrift , DE 3,411,244 Al) or also by the ~Solid
Phase Method", as is described by M. Bodanszky,
A. Bodanszky in ~'The Practice of Peptide Synthesis~,
Springer Verlag, Berlin, 1984, or G. Barany,
R.B. Merrifield in ~'Solid PhasQ Peptide Synthesis~ from
"The Peptide~", Vol. 2, p. 3-254, edited by E. Gross,
J. Meienhofer, Academic Pres~, New York-London-Toronto~
Sydney-San Francisco ~1980).
Suitable ~olvents in process variant~ [A], [B]
and [C] are the customary inert solvent~ which do not
change under the reaction conditions selected in esch
case. These include water or organic 801vent8 ~uch a~
diethyl ether, glycol monomethyl ether or glycol dLme~hyl
ether, dioxane or tetrahydrofuran, or hydrocarbon~ ~uch
as benzan~, toluene, xylene, cyclohexane or mineral oil
fraction~ or halogenated hydrocarbon~ such as methylene
Le A 26~14 - 23 -
. , .
~3~5~
chloride, chloroform, carbon tetrachloride or acetone,
dLmethyl sulphoxide, dimethylformamide, hexamethyl-
phosphoramide, ethyl acetate, pyridine, triethylamine or
picoline.
It .is likewise possible to use mixtures of the
solvents mentioned.
Tetrahydrofuran, methylene chloride, dimethyl-
formamide an~ ethyl acetate are preferred.
The process variants ~A~, [B] and tC] are custo-
marily carried out in ~he presence of suitable solvents
or diluents, if appropriate in the presence of an auxi-
liary or catalyst in a temperature range from -80C to
300C, preferably from -30C to +30C at normal pressure.
It is likewise possible to work at elevated or reduced
pressure.
Suitable activated carboxyl groups in process
variants [A]~ [B] and [C] are, for example, carboxylic
acid azides (obtainable, for exampl~, by reaction of
protected or unprotected carboxylic acid hydrazides with
nitrous acid, its salts or ~lkyl nitrites (for example
i~oamyl nitrite),
or unsaturated esters, in particular vinyl esters,
(obtainable, for example, by reaction of a corresponding
ester with ~inyl acetate), carbamoylvinyl esters
(obtainable, for example, by reaction of A corresponding
acid with an isoxazolium reagent), alkoxyvinyl ~sters
(obtainable~ ~or example, by reaction of the correspond-
ing acid~ with alkoxyacetylenes, pref~rably ethoxy~
acetylene),
or amidino ~8ter8, for exa~ple, N,N'- or N,N-
Le A 26 914 - 24 -
2 ~ 8
disubstituted amidino esters (~btainable, for example, by
reaction of the corresponding acid with an N,N'-disub-
stituted carbodiimide (preferably di~yclohexylcarbodi-
Lmide, diisopropylcarbodiimide or N-(3-dimethylamino-
propyl)-N'-ethylcarbodiLmide hydrochloride) or with an
N,N-disubstituted cyanamide,
or a~yl esters, in particular by phenyl esters substi-
tuted with electron-withdrawing ~ubstituent~, for example
4-nitrophenyl, 4-methylsulphonylphenyl, 2,4 t S-trichloro-
phenyl, 2,3,4,5,6-pentachlorophenyl or 4-phenyldiazo-
phenyl esters (obtainable, for example, by reaction of
the corresponding acid with an appropriately ~ubstituted
phenol, if appropriate in the presence of a condensing
agent such as, for example, N,N~-dicycloh~xylcarbo-
diimide, diisopropylcarbodiLmide, N~(3-dLmethylamino-
propyl~-N'-ethylcarbodiimide hydrochloride, i~obutyl
chloroformate, propanephosphonic anhydride or benzotria-
zolyloxytris(dimethylamino)phosphonium hexafluorophos-
phate)~
or cyanomethyl ester~ (obtainable, for example, by
reaction of the corresponding acid with chloroaceto-
nitrile in the presence of a ba~e),
or thioesters, in particular nitrophenylthioesters
(obtainable, for e~mple, by reaction of the correspond-
ing acid with nitrothiophenol~, if appropri~te in thepresence of conden in~ agents such as N,N' dicyclohexyl-
carbodiimide, diisopropylc~rbodiimide, ~-(3-dimethyl-
aminopropyl)-N'-ethylcarbodilmidehydro~hloride,i~obutyl
chloroformate, propanephosphonic anhydride or
benzotriazolyloxy_tri ~dimethylamino)phosphonium
Le A 2~ 914 - 25 -
'; .
2V~8~g
hexafluorophosphate),
or amino or amido esters (obtainable, for example, by
reaction of the corre~ponding acid with an N-hydroxyamino
or N-hydroxya~ido compound, in particular N-hydroxy-
succinimide, N-hydroxypiperidine, N-hydroxyphthalLmide,
N-hydroxy-5-norbornene-2,3-dicarboxLmide or l-hydroxy-
benzotriazole, if appropriate in the presence of condens-
ing agents ~uch as N,N'-dicyclohexylcarbodiLmide, diiso-
propylcarb~diimide or N-(3-dimethylaminopropyl)-N'-
ethylcarbodiimide hydrochloride, isobutyl chloroformateor n-propanepho~phonic acid anhydride),
or anhydrides of acids, preferably symmetrical or unsym-
metrical anhydrides of the corresponding acids, in
particular anhydrides with inorganic acids, (obtainable,
for example, by reaction of the corresponding acid with
thionyl chloride, phosphorus pentoxide or oxalyl chlo-
ride), or anhydride~ with carbonic acid half derivatives,
for example, carbonic acid lower alkyl half esters
(obtainable, for example, by reaction of the correspond-
ing acid with halogenoformic acid lower alkyl esters, forexample methyl chloroformate, ethyl chloroformate, propyl
chloroformate, isopropyl chloroformate, butyl chlorofor-
mate or isobutyl chloroforma~e or with 1-lower alkoxy-
carbonyl-2-lower alkoxy-1,2-dihydro-quinoline, for
example 1-methsxycarbonyl-2-ethoxy-1,2-dihydroquinoline),
or anhydrides with dihalogenophosphoric acid~ (obtain-
able, for ex~mple, by reaction of ~he corr~sponding acid
with phosphoru~ oxychloridel,
or anhydrides with phosphori~ acid derivatives or pho~-
phorous ac~d deriva~ives, ~for example propanephosphonic
Le A 26 914 - 26 -
2 ~
anhydride, H. Wissmann and H.J. Kleiner, Angew. Chem.
Int. Ed. 19, 133 (1980))
or anhydrides with organic carboxylic acids (obtainable,
for example, by reaction of the corresponding acids with
S an optionally substi~uted lower alkane- or phenylalkane-
carboxyl halide, in particular phenylacetyl, pivaloyl or
trifluoroacetyl chloride),
or anhydrides with organic sulphonic acid~ (obtainable~
for example, by reaction of an alkali metal salt of a
corresponding acid with a sulphonyl halide, in particular
methane-, ethane-, benzene- or toluenesulphonyl chlo-
ride),
or symmetrical anhydrides (obtainable, for example, by
condensation of corresponding acids, if appropriate in
the presence of condensing agents such as N,N'-dicyclo-
hexylcarbodiimide, dii~opropylcarbodiimide, N-~3-di-
methylaminopropyl)-N'-ethylcarbodiimide hydrochloride,
isobutyl chloroformate, propanephosphonic anhydride or
benzotriazolyloxy-~ris(dimethylamino)phosphonium hexa-
fluorophosphate).
Reactive cyclic amides are, in particular, amideswith five-membered heterocycles haYing 2 nitroqen atoms
and optionally aromatic character, preferably amides with
imidazoles or pyrazole6 (obt~inable, for example, by
reaction of the corresponding acids with N,N'-carbonyl-
diimidazole or - if appropriate in the presence of
condensing agents ~uch as, for example, N,N'-dicyclo-
hexylcarbodiimide, N,N'-diisopropylcarbodiimide, N-(3-
dimethyl~minopropyl)-N'-ethylcarbodiimide hydrochloride,
isobutyl chloroformate, propanephosphonic anhy~ride,
Le A~ 2~ 914 - 27 -
:
2 ~ 8
benzotriazolyloxy-tris(dimethylamino)phosphonium hexa-
fluorophosphate - with, for example, 3,5-dLmethyl-
pyrazole, 1,2,4-triazole or tetrazole.
Suitable amino protecting groups are the amino
protecting groups customa~y in peptide chemistry.
These preferably includes benzyloxycarbonyl, 4-
bromobenzyloxycarbonyl, 2-chlorobenzyloxycarbonyl, 3-
chlorobenzyloxycarbonyl, dichlorobenzylo~ycarbonyl, 3,4-
dimethoxyb~nzyloxycarbonyl, 3,5-dimethoxybenzyloxy-
carbonyl, 2,4-dLm~thoxyben2yloxycarbonyl, 4-methoxy-
benzyloxycarbonyl, 4-nitrobenzyloxycarbonyl, 2-nitro-
benzyloxycarbonyl, 2-nitro-4,5-dimethoxybenzyloxy-
carbonyl, 3,4,5-trimethoxybenzyloxycarbonyl, methoxy-
carbonyl, ethoxycarbonyl, prop~xycarbonyl, isopropoxy-
carbonyl, butoxycarbonyl, isobutoxycarbonyl, tert-butoxy-
carbonyl, pentoxycarbonyl, isopentoxycarbonyl, hexoxy-
carbonyl, cyclohexoxycarbonyl, octoxycarbonyl, 2-ethyl-
hexo~ycarbonyl, ~-iodohexoxycarbonyl, 2-bromoethoxy-
carbonyll 2-chloroethoxycarbonyl, 2,2,2-trichloroethoxy-
carbonyl, 2,2,2-trichloro-tert-butoxycarbonyl, benz-
hydryloxycarbonyl, bis-(4-methoxyphenyl)methoxycarbonyl,
phenacyloxycarbonyl, 2-trimethylsilylethoxycarbonyl, 2-
(di-n-butyl-methylsilyl)ethoxycarbonyl, 2-triphenyl-
silylethoxycarbonyl), 2-(dimethyl-tert-butylsilyl)etho~y-
carbonyl, menthyloxycarbonyl, vinyloxycarbonyl, allyloxy-
carbonyl, phenoxycarbonyl, tolyloxycarbonyl, 2,4-dinitro-
phenoxycarbonyl,4-nitrophenoxycarbonyl,2,4,5-trichloro-
pheno~ycarbonyl, naphthyloxycarbonyl, fluorenyl-9-
methoxycarbonyl, ethylthiocarbonyl~ methylthiocarbonyl,
butylth$ocarbonyl, tert.-bu~ylthiocarbonyl,
L~ A 26 914 - 28 -
2 ~
phenylthiocarbonyl, benzylthiocarbonyl, methylamino-
carbonyl, e~hylaminocarbonyl, methylthiocarbonyl, butyl-
thiocarbonyl,tert-butylthiocarbonyl,phenylthiocarbonyl,
benzylthiocarbonyl, methylaminocarbonyl, ethylamino-
carbonyl, propylaminocarbonyl, iso-propylaminocarbonyl,
formyl, acetyl, propionyl, pivaloyl, 2-chloroacetyl, 2-
bromoacetyl, 2-iodoacetyl, 2,2,2-trifluoroacetyl, 2,2,2-
trichloroacetyl, benzoyl, 4-chlorobenzoyl, 4-methoxy-
benzoyl, 4rnitrobenzyl, 4-nitrobenzoyl, naphthylcarbonyl,
phenoxyacetyl,adamantylcarbonyl,dicyclohexylphosphoryl,
diphenylphosphoryl, dibenzylphosphoryl, di-(4-nitro-
benzyl)phosphoryl, phenoxyphenylphosphoryl, diethylphos-
phinyl, diphenylphosphinyl, phthaloyl or phthalLmido.
Particularly preferred amino protecting groups
are benzyloxycarbonyl, 3,4-dLmethoxybenzyloxycar~onyl,
3,5-dimethoxybenzyloxycarbonyl, 4-m~thoxybenzyloxy-
carbonyl, 4-nitrobenzyloxycarbonyl, 2-nitrobenzyloxy-
carbonyl, 3,4,5-trLmethoxybenzyloxycarbonyl, methoxy-
carbonyl, ethoxycarbonyl, propoxycarbonyl, isopropoxy-
carbonyl, butoxycarbonyl, isobutoxycarbonyl, tert-butoxy-
carbonyl, cyclohexoxycarbonyl, hexoxycarbonyl, octoxy-
carbonyl, 2-bromoethoxycarbonyl, 2-chloroethoxycarbonyl,
phenoxyacetyl, naphthylcarbonyl, adamantylcarbonyl,
phthaloyl, ~,2,2-trichloroethoxycarbonyl, 2,2,2-
trichloro-tert-butoxycarbonyl, menthyloxycarbonyl,
~inyloxyoarbonyl, allyloxycarbonyl, phenoxycaxbonyl, 4-
nitrophenoxycarbonyl, fluorenyl-9-methoxycarbonyl,
formyl, acetyl, propionyl, pivaloyl, 2 hloroacetyl, 2-
bromoacetyl, 2,2,2-trifluoroacetyl, 2,2,2-trichloro-
acetyl, benzoyl, 4-chlorobenzoyl, 4-br~mob~nzoyl,
Le A 26 914 - 29 -
2 ~
4-nitrobenzoyl, phthalimido or isovaleroyl [cf. Th. W.
Greene, Protective Groups in Organic Synthesis, John
Wiley & Sons, New York 1981].
Auxiliaries employed are preferably condensing
agents, which may also be bases, in particular if the
carboxyl group is activated as the anhydride. ~referably,
the customary condensing agents such as carbodiimide, for
example N,N'-diethyl-, N,N'~dipropyl-, N,N'-dii~opropyl-
or N,N'-dicyclohexylcarbodiimid~, N-(3-dimethylamino-
isopropyl)-N~-ethylcarbodiimide hydrochloride, or car-
bonyl compounds such a~ car~onyldiimidazole, or 1,2-
oxazolium compounds such as 2-ethyl-5-phenyl-1,2-oxa-
zolium 3-sulphate or 2-tert-butyl-5-methyl-oxazolium
perchlorate, or acylamino compounds such as 2-ethoxy-1-
lS ethoxycarbonyl-1,2-dihydroquinoline,orpropanephosphonic
anhydride, or isobutyl chloroformate, or benzotriazolyl-
oxy-tris(dLmethylamino)pho~phonium hexafluorophosphate
and, as bases, alkali metal carbonates, for example
sodium carbonate or pota~sium carbonate or sodium hydro-
gen carbonate or potassium hydrogen carbonate, or organic
bases such as trialkylamines, for example triethylamine,
N-ethylmorphol$ne, N-m~thylpiperidine or diisopropyl-
ethylam$ne are employed here.
The removal of the amino protecting group is
carried out in a manner known per ~e under acidic or
basic condition~, or reductively by catalytic hydrogena-
tion, for example, with Pd/C in or~anic solvent3 ~uch as
ether~, for example totrahydrofuran or dioxane, or
alcohol~, for example methanol~ eth~nol or isQpropanol.
The hydroly~i~ of the carboxylic ac~d esters i8
ke A ?6 914 _ 30 _
! ` `
'
,
2 ~
carried out by customary methods in one of the above-
mentioned solvents, by treating the esters with customary
bases, it being possible to convert the salts initially
formed into the free carboxylic acids by treating with
acid.
Suitable bases for the hydrolysis are the cus-
tomary inorganic bases. These preferably include alkali
metal hydroxides or alkaline earth metal hydroxides such
as, for example, sodium hydroxide, potassium hydroxide or
barium hydroxide, or alkali metal carbonates such as
sodium carbonate ox pota~sium carbonate or ~odium hydro-
gen carbonate, or alkali metal alkoxides such as sodium
ethoxide, sodium methoxide, pota~sium ethoxide, potassium
methoxide or potassium tert.bu~oxide. Sodium hydroxide or
potacsium hydroxide are particularly preferably employed.
The hydroly~is i~ in general carried out in a
temperature range from 0C to +100C, preferably from
~20C to ~80~.
In general, the hydrolysi~ is carried out at
normal pressure. However, it is also possibla to work at
reduced pre~sure or at elevated pressure (for example
from 0.5 to 5 bar).
When carrying out the hydrolysis, the base i8 in
general employed in an amount of from 1 ~o 3 moles,
preferably from 1 to 1.5 moles, relative to 1 mole of the
e~ter. Molar amounts of the reactan 8 are particularly
preferably u~ed.
The removal of the e~terq can likewi~e be carried
out by cu~tomary method~ using acids, ~uch a~, for
example, hydrochloric acid or trifluoroacetic acid in the
Le A 26 ~14 - 31 -
'
,
2~g~
case of the tert.-butyl esters or by hydrogenolysi~ in
the case of benzyl esters.
The compounds of the general formulae (II), (IV),
(IVa) and ~VIII) are known per se or can be synthesized
by customary methods of peptide chemistry, by reacting a
coxresponding fragment consisting of one or more amino
acid grouping~, containing a free carboxyl group, if
appropriate present in activatad form, wîth a comple-
mentary fragment, consisting of one or more amino acid
groupings, containing an amino group, if appropriate in
activated form, and optionally repeating this procedure
with appropriate fragments until the desired peptides of
the general formulae (II), (IY) and (IVa) have been
prepared, and sub~equently optionally removing protecting
groups or replacing by other protecting groups.
In this connection, additional reactive groups,
such as, for example, amino or hydroxyl groups, in the
side chains of the fragments can optionally be protected
by customary protecting groups [cf. Houben-Weyl, Eugen
Muller, Methoden der organischen Chemi~, (Methods of
organic chemistry), Volume XV/l and Volum~ XV/2, Georg
Thieme Verlag, Stuttgart, 1974].
The reaction with morpholine in (VI) and N-
ben~ylpiperazine in (VII) or with amines of the formula
(V) i~ in ~eneral carried out in one of the above-
mentioned inert solvents, in the pre~ence of one of the
abovementioned base~, preferably in triethylamine and
mcthylena chloride at a temperature of -40-C to O~C,
preferably at -20~C and normal pre~surQ.
In ~eneral, 1 to 5, pre~erably 1.5 to 1 mole, of
Le A ?6 914 - 32 -
.
.
.
1 3 ~
amine, relative to 1 mole of the reaction partner, are
employed.
The compounds of the general formula (III)
A ~
S~S
Z -N~ ~ OOH ~III),
in which
A and Z' have the abovementioned meanings,
are new and can be prepared by a process in which
compound~ of the general formula (IX)
S~S
H2N ~ 00~ ~IX),
in which
A has the abovementioned meaning,
are reacted with a reagent, such as, for example,
( Z ~ - O -CO ) 2 ~ Z ~ -O-CO-C1 or (Z'-O-CO)-O-N-succinimide,
introducing the amino protecting group Z' by the method
[A] described above in inert solve~ts in the presence of
a base, preferably with sodium hydr~xide in a dioxane~
lS water mixture or with triethylamine in dioxane, in a
temperature range from O~C to ~50C, preferably at room
temperature and nonmal pre~sure.
The compsunds o~ the general formulae ~Ia) and
(Ib~ ar~ n~ a~d ar~ prepared by the methods described
under proc~s~es ~A] nd ~B].
~he compounds of the genersl fonmula (Id) are
I.s A ?~ 914 33 ~
2 ~
also new and can be prepared by a process in which
compounds of the general formula (III)
~'
S~S
Z'-NH ~ OOH (III),
in which
Z' and A have the abovementioned meanings,
are reacted with com ~ nds of the general formula (X)
~V
ZNH ~ OX' (X),
in which OH
Z and X' have the abo~ementioned meanings,
with removal of the protecting group Z.
The compound~ of the general formula (X) are
10known ~cf. Boger et al., J. Med. Chem. 28, 1779-1790
(1985)].
The compounds of the general formula (IX) are
known or can be prepared by known methods [M.P. Mertes,
A.A. Ramsey, J. Med. Chem. I2, 342 (1969)].
15The compounds of the general formula (Ic~ are
also new and are prepared by reaction of the compounds of
the formulae (III), (IV) and (X) by the cu tomary peptide
~oupling method~ described under proce s [A].
The amines of the general formulae (V), 5VIII)
and (IX) are known (cf. G.C. Barrett, Ch~mistry of the
~mino Acids, Chapman and Hall, New York, London, 1988 and
~P-A2 0,278,158].
L~ A 26 914 - 34 _
2 ~ 1 ~ Ls~ ~ ~
Morpholine and N-benzylpiperazine are known.
The compounds according to the invention have a
circulation-influencing effect and can therefore be
employed in medicaments ~or the treatment of blood
pressure and cardiac insufficiency.
In vitro test
The inhibitory potency of the psptides accordinq
to the invention against endogenous renin from human
plasma is determined in vitro. Pooled human plasma is
obtained with the addition o ethylenediaminetetraacetic
acid tEDTA) a~ anticoagulant and stored at -20C. The
plasma renin activity ~PRA) is determined by incubation
a~ 37C as the rate of formation of angiotensin I from
endogenous angiotensinogen and renin. The reaction
solution contains 150 ~1 of plasma, 3 ~1 of 6.6~ strength
8-hydroxyquinoline sulphate solution, 3 ~1 of 10~
strength dimercaprol solution and 144 ~1 of sodium
phosphate buffer (0.2 M 0.1~ EDTA; pH 5.6) with or
without the substances according to the invention in
various concentrations. The angiotensin I ~ormed per unit
of time is determined by r~dioLmmunoassay (Sorin
Biomedica, Italy). The percentage inhibi~ion o the
plasma renin activity i8 calculated ~y ~omparison of the
substances claimed herein. ~he concentra~ion range in
which the substances claimed herein show a 50~ inhibition
of th~ plasma xenin activity i~ between 10-7 and 10-1 M.
Le A 26 914 - 35 -
~ 3
Example No. IC50 [M]
IV 1.1 . 10-7
V 7.0 . 10-1
XII 7.4 10-8
XIII 1.4 o 101
XVI 2.4 . 10
XVII 1.9 10-1
XXXII 1.5 10~
XXXIII 2.2 101
The new active compounds can be converted in a
c~stomary manner into the customary formulations, such as
tablets, coated tablets, pill5, granules, aerosols,
syrups, emulsions, suspensions and solutions, u~ing
inert, non-toxic, pharmaceutically suitable excipients or
solvents. In this connection, the therapeutically active
compound should in each case be present in a concen- -
txation from about 0.5 to 90~ by weight of tAe total
mixture, i.e. in amounts which are ~ufficient in order to
achiev~ the dosage range indicated.
The formulation& are prepared, for example, by
~xtending the active compounds with solvents and/or
excipientY, optionally using emul3ifiers and/or di~per-
~an~s, it being poYsible, for example, in the c~se of the
use of water as a diluent, to u~e, if appropriate,
organic solvents as auxiliary ~olvent~. :
ExampleR of auxiliary ~olvents which may be
mentioned ara 8
water, non-toxic org~nic solvents, 3uch a~ para~in~ (for
30 example mineral oil fraction~ ) ~ v~getabl~ oil~ (for
Le A_?6 9~14 - 36
`
example groundnut/sesame oil), alcohols (for ~xample:
ethyl alcohol, glycerol), excipients, such a~, for
example, grou~d natural mineral~ (for example kaolins,
aluminas, talc, chalk), ground ~ynthetic minerals (for
S example highly di~perse silica, silicates), sugar~ (for
example ~ucrose, lactose and dextrose), emulsifier~ (for
example polyoxyethylene fatty acid esters), polyoxy-
ethylene fatty alcohol ethers (for example lignin-
sulphite waste liquors, methyl cellulose, starch and
polyvinylpyrrolidone) and lubricants (for example mag-
nesium stearate, talc, stearic acid and sodium sulphate).
Administration is carried out in a customary
manner, preferably orally or parenterally, in particular
perlingually or intravenously. In the case of oral
administration, in addition to the excipients mentioned,
tablets may of cour6e also contain additions, such as
sodium citrate, calcium carbonate and dicalcium phosphate
together with ~arious additives, such as starch, prefer-
ably potato starch, gelatine and the like. Furthermore,
lubricants, such a~ magne~ium stearate, sodium lauryl
sulphate and talc may additionally be used for tableting.
In the case of aqueous suspension~, various flavor
enhancers or coloran~s may be added to the active com-
p~unds in addition to the abovementioned auxiliaries.
In the ca~e of parenteral admini~tration, ~olu-
tions of the active compounds u~ing 3uit~ble liquid
excipient ma~erials may be employed~
In general, it ha~ prov~d advantageous on intra-
venou~ ~dmini~tration to administer ~mounts of about
0.001 to 1 mg/kg, prefexably about 0.01 to 0.5 mg/kg of
Le A 26 ~14 - 37 -
.
body weight to attain effective results, and on oral
admini~tration the dosage is about 0.01 to 30 mg/kg,
preferably 0.1 to 10 mg/kg of body weight.
In spite of this, it may sometimes be neces~ary
S to deviate from the amounts mentioned, in particular
depending on the body weight of the experimental anLmal
or the type of administration, but also on the basi~ of
the animal species and its individual behavlor towards
the medicament or the na~ure of it~ formulation and the
point in time or interval at which administration take~
place. Thus, in some case~ it may be sufficient to manaqe
with less than the minimum amount previously mentioned,
while in other cases the upper limits mentioned must be
exceedPd. In the case of the admini6tration of relati~ely
large amount6, it may be advisable to divide these into
a number of individual doses over the day. For admini-
stration in human medicine, the same dosage range is
intended. ~ccordingly, the above embodiments also apply
in this case.
Starting compounds
Example_l (Formula VIII, amino function protected by
protecting group Boc)
N-tert.-bu~oxycarbonyl-L-isoleucine-(2-picolyl)amide
Bo -NH X ON ~
419.13 ml (3.024 mol) o~ triethyl~mine are added
at O~C to a solutlon of 100 ~ ~0.432 nol) of N-tert.-
butoxyc~rbonyl-L-isoleucine ~nd 46.72 g tO.432 mol) of
Le A 26 914 - 38 -
.
2-picolylamine in 1 1 of methylene chloride. The mixture
is tirred for 10 minute~ and cooled to -20C (dry ice/-
acetone)~ and 365 ml (0.561 mol) of a 50~ strength
solution of propanephosphonic acid anhydride m me~hylene
chloride (~oechst AG) is addad at this temperature. The
mixture is stirred further at -20C for 1 hour and
brought to room temperature overnight. The reaction
mixture i~ extracted by ~ucces~ively shaking 3 tLmes with
300 ml of 54 strength sodium hydrogen carbonate solution
each time, 3 time~ with 300 ml of pH 4 buffer solution
(Merck, Art. No. 9435) each time and finally twice with
300 ml of saturated sodium chloride solution each tLme.
The organic phase i dried over sodium sulphate and
concentra ed in vacuo. The crude produc~ is crystallized
by stirring with diethyl ether, filtered off with suction
and then recxystallized from diisopropyl ether~n-hexane.
Yield: 85.9 g (61.8~ of theory)
TLC system II : R~ = 0.35
TLC system III: R~ = 0.49
TLC system IV : Rr = 0 57
Example 2 (Formula VIII)
L-Isoleucine-(2-picolyl)amide dihydrochlorida
HCl x H2 ~ ON ~ x HCl
250 ml of 4 N hydrochloric acid (gas) in dioxane
are added to 40 g (0.124 mol) of ~he compound from
Example l and the mixture i8 ~tirred with ice cooling.
The suspension i~ m~de to di~solve by adding 10 ml of
methanol. Aft~r compl~tion of the reaction (TLC
Le A 26 914 - 39 -
2 a ~ r! ~ ~ ~
checking), the mixture is concentrated on a rotary
evaporator. After repeated coevaporation with diethyl
ether, the crude product is stirred with diethyl ether,
filtered off with suction and dried over potassium
hydroxide in a desiccator.
Yield: 37.2 g (100~ of theory)
TLC system : Rf = 0.30
Example 3 ~Formula II)
N-tert.-butoxycarbonyl-4S-amino-3S-hydroxy-5-cyclohexyl-
pentanoyl-L-isoleucine-(2-picolyl)amide
'~ ~
~oc-N ~ 0-N ~N ~
420 ml of methylene chloride are added to 13.2 g
~41.8 mmol) of N-tert.-butoxycarbonyl-4S-amino-3S-
hydroxy-cyclohexyl-pentanoic acid [J. Boger et al., J.
Med. Chem. 28, 1779 (1985)] and 13.5 g (46 mmol) of the
lS compound from Example 2. The suspension i8 made to
dissolve by adding 31 ml (230 mmol) of triethylamine and
is cooled to -20C (acetone/dry ice). 39 ml of a 50
strength sDlution ~f pr~ephosphonic acid anhydride m
methylene chloride are added dropwise with 6tirring at
this temperature and the mixture i~ allowed to come to
room temperature overnight. The methylene chloride i8
stripped off on a rotary evaporator and the residue i8
taken up In 400 ml of ethyl acetate. The organic phase i~
succ~i~ely washed once with 200 ml of ~aturated sodium
hydrogencarbonate solution, twice with 200 ml of buffer
Le A ~6 914 - 40 -
.
solution pH 7 (Merck, Art. No. 9439) each tLme and 4
times with 200 ml of demineralized water each tLme. The
buffer solutions are back-extracted using 100 ml of ethyl
acetate each. The combined organic phases are washed
twice using 500 ml of saturated sodium chloride solution
each time, dried over ~odium sulphate and concentrated on
a rotary evaporator to a volumQ of about 125 ml. 125 ml
of n-hexane i8 added to the solution, which is placed in
a refriqerator (5C) overnight. The product which
crystallizes out is fil~ered off with suction, washed
with a little cold athyl acetate~n-hexane 1:1 and dried
in vacuo.
Yield : 17.2 g (79% of theory)
MS-DCI: m/e 519 (M~H)
TLC system III: Rf = 0.48
TLC system IV : Rf = O . 63
TLC sy~tem V : R~ = 0.48
HPLC system II: ~ = 6.02 min
Example 4 (Formula II)
4S-amino-3S-hydroxy-5-cyclohexyl-pentanoyl-L-i~oleucine-
(2-picolyl)amide dihydrochloride
OH ~ x HCl
160 ml of 4N hydrochlori~ acid (ga8) in dioxane
are added wi~h ice cool~ng to 16 g (30.8 mmol) of the
compound from ~xa~ple 3. The ~u8p*n~ion is made to di~-
~olve ~y adding 5 ml of methanol. ~fter c~mpletion of ~he
Le A 26 914 - 41 -
~ 3
reaction (TLC checking), the mixture is concentrated on
the rotary evaporator. ~fter repeated coevaporation with
diethyl ether, the crude product is stirred with diethyl
ether, filtered off with suction and dried over sodium
hydroxide in a desiccator.
Yield: 16.56 g (95~ of theory)
MS-DCI: m/e 419 (M+H)
TLC ystem III : Rr = 0 35
Example 5 (Formula III)
N-tert.-bu~oxycarbonyl-2-R,S-amino-2-[2-(1,3-dithio-
lano)]acetic acid
r~
S~S
Bo -NH ~ O2H
R,S
19.9 g (0.11 mol) of 2-amino-2-R,S~2-[2-(1,3-
dithiolano)]acetic acid [M.P. Mertes, A.A. Ramsey, J.
Med. Chem. 12, 342 (1969)] are dissolved in 100 ml of
dioxane and 6.6 g (0.16 mol) of sodium hydroxide and 29 g
(0.13 mol) of di-tert.-butyl carbonate are added. The
mixture is stixred at room temperature overnight and the
dioxane is stripped off on a rotary evaporator. The
mixture i~ made up with 200 ml of water and extracted
several times with ether. The ba~ic 8queou~ phase i6
ad~u~ted to pH 3 with hydrochloric acid and extracted
s~veral times with methylene chloride. The combined
organic pha~es are wash~d with ~aturated odium chloride
solutlon, dried over ~odium sulphate and concentrated.
Yields 18.1 g (60.6% of theory~
MS-DCIs m/e 280 (M~H); m/e 297 ( ~NH4 )
Le A 26 914 - 42 -
TLC system IV : Rf = O . 55
Example 6 (Formula II)
N-tert.-~utoxycarbonyl-4S-amino-3S-hydro~y-5-cyclohexyl-
pentanoyl-L-leucine ethyl ester
Boc-NH ~ O-N C2H5
0~
The title compound i8 obtained analogously to
E~mple 3 by pro~anephosphonic acid anhvdride coupling of
4.8 g t15.2 mmol) of N-tert.-butoxycarbonyl-4S-amino-3S-
hydroxy-5-cyclohexylpentanoic acid lJ. Boger et al., J.
Med. Chem. 28, 1779 (198S)] and 3.3 g (15.7 mmol) of L-
leucine ethyl ester hydrochloride.
Yield: 6.4 g (92% of theory)
TLC ~ys~em VI : R~ = 0.85
TLC system VII : Rf = 0.13
TLC system VIII: Rf = O . 55
TLC system IX : Rf = O . 24
Example 7 (Formula II)
4S-amino-3S-hydroxy-5-cyclohexylpentanoyl~L-leucine ethyl
ester hydrochloride
¦ H ¦
H ~ C
HCl x NH2 ~ O-N OOCH2CH~
OH
The tltle compound i~ obtained analogously to
Examplo 4 by removlng the N-tert.-butoxycarbonyl protect-
ing group ~rom 6.1 g (13.3 ~molJ of the compound from
Le A 26 914 - 43 -
Example 6.
Yield: 5.1 g (98% of theory)
(+) FAB-MS: m/e 357 (M+H); m~e 379 (M+Na).
TLC system III : Rf = 0.37
TLC system IV : Rf = 0 . 29
TLC system VI : R~ = 0.85
TLC system VII : R~ = 0.13
TLC system VIII: R~ = 0.55
TLC system IX : Rf = 0.24
Example 8
N-tert.-butoxycarbonyl-4S-amino-3S-hydroxy-5-cyclohexyl-
pentanoyl-(2S-methyl)butylamide
,~3
Boc -NH~ONllr
OH
The title compound i8 o~tained analogously to
E~le 3 by propanephosphonic acid anhydride couplmg of
4.8 g (15.2 mmol) of N-tert.-butoxycarbonyl-4S-amino-3S-
hydroxy-5-cyclohexylpen~anoic acid [J. Boger et al., J.
Med. Chem. 28, 1779 (1985)] and 1.9 ml (16.7 mmol) of ~2-
S-methyl)butylamine.
Yield: 5.4 g (92.4% of theory)
TLC system IV s R~ = 0.72
HLPC system II : ~ = 13.51 min
(~3 FAB-~S s m/e 385 (~+H); m/e 391 (~Li)
Ex3mple 9
4S-~mino-3S-hydroxy-5-cyclohexylpentanoyl-~2S-methyl)-
butylamide hydrochloride
Le A 26 914 _ 44 _
2 ~ ~J
HC1 x N~2 ~ ON~
OH
The title compound i~ obtained analogously to
Example 4 by removal of the Boc protecting group from
5.4 g (14 mmol) of tha compound ~rom Example 8.
Yield: 5 g (~ 100% of theory, HCl-containing)
S TLC system III : R~ - Q.26
(+)FAB-NS : m/e 285 (M+H)
ExampIe 10
Ethyl N-phthaloyl-2-R,S-amino-2-12-(1,3-dithiano~]acetate
~ N 1~ C2Hs
The title compound is prepared in the manner
described in the literature for the corre~pondinq dithio-
lano compound [M.P. Mertes, A.A. Ramsey, J. Med. Chem.
I2, 342 (1969)] starting from 77.8 g (0.3 mol~ of ~-
phthalimidomalonaldehyde ethyl e~ter and 90 ml ~0.9 mol)
of 1,3-dimercaptopropane. The crude product i8 purified
by silica qel chromatography using dichloromethane as the
eluent.
Yield: 74 ~ (~0~ of theory)
TLC ~y~te~ II s R~ = O.89
TLC sy~te~ XI : F~ = 0.44
HPLC systQm II: R~ 2 6.63 min
Le A 26 ~I4 - 45
~g~8
MS m/e 352 (M~H); mJe 204; m/e 119
H-NMR (250 MHz, DMSO): ~: 7.92 (m, 4H3; 5.25 (d, lH);
4.62 (d, lH); 4,15 (~, 2~);
3.03 (m~ lH); 2.72 (m, 2H);
2.57 (m, lH); 1.95 (m, lH);
1.78 (m, lH); 1.14 (t, 3H);
Example 11
Ethyl 2-R~S-amino-2-[2-(1,3-dithianO)]acetate
S~S
NH2~cooc2H5
R,S
The title compound is prepared in the manner
described in the literature for the corresponding dithio-
lane compound [M.P. Mertes, A.A. Ramsey, J. Med. Chem.
12, 34~ (1969)] starting from 74 g (0.21 mol) of the
compound from Example 10 and 17 ml ~0.35 mol) of hydra-
zine hydrate.
Yield: 30 g (64% o~ theory)
TLC sys~em II s R~ = 0.30
(+) FAB-MS : m/e 222 (M+H) m~e 228 (~Li); m~e 244
(M~Na).
lH-NMR ~250 NHz, DMSO): ~s 4.10 (m, 3H); 3.65 (d, lH3;
2.80 (m~ 4H); 1.92 (m, 3H);
1.78 (m, lH); 1.18 (t, 3H).
Example 12
2-R,S-a~ino-2-[2-(1,3-dithiano)acetic acid
.
Le A 26 914 - 46 -
. ~ :
.
S~S
NH2~C02H
R,S
The title compound i8 prepared in the manner
described in the literature for the corresponding dithio-
lane compound [M.P. Mertes, A.A. Ramsey, J. Med. Chem.
12, 342 (1969)] by bo~ling the compound from Example 11
in ethanol/KOH.
TLC system III : R~ = 0.05
TLC system V : Rf = O . 35
Exam le 13
N-tert.-butoxycarbonyl-2-R,S-~mino-2-~2-(1,3-dithiano)]-
acetic acid ~
S~S
E~o r ~ NH~C02H
*
R,S
The title compound i~ obtained analogously toExample 5 by xeacting the compound from Example 12 with
di-tert.-butyl carbonate.
TLC system I : Rf = O .1 9
TLC system II : R~ = 0.12
TLC system IV : Rf = O . 55
TLC sy~tem Y : R~ - 0.86
MS-DCIg m~e 234 (M+H); m/e 311 (~+NH~); m/~ 255
~H-MMR ~250 ~Hz, DMSO) s ~: .12.88 (broad~
7.0 ~d, lH); 4.44 (m, 1~);
4.11 ~d~ lH); 2090 (m, 2H3;
Le A 2~ 2~ ~ 47 ~
~ ' ' ': `' '
:
~ :
2.70 ~m, 2H); 1.86 (m, 2H);
1.38 (s, 9H).
Example 14
Ethyl N-acetyl-2-R,S~amin ~ -[2-t1,3-dithiano)]acetate
X~
C~3-C-NH OOC2~5
20 g ~90.5 mmol) of the compound from Example 11
are dissolved in 400 ml of dioxane, and 10 ml (69 mmol)
of triethylamine and finally 7.7 ml (81.3 mmol) of acetic
anhydride are added. The mixture is stirred at room
temperature overnight, dioxane is stripped off and the
residue is triturated with ether~bicarbonate solution.
The residue is isolated, tritura~ed ~ain with diethyl
ether, filtered off with suction and dried.
Yield; 20.9 g (87.8% of theory)
TLC system IV : R~ = 0.76
TLC system V : Rt = 0.89
HPLC system II : ~ = 1.78 min
MS-DCI; m/e 264 (M+H)
Example 15
Ethyl N-acetyl-2-R,S-amino-2-[2-(1,3-dithiolano)~a~etate
I~
O S~S
CH3-C-NH~COOC2H5
~R,S
The title ~ompound i8 obtained analogously to
E~ampl~ 14 8t rting from ethyl 2 R,S-amino 2-12-(1,3-
Le A ?Ç 914 - 48 -
,
r~ ~
dithiolano)]acetate [M.P. Mertes, A.A. Ramsey, J. Med.
Chem. 12, 342 (1969)].
TLC system I : Rf = 0.75
TLC system II : . R~ = 0.67
HPLC ~ystem II : ~ = 1.73 min
MS-DCI : m/e = 250 (M+H)
Exam le 16
Ethyl N-ethoxycarbonyl-L-phenylal~nyl-{2-R,S-amino-2-~2-
(1,3-dithiano)]}acetate
S~S
EtOC-N~ ~ CONH~COOC2H5
R,S
The title compound is o~tained analogously to
Example 1 by propanephosphonic acid anhydride coupling of the
compound from Example 11 (3 g; 13 mmol) and N-ethoxy-
carbonyl-L-phenylalanine.
Yield: S g (87.4% of theory)
TLC system I : Rf = O . 87 both
HPLC system II : ~ = 4.58 min stereoi60mers
(~) FAB-MS : m/e = 441 (M~H); m/e = 463 ~M+Na)
Example 17
N-ethoxycarbonyl L-phenylalanyl-{2-R,S-amino-2-~2-(1,3-
dithiano)]}acetic acid
H~C ~ ~:
EtOC N ON o2H
R,S
Le A 26 914 - 49 -
The ti~le compound is obtain~d analogously ~o
5 ExampleX or Example XI by basic hydrolysis of the
compound from Example 16 (4 g; 9,1 mmol) and standard
work-up,
Yield: 3,7 9 (97% of theory)
TLC system IV : Rf = 0.63 (isomer A)
: Rf = 0.54 ~;somer B)
~) FAB-MS : mle 413 ~M~M); m/e 419 ~M~Li).
Preparation Examples t general formula I J Ia, Ib,
Ic and Id)
ExamDle 18
Methyl-N-tert.-butoxycarbonyl-L-t3,5-diiodo-4-
methoxy)p~enylalanina~
OCH3
~ J
Boc-NH ~ OOCH3
Boc-L-3,5-diiodo-tyrosine tl,O g, 1,87 mmol; Bissendorf
~iochemicals) was stirred wi~h methyl iodide ~352 ~1,
5,61 mmol) and pota~sium carbonat (0.78 9, 5.61 mmol)
in dimethylformamide (10 ml DMF for 3 day~, pro~ected
from moi~rs wi~h a drying tube. After ~vaporotion ~he
reac~ion mixture was d;lut~d with wa~er (150 ml) and
~xtrac~ed with ~ther three ~imes ~50 ml each). The com-
bined organic phase i~ dri~d over codium ~ulphate and
~vaporated.
~5 ..
Le A 2~ 914 - 50
;
3d~
Yield: 1.12 9 () 100 % of ~heory, con~ains DMF)
TLC-Sys~em IV: Rf = 0.89
H-NMR (250 MHz/ DMSO): ~ = 7,7 (s~ 2H); 7,28 (d, lH);
4,15 (m, lH): 3,71 (g, 3H);
3,62 ~s, 3H); 2,95 (dd, lH);
2,74 (dd, lH); 1~33 (s, 9H).
Example 19
N-~er~.-Bu~oxycarbonyl-L-(3,5-diiodo-4-me~hoxy)phenyl-
alanine
~ ~ OCH3
~ J
Boc-NH C02H
The ~ compound is ob~ained analogously ~o Example
X or Example XI by basic hydrolysis of ~he compound from
Example 18 (0.45 9, 0.80 mmol) and s~andard work-up.
Yield: 0.33 9 (75 % of ~heory)
TLC-Sys~em IV: Rf = 0.56
~5
Le A 26 914 - 51 -
- : :
: ~
: .
`` 2 ~ J ~
lH-NMR (250 MHz, DMSO): S = 12,65 ~broad, lH); 7,71
(s, 2H): 7,09 (d, lH), 4,08
~m~ lH); 3,70 ~s, 3H); 2,96
~dd, lH~; 2,69 ~dd, lH);
1~32 ~s, 9H).
~m~a~
N-tert.-butoxycarbonyl-2-R,S-amino-2-[2-(1,3 dithio-
lano)]-acstyl-4S-amino-3S-hydroxy-S~cyclQhexyl-penta-
noyl-L-i~oleucine-(2-picolyl)amide
B o c - NHXC O - N H~C O - N HX~; - NH~3
20 ml of methylene chlo~ide and 18.3 ml
(105 mmol) of diisopropylethylamine are added to ~.4 g
(15.7 mmol) of the compound from Example 5 and 7.3 g
(15 mmol) of the compound from Example 4. The solution i~
25 stirred and cooled to -20C (dry ice~acetone). 11.7 ml of
a 50% strength solution of propanephosphonic acid anhydride in
methylen~ chloride (Hoechst AG) are added dropwise at
this temperatur0 an~ the mix~ure is allow~d to coma to
room temp~rature overnight. The methylone chloride i~
stripped o~f on a rotary evaporator and the r~du~ i6
~6
Le A 26 914 - 52 ~
: . . .
2 ~
taken up in ethyl acetate. The organic phase is extracted
by sha~ing three time3 each with saturated sodium hydro-
gen carbonate solution, buffer solution pH 4 (Merck,
~rticle No. 9435) and saturated sodium chloride solution,
dried over sodium sulphate, concentrated and dried in a
high vacuum.
Yield : 9 g ~79.8% of theory)
(~) FAB-MS ~ mte 680 (M+H); m/e 686 (M~Li)
TLC system III : Rf = 9.59
TLC systam IV 5 Rr = 0.67
HPLC sy~tem IX : Ft = 6.16 min
Example II
2-R,S-amino-2-[2-(1,3-dithiolano)]-acetyl-4S-amino-3S-
hydroxy~5-cyclohexyl-pentanoyl-L-isoleucine-(2-picolyl)-
amide dihydrochloride
sl~ ~3 .
HCl x NH2 X CO-NH ~ O-NH ~ -NH ~ x HC1
150 ml of 4N hydrochloric acid in dioxane are
added with ice cooling to 9 g (13.24 mmol) of the com-
pound from Example I and the mixture i~ ~tirred for ~
hour3. The dioxane/hydrochloric acid mixture i~ removed
by distillation on a rotary evaporator. The product
obtained by repeated coev~poration with diethyl ether i~
filtered off with suction, washed with diethyl ether and
dxied over potas~i~m h~droxide in a de~iccator.
Yield s B.6 g (100% o~ theory)
~) FAB-~S : m/e 580 (~H); m/e 602 t~+Na)
Le A 26 214 53
.
2 ~
TLC system III : Rf = O . 38 (isomer A)
Rf = 0.33 (isomer B)
TLC system IV : Rf = O . 21
ExamE~
N-tert.-butoxycarbonyl-L-phenylalanyl-{2-R,S-amino-2-[2-
(1,3-dithiolano)]}-acetyl-4S-amino-3S-hydroxy-5-cyclo-
hexylpentanoyl-L-isoleucine-(2-picolyl)amide
H ~ X ~
B o c - N C Ct NH O - NH~' C O - NH O - NH~N~
R, S 4
OH
50 ml of methylene chloride and 11 ml (63 mmol~
of diisopropylethylamine are added to S.7 g (9 mmol) of
~he compound from Example II and 3.1 g (11.7 mmol) of N-
tert.-butoxycarbonyl-L-phenylalanine. Tha solution i8
stirred a~d cooled to -20C (dry ice/acetone). 8.2 ml
(12.6 mmol) of a 50% strength solution of propane-
~hosphonic acid a~hydride in methylene chloride /Hoechst AG)
are added dropwise at thi6 temperature and the mixture is
allowed to come to room temp~rature overnight. 100 ml of
methylene chloride are added and the mixture is extracted
by shaking three tlmeR with saturated ~odium hydrogen
carbonate solution, 3 times with buffer solution pH 7
~Merck, Article No. 9439) ~nd once wit~ hal~-saturated
sodium chloride ~olu~ion. The organic phase is dried over
~odium sulphate, con~en~rated and dried in a high v~cuum.
Yiald t 7.2 g (96% of thQo~y)
(+) F~B-~S s m~e 827 (~+H)
TLC ~y~tem I s R~ - O.42 (is~mer A)
Le A 26 914 54
~ , j ,
2 Q ~ 2 ~1 ~ 8
Rf = 0.38 (isomer B)
TLC system III : Rf = 0.60 (isomer A)
Rf = 0.56 (isomer ~)
HPLC system II : ~ = 14.34 min (isomer A)
~ = 12.59 min (isomer ~)
Example _V and Example V
N-tert.-butoxycarbonyl-L-phenylalanyl{2-s-amino-2-[2
(1,3-dithiolano)]}-acetyl-4S-amino-3S-hydroxy-5-cyclo-
hexylpentanoyl~L-isoleucine-(2-picolyl)amide(ExampleIV)
N-tert.-butoxycarbonyl-L-phenylalanyl{2-R-amino-2-[2-
(1,3-dithiolano)]}-acetyl-4S-amino~3S-hydroxy-5-cyclo-
hexylpentanoyl-L-isoleucine-~2-picolyl)amida (Example V)
~S~ ~
Boc-NH ~ 0--NH X 0-NH ~ 0-NH ~ -NH
* S = Example 9 (isomer A)
* R = Example 10 (isomer B)
7.2 g of the ~ompound from Example III are
~eparated on silica gel S0 (Merck, Art. No. 9385) 0.040 -
0.063 mm (230 - 400 me~h) using a step gradient of
methyle~e chloride/methanol ~100/0; 99/1; 98/2; 97/3;
95/5; 9/1). Detection i8 carried out at 214 nm, and the
fraction checking by ~hin layer ~nd high pressure liquid
chromatography. After oombining and removing the ~olvent
on a rotary ~Yapor~tor~ ~he following ~re obtained:
3.1 g of ~he compound ~rom ~xample IV (isomer A ~ S)
O.6 g of the compound from Example IV and ~xample V
Le A ?6 914 55 _
' ' ' .
,
2~
(mixed fraction of A and B)
2.91 g of the compound from Example V ~isomer B . R)
Analytical data for isomer A~(~xample IVl
TLC system III : R~ = a . 60
HPLC sy~tem II : ~ = 14.34 min
~+) FAB-MS : m/e 827 (M+H)
Analytical data for isomer B (Example V)
TLC ~ystem III : R~ = 0.56
HPLC 6y6tem II : ~ = 12.59 ~in
(+) FAB-MS : m/e 827 (M+H)
Example VI
N-tert.-butoxycarbonyl-2-R,S-amino-2~t2-(1,3-dithio-
lano)3-acetyl-~S-amino-3S-hydroxy-5-cyclohexylpentanoyl-
L-leucine ethyl e~ter
r~ ~ I
S~S
Bo c - NHJ~CO- NH~O- NH~COOC 2H5
R,S 4
OH
The title compound i~ obtained analogously to
E~le I by propanephos~honic acid anhydride coupling of 4 g
(14.3 mmol) of the compound from Example 5 and 4.6 g
(13 mmol) of the compound from Example 7. The crude
product ~8.6 9) is fil~ered through 400 g of silica gel
60 (Merck, ~rticls No. 9385; 0.040 - 0.063 mm) u~ing a
stepped gradient of m~thylene chlorideJmethanol (100/0;
98/2; 95/5). The product-containing fractions are ~om-
bined and co~contrat~d.
~ield s 5.7 g (66% of theory)
TLC ~y~tem IV s R~ = 0.85
Le A 26 914 -56-
2 ~1 8 ~3
HPLC system II : ~ = 17.91 min
(+) FA~MS : m/e 618 (M~H); m/e 640 (M+Na)
Example VII
2-R,S-amino-2-[2-(1,3-dithiolano)]-acetyl-4S-amino-3S-
hydroxy-5-cyclohexylpentanoyl-L-leucine ethyl ester
hydrochloride
S~S ~ ~
HCl x NH2 ~ CO-NH ~ o-NH ~ C2H5
OH
The title compound i~ obtained analogously to
Example II by removal of the N-tert.-butoxycarbonyl
protecting group from 5.3 g (8.6 mmol) of the compound
from Example VI.
Yield : 4.7 g (85% of theory)
TLC system III : ~s ~ 0.71 (isomer A)
R~ = 0.64 (isomer B)
(+)F~B-M5 : m/e 518 (M+H); m/e 540 (M+Na)
Example VIII
N-tert.-butoxycarbonyl-L-phenylalanyl-{2-R,5-amino-2-[2-
(1,3-dithiolano)]}-acetyl-4S-amino-3S-hydroxy-5-cyclo-
hexylpentanoyl-L-leucine ethyl ~ster
H ~ H ~ H ~
~c-N O--N O-NH ~ O-N OOC2H5
R,S OH
The tltle compound i8 obtained analogously to
~le III ~y propanephosphonic ~cid anhydride couplings of
Le A_2~ 914 57
1.7 g (6.6 mmol) of N-tert.-butoxycarbonyl-L-phenyl-
ala~ine and 2.8 g (5 mmol) of the compound from Example
VII.
Yield : 3 g (78% of theory)
TLC system I s Rr = 0.65 ~isomer A)
R~ = 0.57 (isomer B)
HPLC system II : ~ = 35.58 min (isomer Al
~ = 31.64 min (isomer B)
(+) FAB-Ms : m/e 765 (M+H); m/e 787 (M+Na)
Example IX
N-ethoxycarbonyl-L-phenylalanyl~{2-R,S-amino-2-t2-(1,3-
dithiolano)]}acetyl 4S-~mino-3S-hydroxy-5-cyclohexyl-
pentanoyl-L-leucine ethyl e~ter
~05X~
E~OC-N O--NH O-NH O-NH COOC2H5
H 4 1
R,S OH
The title compound i3 obtained analogou~ly to
Example III from 1.6 g (6.6 mmol) of N-ethoxycarbonyl-L-
phenylalanine and 2.8 g (5 mmol) of the compound from
Example VII.
Yield : 2.3 ~ (62% of theory~
TLC system I : Rf - O. 50 (both isomers A~B)
HPLC ~ystem II : R~ = 18.66 (i~omer A)
R~ = 15.31 (isomer B)
(+)FAB-MS : mJe 737 (~+N); m/e 759 (M+Na)
Example X
N-tert.-bu~oxycarbonyl-~-phenylalanyl-{2--R,S-amino-2-t2-
(1,3-dithiolano)3}acetyl-4S-amino~3S-hydroxy-5-eyclo-
Le A 2~ 914 -5&-
- ~0184~8
hexylpentanoyl-L-leucine
~SxS ~ ~
Boc-N~C(~NH CO-NH~CO-NH C02H
OH
4.3 ml of an aqueous 1 N sodium hydroxide solu-
tion are added at room temperature to 3 g (3.9 mmol) of
the compound from Example VIII dissolved in 30 ml of
dioxane.The mixture is ~tirred at room temperature for 2
days and then rendered acidic with 1 N aqueous hydro-
chloric acid. The organic solvent is stripped off on
rotary e~aporator, the residue is filtered off and
resuspended in water/acetonitrile 95/5, and the suspen-
sion is frozen and lyophilised.
Yield : 2.9 g (90~ of theory~
(~)FA~-MS : m/e 803 (M+2Na-H); m/e 819 (M~Na+R-H).
Example XI
N-ethoxycarbonyl-L-phenylalanyl-{2-R,S-amino-2-[2-(1,3-
lS dithiolano)]}acetyl-4S amino-3S-hydroxy-5-cyclohexyl-
pentanoyl-L-leucine
1l ,~3~xs ~ ~
E~OC- O--N O-N O-N 02H
OH
The title compound i8 obtained analogou~ly to
Example X by baRic hydroly~is of 2.3 g (3.1 mmol) of the
compound ~rom Ex2mple I~.
Yield s a .5 ~ (87~ of theory)
Le A 26 214
-59-
, :'
- ~ .
,~
(~)F~B-MS : m/e 775 (M+3Na-2H); m/e 791 tM+2Na+X-2H)
Example XII and Example XIII
N-ethoxycarbonyl-L-phenylalanyl-{2-S-amino-2-[2-(1,3-
dithiolano)]}acetyl-4S-amino-3S-hydroxy-5-cyclohexyl-
pentanoyl-L-isoleucine-(2-picolyl)amide (Example XII)
N-ethoxycarbonyl-L-phenylalanyl-{2~R-amino-2-[2-(1,3-
dithiolano)]}acetyl-4S-amino-3S-hydroxy-5-cyclohexyl-
pentanoyl-L-isoleucine-(2-picolyl)amide (Example XIII)
H3C~O-C-N~O 3NH~ON11~0NHX~NH~
*S = Example XII (isomer A)
*R = Example XIII (i~omer B)
The title compounds were obtained by chromato-
graphic separation (analogous to Example IV and Example
V) of the isomer mixture synthesized from N-ethoxy-
carbonyl-L-phenylalanine and the compound from Example II
analogously to Example III. Example XII elutes first
(isomer A, S-isomer), then Example XIII (isomer Bo R-
isomer).
Analytical data for Example XII L~somer A)
~LC syfitem III : Rf - 0.51
HPLC ~ystem II : R~ = 7.35 min
~ AB-NS : m/e = 799 (M~H); m/e = 821 (~Na)
Analytical data for Exam~le XIII ~ifiomer B~
TLC system ITI ~ R~ = 0.47
HPLC sy~tem II : Ft = 5.81 ~in
Le A 26 914 -60-
(+)FAB-MS : m/e = 799 (M+H); m/e = 821 (M+Na)
Example XIV and Example XV
N-tert.-butoxycarbonyl-L-(l-naphthyl)alanyl-{2-S-amino-
2-[2-(1,3-dithiolano)]}acetyl-4S-amino-3S-hydroxy-5-
cyclohexylpentanoyl-L-isoleucine-(2-picolyl)amide
(Example XIV)
N-tert.-butoxycarbonyl-L-(l-naphthyl)alanyl-{2-R-amino~
2-12-(1,3-dithiolano)]}acetyl-4S-amino-3S-hydroxy-5-
cyclohexylpentanoyl-L-isoleucine-(2-picolyl)amide
(Example XV)
Bo c~ oNH~oNHX~;NH~
~S = Example XIV (isomer A)
*R = Example XV (isomer B)
The title compound6 were obtained by chromato-
~raphic separation (analogous to Example IV and Example
V) of the isomer mixture synthesized from N-tert.-butoxy~
carbonyl-L-(l-naphthyl)alanine and the compound from
Ex~mple II analogously to Example III. Example XIV elutes
firs~ (isomer A, S-isomer), then Example XV (isomer B,
R-isomer).
Analy~ic 1 data for Example XIV (isomer A~
TLC system I : R~ = 0.59
HPLC ~ystem II : F~ = 24.34 min
~)FAB-~5 3 m/e = 877 (M+H~; m/e ~ 899 (M~Na~
Analytical data for Example XV (isomer~B~
Le A 26 914 -61-
. ~ :
'
2 ~
TLC system I : R = 0 55
HPLC system II : ~ = 20.49 min
(+)FAB-MS : m/e = 877 (M+H); m/e = 899 (M+Na)
Exam~le XVI and Exam~l~e XVII
N-tert.-butoxycarbonyl-L-(4-methoxy)phenylalanyl-{2-S-
amino-2-[2 (1,3-dithiolano)]}acetyl-4S-amino-3S-hydroxy-
5-cyclohexylpentanoyl-L-isoleucine-(2-picolyl)amide
(Example XVI)
N-tert.-butoxycarbonyl-L-(4-methoxy)phenylalanyl-{2-R-
10amino-2-[2-(1,3-dithiolano)]}acetyl-4S-amino-3S-hydroxy-
5-cyclohexylpentanoyl-L-isoleucine-(2-picolyl)amide
~Example XVII) ICH3
~S~ ~
B o c - NlC O--NH~C ONH~C ONHX~NH~3
~S = Example XVI (i40mer A)
*R = Example XVII (isomer B)
15The title compounds are obtained by chromato-
graphic separation (analogou~ to Example VI and Example
V) of the isomer mixture synthe~ized from N-tert.-butoxy-
carbonyl-L-(4-methoxy)phenylalanine and the compound from
Example II analogously to Example III. Example XVI
(i~omer A, S-i~omer) elutes first, then Example XVII
(isomer B, R-i~omer)0
Analytical data for Example XVI ~i~somer A~
TLC syst~m I s R~ - O.59
HPLC ~ystem II : ~ 3 11 ~ 77 min
Le A 26 914 -62-
2~
(~)FAB-MS : m/e = 857 (M+H); m/e = 879 (M+Na)
Analytical data for Example XVII ¢isomer_B)
TLC system I : Rf = O . 56
HPLC system II : ~ = 10.49 min
(~)FAB-MS : m/e = 857 (M+H); m/e = 879 (M+Na)
Example XVIII
Methyl N-tert.-butoxycarbonyl-2-R,S-~nino-2-[2-(1,3-
dithiolano)]-acetyl-~S-amino-3S-hydroxy-5-cyclohexyl-
pentanoate
1~ ~
s~
1* 1 3
B t~ c - NH~C O NI~ ~ O O C H 3
$F~,S OH
The title compound is obtained pure analogously
to E~le I by propanephosp~lonic aci~ anhydride ooupling of
2.2 g (7.8 mmol) of the compound from Example 5 and 2.4 g
(6.5 mmol) of methyl 4S-amino-3S-hydroxy-5-cyclohexyl-
pentanoate hydrochloride [obtainable by esterification of
4S-amino-3S-hydroxy-5-cyclohexylpentanoic acid (J. Boger
et al., J. Med. Chem. 28, 1779 l1985) or P.F. Schuda et
al., J. Org. Chem. 1988, 53, 873-875) with diazomethane
and precipitation a~ the hydrochloride] and subsequent
silica gel chromatography.
Yield : 3.6 g (88% of theory)
TLC sy~tem IV ~ Rf = 0.88 l i~omer~ cannot
HPLC ~ystem II : ~ = 7.06 min J be differentiated
(+)FAB-MS ~ m/e 497 (M+Li); m/e 441; m/e 397
Exam~le XIX
Methyl 2-R,S-amino-2-[2-~1,3-dithiolano3~-~cetyl-4S-
Le A 26 914 -63-
,
;
--``` 2 ~ 8
amino-3S-hydroxy-5-cyclohexylpentanoate hydrochloride
r~ ~
S~S ~
2 ~R S ~ COOCH~
OH
The title compound i8 obtained analogously to
Example II by removal of the Boc protecting group from
3.5 g (7.1 mmol) of the compound from Example XVIII.
Yield : 3.0 g ~100~ of theory)
TLC ~ystam III : Rl = 0.40 (isomer A)
0.34 (isomer B)
(+)FAB-~S : m~e 391 (M+H); m/e 397 (M+Li~.
Example_XX and Exa~m~le XXI
Methyl N-ethoxycarbonyl-L-phenylalanyl-{2-S-amino-[2-
(1,3-dithiolano)~}-acetyl-4S-amino-3S-hydroxy-5-cyclo-
hexylpentanoate (Example XX)
Methyl N-ethoxycarbonyl-L-phenylalanyl-{2-R-amino-[2-
(1,3-dithiolano)]}-acetyl-4S-amino-35-hydroxy-5-cyclo-
hexylpentanoate (Example X~I)
~r~ ~
O I SyS ~
Et.OC-NH /--CONH l CONH ~ COOCH~3
* 4
OH
~S - Example XX ~i~o~er A)
~R - ~x~mple XXI tisomer B~
The title ~ompound~ ar~ obtained by chromato-
graphic ~eparat~on ~a~alogou~ to Example IV and Example
Le A 26 914 -64-
2~13~
V) of the isomer mixture obtained from 1.3 g (5.5 mmol)
of N-ethoxycarbonyl-L-phenylalanine and 1.8 g (4.6 mmol)
of the compound from Example XIX analogously to Example
III. Example XX (isomer A, S-isomer) elutes first, then
S ~xample XXI (isomer B, R-isomer).
Analytical data for isomer A (Example XX):
TLC sy~tem I : R~ - 0~90
HPLC ~ystem II : ~ = 7.64 min
(+)FAB-MS : m/e 616 (M+Li); m/e 632 (M+Na).
Analytical data for iso~mer B ~Example_XXI~:
TLC system I : R~ = 0.86
HPLC system II : ~ = 6.63 min
(+)FA~-MS : m/e 616 (M+~i); m/e 632 (M+Na).
Exam~le XXII and Example XXIII
Methyl N-tert.-butoxycarbonyl-L-(4-methoxy)phenylalanyl-
{2-S-amino-2-[2-(1,3-dithiolano)]}-acetyl-4S-amino-3S-
hydroxy-5-cyclohexylpentanoate (Example XXII)
Methyl N-tert.-butoxycarbonyl-L-(4-methoxy)phenylalanyl-
{2-R-amino-2-r2-(1,3-dithiolano)]}-acetyl-4S-amino-3S-
hydroxy-5-cyclohexylpentano~te (Example XXII)
~r' ,~3
8cc-N CON~ ~ CONH ~ OOCH3
0~
*S 3 ~xample X~II (isomer A)
*R = Example XXIII (i~omer ~)
~h~ title compounds are obtained analogously to
Le A 26 914 -65-
2 0 :3
Example XX and Example XXI starting from 1.9 g (6.3 mmol)
of N-tert.-butoxycarbonyl-L-(4-methoxy)phenylalanine and
2.4 g (5.7 mmol) of the compound from Example XIX.
Analytical data for isomer A (ExamPle XXII!:
TLC system II : ~f = 0.58
HPLC system II : ~ = 12.33 min
~+)FAB-M5 m/e 668 (M+H)
Analytical data for isomer B ~Example XXIII):
TLC system II : Rs = 0.56
HPLC system II : ~ = 11.25 min
(+)FAB-MS m/e 668 (M+H)
Example XXIV
N-ethoxycarbonyl-L-phenylalanyl-{2-R,S-amino-2-[2-(1,3-
dithiolano)]}acetyl-4S-amino-3S-hydroxy-5-cyclohexyl-
pentanoic acid
~3 r~ ~
Et.OC-NI ~ \ CONH X CONl~ ~ co2H
*R,S 4
OH
460 mg (0.754 mmol) of a mixture of the compounds
from Example XX and Example XXI are dissolved in 20 ml of
dioxane/water 1:1, 180 ~1 (1.4 eq) of 6 N NaOH are added
and the mixture is stirred at room temper~ture overnight.
The mixture is ad~u ted to pH 3 with 1 N HCl and dioxane
is removed on a rotary evaporator. The precipitate is
filtered off with ~uotion, washed with a little water ~nd
dried over KOH in a deAiccator.
Yield : 333 mg (73.5% of theory)
TLC sy~tem I : R~ = O.34
Le A 26 914 -66
~a~ 8~
TLC system IV: R = 0 74
(+)FAB-MS : m~e 634 ~M+K); m/e 672 (M+2K-H).
Example XXV
N-tert.-butoxycarbonyl-L-(4-methoxy)phenylalanyl-{2-R,S-
amino-2-[2-(1,3-dithiolano)]}acetyl-45-amino-3S-hydroxy-
5-cycloh0xylpentanoic acid
B o c - NI~C ONH~C ONE~CO 2 H
OH
The titla compound is obtained analogously to
Example XXIV from 790 mg (1.18 mmol) of a mixture of th~
compounds from Example XXII and Example XXIII.
Yield : 560 mg (73~ of theory)
TLC system I : R~ = 0.18
TLC sy~tem III : Rf = 0.05
TLC system IV : Rf = 0 . 82
(~)FAB-MS : m/e = 660 (~+Li); m~e = 676 (M+Na).
Example XXVI
N-tert~-butoxycarbonyl-2-R,S-amino-2-[2-(1,3-dithio-
lano)]-acetyl-4S-amino-3S-hydroxy-5-cyclohexyl-pentanoyl-
(2S-methyl)butylamide
~ ~ H C
S~S ~ '3Hr
Boc-N ONH~A~^~ON
~R,S q ¦
0~
The titlo compound is obtained ~nalogously to
E~le I by propanephosphonic acid anhydride coupling of -
Le A 26 914 -67-
.
,
. ,
.
--` 2 ~
3.8 g (12 mmol) of the compound ~rom Example 9 and 4 g
(14.4 mmol) of the compound from Example 5.
~ield : 5.5 g (82.5~ of theory)
TLC system I : R~ = 0.52 ~ isomers cannot
HPLC system II : Rt = 13.38 min J be differentiated
(+)FAB-MS : m/e 545 (M~H); m/e 552 (M+Li);
m/e 564 (M~N~)
Example_XXVII
2-R,S-amino-2-(2-(1,3-dithiolano)]-acetyl-4S-amino-3S-
hydroxy-5-cyclohexylpentanoyl-(2S-methyl)-butylamide
hydrochloride
s~--s ~ H3
1 3 12
}IC 1 x N112f~CONH CONH~
~R,S 4 1
OH
The title compound i~ obtained analogously to
Example II by removing the Boc protecting group from
5.4 g (9.9 mmol) of the compound from Example XXVI.
Yield : 4.2 q (95~ of theory)
TLC sy~tem III : Rs = 0.41 (isomer A)
Rf = 0.35 (isomer 3)
Example XXVIII and Exam~le XXIg
N-ethoxy-carbonyl-L-phenylalanyl-~2-S-amino-2-[2-~1,3-
dithiolano)]}acetyl-4S-amino-3S-hydroxy-5 cyclohexyl-
pentanoyl-(2S-methyl)-butylamide (Ex~mple XXVIII)
N-ethoxy-carbonyl-L-phenylalanyl-{2-R-amino-2-[2-(1,3-
dithiolano)]}acetyl-4S-amino-3S-hydroxy-5-cyclohexyl-
pentanoyl-~2S-methyl)-~utyl~mide (Example XXIX)
Le A ?6 914 -68-
.
H3C ~ O-C-N ~O- NH ~ CONH ~ CONH
H 4
OH
~S ~ Example (XXVIII) (isomer A)
~R = Example (XXIX) (isomer B)
The title compounds were obtained by chromato-
graphic separation (analogous to Example IV and Example
V) of the isomer mixture obtained from 2.1 g (4.7 mmol)
of the compound from Example XXVII and 1.3 g (5.5 mmol)
of N-ethoxycarbonyl-L~phenylalanine analogously to
Example III. Example XXVIII (isomer A, S-isomer) elutes
first, then Example XXIX (isomer B, R-isomer). A mixed
fraction of Example XXVIII and Example XXIX is
additionally obtained.
Analvtical data__for isomer ~lExample XXVIII~:
TLC system I : Rf = O . 6 8
HPLC system II : ~ = 13.27 min
(+)FAB-MS : m/e = 671 (M-~hi); m/e = 687 (M+Na).
Analytical data for isomer ~ IExample XXIX):
TLC system I s R~ = 0.65
HPLC system II : ~ = 11.61 min
(+)FAB-MS s m/e = 671 (M+~i); m/e = 687 (~Na).
Example XXX and Example XXXI
N-tert. butoxycarbonyl-L-(~-methoxy)phenylalanyl-{2-S-
amino-2-[2-(1,3-dithiolano)]acetyl-45-amino-3S-hydroxy-
5-cyclohexylpentanoyl-(2S-methyl)butylamid~(Example~XX)
N-tert.-butoxycarbonyl-L-(4-methoxy)ph~nylalanyl-~2-R-
Le_A 26 9..14 -69-
,
amino-2-[2-tl,3-dithiolano)]acetyl-4S-amino-3S-hydroxy-
5-cyclohexylpentanoyl-(2S-methyl)butylamide (Example
XXXI)
~ 3 ~
Boc-M CONH CONH~CONH
OH
~S ~ Example XXX (isomer A)
*R a Example XXXI (l~omer B)
The title compoundq are obtained analogously to
Example XXVIII and Example XXIX starting from the com-
pound from Example XXVII and N-tert.-butoxycarbonyl-L-(4-
methoxy)phenylalanine. A large mixed fraction also
occurs.
Analytical data for Example XXX:
T~C system II : R~ = 0.38
HPLC system II : ~ = 22,80 min
(+)FA~-MS m/e = 729 (M+~i)
Analy~ical data for Example XXXI:
TLC system II : Rf = 0 . 34
HPLC system II : Rt - 22,80 min
(+)FA3~NS m/e = 729 (M+Li)
Example XXXII and Example XXX~II
N-ethoxycarbonyl-L-(4-methoxy)phenylalanyl-~2-5-amino-2
[2-~1,3-dithiolano)]}acetyl-4S-amino-3S-hydroxy-5-~yclo-
hexylpent~noyl-L-i~oleucine-(2-picolyllamide (~ample
XXXII)
N-etho~ycarbonyl-L-(4-methoxy)phenylalanyl ~2-R-amino-2-
Le A 26 914 _70_
~ 3 ~ 8
[2-~1,3-dithiolano)~}acetyl-4S-amino-3S-hydroxy-5-cyclo-
he~ylpentanoyl-L-isoleucine-(2-picolyl)amide (Example
XXXIII)
E~OC-N ~ CONH X CONII ~ ONH ~ NH ~
~S -- Example XXXII (isomer A)
~R = Example XXXIII (isomer B)
The title compounds are obtained by chromato-
graphic separation (analo~ous to Example IV and Example
V) of the isomer mixture synthesized from N-ethoxy-
carbonyl-L-(4-methoxy)-phenylalanine and the compound
from Example II analogously to Example III. Example XXXII
(isomer A, S-isomer) elutes first, then Example XXXIII
(isomer B, R-isomer). -
Analytical data ~ g~er A (Ex_m~le XXXTT..):
TLC system III : R~ = 0.72
TLC system IV : R = 0~79
HPLC ~y~tem II : ~ = 7.62 min
(+)FAB-MS : m/e = 829 (M+H); m/e = 851 (M+Na).
Analy~ical data for isomer B (Example XXXIII):
TLC system III : R = 0.69
TLC system IV s Rs = 0.73
HPLC system II s Rt = 6.10 min
(+)FAB-M5 m~e = 829 (M+H); m/e = 851 (M+Na).
Le A 26 914 -71-
- : :
~ .
2~1~4~
Example XXXIV
L-phenylalanyl ~2-R,S-amino-2-[2-(1,3-dithiolano)]}-
acetyl-4S-amino-3S-hydroxy-5-cyclohexylpen~anoyl-L-
isoleucine-(2-picolyl)amide dihydrochloride
~r~ ~
HCl x NH2 ~ CONH X ONH ~ CONH ~ NH ~ HCl
The title compound is obtained analogously to
Example II by removal of the Boc protecting group from
the compound of Example III or a mixture of the compounds
from Example IV and Example V.
Analytical data:
TLC system IV : R~ = 0.43 (isomer A, S-isomer)
Rf = 0 . 40 (isomer B, R-isomer)
Example XXXV
N-tert.-butoxycarbonyl-~-seryl~L-phenylalanyl-{2-R,S~
amino-2[2-(1,3-dithiolano)]}-acetyl-4S-amino-3S-hydroxy-
5-cyclohexylpentanoyl-L-isoleucine (2-picolyl)amide
~ H ~ s
Boc-NH ON CONH ONH ONH ONH ¢~
0}~
The title compound i~ obtained analogously to
Example III ~rom N-tert.-butoxycarbonyl L-~erine and the
compound from ~xample XXXIY.
Le A 25 914 -72-
2 ~ ,L 8 4 ~ 8
Analy~sgL_data:
TLC system II : Rf = 0.16 (isomer A)
R~ = 0.14 (isomer B)
HPLC system II : ~ = 5.04 (isomer B)
~ = 5.70 (isomer A)
(~)FAB-MS m/e 914 (M+H); m~e 936 ~M+Na).
Example XXXVI
N-~3-~3-pyridyl)propionyl]-L-phenylalanyl-{2-R-amino-2-
t2-~1,3-dithiolano)~-acetyl-4S-amino-3S-hydroxy-5-cyclo-
hexylpentanoyl-L~isoleucine-(2-picolyl)amide
~3 1~ ` 1~1
N~NH'J\CONHXONH~ONHXC~ONH ¢~
The title compound i8 obtained analogously to
13xample ~XV by propane~hosphonic acid anhydride coupling of
3~(3-pyridyl)propionic acid with the dihydrochloride
producible from the R-compound of Example V by removal of
the Boc protecting group analogously to Example XXXIV.
Analytical data:
TLC system III : R = 0 47
HPLC ~ystem II : ~ = 6.36 min
(+)FAB-Ms : m/e = 8.60 (M+H); m/e = 882 (M~Na).
Example XXXVII
N-tert.-butoxy~arbonyl-~-phenylalanyl-{2-R,S-amino-2-t2-
t1,3-dithiolano)]~-acetyl-4S-amino-3S-hydr~xy-5-cyclo-
hexylpentanoyl-~ leucine methyl e~ter
Le A 26 ~14 _73_
,
BOC-NR' CONHX~ONH1f ~CONH~;OCH3
RJS 4
OH
The title compound i8 obtained analogously to
Ex~mple VIII, the L-leucine methyl ester being used
instead of the ethyl ester for the synthesis of the
correspondin~ compounds of Examples 6, 7, VI and VII.
Analytical data:
TLC ystem I : Rf = O . 85 (isomer A, S-isomer)
R~ = 0.78 (isomer B, R-isomer)
(+)FAB-MS : m/e 751 (M~H) m/e 773 (~Na);
m/e 651.
Example XXXVIII
N-ethoxycarbonyl-L-phenylalanyl-{2-R,S-amino-2-[2-(1,3-
dithiolano)]}acetyl-4S-amino-3S-hydroxy-5-cyclohexyl-
pentanoyl-L-leucine methyl e~ter
oc-Nl~\coN~oNH~coNH~ocH,
R,S 4
OH
The title compound is obtained analogously to
Example XXXI, ths L-leucine methyl ester being used
instead of the ethyl e~ter f or the ~ynth2sis o~ the
corresponding ctarting compounds of Example~ 6, 7, VI and
VII.
Analytical data:
TLC ~ystem I ~ R~ = 0.81 (isomsr A, S-isomer)
Le A ?6 9I4 -74-
20~g~8
Rf = 0.73 (isomer B, R-isomer)
HPLC system II : ~ = 11.66 min ~isomer B)
~ = 14.08 min (isomer A)
(+)FAB-Ms m/e 723 (M+H); m/e 745 (M+Na).
Example_XXXIX and Example XL
N-tert~-butoxycarbonyl-L-phenylalanyl-{2-s-amino-2-[2-
(1,3-dithiolano)]}acetyl-4S-amino-3S-hydroxy-5-cyclo-
hexylpentanoyl-L-leucine-(3-aminomethyl)benzylamide
(Example XXXIX)
N-tert. butoxycarbonyl-L-phenylalanyl-{2-R-amino-2-12-
(1,3-dithiolano)]}acetyl-4S-amino-3S-hydroxy-5-cyclo-
hexylpentanoyl-L-leucine-(3-aminomethyl~benzylamide
(Example XL)
Boc-N~C30NHX ONH~ONH~ONH~NH2
*S = E~ample XXXIX (isomer ~)
*R = Example XL (isomer B)
O.95 g (1.29 mmol~ of the compound from Example
X and 2.50 ml (19.35 mmol) of 1,3-bi~aminomethylbenzene
are di~solved in 30 ml of CH2C12. The mixture is cooled to
- -2UC and 1.1 ml of a 50% strength solution of n-propyl-
phosp,~ ic acid anhydride in CH2Cl2 (Ho~st AG) are added. The ~:
mixture i8 ~tirred at room temperature overnight, 300 ml
of diethyl ether ~re added and th~ precipitate i~
filtered off with ~uction. ~he residuQ i~ ~tirred with
Le A 26 914 -75-
- :
``" 2 ~
300 ml of diethyl ether again, and the precipitate is
filtered off with suction and dried. The crude product
(1.86 g) is ~eparated on 300 g of silica gel 60 (Merck,
Art. No. 9385) 0.040-0.063 mm (230-400 me-h) using a
stepped gradient of methylene chloride/methanol/aqueous
NH3 solution (25%) 100/0/0; 99/1/0.2; 98/2/0.2; 97/3/0.2
and 95/5/0.2 (0.5 1 each)~
Example XXXIX (isomer A, S-isomer) elutes first,
then Example XL (isomer B, R-isomer).
~a~y~ical data for isomer A_(Example XXXIX~:
TLC system III : R~ = 0.32
(+)FA~-MS m/e = 955 (M+H); m/e = 755
HPLC system II : no elution or a ~ery broad peak.
Analytical data for isomer B (Example XL):
TLC system III : Rf = O . 29
(+)FAB-MS : m/e = 855 (M+H); m/e - 755
HPLC system II no elution or a very broad peak.
Exa~m~le XLI and Example XLII
N-ethoxycarbonyl-L-phenylalanyl-{2~S-amino-2-[2-(1,3-
dithiolano)~}acetyl-4S-amino-3S-hydroxy-5-cyclohexyl-
pentanoyl-L-leucine-(3-aminomethyl)-benzylamide (Example
XLI)
N-ethoxycarbonyl-L-phenylalanyl-{2-R-amino-2-12-(1,3-
dithiolano)]}acetyl-4S-amino-3S-hydroxy-5-cyclohexyl-
pentanoyl-L-leucine-(3-aminomethyl)-benzylamide (Example
XLII)
Le A 26 914 -76-
2~458
o ~coNSXSoNH"~CONH~
E~OC-H OH ~ NH2
~S = Example XLI (isomer A)
~R - Example XLII (isomer B)
The title compounds are obtained analogously to
Example XXXIX and Example XL starting from the compound
of Example XI.
Ana~ytical,data for iso,mer A (ExamPle XLI):
TLC system III : R~ = 0.26
HPLC system II : ~ = 126 min
(+)FA~-MS m/e - 827 (M+H); 849 (M~Na).
Analytical d~ata for i~omer B (Example,XLII~:
TLC system III : Rf = O . 23 :
HPLC sy6tem II : ~ - 106 min
(+)FAB-MS m/e = 827 (M+H); 849 (M+Na)
Example XLIII and Example XLIV
N-ethoxycarbonyl-L-phenylalanyl-{2-S-amino~2-[ 2 - ( 1, 3 -
dithiolano)]}acetyl-~S-amino-35hydroxy-5~cyclohexyl-
pentanoyl-L-isoleucinol (Example XLIII) :,
N-ethoxycarbonyl-L-phenylalanyl-{2-R-amino-2-[2-(1,3-
dithiolano)]~acetyl-4S-amino-3S-hydroxy-S-cyclohexyl-
pentanoyl L-isoleucinol (Example XLIV)
,~
Le A 2~ 77_
:.
.
~ 3~
0 ~3s~s ~ ~
E~OC-N/\CONH~CONH ~CONH~OH
H * 4 ¦
OH
~S - isomer A (Example XLIII)
*R = isomer B ( Example XLIV)
The title compound-q are obtained analogously to
E~le 3 or E~le I by n-propanephosphonic acid anhydride
coupling of the compound from Example XXIV with L-iso-
leucinol hydrochloride as the amine component. The
preparation of the pure stereoisomers is carried out by
silica gel chromatography (analogously to Example IV and
Example V) or, alternatively, also by reversed phase HPLC
on C~ phases (column, for example, Merck Hibar, Cat. No.
51441, LiChrosorb, RP-8 ~7 ~m), size 250-25) using
acetonitrile/water mixtures. Product-containing fractions
from the silica gel chromatography are combined and
concentrated on a rotary evaporator, or with reversed
phase chromatography, scetonitrile i3 removed on a rotary
evaporator and the product is lyophilized.
Analytical data for isomer A (S-isomer)~ Example _LIII) J
~LC system I : R~ = 0.54
TLC system II : Rr = 0 44
TLC system IV : Rf = O . 64
HPLC system II : ~ = 7.10 min
(+)FAB-MS 2 m/e = 695 (~+H); m/e ~ 717 t~a).
Analy~ical data for_isomer B lR-isomer ~xample XLIV!:
TLC sy~tem I : ~ = a .54
TLC system II : R~ ~ 0.42
~ a~3~ 78-
,
~ .
,. ~ , , .". . . . .
.
2 a ~ 8
TLC system IV : Rf = O . 64
HPLC system II : ~ = 5.71 min
(+)FAB-MS : m~e = 695 (M+H); m/e = 717 (M+Na).
Example XLV and Example XLVI
N-ethoxycarbonyl-L-phenylalanyl-{2-S-amino-2-[2-(1,3-
dithiolano)]}acetyl-4S-amino-3S-hydroxy-S-cyclohexyl-
pentanoyl-L-isoleucine i~opropyl ester (Example XLV)
N-ethoxycarbonyl-L-phenylalanyl {2-R-amino-2-[2-(1,3-
dithiolano)]}acetyl~4S-amino-3S-hydroxy-S-cyclohexyl-
pentanoyl-L-isoleucine isopropyl ester (Example XLVI)
E~OC -I~\CONH~CONH~:ONH~O--<
OH O
*S = isomer A (Example XLV)
*R = isomer B (Example XLVI)
The title compounds are prepared analogously to
E~le 3 or I by n-propanephosphonic acid anhydride couplmg
of the compound from Example XXIV using L-isoleucine
isopropyl ester hydrochloride as the amine component. The
preparation of the pure ~tereoi30mers is carried out by
chromatography (as de cribed in Example XLIII and XLIV).
Analytical data for isomer A (S-isomer, Example XLV):
TLC ~ystem I : F~ = O.68
TLC system II : R~ = 0.57
HPLC ~ystem II : Rt = 27.61 min
(~F~B-NS s m/e ~ 751 (M~H); m/e = 773 (~+Na
Le A 26 914 -79-
2 ~
Analytical data for isomer B (R-isomer, Example XLVI ):
TLC system I : R~ = 0.68
TLC system II : Rf = 0.57
HPLC system II : ~ = 22.51 min
S (+)FAB-MS : m/e = 751 (M+H); m/e = 773 (M+Na)
Example XLVII and Example XLVIII
N-ethoxycarbonyl-L-phenylalanyl-{2-S-amino-2-[2-tl,3-
dithiolano)]}acetyl-4S-amino-3S-hydroxy-5-cyclohexyl-
pentanoyl-L-isoleucine-h-valine-(2-picolyl)amide ~Example
XLVII)
N-ethoxycarbonyl-L-phenylalanyl-{2-R-amino-2-t2-(1,3-
dithiolano)]}acetyl-4S-amino-3S-hydroxy-5-cyclohexyl-
pentanoyl-L-isoleucine-L-~aline-~2-picolyl)amide (Example
XLVIII)
E~OC- ~ CONH X CONH ~ CONH ~ NH X ONH
OH
*S = isomer A (Example XLVII)
*R = isomer B (Example XLVIII)
The title compounds are prepared analogously to
~xample 3 or I by n-pro~anephosphonic acid anhydride coupling
of the compound from Example XXIV using L-isoleucyl-L-
valine-(2~picolyl)amide dihydrochloride as the amine
component. The preparation of the pure stereoisomers i8
carried out by chromatography (as de~cribed in Example
XLIII and ~LIV)
Analy~ical data for i~omer A ~S-isomer, Example XLVII~:
TLC ~ystem I s R~ = 0.52
Le A 26 914 80-
-2;~
TLC system II : Rf = 0.40
HPLC system II : ~ = 9.48 min
(+)FAB-MS : m/e = 898 ~M+H)
Analytical data for isomer A ~R-isomer, Example XLVIII):
TLC system I : R~ = 0.52
TLC system II : Rf = 0.40
HPLC system II : ~ = 8.05 min
(+)FAB-MS : mte = 898 (M+H)
Example XLIX and Example L
N-ethoxycarbonyl-L~phenylalanyl-{2-S-amino-2-[2-(1,3- .
dithiolano)]}ace~yl-4S-amino-3S~hydroxy-5-cyclohexyl- :-
pentanoyl ~ -methyl)propylamide (Example XLIX)
N-ethoxycarbonyl-L-phenylalanyl-~2-R-amino-2-t2-(1,3-
dithiolano)]~acetyl 4S-amino-3S-hydroxy-5-cyclohexyl-
pentanoyl -(2-methyl)propylamide (Example L)
El.OC -~CO3NHX~ONH~CONH~
*S = Isomer A (Example XLI~)
~R - Isomer B (Exzmple L)
The t~tle compounds are prepared analogously to
E~le 3 or I by n-propanephosphonic acid anhydride coupling
of the compound from Example XXIV using (2-methyl)-
propylamine as the amine component. The preparation of
the pure ~tereoi~omers i8 carried out by chromatography
(as described in ~xamplas X~III and ~IV).
Le A 26 914 -81-
2~18~8
Analytical data for_isomer A (s-isomer~ Exam~le XLIX):
TLC system I : Rf = 0.66
HPLC system II : ~ = 8.84 min
(+)FAB-NS : m/e = 657 (M+Li)
Analy~ical data for isomer B (R-isomer~ ~xample L):
TLC system I : R~ = 0.66
HPLC system II : ~ = 7.74 min
(~)FAB-~S : m/e = 657 ~M+Li~
Example LI
N-ethoxycarbonyl-L-phenylalanyl-{2-R,S-amino-2-[2-(1,3-
dithiano)J}acetyl-4S-amino-3S-hydroxy-5-cyclohexyl-
pentanoyl-L-isoleucine-(2-picolyl)amide
O ~S~S
EtOC-N ~ CONH'-`CONH ~ ONH'A`~ONH
*R,S OH
The title compou~d is prepared analogously to
E~le 3 or I by n-propanephosphonic acid anhydride coupling
1~ of the compound from Example 17 with the compound from
Example 3 as the amine component.
Analytical data:
TLC system II : Rr = 0.32 ~ the two i~omers
HPLC system II : F~ = 8.34 min ~ cannot be
differentiated
(+)FA~ ~S : m/e - 813 (M~H~; m/e 835 (M+Na~
Example LII and Example~LI~I
N-tert.-butoxycarbonyl-L-(4-m*thoxy)phenylalanyl-[2-S-
amino-2-[2-(1,3-dithiolano)]}acetyl-4S-amino-3S-hydroxy-
Le A 26 914 -82-
2 ~
5-cyclohexylpentanoyl-(m~rpholine)amide ~Example LII)
N-tert. butoxycarbonyl-L-(4-methoxy)phenylalanyl-[2~R-
amino-2-[2-(1,3-dithiolano)]}acetyl-4S-amino-3S-hydroxy-
5-cyclohexylpentanoyl-(~ rpholine)amide (ExalT~le LIII)
~' 1~1
S~s ,~
Boc-NH~ ~CONH~CONH~ON O
OH
*S = isomer A (Example LII)
*R = isomer B (Example LIII)
The title compounds are prepared analogously to
Example 3 or I by n-propanephosphonic acid anhydride coupl m g
of the compound from Example XXV using moxpholine as the
amine component. The preparation of the pure stereo-
isomer~ is carried out by chromatography (as described in
Examples XLIII and XLIV).
Analytical data for isomer A ~S-isomer Example LII!:
TLC system II : Rf = 0 . 39
HPLC ~ystem II : ~ = 7.78 min
(+)FAB-MS : m/e = 7~3 (M+H); m/e = 729 (M+Li);
mie = 745 ~M+Na)
Analytical data for isomer B (R-isomer, Example LIII~-
~LC system II : Rt = O.36
HPLC ~y~tem II : R~ = 7.07 min
(+)FAB-NS : m/e = 723 (~+H); m/e ~ 729 (~+Li);
Le A 26 914 -83-
,
' ' :
2 ~ 8
m/e = 745 (M+Na)
Example LIV and ExamE~ V
N-tert.-butoxycarbonyl-L-(4-methoxy)phenylalanyl-{2-S-
amino-2-[2-(1,3-dithiolano)]}acetyl-4S-amino-3S-hydroxy-
5-cyclohexylpen~anoyl-(4-methyl)butylamide (~xample LIV)
N-tert.-butoxycarbonyl-L (4-methoxy)phenylalanyl-{2-R-
amino-2-[2-(1,3-dithiolano)]}a~e~yl-4S-amino-3S-hydroxy-
5-~yclohexylpentanoyl-(4-methyl)butylamide (Example LV)
~ OC~3
~ ~r~ f~
B o c - Nl~\C ONH~CONH~:ONH~
OH
*S = isomer A ~Example LIV)
*R = isomer B (~xample LV)
The title compounds are prepared analogously to
~xample 3 or I by n-propanephosphonic acid anhydride coupling
of the compound from Example XXV using 3-methylbutylamine
as the amine component. The preparation of the pure
stereoisomers is carried out by chromatography (as
described in Example XLIII and XLTV).
Analytical data for isomer A (S-isom~r, Example LIV):
TLC system II . R~ = 0.38
HPLC system II : Rt = 21~14 min
(+)F~B-MS : m~e = 729 ~N~Li)
Analytical data for isomer B ~Risomer, ExamRle LV)s
TLC system II : R~ = O.35
HPLC system II s R~ = 21.14 min
(~)FAB NS s m/e - 729 ~M+Li)
Le A 26 914 -84-
, ; .
2 ~ 3 8
Example LVI and Example _LVI I
N-tert.-hutoxycarbonyl-L-(4-methoxy)phenylalanyl-{2-S-
amino-2-[2-(1,3-dithiolano)]~acetyl-4S-amino-35-hydroxy-
5-cyclohexylpentanoyl-n-hexyl ~ de (Example LVI)
N-tert.-butoxycarbonyl-L-(4-methoxy)phenylalanyl-{2-R-
amino-2-[2-(1,3-dithiolano)~}acetyl-4S-amino-3S-hydroxy-
5-cyclohexylpen ~ oyl-n-hexylamide (E~le LVII)--
OCH~
S~S ~~
Bo c - Nll/\CONHJ~CON}I~CONH
OH
*S = isomer A (Example LVI)
*R = isomer B (Example LVII)
The ~itle compounds are prepared analogously to
Example 3 or I by n-propanephosphonic acid anhydride oouplmg
of the compound from Example XXV using n-hexylamine
hydrochloride as the amine component. The preparation of
the pure stereoisomers is carried out by chromatography
~as described in Example XLIII and XLIV).
Analytical data for isomer A ~S-isomer, Example_LVI):
TLC system II : Rr = 0 45
HPLC system II : ~ = 34.11 min
(~)FAB-~S : m~e = 737 ~M+H), m/e = 743 (M~Li);
m/e = 759 (~+~a)
TLC 8ystem II ~ Rr = O . 41
HPLC system II ~ R~ = 34.11 min
(+)FAB-Ms s m/e = 737 (M+H3; m~e = 743 tM+Li);
Le A ?6~ -85-
mle = 759 (N+N)
Example LVIII and ExamPle LIX
N-~ert.-buLoxycarbonyl-L-(4-me~hoxy)phenylalanyl-
(2-S-amino-2-[2-(1,3-di~hiolano)]~acetyl-4S-amino-3S-
hydroxy-5-cyclohexylpentanoyl-(N-benzylpiperazine)amide
(Example LVIII)
N-ter,-butoxycarbonyl-L-(4-me~hoxy)phenylalanyl-~2-
R-amino-2-[-tl,3-di~hiolano)~ace~yl-4S-amino-3S-
hydroxy-5-cyclohexylpen~anoyl-(N-benzylpiperazine)amide
(Example LIX
Boc-N ~ CONH X CONH ~ CON N
* 4 ~__~
OH
*S = isomer A (Example LVIII)
*R = isomer B (Example LIX)
The ~itle compounds are prepared analogously ~o
Example 3 or I by n-propanephosphonic acid anhydride
coupling of the compound from Example XXV using N-
benzylpipera7ine as the amine component. The preparationof the pure stereoisomers is carried out by chromato-
graphy (as described in Example XLIII and XLIV).
Analvti al Data for isomer A SS-isomer, Exam~le LVIII~:
TLC-system II: Rf = 0.44
HPLC-6ystem II: R~ = ~1.46 min
(~)FAB-MS: m/e = 818 (M~Li)
Analvtical Da~a for isomer_B ~R-isomerl E~ample LIX):
TLC-~y~tem II : Rf ~ 0144
HPLC system II: R~ = 29.97 min
(t) FAB-MS : m/e = 818 (M~Li)
Le A 26 914 - 86 -
:.
:~ ,
` 23189-7102 ~ ~1 3 ~ ~ 8
Examp1e LX
s
Me~hyl-2-R-amino-2-~2-(1,3-di~hiolano)]-ace~yl-4S-amino-
3S-hydroxy-5-cyclohexylpen~anoa~e
'
S~S
NH2 CONH ~ COOCH3
*R 4 i
OH
The ~itle compound is ob~ained by chroma~ographic
separa~ion of ~he i~omer mix~ure (free base) from
Example XIX on 6ilica gel using a ~epped ~radien~ of
me~hylene chloride~me~hanol/aqueous NH3 solu~ion (Z5X.)
as described in Example XXXIX and XL. The S-isomer
elu~es firs~, then Example LX (R-isomer) .
Analy~cal da~:
TLC-cy~em IlI: Rf = 0,34
(~)FAB-MS: m/e = 391 ~M~H)) m/e = 397 (M~Li).
ExamPle LXI
Me~hyl N-~er~.-bu~oxy~arbonyl-2-R-amino-2-t2-(1,3-
di~hiolano)]-acetyl-4S-amino-3S-hydroxy-5~cyclohexyl-
30 pen~anoa~e
r~ ~
S~S
~ 1 3
Boc NH~ ~ONH ~ COOCH3
*R 4 1
OH
Le A 26 914 - 87-
.,
2 0 ~
The ~i~le compound is obtained analogously ~o
Example 5 by reacting the compound from Example LX with
di-tert.-bu~yl carbonate using triethylamine as base and
dioxane as solvent (analogously to Example 14). The
reaction mixture is diluted wi~h water and lN
hydrochloric acid and extracted with methylene chloride.
The organic phase is separated, dried over sodium
sulphate and concen~rated. The crude product Lhus
ob~ained is purified by flash chromaography (silica/
methylene chloride).
Analvtical data:
TLC-system IV: Rf = 0.88
HPLC-System II: R~ = 7.06 min
(~)FAB-MS: m/e = 497 (M~Li)
Example LXII
N-tert.-butoxycarbonyl-2-R-amino-2-[2-(1,3-dithiolano)~-
acetyl-45-amino-3S-hydroxy~5-cyclohexyl-pentanoic acid
r~ ~
S ~ S
Boc-NH~CONH ~ COOH
*R 4 ¦
0}~
The title compound is obtained analogously to
Example X or Example XI by basic hydrolysis of the
compound from Examplo LXI (2.1 9; 4.28 mmol) and
standard work-up.
Yield: 1.95 3 ~95.6 % of theory)
~5 TLC-System IV: Rf = 0.57
(~)FAB-MS: m/e = 477 (M+H), m/e 421, m/e 377.
Le A 26 91~ - 88 -
,
'
,
lg~3
Example LXIII
N-tert,-butoxycarbonyl-2-R-amino-2-~2-(1,3-
dithiolano)]-acetyl-4S-amino-3S-hydroxy-5-cyclohexyl-
pentanoate
~ ~ J~
~ * ~3 r
Boc-NH ~ ONH ~ ONH ~ OH
* 4 1
R OH
The title compound is obtained analogously to
Example 3 or Example 9 by n-propanephosphonic acid
anhydride coupling of the compound from Example LXII
with L-leucinol hydrochloride as the amine component,
The crude product is purified by chromatography ~ana-
logously to Example IV and Example V).
Analvtical dat~:
TLC-system III: Rf = 0.65
HPLC-System II: R~ = 6.58 min
(~)FAB-MS: m/e = 582 ~M~Li).
Example LXIV
N-tert.-butoxycarbonyl-L-(4-me~hoxy?phenylalanyl-
~2-R-amino-2-t2-rl,3-dithiolano)~acetyl-4S-amino-~S-
hydroxy-5-cyclollsxylpentar~oyl-L-leucinol
Le A 26 914 - 89
~8~$
OCH
1 3
~xS
Boc-NH CONH CONH ~ CONH OH
* 4 1
OH
The title compound is prepared starting with the
compound from Example LXIII by a) removal of the Loc-
protec~ing group (analogously to Example 4) and b)
reacting the hydrochloride thus obtained with Boc-L-(4-
methoxy3phenylalanine ~analogously to Example XVI andXVII, The crude produc~ is purified by chromatography.
Analytical data:
TLC-system IV: Rf = 0,59
HPLC-system Il: Rt = 12.85 min
(~)FAB-MS: m/e = 759 (M~Li).
Example LXV
2-R-amino-2-(1,3~dithiolano)]-acetyl-4S-amino-35-
hydroxy-5-cyclohexyl-pentanoyl-L-isoleucine-(2-picolyl)
amide
~0
r~ r~
NH2 R ONH ~ ONH~CONH
Le A 26 914 - 90 _
. : j; : . .
' ' - ' . :
'
201~8
The title compound is obtained analogously to
Example LX by chromatography starting with the R,S-
mixture of the compound from Example II, The S-isomer
elutes first, then the R-isomer tEx~mple LXV).
Analytical da~a:
TLC-~ystem III: Rf = 0.33
(~)FAB-MS: m/e = S80 (M~H~, m/e 60Z (M+Na).
ExamDle LXVI
L-(~-methoxy)phenylalanyl-~2-R-amino-2-t2-(lJ3-
dithiolano)]Jacetyl ~S-amino-3S-hydroxy-S-cy~lohexyl-
pentanoyl-L-isoleucine-(2-picolyl)amide-dihydrochloride
OCH3
~ H X ~ ~ * HCl
HCl * ~H2 ~-N ONH ~ ON CONH
R OH
The title ssmpound is obtained ~nalogously to
Example XXXIV by remo~al of the Boc protecting group
~rom the compound from Ex~mple XVII.
~ Li~
TLC-system III: Rf - 0~38
Exam~le LXVII
L-(4-methoxy)phenylal~nyl-~2-R,S-amino-2-~2-~1,3-
dithiolano)]}acetyl-4S-amino-3S-~ydroxy-5-cyclohexylpen-
tanoyl-L-i~oleucine-(2-picolyl)amide-dihydrochloride
Le A 26 914 - 91 -
~` 2Vl~
OCH
~ S ~ ~ ~ * HCl
L * ~ ONH ONH
R,S OH
The title compound is obtained analogously ~o
Example XXXIV by removal of the ~oc protecting group
from the mixture of the compounds from Example XVI and
XVII.
Analvtlcal da~a:
TLC-System lII: Rf = 0.42:
Rt = 0.38
ExamDles LXVIII o LXXVI ~Table 1)
The Examples from Table 1 are prepared analogously
to Example 3 or I by n-propanephosphonic acid anhydride
coupling of the compound from Example XXV with the
corresponding amine compounds (analogously to Example
LII to LIX). The puri~ication and/or 6eparation o~ ~he
stereoisomers i5 csrried out by chromatography ~a~
deccribed in Examples XLIII and XLIV~.
-
Le A 26 914 - 92
~ ~ '
.
..
.. :
'
' 2 ~ g
S Z ~ X ~
U~ ~ ~D 0 0 t~
-- ~
0
C C C C C
i~E3 Ei E 13 E
l l -I ~ ~ 0 U~
t~ ~N ~ U) D ~D tq ~D
~D oD a) o ~ ~ ~
-- _ .
E3 tr~ ~ ~ CO ~ ~I N
O ~ O O O O O
.. .. .. .. .. ..
. . H H ~ H H H
~1 1~ H H H 1--1
H/--1 H H ~--1
X
~ ~* --~
~ Y ~ ~ ~
0
Z _
~ O ~ ~ ~
_, ~ ~e x ~
Le ~ 26914 - 93 -
- -~ 2~8~8
_ _ ~
:~:
r~ I
N IJ~
t~
+
a~ _ _
El E E
.. . ~
H
~ 3
~n _, ,,, ._1 . rl
E E3 E
oo
o ~o
.. ..
U~
`T'N
_ . _ _ . _ _ _
E~ -I ~ O
Q~ ~ ~` ~O `O
:~., .
O O
I
.. ....
I
I
~ I .
t'q
/
~ O
X ~ *~
Z :~ Z
O
.,,1 1 *
U~
.rl O .
~-r1
O ~
C: O L
C~ . _ __
._
_~
..q 6' X ~X
ID X
E~ L~
Le A26 914 - 94 -
~ .
,
:
201~
Examples LXXVII ~o LXXXI (Table 2)
The examples from table 2 are ob~a;ned analogously
~o Example XXXVI by reac~ing ~he Dihydrochlorides from
Example LXVI andlor Example LXVII wi~h ~he corresponding
acids using n-propanephosphonic acid anhydride as
coupling reagen~. The purification is carried ou~ by
chroma~ography.
Ls A ?6 914 - 95 -
2 ~
;
, ~
,~
u~ ~ X
:E: :~
~ ~ ~D O
0 o~
` E9 E; E3
~ ~ C ~ ~ C
U~ _, .,. .,1 ... ...
l l
o
., .
, , _ , ___
O
n~ a~ u~ ~ N
~ O O O O O
Z U~ ~ .. .. .. .. ..
O I I
O
__
*
O C
A y , o
~rl
~ ~n ~--O ~:: L a~
o ~
- ~q ~ ~
:~:
z
4~ o o l l ~
* o
o~ I ~ z~ Q O ~
Z _
N
~ X ~ ~ ~
~ _l ~ ~ X ~
~ 9 ~ ~ X ~
Le A 26 914 - 96 -
: , ., - ;
. .
.
.
,
-
Exam~le LXXXII to LXXXVI (Table 3)
The Examples from table 3 are ~b~ained by n-
propanephosphonic acid anhydride coupling of the
corresponding Boc-protected amino acids (BISSENDORF
BIOCHEMICALS GmbH) with ~he compound from Example LXV
(analogously to Example XVI and XVII) and chromato-
graphic purifica~ion.
~0
Le A 26 914 - 97 -
2 ~
~ ,.~
U~ + + + + +
U~ ~ ~ ~,,
o o~
C~
E El E 1
a~
~ ~ C C ~ ~ C
Z~ u), E~ E E E e
L~
O ~ ~ ~ ~O O`
~; .~
~ ~o o r~ ~ Ln
(~
~ I
~: e ~
Z ~ a~ r~
O
A / ~ ~ o O o o o
X > ~ .. .. .. .. ..
X ~ I
O ~
, ~ -- , .
o~ ~l~qo=l l ~
~ u
.
~ o ~ ~ ~ ~ ~
m _,
n e X X X X X
~ x ~ ~ ~ ~ ~
r~
Le A 26 914 -98 -
.
'
.
Examples LXXXVIl tD XC (Table 4)
The Examples from table 4 are either prepared by
coupling the corresponding acids tExample LXXXVII ~o
IXC) w;Lh the compo~nd from Example LXV using n-propane-
phosphonic acid anhydride or by reacting ~he compound
from Example LXV with 'Z-OSu' in the presence of
trie~hylamine (Example XC). As to s~andard workup all
compounds are purified by chromatography.
The acids are prepared by known synthe~ic me~hods.
The synthesis of ~he a~id used for the prepara~ion of
Example LXXXVII is described in Example 19. The synthe-
sis of ~he acids for Example LXXXVIII and IXC arPpublished elsewhere (LXXXVII: J,J, Plattner et al., J.
Med, Chem. 1988, 31, 2277-2288, IXC: P. Buelmayer at
al., J, Med. Chem, 1988, 31, 1839-1846) ,Z-050' ~N-
(Benzyloxycarbonylo~y)-succinimid] is commercially
available tAldrich Chemicals).
Le A 26 914 - 99 -
.
.,. ....
.~ ~ ~
, ~ C Z
s~ + t I I
:~ -- ~
U~
m , o N ~ tt)
~C ~
03 CO CO
Ql
E~ E~ E~
_
~ ~ '~
~ C
~_, ~3 E3 ` ~
u~ ~ u~
l l ~ ~ ~ ~ ~
O ~U~
~;
, . .
~ O
., q .~
Z U~ _1o 11 o o o
o :~ ~ . `
O U~ ~ .. .. .. ..
l l H
~_ _1 ~ ~
E~ ~ ~
O
Q~ ~o
~ I o ~ ~2y
~ ~ ~ O Z O :C
~O)
~1 G ~ -
.4 ~ X ~ ~
X
Le A 26 914 - 100 -
'
,
: ~:
,
'
,
Explanations of the experimental section:
TLC systems:
Stationary Dhase
Merck prepared TLC plates silica gel 60 F-254, 5 x l0 cm,
layer thickness 0.25 mm, Art. No. 5719.
Mobile phases (in the test as "TLC system")
I : CH2Cl2~MeOH 9:l
II : CH2Cl2t~eOH 95:5
III : NH3/CH2Cl2~MeOH 0.2:9:l
IV : HOAc/C~2Cl2/M~OH 0.2:9:l
V : Glacial acetic acid/n-butanol/H2O l:3:l
VI : EtOAc/n-hexane 2:l
VII : EtOAc/n-hexane 1:3
VIII : EtOAc/n-hexane l:l
IX : CH2Cl2/MeOH 98:2
X : CH2Cl2/MeOH 7:3
XI CH2Cl2
HPLC systems:
HPLC system I : Column Merck Lichrosorb~ RP-8, 250-4,
l0 ~m, Cat. No. 5031B
HPLC system II : Column ~erck Lichrosorb~ RP-18, 250-4,
l0 ~m, Cat. No. 50334
Eluent for systems I and II
A: pH 7.00 phosphate buffer, Merck, Art. No. 9439/H20
1s50
B~ Acetonitrile
Le A 26 914 -101-
. ,. . . ~ .
,
- 2~8~
A/B as 1/l, flow rate: 2 ml/min, isocratic,
Detection: 254 nm
Index_of the abbreviations used
1. General analytical methods
TLC Thin layer chromatography
PTLC Preparative thick layer chromatography
GC Gas chromatography
HPLC High pressure liquid chromatography
CC Column chromatography
NMR Nuclear magnetic resonance spectroscopy
(protons)
MS Mass spectrometry (electron Lmpact ionization)
~+)F~B-MS Fast atom bombardment mass spectrometry,
po~itive ions, matrix substance: m-nitrobenzyl
alcohol
MS-DCI Mass spectrometry, chemical ionization
2. Protectina ~roups
Boc Tert-butoxycarbonyl
Z Benzyloxycarbonyl
DNP Dinitroph~nyl
Fmoc 9-Fluorenylmethoxycarbonyl
OEt Ethyl ester
OMe ~ethyl ester
EtOC Ethoxycarbonyl
It will be understood that the specification and
examples are illustrative but not limitative of the
present invention and that other embodiments within the
spirit and scope of the invention will suggest themselves
to those skilled in the art.
aCLaL2ç_2l~ -102-
