Note: Descriptions are shown in the official language in which they were submitted.
W094/229l0 ~ PCT~S94/032~
21~9~ ~
TTTLE
Cyclic Compounds Useful as Inhibitors of Platelet
Glycoprotein IIb/IIIa
FIF.T,n OF T~F~ INVF NTION
This invention relates to novel cyclic compounds
containing carbocyclic ring systems useful as
antagonists of the platelet glycoprotein IIb/IIIa
complex, to pharmaceutical compositions containing such
cyclic compounds, with or without other therapeutic
agents, and to methods of using these compounds, with or
without other therapeutic agents, for the inhibition of
platelet aggregation, as thrombolytics, and/or for the
treatment of other thromboembolic disorders.
RACKGROUND OF T~F. I~VF.NTIOI~
Activation of platelets and the resulting platelet
aggregation and secretion of factors by the platelets
has been associated with different pathophysiological
conditions including cardiovascular and cerebrovascular
25 thromboembolic disorders, for example, the
thromboembolic disorders associated with unstable
angina, myocardial infarction, transient ischemic
attack, stro~e, atherosclerosis and diabetes. The
contribution of platelets to these disease processes
stems from their ability to form aggregates, or platelet
thrombi, especially in the arterial wall following
injury or plaque rupture.
Platelets are known to play an essential role in
the maintenance of hemostasis and in the pathogenesis of
35 arterial thrombosis. Platelet activation has been shown
to be enhanced during coronary thrombolysis which can
- WO94122910 PCT~S94/032~
2~9~
lead to delayed reperfusion and reocclusion. Clinical
studies with aspirin, ticlopidine and a monoclonal
antibody for platelet glycoprotein IIb/IIIa provide
biochemical evidence for platelet involvement in
unstable angina, early stage of acute myocardial
infarction, transient ischemic attack, cerebral
ischemia, and stroke.
Platelets are activated by a wide variety of
agonists resulting in platelet shape change, secretion
of granular contents and aggregation. Aggregation of
platelets serves to further focus clot formation by
concentrating activated clotting factors in one site.
Several endogenous agonists including adenosine
diphosphate ~ADP), serotonin, arachidonic acid,
thrombin, and collagen, have been identified. Because
of the involvement of several endogenous agonists in
activating platelet function and aggregation, an
inhibitor which acts against all agonists would
represent a more efficacious antiplatelet agent than
currently available antiplatelet drugs, which are
agonist-specific.
Current antiplatelet drugs are effective against
only one type of agonist; these include aspirin, which
acts against arachidonic acid; ticlopidine, which acts
against ADP; thromboxane A2 synthetase inhibitors or
receptor antagonists, which act against thromboxane A2;
and hirudin, which acts against thrombin.
Recently, a common pathway for all known agonists
has been identified, namely platelet glycoprotein
IIb/IIIa complex (GPIIb/IIIa), which is the membrane
protein medlating platelet aggregation. A recent review
of GPIIb/IIIa is provided by Phillips et al. (l99l) Cell
65: 359-362. The development of a GPIIb/IIIa antagonist
represents a promising new approach for antiplatelet
therapy. Recent studies in man with a monoclonal
PCT~S94/03
WO94/22910
~ 215.g~70
antibody for GPIIb/IIIa indicate the antithrombotic
benefit of a GPIIb/IIIa antagonist.
There is presently a need for a GPIIb/IIIa-specific
antiplatelet agent which inhibits the activation and
aggregation of platelets in response to any agonist.
Such an agent should represent a more efficacious
antiplatelet therapy than the currently available
agonist-specific platelet inhibitors.
GPIIb/IIIa does not bind soluble proteins on
unstimulated platelets, but GPIIb/IIIa in activated
platelets is known to bind four soluble adhesive
proteins, namely fibrinogen, von Willebrand factor,
fibronectin, and vitronectin. The binding of fibrinogen
and von Willebrand factor to GPIIb/IIIa causes platelets
to aggregate. The binding of fibrinogen is mediated in
part by the Arg-Gly-Asp (RGD) recognition sequence which
is common to the adhesive proteins that bind GPIIb/IIIa.
Several RGD-containing peptides and related
compounds have been reported which block fibrinogen
binding and prevent the formation of platelet thrombi.
For example, see Cadroy et al. (1989) J. Clin. Invest.
84: 939-944; Klein et al. U.S. Patent 4,952,562, issued
8/28/90; European Patent Application EP 0319506 A;
European Patent Application EP 0422938 A1; European
Patent Application EP 0422937 Al; European Patent
Application EP 0341915 A2; PCT Patent Application WO
89/07609; PCT Patent Application WO 90/02751; PCT Patent
Application WO 91/04247; and European Patent Application
EP 0343085 A1.
In the present invention we use conformationally-
constraining carbocyclic ring systems as templates for
. cyclizing peptides such that they have high affinity and
selectivity for GPIIb/IIIa.
.
~U~Y OF T~F. INVF.NTION
PCT~S94/032
W094/~910
2l~o~a
This invention provides novel cyclic compounds
containing carbocyclic ring systems useful as
antagonists of the platelet glycoprotein IIb/IIIa
complex, pharmaceutical compositions containing such
cyclic compounds, and methods of using these compounds,
alone or in combination with other therapeutic agents,
for the inhibition of platelet aggregation, as
thrombolytics, and/or for the treatment of
thromboembolic disorders.
This invention also relates to combination
products, that is, pharmaceutical compositions
containing the novel cyclic compounds of the invention
in combination with anti-coagulants such as warfarin or
heparin, or anti-platelet agents such as aspirin,
piroxicam or ticlopidine, or thrombin inhibitors such as
boropeptides, hirudin or argatroban, or thrombolytic
agents such as tissue plasminogen activator,
anistreplase, urokinase or streptokinase, or
combinations thereof, to pharmaceutical kits containing
these combination products, and to methods of using
these combination products for the inhibition of
platelet aggregation, as thrombolytics, and/or for the
treatment of thromboembolic disorders.
RRI~F DFSCRIPTION OF T~F FIGURF.S
Fi~ure I. Figure I shows the platelet
deaggregatory and thrombolytic effects of the cyclic
IIb/IIIa antagonist compounds cyclo-(D-AbuNMeArg-Gly-
Asp-Mamb) (Compound A) and cyclo-(D-Val-NMeArgGly-Asp-
Mamb) (Compound B) at varying concentrations on an
already formed platelet-rich clot. The clot was formed
by incubating the platelets with agonists for 30
minutes. The cyclic compounds of the present invention
had a significant lytic effect on the clot, with an ICso
of about 0.0005 mM for Compound A. By comparison, the
. : ' !
W094l22910 PCT~S94/032~
21~7~
linear peptide RGDS was much less effective as a
thrombolytic, even at substantially higher
concentrations.
Figure II. Figure II shows the thrombolytic effect
of the cyclic IIb/IIIa antagonist compounds cyclo-(D-
AbuNMeArg-Gly-Asp-Mamb) (Compound A) and cyclo-(D-Val-
NMeArgGly-Asp-Mamb) (Compound B), and the standard
thrombolytics tissue plasminogen activator (tPA),
urokinase (UK) and streptokinase (SK) on an already
formed platelet-rich clot. The clot was formed by
incubating the platelets with agonists for 30 minutes.
Both Compounds A and B showed a significant thrombolytic
effect as compared to the standard thrombolytics tissue
plasminogen activator, urokinase, and streptokinase.
F;gllre III. Figure III shows the thrombolytic
effect of the cyclic compound cyclo-(D-AbuNMeArg-Gly-
Asp-Mamb) (Compound A) and the standard thrombolytics
tissue plasminogen activator (tPA), urokinase (UK), and
streptokinase (SK), both alone and in combination, on an
already formed platelet-rich clot. The c~ot was formed
by incubating the platelets with agonists for 30
minutes. Compound A showed a significant thrombolytic
effect, providing significant clot lysis at 1.0 uM.
Moreover, Compound A in combination with tissue
plasminogen activator, urokinase, or stréptokinase was
significantly better than Compound A alone, and
significantly better than the additive effects of both
agents administered alone.
F;~l~re IV. Figure IV shows the thrombolytic effect
of the cyclic IIb/IIIa antagonist compound cyclo-(D-Val-
NMeArg-Gly-Asp-Mamb) (Compound B) and the standard
thrombolytics tissue plasminogen activator (tPA),
urokinase (UK) and streptokinase (SK), both alone and in
WO94/22910 PCT~S94/032~
2i~
combination, on an already formed platelet-rich clot.
The clot was formed by incubating the platelets with
agonists for 30 minutes. Compound B showed a
significant thrombolytic effect, providing significantly
better clot lysis than tissue plasminogen activator,
urokinase or streptokinase. Moreover, Compound B in
combination with tissue plasminogen activator, urokinase
or streptokinase was significantly better than Compound
B alone, and significantly better than the additive
effects of both agents.
F;gure V. Figure V shows the thrombolytic effect
of the cyclic compound cyclo-(DVal-NMeArg-Gly-Asp-
MeMamb) (isomer 1; the compound of Example 68) (Compound
C) alone and in combination with the standard
thrombolytics tissue plasminogen activator (tPA),
urokinase (UK) and streptokinase (SK) on an already
formed platelet-rich clot. The clot was formed by
incubating the platelets with agonists for 30 minutes.
Compound C alone showed a significant thrombolytic
effect. In combination with tissue plasminogen
activator, urokinase or streptokinase, a thrombolytic
effect was achieved which was greater than the additive
effect of the agents when administered alone.
F;gure VI. Figure VI shows the thrombolytic effect
of the cyclic compound cyclo-(D-Val-NMeArg-Gly-Asp-
MeMamb) (isomer 2; the compound of Example 68a)
(Compound D) alone and in combination with the standard
thrombolytics tissue plasminogen activator (tPA),
urokinase (UK) and streptokinase (SK) on an already
formed plateletrich clot. The clot was formed by
incubating the platelets with agonists for 30 minutes.
Compound D alone showed a significant thrombolytic
effect. In combination with tissue plasminogen
activator, urokinase or streptokinase, a thrombolytic
W094/22910 215 9 0 7 0 PCT~S94/032~
effect was achieved which was greater than the additive
effect of the agents when administered alone.
Figure VII. Figure VII shows the in vivo
thrombolytic and anti-thrombotic effect of the cyclic
glycoprotein IIb/IIIa compound cyclo-(D-Abu-NMeArg-Gly-
AspMamb) (Compound A), alone or in combination with the
standard thrombolytic streptokinase (SK). The
experiments were carried out using an arterial
thrombosis animal model. Figure VII shows the results of
initial administration as a percentage of clot lysis.
Compound A alone showed good in vivo thrombolytic
efficacy, and the use of Compound A with streptokinase
resulted in an increase in in vivo thrombolytic efficacy
while allowing a significantly lower dose of
streptokinase. This study demonstrated significant
reduction in the dose of streptokinase required to
achieve 100% lysis in vivo when Compound A is
administered along with streptokinase.
F;~ure VIII. Figure VIII a-d shows the results of
administration of Compound A or saline following
streptokinase (SK) or tissue plasminogen activator
(t-PA~ thrombolysis, with the results reported as time
to reocclusion and percentage of reocclusion. The
saline control showed 100% reocclusion, whereas
administration of Compound A resulted in virtually no
reocclusion.
D~TAIT~Fln D~SCRIPTION OF T~F INV~NTION
[1] This invention is directed to novel compounds of
the formula
PCT~S94/03223
WO94/22910
.
2159 ~ ~ K L
/ M
R32 NR2
/
,R2,2 (R23) C~n~ C (R21 ) Rl )
\ R3l ~ ,~,
(I)
or a pharmaceutically acceptable salt or prodrug form
thereof wherein:
R31 is a C6-C14 saturated, partially saturated,
or aromatic carbocyclic ring system
substituted with 0-4 R10 or RlOa
R32 is selected from:
--C (=O) ~;
--C (=S)--
-S(=O)2-;
-S(=O)-;
-P(=z)(zR13)_;
Z is S or O;
n" and n' are independently 0-2;
R1 and R22 are independently selected from the following
groups:
hydrogen,
C1-Cg alkyl substituted with 0-2 Rll;
C2-Cg alkenyl substituted with 0-2 R11;
C2-Cg alkynyl substituted with 0-2 R11;
C3-C10 cycloalkyl substituted with 0-2 R11;
--8--
W094/22910 215 9 ~ 7 0 PCT~S94/032~
.
aryl substituted with 0-2 R12;
a 5-10-membered heterocyclic ring system containing
1-4 heteroatoms independently selected from N, S,
and 0, said heterocyclic ring being substituted
with 0-2 R12;
=0, F, Cl, Br, I, -CF3, -CN, -C02R13~ -C(=o)R13,
-C(=o)N(Rl3)2~ -CH0, -CH20R13, -oC(=o)R13,
-oC(=o)oR13a~ -oR13, -oC(=o)N(R13)2, -NR13C(=o)R13,
NR14c(=o)oRl3a~ -NRl3c(=o)N(Rl3)2
-NR14So2N(Rl3)2~ -NR14So2Rl3a~ -S03H, -So2Rl3
-SR13, S(=o)R13a~ -So2N(Rl3)2~ -N(R13)2,
--NHC(=NH)NHR13~ --C(=NH)NHR13, =NoR13, N02,
-C(=o)NHoR13, -C(=o)NHNR13R13a, -OCH2C02H,
2-(1-morpholino)ethoxy;
R1 and R21 can alternatively join to form a 3-7 membered
carbocyclic ring substituted with 0-2 Rl2;
when n' is 2, R1 or R21 can alternatively be taken
together with R1 or R21 on an adjacent carbon atom
to form a direct bon~, thereby to form a double or
triple bond between said carbon atoms;
R22 and R23 can alternatively join to form a 3-7 membered
carbocyclic ring substituted with 0-2 Rl2;
when n" is 2, R22 or R23 can alternatively be taken
together with R22 or R23 on an adjacent carbon atom
to forn: a direct bond, thereby to form a double or
triple bond between the adjacent carbon atomsi
_g_
WO94122910 PCT~S94/032~
2~9~
Rl and R2, where R2l is H, can alternatively join to form
a 5-8 membered carbocyclic ring substituted with
0-2 Rl2;
Rll is selected from one or more of the following.
~,
=O, F, Cl, Br, I, -CF3, -CN, -Co2Rl3~ -C(=o)Rl3,
-C(=o)N(Rl3)2~ -CHO, -CH20Rl3, -oC(=o)Rl3,
-oc(=o)oRl3a~ -oRl3, -oc(=o)N(Rl3)2, _NRl3C(=o~Rl3,
-NRl4C(=o)oR13a~ -NRl3C(=o)N(Rl3)2
-NRl4So2N(Rl3)2~ -NRl4So2Rl3a~ -SO3H, -So2Rl3
_SRl3, s(=o)Rl3a~ -S02N(Rl3)2, -N(Rl3)2,
-NHC(=NH)NHRl3, -C(=NH)NHRl3, =NoRl3, NO2,
-C (=O) NHoR13, -C (=O) NHNR13R13a, -OCH2C02H,
2-(l-morpholino)ethoxy,
Cl-Cs alkyl, C2-C4 alkenyl, C3-c6 cycloalkyl, C3-C6
cycloalkylmethyl, C2-C6 alkoxyalkyl, C3-C6
cycloalkoxy, Cl-C4 alkyl (alkyl being substituted
with 1-5 groups selected independently from:
-NRl3Rl4, -CF3, NO2, -So2Rl3a~ or -S(=o)Rl3a),
aryl substituted with 0-2 Rl2,
a 5-lO-membered heterocyclic ring system containing
1-4 heteroatoms independently selected from N, S,
and O, said heterocyclic ring being substituted
with 0-2 Rl2;
Rl2 is selected from one or more of the following:
phenyl, benzyl, phenethyl, phenoxy, benzyloxy,
halogen, hydroxy, nitro, cyano, Cl-Cs alkyl, C3-C6
cycloalkyl, C3-C6 cycloalkylmethyl, C7-Clo
arylalkyl, Cl-Cs alkoxy, -Co2Rl3~ -C(=o)NHoRl3a,
-C(=o)NHN(Rl3)2, =NoRl3, -B(R34)(R35), C3-C6
--10--
WO94/22910 PCT~S94/032~
215907~
.
cycloalkoxy, -oC(=o)R13, -C(=o)R13,-oC(=o)oRl3a/
-oR13, -(Cl-C4 alkyl)-ORl3, -N(R13)2,
OC( o)N(R13)2, -NR13C(=o)R13, -NRl3C(=o)oRl3a
-NRl3C(=o)N(Rl3)2, -NR13So2N(Rl3)2, -NR13So2Rl3
-SO3H, -So2Rl3a~ -S(=o)R13a, -SR13, -So2N(Rl3)2~
C2-C6 alkoxyalkyl, methylenedioxy, ethylenedioxy,
Cl-C4 haloalkyl, Cl-C4 haloalkoxy, Cl-C4
alkylcarbonyloxy, Cl-C4 alkylcarbonyl, Cl-C4
alkylcarbonylamino, -OCH2CO2H,
2-(1-morpholino)ethoxy, Cl-C4 alkyl (alkyl being
substituted with -N(R13)2, -CF3, NO2, or
s(=o)Rl3a);
R13 is selected independently from: H, Cl-Clo alkyl,
C3-Clo cycloalkyl, C4-C12 alkylcycloalkyl, aryl,
-(Cl-Clo alkyl)aryl, or C3-Clo alkoxyalkyl;
R13a is Cl-Clo alkyl, C3-Clo cycloalkyl, C4-C12
alkylcycloalkyl, aryl, -(Cl-Clo alkyl)aryl, or
C3-Clo alkoxyalkyl;
when two R13 groups are bonded to a single N, said R13
groups may alternatively be taken together to form
-(CH2)2-s- or -(cH2)o(cH2)-;
R14 is OH, H, Cl-C4 alkyl, or benzyl;
R21 and R23 are independently selected from:
hydrogen;
Cl-C4 alkyl, optionally substituted with 1-6
halogen;
benzyl;
R2 is H or Cl-Cg alkyl;
PCT~S94/032~
R10 and R10a are selected independently from one or more
of the following:
phenyl, benzyl, phenethyl, phenoxy, benzyloxy,
halogen, hydroxy, nitro, cyano, Cl-C5 alkyl, C3-C6
cycloalkyl, C3-C6 cycloalkylmethyl, C7-C1o
arylalkyl, C1-Cs alkoxy, -Co2Rl3~ -C(=o)N(Rl3)2
-C(=o)NHoR13a, -C(=o)NHN(Rl3)2~ =NoR13,
-B(R34)~R35), C3-C6 cycloalkoxy, -oC(=o)R13,
-C(=o)R13,-oC(=o)oRl3a~ -oR13, -(C1-C4 alkyl)-OR13,
-N(R13)2, _oc(=o)N(Rl3)2, -NRl3c(=o)Rl3
NR13C(=o)oR13a~ -NR13C(=o)N(R13)2,
-NR13So2N(R13)2, -NR13So2Rl3a~ -S03H, -So2R13a,
-S(=o)R13a, -SR13, -So2N(Rl3)2~ C2-C6 alkoxyalkyl,
methylenedioxy, ethylenedioxy, C1-C4 haloalkyl
(including -CVFw where v = 1 to 3 and w = 1 to
(2v+1)), C1-C4 haloalkoxy, C1-C4 alkylcarbonyloxy,
C1-C4 alkylcarbonyl, C1-C4 alkylcarbonylamino,
-0CH2C02H, 2-(1-morpholino)ethoxy, C1-C4 alkyl
(alkyl being substituted with -N(R13)2, -CF3, N02,
or -S(=o)R13a);
J is ~Ala or an L-isomer or D-isomer amino acid of
structure -N(R3)C(R4)(R5)C(=o)-, wherein:
R3 is H or C1-Cg alkyl;
R4 is H or C1-C3 alkyl;
R5 is selected from:
hydrogen;
C1-Cg alkyl substituted with 0-2 R11;
C2-Cg alkenyl substituted with 0-2 R11;
C2-Cg alkynyl substituted with 0-2 R11;
C3-C1o cycloalkyl substituted with 0-2 Rl1;
-12-
WO94/229l0 PCT~S94/032~
~ ~15gO7~
aryl substituted with 0-2 R12;
a 5-10-membered heterocycllc ring system
containing 1-4 heteroatoms independently
selected from N, S, or O, said heterocyclic
ring being substituted with 0-2 R12;
=O, F, Cl, Br, I, -CF3, -CN, -Co2Rl3~
-C(=o)R13, -C(=o)N(R13)2, -CHO, -CH20R13,
--OC (=O) R13, --OC (=O) oR13a, --OR13,
-oC(=o)N(R13)2, -NR13c(_o)Rl3
-NR14C(=o)oR13a~ -NR13C(=o)N(Rl3)2~
-NR14So2N(Rl3)2~ -NR14So2Rl3a~ -SO3H,
sO2Rl3a~ _SR13, -s(=o)Rl3a~ -So2N(Rl3)2
-N(R13)2, -NHC(=NH)NHR13, -C(=NH)NHR13,
=NoR13, N02, -C (=O) NHoR13, -C (=O) NHNR13R13a,
=NoR13, -B(R34)(R35), -OCH2CO2H,
2-(l-morpholino)ethoxy, -SC(-NH)NHR13, N3,
-Si(CH3)3, (C1-Cs alkyl)NHR16;
-(Co-C6 alkyl)X;
--( CH2 ) q~CH2 ) q~X
, where q is
independently 0,1;
--CH2 {} CH2X
-(CH2)mS(O)p.(CH2)2X, where m = 1,2 and p' =
0-2;
wherein X is defined below; and
PCT~S94/032
WO94/22910
215~~
R3 and R4 may also be taken together to form
(CH2) nX
I
--CH2CHCH2--, where
~NR13
-NH-C
n = 0,1 and X is N(Rl3)R13;
R3 and R5 can alternatively be taken together to form
-(CH2)t- or -CH2S(O)p~C(CH3)2-, where t = 2-4 and p'
= 0-2; or
0 R4 and R5 can alternatively be taken together to form
-(CH2)U-, where u = 2-5;
R16 is selected from:
an amine protecting group
1-2 amino acids;
1-2 amino acids substituted with an amine
protecting group;
0 ~ is a D-isomer or L-isomer amino acid of structure
-N(R6)CH(R7)C(=o)-, wherein:
R6 is H or C1-Cg alkyl;
R7 is selected from:
-(Cl-C7 alkyl)X;
--( CH2 ) q~
(CH2)q~X wherein each q is
independently 0-2 and substitution on the phenyl
is at the 3 or 4 position;
-14-
W094l22910 215 9 Q 7 Q PCT~S94/032~
-(CH2)q ~
\___/ (CH2)q-X wherein each q is
independently 0-2 and substitution on the
cyclohexyl is at the 3 or 4 position;
-(C1-C6 alkyl) \ ~
NH
~ )0-3
~(CH2)mO-(C1-C4 alkyl)-X, where m = 1 or 2;
~(CH2)mS(O)p.-(C1-C4 alkyl)-X, where m = 1 or 2
and p' = 0-2; and
X is selected from:
~ NR13
-NH-C
N~Rl3)Rl3; _N(R13)R13; -C(=NH)(NH2);
-SC(=NH)-NH2; -NH-C(=NH)(NHCN);
-NH-C(=NCN)(NH2); -NH-C(=N-oR13)(NH2);
R6 and R7 can alternatively be taken together to form
(CH2) nX
~(CH2)qCH(CH2)q , wherein each q is independently
1 or 2 and wherein
n = 0 or 1 and X is -NH2 or
~ NRl 3
-NH-C
N(R13)Rl3
-15-
WO94/22910 PCT~S94/03223
~9~
L is -Y(CH2)VC(=O)-, wherein:
Y is NH, N(Cl-C3 alkyl), o, or S; and v = 1 or 2;
M is a D-isomer or L-isomer amino acid of structure
-NR17-CH-c(=o)
(CH(R4))q~
10 R8 , wherein:
q' is 0-2;
R17 is H, Cl-C3 alkyl;
R8 is selected from:
-Co2Rl3~-so3Rl3~ -So2NHR14, -B(R34)(R35), -NHSO2CF3,
-CONHNHSO2CF3, -Po(oRl3)2~ -Po(oR13)R13,
-SO2NH-heteroaryl (said heteroaryl being
5-10-membered and having 1-4 heteroatoms selected
independently from N, S, or O) , -SO2NH-heteroaryl
(said heteroaryl being 5-10-membered and having 1-4
heteroatoms selected independently from N, S, or
O), --So2NHCoRl3, --CoNHSo2Rl3a, -CH2CoNHSo2Rl3a,
-NHSo2NHcoRl3a~ -NHCoNHS02R13a, -S02NHCoNHR13,
--C02R13b;
R34 and R35 are independently selected from:
-OH,
-F,
-N(R13)2, or
Cl-Cg-alkoxy;
R34 and R35 can alternatively be taken together form:
-16-
W094/22910 PCT~S94/032~
2ls~07n
.
a cyclic boron ester where said chain or ring
contains from 2 to 20 carbon atoms and,
optionally, 1-4 heteroatoms independently
selected from N, S, or O;
a divalent cyclic boron amide where said chain or
t ring contains from 2 to 20 carbon atoms and,
optionally, 1-4 heteroatoms independently
selected from N, S, or O;
a cyclic boron amide-ester where said chain or ring
contains from 2 to 20 carbon atoms and,
optionally, 1-4 heteroatoms independently
selected from N, S, or O;
R13b is selected from:
(a) C1-Cg alkyl;
(b) C2-Cg alkenyl;
(c) C2-Cg alkynyl;
(d) C3-Cg cycloalkyl;
(e) C1-Cg alkyl substituted with
(i) aryl, optionally substi~uted with 1-2
substituents independently selected
from halogen, phenyl, C1-Cs alkyl,
C1-Cs alkoxy, NO2, -S(O)0-2(C1-C5
alkyl), OH, N(R13)2, C02R13, CoN(Rl3)2
or -CVFw where v = 1 to 3 and w = 1 to
(2v+1);
(ii) C3-Cg cycloalkyl;
(iii)
~(CH2)w
l40
(f) aryl, optionally substituted with 1-2
substituents independently selected from
WO94/22910 PCT~S94/03223
s~ S ~
halogen, phenyl, C1-Cs alkyl, Cl-C5 alkoxy,
NO2, -s(o)o-2(cl-c5 alkyl), OH, N(R13)2,
C02R13, CON(R13)2 or -CVFw where v = 1 to 3
and w = 1 to ~2v+1)i
(g) C2-Cg alkyl,`alkenyl or alkynyl; substituted
with 1-2 substituents independently
selected from C1-C4 alkyl, C3-Cg cycloalkyl,
C1-Cs alkoxy, phenoxy, benzyloxy, halogen,
NO2, CN, C02R13, CON(R13)2, N(R36)CoR36,
morpholino, 2-(1-morpholino)ethoxy, N~R13)2,
N+(R13)3, OCOCH3, CF3, S(0)o-2Rl3a;
(h) CH(R36)oR38;
(i) CH(R36)oc(=o)R37;
(j) CH(R36)oC(=o)oR38;
(k) CH(R36)oC(=o)N(R37)2;
(1) CH(R36)N(R36)c(=o)R36;
(m) CH(R36)Co2R37;
(n) CH(R36)CON(R13)2;
(o) CH(R36)N(R13)2;
(q)
,1~
O O
R39
(r)
o
OJ~o
~ R39
(s)
-CH(R36)o ~;
O
(t)
-18-
WO94/22910 21~ 9 0~ 0 PCT~S94/032~
.
<o~oo
,~
(u)
~;
R36 is selected independently from: H, C1-Cg alkyl,
C3-Clo cycloalkyl, phenyl, or benzyl;
R37 is selected from:
(a) H;
(b) Cl-Cg alkyl or C3-Cg cycloalkyl, said alkyl or
cycloalkyl being substituted with 1-2 groups
independently selected from:
(i) Cl-C~ alkyl;
(ii) C3-Cg cycloalkyl;
(iii) Cl-Cs alkoxy;
(iv) aryl substituted with 0-2 groups
independently selected from: halogen, phenyl,
Cl-C6 alkyl, Cl-C6 alkoxy, NO2, -S(Cl-Cs
alkyl), -SO(Cl-Cs alkyl), -SO2(Cl-Cs alkyl),
-OH, -N(R13)2, -Co2Rl3r -C(=o)N(Rl3)2r or -CVFw
where v = 1 to 3 and w = 1 to (2v+1);
(c) aryl substituted with 0-2 groups independently
selected from: halogen, phenyl, C1-C6 alkyl,
Cl-C6 alkoxy, NO2, -S(Cl-Cs alkyl), -SO(Cl-Cs
alkyl), -SO2(Cl-Cs alkyl), -OH, -N(R13)2,
-Co2Rl3r -C(-o)N(Rl3)2~ or -CVFw where v = 1 to
3 and w = 1 to (2v+1);
.
R38 is selected from:
-19-
WO94t~910 PCT~S94/032~
~50rlQ ~
(a) Cl-Cg alkyl or C3-Cg cycloalkyl, said alkyl or
cycloalkyl being substituted with 1-2 groups
independently selected from:
(i) Cl-C4 alkyl;
(ii) C3-Cg cycloalkyl;
(iii) Cl-Cs alkoxy;
(iv) aryl substituted with 0-2 groups
independently selected from: halogen, phenyl,
Cl-C6 alkyl, Cl-C6 alkoxy, NO2, -S(Cl-Cs
alkyl), -SO(Cl-Cs alkyl), -SO2(Cl-C5 alkyl),
-OH, -N(R13)2, -Co2Rl3~ -C(=o)N(Rl3)2~ or -CVFw
where v = 1 to 3 and w = 1 to (2v+1);
(b) aryl substituted with 0-2 groups independently
selected from: halogen, phenyl, Cl-C6 alkyl,
Cl-C6 alkoxy, NO2, -S(Cl-Cs alkyl), -SO(Cl-Cs
alkyl), -SO2(Cl-Cs alkyl), -OH, -N(R13)2,
-Co2Rl3~ -C(=o)N(Rl3)2~ or -CVFw where v = 1 to
3 and w = 1 to (2v+1);
R39 is selected from:
(a) H
(b) Cl-Cg alkyl or C3-Cg cycloalkyl, said alkyl or
cycloalkyl being substituted with 1-2 groups
independently selected from:
(i) Cl-C6 alkyl;
(ii) Cl-C6 alkoxy;
(iii) aryl substituted with 0-2 groups
independently selected from: halogen, phenyl,
Cl-C6 alkyl, Cl-C6 alkoxy, NO2, -S(Cl-Cs
alkyl), -SO(Cl-Cs alkyl), -SO2(Cl-Cs alkyl),
-OH, -N(R13)2, -Co2Rl3~ -C(=o)N(Rl3)2r or -CVFw
where v = 1 to 3 and w = 1 to (2v+1);
(c) aryl substituted with 0-2 groups independently
selected from: halogen, phenyl, Cl-C6 alkyl,
Cl-C6 alkoxy, NO2, -S(Cl-Cs alkyl), -SO(Cl-Cs
alkyl), -SO2(Cl-Cs alkyl), -OH, -N(R13)2,
-20-
W094l22910 PCT~S94/032~
215907~
.
-Co2R13, -C(=o)N(R13)2, or -CVFw where v = 1 to
3 and w = 1 to ~2v+1);
R40 is selected from: H, Cl-Cs alkyl, or benzyl;
provided that at least one of the following conditions
is met:
(1) R32 is not -C(=O)-; or
(2) p' is not 0; or
(3) q' is not 0; or
(4) q is not 0-1; or
(5) X is -NH-C(=NH)NHCN, -NH-C(=NCN)(NH2) or
-NH-C(=NoRl3)NH2; or
(5) R8 is -B(R34)(R35) or -Co2Rl3b.
The present invention includes the use of the above
described compounds (wherein the above conditions (l)-
(5) are not required) in combination with one or more
additional therapeutic agents for the inhibition of
platelet aggregation, as thrombolytics, and/or for the
treatment of thromboembolic disorders, wherein the
additional therapeutic agent is selected from:
anti-coagulants such as warfarin or heparin, or anti-
platelet agents such as aspirin, piroxicam or
ticlopidine; thrombin inhibitors such as boropeptides,
hirudin or argatroban; or thrombolytic agents such as
tissue plasminogen activator, anistreplase, urokinase or
streptokinase.
[2] The present invention includes those compounds
above wherein:
R31 is bonded to (C(R23)R22) n" and (C(R2l)Rl) n~ at 2
different atoms on said carbocyclic ring.
-21-
PCT~S94/03223
WO94/22910
gO~a
[3] Included in the present invention are those
compounds above, wherein:
n" is 0 and n' is 0;
n" is 0 and n' is 1;
n" is 0 and n' is 2;
n" is 1 and n' is 0;
n" is 1 and n' is 1;
n" is 1 and n' is 2;
n" is 2 and n' is 0;
n" is 2 and n' is 1; or
n" is 2 and n' is 2.
[4] Included in the present invention are those
compounds of formula (I) above wherein R6 is methyl,
ethyl, or propyl.
[5] This invention includes those compounds above of
the formula:
K - L
J M
R32 NR2
(R22 tR23~ /(R21) R1)
R31
(I)
or a pharmaceutically acceptable salt or prodrug form
thereof wherein:
R31 is a C6-C14 saturated, partially
saturated, or aromatic carbocyclic ring
system substituted with 0-4 R10 or R10a;
R32 is selected from:
W094l22910 2 1~ ~ ~ 7 ~ PCT~S94/03223
--C (=O)--;
--C (=S)--
-S(=O)2-;
Z is S or O;
n" and n' are independently 0-2;
R1 and R22 are independently selected from the following
groups:
hydrogen,
C1-C8 alkyl substituted with 0-2 Rll,
C2-Cg alkenyl substituted with 0-2 Rll,
C2-Cg alkynyl substituted with 0-2 R1l,
C3-Cg cycloalkyl substituted with 0-2 Rll,
C6-C1o bicycloalkyl substituted with 0-2 R1l,
aryl ~ubstituted with 0-2 R12,
a 5-10-membered heterocyclic ring system containing
1-4 heteroatoms independently selected from N, S,
or O, said heterocyclic ring being substituted with
0-2 R12;
=O, F, Cl, Br, I, -CF3, -CN, -Co2R13, -C(=o)R13,
-C(=o)N(R13)2, -CHO, -CH20R13, -oc~=o)Rl3~
-oC(=o)oRl3a~ -oR13, -0C(=o)N(R13)2, -NR13C(=o)R13,
NRl4c(=o)oRl3a~ -NRl3c(=o)N(Rl3)2
-NR14So2N(Rl3)2~ -NR14So2Rl3a~ -SO3H, -So2Rl3
-SR13, -S(=o)R13a, -So2N(Rl3)2~ -CH2N(R13)2,
- --N(R13)2, --NHC(=NH)NHR13, --C(=NH)NHR13, N02;
1 and R2l can alternatively join to form a 5-7 membered
carbocyclic ring substituted with 0-2 R12;
-23-
W094122910 PCT~S94tO32~
2`~59Q~ ~
when n' is 2, Rl or R2l can alternatively be taken
together with Rl or R2l on an adjacent carbon atom
to form a direct bond, thereby to form a double or
triple bond between said carbon atomsi
~
R22 and R23 can alternatively join to form a 3-7 membered
carbocyclic ring substituted with 0-2 Rl2;
when n" is 2, R22 or R23 can alternatively be taken
together with R22 or R23 on an adjacent carbon atom
to form a direct bond, thereby to form a double or
triple bond between said carbon atoms;
(
Rl and R2, where R2l is H, can alternatively join to form
a 5-8 membered carbocyclic ring substituted with
0-2 R12;
Rll is selected from one or more of the following:
=0, F, Cl, Br, I, -CF3, -CN, -Co2Rl3~ -C(=o)Rl3,
-C(=o)N(Rl3)2~ -CHO, -CH20Rl3, -oC(=o)Rl3,
-oc(=o)oRl3a~ -oRl3, _oc(=o)N(Rl3)2, _NRl3C(=o~Rl3,
NRl4c(=O)oRl3a~ -NRl3C(=o)N(Rl3)
-NRl4So2N(Rl3)2~ -NRl4So2Rl3a~ -S03H, -So2Rl3
_SRl3, s(=O)Rl3a~ -S02N(Rl3)2, -CH2N(Rl3)2~
-N(Rl3)2, -NHC(=NH)NHRl3, -C(=NH)NHRl3, =NoRl3,
N02;
.
Cl-Cs alkyl, C2-C4 alkenyl, C3-C6 cycloalkyl, C3-C6
cycloalkylmethyl, C2-C6 alkoxyalkyl, Cl-C4 alkyl
(substituted with -NRl3Rl4, -CF3, N02, -So2Rl3~ or
-S(=o)Rl3a)
aryl substituted with 0-2 Rl2,
-24-
WO94/22910 PCT~S94/032~
2la sn7~
a 5-10-membered heterocyclic ring system containing
1-4 heteroatoms independently selected from N, S,
or O, said heterocyclic ring being substituted with
0-2 R12;
R12 is selected from one or more of the following:
phenyl, benzyl, phenethyl, phenoxy, benzyloxy,
halogen, hydroxy, nitro, cyano, Cl-c5 alkyl, C3-C6
cycloalkyl, C3-C6 cycloalkylmethyl, C7-C1o
arylalkyl, C1-Cs alkoxy, -Co2Rl3~ -C(=o)NHoR13a,
-C(=o)NHN(Rl3)2~ =NoR13, -B(R34)(R35), C3-C6
cycloalkoxy, -oC(=o)R13, -C(=o)R13,-oc(=o)oRl3
-oR13, -(C1-C4 alkyl)-OR13, -N(R13)2,
-0C(=o)N(R13)2, -NR13C(=o)R13, -NR13C(=o)oR13
-NR13C(=o)N(Rl3)2~ -NR13So2N(Rl3)2~ -NR13So2Rl3
-S03H, -So2Rl3a~ -S(=o)R13a, -SR13, -S02N(R13)2,
C2-C6 alkoxyalkyl, methylenedioxy, ethylenedioxy,
C1-C4 haloalkyl, C1-C4 haloalkoxy, C1-C4
alkylcarbonyloxy, C1-C4 alkylcarbonyl, C1-C4
alkylcarbonylamino, -OCH2CO2H,
2-(1-morpholino)ethoxy, Cl-C4 alkyl (alkyl being
substituted with -N(R13)2, -CF3, NO2, or
_s(=o)R13a);
R13 is selected independently from: H, C1-C1o alkyl,
C3-C1o cycloalkyl, C4-C12 alkylcycloalkyl, aryl,
-(C1-C1o alkyl)aryl, or C3-C1o alkoxyalkyl;
30 R13a is C1-C1o alkyl, C3-clo cycloalkyl, C4-C12
alkylcycloalkyl, aryl, -(C1-C1o alkyl)aryl, or
C3-C1o alkoxyalkyl;
when two R13 groups are bonded to a single N, said R13
groups may alternatively be taken together to form
-(CH2)2-s- or -(CH2)O(CH2)-;
WO94/22910 PCT~S94/032~
Z,35g~
R14 is OH, H, C1-C4 alkyl, or benzyl;
R21 and R23 are independently selected from:
hydrogen;
C1-C4 alkyl, optionally substituted with 1-6
halogen;
benzyl;
R2 is H or C1-Cg alkyl;
R10 and Rl0a are selected independently from one or more
of the following:
phenyl, benzyl, phenethyl, phenoxy, benzyloxy,
halogen, hydroxy, nitro, cyano, C1-C5 alkyl, C3-C6
cycloalkyl, C3-C6 cycloalkylmethyl, C7-C1o
arylalkyl, C1-Cs alkoxy, -Co2Rl3~ -C(=o)NHoRl3a,
-C(-o)NHN(R13)2, =NoR13, -B(R34)(R35), C3-C6
cycloalkoxy, -oc(=o)Rl3~ -c(=o)Rl3,-oc(=o)oRl3
-oR13, -(C1-C4 alkyl)-OR13, -N(R13)2,
OC( o)N(R13)2, -NR13C(=o)R13~ -NR13C(=o)oR13a
-NR13C(=o)N(R13)2, -NR13So2N(Rl3)2~ -NR13So2Rl3
-SO3H, -So2Rl3a~ -S(=o)R13a, -SR13, -So2N(Rl3)2~
C2-C6 alkoxyalkyl, methylenedioxy, ethylenedioxy,
C1-C4 haloalkyl, Cl-C4 haloalkoxy, C1-C4
alkylcarbonyloxy, C1-C4 alkylcarbonyl, C1-C4
alkylcarbonylamino, -OCH2CO2H,
2-(1-morpholino)ethoxy, Cl-C4 alkyl (alkyl being
substituted with -N(R13)2, -CF3, NO2, or
-S(=o)R13a);
J is ~Ala or an L-isomer or D-isomer amino acid of
structure -N(R3)C(R4)(R5)C(=o)-, wherein:
-26-
WO94/~910 PCT~S94/032~
~ 21~7~
R3 is H or CH3;
R4 is H or C1-C3 alkyl;
R5 is H, C1-Cg alkyl, C3-C6 cycloalkyl, C3-C6
cycloalkylmethyl, C1-C6 cycloalkylethyl, phenyl,
phenylmethyl, CH2OH, CH2SH, CH2OCH3, CH2SCH3,
CH2CH2SCH3, (CH2)~NH2, (cH2)sNHc(=NH)(NH2)r
(CH2)~NHR16, where s - 3-5;
R3 and R5 can alternatively be taken together to form
-(CH2)t- (t = 2-4) or -CH2SC(CH3)2-; or
R4 and R5 can alternatively be taken together to form
-(CH2)U-, where u = 2-5;
R16 is selected from:
an amine protecting group;
1-2 amino acids;
1-2 amino acids subtituted with an amine
protecting group;
~ is a D-isomer or L-isomer amino acid of structure
-N(R6)CH(R7)C(=o)-, wherein:
R6 is H or C1-Cg alkyl;
R7 is selected from:
-(Cl-C7 alkyl)X;
(CH2) q~
(CH2)q-X wherein each q is
-27-
PCT~S94/032
W094/22910
2~59~
independently 0-2 and substitution on the phenyl
is at the 3 or 4 position;
r~
--( CH2 ) q <
~___/ (CH2):q~X, wherein each q is
independently 0-2 and substitution on the
cyclohexyl is at the 3 or 4 position;
-(Cl-C6 alkyl) \ ~
NH
~ Jo 3
~(CH2)mO-(C1-C4 alkyl)-X, where m = 1 or 2;
~(CH2)mS-(Cl-C4 alkyl)-X, where m = 1 or 2; and
X is selected from:
--NH--C (=NH) (NH2), --NHR13, --C (=NH) (NH2),
--SC (NH)--NH2;
R6 and R7 can alternatively be taken together to form
(CH2) nX
2 0 -CH2CHCH2-, where
n = 0 or 1 and X is -NH2 or -NH-C(=NH) (NH2);
L is -Y (CH2) vC (=O) -, wherein:
Y is NH, N(C1-C3 alkyl), 0, or S; and v = 1 or 2;
M is a D-isomer or L-isomer amino acid of structure
-28-
WO9412291~ PCT~S94/032~
21~9070
-NR17-cH-c(=o)
(CH(R4))~,
R8 , wherein:
q' is 0-2;
~r
Rl7 is H, C1-C3 alkyl;
R8 is selected from:
-Co2Rl3~-so3Rl3~ -So2NHRl4~ -B(R34)(R35), -NHSO2CF3,
-CONHNHSO2CF3, -Po(oRl3)2~ -Po(oR13)R13,
-SO2NH-heteroaryl (said heteroaryl being
5-10-membered and having 1-4 heteroatoms selected
independently from N, S, or O) , -SO2NH-heteroaryl
~said heteroaryl being 5-10-membered and having 1-4
heteroatoms selected independently from N, S, or
O), -So2NHcoRl3~ -CoNHso2Rl3a~ -CH2CoNHso2Rl3
-NHSo2NHCoRl3a~ -NHCoNHSo2Rl3a~ -So2NHCoNHRl3
-Co2Rl3b;
R34 and R35 are independently selected from:
-OH,
-F,
_NR13R14, or
Cl-Cg-alkoxy;
25 R34 and R35 can alternatively be taken together form:
a cyclic boron ester where said chain or ring
contains from 2 to 20 carbon atoms and,
optionally, 1-4 heteroatoms independently
selected from N, S, or o;
a divalent cyclic boron amide where said chain or
ring contains from 2 to 20 carbon atoms and,
optionally, 1-4 heteroatoms independently
selected from N, S, or o;
-29-
W094/22910 PCT~S94/032~
~9~
a cyclic boron amide-ester where said chain or ring
contains from 2 to 20 carbon atoms and,
optionally, 1-4 heteroatoms independently
selected from N, S, or O;
S
R13b is selected from:
(a) C1-Cg alkyl;
(b~ C2-Cg alkenyl;
(c) C2-Cg alkynyl;
(d) C3-Cg cycloalkyl;
(e) Cl-Cg alkyl substituted with
(i) aryl, optionally substituted with 1-2
substituents independently selected
from halogen, phenyl, Cl-Cs alkyl,
Cl-Cs alkoxy, NO2, -S(O)0-2(cl-c5
alkyl), OH, N(R13)2, C02R13, CoN(Rl3)2
or -CVFw where v = 1 to 3 and w - 1 to
(2v+1);
(ii) C3-Cg cycloalkyl;
(iii)
~(CH2)04
R40
. (f) aryl, optionally substituted with 1-2
substituents independently selected from
halogen, phenyl, Cl-Cs alkyl, Cl-Cs alkoxy,
NO2, -S(O)0-2(cl-c5 alkyl), OH, N(R13)2,
C02Rl3, CON(R13)2 or -CVFw where v = 1 to 3
and w = 1 to (2v+1);
(g) C2-Cg alkyl, alkenyl or alkynyl; substituted
with 1-2 substituents independently
selected from Cl-C4 alkyl, C3-Cg cycloalkyl,
Cl-Cs alkoxy, phenoxy, benzyloxy, halogen,
-30-
W094l22910 2 I 5 9 ~ 7 ~ PCT~S94/03223
NO2, CN, C02R13, CON(R13)2, N(R36)CoR36,
morpholino, 2-(1-morpholino)ethoxy, N(R13)2,
N+(R13)3, OCOCH3, CF3, S(0)o-2Rl3
~ (h) CH(R36)oR38;
(i) CH(R36)oC(=o)R37;
(j) CH(R36)oc(=o)oR38;
(k) CH(R36)oc(=o)N(R37)2;
(1) CH(R36)N(R36)C(=o)R36;
(m) CH(R36)Co2R37;
(n) CH(R36)CON(R13)2;
(o) CH(R36)N(R13)2;
(q)
,1~
O O
R39
(r)
,1~
O O
~ ~ R39;
(s)
-CH(R36)o~;
(t)
~0
(u)
-31-
W094/22910 PCT~S94/03223
s~ ~j 9
R36 is selected independently from: H, Cl-Cg alkyl,
C3-Clo cycloalkyl, phenyl, or benzyl;
R37 is selected from:
(a) H;
(b) Cl-Cg alkyl or C3-Cg cycloalkyl, said alkyl or
cycloalkyl being substituted with 1-2 groups
independently selected from:
(i) Cl-C4 alkyl;
(ii) C3-Cg cycloalkyl;
(iii) Cl-Cs alkoxy;
(iv) aryl substituted with 0-2 groups
independently selected from: halogen, phenyl,
Cl-C6 alkyl, Cl-C6 alkoxy, NO2, -S(Cl-Cs
alkyl), -SO(C1-Cs alkyl), -SO2(Cl-Cs alkyl),
-OH, -N(Rl3)2, -Co2Rl3~ -C(=o)N(Rl3)2~ or -CVFw
where v = 1 to 3 and w = 1 to (2v+1);
(c) aryl substituted with 0-2 groups independently
selected from: halogen, phenyl, C1-C6 alkyl,
Cl-C6 alkoxy, NO2, -S(Cl-Cs alkyl), -SOtCl-Cs
alkyl), -SO2(Cl-Cs alkyl), -OH, -N(R13)2,
-Co2Rl3~ -C(=o)N(R13)2, or -CVFw where v = 1 to
3 and w = 1 to (2v+1);
R38 is selected from:
(a) Cl-Cg alkyl or C3-Cg cycloalkyl, said alkyl or
cycloalkyl being substituted with 1-2 groups
independently selected from:
(i) Cl-C4 alkyl;
(ii) C3-Cg cycloalkyl;
(iii) Cl-Cs alkoxy;
(iv) aryl substituted with 0-2 groups
independently selected from: halogen, phenyl,
Cl-C6 alkyl, Cl-C6 alkoxy, NO2, -S(Cl-Cs
alkyl), -SO(Cl-Cs alkyl), -SO2(Cl-Cs alkyl),
-32-
WO94/22910 2 ~ 5 9 ~ 7 ~ PCT~S94/03223
-OH, -N(Rl3)2, -Co2Rl3~ -C(=o)N(Rl3)2, or -CVFw
where v = l to 3 and w = l to (2v+l);
(b) aryl substituted with 0-2 groups independently
selected from: halogen, phenyl, Cl-C6 alkyl,
Cl-C6 alkoxy, NO2, -S(Cl-Cs alkyl), -SO(Cl-Cs
alkyl), -SO2(Cl-Cs alkyl), -OH, -N(Rl3)2,
-Co2Rl3l -C(=o)N(Rl3)2, or -CVFw where v = l to
3 and w = l to (2v+l);
R39 is selected from:
(a) H
(b) Cl-Cg alkyl or C3-Cg cycloalkyl, said alkyl or
cycloalkyl being substituted with 1-2 groups
independently selected from:
(i) Cl-C6 alkyl;
(ii) Cl-C6 alkoxy;
(iii) aryl substituted with 0-2 groups
independently selected from: halogen, phenyl,
Cl-C6 alkyl, Cl-C6 alkoxy, NO2, -S(Cl-Cs
alkyl), -SO(Cl-Cs alkyl), -SO2(Cl-Cs alkyl),
-OH, -N(Rl3)2, -Co2Rl3~ -C(=o)N(Rl3)2~ or -CVFw
where v = l to 3 and w = l to (2v+l);
(c) aryl substituted with 0-2 groups independently
selected from: halogen, phenyl, Cl-C6 alkyl,
Cl-C6 alkoxy, NO2, -S(Cl-Cs alkyl), -SO(Cl-Cs
alkyl), -SO2(Cl-Cs alkyl), -OH, -N(Rl3)2,
-Co2Rl3~ -C(=o)N(Rl3)2~ or -CVFw where v = l to
3 and w = l to (2v+l);
R40 is selected from: H, Cl-Cs alkyl, or benzyl.
[~] Included in the present invention are
compounds above, wherein:
R3l is selected from the group consisting of:
WO94/22910 PCT~S94/03223
~sgQ~ --
(a) a 6 membered saturated, partially
saturated or aromatic carbocyclic ring
substituted with 0-3 R10 or R10ai
s
(b) a 8-11 membered saturated, partially
saturated, or aromatic fused bicyclic
carbocyclic ring substituted with 0-4 R10
or R10a; or
(c) a 14 membered saturated, partially
saturated, or aromatic fused tricyclic
carbocyclic ring substituted with 0-4 R10
Or Rl0a
[7] The present invention includes compounds
of formula (I) above wherein:
R31 is selected from the group consisting of:
(a) a 6 membered saturated, partially
saturated, or aromatic carbocyclic ring
of formula:
R \~
wherein any of the bonds forming the
carbocyclic ring may be a single or
double bond,
-34-
PCT~S94/03223
WO94/22910
~ 21~07~
and wherein said carbocyclic ring is
substituted independently with 0-4
R10;
(b) a 10 membered saturated, partially
saturated, or aromatic bicyclic
carbocyclic ring of formula:
~\y ~
/~\
Rl RlOa
, wherein any of the bonds forming
the carbocyclic ring may be a single
or double bond,
and wherein said carbocyclic ring is
substituted independently with 0-4
Rl0 or RlOa;
(c) a 9 membered saturated, partially
saturated, or aromatic bicyclic
carbocyclic ring of formula:
o r
Rl RlOa Rl RlOa
wherein any of the bonds forming the
carbocyclic ring may be a single or
double bond,
and wherein said carbocyclic ring is
substituted independently with 0-4
RlO or RlOa
-35-
W094/22910 PCT~S94/03223
?.~59~
[8] This invention includes compounds of
formula (I) wherein:
R31 is selected from (the dashed bond may be a
single or double bond):
\~ \~/ \~
R10 i Rl ; R10
or
Rl ' RlOa
wherein R31 may be substituted independently
with 0-3 R10 or RlOa;
n" is 0 or 1;
n' is 0-2.
[9] The present invention includes compounds of formula
(I) above wherein:
Rl and R22 are independently selected from:
phenyl, benzyl, phenethyl, phenoxy, benzyloxy,
halogen, hydroxy, nitro, cyano, Cl-c5 alkyl, C3-C6
cycloalkyl, C3-C6 cycloalkylmethyl, C7-Clo
arylalkyl, C1-Cs alkoxy, -Co2Rl3~ -C(=o)NHoR13a,
-36-
WO94122910 ~ PCT~S94/03223
~ ~15g~7~
-C(=o)NHN(Rl3)2, =NoRl3, -B(R34)(R35), C3-C6
cycloalkoxy, -oC(=o)Rl3, -C(=o)Rl3,-oc(=o)oRl3
-oRl3, -(Cl-C4 alkyl)-ORl3, -N(Rl3)2,
-oC(=o)N(Rl3)2, -NRl3C(=o)Rl3, -NRl3c(=o)oRl3
-NRl3C(=o)N(Rl3)2~ -NRl3So2N(Rl3)2, -NRl3So2Rl3
-SO3H, -So2Rl3a~ -S(=o)Rl3a~ -SRl3, -So2N(Rl3)2,
C2-C6 alkoxyalkyl, methylenedioxy, ethylenedioxy,
Cl-C4 haloalkyl, Cl-C4 haloalkoxy, Cl-C4
alkylcarbonyloxy, Cl-C4 alkylcarbonyl, Cl-C4
alkylcarbonylamino, -OCH2CO2H,
2-(l-morpholino)ethoxy, Cl-C4 alkyl (alkyl being
substituted with -N(Rl3)2, -CF3, NO2, or
-s (=O) R13a)
The present invention includes compounds
of formula (I), or a pharmaceutically
acceptable salt or prodrug form thereof
wherein:
R3l is selected from:
\[~ \~/ \[~\
Rlo ; Rlo ; Rl
~ R10~ ; or R ~ R10
wherein R3l may be substituted independently
. with 0-3 Rlo or RlOa;
-37-
W094/22910 PCT~S94/03223
~g~ ~
n" is 0 or 1;
n' is 0-2;
5 R1 and R22 are independently selected from H, C1-C4
alkyl, phenyl, benzyl, phenyl-(C2-C4)alkyl, Cl-C4
alkoxy;
R21 and R23 are independently H or C1-C4 alkyl;
R2 is H or C1-Cg alkyl;
R13 is selected independently from: H, Cl-C1o alkyl,
C3-C1o cycloalkyl, C4-C12 alkylcycloalkyl, aryl,
- (Cl-Clo alkyl)aryl, or C3-C1o alkoxyalkyl;
R13a is C1-C1o alkyl, C3-C1o cycloalkyl, C4-cl2
alkylcycloalkyl, aryl, -(C1-C1o alkyl)aryl, or
C3-C1o alkoxyalkyl;
when two R13 groups are bonded to a single N, said R13
groups may alternatively be taken together to form
- (CH2) 2-5-- or -(CH2) O (CH2)--;
25 R14 is OH, H, C1-C4 alkyl, or benzyl;
R10 and Rl0a are selected independently.from: H, C1-Cg
alkyl, phenyl, halogen, or C1-C4 alkoxy;
J is ~Ala or an L-isomer or D-isomer amino acid of
structure -N(R3)C(R4)(R5)C(=o)-, wherein:
R3 is H or CH3;
R4 is H or C1-C3 alkyl;
-38-
-
W094/22910 PCT~S94/032~
21~g~7~
R5 is H, C1-Cg alkyl, C3-C6 cycloalkyl, C3-C6
cycloalkylmethyl, C1-C6 cycloalkylethyl,
phenyl, phenylmethyl, CH2OH, CH2SH, CH20CH3,
CH2SCH3, CH2cH2scH3~ (CH2) ~3NH2~
-(CH2)~NHC(-NH)(NH2), -(CH2).~NHRl6, where s =
3-5i or
R16 is selected from:
an amine protecting group;
1-2 amino acids; or
1-2 amino acids substituted with an amine
protecting group;
R3 and R5 can alternatively be taken together to
form -(CH2)t- (t = 2-4) or -CH2SC(CH3)2-; or
R4 and R5 can alternatively be taken together to
form -(CH2)U-, where u = 2-5;
~ is an L-isomer amino acid of structure
-N(R6)CH(R7)C(=o)-, wherein:
R6 is H or C1-Cg alkyl;
R7 is
(CH2)q~NH--C~NH
~ ~NH
-- ( CH2 ) q ~ C
~ Y ~NH2, where q = O or 1;
~(CH2)rX, where r 5 3-6;
--CH2 {~--CH2X --CH2 ~ CH2X
-39-
WO94/22910 PCT~S94/03223
~9~ ~
~(CH2)mS~CH2)2X, where m = 1 or 2;
-(C3-C7 alkyl)-NH-(Cl-C6 alkyl)
-(C1-C4 alkyl) \ ~
.< NH
0-3
~(CH2)m~O~(Cl~C4 alkyl)-NH-(C1-C6 alkyl), where
m - 1 or 2;
-(CH2)m-S-(Cl-C4 alkyl)-NH-(Cl-C6 alkyl), where
m = 1 or 2; and
X is -NH2 or -NHC(=NH)(NH2); or
R6 and R7 can alternatively be taken together to
form
(CH2) nX
I
-CH2CHCH2-, where n = 0 or 1 and X is
-NH2 or -NHC(=NH)(NH2);
L is -Y(CH2)VC(=O)-, wherein:
Y is NH, O, or S; and v = 1 or 2;
~ is a D-isomer or L-isomer amino acid of structure
-NR17-CH-C(=o)
CH(R4))ql
R8 , wherein:
q' is 0-2;
-40-
WO94/22910 PCT~S94/032~
21~S07~
R17 is H, C1-C3 alkyl;
R8 is selected from:
-Co2Rl3r-so3Rl3/ -So2NHR14, -B(R34)(R35), -NHSO2CF3,
-CONHNHSO2CF3, -Po(oRl3)2/ -Po(oR13)Rl3,
-SO2NH-heteroaryl (said heteroaryl being
5-10-membered and having 1-4 heteroatoms selected
independently from N, S, or O) , -SO2NH-heteroaryl
(said heteroaryl being 5-10-membered and having 1-4
heteroatoms selected independently from N, S, or
O), -So2NHCoRl3~ -CoNHSo2Rl3a~ -CH2CoNHSo2Rl3
--NHSo2NHCoR13a, --NHCoNHSo2R13a, -So2NHCoNHR13,
-Co2Rl3b;
R13b is selected from:
(a) C1-Cg alkyl;
(b) C2-Cg alkenyl;
(c) C2-Cg alkynyl;
(d) C3-Cg cycloalkyl;
(e) Cl-Cg alkyl substituted with
(i) aryl, optionally substituted with 1-2
substituents independently selected
from halogen, phenyl, Cl-Cs alkyl,
Cl-C5 alkoxy, NO2, -s(o)o-2(cl-cs
alkyl), OH, N(R13)2, C02R13, CoN(Rl3)2
or -CVFw where v = 1 to 3 and w = 1 to
(2v+1);
(ii) C3-Cg cycloalkyl;
(iii)
(CH2)04
-41-
W094/ 910 ~ ~ PCT~S94/03223
(f) aryl, optionally substituted with 1-2
substituents independently selected from
halogen, phenyl, Cl-CS alkyl, Cl-C5 alkoxy,
NO2~ ~S(O)o-2(cl-cs alkyl), OH, N(R13)2,
C02Rl3, CoN(Rl3)2 or -CVFw where v = 1 to 3
and w = 1 to (2v+1);
(g) C2-Cg alkyl, alkenyl or alkynyl; substituted
with 1-2 substituents independently
selected from C1-C4 alkyl, C3-Cg cycloalkyl,
Cl-C5 alkoxy, phenoxy, benzyloxy, halogen,
NO2, CN, C02Rl3, CoN(Rl3)2, N(R36)CoR36~
morpholino, 2-(1-morpholino)ethoxy, N(Rl3)2,
N+(Rl3)3, OCOCH3, CF3, S(0)o-2Rl3a;
(h) CH(R36)oR38;
(i) CH(R36)oc(=o)R37;
(j) CH(R36)oc(=o)oR38;
(k) CH(R36)oC(=o)N(R37)2;
(l) CH(R36)N(R36)c(=o)R36;
(m) CH(R36)Co2R37;
(n) CH(R36)CoN(Rl3)2;
(o) CH(R36)N(Rl3)2;
(q)
,1~
O O
R39
(r)
O O
~ ~ ;
~ R39
(s)
-CH(R36)o ~;
-42-
W094/22910 PCT~S94/032~
~15~0~
.
(t)
~-~o~oo
)=(\
~,
(u)
,~30
R36 is selected independently from: H, Cl-Cg alkyl,
C3-C1o cycloalkyl, phenyl, or benzyl;
R37 is selected from:
(a) H;
(b) Cl-Cg alkyl or C3-Cg cycloalkyl, said alkyl or
cycloalkyl being substituted with 1-2 groups
independently selected from:
(i) Cl-C4 alkyl;
15 (ii) C3-Cg cycloalkyl;
(iii) C1-Cs alkoxy;
(iv) aryl substituted with 0-2 groups
independently selected from: halogen, phenyl,
Cl-C6 alkyl, Cl-C6 alkoxy, NO2, -S(Cl-Cs
alkyl), -SO(Cl-Cs alkyl), -SO2(Cl-Cs alkyl),
-OH, -N(R13)2, -Co2Rl3~ -C(=o)N(Rl3)2~ or -CVFw
where v = 1 to 3 and w = 1 to (2v+1);
(c) aryl substituted with 0-2 groups independently
selected from: halogen, phenyl, Cl-C6 alkyl,
C1-C6 alkoxy, NO2, -S(Cl-Cs alkyl), -SO(Cl-Cs
alkyl), -SO2(Cl-Cs alkyl), -OH, -N(R13)2,
-Co2Rl3~ -C(=o)N(Rl3)2~ or -CVFw where v = 1 to
3 and w = 1 to (2v+1);
R38 is selected from:
-43-
WO94/22910 PCT~S94/032
a) Cl-Cg alkyl or C3-Cg cycloalkyl, said alkyl or
cycloalkyl being substituted with 1-2 groups
independently selected from:
(i) C1-C4 alkyl;
(ii) C3-Cg cycloalkyl;
(iii) C1-Cs alkoxy;
(iv) aryl substituted with 0-2 groups
independently selected from: halogen, phenyl,
C1-C6 alkyl, C1-C6 alkoxy, NO2, -S(C1-Cs
alkyl), -SO(C1-Cs alkyl), -SO2(C1-Cs alkyl),
-OH, -N(R13)2, -Co2R13, -C(=o)N(R13)2, or -CVFw
where v = 1 to 3 and w = 1 to (2v+1);
(b) aryl substituted with 0-2 groups independently
selected from: halogen, phenyl, Cl-C6 alkyl,
Cl-C6 alkoxy, NO2, -S(Cl-Cs alkyl), -SO(Cl-Cs
alkyl), -SO2(C1-Cs alkyl), -OH, -N(R13)2,
-Co2Rl3~ -C(=o)N(R13)2, or -CVFw where v = 1 to
3 and w = 1 to (2v+1);
R39 is selected from:
(a) H
(b) C1-Cg alkyl or C3-Cg cycloalkyl, said alkyl or
cycloalkyl being substituted with 1-2 groups
independently selected from:
(i) C1-C6 alkyl;
(ii) C1-C6 alkoxy;
(iii) aryl substituted with 0-2 groups
independently selected from: halogen, phenyl,
C1-C6 alkyl, C1-C6 alkoxy, NO2, -S(C1-Cs
alkyl), -SO(C1-Cs alkyl), -SO2(C1-Cs alkyl),
-OH, -N(R13)2, -Co2Rl3~ -C(=o)N(R13)2, or -CVFw
where v = 1 to 3 and w = 1 to (2v+1);
(c) aryl substituted with 0-2 groups independently
selected from: halogen, phenyl, C1-C6 alkyl,
C1-C6 alkoxy, NO2, -S(Cl-Cs alkyl), -SO(C1-Cs
alkyl), -SO2(C1-Cs alkyl), -OH, -N(R13)2,
-44-
W094/22910 PCT~S94/03223
~ 21~907~
-Co2Rl3r -C(=o)N(Rl3)2, or -CVFw where v = 1 to
3 and w = 1 to (2v+1);
R40 is selected from: H, Cl-Cs alkyl, or benzyl.
[10] Preferred compounds of the invention are 1,3-
disubstituted phenyl compounds of the formula (II):
~ L ~ M
J/ NR2
O=C ~ CHRl
10 ~ (II)
wherein:
the phenyl ring in formula (II) may be further
substituted with 0-3 R10;
R10 is selected independently from: H, Cl-Cg alkyl,
phenyl, halogen, or C1-C4 alkoxy;
Rl is H, C1-C4 alkyl, phenyl, benzyl, or phenyl-(C1-
C4)alkyl;
R2 is H or methyl;
R13 is selected independently from: H, C1-C1o alkyl,
C3-C1o cycloalkyl, C4-C12 alkylcycloalkyl, aryl,
-(C1-C1o alkyl)aryl, or C3-C1o alkoxyalkyl;
-45-
WO94/22910 PCT~S94/03223
9~ a ~
R13a is C1-C1o alkyl, C3-C1o cycloalkyl, C4-C12
alkylcycloalkyl, aryl, -(C1-C1o alkyl)aryl, or
C3-C1o alkoxyalkyl;
when two R13 groups are bonded to a single N, said R13
groups may alternatively be taken together to form
-(CH2)2_s- or -(cH2)o(cH2)-;
R14 is OH, H, C1-C4 alkyl, or benzyl;
J is ~Ala or an L-isomer or ~-isomer amino acid of
structure -N(R3)C(R4)(R5)C(=o)-, wherein:
R3 is H or CH3;
R4 is H or C1-C3 alkyl;
R5 is H, C1-Cg alkyl, C3-C6 cycloalkyl, C3-C6
cycloalkylmethyl, C1-C6 cycloalkylethyl,
phenyl, phenylmethyl, CH2OH, CH2SH, CH2OCH3,
CH2SCH3, CH2cH2scH3~ (CH2)~NH2,
-(CH2)~NHC(=NH)(NH2), -(CH2)~NHR16, where s =
3-5; or
R16 is selected from:
an amine protecting group;
1-2 amino acids; or
1-2 amino acids substituted with an amine
protecting group;
R3 and R5 can alternatively be taken together to
form -CH2CH2CH2-; or
R4 and R5 can alternatively be taken together to
form -(CH2)U-, where u = 2-5;
-46-
WO94/22910 PCT~S94/03223
21~907~
is an L-isomer amino acid of structure
-N(R6)CH(R7)C(=o)-, wherein:
R6 is H or C1-Cg alkyl;
R7 is:
- (CH2)q ~ ~NH
NH2
~ ~NH
- (CH2)q ~ NH2, where q = 0 or 1;
~(CH2)rX, where r = 3-6;
--CH2 {~--CH2X --CH2 ~ CH2X
-(CH2)mS(CH2)2X, where m = 1 or 2;
-(C3-C7 alkyl)-NH-(Cl-C6 alkyl)
-(C1-C4 alkyl) \ ~
NH
~ Jo 3
~(CH2)m~~(C1-C4 alkyl)-NH-(Cl-C6 alkyl), where
m = 1 or ?;
~(CH2)m-S-(C1-C4 alkyl)-NH-(C1-C6 alkyl), where
m = 1 or 2; and
X is -NH2 or -NHC(=NH)(NH2), provided that X is not
~ -NH2 when r = 4; or
WO94/22910 PCT~S94/032~
9~ ~
R6 and R7 are alternatively be taken together to
form
( CH2 ) nX
-CH2CHCH2-, where n = 0,1 and X is -NH2 or
-NHC(=NH)(NH2);
L is -Y(CH2)VC(=O)-, wherein:
Y is NH, O, or S; and v = 1,2;
M is a D-isomer or L-isomer amino acid of structure
-NR17-CH-C(=o)
(CH(R4))ql
R8 , wherein:
q' is 0-2;
R17 is H, Cl-C3 alkyl;
R8 is selected from:
-Co2Rl3~-so3Rl3~ -So2NHR14, -B(R34)(R35), -NHSO2CF3,
-CONHNHSO2CF3, -PotoRl3)2~ -Po(oR13)R13,
-SO2NH-heteroaryl (said heteroaryl being
5-10-membered and having 1-4 heteroatoms selected
independently from N, S, or O) , -SO2NH-heteroaryl
(said heteroaryl being 5-10-membered and having 1-4
heteroatoms selected independently from N, S, or
O)~ -So2NHCoR13, -CoNHSo2Rl3a~ -CH2CoNHso2Rl3
-NHSo2NHCoR13a, -NHCoNHS02R13a, -So2NHCoNHR13,
-Co2Rl3b;
R13b is selected from:
(a) C2-Cg alkenyl;
-48-
WO94/22910 PCT~S94/03~3
c 7 n
(b) C2-Cg alkynyl;
(c) C1-Cg alkyl substituted with
(i) aryl, optionally substituted with 1-2
substituents independently selected
from halogen, phenyl, C1-Cs alkyl,
C1-Cs alkoxy, NO2, -s(o)o-2(cl-cs
alkyl), OH, N(R13)2, C02R13, CoN(Rl3)2
or -CVFw where v = 1 to 3 and w = 1 to
(2v+1);
(ii) C3-Cg cycloalkyl;
(iii)
~(CH2)w
A40
(e) aryl, substituted with 1-2 substituents
independently selected from halogen,
phenyl, Cl-Cs alkyl, Cl-Cs alkoxy, NO2,
-S(O)0-2(Cl-C5 alkyl), OH, N(R13)2, C02R13,
CON(R13)2 or -CVFw where v = 1 to 3 and w =
1 to (2v+1);
(f) C2-Cg alkyl, alkenyl or alkynyl; substituted
with 1-2 substituents independently
selected from C1-C4 alkyl, C3-Cg cycloalkyl,
C1-C5 alkoxy, phenoxy, benzyloxy, halogen,
NO2, CN, C02R13, CON(Rl3)2, N(R36)CoR36~
morpholino, 2-(1-morpholino)ethoxy, N(R13)2,
N+(R13)3, OCOCH3, CF3, S(0)o-2Rl3a;
(g) CH(R36)oR38;
(h) cH~R36)oc(=o)R37;
(i) CH(R36)oc(=o)oR38;
(j) CH(R36)oc(=o)N(R37)2;
(k) CH(R36)Co2R37;
(1)
-49-
W094/22910 PCT~S94/03223
9~
O O
\~1
~ R39
(m)
-CH(R36)
o
R36 is selected independently from: H, Cl-Cg alkyl,
C3-Clo cycloalkyl, phenyl, or benzyl;
R37 is selected from:
(a) H;
(b) C1-Cg alkyl or C3-Cg cycloalkyl, said alkyl or
cycloalkyl being substituted with 1-2 groups
independently selected from:
(i) Cl-C4 alkyl;
(ii) C3-Cg cycloalkyl;
(iii) Cl-Cs alkoxy;
(iv) aryl substituted with 0-2 groups
independently selected from: halogen, phenyl,
C1-C6 alkyl, C1-C6 alkoxy, NO2, -S(C1-Cs
alkyl), -SO(C1-Cs alkyl), -SO2(C1-Cs alkyl),
-OH, -N(R13)2, -Co2Rl3~ -C(=o)N(Rl3)2~ or -CVFw
where v = 1 to 3 and w = 1 to (2v+1);
(c) aryl substituted with 0-2 groups independently
selected from: halogen, phenyl, C1-C6 alkyl,
C1-C6 alkoxy, NO2, -S(C1-Cs alkyl), -SO(C1-Cs
alkyl), -SO2(Cl-Cs alkyl), -OH, -N(R13)2,
-Co2Rl3~ -C(=o)N(Rl3)2~ or -CVFw where v = 1 to
3 and w = 1 to (2v+1);
R38 is selected from:
-50-
WO94/~910 PCT~S94/032~
21S9070
(a) C1-Cg alkyl or C3-Cg cycloalkyl, said alkyl or
cycloalkyl being substituted with 1-2 groups
independently selected from:
(i) C1-C4 alkyl;
(ii) C3-Cg cycloalkyl;
(iii) Cl-Cs alkoxy;
(iv) aryl substituted with 0-2 groups
independently selected from: halogen, phenyl,
Cl-C6 alkyl, Cl-C6 alkoxy, NO2, -S(Cl-Cs
alkyl), -SO(C1-Cs alkyl), -SO2(C1-Cs alkyl),
-OH, -N(R13)2, -Co2Rl3~ -C(=o)N(Rl3)2~ or -CVFW
where v = 1 to 3 and w = 1 to (2v+1);
(b) aryl substituted with 0-2 groups independently
selected from: halogen, phenyl, Cl-C6 alkyl,
Cl-C6 alkoxy, NO2, -S(C1-Cs alkyl), -SO(C1-Cs
alkyl), -SO2(Cl-Cs alkyl), -OH, -N(R13)2,
-Co2Rl3~ -C(=o)N(R13)2, or -CVFw where v = 1 to
3 and w = l to (2v+1);
R39 is selected from:
(a) H
(b) Cl-Cg alkyl or C3-Cg cycloalkyl, said alkyl or
cycloalkyl being substituted with 1-2 groups
independently selected from:
(i) C1-C6 alkyl;
(ii) C1-C6 alkoxy;
(iii) aryl substituted with 0-2 groups
independently selected from: halogen, phenyl,
C1-C6 alkyl, Cl-C6 alkoxy, NO2, -S(Cl-Cs
alkyl), -SO(Cl-Cs alkyl), -SO2(Cl-Cs alkyl),
-OH, -N(R13)2, -Co2Rl3~ -C(=o)N(Rl3)2~ or -CVFw
where v = 1 to 3 and w = 1 to (2v+1);
(c) aryl substituted with 0-2 groups independently
~ selected from: halogen, phenyl, Cl-C6 alkyl,
Cl-C6 alkoxy, NO2, -S(C1-Cs alkyl), -SO(C1-Cs
alkyl), -SO2(C1-Cs alkyl), -OH, -N(R13)2,
WO94/22910 PCT~S94/03223
~9~ ~
-Co2Rl3~ -C(=o)N(R13)2, or -CVFw where v = 1 to
3 and w = 1 to (2v+1);
R40 is selected from: H, C1-Cs alkyl, or benzyl.
[11] Preferred compounds of the present invention are
compounds of formula (II) above, wherein:
the phenyl ring in formula (II) may be further
substituted with 0-2 R10 or R10a;
R10 or R10a are selected independently from: H, C1-C8
alkyl, phenyl, halogen, or C1-C4 alkoxy;
R1 is H;
R2 is H;
R13 is selected independently from: H, Cl-C1o alkyl,
C3-C1o cycloalkyl, C4-C12 alkylcycloalkyl, aryl,
-(Cl-Clo alkyl)aryl, or C3-Clo alkoxyalkyl;
R13a is Cl-Clo alkyl, C3-C1o cycloalkyl, C4-C12
alkylcycloalkyl, aryl, -(C1-C1o alkyl)aryl, or
C3-C1o alkoxyalkyl;
when two R13 groups are bonded to a single N, said R13
groups may alternatively be taken together to form
-tcH2)2-s- or -(cH2)o(cH2)-;
R14 is OH, H, Cl-C4 alkyl, or benzyl;
J is ~Ala or an L-isomer or D-isomer amino acid of
formula -N(R3)CH(R5)C(=o)-, wherein:
WO94122910 PCT~S94/03223
21S90~
.
R3 is H and R5 is H, CH3, CH2CH3, CH(CH3)2,
CH(CH3)CH2CH3, CH2CH2CH3, CH2CH2CH2CH3, CH2CH2SCH3,
CH2CH(CH3)2, (cH2)4NH2~ (C3-Cs alkyl)NHR16;
or
R3 is CH3 and R5 is H; or
R3 and R5 can alternatively be taken together to form
-CH2CH2CH2-;
R16 is selected from:
an amine protecting group;
1-2 amino acids;
1-2 amino acids substituted with an amine
protecting group;
is an L-isomer amino acid of formula
-N(CH3)CH(R7)C(=o)-, wherein:
R7 is -(CH2)3NHC(~NH)~NH2);
L is -NHCH2C(=O)-; and
M is a D-isomer or L-isomer amino acid of structure
-NR17-CH-C(=o)
(CH(R4))q~
R8 , wherein:
q' is l;
R4 is H or CH3;
R17 is H;
-53-
WO94122910 PCT~S94/03223
R8 is
-C02H;
-SO3H;
--C02R13b;
R13b is selected independently from:
-CH(R36)oC(=o)R37;
-CH(R36)oc(=o)oR38;
o
OJ~O
5 ~
~ ~ ~R39;
R36 is Cl-C4 linear alkyl or H;
R37 is selected from:
(a) H;
(b) C1-Cg alkyl or C3-Cg cycloalkyl, said alkyl or
cycloalkyl being substituted with 1-2 groups
independently selected from:
(i) C1-C4 alkyl;
(ii) C3-Cg cycloalkyl;
(iii) C1-Cs alkoxy;
(iv) aryl substituted with 0-2 groups
independently selected from: halogen, phenyl,
C1-C6 alkyl, C1-C6 alkoxy, NO2, -S(Cl-Cs
alkyl), -SO(C1-Cs alkyl), -SO2(C1-Cs alkyl),
-OH, -N(R13)2, -Co2Rl3t -C(=o)N(R13)2, or -CVFw
where v = 1 to 3 and w = 1 to (2v+1);
(c) aryl substituted with 0-2 groups independently
selected from: halogen, phenyl, C1-C6 alkyl,
C1-C6 alkoxy, NO2, -S(Cl-Cs alkyl), -SO(Cl-Cs
alkyl), -SO2(Cl-Cs alkyl), -OH, -N(R13)2,
WO94/22910 PCT~S94/032~
~ 21~70
-Co2Rl3~ -C(=o)N(Rl3)2r or -CVFw where v = 1 to
3 and w = 1 to (2v+1);
R38 is selected from:
(a) Cl-C8 alkyl or C3-C8 cycloalkyl, said alkyl or
cycloalkyl being substituted with 1-2 groups
independently selected from:
(i) Cl-C4 alkyl;
(ii) C3-Cg cycloalkyl;
(iii) Cl-C5 alkoxy;
(iv) aryl substituted with 0-2 groups
independently selected from: halogen, phenyl,
Cl-C6 alkyl, Cl-C6 alkoxy, NO2, -S(Cl-C5
alkyl), -SO(C1-Cs alkyl), -SO2(Cl-Cs alkyl),
-OH, -N(R13)2, -Co2Rl3~ -C(=o)N(Rl3)2~ or -CVFw
where v = 1 to 3 and w 5 1 to (2v+1);
(b) aryl substituted with 0-2 groups independently
selected from: halogen, phenyl, Cl-C6 alkyl,
Cl-C6 alkoxy, NO2, -S(Cl-Cs alkyl), -SO(Cl-Cs
alkyl), -SO2(Cl-Cs alkyl), -OH, -N(R13)2,
-Co2Rl3~ -C(=o)N(Rl3)2~ or -CVFw where v = 1 to
3 and w 5 1 to (2v+1);
R39 is C1-C4 alkyl, benzyl, or phenyl.
[12] Preferred compounds of the present invention are
compounds of formula (II), or a pharmaceutically
acceptable salt thereof, wherein:
Rl and R2 are independently selected from H, methyl;
J is selected from D-Val, D-2-aminobutyric acid, D-Leu,
D-Ala, Gly, D-Pro, D-Ser, D-Lys, ~-Ala, Pro, Phe,
~ NMeGly, D-Nle, D-Phg, D-Ile, D-Phe, D-Tyr, Ala,
N-p-azidobenzoyl-D-Lys, N-p-benzoylbenzoyl-D-Lys,
W094/229l0 PCT~S94/03223
~g~
N--tryptophanyl--D--Lys,N--o--benzylbenzoyl--D--Lys,
N-p-acetylbenzoyl-D-Lys, N-dansyl-D-Lys,
N-glycyl-D-Lys, N-glycyl-p-benzoylbenzoyl-D-Lys,
N-p-phenylbenzoyl-D-Lys, N~-m-benzoylbenzoyl-D-
Lys, N-o-benzoylbenzoyl-D-Lys;
is selected from NMeArg, Arg;
L is selected from Gly, ~-Ala, Ala;
M is selected from Asp; oMeAsp; ~MeAsp; NMeAsp; D-Asp;
Asp-(methylcarbonyloxymethyl ester);
Asp-(ethylcarbonyloxymethyl ester);
Asp-(t-butylcarbonyloxymethyl ester);
Asp-(cyclohexylcarbonyloxymethyl ester);
Asp-(1-(methylcarbonyloxy)ethyl ester);
Asp-(l-(ethylcarbonyloxy)ethyl ester);
Asp-(1-(t-butylcarbonyloxy)ethyl ester);
Asp-(1-(cyclohexylcarbonyloxy)ethyl ester);
Asp-(i-propyloxycarbonyloxymethyl ester);
Asp-(cyclohexylcarbonyloxymethyl ester);
Asp-(t-butyloxycarbonyloxymethyl ester);
Asp-(1-(i-propyloxycarbonyloxy)ethyl ester);
Asp-(1-(cyclohexyloxycarbonyloxy)ethyl ester);
Asp-(1-(t-butyloxycarbonyloxy)ethyl ester);
Asp-(dimethylaminoethyl ester);
Asp-(diethylaminoethyl ester);
Asp-((1,3-dioxa-5-methyl-cyclopenten-2-one-4-
yl)methyl ester);
Asp-((5-(t-butyl)-1,3-dioxa-cyclopenten-2-one-4-
yl)methyl ester);
Asp-((1,3-dioxa-5-phenyl-cyclopenten-2-one-4-
yl)methyl ester);
Asp-(1-(2-(2-methoxypropyl)carbonyloxy)ethyl
ester).
-5~-
WO94/22910 PCT~S94/032~
~ g O 7 ~
[13] Preferred compounds of the present invention are
compounds of formula (II), or a pharmaceutically
acceptable salt thereof, wherein:
R1 and R2 are independently selected from H, methyl;
J is selected from: D-Val, D-2-aminobutyric acid, D-Leu,
D-Ala, Gly, D-Pro, D-Ser, D-Lys, ~-Ala, Pro, Phe,
NMeGly, D-Nle, D-Phg, D-Ile, D-Phe, D-Tyr, Ala;
~ is selected from NMeArg;
L is Gly;
M is selected from Asp; aMeAsp; ~MeAsp; NMeAsp; D-Asp;
Asp-(methylcarbonyloxymethyl ester);
Asp-(ethylcarbonyloxymethyl ester);
Asp-(t-butylcarbonyloxymethyl ester);
Asp-(cyclohexylcarbonyloxymethyl ester);
Asp-(1-(methylcarbonyloxy)ethyl ester);
Asp-(1-(ethylcarbonyloxy)ethyl ester);
Asp-(1-(t-butylcarbonyloxy)ethyl ester);
Asp-(1-(cyclohexylcarbonyloxy)ethyl ester);
Asp-(i-propyloxycarbonyloxymethyl ester);
Asp-(cyclohexylcarbonyloxymethyl ester);
Asp-(t-butyloxycarbonyloxymethyl ester);
Asp-(1-(i-propyloxycarbonyloxy)ethyl ester);
Asp-(1-(cyclohexyloxycarbonyloxy)ethyl ester);
Asp-(1-(t-butyloxycarbonyloxy)ethyl ester);
Asp-(dimethylaminoethyl ester);
Asp-(diethylaminoethyl ester);
- Asp-((1,3-dioxa-5-methyl-cyclopenten-2-one-4-
yl)methyl ester);
-57-
W094/22910 ~CT~S94/032~
~5~orl ~ - --
Asp-((5-(t-butyl)-1,3-dioxa-cyclopenten-2-one-4-
yl)methyl ester);
Asp-((1,3-dioxa-5-phenyl-cyclopenten-2-one-4-
yl)methyl ester);
Asp-(1-(2-(2-methoxypropyl)carbonyloxy)ethyl
ester).
[14] Specifically preferred compounds of the present
invention are the following compounds and
pharmaceutically acceptable salts thereof:
The compound of formula (II) wherein Rl and R2 are H; J
is D-Val; K is NMeArg; L is Gly; and M is
Asp-(methylcarbonyloxymethyl ester).
The compound of formula (II) wherein R1 and R2 are H; J
is D-Val; K is NMeArg; L is Gly; and M is
Asp-(ethylcarbonyloxymethyl ester).
The compound of formula (II) wherein R1 and R2 are H; J
is D-Val; K is NMeArg; L is Gly; and M is Asp-(t-
butylcarbonyloxymethyl ester).
The compound of formula (II) wherein R1 and R2 are H; J
is D-Val; K is NMeArg; L is Gly; and M is
Asp-(cyclohexylcarbonyloxymethyl ester).
.
The compound of formula (II) wherein R1 and R2 are H; J
is D-Val; K is NMeArg; L is Gly; and M is Asp-(1-
(methylcarbonyloxy)ethyl ester).
The compound of formula (II) wherein R1 and R2 are H; J
is D-Val; K is NMeArg; L is Gly; and M is Asp-(1-
(ethylcarbonyloxy)ethyl ester).
-58-
WO94122910 PCT~S94/032~
2159070
The compound of formula (II) wherein Rl and R2 are H; J
is D-Val; K is NMeArg; L is Gly; and M is Asp-(l-(t-
butylcarbonyloxy)ethyl ester).
-
The compound of formula (II) wherein Rl and R2 are H; Jis D-Val; K is NMeArg; L is Gly; and M is Asp-(l-
(cyclohexylcarbonyloxy)ethyl ester).
The compound of formula (II) wherein Rl and R2 are H; J
is D-Val; K is NMeArg; L is Gly; and M is Asp-(i-
propyloxycarbonyloxymethyl ester).
The compound of formula (II) wherein Rl and R2 are H; J
is D-Val; K is NMeArg; L is Gly; and M is
Asp-(cyclohexylcarbonyloxymethyl ester).
The compound of formula (II) wherein Rl and R2 are H; J
is D-Val; K is NMeArg; L is Gly; and M is Asp-(t-
butyloxycarbonyloxymethyl ester).
The compound of formula (II) wherein Rl and R2 are H; J
is D-Val; K is NMeArg; L is Gly; and M is Asp-(l-(i-
propyloxycarbonyloxy)ethyl ester).
The compound of formula (II) wherein Rl and R2 are H; J
is D-Val; K is NMeArg; L is Gly; and M is Asp-(l-
(cyclohexyloxycarbonyloxy)ethyl ester).
The compound of formula (II) wherein Rl and R2 are H; J
is D-Val; K is NMeArg; L is Gly; and M is Asp-(l-(t-
butyloxycarbonyloxy)ethyl ester).
The compound of formula (II) wherein Rl and R2 are H; J
is D-Val; K is NMeArg; L is Gly; and M is
Asp-(dimethylaminoethyl ester).
-59-
W094/~910 PCT~S94/032~
2~S9~
The compound of formula (II) wherein R1 and R2 are H; J
is D-Val; K is NMeArg; L is Gly; and M is
Asp-(diethylaminoethyl ester).
The compound of formula (II) wherein R1 and R2 are H; J
is D-Val; K is NMeArg; L is Gly; and M is Asp-((1,3-
dioxa-5-methyl-cyclopenten-2-one-4-yl)methyl ester).
The compound of formula (II) wherein Rl and R2 are H; J
is D-Val; K is NMeArg; L is Gly; and M is Asp-((5-(t-
butyl)-1,3-dioxa-cyclopenten-2-one-4-yl)methyl ester).
The compound of formula (II) wherein R1 and R2 are H; J
is D-Val; K is NMeArg; L is Gly; and M is Asp-((1,3-
dioxa-5-phenyl-cyclopenten-2-one-4-yl)methyl ester).
The compound of formula (II) wherein R1 and R2 are H; J
is D-Val; K is NMeArg; L is Gly; and M is Asp-(1-(2-(2-
methoxypropyl)carbonyloxy)ethyl ester).
The compound of formula (II) wherein R1 and R2 are H; Jis D-2-aminobutyric acid; K is NMeArg; L is Gly; and M
is Asp-(methylcarbonyloxymethyl ester).
The compound of formula (II) wherein R1 and R2 are H; J
is D-2-aminobutyric acid; K is NMeArg; L is Gly; and M
is Asp-(ethylcarbonyloxymethyl ester).
The compound of formula (II) wherein R1 and R2 are H; J
is D-2-aminobutyric acid; K is NMeArg; L is Gly; and M
is Asp-(t-butylcarbonyloxymethyl ester).
-60-
WO94122910 PCT~S94/032~
21~g~7~
.
The compound of formula (II) wherein Rl and R2 are H; J
is D-2-aminobutyric acid; K is NMeArg; L is Gly; and M
is Asp-(cyclohexylcarbonyloxymethyl ester).
The compound of formula (II) wherein Rl and R2 are H; J
is D-2-aminobutyric acid; K is NMeArg; L is Gly; and M
is Asp-(l-(methylcarbonyloxy)ethyl ester).
The compound of formula (II) wherein Rl and R2 are H; J
is D-2-aminobutyric acid; K is NMeArg; L is Gly; and M
is Asp-~l-(ethylcarbonyloxy)ethyl ester).
The compound of formula (II) wherein Rl and R2 are H; J
is D-2-aminobutyric acid; K is NMeArg; L is Gly; and M
is Asp-(l-(t-butylcarbonyloxy)ethyl ester).
The compound of formula (II) wherein Rl and R2 are H; J
is D-2-aminobutyric acid; K is NMeArg; L is Gly; and M
is Asp-(l-(cyclohexylcarbonyloxy)ethyl ester).
The compound of formula (II) wherein Rl and R2 are H; J
is D-2-aminobutyric acid; K is NMeArg; L is Gly; and M
is Asp-(i-propyloxycarbonyloxymethyl ester).
The compound of formula (II) wherein Rl and R2 are H; J
is D-2-aminobutyric acid; K is NMeArg; L is Gly; and M
is Asp-(cyclohexylcarbonyloxymethyl ester).
The compound of formula (II) wherein Rl and R2 are H; J
is D-2-aminobutyric acid; K is NMeArg; L is Gly; and M
is Asp-(t-butyloxycarbonyloxymethyl ester).
-
The compound of formula (II) wherein Rl and R2 are H; Jis D-2-aminobutyric acid; K is NMeArg; L is Gly; and M
is Asp-(l-(i-propyloxycarbonyloxy)ethyl ester).
--~1--
WO94/22910 PCT~S94/032~
O~ Q - : ~
The compound of formula (II) wherein R1 and R2 are H; J
is D-2-aminobutyric acid; K is NMeArg; L is Gly; and M
is Asp-(1-(cyclohexyloxycarbonyloxy)ethyl ester).
The compound of formula (II) wherein R1 and R2 are H; J
is D-2-aminobutyric acid; K is NMeArg; L is Gly; and M
is Asp-(1-(t-butyloxycarbonyloxy)ethyl ester).
The compound of formula (II) wherein R1 and R2 are H; J
is D-2-aminobutyric acid; K is NMeArg; L is Gly; and M
is Asp-(dimethylaminoethyl ester).
The compound of formula (II) wherein R1 and R2 are H; J
is D-2-aminobutyric acid; K is NMeArg; L is Gly; and M
is Asp-(diethylaminoethyl ester).
The compound of formula (II) wherein R1 and R2 are H; J
is D-2-aminobutyric acid; K is NMeArg; L is Gly; and M
is Asp-((1,3-dioxa-5-methyl-cyclopenten-2-one-4-
yl)methyl ester).
The compound of formula (II) wherein R1 and R2 are H; J
is D-2-aminobutyric acid; K is NMeArg; L is Gly; and M
is Asp-((5-(t-butyl)-1,3-dioxa-cyclopenten-2-one-4-
yl)methyl ester).
The compound of formula (II) wherein R1 and R2 are H; J
is D-2-aminobutyric acid; K is NMeArg; L is Gly; and M
is Asp-((1,3-dioxa-5-phenyl-cyclopenten-2-one-4-
yl)methyl ester).
The compound of formula (II) wherein R1 and R2 are H; J
is D-2-aminobutyric acid; K is NMeArg; L is Gly; and M
is Asp-(1-(2-(2-methoxypropyl)carbonyloxy)ethyl ester).
-62-
W094/22910 PCT~S94/032~
21~gO70
In the present invention it has been discovered
that the compounds above are useful as inhibitors of
glycoprotein IIb/IIIa (GPIIb/IIIa). As discussed above,
GPIIb/IIIa mediates the process of platelet activation
and aggregation. The compounds of the present invention
inhibit the activation and aggregation of platelets
induced by all known endogenous platelet agonists.
The present invention also provides methods for the
treatment (including prevention) of conditions involving
platelet activation and aggregation, such as arterial or
venous cardiovascular or cerebrovascular thromboembolic
disorders, including, for example, thromboembolic
disorders associated with unstable angina, first or
recurrent myocardial infarction, ischemic sudden death,
transient ischemic attack, stroke, atherosclerosis, deep
vein thrombosis, pulmonary embolism, or diabetes, by
administering to a host in need of such treatment a
pharmaceutically effective amount of the compounds
described above. The compounds of the present invention
are useful for inhibiting the binding of fibrinogen to
blood platelets, inhibiting aggregation of blood
platelets, treating thrombus formation or embolus
formation, or preventing thrombus or embolus formation
in a mammal. The compounds of the invention may be used
as a medicament for blocking fibrinogen from acting at
its receptor site in a mammal.
The compounds of the present invention can also be
combined or co-administered with suitable anti-coagulant
or coagulation inhibitory agents, such as heparin or
warfarin, or anti-platelet or platelet inhibitory
- agents, such as aspirin, piroxicam or ticlopidine.
Further, the compounds of this invention may be combined
- or co-administered with thrombin inhibitors such as
boropeptides, hirudin or argatroban. The compounds of
the present invention may also be combined or
-63-
WO94/22910 PCT~S94/032
co-administered with thrombolytic or fibrinolytic
agents, such as plasminogen activators, anistreplase,
urokinase, or streptokinase. The compounds of the
present invention may also be combined or
co-administered with combinations of the foregoing
agents and/or with other therapeutic agents. Such
combination products may be employed to achieve
synergistic effects or effects additive to those
provided by the compounds of the present invention, such
as, for example, in such uses as described above,
particularly in the treatment, including prevention, of
thromboembolic disorders.
The GPIIb/IIIa antagonists of the present invention
inhibit platelet aggregation at the final common pathway
required for platelet aggregation induced by any of the
known platelet activators or even their combinations.
On the other hand, platelet granular secretions, of
various important biomolecules from the a-granule tPAI-
l) or the dense granule (serotonin) are not affected bythe GPIIb/IIIa antagonist. These molecules secreted
from platelets might play an important role in arterial
vasospasm (serotonin) and in reducing the efficiency of
the natural lytics (PAI-l). Hence, the combination of
the compounds of the present invention with other drugs
which may affect these mechanisms and may thereby
provide a particularly effective therapy for many
different heterogenous thromboembolic disorders.
The GPIIb/IIIa antagonists of the present invention
with high affinity for the platelet GPIIb/IIIa receptor
(Kd < O.0l ~M) are expected to be very effective not
only in preventing thrombosis formation, but also in
accelerating lysis of platelet rich thrombi, thereby
providing a greater utility of such antiplatelet agents
in the acute and chronic thromboembolic disorders. Such
-64-
WO94/22910 215 9 ~ 7 ~ PCT~S94/032~
a strategy may be an effective adjunct therapy with
thrombolytic therapy. Indeed, platelet activation after
thrombolytic therapy may have a significant role in the
delay of reperfusion and abrupt closure (reocclusion).
The term anti-coagulant agents (or coagulation
inhibitory agents), as used herein, denotes agents that
inhibit blood coagulation. Such agents include
warfarin, heparin, or low molecular weight heparin
(LMWH), including pharmaceutically acceptable salts or
prodrugs thereof. For reasons of efficacy, the
preferable anti-coagulant agents are warfarin or heparin
or LMWH. The warfarin employed herein, may be, for
example, crystalline warfarin or amorphous sodium
warfarin. The heparin employed herein may be, for
example, the sodium or sulfate salts thereof.
The term anti-platelet agents ~or platelet
inhibitory agents), as used herein, denotes agents that
inhibit platelet function such as by inhibiting the
aggregation, adhesion or granular secretion of
platelets. Such agents include the various known
non-steroidal anti-inflammatory drugs (NSAIDS) such as
aspirin, ibuprofen, naproxen, sulindac, indomethacin,
mefenamate, droxicam, diclofenac, sulfinpyrazone, and
piroxicam, including pharmaceutically acceptable salts
or prodrugs thereof. Of the NSAIDS, aspirin
(acetylsalicyclic acid or ASA), which has been well
researched and widely used with good results, and
piroxicam, which exerts its anti-platelet effect when
dosed once daily, are preferred compounds, especially
- aspirin. Piroxicam is commercially available from
Pfizer Inc. (New York, NY), as FELDANE . Other suitable
anti-platelet agents include ticlopidine, including
pharmaceutically acceptable salts or prodrugs thereof.
Ticlopidine is also a preferred compound since it is
-65-
W094122910 PCT~S94/03223
known to be gentle on the gastro-intestinal tract in
use. Still other suitable platelet inhibitory agents
include thromboxane-A2-receptor antagonists and
thromboxane-A2-synthetase inhibitors, as well as
pharmaceutically acceptable salts or prodrugs thereof.
-66-
W094/22910 PCT~S94/032~
~ 215~07~
The phrase thrombin inhibitors ~or anti-thrombin
agents), as used herein, denotes inhibitors of the
serine protease thrombin. By inhibiting thrombin,
various thrombinmediated processes, such as
thrombin-mediated platelet activation (that is, for
example, the aggregation of platelets, and/or the
granular secretion of plasminogen activator inhibitor-l
and/or serotonin) and/or fibrin formation are disrupted.
Such inhibitors include boropeptides, hirudin and
argatroban, including pharmaceutically acceptable salts
and prodrugs thereof. Preferably the thrombin
inhibitors are boropeptides. By boropeptides, it is
meant, N-acetyl and peptide derivatives of boronic acid,
such as C-terminal a-aminoboronic acid derivatives of
lysine, ornithine, arginine, homoarginine and
corresponding isothiouronium analogs thereof. The term
hirudin, as used herein, includes suitable derivatives
or analogs of hirudin, referred to herein as hirulogs,
such as disulfatohirudin.
Preferable boropeptide thrombin inhibitors include
compounds described in Kettner et al., U.S. Patent No.
5,187,157 and European Patent Application Publication
Number 293 881 A2, the disclosures of which are hereby
incorporated herein by reference. Other suitable
boropeptide thrombin inhibitors include those disclosed
in PCT Patent Application Publication Number 92/07869
and European Patent Application Publication Number 471
651 A2, the disclosures of which are hereby incorporated
herein by reference, in their entirety.
The phrase thrombolytics (or fibrinolytic) agents
~ ~or thrombolytics or fibrinolytics), as used herein,
denotes agents that lyse blood clots (thrombi). Such
agents include tissue plasminogen activator,
anistreplase, urokinase or streptokinase, including
pharmaceutically acceptable salts or prodrugs thereof.
-67-
W094/229l0 PCT~S941032~
~9~
Tissue plasminogen activator (tPA) is commercially
available from Genentech Inc., South San Francisco,
California. The term anistreplasè, as used herein,
refers to anisoylated plasminogen streptokinase
activator complex, as described, for example, in
European Patent Application No. 0 28 489, the
disclosures of which are hereby incorporated herein by
reference herein, in their entirety. Anistreplase is
commercially available from the Beecham Group,
Middlesex, England, under the trademark EMINASETM. The
term urokinase, as used herein, is intended to denote
both dual and single chain urokinase, the latter also
being referred to herein as prourokinase.
Combination products, where the cyclic
compounds of the invention are combined or
co-administered with suitable anti-coagulant agents,
antiplatelet agents, thrombin inhibitors, and/or
thrombolytic agents, may afford an efficacy advantage
over the compounds and agents alone, and may do so while
permitting the use of lower doses of each. A lower
dosage minimizes the potential of side effects, thereby
providing an increased margin of safety.
Clinical studies using anti-coagulant agents alone,
including crystalline sodium warfarin, have provided
evidence of their efficacy in the treatment or secondary
prevention of coronary artery disease. Of three
published, randomized, controlled trials of the
treatment of acute myocardial infarction, oral anti-
coagulants significantly reduced overall mortality and
the frequency of reinfarction in one study. Of the four
published large, randomized, controlled trials of oral
anti-coagulants in the secondary prevention of
myocardial infarction, three suggested a reduction in
the incidence of reinfarction and early mortality. One
additional study, the Warfarin Reinfarction Study, has
also recently demonstrated a significant reduction in
-68-
WO94/22910 PCT~S94/032~
2159~7~
.
mortality, reinfarction, and stroke in people with a
previous myocardial infarction who were treated with
warfarin as compared to those treated with placebo.
The results of studies utilizing anti-platelet
agents such as acetylsalicylic acid (ASA) alone in the
prevention and treatment of coronary artery disease have
also been promising. In patients with unstable angina,
ASA has been demonstrated to reduce the incidence of
subsequent myocardial infarction and mortality in two
large, randomized, double-blind, placebo-controlled
clinical studies. In addition, ASA has been approved
for use in the secondary prevention of myocardial
infarction, based on data from several trials which,
when pooled, suggested a reduction in reinfarction and
mortality. Furthermore, two recent studies evaluating
ASA in the primary prevention of coronary artery disease
have reported either a dramatic or inconsequential
benefit. In addition to their utility in coronary
artery disease, agents that inhibit platelet function
such as ASA and ticlopidine have been shown to be
effective in the prevention of stroke in people with
cerebrovascular disease. Pooled data from nine
randomized trials have provided overwhelming evidence of
the efficacy of ASA alone in reducing the risk of
completed stroke in people with transient ischemic
attacks (TIAs). Recently, ticlopidine alone has also
been demonstrated to have efficacy in treating TIAs.
With regard to thrombin inhibitors, such as
boropeptides, studies have demonstrated that such
compounds provide excellent candidates for the control
of thrombinmediated processes. Studies with hirudin,
~ another thrombin inhibitor, have shown this agent to be
an effective compound in the treatment of venous and
~ arterial thrombosis.
Current therapy in the treatment of patients with
acute myocardial infarction includes thrombolytics such
-69-
W094/22910 PCT~S94/03223
2~9~
as plasminogen activators such as tPA, streptokinase, or
urokinase. These standard thrombolytics, when employed
alone, promote the generation of plasmin, which degrades
platelet-rich fibrin clots.
Thromboembolic disorders are known, however, to
have a diverse pathophysiological makeup. There is a
need for a therapeutic approach to the treatment of
these disorders which takes into account the diverse
pathophysiological makeup of such diseases, and which
includes components ameliorating each of the various
pathophysiological aspects. A combination product
containing an anti-coagulant agent such as warfarin or
heparin, or an antiplatelet agent such as aspirin,
piroxicam or ticlopidine, or a thrombin inhibitor such
as a boropeptide, hirudin or argatroban, or a
thrombolytic agent such as tissue plasminogen activator,
anistreplase, urokinase or streptokinase, or
combinations thereof, in combination with a novel cyclic
compound of the invention, can provide such an approach.
In addition, by administering lower doses of each, which
is feasible where an additive or synergistic effect is
involved, the incidence of any side effects associated
with each alone at higher doses may be significantly
reduced. Also, where a convenient single dosage form is
offered, as in a preferred embodiment of the invention,
it is generally accepted that such increased convenience
to the patient results in an increase in compliance.
Also, a single dosage form would reduce the likelihood
of patient confusion often associated with concurrent
dosing of medication not available in a single dosage
form. The present combinations of an anticoagulant
agent and a compound of this invention, or an
anti-platelet agent and a compound of this invention, or
a thrombin inhibitor and a compound of this invention,
or a thrombolytic agent and a compound of this
-70-
WO94/22910 PCT~S94/03~
21~907~
.
invention, or combinations thereof, are directed to
meeting these, as well as other, needs.
GPIIb/IIIa is known to be overexpressed in
metastatic tumor cells. The compounds or combination
products of the present invention may also be useful for
the treatment, including prevention, of metastatic
cancer.
The compounds herein described may have asymmetric
centers. Unless otherwise indicated, all chiral,
diastereomeric and racemic forms are included in the
present invention. Many geometric isomers of olefins,
C=N double bonds, and the like can also be present in
the compounds described herein, and all such stable
isomers are contemplated in the present invention. It
will be appreciated that compounds of the present
invention contain asymmetrically substituted carbon
atoms, and may be isolated in optically active or
racemic forms. It is well known in the art how to
prepare optically active forms, such as by resolution of
racemic forms or by synthesis, from optically active
starting materials. Two distinct isomers (cis and
trans) of the peptide bond are known to occur; both can
also be present in the compounds described herein, and
all such stable isomers are contemplated in the present
invention. Unless otherwise specifically noted, the L-
isomer of the amino acid is used at positions J, K, L,
and M of the compounds of the present invention. Except
as provided in the preceding sentence, all chiral,
diastereomeric, racemic forms and all geometric isomeric
forms of a structure are intended, unless the specific
- stereochemistry or isomer form is specifically
indicated. The D and L-isomers of a particular amino
acid are designated herein using the conventional 3-
letter abbreviation of the amino acid, as indicated by
the following examples: D-Leu, D-Leu, L-Leu, or L-Leu.
WO94/22910 PCT~S94103223
When any variable (for example, Rl through R8, m,
n, p, X, Y, etc.) occurs more than one time in any
constituent or in any formula, its definition on each
occurrence is independent of its definition at every
other occurrence. Thus, for example, if a group is
shown to be substituted with 0-2 Rl1, then said group
may optionally be substituted with up to two Rll and R
at each occurrence is selected independently from the
defined list of possible Rll. Also, by way of example,
for the group -N(R13)2, each of the two R13 substituents
on N is independently selected from the defined list of
possible R13.
When a bond to a substituent is shown to cross the
bond connecting two atoms in a ring, then such
substituent may be bonded to any atom on the ring.
Combinations of substituents and/or variables are
permissible only if such combinations result in stable
compounds.
By "stable compound" or "stable structure" is meant
herein a compound that is sufficiently robust to survive
isolation to a useful degree of purity from a reaction
mixture, and formulation into an efficacious therapeutic
agent.
The term "substituted", as used herein, means that
an one or more hydrogen on the designated atom is
replaced with a selection from the indicated group,
provided that the designated atom's normal valency is
not exceeded, and that the substitution results in a
stable compound. When a substitent is keto (i.e., =O),
then 2 hydrogens on the atom are replaced.
As used herein, "alkyl" is intended to include both
branched and straight-chain saturated aliphatic
hydrocarbon groups having the specified number of carbon
atoms; "haloalkyl" is intended to include both branched
and straight-chain saturated aliphatic hydrocarbon
WO94122910 PCT~S94/032~
21~gQ70
groups having the specified number of carbon atoms,
substituted with 1 or more halogen (for example -cvFw
where v = 1 to 3 and w = 1 to (2v+1)); "alkoxy"
represents an alkyl group of indicated number of carbon
atoms attached through an oxygen bridge; "cycloalkyl" is
intended to include saturated ring groups, including
mono-,bi- or poly-cyclic ring systems, such as
cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl,
cycloheptyl, cyclooctyl and adamantyl; and "biycloalkyl"
is intended to include saturated bicyclic ring groups
such as [3.3.0]bicyclooctane, [4.3.0]bicyclononane,
[4.4.0]bicyclodecane (decalin), [2.2.2]bicyclooctane,
and so forth. "Alkenyl" is intended to include
hydrocarbon chains of either a straight or branched
configuration and one or more unsaturated carbon-carbon
bonds which may occur in any stable point along the
chain, such as ethenyl, propenyl and the like; and
"alkynyl" is intended to include hydrocarbon chains of
either a straight or branched configuration and one or
more triple carbon-carbon bonds which may occur in any
stable point along the chain, such as ethynyl, propynyl
and the like.
The phrase "boronic acid" as used herein means a
group of the formula -B(R34)(R35), wherein R34 and R35
are independently selected from: -OH; -F; -NR13R14; or
C1-Cg-alkoxy; or R34 and R35 can alternatively be taken
together to form: a cyclic boron ester where said chain
or ring contains from 2 to 20 carbon atoms and,
optionally, 1-4 heteroatoms independently selected from
N, S, or O; a divalent cyclic boron amide where said
- chain or ring contains from 2 to 20 carbon atoms and,
optionally, 1-4 heteroatoms independently selected from
- N, S, or O; a cyclic boron amide-ester where said chain
or ring contains from 2 to 20 carbon atoms and,
optionally, 1-4 heteroatoms independently selected from
WO94122910 PCT~S94/032~
2~39~ ~
N, S, or O. Such cyclic boron esters, boron amides, or
boron amide-esters may also be optionally substituted
with 1-5 groups independently selected from Rll.
Boron esters include boronic acid protecting
groups, including moieties derived from diols, for
example pinanediol and pinacol to form pinanediol
boronic acid ester and the pinacol boronic acid,
respectively. Other illustrations of diols useful for
deriving boronic acid esters are perfluoropinacol,
ethylene glycol, diethylene glycol, 1,2-ethanediol,
1,3-propanediol, 1,2-propanediol, 1,2-butanediol,
1,4-butanediol, 2,3-butanediol, 2,3-hexanediol,
1,2-hexanediol, catechol, 1,2-diisopropylethanediol,
5,6-decanediol, 1,2-dicyclohexylethanediol.
"Halo" or "halogen" as used herein refers to
fluoro, chloro, bromo and iodo; and "counterion" is used
to represent a small, negatively charged species such as
chloride, bromide, hydroxide, acetate, sulfate and the
like.
As used herein, "aryl" or "aromatic residue" is
intended to mean phenyl or naphthyl. As used herein,
"carbocycle" or "carbocyclic residue" is intended to
mean any stable 3- to 7- membered monocyclic or bicyclic
or 7- to 14-membered bicyclic or tricyclic or an up to
26-membered polycyclic carbon ring, any of which may be
saturated, partially unsaturated, or aromatic. Examples
of such carbocyles include, but are not limited to,
cyclopropyl, cyclopentyl, cyclohexyl, phenyl, biphenyl,
naphthyl, indanyl, adamantyl, or tetrahydronaphthyl
~tetralin).
As used herein, the term "heterocycle" or
"heterocyclic ring system" is intended to mean a stable
5- to 7- membered monocyclic or bicyclic or 7- to 10-
membered bicyclic heterocyclic ring which may besaturated, partially unsaturated, or aromatic, and which
WO94/22910 PCT~S94/032~
~ 21~9~7~
consists of carbon atoms and from 1 to 4 heteroatoms
selected independently from the group consisting of N, O
and S and wherein the nitrogen and sulfur heteroatoms
may optionally be oxidized, and the nitrogen may
optionally be quaternized, and including any bicyclic
group in which any of the above-defined heterocyclic
rings is fused to a benzene ring. The heterocyclic ring
may be attached to its pendant group at any heteroatom
or carbon atom which results in a stable structure. The
heterocyclic rings described herein may be substituted
on carbon or on a nitrogen atom if the resulting
compound is stable. Examples of such heterocycles
include, but are not limited to, benzopyranyl,
thiadiazine, tetrazolyl, benzofuranyl, benzothiophenyl,
indolene, quinoline, isoquinolinyl or benzimidazolyl,
piperidinyl, 4-piperidone, 2-pyrrolidone,
tetrahydrofuran, tetrahydroquinoline,
tetrahydroisoquinoline, decahydroquinoline,
octahydroisoquinoline, azocine, triazine (including
1,2,3-, 1,2,4-, and 1,3,5-triazine), 6H-1,2,5-
thiadiazine, 2H,6H-1,5,2-dithiazine, thiophene,
tetrahydrothiophene, thianthrene, furan, pyran,
isobenzofuran, chromene, xanthene, phenoxathiin,
2H-pyrrole, pyrrole, imidazole, pyrazole, thiazole,
isothiazole, oxazole (including 1,2,4- and 1,3,4-
oxazole), isoxazole, triazole, pyridine, pyrazine,
pyrimidine, pyridazine, indolizine, isoindole, 3H-
indole, indole, lH-indazole, purine, 4H-quinolizine,
isoquinoline, quinoline, phthalazine, naphthyridine,
quinoxaline, quinazoline, cinnoline, pteridine,
4aH-carbazole, carbazole, B-carboline, phenanthridine,
- acridine, perimidine, phenanthroline, phenazine,
phenarsazine, phenothiazine, furazan, phenoxazine,
isochroman, chroman, pyrrolidine, pyrroline,
imidazolidine, imidazoline, pyrazolidine, pyrazoline,
piperazine, indoline, isoindoline, quinuclidine, or
-75-
WO94122910 PCT~S94/03223
~9~
morpholine. Also included are fused ring and spiro
compounds containing, for example, the above
heterocycles.
As used herein, the term "any group that, when
administered to a mammalian subject, cleaves to form a
free hydroxyl, amino or sulfhydryl" means any group
bonded to an O, N, or S atom, respectively, which is
cleaved from the O, N, or S atom when the compound is
administered to a mammalian subject to provide a
compound having a remaining free hydroxyl, amino, or
sulfhydryl group, respectively. Examples of groups
that, when administered to a mammalian subject, are
cleaved to form a free hydroxyl, amino or sulfhydryl,
include but are not limited to, C1-C6 alkyl substituted
with 0-3 R11, C3-C6 alkoxyalkyl substituted with 0-3
R11, C1-C6 alkylcarbonyl substituted with 0-3 R11, C1-C6
alkoxycarbonyl substituted with 0-3 R11, C1-C6
alkylaminocarbonyl substituted with 0-3 R11, benzoyl
substituted with 0-3 R12, phenoxycarbonyl substituted
with 0-3 R12, phenylaminocarbonyl substituted with 0-3
R12. Examples of groups that, when administered to a
mammalian subject, are cleaved to form a free hydroxyl,
amino or sulfhydryl, include hydroxy, amine or
sulfhydryl protecting groups, respectively.
As used herein, the term "amine protecting group"
means any group known in the art of organic synthesis
for the protection of amine groups. Such amine
protecting groups include those listed in Greene,
"Protective Groups in Organic Synthesis" John Wiley &
Sons, New York (1981) and "The Peptides: Analysis,
Sythesis, Biology, Vol. 3, Academic Press, New York
(1981), the disclosure of which is hereby incorporated
by reference. Any amine protecting group known in the
-76-
WO94122910 PCT~S94/032~
215907~
art can be used. Examples of amine protecting groups
include, but are not limited to, the following: l) acyl
types such as formyl, trifluoroacetyl, phthalyl, and
p-toluenesulfonyl; 2) aromatic carbamate types such as
benzyloxycarbonyl (Cbz) and substituted
benzyloxycarbonyls, l-(p-biphenyl)-l-
methylethoxycarbonyl, and 9-fluorenylmethyloxycarbonyl
(Fmoc); 3) aliphatic carbamate types such as tert-
butyloxycarbonyl (Boc), ethoxycarbonyl,
diisopropylmethoxycarbonyl, and allyloxycarbonyl; 4)
cyclic alkyl carbamate types such as
cyclopentyloxycarbonyl and adamantyloxycarbonyl; 5)
alkyl types such as triphenylmethyl and benzyl; 6)
trialkylsilane such as trimethylsilane; and 7) thiol
containing types such as phenylthiocarbonyl and
dithiasuccinoyl. Also included in the term 1'amine
protecting group" are acyl groups such as azidobenzoyl,
p-benzoylbenzoyl, o-benzylbenzoyl, p-acetylbenzoyl,
dansyl, glycyl-p-benzoylbenzoyl, phenylbenzoyl,
m-benzoylbenzoyl, benzoylbenzoyl.
As used herein, "pharmaceutically acceptable salts"
refer to derivatives of the disclosed compounds wherein
the parent compound of formula (I) is modified by making
acid or base salts of the compound of formula (I).
Examples of pharmaceutically acceptable salts include,
but are not limited to, mineral or organic acid salts of
basic residues such as amines; alkali or organic salts
of acidic residues such as carboxylic acids; and the
like.
"Prodrugs" are considered to be any covalently
bonded carriers which release the active parent drug
- according to formula (I) in vivo when such prodrug is
administered to a mammalian subject. Prodrugs of the
compounds of formula (I) are prepared by modifying
functional groups present in the compounds in such a way
that the modifications are cleaved, either in routine
W094/22910 PCT~S94/032~
~9~ a
manipulation or in vivo, to the parent compounds.
Prodrugs include compounds of formula (I) wherein
hydroxy, amine, or sulfhydryl groups are bonded to any
group that, when administered to a mammalian subject,
cleaves to form a free hydroxyl, amino, or sulfhydryl
group, respectively. Examples of prodrugs include, but
are not limited to, acetate, formate and benzoate
derivatives of alcohol and amine functional groups in
the compounds of formula (I); and the like.
Pharmaceutically acceptable salts of the compounds
of the invention can be prepared by reacting the free
acid or base forms of these compounds with a
stoichiometric amount of the appropriate base or acid in
water or in an organic solvent, or in a mixture of the
two; generally, nonaqueous media like ether, ethyl
acetate, ethanol, isopropanol, or acetonitrile are
preferred. Lists of suitable salts are found in
R~m;naton's PhArm~ceuti~l Sciences, 17th ed., Mack
Publishing Company, Easton, PA, 1985, p. 1418, the
disclosure of which is hereby incorporated by reference.
The term "amino acid" as used herein means an
organic compound containing both a basic amino group and
an acidic carboxyl group. Included within this term are
modified and unusual amino acids, such as those
disclosed in, for example, Roberts and Vellaccio ~1983)
The Peptides, 5: 342-429, the teaching of which is
hereby incorporated by reference. Modified or unusual
amino acids which can be used to practice the invention
include, but are not limited to, D-amino acids,
hydroxylysine, 4-hydroxyproline, ornithine,
2,4-diaminobutyric acid, homoarginine, norleucine,
N-methylaminobutyric acid, naphthylalanine,
phenylglycine, ~-phenylproline, tert-leucine,
4-aminocyclohexylalanine, N-methyl-norleucine,
3,4-dehydroproline, 4-aminopiperidine-4-carboxylic acid,
WO94/22910 PCT~S94/03223
2I ~90 7~
.
6-aminocaproic acid, trans-4-(aminomethyl)-
cyclohexanecarboxylic acid, 2-, 3-, and 4-(aminomethyl)-
benzoic acid, 1-aminocyclopentanecarboxylic acid,
l-aminocyclopropanecarboxylic acid, and 2-benzyl-5-
aminopentanoic acid.
The term "amino acid residue" as used herein means
that portion of an amino acid (as defined herein) that
is present in a peptide.
The term "peptide" as used herein means a linear
compound that consists of two or more amino acids (as
defined herein) that are linked by means of a peptide
bond. The term "peptide" also includes compounds
containing both peptide and non-peptide components, such
as pseudopeptide or peptide mimetic residues or other
non-amino acid components. Such a compound containing
both peptide and non-peptide components may also be
referred to as a "peptide analog".
A "pseudopeptide" or "peptide mimetic" is a
compound which mimics the structure of an amino acid
residue or a peptide, for example, by using linking
groups other than amide linkages between the peptide
mimetic and an amino acid residue (pseudopeptide bonds)
and/or by using non-amino acid substituents and/or a
modified amino acid residue.
A "pseudopeptide residue" means that portion of an
pseudopeptide or peptide mimetic ~as defined herein)
that is present in a peptide.
The term "peptide bond" means a covalent amide
linkage formed by loss of a molecule of water between
the carboxyl group of one amino acid and the amino group
of a second amino acid.
- The term "pseudopeptide bonds" includes peptide
bond isosteres which may be used in place of or as
substitutes for the normal amide linkage. These
substitute or amide "equivalent" linkages are formed
from combinations of atoms not normally found in
-79-
W094l22910 PCT~S94/03223
peptides or proteins which mimic the spatial
requirements of the amide bond and which should
stabilize the molecule to enzymatic degradation.
The compounds of the present invention can be
prepared in a number of ways well known to one skilled
in the art of organic synthesis. Preferred methods
include but are not limited to those methods described
below.
The following abbreviations are used herein:
D-Abu D-2-aminobutyric acid
~-Ala, b-Ala or
~Ala 3-aminopropionic acid
Boc t-butyloxycarbonyl
Boc-iodo-Mamb t-butyloxycarbonyl-3-aminomethyl-4-iodo-
benzoic acid
Boc-Mamb t-butyloxycarbonyl-3-aminomethylbenzoic
acid
Boc-ON [2-(tert-butyloxycarbonyloxylimino)-2-
phenylacetonitrile
C12Bzl dichlorobenzyl
CBZ or Cbz Carbobenzyloxy
25 DCC dicyclohexylcarbodiimide
DIEA diisopropylethylamine
di-NMeOrn N-aMe-N-~Me-ornithine
DMAP 4-dimethylaminopyridine
HBTU 2-(lH-Benzotriazol-1-yl)-1,1,3,3-
tetramethyluronium hexafluorophosphate
NMeArg or
MeArg a-N-methyl arginine
NMeAmf N-Methylaminomethylphenylalanine
NMeAsp a-N-methyl aspartic acid
35 NMeGly or
MeGly N-methyl glycine
-80-
W094t22910 PCT~S94/03223
2159078
.
NMe-Mamb N-methyl-3-aminomethylbenzoic acid
NMM N-methylmorpholine
OcHex O-cyclohexyl
OBzl O-benzyl
TBTU 2-(lH-Benzotriazol-1-yl)-1,1,3,3-
tetramethyluronium tetrafluoroborate
Tos tosyl
The following conventional three-letter amino acid
abbreviations are used herein; the conventional one-
letter amino acid abbreviations are not used herein:
Ala = alanine
Arg = arginine
15 Asn = asparagine
Asp = aspartic acid
Cys - cysteine
Gln = glutamine
Glu = glutamic acid
20 Gly = glycine
His = histidine
Ile ~ isoleucine
Leu = leucine
Lys - lysine
25 Met = methionine
Nle - norleucine
Phe = phenylalanine
Phg = phenylglycine
Pro = proline
30 Ser = serine
Thr = threonine
- Trp = tryptophan
Tyr = tyrosine
- Val = valine
Pept;de Synthes;s
-81-
r~
WO94/22910 PCT~S94/032~
z~90~
The compounds of the present invention can be
synthesized using standard synthetic methods known to
those skilled in the art. Preferred methods include but
are not limited to those methods described below.
Generally, peptides are elongated by deprotecting
the a-amine of the C-terminal residue and coupling the
next suitably protected amino acid through a peptide
linkage using the methods described. This deprotection
and coupling procedure is repeated until the desired
sequence is obtained. This coupling can be performed
with the constituent amino acids in a stepwise fashion,
or condensation of fragments (two to several amino
acids), or combination of both processes, or by solid
phase peptide synthesis according to the method
originally described by Merrifield, J. Am. Chem. Soc.,
85, 2149-2154 (1963), the disclosure of which is hereby
incorporated by reference.
The compounds of the invention may also be synthesized
using automated peptide synthesizing equipment. In
addition to the foregoing, procedures for peptide
synthesis are described in Stewart and Young, "Solid
Phase Peptide Synthesis", 2nd ed, Pierce Chemical Co.,
Rockford, IL (1984); Gross, Meienhofer, Udenfriend,
Eds., "The Peptides: Analysis, Synthesis, Biology, Vol.
1, 2, 3, 5, and 9, Academic Press, New York, (1980-
1987); Bodanszky, "Peptide Chemistry: A Practical
Textbook", Springer-Verlag, New York (1988)i and
Bodanszky et al. "The Practice of Peptide Sythesis"
Sprin~er-Verlag, New York (1984), the disclosures of
which are hereby incorporated by reference.
The coupling between two amino acid derivatives, an
amino acid and a peptide, two peptide fragments, or the
cyclization of a peptide can be carried out using
standard coupling procedures such as the azide method,
mixed carbonic acid anhydride (isobutyl chloroformate)
-82-
WO94/22910 PCT~S94/032~
215907~
.
method, carbodiimide (dicyclohexylcarbodiimide,
diisopropylcarbodiimide, or water-soluble carbodiimides)
method, active ester (p-nitrophenyl ester, N-
hydroxysuccinic imido ester) method, Woodward reagent K
method, carbonyldiimidazole method, phosphorus reagents
such as BOP-Cl, or oxidation-reduction method. Some of
these methods (especially the carbodiimide) can be
enhanced by the addition of l-hydroxybenzotriazole.
These coupling reactions may be performed in either
solution (liquid phase) or solid phase.
The functional groups of the constituent amino
acids must be protected during the coupling reactions to
avoid undesired bonds being formed. The protecting
groups that can be used are listed in Greene,
"Protective Groups in Organic Synthesis" John Wiley &
Sons, New York (1981) and "The Peptides: Analysis,
Sythesis, Biology, Vol. 3, Academic Press, New York
(1981), the disclosure of which is hereby incorporated
by reference.
The a-carboxyl group of the C-terminal residue is
usually protected by an ester that can be cleaved to
give the carboxylic acid. These protecting groups
include: 1) alkyl esters such as methyl and t-butyl, 2)
aryl esters such as benzyl and substituted benzyl, or 3)
esters which can be cleaved by mild base treatment or
mild reductive means such as trichloroethyl and phenacyl
esters. In the solid phase case, the C-terminal amino
acid is attached to an insoluble carrier (usually
polystyrene). These insoluble carriers contain a group
which will react with the carboxyl group to form a bond
which is stable to the elongation conditions but readily
~ cleaved later. Examples of which are: oxime resin
(DeGrado and Kaiser (1980) J. Org. Chem. 45, 1295-1300)
~ chloro or bromomethyl resin, hydroxymethyl resin, and
aminomethyl resin. Many of these resins are
-83-
WO94/229~0~7 ~ PCT~S94/032~
commercially available with the desired C-terminal amino
acid already incorporated.
The a-amino group of each amino acid must be
protected. Any protecting group known in the art can be
used. Examples of these are: 1) acyl types such as
formyl, trifluoroacetyl, phthalyl, and p-
toluenesulfonyl; 2) aromatic carbamate types such as
benzyloxycarbonyl ~Cbz) and substituted
benzyloxycarbonyls, 1-(p-biphenyl)-1-
methylethoxycarbonyl, and 9-fluorenylmethyloxycarbonyl
(Fmoc); 3) aliphatic carbamate types such as tert-
butyloxycarbonyl (Boc), ethoxycarbonyl,
diisopropylmethoxycarbonyl, and allyloxycarbonyl; 4)
cyclic alkyl carbamate types such as
cyclopentyloxycarbonyl and adamantyloxycarbonyl; 5)
alkyl types such as triphenylmethyl and benzyl; 6)
trialkylsilane such as trimethylsilane; and 7) thiol
containing types such as phenylthiocarbonyl and
dithiasuccinoyl. The preferred a-amino protecting group
is either Boc or Fmoc. Many amino acid derivatives
suitably protected for peptide synthesis are
commercially available.
The a-aminO protecting group is cleaved prior to
the coupling of the next amino acid. When the Boc group
is used, the methods of choice are trifluoroacetic acid,
neat or in dichloromethane, or HCl in dioxane. The
resulting ammonium salt is then neutralized either prior
to the coupling or in situ with basic solutions such as
aqueous buffers, or tertiary amines in dichloromethane
or dimethylformamide. When the Fmoc group is used, the
reagents of choice are piperidine or substituted
piperidines in dimethylformamide, but any secondary
amine or aqueous basic solutions can be used. The
deprotection is carried out at a temperature between 0
C and room temperature.
-84-
WO94/22910 PCT~S94/03223
21~50~
Any of the amino acids bearing side chain
functionalities must be protected during the preparation
of the peptide using any of the above-identified groups.
Those skilled in the art will appreciate that the
selection and use of appropriate protecting groups for
these side chain functionalities will depend upon the
amino acid and presence of other protecting groups in
the peptide. The selection of such a protecting group
is important in that it must not be removed during the
deprotection and coupling of the a-amino group.
For example, when Boc is chosen for the a-amine
protection the following protecting groups are
acceptable: p-toluenesulfonyl (tosyl) moieties and nitro
for arginine; benzyloxycarbonyl, substituted
benzyloxycarbonyls, or tosyl for lysine; benzyl or
alkyl esters such as cyclopentyl for glutamic and
aspartic acids; benzyl ethers for serine and threonine;
benzyl ethers, substituted benzyl ethers or 2-
bromobenzyloxycarbonyl for tyrosine; p-methylbenzyl, p-
methoxybenzyl, acetamidomethyl, benzyl, or t-
butylsulfonyl for cysteine; and the indole of tryptophan
can either be left unprotected or protected with a
formyl group.
When Fmoc is chosen for the a-amine protection
usually tert-butyl based protecting groups are
acceptable. For instance, Boc can be used for lysine,
tert-butyl ether for serine, threonine and tyrosine, and
tert-butyl ester for glutamic and aspartic acids.
Once the elongation and cyclization of the peptide
is completed all of the protecting groups are removed.
For the liquid phase synthesis the protecting groups are
~ removed in whatever manner as dictated by the choice of
protecting groups. These procedures are well known to
those skilled in the art.
When a solid phase synthesis is used, the peptide
should be removed from the resin without simultaneously
-85-
W094/22~ ~ PCT~S94/032
removing protecting groups from functional groups that
might interfere with the cyclization process. Thus, if
the peptide is to be cyclized in solution, the cleavage
conditions need to be chosen such that a free a-
carboxylate and a free a-amino group are generated
without simultaneously removing other protecting groups.
Alternatively, the peptide may be removed from the resin
by hydrazinolysis, and then coupled by the azide method.
Another very convenient method involves the synthesis of
peptides on an oxime resin, followed by intramolecular
nucleophilic displacement from the resin, which
generates a cyclic peptide (Osapay, Profit, and Taylor
(1990) Tetrahedron Letters 43, 6121-6124). When the
oxime resin is employed, the Boc protection scheme is
generally chosen. Then, the preferred method for
removing side chain protecting groups generally involves
treatment with anhydrous HF containing additives such as
dimethyl sulfide, anisole, thioanisole, or p-cresol at 0
C. The cleavage of the peptide can also be
accomplished by other acid reagents such as
trifluoromethanesulfonic acid/trifluoroacetic acid
mixtures.
Unusual amino acids used in this invention can be
synthesized by standard methods familiar to those
skilled in the art ("The Peptides: Analysis, Sythesis,
Biology, Vol. 5, pp. 342-449, Academic Press, New York
(1981)). N-Alkyl amino acids can be prepared using
procedures described in previously (Cheung et al.,
(1977) Can. J. Chem. 55, 906; Freidinger et al., (1982)
J. Org. Chem. 48, 77 (1982)), which are incorporated
here by reference.
The compounds of the present invention may be
prepared using the procedures further detailed below as
well as the procedures described in PCT Patent
Application International Publication Number
-86-
WO94/22910 21~ 9 ~ ~ ~ PCT~S94/032~
WO 93/07170, the disclosure of which is hereby
incorporated herein by reference. Those compounds
referred to by Example Number which are not detailed
herein are disclosed in detail in PCT International
Publication Number WO 93/07170.
Representative materials and methods that may be
used in preparing the compounds of the invention are
described further below.
Manual solid phase peptide synthesis was performed
in 25 mL polypropylene filtration tubes purchased from
BioRad Inc. Oxime resin (substitution level = 0.96
mmol/g) was prepared according to published procedures
(DeGrado and Kaiser (1980) J. Org. Chem. 45, 1295). All
chemicals and solvents (reagent grade) were used as
supplied from the vendors cited without further
purification. t-Butyloxycarbonyl (Boc) amino acids and
other starting amino acids may be obtained commercially
from Bachem Inc., Bachem Biosciences Inc. (Philadelphia,
PA), Advanced ChemTech (Louisville, KY), Peninsula
Laboratories (Belmont, CA), or Sigma (St. Louis, MO).
2-(lH-Benzotriazol-1-yl)-1,1,3,3-tetramethyluronium
hexafluorophosphate (HBTU) and TBTU were purchased from
Advanced ChemTech. N-methylmorpholine ~NMM), m-cresol,
D-2-aminobutyric acid (Abu), trimethylacetylchloride,
diisopropylethylamine (DIEA), 3-cyanobenzoic acid and
[2-(tert-butyloxycarbonyloxylimino)-phenylacetonitrile]
(Boc-ON) were purchased from Aldrich Chemical Company.
Dimethylformamide (DMF), ethyl acetate, chloroform
(CHCl3), methanol (MeOH), pyridine and hydrochloric acid
(HCl) were obtained from Baker. Acetonitrile,
dichloromethane (DCM), acetic acid (HOAc),
- trifluoroacetic acid (TFA), ethyl ether, triethylamine,
acetone, and magnesium sulfate were purchased from EM
Science. Palladium on carbon catalyst (10% Pd) was
purchased from Fluka Chemical Company. Absolute ethanol
was obtained from Quantum Chemical Corporation. Thin
-87-
WO94/22910 ~ ~ PCT~S94/032
layer chromatography (TLC~ was performed on Silica Gel
60 F2s4 TLC plates (layer thickness 0.2 mm) which were
purchased from EM Separations. TLC visualization was
accomplished using UV light, iodine, and/or ninhydrin
spray. Melting points were determined using a Thomas
Hoover or Electrothermal 9200 melting point apparatus
and are uncorrected. HPLC analyses were performed on
either a Hewlett Packard 1090, Waters Delta Prep 3000,
Rainin, or DuPont 8800 system. NMR spectra were
recorded on a 300 MHz General Electric QE-300, Varian
300, or Varian 400 spectrometer. Fast atom bombardment
mass spectrometry (FAB-MS) was performed on a VG Zab-E
double-focusing mass spectrometer using a Xenon FAB gun
as the ion source or a Finnigan MAT 8230.
Synthesis of 3 ~n~ 4-sllhstitute~ Roc-~m; nomethylhenzoic
Ac;d Der;v~tives
3 and 4-substituted Boc-aminomethylbenzoic acid
derivatives useful as intermediates in the synthesis of
the compounds of the invention are prepared using
standard procedures, for example, as described in ~ett.
Lett., 4393 (1975); Modern Synthetic Reactions, H.O.
House (1972); or Harting et al. ~. Am. Chem. Soc., 50:
3370 (1928), and as shown schematically below.
O OH O ~ OH o OH
,~CN ~d ~ NH~ HCI BoC~ON ,X Nl I BOC
EtOH-HCI ~J Et, N, pH = 9
3-Am;nomethylhenzoic Ac;~-HCl
3-Cyanobenzoic acid (10.0 g, 68 mmol) was dissolved in
200 ml ethanol by heating in a 35-50 C water bath.
-88-
WO94/22910 21 S 9 0 7 ~ PCT~S94/032~
-
Concentrated HCl (6.12 ml, 73 mmol) was added and the
solution was transferred to a 500 ml nltrogen-flushed
round bottom flask containing palladium on carbon
catalyst (1.05 g, 10% Pd/C). The suspension was stirred
under an atmosphere of hydrogen for 38 hours, filtered
through a scintered glass funnel, and washed thoroughly
with H2O. The ethanol was removed under reduced
pressure and the remaining aqueous layer, which
contained a white solid, was diluted to 250 ml with
additional H2O. Ethyl ether (250 ml) was added and the
suspension was transferred to a separatory funnel. Upon
vigorous shaking, all solids dissolved and the aqueous
layer was then washed two times with ether, evaporated
under reduced pressure to a volume of 150 ml, and
lyophilized to give the title compound (3-
aminomethylbenzoic acid HCl) (8.10 g, 64%) as a beige
solid. lH NMR (D20) 4.27 (s, 2H), 7.60 (t, lH), 7.72
(d,lH), 8.06 (d, 2H).
t-Rutyloxyc~rhonyl-3-~m;nomethylhenzoic Acid (Boc-~mh)
The title compound was prepared according to a
modification of standard procedures previously reported
in the literature (Itoh, Hagiwara, and Kamiya (1975)
Tett. Lett., 4393). 3-Aminomethylbenzoic acid
(hydrochloride salt) (3.0 g, 16.0 mmol) was dissolved in
60 ml H2O. To this was added a solution of Boc-ON (4.33
g, 17.6 mmol) in 60 ml acetone followed by
triethylamine (5.56 ml, 39.9 mmol). The solution
turned yellow and the pH was adjusted to 9 (wet pH
paper) by adding an additional 1.0 ml (7.2 mmol)
triethylamine. The solution was stirred overnight at
room temperature at which time the acetone was removed
under reduced pressure and the remaining aqueous layer
was washed three times with ether. The aqueous layer
was then acidified to pH 2 with 2N HCl and then
-89-
WO94/22910 PCT~S94/032~
?,~5~rtQ
extracted three times with ethyl acetate. The combined
organic layers were washed three times with H20, dried
over anhydrous magnesium sulfate, and evaporated to
dryness under reduced pressure. The material was
recrystallized from ethyl acetate/ hexane to give two
crops of the title compound (2.58 g, 64%) as an off-
white solid. mp 123-125C ;1H NMR (CDC13) 1.47 (s, 9
H), 4.38 (br s, 2 H), 4.95 (br s, lH), 7.45 (t, lH),
7.55 (d, lH), 8.02 (d, 2H).
t-Butyloxycaxbonyl-N-methyl-3-aminome~hylbenzoic Acid
(Boc-NMeM~mh)
The title compound can be prepared according to
standard procedures, for examples, as disclosed in
Olsen, J. Org. Chem. (1970) 35: 1912), and as shown
schematically below.
O
BOC - NH ~ OH ~ . I ~ OMe
DMF, 45O
1) NaOH/EtOH, BOC N ~ OH
2) H Me
Synthesis of Aminome~ylhenzoic Aci~ lo~s
Intermediates of the formula below may be prepared
using standard synthetic procedures, for example, as
shown in the indicated reaction schemes shown below.
-90-
WO94/22910 PCT~S94/03223
215g~
o NH-BOC
HO~R
For R = CH3, CH2CH3, CH2CH2CH3, CH2CH2CH2CH3,
CH(CH3)2, C(CH3)3, CH(CH3)CH2CH3, benzyl, cyclopentyl,
cyclohexyl; see Scheme 1.
For R = CH3, CH2CH2CH2CH3, phenyl; see Scheme 2.
For R = CH3, phenyl; see Scheme 3 and ~.
-91-
WO 94122910 PCTIUS94/03223
~59orl~
Scheme 1:
o N 0 10% Pd-C/H2/ o NH2.HCI
HoJ~3f DBU/CH311THF MeOJ~N HcvEtoH J~J
HN=~ CH2C12
Ph
Ph
Ph ~F'h
O NH-BOC (i) C~ . h O N~Ph O N
HOJ~ (ii) BOC-ON J~ Bl RX M~oJ~3J
.~ch~me 2:
N H29O4 aq/A NaN(SiMe3)2 N.SiM~3
~ HOJ~ ~ NaO~J
(i) RLi
(ii) H20/HCI
O NH-BOC BOC-ON NH2.HCI
HO~R ~ HOJ~R
--92--
WO94/~910 PCT~S94/032~
21~9~
Scheme 3:
O NH2HCI
HO~RNH4oAclNa(cN)BH3 ,~`R
CH30H/Mol. Sieves l~l
Boc-On/Acetone
O NH-BOC
HOJ~R
Scheme 4:
O O
HO~R (i) NH20H.HCI/EtOH/Pyr. J~,J~
(ii) Pd-C/EtOH/HClcon./H2 ~
Boc-On/Acetona
o NH-BOC
HO J~R
3- Ul-(t-hutyloxycarbonyl)aminolethylbenzoic ~cid
(ROC-Me~MR)
The title compound for the purpose of this
invention was prepared according to the Scheme 4
(above).
3-Acetylbenzoic acid (0.50 g, 3 mmol),
hydroxylamine hydrochloride (0.70 g, 10 mmol) and
pyridine (0.70 ml, 9 mmol) were refluxed in 10 ml
ethanol, for 2 h. Reaction mixture was concentrated,
residue triturated with water, filtered and dried. Oxime
was isolated as a white solid (0.51 g ; 94.4% yield).
-93-
WO94/22910 PCT~S94/032
S~
?~9~1 ~
HNMR (CD30D) 7.45-8.30(m, 4H), 2.30(s, 3H). MS (CH4-CI)
[M+H-O] = 164.
A solution of the oxime (0.5`1 g, 3 mmol) in
ethanol, containing 10% Pd on carbon (1.5 g) and conc.
HCl (0.25 ml, 3 mmol) was hydrogenated at 30 psi H2
pressure in a Parr hydrogenator for 5 h. Catalyst was
filtered and the filtrate concentrated. Residue was
triturated with ether. Amine hydrochloride was isolated
as a white solid (0.48 g ; 85.7% yield). 1HNMR (CD30D)
7.6-8.15(m, 4H), 4.55(q, lH), 1.70(s, 3H). MS [M+H] =
166.
Amine hydrochloride (0.40 g, 2 mmol) was dissolved
in 15 ml water. A solution of BOC-ON (0.52 g, 2.1 mmol)
in 15 ml acetone was added, followed by the addition of
triethylamine (0.8 ml, 6 mmol). Reaction was allowed to
proceed for 20 h. Reaction mixture was concentrated,
partitioned between ethyl acetate and water. Aqueous
layer was acidified to pH 2 using 10% HCl solution.
Product was extracted in ethyl acetate, which after the
usual work up and recrystallization from ethyl
acetate/hexane, gave the title compound as a white solid
(0.30 g ; 57% yield). m.p. 116-118 C.
1HNMR (CDCl3) 7.35-8.2(m, 4H), 4.6(bs, 1.5H), 1.50(d,
3H), 1.40(s, 9H). MS (NH3-CI) [M+NH4] = 283.
3-rl'-(t-hutyloxyc~rho~yl)~m; nolhenzylhenzoic ~c;A
(ROC-PhM~R)
The title compound for the purpose of this
invention was prepared according to the Scheme 4
(above), by the procedure similar to that for the methyl
derivative.
A solution of 3-benzoylbenzoic acid (2.00 g, 9
mmol), hydroxylamine hydrochloride (2.00 g, 29 mmol) and
pyridine (2.00 ml, 25 mmol) in ethanol was refluxed for
12 h. After the usual extractive work up, white solid
-94-
WO94122910 PCT~S94/032~
21~9~7~
was obtained (2.41 g). The product still contained
traces of pyridine, but was used in the next step
without further purification.
The crude product (2.00 g, ~8 mmol) was dissolved
in 200 ml ethanol. 10% Pd-C (2.00 g) and con. HCl (1.3
ml, 16 mmol) were added. Reaction mixture was
hydrogenated at 30 psi for 1 h. The catalyst was
filtered and the reaction mixture concentrated. Upon
trituration of the residue with ether and drying under
vacuum, amine hydrochloride was obtained as a white
solid (2.12 g ; 97% yield). 1HNMR (CD30D) 7.4-8.15(m,
10H), 5.75(s, lH). MS (CH4-CI) [M+H-OH] = 211.
Amine hydrochloride (1.00 g, 4 mmol) was converted
to its BOC-derivative by a procedure similar to the
methyl case. 0.60 g (48% yield) of the recrystallized
(from ethanol/hexane) title compound was obtained as a
white solid. m.p. 190-192 C. 1HNMR (CD30D) 7.2-8.0(m,
10H), 5.90 (2s, lH, 2 isomers), 1.40(s, 9H). MS (NH3-CI)
[M~NH4-C4Hg] = 289
t-Rutyloxyc~r~onyl-D-2-~mlnohutyric Acid
The title compound was prepared by a modification
of procedures previously reported in the literature
(Itoh, Hagiwara, and Kamiya (1975) Tett. Lett., 4393),
as shown in the scheme below.
~ O
NH2~OH 9 Et N ' + OJl`NH
O O
D-2-amlnobutyrlc acid
D-2-aminobutyric acid (1.0 g, 9.70 mmol) was
dissolved in 20 ml H2O and a solution of Boc-ON (2.62 g,
10.6 mmol) in 20 ml acetone was added. A white
precipitate formed which dissolved upon addition of
-95-
WO941~910 PCT~S94/032~
triethylamine (3.37 ml, 24.2 mmol) to give a pale yellow
solution (pH = 9, wet pH paper). The solution was
stirred at room temperature overnight at which time the
acetone was removed under reduced pressure. The
remaining aqueous layer was extracted with ether three
times, acidified to pH 2 with concentrated HCl, and then
extracted with ethyl acetate three times. The combined
organic layers were dried over anhydrous magnesium
sulfate and evaporated under reduced pressure to give t-
butyloxycarbonyl-D-2-aminobutyric acid as an oil (2.05
g,greater than quantitative yield, contains solvent),
which was used without further purification. lH NMR
(CDC13) 0.98 (t, 3H), 1.45 (s, 9H), 1.73 (m, lH), 1.90
(m, lH), 4.29 (m, lH), 5.05 (m, lH).
Synthes;s of t-Rutyloxyc~rhonyl-3-~m;noDhenyl~cetic Acid
t-Butyloxycarbonyl-3-aminophenylacetic acids useful
as intermediates in the synthesis of the compounds of
the invention are prepared using standard procedures,
for example, as described in Collman and Groh (1982) J.
Am. C~em. Soc., 104: 1391, and as shown schematically
below.
~C02H ~CO2H ~CO2H
NH~OH ~ t-Bu02COCO2t-Elu
Feso~-7H2o I DIEA
25 No2 H,N~H H' `CO2t-Bu
t-Rutyloxyc~rbonyl-3-~m;nophenylacetic Acid
A solution of 3-aminophenylacetic acid ~Aldrich, 10
g, 66 mmol), di-tert-butyl dicarbonate (15.8 g, 72
mmol), and DIEA (8.6 g, 66 mmol) in 50 ml of
dichloromethane was stirred overnight at room
-96-
WO94122910 ~1 ~ g 0 7 ~ PCT~S94/032~
.
temperature. The reaction mixture was concentrated,
partitioned between dichloromethane-H2O, the water layer
was separated, acidified to pH 3 with lN HCl, and
extracted with dichloromethane. The extracts were washed
with H2O, brine, dried over anhydrous sodium sulfate,
and evaporated to dryness under reduced pressure. This
material was purified by recrystallization from heptane
to provide the title compound (3.7 g, 22%) as a white
solid. mp 105C; lH NMR (CDC13) 7.35 (s, lH), 7.25 (m,
3H), 6.95 (m, lH), 6.60 (br s, lH), 3.65 (s, 2H), 1.50
(s, 9H).
Synthes;s of 4 5 And 6-SI7hstitute~ 3-
Am;nomethylhenzoic Aci~-~Cl ~n~ 4 5 And 6-Sllhst;tuted
t-Rutyloxyc~rbonyl-3-Am;nomethylhenzo;c Ac;~ Deriv~t;ves
4, 5, and 6-Substituted 3-aminomethylbenzoic
acid-HCl, and 4, 5, and 6-substituted t-
butyloxycarbonyl-3-aminomethylbenzoic acid derivatives
useful as intermediates in the synthesis of the
compounds of the invention are prepared using standard
procedures, for example, as described in Felder et al
Helv. Chim. Acta, 48: 259 (1965); de Diesbach Helv.
Chim. Acta, 23: 1232 (1949); Truitt and Creagn J. Org.
25 Chem., 27: 1066 (1962); or Sekiya et al Chem. Pharm.
Bull., 11: 551 (1963), and as shown schematically below.
-97-
WO94/22910 PCT~S94/03~
~' C02H Cl2CHCONHCH20H CI~N CO2H
Z~XX conc H~SO~ Cl H~
HCI
HOAc
H20
Coc-HN ~c02H 80c-ON, Et~U HCI-H2N ~CO2H
Y Y
Synthesis of 9-Chloro-3-~m;nomethylhenzoic Acid-HC1
The title compound was prepared by modification of
procedures previously reported in the literature (Felder
et al ~1965) ~elv. Chim. Acta, 48: 259). To a solution
of 4-chlorobenzoic acid (15.7 g, 100 mmol) in 150 ml of
concentrated sulfuric acid was added N-hydroxymethyl
dichloroacetamide (23.7 g, 150 mmol) in portions. The
reaction mixture was stirred at room temperature for 2
days, poured onto 375 g of ice, stirred for 1 hour, the
solid was collected by filtration, and washed with H20.
The moist solid was dissolved in 5% sodium bicarbonate
solution, filtered, and acidified to pH 1 with
concentrated HCl. The solid was collected by filtration,
washed with H20, and air-dryed overnight to give 4-
chloro-3-dichloroacetylaminomethylbenzoic acid (26.2 g,
89%) as a white powder.
A suspension of 4-chloro-3-
dichloroacetylaminomethylbenzoic acid (26.2 g, 88 mmol)
in 45 ml of acetic acid, 150 ml of concentrated HCl, and
-98-
W094/22910 PCT~S94/032~
21~gO7~
150 ml of H2O was heated to reflux for 3 hours, filtered
while hot, and allowed to cool to room temperature. The
solid was collected by filtration, washed with ether,
washed with acetone-ether, and air-dryed overnight to
give the title compound (7.6 g, 39%) as off-white
crystals. mp 278-9C; lH NMR (D6-DMSO) 13.40 (br s, lH),
8.75 (br s, 3H), 8.20 (s, lH), 7.95 (dd, lH), 7.70 (d,
lH), 4.20 (br s, 2H).
t-Rutyloxyc~rhonyl-4-chloro-3-~m;nomethylhenzo;c Ac;A
A suspension of 4-chloro-3-aminomethylbenzoic
acid-HCl (6.7 g, 30 mmol) and triethylamine (9.3 g, 92
mmol) in 50 ml of H2O, was added to a solution of Boc-ON
(9.2 g, 38 mmol) in 50 ml of tetrahydrofuran cooled to
0C. The reaction mixture was stirred at room
temperature overnight, and the volatile compounds were
removed by concentration under reduced pressure. The
residue was diluted with H2O, washed with ether,
acidified to pH 3 with lN HCl, and extracted with ethyl
acetate. The extracts were washed with H2O, brine, dried
over anhydrous magnesium sulfate, and evaporated to
dryness under reduced pressure. This material was
triturated with ether-hexane to provide the title
compound (7.4 g, 87%) as a white powder. mp 159C (dec);
H NMR (D6-DMSO) 13.20 ~br s, lH), 7.90 (s, lH), 7.80
(dd, lH), 7.60 (br s, lH), 7.55 (d, lH), 4.20 (br d,
2H), 1.40 (s, 9H).
4 ~n~ 6-.~l~hst;tllte~ t-RutyloxycArhonyl-3-
~m; nomethylhenzoic Aci~ Der;v~tives
.
The compounds listed below were prepared using the
general procedure described above for t-
butyloxycarbonyl-4-chloro-3-aminomethylbenzoic acid.
_99_
WO94t22910 PCT~S94/032~
~"~s~la ~
Boc-HN ~C02H
R10 ~ Rl~
Rl0a ~10 mp C
H Cl 159
H I 168
H Me 155
H MeO 171
Cl H 150
I H 182
- Me H 166
MeO H 79
Synthes;s of 2-Am; nomethylhenzoic Ac; d-HCl And 2-
A~;nomethyl~henyl A cetic Ac;~-~Cl
2-Aminomethylbenzoic acid HCl and 2-
aminomethylphenylacetic acid-HCl useful as intermediates
in the synthesis of the compounds of the invention are
prepared using standard procedures, for example, as
described in Naito et al J. Antibiotics, 30 : 698 11977);
or Young and Sweet ~. Am. Chem. Soc., 80: 800 (1958),
and as shown schematically below.
~ TMSN3,
\2SO4 CO2H
~ ~ HCI, ~ )n
1. NBS ~ ~NH ~NH2-HCI
~C02H ~. NH40H n s 0,1
~Me
--100--
WO94122910 2 I ~ 9 0 ~ ~ PCT~Sg4/032~
2-Am;nomethylDhenyl~cetic Ac;~ ~-T~ct~m
The title compound was prepared by modification of
procedures previously reported in the literature (Naito
et al. (1977) J. Antibiotics, 30: 698). To an ice-cooled
suspension of 2-indanone (10.8 g, 82 mmol) and
azidotrimethylsilane (9.4 g, 82 mmol) in 115 ml of
chloroform was added 25 ml of concentrated sulfuric acid
at a rate to maintain the temperature between 30-40C.
After an additional 3 hours, the reaction mixture was
poured onto ice, and the water layer was made basic with
concentrated ammonium hydroxide. The chloroform layer
was separated, washed with H2O, brine, dried over
anhydrous magnesium sulfate, and evaporated to dryness
under reduced pressure. This material was purified by
sublimination (145C, <1 mm), followed by
recrystallization from benzene to give the title
compound (5.4 g, 45~) as pale yellow crystals. mp 149-
150C; lH NMR (CDC13) 7.20 (m, 5H), 4.50 (s, 2H), 3.60
(s, 2H).
2-Am;nomethylphenyl~cetic Ac;~-~Cl
The title compound was prepared by modification of
procedures previously reported in the literature (Naito
et al. (1977) J. Antibiotics, 30: 698). A mixture of 2-
aminomethylphenylacetic acid d-lactam (6.4 g, 44 mmol)
and 21 ml of 6N HCl was heated to reflux for 4 hours.
The reaction mixture was treated with activated carbon
(Norit A), filtered, evaporated to dryness, and the
residual oil triturated with acetone. Filtration
provided the title compound (5.5 g, 62%) as colorless
crystals. mp 168C (dec); 1H NMR (D6-DMSO) 12.65 (br s,
lH), 8.35 (br s, 3H), 7.50 (m, lH), 7.35 (m, 3H), 4.05
(ABq, 2H), 3.80 (s, 2H).
r
2-Aminomethylhenzo;c Ac;~ g-T.~ct~m
--101--
W094/22910 PCT~S94/032~
The title compound was prepared by modification of
procedures previously reported in the literature
(Danishefsky et al. (1975) J. Org. C~em., 40: 796). A
mixture of methyl o-toluate (45 g, 33 mol), N-
bromosuccinimide (57 g, 32 mol), and dibenzoyl peroxide
(0.64 g) in 175 ml of carbon tetrachloride was heated to
reflux for 4 hours. The cooled reaction mixture was
filtered, evaporated to dryness under reduced pressure,
dissolved in 250 ml of methanol, and concentrated
10 ammonium hydroxide (75 ml, 1.11 mol) was added. The
reaction mixture was heated to reflux for 5 hours,
concentrated, filtered, and the solid washed with H2O
followed by ether. This material was purified by
recrystallization from H2O to give the title compound
15 (11.0 g, 26%) as a white solid. mp 150C; lH NMR (CDCl3)
7.90 (d, lH), 7.60 (t, lH), 7.50 (t, 2H), 7.00 (br s,
lH), 4.50 (s, 2H).
2-Aminomethylhenzoic Acid-HCl
The title compound was prepared using the general
procedure described above for 2-aminomethylphenylacetic
acid-HCl. The lactam (3.5 g, 26 mmol) was converted to
the title compound (2.4 g, 50%) as colorless crystals.
mp 233C (dec); lH NMR (D6-DMSO) 13.40 (br s, lH), 8.35
25 (br s, 3H), 8.05 (d, lH), 7.60 (m, 3H), 4.35 (br s, 2H).
~lternAtives to ~mh: Other Cyclic Peptide Intermediates
Alternatives to Mamb useful as carbocylic residues R31
in the cyclic peptides of the invention include aminoalkyl-
naphthoic acid and aminoalkyl-tetrahydronaphthoic acid
residues. Representative aminoalkyl-naphthoic acid and
aminoalkyl-tetrahydronaphthoic acid intermediates useful in
the synthesis of cyclic peptides of the present invention are
-102-
WO94/22910 215 9 ~ 7 ~ PCT~S94/03223
.
described below. The synthesis of these intermediates is
outlined below in Scheme 4a.
Scheme 4a
~CO2H EIOH ~--CO2Et AlClaAc~yla
1 H2SO~ bJ 2 EDC 38%
CH,C~-- H~O~ ~ CH,C~co2H
NaOa, N~OH
~ o ~ co2H Ala3,~a
Ha 95% HOC~ 5
~ . o ~ H2/Pd-C ~
HOC~ 72~ HOC~27% J~?<NJH2
H2So~ 49% ~ Ha
~ 1 .)TMSCN t Znl2 in Ben~ene ,0
"J~ J 2.)POCb/Pyndine; rdl~ O ~
u~ oc ~ oc ~
9 O / 11 CN
1.) H~Pd-C
Dioxane 2.) N~-BOC
~, l.)~C 73% ~ 3.) SaponUy
HOC~J ~ec~ ~J ~CiQ
14 CH~ E1CC 13 CN HOC
12 H CH~M:I tBCC
The title compound was prepared according to a
modification of standard procedures previously reported
in the literature (Earnest, I., Kalvoda, J., Rihs, G.,
and Mutter, M., Tett. Lett., Vol. 31, No. 28, pp 4011-
4014, 1990).
-103-
WO94/22910~ ~ a PCT~S94/03223
8-A~;no-5 6 7.8-tetr~hy~ro-2-n~phthoic Ac;d
Hy~rochlor;~e (8)
As shown below in Scheme 4a, 4-phenylbutyric acid
(1) was converted to the ethyl ester (2) which was
acylated via aluminum chloride and acetylchloride to
give 4-acetylphenylbutyric acid ethyl ester (3). This
ester was subjected to saponification to give 4-
acetylphenylbutyric acid (4). Subsequently, the acetyl
group was oxidized to give 4-carboxyphenylbutyric acid
(5) which was converted to the 1-tetralin-7-carboxylic
acid (6) using aluminum chloride in a Friedel-Crafts
cyclization with resonably high yield. At that point,
the tetralone was split into two portions and some was
converted to the oxime (7) using sodium acetate and
hydroxylamine hydrochloride.The oxime was subjected to
hydrogenolysis to give the racemic mixture of 8-amino-
5,6,7,8-tetrahydro-2-naphthoic acid as the hydrochloride
(8) for use as an intermediate for incorporation into
the cyclic peptide.
P~rt A - A solution of 4-phenylbutyric acid (50.0 g, 0.3
mol) in ethanol (140 mL) with concentrated sulfuric acid
(0.53 mL) was stirred at reflux over 5 hours. The cooled
solution was poured into ice water and extracted with
ethyl acetate. The combined organic layers were
backwashed with brine, dried over anhydrous magnesium
sulfate and evaporated to dryness under reduced pressure
to give 4-phenylbutyric acid ethyl ester (56.07 g, 0.29
mol, 97%) as a yellow liquid. 1H NMR (CDC13) d 7.3-7.1
(m, 5H), 4.1 (q, 2H, J=7.1 Hz), 2.7 (t, 2H, J=7.7 Hz),
2.3 (t, 2H, J=7.5 Hz), 1.95 (quintet, 2H, J=7.5 Hz),
1.25 (t, 3H, J=7.1 Hz).
P~rt R - To a solution of aluminum chloride (153 g, 1.15
mol), and acetyl chloride (38.5 mL, 42.5 g, 0.54 mol) in
--10~--
WO94122910 2 1 5 9 0 7 ~ PCT~S94/032~
.
dichloromethane (1500 mL) was added, dropwise, a
solution of 4-phenylbutyric acid ethyl ester (S0.0 g,
0.26 mol) in dichloromethane (500 mL). All was stirred
at ambient temperature for 15 minutes. The solution was
poured into cold concentrated hydrochloric acid (2000
mL) and then extracted with dichloromethane. The
combined organic layers were backwashed with brine,
dried over anhydrous magnesium sulfate and evaporated to
dryness under reduced pressure to give 4-
acetylphenylbutyric acid ethyl ester (53.23 g, 0.23 mol,88%) as a dark yellow liquid. lH NMR (CDC13) d 7.9 (d,
2H, J=8.1 Hz), 7.25 (d, 2H, J=8.4 Hz), 4.1 (q, 2H, J=7.1
Hz), 2.75 (t, 2H, J=7.6 Hz), 2.6 (s, 3H), 2.35 (t, 2H,
J=7.6 Hz), 2.0 (quintet, 2H, J=7.5 Hz), 1.25 (t, 3H,
J=7.1 Hz).
PArt C -To a solution of 4-acetylphenylbutyric acid
ethyl ester (50.0 g, 0.21 mol) in ethanol (1250 mL) was
added, dropwise, a solution of sodium hydroxide (50.0 g)
20 in water (1250 mL). All was stirred at reflux over 4
hours. The solution was concentrated to half volume and
then acidified to a pH equal to 1.0 using hydrochloric
acid (lN). The resulting precipitate was collected and
washed with water to give 4-acetylphenylbutyric acid
25 (53.76 g, 0.26 mol, 99%) as a white solid. mp = 50-52C;
lH NMR (CDC13) d 7.9 (d, 2H, J=8.1 Hz), 7.25 (d, 2H,
J=9.1 Hz), 2.75 (t, 2H, J-7.7 Hz), 2.6 (s, 3H), 2.4 (t,
2H, J=7.3 Hz), 2.0 (quintet, 2H, J=7.4 Hz).
P~rt D -To a solution of sodium hypochlorite (330 mL,
17.32 g, 0.234 mol) in a solution of sodium hydroxide
(50%, 172 mL), warmed to 55C, was added, portionwise as
a solid, 4-acetylphenylbutyric acid (16.0 g, 0.078 mol)
~ while keeping the temperature between 60-70C. All was
stirred at 55C over 20 hours. The cooled solution was
quenched by the dropwise addition of a solution of
-105-
WO94/22910 PCT~S94/032~
2~,,S9~
sodium bisulfite (25%, 330 mL). The mixture was then
transferred to a beaker and acidified by the careful
addition of concentrated hydrochloric acid. The
resulting solid was collected, washed with water and
dried, then triturated sequentially with chlorobutane
and hexane to give 9-carboxyphenylbutyric acid (15.31 g,
0.074 mol, 95%) as a white solid. mp = 190-195C; lH NMR
(DMSO) d 12.55 (bs, lH), 8.1 (s, lH), 7.85 (d, 2H, J=8.1
Hz), 7.3 (d, 2H, J=8.1 Hz), 2.7 (t, 2H, J=7.5 Hz), 2.2
(t, 2H, J=7.4 Hz), 1.8 (quintet, 2H, J=7.5 Hz).
PArt ~ - A mixture of 4-carboxyphenylbutyric acid (10.40
g, 0.05 mol), aluminum chloride (33.34 g, 0.25 mol) and
sodium chloride (2.90 g, 0.05 mol) was heated with
continual stirring to 190C over 30 minutes. As the
mixture cooled to 60C, cold hydrochloric acid (lN, 250
mL) was carefully added. The mixture was extracted with
dichloromethane. The combined organic layers were
backwashed with dilute hydrochloric acid and water,
dried over anhydrous magnesium sulfate and evaporated to
dryness under reduced pressure. The resulting solid was
triturated with chlorobutane to give l-tetralon-7-
carboxylic acid (9.59 g, 0.05 mol, 100%) as a brown
solid. mp = 210-215C; lH NMR (DMSO) d 8.4 (s, lH), 8.1
(d, 2H, J=8.0 Hz), 7.5 (d, lH, J=7.9 Hz), 3.0 (t, 2H,
J=6.0 Hz), 2.65 (t, 2H, J=6.6 Hz), 2.1 (quintet, 2H,
J=6.3 Hz).
P~rt F - A solution of l-tetralon-7-carboxylic acid (1.0
g, 0.0053 mol) and sodium acetate (1.93 g, 0.024 mol)
and hydroxylamine hydrochloride (1.11 g, 0.016 mol) in a
mixture of methanol and water (1:1, 15 mL) was stirred
at reflux over 4 hours. The mixture was cooled and then
added was more water (50 mL). The solid was collected,
washed with water and dried, then triturated with hexane
to give l-tetralonoxime-7-carboxylic acid (0.78 g,
-106-
WO94/22910 PCT~S94/032~
2I~9~ 7~
0.0038 mol, 72%) as a white solid. mp = 205-215C; 1H
NMR (DMSO) d 11.3 (s, 2H), 8.4 (s, lH), 7.8 (d, lH,
J=7.7 Hz), 7.3 (d, lH, J=7.7 Hz), 2.8 (t, 2H, J=5.9 Hz),
2.7 (d, 2H, J=6.6 Hz), 1.9-1.7 (m, 2H).
,.
P~rt G - A mixture of 1-tetralonoxime-7-carboxylic acid
(0.75 g, 0.0037 mol) in methanol (25 mL) with
concentrated hydrochloric acid (0.54 mL, 0.20 g, 0.0056
mol) and palladium on carbon catalyst (0.10 g, 5% Pd/C)
was shaken for 20 hours at ambient temperature under an
atmosphere of hydrogen (60 psi). The reaction mixture
was filtered over Celite@ and washed with methanol. The
filtrate was evaporated to dryness under reduced
pressure and the residue was purified by flash
chromatography using hexane:ethyl acetate::1:1 to give
the racemic mixture of 8-amino-5,6,7,8-tetrahydro-2-
naphthoic acid hydrochloride (0.225 g, 0.001 mol. 27%)
as a white solid. mp = 289-291C; lH NMR (DMSO) d 8.55
(bs, 3H), 8.2-8.1 (m, lH), 7.85-7.8 (m, lH), 7.35-7.25
(m, lH), 4.5 (m, lH), 2.9-2.8 (m, 2H), 2.1-1.9 (m, 3H),
1.85-1.7 (m, lH).
-107-
WO941Z2910 PCT~S94/032~
.
~s~ Q
N-(BOC)-8-Aminomethyl-5 6 7 8-tetrahydro-2-naphthoic
Acid ~12)
As shown below in Scheme 4a, the remaining
tetralone was then converted to the methyl ester t9).
Using a procedure from Gregory, G.B. and Johnson, A.L,
JOC, 1990, 55, 1479, the tetralone methyl ester (9) was
converted, first, to the cyanohydrin by treatment with
trimethylsilylcyanide and zinc iodide and then, via the
in situ dehydration with phosphorous oxychloride in
pyridine, to the methyl 8-cyano-5,6-dihydro-2-naphthoate
(11). This naphthoate was divided into two portions and
some was subjected to hydrogenolysis, N-BOC-protection
and saponification to give N-(BOC)-8-aminomethyl-
5,6,7,8-tetrahydro-2-naphthoic acid (12) as an
intermediate for incorporation into the cyclic peptide.
P~rt A - A mixture of 1-tetralon-7-carboxylic acid (7.0
g, 0.037 mol) in methanol (13.6 mL, 10.8 g, 0.30 mol)
with a catalytic amount of hydrochloriic acid (0.07 mL,
0.12 g, 0.0012 mol) was stirred at reflux over 5 hours.
The cooled reaction mixture was poured into ice water
and extracted with ethyl acetate. The combined organic
layers were backwashed with water and brine, dried over
anhydrous magnesium sulfate and evaporated to dryness
under reduced pressure. The resulting solid was purified
by flash chromatography using hexane:ethyl
acetate::75:25. The resulting solid was triturated with
hexane to give 1-tetralon-7-carboxylic acid methyl ester
(3.61 g, 0.018 mol, 49%) as a yellow solid. mp = 170-
172C; lH NMR (CDCl3) d 8.7 (s, lH), 8.15 (d, lH, J=8.1
Hz), 7.35 (d, lH, J=8.1 Hz), 3.95 (s, 3H), 3.05 (d, 2H,
J=6.1 Hz), 2.7 (t, 2H, J=6.4 Hz), 2.15 (quintet, 2H,
J=6.2 Hz).
-108-
WO94/22910 PCT~S94/032~
2l~sa70
.
P~rt B - A solution of 1-tetralon-7-carboxylic acid
methyl ester (3.50 g, 0.017 mol), trimethylsilylcyanide
(1.98 g, 0.02 mol) and zinc iodide (0.10 g) in benzene
(20 mL) was stirred at ambient temperature over 15
hours. Then added, sequentially and dropwise, was
pyridine (20 mL) and phosphorous oxychloride (4.0 mL,
6.55 g, 0.0425 mol). The reaction mixture was stirred at
reflux over 1 hour then evaporated to dryness under
10 reduced pressure. The residue was taken up in
chloroform, backwashed with water, dried over anhydrous
magnesium sulfate and evaporated to dryness under
reduced pressure to give methyl 8-cyano-5,6-dihydro-2-
naphthoate (1.70 g, 0.008 mol, 47%) as a yellow solid.
mp = 73-75C; lH NMR (CDC13) d 8.0-7.9 (m, lH), 7.3-7.2
(m, lH), 6.95 (t, lH, J-4.8 Hz), 3.95 (s, 3H), 2.9 (t,
2H, J=8.3 Hz), 2.6-2.4 (m, 3H)
P~rt C - A mixture of methyl 8-cyano-5,6-dihydro-2-
naphthoate (0.80 g, 0.0038 mol) in methanol (25 mL) with
concentrated hydrochloric acid (0.56 mL) and palladium
on carbon catalyst (0.40 g, 5% Pd/C) was shaken for 20
hours at ambient temperature under an atmosphere of
hydrogen (50 psi). The reaction mixture was filtered
over Celite and washed with methanol. The filtrate was
evaporated to dryness under reduced pressure and the
residue was triturated with hexane to give the racemic
mixture of methyl 8-aminomethyl-5,6,7,8-tetrahydro-2-
naphthoate (0.80 g, 0.0037 mol, 97%) as a white solid.
mp = 172-179C; lH NMR (DMSO) d 8.2-8.0 (m, 4H), 7.9-7.7
(m, 6H), 7.5-7.2 (m, 4H), 3.9-3.8 (m, 7H), 3.3-2.7(m,
lOH), 2.0-1.6 (m, 8H).
p~rt D - A solution of methyl 8-aminomethyl-5,6,7,8-
tetrahydro-2-naphthoate (0.78 g, 0.0036 mol) and
triethylamine (0.55 mL, 0.40 g, 0.004 mol) in aqueous
--1 0 9--
W094/22910 PCT~S94/032~
tetrahydrofurrn (50~, 75 mL) was added, portionwise as a
solid, 2-(tert-butoxycarbonyloxyimino)-2-
phenylacetonitrile (0.99 g, 0.004 mol). All was stirred
at ambient temperature over 3 hours. The solution was
concentrated to half volume and extracted with
diethylether. The aqueous layer was then acidified to a
pH of 1.0 using hydrochloric acid (lN) and then extraced
with ethyl acetate. The combined organic layers were
dried over anhydrous magnesium sulfate and evaporated to
dryness under reduced pressure. The residue was purified
by flash chromatography using hexane:ethyl acetate::8:2
to give methyl N-(BOC)-8-aminomethyl-5,6,7,8-tetrahydro-
2-naphthoate (0.54 g, 0.0017 mol, 47%) as a white solid.
mp = 72-80C; 1H NMR (DMSO) d 13.8 (s, lH), 7.8-7.65 (m,
3H), 7.6-7.5 (m, 3H), 7.25-7.20 (m, lH), 7.15-7.05 (m,
lH), 3.9-3.8 (m, lH), 3.2-2.8 (m, 4H), 1.8-1.6 (m, 3H),
1.4 (s, 6H).
P~rt F. - To a solution of methyl N-(BOC)-8-aminomethyl-
5,6,7,8-tetrahydro-2-naphthoate (0.50 g, 0.0016 mol) in
ethanol (12.5 mL) was added, dropwise, a solution of
sodium hydroxide (0.50 g) in water (12.5 mL). All was
stirred a reflux over 4 hours. The reaction mixture was
concentrated to half volume and then acidified to a pH
equal to 1.0 using hydrochloric acid (lN). The residue
was puified by flash chromatography using a gradient of
hexane:ethyl acetate::1:1 to ethyl acetate to ethyl
acetate: methanol::9:1 to give the racemic mixture of
the title compound, N-(BOC)-2-aminomethyl-5,6,7,8-
tetrahydro-2-naphthoic acid (0.19 g, 0.00062 mol, 39%)
as a white solid. mp = 172-176C; 1H NMR (DMSO) d 7.8
(s, lH), 7.65 (d, lH, J=8.1 Hz), 7.15 (d, lH, J=8.1 Hz),
7.1-7.0 (m, lH), 3.2-3.1 (m, 2H), 3.0-2.7 (m, 4H), 1.8-
1.6 (m, 4H), 1.4 (s, 9H).
--1 1 0--
W094l22910 PCT~S94/032~
~ 21~9~7~
N- (ROC) -8-~m; nomethyl-2-nAphthol c ~ci ~ ( 14 )
The remaining naphthoate (11) was treated with 2,3-
dichloro-5,6-dicyano-1,4-benzoquinone (DDQ) in dioxane
to aromatize the adjacent ring to give the methyl 8-
cyano-2-naphthoate (13). Then, the nitrile was reduced
via hydrogentation and the methyl ester saponified to
the carboxylic acid. This acid was then N-BOC-protected
to give N-(BOC)-8-aminomethyl-2-naphthoic acid (14) as
an intermediate for incorporation into the cyclic
peptide.
P~rt A - A solution of methyl 8-cyano-5,6-dihydro-2-
naphthoate (1.0 g, 0.0047 mol) and 2,3-dichloro-5,6-
dicyano-1,4-benzoquinone (1.07 g, 0.0047 mol) in dioxane
(50 mL) was stirred at 120C over 16 hours. The reaction
mixture was poured into ice water and extracted with
ethyl acetate. The combined organic layers were dried
over anhydrous magnesium sulfate and evaporated to
dryness under reduced pressure. The residue was purified
by flash chromatography using ethyl acetate to give
methyl 8-cyano-2-naphthoate (0.72 g, 0.0034 mol, 73%) as
a tan solid. mp = 178-182C; lH NMR (CDC13) d 8.95 (s,
lH), 8.3-8.2 (m, lH), 8.15-8.10 (m, lH), 8.0-7.95 (m,
2H), 7.7-7.6 (m, lH), 4.05 (s, lH).
P~rt R - A mixture of methyl 8-cyano-2-naphthoate (1.0
g, 0.0047 mol) in methanol (35 mL) with concentrated
hydrochloric acid (0.69 mL) andpalladium on carbon
catalyst (0.20 g, 5% Pd/C) was shaken for 6 hours at
ambient temperature under anatmosphere of hydrogen (50
psi). The reaction mixture was filtered over celiteQ and
washed with methanol. The filtrate was evaporated to
dryness under reduced pressure and the residue was
triturated with hexane to give methyl 8-aminomethyl-2-
--111--
-
W094l22910 PCT~S94/032~
naphthoate (0.76 g, 0.0035 mol, 75%) as an oil. lH NMR
(DMSO) d 8.75 (s, lH), 8.5 (bs, 2H), 8.2-8.05 (m, 3H),
7.75-7.70 (m, 2H), 4.6 (s, 2H), 3.95 (m, 3H).
P~rt C - To a solution of methyl 8-aminomethyl-2-
naphthoate (0.75 g, 0.0035 mol) in dry tetrahydrofuran
(50 mL), cooled to 0C, was added a solution of lithium
hydroxide (0.5 M, 5.83 mL). All was stirred at ambient
temperature over 20 hours. Another aliquot of lithium
hydroxide was added and all was stirred for an
additional 20 hours. The solid was collected and the
filtrate was evaporated to dryness under reduced
pressure. The solids were triturated with diethyl ether
to give 8-aminomethyl-2-naphthoic acid (0.67 g, 0.0033
15 mol, 95%) as a white solid. mp = 223-225C; lH NMR
(DMSO) d 8.6 (s, lH), 8.1-7.9 (m, lH), 7.8-7.7 (m, 4H),
7.55-7.5 (m, lH), 7,45-7.35 (m, 2H), 4.2 (s, 2H).
P~rt D - A solution of 8-aminomethyl-2-naphthoic acid
20 (0.50 g, 0.00025 mol) and triethylamine (0.038 mL, 0.028
g, 0.000275 mol) in aqueous tetrahydrofuran (50%, 5 mL)
was added, portionwise as a solid, 2-(tert-
butoxycarbonyloxyimino)-2-phenylacetonitrile (0.068 g,
0.000275 mol). All was stirred at ambient temperature
over 5 hours. The solution was concentrated to half
volume and extracted with diethylether. The aqueous
layer was then acidified to a pH of 1.0 using
hydrochloric acid (lN) and then extraced with ethyl
acetate. The combined organic layers were dried over
anhydrous magnesium sulfate and evaporated to dryness
under reduced pressure to give the title compound, N-
(BOC)-8-aminomethyl-2-naphthoic acid (0.050 g, 0.00017
mol) as a white solid. mp = 190-191C; lH NMR (DMSO) d
13.1 (bs, lH), 8.8 (s, lH), 8.0 (q, 2H, J=7.9 Hz), 7.9
35 (d, lH, J=8.1 Hz), 7.6 (t, lH, J=7.5 Hz), 7.65-7.55 (m,
2H), 4.6 (d, 2H, J=5.5 Hz), 1.4 (s, 9H).
-112-
WO94122910 PCT~S94/032~
21S907~
Synthesis of Cyclic Pe~t;~es
t-Butyloxycarbonyl-3-aminomethylbenzoic acid (Boc-
Mamb) is coupled to oxime resin by a modification of the
~ method described by DeGrado and Kaiser (1980) J. Org.
Chem. 45, 1295 using 1 equivalent of the 3-
aminomethylbenzoic acid (with respect to the
substitution level of the resin), 1 equivalent of HBTU,
and 3 equivalent of NMM. Alternatively, Boc-Mamb (l
equivalent) may be coupled to the oxime resin using 1
equivalent each of DCC and DMAP in methylene chloride.
Coupling times range from 15 to 96 hours. The
substitution level is then determined using either the
picric acid test (Sarin, Kent, Tam, and Merrifield,
(1981) Anal. Biochem. 117, 145-157) or the quantitative
ninhydrin assay (Gisin (1972) Anal. Chim. Acta 58, 248-
249). Unreacted oxime groups are blocked using 0.5 M
trimethylacetylchloride / 0.5 M diisopropylethylamine in
DMF for 2 hours. Deprotection of the Boc protecting
group is accomplished using 25% TFA in DCM for 30
minutes. The remaining amino acids or amino acid
derivatives are coupled using between a two and ten fold
excess (based on the loading of the first amino acid or
amino acid derivative) of the appropriate amino acid or
amino acid derivatives and HBTU in approximately 8 ml of
DMF. The resin is then neutralized in situ using 3 eq.
of NMM (based on the amount of amino acid used) and the
coupling times range from 1 hour to several days. The
completeness of coupling is monitored by qualitative
ninhydrin assay, or picric acid assay in cases where the
amino acid was coupled to a secondary amine. Amino
acids are recoupled if necessary based on these results.
After the linear peptide had been assembled, the N-
terminal Boc group is removed by treatment with 25% TFA
-113-
W094l22910 PCT~S94/032~
in DCM for 30 minutes. The resin is then neutralized by
treatment with 10% DIEA in DCM. Cyclization with
concomitant cleavage of the peptide is accomplished
using the method of Osapay and Taylor ((1990) J. Am.
5 Chem. Soc., 112, 6046) by suspending the resin in
approximately 10 ml/g of DMF, adding one equivalent of
HOAc (based on the loading of the first amino acid), and
stirring at 50-60 C for 60 to 72 hours. Following
filtration through a scintered glass funnel, the DMF
filtrate is evaporated, redissolved in HOAc or 1:1
acetonitrile: H2O, and lyophilized to obtain protected,
cyclized material. Alternatively, the material may be
dissolved in methanol and precipitated with ether to
obtain the protected, cyclized material. This is then
15 treated using standard procedures with anhydrous
hydrogen fluoride (Stewart and Young (1984) "Solid Phase
Peptide Synthesis", 2nd. edition, Pierce Chemical Co.,
85) containing 1 ml/g m-cresol or anisole as scavenger
at 0 C for 20 to 60 minutes to remove side chain
protecting groups. The crude product may be purified by
reversed-phase HPLC using a 2.5 cm preparative Vydac C18
column with a linear acetonitrile gradient containing
0.1% TFA to produce pure cyclized material. The
following N-a-Boc-protected amino acids may be used for
the syntheses: Boc-Arg(Tos), Boc-N-a-MeArg(Tos), Boc-
Gly, Boc-Asp(OcHex), Boc-3-aminomethyl-4-iodo-benzoic
acid, Boc-D-Ile, Boc-NMeAsp(OcHex), Boc-NMe-Mamb, Boc-D-
Phg, Boc-D-Asp(OBzl), Boc-L-Asp(OcHex), Boc-aMe-
Asp(OcHex), Boc-bMe-Asp(OcHex), Boc-L-Ala, Boc-L-Pro,
Boc-D-Nle, Boc-D-Leu, Boc-D-Val, Boc-D-2-aminobutyric
acid (Boc-D-Abu), Boc-Phe, Boc-D-Ser(Bzl), Boc-D-Ala,
Boc-3-aminomethylbenzoic acid (Boc-Mamb), Boc-D-Lys(2-
ClZ), Boc-~-Ala, Boc-D-Pro, Boc-D-Phe, Boc-D-
Tyr(Cl2Bzl), Boc-NMe-Amf(CBZ), Boc-aminotetralin-
carboxylic acid, Boc-aminomethylnaphthoic acid, Boc-4-
aminomethylbenzoic acid, or Boc-NMeGly.
-114-
WO94122910 21 5 ~ ~ ~ o PCT~S94/032~
The synthesis of the compounds of the invention is
further exemplified in PCT Patent Application
International Publication Number WO93/07170 (Publication
Date April 15, 1993) and as set forth below. The Tables
below set forth representative compounds of the present
invention.
F,~Am~1 10 cyclo-(Abu-NMeArg-Gly-Asp-Mamb); the compound of formula
(II) wherein J = Abu, K = NMeArg,
L = Gly, M = Asp, Rl = H, R2 = H
The title compound was prepared using the general
procedure described for cyclo-(D-Val-NMeArg-Gly-Asp-
Mamb) (Example 4). The DCC/DMAP method was used for
attachment of Boc-Mamb to the oxime resin. TBTU was
used as the coupling reagent. The peptide was prepared
on a 0.596 mmol scale to give the protected cyclic
peptide (182 mg,38.4%). The peptide (176 mg) and 0.176
mL of anisole were treated with anhydrous hydrogen
fluoride at 0 C for 20 minutes. The crude material was
precipitated with ether, redissolved in aqueous
acetonitrile, and lyophilized to generate the title
compound (116 mg; 90.4%; calculated as the fluoride
salt). Purification was accomplished by reversed-phase
HPLC on a preparative Vydac C18 column (2.5 cm) using a
0.45%/ min. gradient of 9 to 27~ acetonitrile containing
0.1% TFA and then lyophilized to give the TFA salt of
the title compound as a fluffy white solid (1.92%
recovery, overall yield 0.574%); FAB-MS: [M+H] = 561.39.
Fx~le 4
Cryst~ z~tion of the Com~ound of ~xA~le 4 ~nd the
Prep~rAtion of S~lt Form~ of the Compound of ~x~m~le 4
-115-
W094/22910 PCT~S94/03223
.
~,~9
It has been discovered that the compounds of the
present invention may be isolated by crystallization of
the compound from organic and aqueous solvents.
The zwitterion of Example 4 was converted to the
mesyl (methanesulfonate) salt of Example 4 (Example 4
(methane-sulfonate)) by refluxing the zwitterion with
stirring in isopropanol at 25 mg/ml and slowly adding a
solution of l.0 molar equivalent methanesulfonic acid
(correcting for the water content of the zwitterion)
dissolved in isopropanol. The heat was turned off and
the solution cooled to 5C in an ice bath. After
stirring 1 hour, the solution was filtered and the solid
rinsed three times with cold isopropanol and dried under
vacuum to constant weight.
The following salts of the compound of Example 4
were prepared using the same procedure, by adding 1.0
equivalent of the appropriate acid:
Example 4 (biphenylsulfonate):
zwitterion + 1.0 equivalent biphenylsulfonic acid.
Example 4 (a-naphthalenesulfonate):
zwitterion + 1.0 equiv. ~-naphthalenesulfonic acid.
Example 4 (~-naphthalenesulfonate):
zwitterion + 1.0 equiv. ~-naphthalenesulfonic acid.
Example 4 (benzenesulfonate):
zwitterion + 1.0 equiv. benezene-sulfonic acid.
Example 4 (p-toluenesulfonate):
zwitterion + 1.0 equiv. p-toluene-sulfonic acid.
-116-
WO94/22910 PCT~S94/03223
21~7~
.
The following salts of the compound of Example 4
were prepared by crystallization of the compound from
aqueous systems.
Example 4 (sulfate):
10 mg amorphous Example 4 (made by lyophilizing the
zwitterion from a solution of 2 molar equivalents of
acetic acid in water) dissolved per ml 1 N H2SO4, pH
adjusted to 2.5. On standing at room temperature, a
precipitate formed. This was filtered through a
sintered glass funnel and dried under vacuum to constant
weight.
Example 4 (methanesulfonate (mesyl)):
100 mg amorphous DMP728 dissolved per ml water + 1.2
molar equiv. methanesulfonic acid (this was obtained as
a 4M aqueous solution). On standing at room
temperature, a large flat crystal was formed.
Example 4 (benzenesulfonate):
100 mg zwitterion dissolved per ml water + 1.2 equiv.
benzenesulfonic acid added. On standing at room
temeprature, a precipitate formed. This was filtered
through a sintered glass funnel, rinsed with a small
volume of isopropanol, and dried under vacuum to
constant weight.
Example 4 (p-toluenesulfonate):
100 mg zwitterion dissolved per ml water + 1.2 molar
equiv. toluenesulfonic acid added. On standing at room
temperature, a precipitate formed. This was filtered
through a sintered glass funnel and dried under vacuum
to constant weight.
F.xample 17
-117-
WO94/22910 PCT~S94/032~
~'9~ --
cyclo-(D-Met-NMeArg-Gly-Asp-Mamb); the compound of
formula (II) wherein J = D-Met, K = NMeArg, L = Gly, M =
Asp, Rl = H, R2 = H
The title compound was prepared using the general
procedure described for cyclo-(D-Val-NMeArg-Gly-Asp-
Mamb) (Example 4). The DCC/DMAP method was used for the
attachment of Boc-Mamb to the resin. The peptide was
prepared on a 0.179 mmol scale to give the protected
cyclic peptide (105 mg, 69.7%). The peptide (105 mg)
and 0.105 mL of anisole were treated with anhydrous
hydrogen fluoride at 0 C for 20 minutes. The crude
material was precipitated with ether, redissolved in
aqueous acetonitrile, and lyophilized to generate the
title compound (72 mg; 92.3% yieldi calculated as the
fluoride salt). Purification was accomplished by
reversed-phase HPLC on a preparative Vydac C18 column
(2.5 cm) using a 0.23%/ min. gradient of 14.4 to 23.4%
acetonitrile containing 0.1% TFA and then lyophilized to
give the TFA salt of the title compound as a fluffy
white solid (13.2% recovery, overall yield 7.4%);
FAB-MS: [M+H~ = 607.3.
Fxam~le 401
cyclo-(D-Abu-NMeArg-Gly-D-Asp-Mamb); compound of
formula tII) wherein J = D-Abu, K = NMeArg, L = Gly, M =
D-Asp, R1 = H, R2 = H
The title compound was prepared using the general
procedure described for cyclo-(D-Val-NMeArg-Gly-Asp-
Mamb) (example 4). The DCC/DMAP method was used for
attachment of Boc-Mamb to the oxime resin. TBTU was
used as the coupling reagent. The peptide was prepared
on a 0.596 mmol scale to give the protected cyclic
peptide (273 mg, 57.6%). The peptide ~263 mg) and 0.263
-118-
W094/22910 ~ 0 7~ PCT~S94/032~
.
mL of anisole were treated with anhydrous hydrogen
fluoride at 0 C for 20 minutes. The crude material was
precipitated with ether, redissolved in aqueous
acetonitrile, and lyophilized to generate the title
5 compound (218 mg; greater than quantitative yield;
calculated as the fluoride salt). Purification was
accomplished by reversed-phase HPLC on a preparative
Vydac C18 column (2.5 cm) using a 0.23%/ min. gradient
of 10.8 to 19.8% acetonitrile containing 0.1% TFA and
10 then lyophilized to give the TFA salt of the title
compound as a fluffy white solid (40.4% recovery,
overall yield 21.9%); FAB-MS: [M+H] = 561.37.
F.x~mnle 402
cyclo-(D-Abu-D-NMeArg-Gly-Asp-Mamb); the compound of
formula (II) J = D-Abu, K = D-NMeArg, L = Gly, M = Asp,
R1 - H, R2 = H
The title compound was prepared using the general
procedure described for cyclo-(D-Val-NMeArg-Gly-Asp-
Mamb) (example 4). The DCC/DMAP method was used for
attachment of Boc-Mamb to the oxime resin. TBTU was
used as the coupling reagent. The peptide was prepared
on a 0.596 mmol scale to give the protected cyclic
2S peptide (241 mg, 50.8%). The peptide (235 mg) and 0.235
mL of anisole were treated with anhydrous hydrogen
fluoride at 0 C for 20 minutes. The crude material was
precipitated with ether, redissolved in aqueous
acetonitrile, and lyophilized to generate the title
compound (168 mg; 98.3%; calculated as the fluoride
salt). Purification was accomplished by reversed-phase
HPLC on a preparative Vydac C18 column (2.5 cm) using a
0.23%/ min. gradient of 12.6 to 21.6% acetonitrile
containing 0.1% TFA and then lyophilized to give the TFA
salt of the title compound as a fluffy white solid (2.3%
recovery, overall yield 0.99%); FAB-MS: [M+H] = 561.36.
--119--
WO94/22910 PCTNS94/032
~.xam~le 403
Cyclo-(D-Ala-p-guanidinyl-Phe-Gly-Asp-Mamb);
the compound of formula (II) wherein J = D-Ala, K = p-
5guanidinyl-Phe, L = Gly, M = Asp R1 = H, R2 = H
NH,
NH,~NH ~F NH ~NH ~F
N~C(rNH)SO~H N'l NH
~\ DMAP~ EtOH
Dissolved 25 mg (38.3 ~moles) of cyclo-(D-Ala-p-
amino-Phe-Gly-Asp-Mamb) (TFA salt), 14.3 mg (114.9
umoles) formamidine sulfonic acid, and 18.7 mg (153.2
umoles) of 4-dimethyl-aminopyridine in 5 ml of ethanol
in a 10 ml round bottom flask. Refluxed the mixture for
3 hours, then added an additional 14.3 mg of formamidine
sulfonic acid and 18.7 mg of 4-dimethyl-aminopyridine.
After refluxing for an additional 3 hours, the reaction
was found to be ~75% complete by reversed-phase HPLC.
The ethanol was evaporated under reduced pressure, and
the residue was purified on a preparative Vydac C18
column (2.5 cm) using a 0.45%/min. gradient of 0 to 18%
acetonitrile containing 0.1% TFA.
Lyophilization afforded the TFA salt of the title
compound as a white solid (28% recovery), overall yield
26.4%); FAB-MS: [M+H] = 581.30.
Fx~le 404
cyclo-(D-Abu-(DiNMe,guanidinyl-Orn)-Gly-Asp-Mamb); the
compound of formula (II) wherein J = D-Abu, K =
diNMe,guanidinyl-Orn , L = Gly, D = Asp, Rl = H, R2 = H
-120-
W094/22910 PCT~S94tO32~
2159~
o ~
~ \F NH2-C(-NH)503H M~ --\F
~ \ ~ Lo
~ \DIAAP, EtOH \ <
Dissolved 10.53 mg (16.3 ~moles) of cyclo-(D-Abu-
diNMeOrn-Gly-Asp-Mamb) (TFA salt), 6.08 mg (48.99
5 umoles) formamidine sulfonic acid, and 8.00 mg (65.57
umoles) of 4-dimethyl-aminopyridine in 2.5 ml of ethanol
in a 10 ml round bottom flask. Refluxed the mixture for
2 hours and then stirred at room temperature overnight.
Refluxed for one hour, added an additional 6.08 mg of
formamidine sulfonic acid and 8.00 mg of 4-
dimethylaminopyridine and then refluxed for an
additional 2 hours. Evaporated the ethanol under
reduced pressure and purified the residue on a
preparative Vydac C18 column (2.5 cm) using a 0.45%/min.
gradient of 3.6 to 18% acetonitrile containing 0.1% TFA.
Lyophilization afforded the TFA salt of the title
compound as a white solid (57.2% recovery), overall
yield 53.5%); FAB-MS: [M+H] = 575.34.
F.~m~l es 405
cyclo-(D-Abu-Di-NMeLys-Gly-Asp-Mamb); the compound of
formula (II) wherein J = D-Abu, K = Di-NMeLys, L = Gly,
M = Asp, Rl = H, R2 = H
cyclo-(D-Abu-NMeLys-Gly-Asp-Mamb); the compound of
formula (II) wherein J = D-Abu, K = NMeLys, L = Gly, M =
Asp, Rl = H, R2 = H
-121-
WO94/22910 ~CT~S94/032~
~g~ --
Di-N-methyl amino acid derivatives may be prepared
using methods which have been described previously
(Olsen, ~. Org. Chem. (1970) 35: 1912) or,
aLternatively, through the use of NaH/CH3I. The mono-
NMe-Lysine amino acid was obtained as a side product
during the synthesis of the corresponding di-NMe-lysine
derivative. The title compounds were prepared using
conventional solution phase peptide chemistry techniques
described previously. Cyclo-(D-Abu-diNMeLys-Gly-Asp-
Mamb) was obtained in 0.31% overall yield, FAB-MS: [M+H]
= 547.3. Cyclo-(D-Abu-NMeLys-Gly-Asp-Mamb) was obtained
in 0.25% overall yield, FAB-MS: [M+H] = 533.3.
~x~le 100a
cyclo-(D-Ahu-NMe~rg-Gly-Asp-3-~m;nomethyl-6-
chlorohenzoic ac;d)
The title compound was prepare by the general
solution-phase procedure described above for cyclo-(D-
Val-NMeArg-Gly-Asp-Mamb), except that 4,4'-
dinitrobenzophenone oxime was employed. The cyclic
peptide (330 mg, 0.40 mmol) was deprotected with excess
HF in the presence of anisole as scavenger. Purification
was accomplished by reversed-phase HPLC on a preparative
Vydac C18 column (2.5 cm) using a 1.0% / minute gradient
of 10 to 38% acetonitrile containing 0.1%
trifluoroacetic acid to give the TFA salt of the title
compound (114 mg, 41%) as a fluffy white solid; lH NMR
(D6-DMSO) 9.00 (d, lH), 8.40 (m, 2H), 7.50 (m, lH), 7.40
(d, lH), 7.30 (m, 2H), 7.15 (s, lH), 7.00 (br s, 4H),
5.15 (dd, lH), 4.65 (q, lH), 4.50 (dd, lH), 4.40 (q,
lH), 4.05 (dd, lH), 3.95 (dd, lH), 3.65 (dd, lH), 3.10
(q, 2H), 3.05 (s, 3H), 2.75 (dd, lH), 2.50 (m, lH), 1.95
(m, lH), 1.75 (m, 2H), 1.60 (m, lH), 1.35 (m, 2H), 0.95
(t, 3H); FAB-MS: [M+H] = 595.4.
-122-
WO941~910 PCT~S94/032~
21~907~
Fx~m~le lOOb
cyclo-(D-~hu-NMe~rg-Gly-A~p-3-~minomethyl-6-;o~ohenzoic
~c; ~)
-
The title compound was prepare by the general
solution-phase procedure described above for cyclo-(D-
Val-NMeArg-Gly-Asp-Mamb), except that 4,4'-
dinitrobenzophenone oxime was employed. The cyclic
peptide (350 mg, 0.38 mmol) was deprotected with excess
HF in the presence of anisole as scavenger. Purification
was accomplished by reversed-phase HPLC on a preparative
Vydac C18 column (2.5 cm) using a 1.0% / minute gradient
of 10 to 38% acetonitrile containing 0.1%
trifluoroacetic acid to give the TFA salt of the title
15 compound (150 mg, 49%) as a fluffy white solid; lH NMR
(D6-DMSO) 8.90 (d, lH), 8.40 (m, 2H), 7.70 (d, lH), 7.50
(m, lH), 7.30 (m, lH), 7.05 (s, lH), 7.00 (d, lH), 7.00
(br s, 4H), 5.15 (dd, lH), 4.65 (q, lH), 4.45 (dd, lH),
4.40 (q, lH), 4.00 (q, lH), 3.90 (q, lH), 3.65 (dd, lH),
20 3.10 (q, 2H), 3.05 (s, 3H), 2.70 (dd, lH), 2.50 (m, lH),
1.95 (m, lH), 1.75 (m, 2H), 1.60 (m, lH), 1.40 (m, 2H),
0.95 (t, 3H); FAB-MS: [M+H] = ~87.3.
F.x~m~le lOOc
cyclo-(D-~hu-NMeAr~-Gly-A~D-3-~m;nomethyl-6-
methylhenzo;c ~c;~)
(the compound of formula (VII) wherein J = D-Abu, K =
NMeArg, L = Gly, M = Asp, R10 = Me)
The title compound was prepare by the general
solution-phase procedure described above for cyclo-(D-
- Val-NMeArg-Gly-Asp-Mamb), except that 4,4'-
dinitrobenzophenone oxime was employed. The cyclic
- peptide (130 mg, 0.16 mmol) was deprotected with excess
HF in the presence of anisole as scavenger. Purification
was accomplished by reversed-phase HPLC on a preparative
-123-
W094/22910 PCT~S94/032~
Vydac C18 column (2.5 cm) using a 1.0% / minute gradient
of 10 to 38~ acetonitrile containing 0.1%
trifluoroacetic acid to give the TFA salt of the title
compound (31 mg, 28%) as a fluffy white solid; lH NMR
(D6-DMSO) 8.70 (d, lH), 8.40 (d, lH), 8.30 (t, lH), 7.50
(m, lH), 7.45 (m, lH), 7.15 (q, 2H), 7.05 (s, lH), 7.00
(br s, 4H), 5.15 (dd, lH), 4.65 (q, lH), 4.45 (m, 2H),
4.00 (m, 2H), 3.65 (dd, lH), 3.10 (q, 2H), 3.05 (s, 3H),
2.75 (dd, lH), 2.50 (m, lH), 2.30 (s, 3H), 2.00 (m, lH),
1.75 (m, 2H), 1.60 (m, lH), 1.35 (m, 2H), 0.95 (t, 3H);
FAB-MS: [M+H] = 575.4.
ReDresentative Prodrugs
Step 1: ~-henzyloxyc~rhonyl-N~-methyl-4-cy~no-T~-2
~m; nohutyr;c ~cid
Z-Gln (28.03 g, 100 mmol) was dissolved in 300 mL
THF in a flask bottle protectected from moisture and to
it was added 100 mL 1.93 M phosgene in toluene (193
mmol). The solution was stirred at room temperature for
2 h and concentrated at 30 C to 200 mL. Water (200 mL)
was added slowly with stirring. After stirring at room
temperature for 2 h, the organic phase was seperated,
and the water phase was extracted with ethyl acetate
twice. The combined organic solution was washed with
brine four times, dried (MgSO4), and concentrated. The
oily product was dried over KOH overnight.
The dried oily product was taken up in 300 mL dry
THF and 49.8 mL (800 mmol) methyl iodide in a flask
bottle protected from moisture and the solution was
cooled in an ice bath. To it was slowly added 10 g
sodium hydride (250 mmol, 60% dispersion in oil). The
mixture was stirred in the ice bath for 1 h and then at
room temperature for 22 h. Ethyl acetate (50 mL) was
added, and after stirring for 10 min, 100 mL water was
added slowly. The solution was acidified with a few
-124-
WO 94/22910 PCT/US94/03223
215907~
.
drops of 4 N HCl to pH8-9 and then concentrated at 30 C
to remove the organic solvents. Water (100 mL) was added
followed by 10 mL 0.1 N sodium thiosulfate, and the
solution was extracted with ether twice. The water layer
5 was cooled in an ice bath and to it was slowly added 4 N
HCl to pH 3 with stirring. The product, which
crystallized during the acidification, was filtered,
washed with water several times, and dried. Yield 26.0 g
(94%). mp 81-83 C. lH-NMR (CDC13): S=2.15 (m, lH); 2.38
(m, lH); 2.42 (m, 2H); 2.96 & 2.98 (2 s, cis ~ trans N-
CH3); 4.62 (m, lH); 4.90 (b, lH); 5.19 (s, 2H); 7.35 (m,
5H).
Step 2: ~-met~yl-4-cy~no-T-2-~m;nohutyric ~c;~l-N-
15 c~rhoxy~nhy~r;~e
To a solution of example 1 (11.05 g, 40 mmol) in 50
mL dry THF cooled in an ice bath was added phosphorus
pentachloride (15 g, 72 mmol) and the mixture was
stirred for 2 h and concentrated to dryness. The residue
20 was triturated with petroleum ether to give a solid
which was filtered, washed with petroleum ether and
dissolved in dry acetonitrile. Insoluble material was
filtered off and the solution was concentrated. The
solid was washed with cold ether and dried. Yield 5.86 g
25 (8796). mp 90-92 C. lH-NMR (CDCl3): ~=2.18 (m, lH); 2.39
(m, lH); 2.60 (m, 2H); 3.02 (s, 3H); 4.28 (m, lH) .
Step 3: N-Roc-D-2-~m;nohutyryl-N9~-methyl-4-cy~rlo-T.-2-
~m;nohlltyryl--s~lyc;ne t-hutyl ester
To a solution of glycine t-butyl ester
hydrochloride (3.68 g, 22 mmol) in 40 mL chloroform and
- 4.84 mL N-methylmorpholine cooled to -40 C was added a
solution of example 2 (3.36 g, 20 mmol) in 20 mL dry
acetonitrile, the solution was stirred at -20 C for 1
35 h, and the solvent was reduced to about 10 mL.
--125--
W094l~910 PCT~S941032
To a solutioA rf N-Boc-D-2-aminobutyric acid
dicyclohexylamine salt (8.08 g, 21 mmol) in 30 mL
chloroform cooled to -10 C was added diphenylphosphinic
chloride (3.91 mL, 20.5 mmol) and the mixture was
stirred at -5 to -10 C for 1 h. To it was added the
above prepared solution (10 mL) followed by 2.42 mL N-
methylmorpholine. The mixture was stirred at 0 to -5 C
for 24 h, and then concentrated. Ethyl acetate was added
and insoluble material was filtered off. The filtrate
was washed with NaHCO3 four times and with brine three
times, dried over MgSO4, and concentrated to a small
amount at which time the product crystallized. Petroleum
ether was added, and after cooling, the solid was
filtered, washed with petroleum ether, and dried. Yield
15 6.2 g (70%). mp 90-92 C. FAB-MS (MH+): Calculated
441.3; Found 441.3.
Step 4: N-Roc-D-2-aminohutyryl-N~-methyl-N~ N~ -
(h;shenzyloxyc~rbonyl)-T-~rginyl-glycine t-hutyl ester
The compound of Step 3 (4.63 g, 10.5 mmol) was
dissolved in 70 mL methanol in a Parr bottle and to it
was added a cold solution of 1.2 mL concentrated
hydrochloric acid (38%) in 10 mL methanol followed by
200 mg platinum(IV) oxide. The mixture was hydrogenated
at 55 psi for 1 h, the catalyst was filtered off, and
2.09 mL (15 mmol) triethylamine was added. The solvent
was removed under reduced pressure and the residue was
taken up in 20 mL THF. To it was added N, N'-
bisbenzyloxycarbonyl-S-methylisothiourea (3.58 g, 10
mmol) followed by 2.09 mL (15 mmol) triethylamine. The
mixture was stirred overnight during which time the
bottle was evacuated several times to remove the
byproduct methanethiol. Ethyl acetate was added, and the
solution was washed with 1% citric acid, brine, 5
NaHCO3 and brine, dried (MgSO4), and concentrated.
Crystallization from ethyl ether-petroleum ether gave
-126-
W094/~910 PCT~S94/032~
21~9~7~
,.
7.2 g (95%) product. FAB-MS (MH+): Calculated 755.4;
Found 755.4.
Step 5: D-2-~m; nohutyryl-N~-methyl-N~J N~ _
(hishenzyloxyc~rhonyl)-T-~r~inyl-glyc;ne TFA s~lt
A solution of the compound of Step 4 (9 g, 11.9
mmol) in 90 mL 50% TFA in methylene chloride was stirred
at room temperature for 2 h and the solution was
concentrated at 30 C. Cold ether was added, and after
standing, the solid was filtered, washed with ether, and
dried. Yield 8.4 g (99%). FAB-MS (MH+): Calculated
599.3; Found 599.3.
Step 6: 3- (~m; nomethyl)henzo;c ~c;~ hy~rochlor;~e
3-cyanobenzoic acid (5.88 g, 40 mmol) was suspended
in 50 mL THF and the mixture was warmed up with
stirring. After all solid went into solution, 50 mL
isopropanol was added and the solution was allowed to
cool to room temperature. To it was added 4.2 mL
precooled concentrated HCl followed by 300 mg
platinum(IV) oxide. The mixture was hydrogenated at 55
psi overnight. Ether (50 mL) was added, and the
precipitate was filtered, washed with ether and
dissolved in methanol. The catalyst was filtered off and
the solvent was removed under reduced pressure to give
6.2 g (82%) product. lH-NMR (DMSO-d6): ~=4.08 (d, 2H);
7.53 (t, lH); 7.80 (d, lH); 7.94 (d, lH); 8.10 (s, lH);
8.65 (s, 3H).
Step 7: Fm~c-T.-asp~rtyl(t-hutyl)-3-(~m;nomethyl)-
hen7O;c ~c;~
- To a solution of FmocAsp(But)OPfp (17.33 g, 30
mmol) and the compound of Step 6 (6.19 g, 33 mmol) in 50
mL DMF cooled in an ice bath was added 11.5 mL (66 mmol)
diisopropylethylamine, and after stirring at room
temperature for 5 h, 200 mL 5% citric acid was added and
-127-
WO94122910 PCT~S94/032~
~he solution wa~ extracted with ethyl acetate twice. The
combined extracts were washed with brine, dried (MgSO4),
and concentrated to give a solid which was washed with
ether-petroleum ether and dried. Yield 16.3 g (100%).
lH-NMR (DMSO-d6): ~=1.35 (s, 8H); 2.48 (dd, lH); 2.70
~dd, lH); 4.2-4.4 (m, 6H); 7.30 (t, 2H); 7.4-7.5 (m,
4H); 7.7-7.9 (m, 7H); 8.55 (t, lH); 12.92 (s, lH).
Step 8: Fmoc-T.-As~Artyl(t-hutyl)-3-
10 (~m; nomethyl)benzoyl-D-2-Am;nohutyryl-N~-methyl-NW Nw _
(bishenzyloxycArhonyl)-T.-~rginyl-glycine
A mixture containing the compound of Step 7 (10.89
g, 20 mmol), pentafluorophenol (4.05 g, 22 mmol) and DCC
(4.13 g, 20 mmol) in 50 mL THF was stirred at room
temperature overnight. Dicyclohexylurea was filtered
off, rinsed with THF, and the filtrate was concentrated.
To it was added a solution of the compound of Step 5
(14.25 g, 20 mmol) in 40 mL DMF followed by 7.32 mL (42
mmol) diisopropylethylamine. The mixture was stirred at
room temperature for 4 h, insoluble material was
filtered off, and the filtrate was added to 200 mL 3~
citric acid with stirring. The solution was extracted
with ethyl acetate twice and the combined extracts were
washed with brine, dried (MgSO4), and concentrated. The
residue was triturated with ether-petroleum ether to
give 22 g (98%) product. FAB-MS (MH+): Calculated
1125.5; Found 1125.7.
Step 9: CyclorL-As~axtyl(t-hutyl)-3-
(Am;nomethyl)benzoyl-D-2-aminobutyryl-N~
(h;shenzy1OxycArbonyl)-T-~rglnyl-glycyll
A solution of the compound of Step 8 (22.5 g, 20
mmol) and 4-dimethylaminopyridine (14.66 g, 120 mmol) in
100 mL DMF was stirred overnight at room temperature and
added slowly to a solution of TBTU (6.42 g, 20 mmol) in
200 mL DMF over 3 h and stirring was continued for 1 h.
-128-
W094/~910 PCT~S94/032~
21~07~
Ethyl acetate tlO00 mL) was added and the solution was
washed with 1% citric acid 2 times, brine 3 times and
concentrated to dryness. The residue was taken up in THF
and after filtration, the solvent was removed under
reduced pressure to give a solid which was washed with
ether and dried. Yield 16 g (90%). FAB-MS (MH+):
Calculated 885.4; Found 885.2.
Step 10: Cyclo r r~-~s~rtyl-3-(~m;nomethyl)henzoyl-D-2
~m;nohutyryl-Nw NW'-(b;shenzyloxyc~rhonyl)-T-~rg;nyl-
glycyll
A solution of the compound of Step 9 (16 g, 18
mmol) in 200 mL 50% TFA in methylene chloride was
stirred at room temperature for 1.5 h and then
concentrated. The residue was triturated with ether to
give 14.5 g (97%) product. FAB-MS (MH+): Calculated
829.4; Found 829.1.
Fx~m~le 301
Cyclo r r~-~spArtyllAcetoxymet~yl)-3-(~m;nomet~yl)h
D-2-~m;n~hutyryl-r~-~rg;~yl-glycyll
A mixture containing the compound of Step 10
(above) (1.42 g, 1.7 mmol), bromomethyl acetate (980 mL,
25 10 mmol) and triethylamine (976 mL, 7 mmol) in 10 mL DMF
was stirred at room temperature overnight. Ethyl acetate
was added and the solution was washed with brine 3
times, dried (MgSO4), concentrated, and dried. The
residue was taken up in 8 mL DMF and to it was added 130
30 mL (2 mmol) methanesulfonic acid followed by 150 mg 10%
palladium on carbon. The mixture was hydrogenated at
atmospheric pressure for 2 h, the catalyst was filtered
off, and the solution was diluted with water.
Purification using semipreparative HPLC gave 650 mg (51)
pure product. FAB-MS (MH+): Calculated 633.3; Found
633.2.
-129-
WO94/22910 PCT~S94/032~
?.J ~9~ --
E~ample 308
Cyclo r T~-~spartyl (~ivaloyloxymethyl)-3-
(aminomethyl)benzoyl-D-2-aminobutyryl-T.-arqinvl-glycyll
A mixture containing the compound of Step 10
(above) (4.14 g, 5 mmol), chloromethyl pivalate (4.3 mL,
30 mmol), triethylamine (2.8 mL, 20 mmol), NaI (4.5 g,
30 mmol) in 10 mL DMF was stirred at room temperature
for 18 h. Ethyl acetate (100 mL) was added and the
solution was washed with brine 3 times, dried (MgSO4),
and concentrated. The residue was taken up in 15 mL
ethyl acetate and passed through a silica gel column
using ethyl acetate-THF (1:1) as eluent to give 1.5 g
pure product. The product was dissolved in 10 mL DMF and
hydrogenated at atmospheric pressure using 10% palladium
on carbon (130 mg) in the presence of methanesulfonic
acid (100 mL) for 2 h. The catalyst was filtered off,
rinsed with DMF, and the solution was diluted with
water. Purification using semipreparative HPLC gave 1 g
(26%) pure product. FAB-MS (MH+): Calculated 675.3;
Found 675.3.
~.x~m~le 351
Cyclo r T.-as p~ rtyl-l;so~ro~yloxyc~rbonyl-oxymethyl)-3-
ml nomethyl) henzoyl-D-~-aminobutyryl-T-arginyl-glycyl 1
A mixture containing the compound of Step 10 (4.14
g, 5 mmol), chloromethyl isopropyl carbonate (4.58 g, 30
mmol), triethylamine (2.8 mL, 20 mmol), NaI (4.5 g, 30
mmol) in 10 mL DMF at stirred at room temperature for 18
h. Ethyl acetate (100 mL) was added and the solution was
washed with brine 3 times, dried (MgSO4), and
concentrated. The residue was taken up in 10 mL ethyl
acetate-THF (1:1) and passed through a silica column
using ethyl acetate-THF (1:1) as eluent to give 1.6 g
-130-
WO94/~910 PCT~S94/032~
2issor~
product. The product was dissolved in 10 mL DMF and
hydrogenated at atmospheric pressure using 10% palladium
on carbon (150 mg) in the presence of 130 mL for 2 h.
The catalyst was filtered off, rinsed with DMF, and the
solution was diluted with water. Purification using
semipreparative HPLC gave lg (25%) pure product. FAB-MS
(MH+): Calculated 667.3; Found 667.3.
Incorporated herein by reference in their entirety
are the following copending, commonly assigned U.S.
Patent Applications which are filed on the same day as
the present application: Attorney Docket No. DM-6535,
named inventors Maduskuie and Pesti; Attorney Docket No.
DM-6650, named inventors Zhang, Ma, and De Grado; and
Attorney Docket No. DM-6665, named inventors De Grado,
Dorow, Ward, and Xue.
Ut;llty
The compounds of this invention possess
antiplatelet efficacy, as evidenced by their activity in
standard platelet aggregation assays or platelet
fibrinogen binding assays, as described below. A
compound is considered to be active in these assays if
it has an ICso value of less than about 1 mM. Platelet
aggregation and fibrinogen binding assays which may used
to demonstrate the antiplatelet activity of the
compounds of the invention are described below.
Pl~telet Aggre~tion Ass~y: Venous blood was
obtained from the arm of a healthy human donor who was
drug-free and aspirin-free for at least two weeks prior
to blood collection. Blood was collected into 10 ml
citrated Vacutainer tubes. The blood was centrifuged
for 15 minutes at 150 x g at room temperature, and
-131-
WO94/22910 9~ ; PCT~S94/032
platelet-rich plasma (PRP) was removed. The remaining
blood was centrifuged for 15 minutes at 1500 x g at room
temperature, and platelet-poor plasma (PPP) was removed.
Samples were assayed on a aggregometer (PAP-4 Platelet
Aggregation Profiler), using PPP as the blank (100%
transmittance). 200 ~l of PRP was added to each micro
test tube, and transmittance was set to 0%. 20 ~l of
various agonists ~ADP, collagen, arachidonate,
epinephrine, thrombin) were added to each tube, and the
aggregation profiles were plotted (% transmittance
versus time). The results were expressed as %
inhibition of agonist-induced platelet aggregation. For
the ICso evaluation, the test compounds were added at
various concentrations prior to the activation of the
platelets.
Pl~telet-Fibr;noaen Rin~in~ Ass~y: Binding of
125I-fibrinogen to platelets was performed as described
by Bennett et al. ~1983) Proc. Natl. Acad. Sci. USA 80:
2417-2422, with some modifications as described below.
Human PRP (h-PRP) was applied to a Sepharose column for
the purification of platelet fractions. Aliquots of
platelets (5 X 108 cells) along with 1 mM calcium
chloride were added to removable 96 well plates prior to
the activation of the human gel purified platelets (h-
GPP). Activation of the human gel purified platelets
was achieved using ADP, collagen, arachidonate,
epinephrine, and/or thrombin in the presence of the
ligand, 125I-fibrinogen. The 125I-fibrinogen bound to
the activated, platelets was separated from the free
form by centrifugation and then counted on a gamma
counter. For an ICso evaluation, the test compounds
were added at various concentrations prior to the
activation of the platelets.
-132-
-
WO94/22910 21 S 9 ~ 7 ~ PCT~S94/032~
.
The novel cyclic glycoprotein IIb/IIIa compounds of
the invention also possess thrombolytic efficacy, that
is, they are capable of lysing (breaking up) already
formed platelet-rich fibrin blood clots, and thus are
useful in treating a thrombus formation, as evidenced by
their activity in the tests described below. Preferred
cyclic compounds of the present invention for use in
thrombolysis include those compounds having an ICso
value (that is, the molar concentration of the cyclic
compound capable of achieving 50% clot lysis) of less
than about 1 mM, more preferably an ICso value of less
than about 0.1 mM, even more preferably an ICso value of
less than about 0.01 mM, still more preferably an ICso
value of less than about 0.001 mM, and most preferably
an ICso value of about 0.0005 mM.
ICso determinations may be made using a standard
thrombolysis assay, as described below. Another class of
preferred thrombolytic compounds of the invention
include those compounds which have a Kd of C 100 nM,
preferably ~ 10 nM, most preferably 0.1 to 1.0 nM.
Thro~holyt-c A.~s~y: Venous blood was obtained
from the arm of a healthy human donor who was drug-free
and aspirin free for at least two weeks prior to blood
collection, and placed into 10 ml citrated Vacutainer
tubes. The blood was centrifuged for 15 minutes at 1500
x g at room temperature, and platelet rich plasma (PRP)
was removed. To the PRP was then added 1 x 10-3 M of
the agonist ADP, epinephrine, collagen, arachidonate,
serotonin or thrombin, or a mixture thereof, and the PRP
incubated for 30 minutes. The PRP was centrifuged for 12
minutes at 2500 x g at room temperature. The
supernatant was then poured off, and the platelets
remaining in the test tube were resuspended in platelet
poor plasma (PPP), which served as a plasminogen source.
-133-
W094/229l0 9~ PCT~S94/032~
.
The suspension was then assayed on a Coulter Counter
(Coulter Electronics, Inc., Hialeah, FL), to determine
the platelet count at the zero time point. After
obtaining the zero time point, test compounds were added
at various concentrations. Test samples were taken at
various time points and the platelets were counted using
the Coulter Counter. To determine the percent of lysis,
the platelet count at a time point subsequent to the
addition of the test compound was subtracted from the
platelet count at the zero time point, and the resulting
number divided by the platelet count at the zero time
point. Multiplying this result by 100 yielded the
percentage of clot lysis achieved by the test compound.
For the IC50 evaluation, the test compounds were added
at various concentrations, and the percentage of lysis
caused by the test compounds was calculated.
Platelet GrAnulAr Secretion Stu~ies. The role of
the claimed platelet GPIIb/IIIa receptor antagonists on
the modulation of platelet granular secretion from the
a-granules, dense granules or intracellular Ca+2 binding
proteins was e~Am;ned. This class of compounds did not
have any significant effect on platelet granular
secretion of plasminogen activator inhibitor type-l
(PAI-1) from a-granules, the mobilization of
intracellular calcium stores or the secretion of the
vasoconstrictor serotonin from the dense granules.
However, other antiplatelet agents such as aspirin or
the antithrombin hirudin has been shown to inhibit
platelet granular secretion of the antifibrinolytic
(PAI-1) or the vasoconstrictor (serotonin) . Hence the
combination between a universal antiaggregatory as well
as an inhibitor of platelet secretion might provide
optimal clinical benefits.
-134-
W094l22910 PCT~S94/032~
259~7~`
.
The novel cyclic compounds of the invention are
also useful in combination products, that is, in
pharmaceutical compositions containing the novel cyclic
compounds of the invention in combination with
anti-coagulant agents such as warfarin or heparin, or
antiplatelet agents such as aspirin, piroxicam or
ticlopidine, or thrombin inhibitors such as
boropeptides, hirudin or argatroban, or thrombolytic
agents such as tissue plasminogen activator,
anistreplase, urokinase or streptokinase, or
combinations thereof. Such combination products possess
anti-platelet and thrombolytic efficacy, as evidenced by
their activity in the tests described below.
These and other uses for the novel cyclic compounds
of this invention, and combination products containing
the same, will be readily apparent from the disclosures
herein.
Pl~telet GPIIh/IIIa Rin~;ng Affin;ty
In the human gel-purified platelet (h-GPP)
125I-fibrinogen binding assay, representative compounds
of the present invention demonstrated high affinity in
inhibiting the 125I-fibrinogen binding to h-GPP (ICso =
5- 100 nM) regardless of the agonist used. In an
enzyme-linked immunosorbent assay (ELISA) using purified
GPIIb/IIIa receptors obtained from human platelets, the
representative compounds of the invention demonstrated
direct inhibition of fibrinogen binding to RGD
recognition site(s), with an ICso f 0.5-10 nM. The
inhibitory efficacy of the presently claimed compounds
on fibrinogen binding to the platelet GPIIb/IIIa
receptor was shown to be related to the number of
binding sites, as is evident from the decrease in IC50
when platelet number was decreased.
-135-
WO94/22910 PCT~S94/032~
~9~ --
Compound A (Example 3) was shown to displace
125I-fibrinogen bound to activated platelets . In this
study, fibrinogen bound to activated platelets was
incubated for 20 minutes prior to the addition of
Compound A. This suggests a high affinity for Compound
A in displacing fibrinogen from an already formed
platelet-rich clot. This effect may explain the lytic
efficacy of the compounds of the present invention. A
high affinity binding (Kd = 0.1 nM) of 3H-labeled
Compound A to activated human platelets was determined
based on Scatchard analysis. Additionally, in the
purified GPIIb/IIIa-biotinylated fibrinogen ELISA,
Compound A demonstrated competitive inhibitory efficacy
with a Ki f 0.4 nM based on Michaelis-Menten analysis.
As shown below, in the human PRP aggregation assay,
Compound A was shown not only to inhibit platelet
aggregation induced by agonists, but also to deaggregate
platelets after the initiation of aggregation. The
deaggregation efficacy of Compound A was dependent on
its concentration and the time of addition
post-initiation of platelet activation. The earlier the
addition of Compound A after the induction of
aggregation, the greater its deaggregatory efficacy
(Fig. Ia).
The effect of Compound A on the lysis of a
pre-formed platelet-rich clot was also exAm;ned. In
this regard, the thrombolytic efficacy of Compound A was
also evaluat.ed (Fig. Ib and II). Compounds A and B
(Example 4) both demonstrated a significant lytic
efficacy of pre-formed platelet rich-clot (Fig. Ib).
Furthermore, Compound A demonstrated in vitro and in
vivo synergistic e`fficacy with standard thrombolytics in
lysing a platelet-rich thrombus (Fig. IIIb). A
concentration-dependent lytic effect with an ICso of
0.5-1.0 uM for compounds A and B was shown (Fig. Ib).
-136-
W094/~910 PCT~S94/032~
~ 21~
In contrast the tetrapeptide, RGDS, was shown to be
ineffective under similar conditions (Fig. Ib).
Additionally, in vitro studies revealed synergy
between Compound A (0.1-1.0 uM) and streptokinase,
urokinase or t-PA in lysing a pre-formed platelet-rich
clot (Fig. III). These results suggest an 7n vivo lytic
potential for disclosed compounds of the present
invention. Additionally, administration of these novel
antagonists is expected to significantly reduce the
dosage of a thrombolytic agent being used for clot lysis
and the prevention of reocclusion and/or restenosis. In
this regard, increasing evidence suggests that platelet
activation after thrombolytic therapy might have a
significant role in delaying reperfusion and abrupt
closure. Hence, the disclosed analogs might be an
effective adjunct to thrombolytic therapy or
angioplasty.
The cyclic glycoprotein IIb/III antagonist
compounds of this invention have also been shown to
displace 125I-fibrinogen bound to activated platelets in
a platelet-fibrinogen binding assay similar to the
platelet-fibrinogen binding assay previously described.
The results indicated that the compounds have a high
affinity in displacing fibrinogen from an already formed
platelet-rich clot. Although not intending to be bound
by any theory of operation, this result may help explain
the surprising thrombolytic efficacy possessed by
compounds of the invention, as illustrated in the
preceding examples.
Fiaure 1
Representative cyclic compounds of the present
invention, namely the compound of Example 3 (cyclo-(D-
Abu-NMeArg-Gly-Asp-Mamb; the compound of formula (II)
-137-
W094/229l0 PCT~S94/03223
whereln R1 and R2 are H, J is D-2-aminobutyric acid, K
is a-N-methylargininer L is glycine, and M is aspartic
acid) (designated here as Compound A) and the compound
of Example 4 (cyclo-(D-Val-NMeArg-Gly-Asp-Mamb; the
compound of formula (II) wherein R1 and R2 are H, J is
D-valine, K is a-N-methyl-arginine~ L is glycine, and M
is aspartic acid) (designated here as Compound B) were
then tested in the human PRP aggregation assay (Figure
Ia). Figure 1 shows the effect of 0.1 uM Compound on
the reversal of the aggregatory response (deaggregation)
to 10 uM ADP when added at 1.5 min post-initiation of
aggregation.
In the human PRP aggregation assay, representative
compounds of the present invention are shown not only to
inhibit platelet aggregation induced by agonists, but
also to deaggregate platelets after the initiation of
aggregation (Figure Ia). The deaggregation efficacy of
compound A was dependent on its concentration and the
time of addition post-initiation of platelet activation.
The earlier the addition of Compound A after the
induction of aggregation, the greater its deaggregatory
efficacy.
Compounds A and B were also tested at varying
concentrations using the thrombolytic assay described
above (Figure Ib). Figure Ib shows the lytic effect of
Compound A and B on an already formed platelet rich
clot. The clot was formed by incubating platelets with
a mixture of agonists (TEAC mixture), which consists of
thrombin (0.01 U/ml), epinephrine (250 uM), ADP (250
uM), and collagen (10 ug.ml), for 30 minutes. As a
comparison, a linear peptide of sequence
arginine-glycine-aspartic acid-serine (RGDS) was also
tested in the thrombolytic assay. The results are shown
in Figure I. The compounds of the invention (Compounds
A and B) demonstrated a significant effect on the lysis
of an already formed platelet-rich clot. As the results
-138-
WO94t22910 PCT~S94/032~
21~i907~
indicated, Compounds A and B had ICso values of about
0.5-1.0 uM. By comparison, the RGDS linear peptide was
much less effective, even at substantially higher
concentrations (ICso > 1 mM).
..
F;gure II
Compounds A and B was tested at a concentration of
0.001 mM using the thrombolytic assay described above,
with platelet stimulation being carried out using 1 x
10-3 M concentration of ADP. As a comparison, the
standard thrombolytics tissue plasminogen activator
(tPA; 10 ~g/ml), urokinase (UK; 900 units/ml) and
streptokina~e (SK; 500 units/ml) were also tested in the
thrombolytic assay. The results are shown in Figure II.
The compounds of the invention (Compounds A and B)
demonstrated a significant effect on the lysis of an
already formed platelet-rich clot, with Compound A
providing significantly better clot lysis than tissue
plasminogen activator, urokinase, and streptokinase, and
Compound B providing significantly better clot lysis
than streptokinase. As the results indicated, Compound
A had an excellent lysis percentage of 70% or greater.
F;~ure III
Figure III shows the effect of 1 uM of Compound A
on the lysis of an already formed platelet-rich clot.
The clot was formed by the addition of TEAC mixture
(which consists of thrombin (0.01 U/ml), epinephrine
(250 uM), ADP (250 uM), and collagen (10 ug.ml)) for 30
minutes. Compound A resulted in significant clot lysis
- 30 by itself as compared to tissue plasminogen activator
(tPA; 10 ~g/ml), urokinase (UK; 900 units/ml) and
streptokinase (SK; 500 units/ml). A synergy (greater
than additive effect between the standard thrombolytics
-139-
WO94/22910 PCT~S94/032~
~g~ "''' ~
and the IIb/IIIa antagonist Compound A was demonstrated.
Data represent mean + SEM, n = 3 in each group.
Figure V
Compound C was tested at a concentration of 1 uM
using the thrombolytic assay described above, both alone
and in combination with the standard thrombolytics
tissue plasminogen activator (tPA; 10 ~g/ml), urokinase
(UK; 900 units/ml) and streptokinase (SK; 500 units/ml).
As the results indicate, the combination of Compound C
with tissue plasminogen activator, urokinase or
streptokinase gave a greater than additive effect than
either agent alone.
Figure VI
Compound D was tested at a concentration of 1 uM
using the thrombolytic assay described above, both alone
and in combination with the standard thrombolytics
tissue plasminogen activator (tPA; 10 ~g/ml), urokinase
(UK; 900 units/ml) and streptokinase (SK; 500 units/ml).
As the results indicate, the combination of Compound D
with tissue plasminogen activator, urokinase or
streptokinase gave a greater than additive effect than
either agent alone.
Figure VII
VII a.
Effect of 1 uM Compound A on the lysis of an
already formed platelet-rich clot. The clot was formed
by the addtion of TEAC mixture (which consists of
thrombin [0.001 U/ml], epinephrine [250 uM], adenosine
diphosphate [250 uM] and collagen [10 ug/ml] for 30 min.
Compound A resulted in a significant clot lysis by
itself as compared to SK (500 U/ml), UK (900 U/ml) or
t-PA (10 ug/ml). A synergistic effect between the
-140-
WO94/22910 PCT~S94/032~
21~9~70
.
standard thrombolytics and the IIb/IIIa antagonist
Compound A was demonstrated. Data represent mean + SEM,
N=3 in each group.
VII b.
In vivo thrombolytic efficacy of Compound A and its
interaction with standard thrombolytics: Compound A at
1 mg/kg I.V. in the modified Lucchesi model resulted in
significant lysis of an already formed thrombus in the
femoral artery. Additionally, Compound A in combination
with sub-optimum doses of the standard thrombolytic,
streptokinase (75 KU) resulted in a significant
synergistic effect in fully lysing the thrombus with
subsequent restoration of flow, and the prevention of
reocclusion. Data represent mean + SEM. n=3-~ in each
group.
F;gure VIII
Effects of Compound A vs saline, when given to
anesthetized canine (male or female mongrel dogs) at 1.0
mg/kg I.V., on the incidence of femoral artery
reocclusion post-thrombolysis with streptokinase (250-
300 x 1000 IU/kg) or t-PA. Compound A resulted in 100%
prevention of reocclusion for a period > 240 minutes, in
comparison to saline-treated animals which were shown to
reocclude at 42 + 10 min. Compound A (1.0 mg/kg, I.V.)
resulted in % prevention of the incidence of reocclusion
post-thrombolysis with SK or t-PA. Data represents mean
+ SEM, n=6 in each group.
~nt;pl~telet co~h;n~t;on of the cycl;c GPIIh/III~
- receptor ~nt~on;st of the present ;nvention ~n~ ~sp;rin
~n~/or hep~r;n.
Methods: Twelve purpose bred mongrel dogs (8-15 months
of age) of either sex weighing between 8-12 kg were
-141-
WO94/22910 ~ PCT~S94/032~
2~9~rl ~
anesthetlzed with thiamylal sodium (15 mg/kg, l.v.) and
alpha-chloralose (100 ng/kg, i.v.) Dogs were placed on
positive pressure ventilation (15 mg/kg (a) 20
breaths/min). The femoral artery and vein were
dissected and cannulated for arterial blood pressure and
heart rate monitoring, blood sampling, and intravenous
injections.
Treatment Groups:
Group I (Saline): I.V. bolus of saline.
Group II (Aspirin): 10 mg/kg, po - 30 min prior to
blood sampling.
Group III (Compound A): 0.08 mg/kg, I.V. bolus.
Group IV (Aspirin/Compound A): Aspirin at 10 mg/kg,
po - 30 min prior to the administration of
Compound A at 0.08 mg/kg, i.v.
(a) Serial blood samples were withdrawn for ex
v vo platelet agregation and platelet counts.
(b) Bleeding time (min) was monitored over time as
well.
(c) Plasma levels of Compound A were determined by
an ELISA in all groups.
Results: The Compound A/aspirin, Compound A/heparin,
and Compound A/warfarin combinations demonstrated an
improved antiplatelet efficacy as compared to Compound A
alone. This was achieved without any significant
effects on bleeding time or platelet counts.
Dosage and Formulation
The compounds of this invention can be administered
by any means that produces contact of the active agent
with the agent's site of action, glycoprotein IIb/IIIa
(GPIIb/IIIa), in the body of a mammal. They can be
-142-
W094/22910 21~ 9 0 7 ~ PCT~S94/032~
.
administered by any conventional means available for use
in conjunction with pharmaceuticals, either as
individual therapeutic agents or in a combination of
therapeutic agents, such as a second antiplatelet agent
such as aspirin, piroxicam, or ticlopidine which are
agonist-specific, or an anti-coagulant such as warfarin
or heparin, or a thrombin inhibitor such as a
boropeptide, hirudin or argatroban, or a thrombolytic
agent such as tissue plasminogen activator,
anistreplase, urokinase or streptokinase, or
combinations thereof. The compounds of the invention,
or compounds of the invention in combination with other
therapeutic agents, can be administered alone, but
generally administered with a pharmaceutical carrier
selected on the basis of the chosen route of
administration and standard pharmaceutical practice.
The dosage of the novel cyclic compounds of this
invention ac~ministered will, of course, vary depending
upon known factors, such as the pharmacodynamic
characteristics of the particular agent and its mode and
route of administration; the age, health and weight of
the recipient; the nature and extent of the symptoms;
the kind of concurrent treatment; the frequency of
treatment; and the effect desired. A daily dosage of
active ingredient can be expected to be about 0.01 to 10
milligrams per kilogram of body weight.
Dosage forms (compositions suitable for
administration) contain from about 1 milligram to about
100 milligrams of active ingredient per unit. In these
pharmaceutic:al compositions the active ingredient will
ordinarily be present in an amount of about 0.5-95% by
weight based on the total weight of the composition.
The active ingredient can be administered orally in
solid dosage forms, such as capsules, tablets, and
powders, or in liquid dosage forms, such as elixirs,
-193-
WO94/22910 PCT~S94/03223
~ 9~ ~
syrups, and suspensions. It can also be administered
parenterally, in sterile liquid dosage forms.
Gelatin capsules contain the active ingredient and
powdered carriers, such as lactose, starch, cellulose
derivatives, magnesium stearate, stearic acid, and the
like. Similar diluents can be used to make compressed
tablets. Both tablets and capsules can be manufactured
as sustained release products to provide for continuous
release of medication over a period of hours. Compressed
tablets can be sugar coated or film coated to mask any
unpleasant taste and protect the tablet from the
atmosphere, or enteric coated for selective
disintegration in the gastrointestinal tract.
Liquid dosage forms for oral administration can
contain coloring and flavoring to increase patient
acceptance.
In general, water, a suitable oil, saline, aqueous
dextrose (glucose), and related sugar solutions and
glycols such as propylene glycol or polyethylene glycols
are suitable carriers for parenteral solutions.
Solutions for parenteral administration preferably
contain a water soluble salt of the active ingredient,
suitable stabilizing agents, and if necessary, buffer
substances. Antioxidizing agents such as sodium
bisulfite, sodium sulfite, or ascorbic acid, either
alone or combined, are suitable stabilizing agents.
Also used are citric acid and its salts and sodium EDTA.
In addition, parenteral solutions can contain
preservatives, such as benzalkonium chloride, methyl- or
propyl-paraben, and chlorobutanol.
Suitable pharmaceutical carriers are described in
Rem; ngton's PhAr-mAceutical Sciences, Mack Publishing
Company, a standard reference text in this field.
Useful pharmaceutical dosage-forms for
administration of the compounds of this invention can be
illustrated as follows:
-144-
W094/22910 PCT~S94/032~
~ 215907~
C~sules
A large number of unit capsules are prepared by
filling standard two-piece hard gelatin capsules each
with 100 milligrams of powdered active ingredient, 150
milligrams of lactose, 50 milligrams of cellulose, and 6
milligrams magnesium stearate.
Soft Gel~tin C~sules
A mixture of active ingredient in a digestable oil
such as soybean oil, cottonseed oil or olive oil is
prepared and injected by means of a positive
displacement pump into gelatin to form soft gelatin
capsules containing 100 milligrams of the active
ingredient. The capsules are washed and dried.
TAhlets
A large number of tablets are prepared by
conventional procedures so that the dosage unit was 100
milligrams of active ingredient, 0.2 milligrams of
colloidal silicon dioxide, 5 milligrams of magnesium
stearate, 275 milligrams of microcrystalline cellulose,
11 milligrams of starch and 98.8 milligrams of lactose.
Appropriate coatings may be applied to increase
palatability or delay absorption.
The combination products of this invention, such as
the novel cyclic IIb/IIIa antagonist compounds of this
invention in combination with an anti-coagulant agent
such as warfarin or heparin, or an anti-platelet agent
such as aspirin, piroxicam or ticlopidine, or a thrombin
- inhibitor such as a boropeptide, hirudin or argatroban,
or a thrombolytic agent such as tissue plasminogen
~ activator, anistreplase, urokinase or streptokinase, or
combinations thereof, can be in any dosage form, such as
-145-
W0.94l229l0 PCT~S94/03223
~90~ -
those described above, and can also be administered in
various ways, as described above.
In a preferred embodiment, the combination products
of the invention are formulated together, in a single
dosage form (that is, combined together in one capsule,
tablet, powder, or liquid, etc.). When the combination
products are not formulated together in a single dosage
form, the cyclic glycoprotein IIb/IIIa compounds of this
invention and the anti-coagulant agent, anti-platelet
agent, thrombin inhibitor, and/or thrombolytic agent may
be administered at the same time (that is, together), or
in any order, for example the compounds of this
invention are administered first, followed by
administration of the anti-coagulant agent,
anti-platelet agent, thrombin inhibitor, and/or
thrombolytic agent. When not administered at the same
time, preferably the administration of the compound of
this invention and any anti-coagulant agent,
anti-platelet agent, thrombin inhibitor, and/or
thrombolytic agent occurs less than about one hour
apart, more preferably less than about 30 minutes apart,
even more preferably less than about 15 minutes apart,
and most preferably less than about 5 minutes apart.
Preferably, administration of the combination products
of the invention is oral. The terms oral agent, oral
inhibitor, oral compound, or the like, as used herein,
denote compounds which may be orally administered.
Although it is preferable that the cyclic IIb/IIIa
antagonist compounds of this invention and the
anti-coagulant agent, anti-platelet agent, thrombin
inhibitor, and/or thrombolytic agent are both
administered in the same fashion (that is, for example,
both orally), if desired, they may each be administered
in different fashions (that is, for example, one
component of the combination product may be administered
orally, and another component may be administered
-146-
WO94/~910 PCT~S9q/032~
~ 2159~70
intravenously). The dosage of the combination products t
of the invention may vary depending upon various factors
such as the pharmacodynamic characteristics of the
particular agent and its mode and route of
5 administration, the age, health and weight of the
recipient, the nature and extent of the symptoms, the
kind of concurrent treatment, the frequency of
treatment, and the effect desired, as described above.
Although the proper dosage of the combination
10 products of this invention will be readily ascertainable
by one skilled in the art, once armed with the present
disclosure, by way of general guidance, where the cyclic
compounds of this invention are combined with
anti-coagulant agents, for example, typically a daily
dosage may be about 0.01 to 10 milligrams of the cyclic
compound of this invention and about l to 7.5 milligrams
of the anticoagulants, preferably about 0.1 to 1
milligrams of the cyclic compounds of this invention and
about 1 to 5 milligrams of the anti-coagulants, per
kilogram of patient body weight. With regard to a
typical dosage form of this type of combination product,
such as a tablet, the novel compounds of this invention
generally may be present in an amount of about 5 to 10
milligrams, and the anti-coagulants in an amount of
about l to 5 milligrams.
Where the novel compounds of this invention are
combined with another anti-platelet agent, by way of
general guidance, typically a daily dosage may be about
0.01 to 25 milligrams of the cyclic compounds of this
invention and about 50 to 150 milligrams of the
additional anti-platelet agents, preferably about 0.1 to
- 1 milligrams of the novel compounds of this invention
and about 1 to 3 milligrams of antiplatelet agents, per
kilogram of patient body weight. With regard to a
typical dosage form of this type of combination product,
such as a tablet, the novel compounds of this invention
-147-
W094/22910 ~gQ~ a PCT~S94103223
.
may be present, for example, in an amount of about 5
milligrams, and the additional anti-platelet agent in an
amount of about 150 milligrams, or, for example, in an
amount of about 25 milligrams of the cyclic compound of
this invention and about 50 milligrams of the additional
antiplatelet agent.
Further, in terms of general guidance, where the
novel compounds of this invention are combined with
thrombolytic agents, typically a daily dosage may be
about 0.1 to 1 milligrams of the cyclic compound of this
invention, per kilogram of patient body weight and, in
the case of the thrombolytic agents, the usual dosage of
the thrombolyic agent when administered alone may be
reduced by about 70-80% when administered with a
compound of the present invention. With regard to a
typical dosage form of this type of combination product,
such as a tablet, the novel compounds of this invention
may be present, for example, in an amount of about 10
milligrams.
As discussed above, where two or more of the
foregoing therapeutic agents are combined or
co-administered with the compounds of this invention,
generally the amount of each component in a typical
daily dosage and typical dosage form may be reduced
relative to the usual dosage of the agent when
administered alone, in view of the additive or
synergistic effect which would be obtained as a result
of addition of further agents in accordance with the
present invention.
Particularly when provided as a single dosage form,
the potential exists for a chemical interaction between
the combined active ingredients (for example, a novel
compound of this invention and an anti-coagulant such as
warfarin or heparin, or a novel compound of this
invention and an anti-platelet agent such as aspirin,
-148-
WO94122910 PCT~S94/032~
21~9~70
piroxicam or ticlopidine, or a novel compound of this
invention and a thrombin inhibitor such as a
boropeptide, hirudin or argatroban, or a novel compound
of this invention and a thrombolytic agent such as
tissue plasminogen activator, anistreplase, urokinase or
streptokinase, or combinations thereof). For this
reason, the preferred dosage forms of the combination
products of this invention are formulated such that
although the active ingredients are combined in a single
dosage form, the physical contact between the active
ingredients is minimized (that is, reduced).
In order to minimize contact, one embodiment of
this invention where the product is orally administered
provides for a combination product wherein one active
ingredient is enteric coated. By enteric coating one of
the active ingredients, it is possible not only to
minimize the contact between the combined active
ingredients, but also, it is possible to control the
release of one of these components in the
gastrointestinal tract such that one of these components
is not released in the stomach but rather is released in
the intestines. Another embodiment of this invention
where oral administration is desired provides for a
combination product wherein one of the active
ingredients is coated with a sustained-release material
which effects a sustained-release throughout the
gastrointestinal tract and also serves to minimize
physical contact between the combined active
ingredients. Furthermore, the sustained-released
component can be additionally enteric coated such that
the release of this component occurs only in the
intestine. Still another approach would involve the
formulation of a combination product in which the one
component is coated with a sustained and/or enteric
release polymer, and the other component is also coated
with a polymer such as a lowviscosity grade of
-149-
W094/22910 PCT~S94/03223
. .`, ~15q~
--
hy~roxypropyl methylcellulose (HPMC) or other
appropriate materials as known in the art, in order to
further separate the active components. The polymer
coating serves to form an additional barrier to
interaction with the other component.
Dosage forms of the combination products of the
present invention wherein one active ingredient is
enteric coated can be in the form of tablets such that
the enteric coated component and the other active
ingredient are blended together and then compressed into
a tablet or such that the enteric coated component is
compressed into one tablet layer and the other active
ingredient is compressed into an additional layer.
Optionally, in order to further separate the two layers,
one or more placebo layers may be present such that the
placebo layer is between the layers of active
ingredients. In addition, dosage forms of the present
invention can be in the form of capsules wherein one
active ingredient is compressed into a tablet or in the
form of a plurality of microtablets, particles, granules
or non-perils, which are then enteric coated. These
enteric coated microtablets, particles, granules or non-
perils are then placed into a capsule or compressed into
a capsule along with a granulation of the other active
ingredient.
These as well as other ways of minimizing contact
between the components of combination products of the
present invention, whether administered in a single
dosage form or administered in separate forms but at the
same time by the same manner, will be readily apparent
to those skilled in the art, once armed with the present
disclosure.
Pharmaceutical kits useful in, for example, the
inhibition of platelet aggregation, the treatment of
blood clots, and/or the treatment of thromboembolic
-150-
W094/229l0 PCT~S94/032
disorders, which comprise a therapeutically effective
amount of a novel cyclic platelet glycoprotein IIb/IIIa
compound of this invention along with a therapeutically
effective amount of an anti-coagulant agent such as
warfarin or heparin, or an antiplatelet agent such as
aspirin, piroxicam or ticlopidine, or a thrombin
inhibitor such as a boropeptide, hirudin or argatroban,
or a thrombolytic agent such as tissue plasminogen
activator, anistreplase, urokinase or streptokinase, or
combinations thereof, in one or more sterile containers,
are also within the ambit of the present invention.
Sterilization of the container may be carried out using
conventional sterilization methodology well known to
those skilled in the art. The sterile containers of
materials may comprise separate containers, or one or
more multi-part containers, as exemplified by the
UNIVIAL~ twopart container (available from Abbott Labs,
Chicago, Illinois), as desired. The novel compounds of
the invention and the anti-coagulant agent, anti-
platelet agent, thrombin inhibitor, thrombolytic agent,and/or combinations thereof, may be separate, or
combined into a single dosage form as described above.
Such kits may further include, if desired, one or more
of various conventional pharmaceutical kit components,
such as for example, one or more pharmaceutically
acceptable carriers, additional vials for mixing the
components, etc., as will be readily apparent to those
skilled in the art. Instructions, either as inserts or
as labels, indicating quantities of the components to be
administered, guidelines for administration, and/or
guidelines for mixing the components, may also be
included in the kit.
The Tables below set forth representative compounds
of the present invention. In the Tables below the
biological activity of the compounds is indicated as the
--1 5 1--
W094l22910 PCT~S94/03223
59~
ICso value in the platelet aggregation assay described
above. The ICso values are expressed as: +++ = ICso
value of less than 1 uM; ++ = ICso value of l uM to 10
uM; and ; + = ICso value of greater than 10 uM to about
100 uM. As used herein "uM" means micromolar. Where a
mixture of isomers of a compound were tested, for
example isomers designated as isomer 1 and isomer 2, the
biological activity of the mixture is indicated in
parentheses for each isomer.
-152-
PCT/US94/03223
WO 94/22910
~ gO7~
T~hl e
~ Gly~
NMeArg Asp
J\ NH
O=C ~ R40
l 11
-N-C-C-
~ ~ R10 J= Rl3 R5
The optical isomer of J is indicated. R10=H
unless otherwise indicated.
E~_ B3 B4 B5 FAB-MS O~t ica 1
NQ (M+H) I~m~r
1 H H H 533.26
2 H H CH3 547.23 (D)
2a H H CH3 (L)
3 H H CH2CH3 561.46 (D)
3a H H CH2CH3 (L)
3c H H CH2cH3 R10=I 687.33 (D)
4 H H CH(CH3)2 575-45 (D)
4a H H CH(CH3)2 (L)
5 H H CH2cH(cH3)2 589.48 (D)
5a H H CH2CH(CH3)2 (L)
6 H H CH2CH2CH3 ~D)
6a H H CH2CH2CH3 (L)
7 H H CH2CH2CH2CH3 589.26 (D)
7a H H CH2CH2CH2CH3 (L)
8 H H (CH2)sCH3 (D)
- 8a H H (CH2)5CH3 (L)
153
PCT~S94/03223
WO94/22910
~9~ --
T~hle 1 (con~inue~)
No. R4 R5 F~-MS o~t lcal IC50
9 H H (cH2)7CH3 (D)
9a H H (CH2)7CH3 (L)
10 H H C(CH3)3 (D)
lOa H H C(CH3)3 (L)
11 H H phenyl 609.27 (D)
lla H H phenyl (L)
12 H H phenylmethyl 623.28 (D)
12a H H phenylmethyl (L)
13 H H CH2OH (D)
13a H H CH2OH (L)
13b H H (cH2)3NH2 (D)
13c H H (cH2)3NH2 (L)
13d H H (CH2)3NHC(=NH)NH2 (D)
13e H H (CH2)3NHC(=NH)NH2 (L)
13f H H (cH2)4NH2 604.32 (D)
13g H H (cH2)4NH2 (L)
13h H H (CH2)4NHC(=NH)NH2 (D)
13i H H (CH2)4NHC(=NH)NH2 (L)
13j H H (cH2)5NH2 (D)
13k H H (cH2)5NH2 (L)
131 H H (CH2)sNHC(=NH)NH2 (D)
13m H H (CH2)sNHC(=NH)NH2 (L)
13n H H (CH2)4CH3 (D)
130 H H (CH2)4CH3 (L)
154
-
PCT/US94/03223
WO 94122910
~I590~
T~ h 1 e 1 ( cont; nue~l )
B3 B9 ~.5 FAR-MS O~tical ;E~50
- (M+H) I~
13p H H (cH2)6cH3 (D)
13q H H (cH2)6cH3 (L)
13r H H CH(CH3)CH2CH3 589.34 (D)
13s H H CH(CH3)CH2CH3 (L)
14 H H CH2SH (D)
14a H H CH2SH (L)
15 H H CH2OCH3 (D)
15a H H CH2OCH3 (L)
16 H H CH2SCH3 (D)
16a H H CH2SCH3 (L)
17 H H CH2CH2SCH3 (D)
17a H H CH2CH2SCH3 (L)
18 CH3 H H 547.34
19 H CH3 CH3
H CH2 CH2CH3
CH3
21 H H cyclopentyl (D)
21a H H cyclopentyl (L)
22 H H cyclohexyl (D)
22a H H cyclohexyl (L)
23 H H cyclohexylmethyl (D)
23c H H cyclohexylmethyl (L)
23a H H CH(CH3)2 R10=I 701.37 (D)
23b H H CH(CH3)2 R105I (L)
155
WO94/22910 PCT/US94/03223
~9~ --
23d H H H O
_(CH2)4 --N--C~N =N _N
23e H H H O o +++
--(CH2) ~ --N--C ~ C--O
23f H H H O NH2H +++
--(CH2 ) 4--N--C--C--C
23g H H H O
_(CH2)~ _N--C~
23h H H H O o +++
(cH2,~ I c ~a CH,
23j H H N(CH.)2 +++
H
--(CH2)4--N-- =o
23k H H H O H H o CH, + +
--(CH2)~--N_C--C--N--c--o--c--CH~
231 H H H O H H +++
-(CH2)~ - N- C -C-N -H
23m H H H O H H O o +++
--(CH2)~--N--C C--N--C ~ C~
H
23n H HH O +++
-(CH2)~ - N _ C
156
WO 94122910 PCT/US94/03223
90~
230 H H ¢~ +++
H o C=O
--(CH2)4 --N--C~
23p H H ~ ++
H O C=O
--(C~2)4 --N--
157
PCT/I~S94/03223
WO 94/22910
9~
T~hl e 1 (cont; nue~)
Gly ~
NMeArg Asp
J NH
O=C ~
Ex. J FAR--M.~ Opt; c~
No. (M+~:;) Isolner
24 ~> 573 . 46 (L)
~ 573 . 35 (D) +++
_N~
o
2 6 --NH~D_
o
2 7 -NH~
2 8 -NH ~Il~
< >< ( D )
N ~
158
WO94/~910 PCT~S94/032~
7 ~
T~hle 1
(cont.)
Ex. ~ FAR-M.S optlc~ so
~o. (M+S) Isomer
S (L)
28b < ~
o
-NHCH2CH2C(=O)- 547.28 +++
28c
~ (D)
28d
N
a
~ (L)
28e ~ >
N
11
Ex. B3 B4 B5 F~R-MS Optical I~so
No. (M+S) Iso~r
28f H H /==\639.54 (D) +++
- CH ~ OH
28g H H /==\(L)
- CH ~ OH
Specifically provided by the present invention are
those compounds of Table 1 wherein Asp is replaced by a
residue selected from: aMeAsp; ~MeAsp; NMeAsp; D-Asp;
Asp-(methylcarbonyloxymethyl ester)i
Asp-(ethylcarbonyloxymethyl ester);
Asp-(t-butylcarbonyloxymethyl ester);
Asp-(cyclohexylcarbonyloxymethyl ester)
159
-
PCT~S94/03223
WO94/22910
?~s9~Q
Asp~ (methylcarbonyloxy)ethyl ester);
Asp-(l-(ethylcarbonyloxy)ethyl ester);
Asp-(1-(t-butylcarbonyloxy)ethyl ester);
Asp-(l-(cyclohexylcarbonyloxy)ethyl ester);
Asp-(i-propyloxycarbonyloxymethyl ester);
Asp-(cyclohexylcarbonyloxymethyl ester);
Asp-(t-butyloxycarbonyloxymethyl ester);
Asp-(1-(i-propyloxycarbonyloxy)ethyl ester);
Asp-(1-(cyclohexyloxycarbonyloxy)ethyl ester);
Asp-(1-(t-butyloxycarbonyloxy)ethyl ester);
Asp-(dimethylaminoethyl ester);
Asp-(diethylaminoethyl ester);
Asp-((1,3-dioxa-5-methyl-cyclopenten-2-one-4-yl)methyl
ester);
Asp-((5-(t-butyl)-1,3-dioxa-cyclopenten-2-one-9-
yl)methyl ester);
Asp-((1,3-dioxa-5-phenyl-cyclopenten-2-one-4-yl)methyl
ester);
Asp-(1-(2-(2-methoxypropyl)carbonyloxy)ethyl ester).
160
PCT~S94/03223
WO94/22910
7 ~
T~hle 2
~ Gly~
Arg
J NH
o!c ~J R4 o
. 1 11
-N-C-C-
~ J H R5
The optical isomer of J is indicated.
Ex. g,4 E~5 FAR--M.S optical 1~50
No. (M+~) Is~m~r
29 H H 519.26 ++
H CH3 533.26 (D) ++
31 H CH3 533.25 (L)
32 H CH(CH3)2 561.22 (D) ++
32a H CH(CH3)2 (L)
33 H CH2CH(CH3)2 575.45 (D) ++
33a H CH2CH(CH3)2 (L)
34 H CH2CH3 547.21 (D) ++
34a H CH2CH3 (L)
35 H CH2OH 549.31 (D) ++
35a H CH2OH (L)
36 H phenylmethyl 609.25 (D) +
37 H phenylmethyl 609.26 (L) +
161
PCT~S94/03223
W094122910
~9~ --
T~hle 3
~ Gly~
K Asp
D-Val NH
O=\C ~
Il I
~ ~ = -N(R6)CH(R7)C(-o)-
The optical isomer of K is indicated.
Ex. ~6 ~7 FAR-M~ O~tical l~50
~Q (M+H) I~Q
32 H -(CH2)3NHC(=NH)(NH2) 561.22 (L)
32a H -(CH2)3NHC(=NH)(NH2) (D)
4 CH3 -(CH2)3NHC(=NH)(NH2) 575.45 (L)
4b CH3 -(CH2)3NHC(=NH)(NH2) 575.31 (D)
38 CH3 -(CH2)4NHC(=NH)(NH2) (L)
38a CH3 -(CH2)4NHC(=NH)(NH2) (D)
39 H -CH2 ~ CH2NH2 (L)
39a H -CH2 ~ CH2NH2 (D)
40 CH3 -CH2 ~ CH2NH2 595.23 (L)
4Oa CH3 -CH2 ~ CH2NH2 (D)
41 CH3 NH (L)
--CH2~CH2NH ~
NH2
162
PCT~S94/03223
WO94/22910
~ 21~9~7~
T~hle 3 (cont;nue~)
41a CH3 NH ~D)
- --CH2~cH2NH 4~
NH2
42 CH3 -CH2SCH2CH2NH2 (L)
42a CH3 -CH2SCH2CH2NH2 (D)
43 CH3 -CH2SCH2CH2NHC(=NH)(NH2) (L)
43a CH3 -CH2SCH2CH2NHC(=NH)(NH2) (D)
44 CH3 -CH2CH2SCH2CH2NH2 (L)
44a CH3 -CH2CH2SCH2CH2NH2 tD)
45 CH3 - (L)
CH2CH2SCH2CH2NHC(=NH)(NH
2)
45a CH3 - (D)
CH2CH2SCH2CH2NHC(=NH)(NH
2)
NH (L)
46 CH3
N~
NH (D)
46a CH
N~
/==\ ~NH (L)
47 CH3 - CH2 ~ C~NH2
/==\ ~NH (D)
47a CH3 - CH2 ~ C~NH2
~ (L)
48 CH3 -CH2 ~ cH2NH2
/~~\ (D)
48b CH3 -cH2 ~ cH2NH2
163
PCT/US94/03223
WO 94122910
~9~
T~hle 3 (cont; nue~
E~. ~6 B7 FAR--M.S Opt ica 1 ~5 o
~Q ~ ( M+H ) I ~ o~ r
4 8aCH3--CH2 ~NH-C~ (L)
48d CH3--CH2 ~NH-C~ (D)
A NH (L)
4 9CH 3 --CH2 ~ CH 2NH ~NH
49a CH3 ~CH2 {}CH2NH ~NH2 (D)
Ex .
No. ~ FAR--MS (M+H)
NH
52 lH ~NH2
>
_~ NH
164
PCT/US94/03223
WO 94/22910
2159~Q
TAhl e 4
~ L ~ A
NMeArg
D-Val NH
O=\C ~
F.PR-MS (M+H) ODti~A 1 ~Q
. Isorner
4 --NHCH2C (=O)-- 575 45 +++
54 -NHCH2CH2C (=O) - 58 9 . 32 ++
--OCH2C (=O)--
56 --OCH2CH2C (=O)--
57 -SCH2C (=O) -
58 -SCH2CH2C (=O)-
58c --NHCH (CH3) C (=O)-- 589 . 31 (L) +
165
WO94/22910 PCT~S94/03223
T~hle 5
~ Gly ~
NMeArg M
D-Val NH
O=C ~
~ M = -N(R10)C(R8)(R9)C(=o)-
R8 .B9 ~10 FAR--M.~ O~tical ,~50
No. tM+H) I~omf~r
4 -CH2COOH H H 575.45 (L) +++
63 -CH2COOH CH3 H 589.29 isomer 1 +t
63a -CH2COOH CH3 H 589.27 isomer 2 +
64 -CH(CH3)COOH H H 589.43 isomer 1 ++~
64a -CH(CH3)COOH H H 589.45 isomer 2 +
64b -CH2COOH H CH3 589.42 ++
64c -CH2COOH H H 575.42 (D) ++
66 -CH2SO3H H H
166
W094l22910 PCT~S94/03223
~159Q'7~1-
T~hle 6
~ Gly ~
NMeArg Asp
D-Val N-R2
. O--C, ~ Rl
I
~
The optical isomer of -CH(Rl)N(R2)- is indicated.
Ex. gl B2 E~= Opt;c~ 50
~Q. M.~(M+H) Isomer
4 H H 575.45 +++
68 CH3 H 589.31 isomer 1 +++
68a CH3 H 589.31 i~omer 2 +++
69 CH2CH3 H R
69a CH2CH3 H S
70 CH(CH3)2 H R
70a CH(CH3)2 H S
71 CH2CH2CH3 H R
7la CH2CH2CH3 H S
72 CH2CH2CH2CH3 H R
72a CH2CH2CH2CH3 H S
73 C(CH3)3 H R
73a C(CH3)3 H S
- 74 CH(CH3)CH2CH3 H R
74a CH(CH3)CH2CH3 H S
75 benzyl H R
75a benzyl H S
167
WO 94/22910 PCT/US94/03223
~9~ ~
T~hle 6
( cont inue~l )
~Im Bl B2 FAR- Opt ic~ 1
~1~ MS (M+H) Isomer
I~ .
76 phenyl H 651.33i~omer 1 ++
76a phenyl H 651.33i~omer 2 ++
77 cyclopentyl H R
77a cyclopentyl H S
78 cyclohexyl H R
78a cyclohexyl H S
79 H CH3 589.33
H CH2CH3
81 H CH2CH2CH3
82 H CH(CH3)2
83 H CH2cH2cH2cH3
84 H C(CH3)3
H CH (CH3) CH2CH3
86 H benzyl
168
W094122910 PCT~S94/03223
21~90~
T~hl e 7
F.x.No. Structure FAR-M.~ Q
IM+H~)
rMeAr9 ~ Gly
87 D-Val Asp 575 . 41 +++
0~
~NH
iqomer 1
rMeAr9 ~ Gly
88 ~ Asp 575. 44 +++
0~
I~,,NH
isomer 2
89a ~Gly~ 615.34 +++
NMeArg Asp
D-Va l NH
~J isomer 1
89b ~Gly~ 615.35 +++
NMeArg Asp
D-Va l NH
~J isomer 2
89c~GIy--ASp 625.32 ++
N MeAr~ \
D-Val NH
0~
169
WO 94/22910 PCT/US94/03223
9 T~hl e 8
Structure F~R~ o
E;~ r M+H 1
~Q-
Gly_
89d NMeArg A-p 687 . 33 +++
D-Abu NH
~'
ll
~1
89e ~ Gly_
dl-NMeOrn Al~p 533 . 34 ++
D-Abu N H
0~
b~l
N~MeAr~ ~ sp
1 589 +++
D-Val ~H
0~
N MQAr~ p
91 / 575 +++
D-Val ~H
~
170
WO 94/22910 PCT/US94/03223
~* 2~9~'~0
~Gly
N-MeAr~ p
92 1 575 +++
D-Val ~H
0 ~
171
PCT~S94/03223
W094/22910
, .: ~
- t
9~ ~ T~hle 9
~ - M
/ NR
o!c ~ CHR
R10 ~ R10a (VII)
wherein J = D-Val, X = NMeArg, ~ = Gly, M = Asp
F.xAm~le R10a ~10 FAR-M.~ Q
Nllmher (M+H)
93 Cl H 609 +++
94 I H 701.37 +++
MeO H 623(+H20)
96 Me H 589 +++
97 H Cl 609 +++
98 H I 701
99 H MeO 60S +++
100 H Me 589 +++
wherein J = D-Abu, R = NMeArg, L = Gly, ~ = Asp
100a H Cl 595.4 +++
100b H I 687.3 +++
100c H Me 575.4 +++
Specifically disclosed by the present invention are
those compounds of Tables 3-9 wherein D-Val is replaced by
a residue selected from: D-2-aminobutyric acid, D-Leu,
D-Ala, Gly, D-Pro, D-Ser, D-Lys, ~-Ala, Pro, Phe, NMeGly,
D-Nle, D-Phg, D-Ile, D-Phe, D-Tyr, Ala.
172
W094/22910 PCT~S94/03223
21~07~
T~hle lO
~ L ~ M
J/ NR2
O=C ~ CHR
~
wherein L = Gly, M = Asp, R2 and Rl = H,
K = -N(R6)CH(R7)C(=o)-
J = D-Val
Ex. _6 ~7 optic~ 50
No. Iso~er
101 CH3 ~ L
--(CH2)2~_~NH
102 CH3 ~ D
--(CH2)2~NH
103 CH3 L
--(CH2)3--CNH
104 CH3 ~ D
--(CH2)3~_~NH
105 CH3 ~ L
--(CH2)4--~_~NH
106 CH3 ~ D
--(CH2)4~NH
-173-
WO 94/22910 PCT/US94/03223
9~
107 CH3 ~--\ L
--CH20~ NH
108 CH3 /--\ D
--CH20~ NH
109 CH3 L
-CH20CH2--CNH
110 CH3 /--\ D
-CH20CH2 ~ _ ~NH
111 CH3 L
--t~H20(CH2)2{~NH
112 CH3 /--\ D
--~H20(CH2)2~_~NH
113 CH3 /--\ L
--(CH2)20~ NH
114 CH3 D
--(CH2)20--CNH
115 CH3 /--\ L
- (cH2)2ocH2~-~NH
116 CH3 /--\ D
- (CH2)20CH2~NH
--179--
WO 94/22910 PCT/US94/03223
~ 215!~Q7~
117 CH3 L
--CH2S {~NH
118 CH3 D
--CH2S{~NH
119 CH3 L
--CH2SCH2--~_ ~NH
120 CH3 /--\ D
--CH2SCH2~NH
121 CH3 L
--CH2S(CH2)2--CNH
122 CH3 D
--CH2S~CH2)2{~NH
123 CH3 /--\ L
--(CH2)2S--~ NH
124 CH3 D
--(CH2)2S--CNH
125 CH3 L
- (CH2)2SCH2--CNH
126 CH3 D
r\
- (CH2)2SCH2~NH
127 CH3--CH2--S--(cH2)3--NH--CH3 L
128 CH3--CH2--S--(cH2)3--NH--CH3 D
--175--
PCT~S94/03223
WO94122910
129 CH3 -CH2-S-(CH2)3-NH- L
CH2CH3
130 CH3 -CH2-S-(CH2)3-NH- D
CH2CH3
131 CH3 -CH2-S-(CH2)3-NH- L
CH(CH3)2
132 CH3 -CH2-S-(CH2)3-NH- D
CH(CH3)2
133 CH3 -CH2-S-(CH2)3-NH- L
CH2CH2--CH3
134 CH3 -CH2-S-(CH2)3-NH- D
CH2CH2-CH3
135 CH3 -CH2-S-(CH2)3-NH- L
C(CH3)3
136 CH3 -CH2-S-(CH2)3-NH- D
C(CH3)3
137 CH3 CH2-O-(CH2)3-NH-CH3 L
138 CH3 CH2-O-(CH2)3-NH-CH3 D
139 CH3 CH2-O-(CH2)3-NH- L
CH2CH3
140 CH3 CH2-O-(CH2)3-NH- D
CH2CH3
141 CH3 CH2-O-(CH2)3-NH- L
CH(CH3)2
142 CH3 CH2-O-(CH2)3-NH- D
CH(CH3)2
143 CH3 CH2-O-(CH2)3-NH- L
CH2CH2CH3
144 CH3 CH2-O-(CH2)3-NH- D
CH2CH2CH3
145 CH3 CH2-o-(cH2)3-NH- L
C(CH3)3
-176-
W094/22910 PCT~S94103223
~ ~lS~070
146 CH3 CH2-O-(CH2)3-NH- D
C(CH3)3
147 CH3 -CH2-S-(CH2)2-NH-CH3 L
148 CH3 -CH2-S-(CH2)2-NH-CH3 D
149 CH3 -cH2-s-(cH2)2-NH- L
CH2CH3
150 CH3 -CH2-S-(CH2)2-NH- D
CH2CH3
151 CH3 -CH2-S-(CH2)2-NH- L
CH(CH3)2
152 CH3 -cH2-s-(cH2)2-NH- D
CH(CH3)2
153 CH3 -CH2-S-(CH2)2-NH- L
CH2CH2--CH3
154 CH3 -CH2-S-(CH2)2-NH- D
~ CH2CH2--CH3
155 CH3 -cH2-s-(cH2)2-NH- L
C(CH3)3
156 CH3 -CH2-S-(CH2)2-NH- D
C(CH3)3
157 CH3 CH2-O-(CH2)2-NH-CH3 L
158 CH3 CH2-O-(CH2)2-NH-CH3 D
159 CH3 CH2-o-(cH2)2-NH- L
CH2CH3
160 CH3 CH2-O-(CH2)2-NH- D
CH2CH3
161 CH3 CH2-o-(cH2)2-NH- L
CH(CH3)2
162 CH3 CH2-O-(CH2)2-NH- D
CH(CH3)2
163 CH3 CH2-o-(cH2)2-NH- L
CH2CH2CH3
-177-
WO94/22910 PCT~S94103223
2~S9~
164 CH3 CH2-O-(CH2)2-NH- D
CH2CH2CH3
165 CH3 CH2-O-~CH2)2-NH- L
C(CH3)3
166 CH3 CH2-o-(cH2)2-NH- D
C(CH3)3
167 CH3 -CH2-S-(CH2)4-NH-CH3 L
168 CH3 -CH2-S-(CH2)4-NH-CH3 D
169 CH3 -CH2-S-(CH2)4-NH- L
CH2CH3
170 CH3 -CH2-S-~CH2)4-NH- D
CH2CH3
171 CH3 -CH2-S-(CH2)4-NH- L
CH(CH3)2
172 CH3 -CH2-S-(CH2)4-NH- D
CH(CH3)2
173 CH3 -CH2-S-(CH2)4-NH- L
CH2CH2-CH3
174 CH3 -CH2-S-(CH2)4-NH- D
CH2CH2-CH3
175 CH3 -CH2-S-(CH2)4-NH- L
C(CH3)3
176 CH3 -CH2-S-(CH2)4-NH- D
C(CH3)3
177 CH3 CH2-O-(CH2)4-NH-CH3 L
178 CH3 CH2-O-(CH2)4-NH-CH3 D
179 CH3 CH2-O-(CH2)4-NH- L
CH2CH3
180 CH3 CH2-O-(CH2)4-NH- D
CH2CH3
181 CH3 CH2-O-(CH2)4-NH- L
CH(CH3)2
-178-
W094/22910 PCT~S94/03223
2159 07D
.
182 CH3 CH2-O-~CH2)4-NH- D
CH(CH3)2
183 CH3 CH2-O-(CH2)4-NH- L
CH2CH2CH3
184 CH3 CH2-O-(CH2)4-NH- D
CH2CH2CH3
185 CH3 CH2-o-(cH2)4-NH- L
C(CH3)3
186 CH3 CH2-O-(CH2)4-NH- D
C(CH3)3
187 CH3 -CH2-S-(CH2)5-NH-CH3 L
188 CH3 -CH2-S-(CH2)5-NH-CH3 D
189 CH3 -CH2-S-(CH2)5-NH- L
CH2CH3
190 CH3 -CH2-S-(CH2)5-NH- D
CH2CH3
191 CH3 -CH2-S-(CH2)5-NH- L
CH(CH3)2
192 CH3 -CH2-S-(CH2)5-NH- D
CH(CH3)2
193 CH3 -CH2-S-(CH2)5-NH- L
CH2CH2--CH3
194 CH3 -CH2-S-(CH2)5-NH- D
CH2CH2-CH3
195 CH3 -CH2-S-(CH2)5-NH- L
C(CH3)3
196 CH3 -CH2-S-(CH2)5-NH- D
C(CH3)3
197 CH3 CH2-O-(CH2)5-NH-CH3 L
198 CH3 CH2-O-(CH2)5-NH-CH3 D
199 CH3 CH2-O-(CH2)5-NH- L
CH2CH3
-179-
WO94122910 ~5qQ~ PCT~S94/03223
20~ CH3 CH2-O-(CH2)5-NH- D
CH2CH3
201 CH3 CH2-o-(cH2)5-NH- L
CH(CH3)2
202 CH3 CH2-o-(cH2)5-NH- D
CH(CH3)2
203 CH3 CH2-O-(CH2)5-NH- L
CH2CH2CH3
204 CH3 CH2-O-(CH2)5-NH- D
CH2CH2CH3
205 CH3 CH2-O-(CH2)5-NH- L
C(CH3)3
206 CH3 CH2-O-(CH2)5-NH- D
C(CH3)3
207 CH3 -(cH2)2-s-(cH2)3-NH-cH3 L
208 CH3 -(CH2)2-S-(CH2)3-NH-CH3 D
209 CH3 -(cH2)2-s-(cH2)3-NH-cH2cH3 L
210 CH3 -(cH2)2-s-(cH2)3-NH-cH2cH3 D
211 CH3 -(cH2)2-s-(cH2)3-NH- L
CH(CH3)2
212 CH3 -(CH2)2-S-(CH2)3-NH- D
CH(CH3)2
213 CH3 -(CH2)2-S-(CH2)3-NH- L
CH2CH2--CH3
214 CH3 -(cH2)2-s-(cH2)3-NH- D
CH2CH2-CH3
215 CH3 -(CH2)2-S-(CH2)3-NH- L
C(CH3)3
216 CH3 -(cH2)2-s-(cH2)3-NH D
C(CH3)3
217 CH3 -(cH2)2-o-(cH2)3-NH-cH3 L
-180-
W094122910 PCT~S94/03223
~ 2lsso7a
218 CH3 -(cH2)2-o-(cH2)3-NH-cH3 D
219 CH3 -(CH2)2-O-(CH2)3-NH-CH2CH3 L
220 CH3 -~CH2)2-O-(CH2)3-NH-CH2CH3 D
221 CH3 -~CH2)2-O-(CH2)3-NH- L
CH(CH3)2
222 CH3 -(cH2)2-o-(cH2)3-NH- D
CH(CH3)2
223 CH3 -(CH2)2-O-(CH2)3-NH- L
CH2CH2CH3
224 CH3 -(CH2)2-O-(CH2)3-NH- D
CH2CH2CH3
225 CH3 -(CH2)2-O-(CH2)3-NH- L
C(CH3)3
226 CH3 -(cH2)2-o-(cH2)3-NH- D
C(CH3)3
227 CH3 -(cH2)2-s-(cH2)2-NH-cH3 L
228 CH3 -(CH2)2-S-(CH2)2-NH-CH3 D
229 CH3 -(CH2)2-S-(CH2)2-NH-CH2cH3 L
230 CH3 -(CH2)2-S-(CH2)2-NH-CH2CH3 D
231 CH3 -(cH2)2-s-(cH2)2-NH- L
CH(CH3)2
232 CH3 -(CH2)2-S-(CH2)2-NH- D
CH(CH3)2
233 CH3 -(cH2)2-s-(cH2)2-NH- L
CH2CH2--CH3
234 CH3 -(cH2)2-s-(cH2)2-NH- D
CH2CH2-CH3
235 CH3 -(cH2)2-s-(cH2)2-NH- L
C(CH3)3
236 CH3 -(CH2)2-S-(CH2)2-NH- D
C(CH3)3
-181-
PCT~S94/03223
WO94/22910
5~
237 CH3 -(cH2)2-o-(cH2)2-NH-cH3 L
238 CH3 -(cH2)2-o-(cH2)2-NH-cH3 D
239 CH3 -(CH2)2-O-(CH2)2-NH-CH2CH3 L
240 CH3 -(CH2)2-O-(CH2)2-NH-CH2CH3 D
241 CH3 -(CH2)2-O-(CH2)2-NH- L
CH(CH3)2
242 CH3 -(cH2)2-o-(cH2)2-NH- D
CH(CH3)2
243 CH3 -(CH2)2-O-(CH2)2-NH- L
CH2CH2CH3
244 CH3 -(cH2)2-o-(cH2)2-NH- D
CH2CH2CH3
245 CH3 -(cH2)2-o-(cH2)2-NH- L
C(CH3)3
246 CH3 -(CH2)2-O-(CH2)2-NH- D
C(CH3)3
247 CH3 -(CH2)2-S-(CH2)4-NH-CH3 L
248 CH3 -(CH2)2-S-(CH2)4-NH-CH3 D
249 CH3 -(CH2)2-S-(CH2)4-NH-CH2CH3 L
250 CH3 -(CH2)2-S-(CH2)4-NH-CH2CH3 D
251 CH3 -(CH2)2-S-(CH2)4-NH- L
CH(CH3)2
252 CH3 -(CH2)2-S-(CH2)4-NH- D
CH(CH3)2
251 CH3 -(CH2)2-S-(CH2)4-NH- L
CH2CH2-CH3
254 CH3 -(CH2)2-S-(CH2)4-NH- D
CH2CH2-CH3
255 CH3 -(CH2)2-S-(CH2)4-NH- L
C(CH3)3
-182-
PCT~S94/03223
WO94/~910
21~9070
.
256 CH3 -(cH2)2-s-(cH2)4-NH- D
C(CH3)3
257
CH3 -(CH2)3-NH-CH3 L
258 CH3 -(cH2)3-NH-cH3 D
259 CH3 -(CH2)3-NH-CH2CH3 L
260 CH3 -(cH2)3-NH-cH2cH3 D
261 CH3 -(CH2)3-NH-CH(CH3)2 L
262 CH3 -(cH2)3-NH-cH(cH3)2 D
263 CH3 -(CH2)3-NH-CH2CH2CH3 L
264 CH3 -(CH2)3-NH-CH2CH2CH3 D
265 CH3 -(cH2)3-NH-c(cH3)3 L
266 CH3 -(cH2)3-NH-c(cH3)3 D
267 CH3 -(CH2)4-NH-CH3 L
268 CH3 -(CH2)4-NH-CH3 D
269 CH3 -(CH2)4-NH-CH2CH3 L
270 CH3 -(CH2)4-NH-CH2CH3 D
271 CH3 -(CH2)4-NH-CH(cH3)2 L
272 CH3 -(CH2j4-NH-CH(CH3)2 D
273 CH3 -(CH2)4-NH-CH2CH2CH3 L
274 CH3 -(CH2)4-NH-CH2CH2CH3 D
275 CH3 -(cH2)4-NH-c(cH3)3 L
276 CH3 -(cH2)4-NH-c(cH3)3 D
277 CH3 -(CH2)5-NH-CH3 L
278 CH3 -(CH2)5-NH-CH3 D
279 CH3 -(cH2)5-NH-cH2cH3 L
280 C~3 -(cH2)5-NH-cH2cH3 D
-183-
-
WO 94122910 : : PCT/US94/03223
281 CH3 --(cH2)5-NH-cH(cH3)2 L
282 CH3 --(CH2)s--NH--CH(CH3)2 D
283 CH3 ~ (CH2)5--NH--CH2CH2cH3 L
284 CH3 --(cH2)s-NH-cH2cH2cH3 D
285 CH3 --(cH2)5-NH-c (CH3)3 L
286 CH3 ~ (cH2)5--NH--C (CH3)3 D
287 CH3 --(CH2) 6--NH--CH3 L
288 CH3 --(CH2) 6--NH--CH3 D
289 CH3 --(CH2) 6--NH--CH2CH3 L
290 CH3 --(CH2) 6-NH-cH2cH3 D
29l CH3 --(CH2) 6--NH--CH(CH3)2 L
292 CH3 ~ (CH2) 6--NH--CH(CH3)2 D
293 C~3 ~ (CH2) 6--NH--CH2CH2CH3 L
294 CH3 ~ (CH2) 6-NH-cH2cH2cH3 D
295 CH3 -(CH2) 6-NH-C (CH3)3 L
296 CH3 -(CH2) 6-NH-C (CH3)3 D
Specifically disclosed by the present invention are
those compounds of Table lO wherein D-Val is replaced by
a residue selected from: D-2-aminobutyric acid, D-Leu,
5 D-Ala, Gly, D-Pro, D-Ser, D-Lys, ~-Ala, Pro, Phe,
NMeGly, D-Nle, D-Phg, D-Ile, D-Phe, D-Tyr, Ala.
Also specifically disclosed by the present
invention are those compounds of Table lO wherein Asp is
10 replaced by a residue selected from: aMeAsp; ¦3MeAsp;
NMeAsp; D-Asp; Asp-(methylcarbonyloxymethyl ester);
Asp- (ethylcarbonyloxymethyl ester); .
Asp- (t-butylcarbonyloxymethyl ester);
Asp- (cyclohexylcarbonyloxymethyl ester);
--184--
WO94/22910 215 ~ 0 ~ 0 PCT~S94/03~3
Asp-(1-(methylcarbonyloxy)ethyl ester);
Asp-(1-(ethylcarbonyloxy)ethyl ester);
Asp-(1-(t-butylcarbonyloxy)ethyl ester);
Asp-(1-(cyclohexylcarbonyloxy)ethyl ester);
Asp-(i-propyloxycarbonyloxymethyl ester);
Asp-(cyclohexylcarbonyloxymethyl ester);
Asp-(t-butyloxycarbonyloxymethyl ester);
Asp-(l-(i-propyloxycarbonyloxy)ethyl ester);
Asp-(1-(cyclohexyloxycarbonyloxy)ethyl ester);
Asp-(1-(t-butyloxycarbonyloxy)ethyl ester);
Asp-(dimethylaminoethyl ester);
Asp-(diethylaminoethyl ester);
Asp-((1,3-dioxa-5-methyl-cyclopenten-2-one-4-yl)methyl
ester);
Asp-((5-(t-butyl)-1,3-dioxa-cyclopenten-2-one-4-
yl)methyl ester);
Asp-((1,3-dioxa-5-phenyl-cyclopenten-2-one-4-yl)methyl
ester);
Asp-(1-(2-(2-methoxypropyl)carbonyloxy)ethyl ester).
-185-
W094/22910 : ' PCT~S94/03223
T~hle 11
~Gly NH
NMeArg ~0
J = O R
NH
0~
W
J = D-Abu
Ex. B F~RM.
. (M+H)
301 -CH2-O-C(=O)-CH3 633.2
302 -CH2-O-C(=O)-CH2CH3
303 -CH2-O-C(=O)-CH(CH3)2
304 -CH2-O-C(=O)-(CH2)2-CH3
305 -CH2-O-C(=O)-(CH2)3-CH3
306 -CH2-O-C(=O)-CH2-CH(CH3)2
307 -cH2-o-c(=o)-cH(cH3)-cH2-cH3
308 -CH2-O-C(=O)-C(CH3)3 675.3
309 -CH2-O-C(=O)-cyclopropyl
310 -CH2-O-C(=O)-cyclobutyl
311 -CH2-O-C(=O)-cyclopentyl
312 -CH2-O-C(=O)-cyclohexyl
313 -CH2-O-C(=O)-phenyl
314 -CH2-O-C(=O)-4-methylphenyl
315 -CH2-O-C(=O)-4-ethylphenyl
-186-
WO94122910 2 ~ 5 9 0 7 o PCT~S94/03223
316 -CH2-O-C(=O)-4-isopropylphenyl
317 -CH2-O-C(=O)-4-propylphenyl
318 -CH2-O-C(=O)-4-t-butylphenyl
319 -CH2-O-C(=O)-4-methoxyphenyl
320 -CH2-O-C(=O)-4-ethoxyphenyl
321 -CH2-O-C(=O)-4-isopropyloxyphenyl
322 -CH2-O-C(=0)-4-propyloxyphenyl
323 -CH2-O-C(=O)-4-~-butoxyphenyl
324 -CH2-O-C(=O)-4-biphenyl
325 -CH(CH3)-O-C(zO)-CH3
326 -CH(CH3)-O-C(=O)-CH2CH3
327 -CH(CH3)-O-C(=O)-CH(cH3)2
328 -cH(cH3)-o-c(=o)-(cH2)2-cH3
329 -CH(CH3)-O-C(-O)-(CH2)3-cH3
330 -CH(CH3)-O-C(=O)-CH2-CH(CH3)2
331 -CH(CH3)-O-C(=O)-CH(CH3)-CH2-CH3
332 -CH(CH3)-O-C(=O)-C.(CH3)3
333 -CH(CH3)-O-C(=O)-cyclopropyl
334 -CH(CH3)-O-C(=O)-cyclobutyl
335 -CH(CH3)-O-C(=O)-cyclopentyl
336 -CH(CH3)-O-C(=O)-cyclohexyl
337 -CH(CH3)-O-C(=O)-phenyl
338 -CH(CH3)-O-C(=O)-4-methylphenyl
339 -CH(CH3)-O-C(=O)-4-ethylphenyl
340 -CH(CH3)-O-C(=O)-4-isopropylphenyl
341 -CH(CH3)-O-C(=O)-4-propylphenyl
-187-
PCT~S94/03223
W094122910
342 -CH(CH3)-0-C(=0)-4-~-butylphenyl
343 -CH(CH3)-0-C(=0)-4-methoxyphenyl
344 -CH(CH3)-0-C(=0)-4-ethoxyphenyl
345 -CH(CH3)-0-C(=0)-4-
isopropyloxyphenyl
346 -CH(CH3)-0-C(=0)-4-propyloxyphenyl
347 -CH(CH3)-0-C(=0)-4-~-butoxyphenyl
348 -CH(CH3)-0-C(=0)-4-biphenyl
349 -CH2-0-C(=0)-0-CH3
350 -CH2-0-C(=0)-0-CH2CH3
351 -CH2-0-C(=0)-0-CH(cH3)2 667.3
352 -CH2-0-C(=0)-0-(CH2)2-CH3
353 -CH2-0-C(=0)-0-(CH2)3-CH3
354 -cH2-o-c(-o)-o-cH2-cH(cH3)2
355 -CH2-0-C(=0)-0-CH(CH3)-CH2-CH3
356 -CH2-G-C(=0)-0-C(CH3)3
357 -CH2-0-C(=0)-0-cyclopropyl
358 -CH2-0-C(=0)-0-cyclobutyl
359 -CH2-0-C(=0)-0-cyclopentyl
360 -CH2-0-C(=0)-0-cyclohexyl
361 -CH2-0-C(=0)-0-phenyl
362 -CH2-0-C(=0)-0-4-methylphenyl
363 -CH2-G-C(=0)-0-4-ethylphenyl
364 -CH2-0-C(=0)-0-4-isopropylphenyl
365 -CH2-0-C(=0)-0-4-propylphenyl
366 -CH2-0-C(=0)-0-4-~-butylphenyl
-188-
PCT/US94/03223
W094/229l0
21~907û
367 -CH2-0-C(=0)-0-4-methoxyphenyl
368 -CH2-0-C(=0)-0-4-ethoxyphenyl
369 -CH2-0-C(=0)-0-4-
isopropyloxyphenyl
370 -CH2-0-C(=0)-0-4-propyloxyphenyl
371 -CH2-0-C(=0)-0-4-~-butoxyphenyl
372 -CH2-0-C(=0)-0-4-biphenyl
373 -CH(CH3)-0-C(=0)-0-CH3
374 -CH(CH3)-0-C(=0)-0-CH2CH3
375 -CH(CH3)-0-C(=0)-0-CH(CH3)2
376 -cH(cH3)-o-c(=o)-o-(cH2)2-cH3
377 -CH(CH3)-0-C(=0)-0-(CH2)3-cH3
378 -CH(CH3)-0-C(=0)-0-CH2-CH(CH3)2
379 -CH(CH3)-0-C(=0)-0-CH(CH3)-CH2-CH3
380 -CH(CH3)-0-C(=0)-0-C(CH3)3
381 -CH(CH3)-0-C(=0)-0-cyclopentyl
382 -CH(CH3)-0-C(=0)-0-cyclobutyl
383 -CH(CH3)-0-C(=0)-0-cyclopentyl
384 -CH(CH3)-0-C(=0)-0-cyclohexyl
385 -CH(CH3)-0-C(=0)-0-phenyl
386 -CH(CH3)-0-C(=0)-0-4-methylphenyl
387 -CH(CH3)-0-C(=0)-0-4-ethylphenyl
388 -CH(CH3)-0-C(-0)-0-4-
isopropylphenyl
389 -CH(CH3)-0-C(=0)-0-4-propylphenyl
390 -CH(CH3)-0-C(=0)-0-4-t-butylphenyl
-189-
W094l22910 PCT~S94/03223
9~
391 -CH(CH3)-0-C(=0)-0-4-methoxyphenyl
392 -CH(CH3)-0-C(=0)-0-4-ethoxyphenyl
393 -CH(CH3)-0-C(=0)-0-4-
isopropyloxyphenyl
394 -CH(CH3)-0-C(=0)-0-4-
propyloxyphenyl
395 -CH(CH3)-0-C(=0)-0-4-~-
butoxyphenyl
396 -CH(CH3)-0-C(=0)-0-4-biphenyl
397 CH2-N(CH3)2
398 CH2-N(CH2-CH3)2
399 CH2CH2-N(CH3)2
400 CH2-CH2-N(CH2CH3)2
401 CH2-CH2'
402 ~
CH2-CH2-NJ
403 CH2-cH2-N ~
404 -cH(cH3)oc(=o)c(cH3)2ocH3
405 0
0~0
~ CH3
406 0
~0
tBu
--1 9 0--
W094/22910 PCT~S94/03223
215907D
407 O
0~0
phenyl
408 CH2C(=O)OCH3
409 CH2C(=~)O-tBu
Specifically disclosed by the present invention are
those compounds of Table 10 wherein D-Abu is replaced by
a residue selected from: D-Val, D-Leu, D-Ala, Gly,
D-Pro, D-norvaline, D-Ser, D-Lys, ~-Ala, Pro, Phe,
NMeGly, D-Nle, D-Phg, D-Ile, D-Phe, D-Tyr, Ala.
--191--
