NZ263456A - Cyclic compounds useful as inhibitors of platelet glycoprotein iib/iiia and medicaments thereof - Google Patents

Description

New Zealand No. 263456 International No. PCT/US94/03223 Priority Date(s): Complete Specification Fifed: ttess. (6) coi.»<.Stj.o.^pa,.ia;.

...Bb\.&3&|.Q5rcM£t Publication Date:.. i P.O. Journal No: NEW ZEALAND PATENTS ACT 1953 COMPLETE SPECIFICATION Title of Invention: Cyclic compounds useful as inhibitors of platelet glycoprotein llb/llla Name, address and nationality of applicant(s) as in international application form: THE DU PONT MERCK PHARMACEUTICAL COMPANY, 1007 Market Street, Wilmington, Delaware 19898, United States of America us c©»v^eu-v>^-~ / 263456 TITLE Cyclic Compounds Useful as Inhibitors of Platelet 5 Glycoprotein Ilb/IIIa FTFT.n n,T THF. TWVFMTTnW This invention relates to novel cyclic compounds 10 containing carbocyclic ring systems useful as antagonists of the platelet glycoprotein Ilb/IIIa complex, and to pharmaceutical compositions containing such cyclic ccntpounds, with or without other therapeutic agents, for the inhibition of platelet aggregation, as throntoolytics, and/or for the 15 treatment of other thronboenbolic disorders.

BftCKfiROTTND OF THE INVENTION Activation of platelets and the resulting platelet aggregation and secretion of factors by the platelets has oeen 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, stroke, atherosclerosis and diabetes. The contribution of platelets to these disease processes 30 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 3 5 arterial thrombosis. Platelet activation has been shown to be enhanced during coronary thrombolysis which can 263456 2 0 lead to delayed reperfusion and reocclusion. Clinical studies with aspirin, ticlopidine and a monoclonal antibody for platelet glycoprotein Ilb/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 Ilb/IIIa complex (GPIIb/IIIa), which is the membrane protein mediating platelet aggregation. A recent review of GPIIb/IIIa is provided by Phillips et al. (1991) 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 I 1 - OCT 1996 26115, 6 antibody for GPIIb/IIIa indicate the benefit of a GPIIb/IIIa antagonist.

There is presently a need for a GPIIb/IIIa-specific antiplatelet agent which inhibits the activation and 5 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 10 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 15 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 20 binding and prevent the formation of platelet thrombi. For example, see Cadroy et al. (1989) J. Clin. Invest. 84: 939-9441; Klein et al.U.S. Patent 4,952,562, issued 6/28/90; European Patent Application EP 0319506 A; European Patent Application EP 0422 938 Al; European 25 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 91/0424 7; and European Patent Application EP 0343085 Al.

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.

SUMMARY OF THE INVENTION r*. c, WENT GfRCfcl ........Z] 263456 This invention provides novel cyclic compounds containing carbocyclic ring systems useful as antagonists of the platelet glycoprotein Ilb/IIIa . complex, and pharmaceutical compositions containing such cyclic 5 conpourds, for the inhibition of platelet aggregation, as thrcnbolytica, and/or for the treatment of thronixenbolic disorders. deaggregatory and thrombolytic effects of the cyclic Ilb/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 30 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 IC50 of about 0.0005 mM for Compound A. By comparison, the BRTF.F nr.SrRTPTTON OF THE FTGHREfi Fipurp T. Figure 1 shows the platelet N.2. PATENT OfFlCE 1- OCT 1996 linear peptide RGDS was much less effect thrombolytic, even at substantially higher concentrations.

Fiffurg ii. Figure II shows the thrombolytic effect of the cyclic Ilb/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), 10 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 15 plasminogen activator, urokinase, and streptokinase. effect of the cyclic compound cyclo-(D-AbuNMeArg-Gly-Asp-Mamb) (Compound A) and the standard thrombolytics 20 tissue plasminogen activator (tPA), urokinase (UK), and streptokinase (SK), both alone and in combination, on an already formed platelet-rich clot. The clot was formed by incubating the platelets with agonists for 30 minutes. Compound A showed a significant thrombolytic 25 effect, providing significant clot lysis at 1.0 uM. Moreover, Compound A in combination with tissue plasminogen activator, urokinase, or streptokinase was significantly better than Compound A alone, and significantly better than the additive effects of both 30 agents administered alone. of the cyclic Ilb/IIIa antagonist compound cyclo-(D-Val-NMeArg-Gly-Asp-Mamb) (Compound B) and the standard 35 thrombolytics tissue plasminogen activator (tPA), urokinase (UK) and streptokinase (SK), both alone and in Figure III shows the thrombolytic Figure IV shows the thrombolytic effect WO 94/22910 PCT/US94/03223 263456 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 5 better clot l-ysis 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 10 effects of both agents.

Figure 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 15 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. 2C Compound C ^lone showed a significant thrombolytic effect. It. corrJcination 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. ri.amre 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 30 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 35 effect. In combination with tissue plasminogen activator, urokinase or streptokinase, a thrombolytic -6- :.;Zi 2634 effect was achieved which was greater than the additive effect of the agents when administered alone. fiyurp vit. Figure VII shows the in vivo 5 thrombolytic and anti-thrombotic effect of the cyclic glycoprotein Ilb/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 10 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 15 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.

Figure vitt. 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 25 to reocclusion and percentage of reocclusion. The saline control showed 100% reocclusion, whereas administration of Compound A resulted in virtually no reocclusion.

This invention is directed to novel compounds of the formula (I): DETAILED DESCRIPTION OF THE INVENTTOW PATENT OfFlCC 1 - OCT 1996 263 4 Y: NR2 / \ (CHR1)„ R»0 or a pharmaceutically acceptable salt form thereof wherein: R1 is H, C1-C4 alkyl, phenyl, benzyl or phenyl (C1-C4) alkyl; p.2 is H or methyl; R10 is H, halogen, Ci-Cg alkyl, phenyl or C1-C4 alkoxy; n is 0-2; n' is 0-1; J is D-Ala, D-Val, D-Ile, D—Leu, D-Nle, D-phenylGly, DHPhe, D-D-Orn, D-Met, D-Pro, p-Ala, D-Tyr, D-Ser, NMeGly, D-cyclohexylGly, D-cyclohexylitiethylGly, D-norvaline, D-2-aminobutyric acid, D-2-aminopentanoic acid, Gly, N®-f azidobenzoyl-D-Lys, Nc-p-benzoylbenzoyl-D-Lys, I^-tryptophanyl-D-Lys, benzylbenzoyl-D-Lys, N^-p-acetobenzoyl-D-Lys, Ne-dansyl-D-Lys, Nc~t-butoxycarbonylglycyl-D-Lys, Nc-glycyl-D-Lys, Ne-p-benzoylbenzoylglycy1-D-Lys, Ne-p-phenylbenzoyl-D-Lys N^-iri-benzoylbenzoyl-D-Lys, or t^-o-benzoylbenzoyl-D-Lys; N.Z. PATENT -OFFICE 1- OCT 1996 263456 is OtNMeArg, N^N^diMe-N^-guanidinylOrn, N^MeLys or Ne, Na-diMeLys / is Gly; is selected from the group consisting of: Asp-(methylcarbonyloxymethyl ester); Asp-(ethylcarbonyloxymethyl ester); Asp-(t-butylcarbonyloxymethyl ester)/ Asp-(cyclohexylcarbonyloxymethyi 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-(cyclohexylcarbonyloxymethyi 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)-l,3-dioxa-cyclopenten-2-one-4-yl)methyl ester); Asp-((l,3-dioxa-5-phenyl-cyclopenten-2-one-4-yl)methyl ester); and Asp-(1-(2-(2-methoxypropyl)carbonyloxy)ethyl ester)- Ihis invention includes compounds of the formula (II): 263456 o / ,CHR1 ) n R10 (II) or a pharmaceutical^ acceptable salt form thereof wherein: R1 is H, C1-C4 alkyl, phenyl, benzyl or phenyl(C1—C4)alky1; R2 is H or methyl; R10 is H, halogen, Ci-Cg alkyl, phenyl or C1-C4 alkoxy; alternatively, when R10 is para to the carbonyl, R10 and R1 may be taken together to form -CH2-CH2-CH2- thereby to form a six-membered fused ring; n is 0—1; J is D-Ala, D-Val, D-Ile, D-Leu, D-Nle* D-phenylGly, D-Phe, D-Lys D-Orn, D-Met, D-Pro, p-Ala, D-Tyr, D-Ser, NMeGly, D-cyclohexylGly, D-cyclohexylmethylGly, D-norvaline, D-2-aminobutyric acid, D-2-aminopentanoic acid, Ne-p-a z idobenzoy1-D-Lys, Ne-p-benzoylbenzoy1-D-Lys, N€-tryptophanyl-D-Lys, Nc-o-ben.zylbenzoyl-D-Lys, Nc-p—acetobenzoyl-D—Lys, Ne-dansyl-D-Lys, Ne-t-butoxycarbonylglycyl-D-Lys, Ne-glycyl-D-Lys, Nc-p-benzoylbenzoylglycyl-D-Lys, Ne-p-phenylbenzoyl-DrLys, Ne-m-benzoylbenzoy1-D-Lyg, or N6-o-benzoylbenzoyl-D-Lys; K is CtNMfjArg, N^, N^diMe-N^guanidinylOrn, Na-MeLys or N£, Na-diMeLys; 26345 L is Gly; and M is selected from the group consisting of: Asp—(methylcarbonyloxymethyl ester); Asp—(ethylcarbonyloxymethyl ester); Asp-(t-butylcarbonyloxymethyl ester); Asp-(cyclohexylcarbonyloxymethyi ester); Asp-(1-(methylcarbonyloxy)ethyl ester); Asp-(1-(ethylcarbonyloxy)ethyl ester); Asp-(1-(t-butylcarbonyloxy)ethyl ester); Asp-(1-(cyclohexvlcarbonyloxy)ethyl ester); Asp— (i-propyloxycarbonyloxyitiethy 1 ester); Asp-(cyclohexylcarbonyloxymethyi 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-y1)methyl ester); and Asp-(1-(2-(2-methoxypropyl)carbonyloxy)ethyl ester).

The present invention includes compounds of formula (XI) above, or phaimaceutically acceptable salts thereof, wherein: R1 is H, Ci~C4 alkyl, phenyl, benzyl or phenyl(C1-C4) alkyl; R2 is H or methyl; R10 is H, halogen, Ci-Cs alkyl/. phenyl or C3.-C4 alkoxy; RTPATENT OfTICE 1 - OCT ©86 263 456 alternatively, when R10 is para to the caxbonyl, R10 and R1 may be taken together to form -CH2-CH2-CH2- thereby to form a six-membered fused ring; J is D-Ala, D-Val, D-Ile, D-Leu, B-Nle, D-phenylGly, D-Phe, D-Lys# D-Qrn, D-Met, D-Pro, Jl-Ala, D-Tyr, D-Ser, NMeGly, D-cyclohexylGly, D-cyclohexylmethylGly, D-norvaline/ D-2-aminobutyric acid, Ne-p-azidobenzoy1-D-Lys, Ne-p-benzoylbenzoyl-D-Lys, N®-tryptophany1-D-Ly3, Ne-o-ben.zylbenzoyl-D— Lys, Ne-p-acet,obenzoyl-D-Lys, Nc-dansyl-D-Lys, Ne-t-butoxycarbonylglycyl-D-Lys, Ne-glycyl-D-Lys, Ne-p-benzoylbenzoylglycyl-D-Lys, Ne-p-phenylbenzoy 1-D-Lys, Nc-JJi-benzoylbenzoyl-D-Lys, or Ne-o-benzoylbenzoyl-D-Lys; K is ocNMeArg; L is Gly; and M is selected from the group consisting of: Asp-(methylcarbonyloxymethy ester); Asp-(ethylcarbonyloxymethyl ester); Asp-(t-butylcarbonyloxymethyl ester); Asp-(cyclohexylcarbonyloxymethyi 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-(cyclohexylcarbonyloxymethyi 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); n is 1; 263456 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); and Asp-(1-(2-(2-methoxypropyl)carbonyloxy)ethyl ester), This invention includes ociqpounds of formula (IX), or pharmaceutically' acceptable salts thereof, wherein: R1 and R2 are independently selected from H or methyl; R10 is H; alternatively, when R10 is para to the carbonyl, R10 and. Rl may be taken together to form -CH2-CH2--CH2- thereby to form a six-membered fused ring; J is selected from: D-Val, D-2-aminobutyric acid, D-Leu, D-Ala, D-Pro, D-Ser, D-Lys, J3-Ala, NMeGly, D-Nle, D-phenylGly, D-Ile, D-Phe, or D-Tyr; K is CtNMeArg; L is Gly; m is selected from the group consisting of: Asp-(methylcarbonyloxymethj ester); Asp-(ethylcarbonyloxymethyl ester); Asp-(fc-butylcarbonyloxymethyl ester); Asp-(cyclohexylcarbonyloxymethyi ester); Asp-(1-(methylcarbonyloxy) ethyl ester); Asp-(1-(ethylcarbonyloxy)ethyl ester); Asp-(1-(t-butylcarbonyloxy)ethyl ester); Asp-(1-(cyclohexylcarbonyloxy)ethyl ester); 263456 Asp-(i-propyloxycarbonyloxymethyl ester); Asp-(cyclohexylcarbonyloxymethyi 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); and Asp-(1-(2-(2-methoxypropyl)carbonyloxy)ethyl ester). The present invention includes conpounds of formula (IV) : M x. \ / NR2 \ \ • (CHR')n °' (CH2)n.^^W R10 (IV) or a pharmaceutically acceptable salt form thereof/ wherein; R1 and R2 are independently selected from H or methyl; R10 is H, halogen/ Ci-Cs aUcyl, phenyl or C1-C4 alkoxy; n is 1; 263456 n' is 1; J is selected from: D-Val, D-2-aminobutyric acid, D-Leur D-Ala, D-Pro, D-Ser, D-Lys, 0-Ala, NMeGly, D-Nle, D-phenylGly, D-Ile, D-Phe, or D-Tyr; K is CCNMeArg; l is Gly; M is selected from the group consisting of: Asp-(methylcarbonyloxymethy ester); Asp-(ethylcarbonyloxymethyl ester); Asp-(t-butylcarbonyloxymethyl ester); Asp-(cyclohexylcarbonyloxymethyi ester); Asp-(1-(methylcarbonyloxy)ethyl ester); Asp-(1-(ethylcarbonyloxy)ethyl ester); Asp-(1-(t-butylcarbonyloxy)ethyl ester); A3p-(1-(cyclohexylcarbonyloxy)ethyl ester); Asp-(x-propyloxycarbonyloxymethyl ester); Asp-(cyclohexylcarbonyloxymethyi 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)-lr 3-dioxa-cyclopenten-2-one-4-yl)methyl ester) ; Asp-((1,3-dioxa-5-phenyl-cyclopenten-2-one-4-yl)methyl ester); and Asp- (1- (2 - (2-methoxypropyl) carbonyloxy) ethyl ester)". j PA^?Of?iCC[ ! -- -i i 263^56 The present invention includes compounds of formula (21a); (Ha) or a pharmaceutically acceptable salt form thereof, which is selected from the group consisting of: the compound of formula (Ila) wherein R^-, R2 and R10 are H; J is D-Val; K is NMeArg; L is Gly; and M is Asp-(methylcarbonyloxymethyl ester); the compound of formula (Ila) wherein R1, and R10 are H; J is D-Val; K is NMeArg; L is Gly; and M is Asp-(ethylcarbonyloxymethyl ester); the compound of formula (Ila) wherein R1, R^ and R10 are H; J is D-val; K is NMeArg; L is Gly; and M is Asp-(t-butylcarbonyloxymethyl ester); the compound of formula (Ila) wherein R^-, R^ and R1® are H; J is D-Val; K is NMeArg; L is Gly; and M is Asp- (cyclohexylcarbonyloxymethyi ester); the compound of formula (Ila) wherein R1, R2 and R10 are H; J is D-Val; K is NMeArg; L is Gly; and M is Asp-(1-(methylcarbonyloxy)ethyl ester); the compound of formula (Ila) wherein R*, r2 and R10 are H; J is D-Val; K is NMeArg; L is Gly; and M is Asp-(1-(ethylcarbonyloxy)ethyl ester); - 263*56 the compound of formula (Ila) wherein R1, R2 and R10 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 (Ila) wherein R^ and R10 are H; J is D-Val; K is NMeArg; L is Gly; and M is Asp-(1-(cyclohexylcarbonyloxy)ethyl ester); the compound of formula (Ila) wherein R^, R^ and R1® are H; J is D-Val; K is NMeArg; L is Gly; and M is Asp-(i-propyloxycarbonyloxymethyl ester); the compound of formula (Ila) wherein R1, R^ and R10 are H; J is D-Val; K is NMeArg; L is Gly; and M is Asp-(cyclohexylcarbonyloxymethyi ester); the compound of formula (Ila) wherein R1, R^ and R10 are H; J is D-Val; K is NMeArg; L is Gly; and M is Asp-(t-butyloxycarbonyloxymethyl ester); the compound of formula (Ila) wherein R*, R^ and R-0 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 (Ila) wherein R^, R^ and R10 are H; J is D-Val; K is NMeArg; L is Gly; and M is Asp-(1-(cyclohexyloxycarbonyloxy)ethyl ester); the compound of formula (Ila) wherein R^, and r10 are H; J is D-Val; K is NMeArg; L is Gly; and M is Asp-(l-(fc-butyloxycarbonyloxy)ethyl ester); the compound of formula <IIa) wherein R*r R^ and R10 are Hj J is D-Val; K is NMeArg; L is Gly; and M is Asp-(dimethylaminoethyl ester>; N.Z. PATENT OFFICE 1- OCT 1996 'received" 263456 the compound of formula (Ila) wherein R^, r2 and R10 are H; J is D-Val; K is NMeArg; L is Gly; and M is Asp-(diethylaminoethyl ester); the compound of formula (Ila) wherein R1, R2 and R10 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 (Ila) wherein R1, R^ and R10 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 (Ila) wherein R1, r2 and R10 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 (Ila) wherein R1, R2 and R10 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 (Ila) wherein R1, R2 and K10 are H; J is o-2-aminobutyric acid; K is NMeArg; L is Gly; and M is Asp-(methylcarbonyloxymethyl ester); the compound of formula (Ila) wherein Rl, R2 and R10 are H; J is c-2-aminobutyric acid; K is NMeArg; L is Gly; and M is Asp-(ethylcarbonyloxymethyl ester); the compound of formula (ila) wherein R*, R2 and R1^ are H; J is D-2-aminobutyric acid; K is NMeArg; L is Gly; and M is Asp-(t-butylcarbonyloxymethyl ester); the compound of formula (Ila) wherein r1, R2 and R10 are H; J is D-2-aminobutyric acid; K is NMeArg; L is Gly; and M is Asp-(cyclohexylcarbonyloxymethyi^ ester); 263456 the compound of formula (Ila) wherein R^, R2 and R10 are H; J is D-2-aminobutyric acid; K is NMeArg; L is Gly; and M is Asp-(1-(methylcarbonyloxy)ethyl ester); the compound of formula (Ila) wherein r1, r2 and r10 are H; J is D-2-aminobutyric acid; K is NMeArg; l> is Gly; and M is Asp-(1-(ethylcarbonyloxy)ethyl ester); the compound of formula (Ila) wherein R1, R2 and R^O are H; J is D-2-aminobutyric acid; K is NMeArg; h is Gly; and M is Asp-(l-(t-butylcarbonyloxy)ethyl ester); the compound of formula (Ila) wherein Ri, R2 and R10 are H; J is D-2-aminobutyric acid; K is NMeArg; L is Gly; and M is Asp-(1-(cyclohexylcarbonyloxy)ethyl ester); the compound of formula (Ila) wherein Ri, R2 and R1® 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 (Ila) wherein R1, R2 and R10 are H; J is o-2-aminobutyric acid; K is NMeArg; L is Gly; and M is Asp-(cyclohexylcarbonyloxymethyi ester); the compound of formula (Ila) wherein R^-, R2 and R10 are H; J is D-2-aminobutyric acid; K is NMeArg; L is Gly; and M is ASp-(t— butyloxycarbonyloxymethyl ester)s the compound of formula (Ila) wherein R^, R2 and R10 are H; J is D-2-aminobutyric acid; K is NMeArg; L is Gly; and M is Asp-(l-(i-propyloxycarbonyloxy)ethyl ester); the compound of formula (Ila) wherein R^, R2 and R10 are H; J is o-2-aminobutyric acid; K is NMeArg; L is Gly; and M is Asp- (1-(cyclohexyloxycarbonyloxy)ethyl ester); 263456 the compound of formula (Ila) wherein R1, R^ and R10 are H; J is D-2-aminobutyric acid; K is NMeArg; L is Gly; and M is Asp-(l-(fc-butyloxycarbonyloxy)ethyl ester); the compound of formula (Ila) wherein R1, R2 and R10 are H; J is D-2-aminobutyric acid; K is NMeArg; L is Gly; and M is Asp-(aimethylaminoethyl ester); the compound of formula (Ila) wherein R1, R2 an<^ rIO are h; J is D-2-aminobutyric acid; K is NMeArg; L is Gly; and M is Asp-(diethylaminoethyl ester); the compound of formula (Ila) wherein R1, R2 and R10 are H; J is D-2-aminobutyric acid; K is NMeArg; L is Gly; and M is Asp-((l/3-dioxa-5-methyl-cyclopenten-2-one-4-yl)methyl ester); the compound of formula (Ila) wherein R*, R2 and R10 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-or.e-4-yl) methyl ester) ; the compound of formula (Ila) wherein R1, R2 and R10 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 (Ila) wherein R^, R2 and R10 are H; J is D-2-aminobutyric acid; K is NMeArg; L is Gly; and M is Asp-(l-(2— (2-methoxypropyl)carbonyloxy)ethyl ester).

Hie present invention includes compounds of formula (I): N.Z. PATENT OFFICE 1 - OCT 1996 received 263456 or a pharmaceutically acceptable salt form thereof wherein: R1 is Rf C1-C4 alkyl, phenyl, benzyl or phenyl (C3.-C4) alkyl; R2 is H or methyl; R10 is H, halogen, Ci-Cs alkyl, phenyl or C1-C4 alkoxy; n is 0-2 ; n' is 0-1; J is Ala, Val, lie, Leu, Nle, phenylGly, Phe, Lys, Orn, Met, Pro, 0-Ala, Tyr, Ser, NMeGly, cyclohexylGly, cyclohexylmethylGly, ncrvaline, 2-aminobutyric acid, 2-aminopentanoic acid/ Gly, Cys, S-benzyl-Cys, S-raethyl-Cys, Asp, Glu, 2-amino-2~ methylpropionic acid, His, 1-allo-isoleucine, Asn, Gin, Thr, Trp, or O-methyl-Tyr; K is Arg, N5-Me-N8-guanidinylOrn, . p~aminomethylPhe, p-guanidinylPhe, Lys or N8-MeLys; L is Gly; r M is selected from the group consistingj^&ji PATENT ofncej 263 45 Asp-(methyicarbonyloxymethyl ester); Asp-(ethylcarbonyloxymethyl ester)? Asp-(t-butylcarbonyloxymethyl ester); Asp-(cyclohexylcarbonyloxymethyi 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-(cyclohexylcarbonyloxymethyi 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); and Asp-(1-(2-(2-methoxypropyl)carbonyloxy)ethyl ester). 263456 Ihe present invention includes ccrpounds of formni^ (xx).

Rio (It) or a pharmaceutically acceptable salt form thereof wherein: R1 is H, C1-C4 alkyl, phenyl, benzyl or phenyl (C1-C4)alkyl; R2 is H or methyl; R10 is H, halogen, Ci-Cg alkyl, phenyl or C3.-C4 alkoxy; alternatively, when R10 is para to the carbonyl, R10 and R1 may be taken together to form -CH2-CH2-CH2- thereby to form a six-membered fused ring; n is 0-1; J is Ala, Val, lie, Leu, Nle, phenylGly, Phe, Lys, Orn, Met, Pro, (J-Ala, Tyr, Ser, NMeGly, cyclohexylGly, cyclohexylmethylGly, norvaiine, 2-aminobutyric acid, 2-aminopentanoic acid, Gly, NE-p~azidobenzoyl-Lys, Cys, S-benzyl-CyS/ S-methyl-Cys, Asp, Glu, 2-amino-2~methylpropionic acid, His, 1-allo-isoleucine, Asn, Gin, Thr, Trp, or O-methyl-Tyr; K is Arg, N8-Me-N8-guanidinylOrnf Lys or Ne-MeLys; L is Gly; and M is selected from the group consisting of: 26 3 4 5.6 Asp-(methyicarbonyloxymethyl ester); Asp-(ethylcarbonyloxymethyl ester); Asp-(t-butylcarbonyloxymethyl ester); Asp-(cyclohexylcarbonyloxymethyi 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-(cyclohexylcarbonyloxymethyi 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-(diethylaminoetliyl 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); and Asp-(1-(2-(2-methoxypropyl)carbonyloxy)ethyl ester).

Ihe present invention includes compounds of formula (II), or a pharmaceutically acceptable salt thereof, wherein: n is 1; K is Arg; j is Ala, Val, lie, Leu, Pro, Ser, or Lys.

N.Z PATENT OfnCE l- m m WO 94!22910 26T^o In the present invention it has been discovered that the compounds above are useful as inhibitors of glycoprotein Ilb/IIIa (GPIIb/IIIa). As discussed above, 5 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. 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 15 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 20 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 25 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 30 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 35 boropeptides, hirudin or argatroban. The compounds of the present invention may also be combined or The ccnpounfls of the present invention are useful for the H.Z. p 26 3456 2C 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 (PAI- 1) or the dense granule (serotonin) are not affected by the GPIIb/IIIc. 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-1). 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 < 0.01 (AM) 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 disorder's. Such wo 94/22910 ZBTf? 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 10 (LMWH), including pharmaceutically acceptable salts or prodrugs thereof. For reasons of efficacy, the preferable anti-coagulant agents are warfarin or heparin or LMWK. The warfarin employed herein, may be, for example, crystalline warfarin or amorphous sodium 15 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 20 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, 25 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 30 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 35 pharmaceutically acceptable salts or prodrugs thereof. Ticlopidine is also a preferred compound since ..i£ is 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 5 pharmaceutically acceptable salts or prodrugs thereof.

/ WO 94/22910 - 29- PCT/US94/03223 26345 6 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 5 thrombin-mediated platelet activation (that is, for example, the aggregation of platelets, and/or the granular secretion of plasminogen activator inhibitor-1 and/or serotonin) and/or fibrin formation are disrupted.

Such inhibitors include boropeptides, hirudin and 10 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. ,187,15*7 and European Patent Application Publication Number 293 881 A2, the disclosures of which are hereby incorporated herein by reference. Other suitable 25 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. r H.Z * i ; v "" '■ 1 26 3 A 5 6 Tissue plasminogen activator (tPA) is commercially available from Genentech Inc., South San Francisco, California. The term anistreplase, as used herein, refers to anisoylated plasminogen streptokinase 5 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, 10 Middlesex, England, under the trademark EMINASE™. 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. 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 20 permitting the use of lower doses of each. A lower dosage 1r1ir.ir.i2es 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 25 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 anticoagulants significantly reduced overall mortality and 30 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 35 additional study, the Warfarin Reinfarction Study, has also recently demonstrated a significant,reduction in Combination products, where the cyclic N.Z. PATENT OfflCE I 1 - nr,T 1996 PCT/U§94/03223 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 5 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 10 large, randomized, double-blind, placebo-controlled clinical studies. In addition, ASA has been approved for us« in the secondary prevention of myocardial infarction, bas^d on data from several trials which, when pooled, suggested a reduction in reinfarction and 15 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 20 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 25 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 30 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 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 a:i anti-coagulant agent such as warfarin or heparin, or an antiplatelet agent such as aspirin, pirojcicsjm 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 ir 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 PCTAJS94/03223 invention, or combinations thereof, are directed to meeting these, as well as other, needs.

GPIIb/IIIa is known to be overexpressed in 5 metastatic tumor cells. The compounds or combination products of the present invention may also be useful for the treatment, including prevention, of metastatic cancer. 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 15 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 20 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 25 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, 30 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 3-35 letter abbreviation of the amino acid, as indicated by the following examples: D-Leu, D-Leu, L-Leu,„ or L-Leu\ The compounds herein described may have asymmetric When any variable (for example, R1 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 R*1, then said group may optionally be substituted with up to two R11 and R11 at each occurrence is selected independently from the defined list of possible R11. 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 reactior. 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 WO 94/22910 - 35- PCT/US94/03223 263456 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+l) ) ; "alkoxy" represents an alkyj group of indicated number of carbon 5 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" 10 is intended to include saturated bicyclic ring groups such as (3.3.OJbicyclooctane, [4.3,0]bicyclononane, [4.4.OJbicyclodecane (decalin), [2.2.2]bicyclooctane, and so forth. "Alkenyl" is intended to include hydrocarbon chains of either a straight or branched 15 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 20 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 25 group of the formula ~B(r34)(r35)t wherein R^4 and R3^ are independently selected from: -OH; -F; -NRi3r14. or Ci-C8~alkoxy; or R3^ and R^S 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 chair, or ring contains from 2 to 20 carbon atoms and, optionally, 1-4 heteroatoms independently selected from N, S, or 0; a cyclic boron amide-ester where said chain 35 or ring contains from 2 to 20 carbon atoms and, optionally, 1-4 heteroatoms independently seles^ed from WO 94/22910 _36_ PCT/US94/03223 263456 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 R11.

Boron esters include boronic acid protecting 5 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, 10 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 20 like.

As use.-S herein, "aryl" or "aromatic residue" is intenaad 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 25 or 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, 30 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 10-35 membered bicyclic heterocyclic ring which may be saturated, partially unsaturated, or aromatic, and which 263 456 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 5 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 10 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, 15 indolene, quinoline, isoquinolinyl or benzimidazolyl, piperidinyl, 4-piperidone, 2-pyrrolidone, tetrahydrofuran, tetrahydroquinoline, tetrahydroisoquinoline, decahydroquinoline, octahydroisoquinoline, azocine, triasine (including 20 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, 2/f-pyrrole, pyrrole, imidazole, pyrazole, thiazole, 25 isothiazole, oxazolc' ;including 1,2,4- and 1,3,4-oxazole), isoxazole, triazole, pyridine, pyrazine, pyrimidine, pyridazine, indolizine, isoindole, 3H-indole, indole, l#-indazole, purine, 4H-quinolizine, isoquinoline, quinoline, phthalazine, naphthyridine, 30 quinoxaline, quinazoline, cinnoline, pteridine, 4atf-carbazole, carbazole, A-carboline, phenanthridine, acridine, perimidine, phenanthroline, phenazine, phenarsazine, phenothiazine, furazan, phenoxazine, isochroman, chroman, pyrrolidine, pyrroline, 35 imidazolidine, imidazoline, pyrazolidine, pyrazoline, piperazine, indoline, isoindoline, quinuclidine, or WO 94/22910 - 38- PCT/US94/03223 263^5 6 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 hydroxy1, 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 10 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, 15 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 R^1, C1-C6 alkylaminocarbonyl substituted with 0-3 benzoyl substituted with 0-3 R^, phenoxycarbonyl substituted with 0-3 phenylaminocarbonyl substituted with 0-.? r!2. Examples of groups that, when administered to a mammalian subject, are cleaved to form a free hydroxyl, amino or sulfhydryl, include hydroxy, amine or 25 sulfhydryl protecting groups, respectively.

As used herein, the term "amine protecting group" means any group known in the art of organic synthesis 30 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 35 (1981), the disclosure of which is hereby incorporated by reference. Any amine protecting group known in the WO 94/22910 ~39~ ^ PCT/US94/03 263 art can be used. Examples of amine protecting groups include, but. are not limited to, the following: 1) acyl types such as formyl, trifluoroacetyl, phthalyl, and p-toluenesulfonyl; 2) aromatic carbamate types such as 5 benzyloxycarbonyl (Cbz) and substituted benzyloxycarbonyls, 1-(p-biphenyl)-1- methylethoxycarbonyl, and 9-fluorenylmethyloxycarbonyl (Fmoc); 3) aliphatic carbamate types such as tert-butyloxycarbonyl (Boc), ethoxycarbonyl, 10 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 15 containing types such as phenylthiocarbonyl and dithiasuccinoyl. Also included in the term "amine protecting group" are acyl groups such as azidobenzoyl, p-benzoylbenzoyl, o-benzylbenzoyl, p-acetylbenzoyl, dansyl, glycyl-p-benzoylbenzoyl, phenylbenzoyl, 20 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). 25 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 35 functional groups present in the compounds in such a way that the modifications are cleaved, either in routine ^54 5 6 manipulation or in viso, to the parent cj 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 15 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 Remington's Pharmaceutical Sciences, 17th ed.. Mack 20 Publishing Company, Easton, PA, 1985, p. 1418, the disclosure cr 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 25 modified and unusual amino acids, such as those disclosed in, for example, Roberts and Vellaccio (1983) Thp 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 30 include, but are not limited to, D-amino acids, hydroxylysine, 4-hydroxyproline, ornithine, 2,4-diaminobutyric acid, hoir.oarginine, norleucine, N-methylaminobutyric acid, naphthylalanine, phenylglycine, fl-phenylproline, tert-leucine, 35 4-aminocyclohexylalanine, N-methyl-norleucine, 3,4-dehydroproline, 4-aminopiperidine-4-carboxylic acid, WO 94/22910 PCT/US94/03223 263 456 6-aminocaproic acid, trans-4-(aminomethyl)-cyclohexanecarboxylic acid, 2-, 3-, and 4-(aminomethyl)-benzoic acid, 1-aminocyclopentanecarboxylic acid, 1-aminocyclopropanecarboxylic acid, and 2-benzyl-5-5 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 10 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 15 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 2 0 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 30 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 35 substitute or amide "equivalent" linkages are formed from combinations of atoms not normally found in WO 94/22910 - 4 2- PCT/US94/03223 263456 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: o-Abu (J-Ala, b-Ala or 15 pala Eoc Boc-iodo-Mamb Boc-Mamb B;r-ON Cl2Bzl CEZ or Cbz 25 DCC DIEA di-NMeOrn DMAP HBTU NMeArg or MeArg NMeAmf NMeAsp NMeGly or MeGly D-2-aminobutyric acid 3-aminopropionic acid t-butyloxycarbonyl t-butyloxycarbonyl-3-aminomethyl~4-iodo-benzoic acid t-butyloxycarbonyl-3-aminomethylbenzoic acid f2-(tert-butyloxycarbonyloxyliminco -2- phenylacetonitrile dichlorobenzyl Ca rboben z y1oxy dicyclohexylcarbodiimide diisopropylethylamine N-aMe-N-7Me-orni thine 4-dimethylaminopyridine 2-(lH-Benzotriazol-l-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate a-N-methyl arginine N-Methylaminomethylphenylalanine a-N-methyl aspartic acid N-methyl glycine N.Z. PATENT OfFVCE 1- OCT RECLiVED 263 45 6 NMe-Mamb N-methyl-3-aminomethylbenzoic acid NMM N-methylmorpholine OcHex O-cyclohexyl OBzl O-benzyl TBTU 2-<lH-Benzotriazol-l-yl)-1,1,3,3- tetramethyluronium tetrafluoroborate Tos tosyl The following conventional three-letter amino acid 10 abbreviations are used herein; the conventional one-letter amino acid abbreviations are not used herein: Ala - alanine Arg - arginine Asn m asparagine Asp - aspartic acid Cys - cysteine Gin - glutamine Glu m glutamic acid Gly - glycine His - histidine lie - isoleucine Leu - leucine Lys u lysine Met o methionine Nle - norleucine Phe - phenylalanine Phg JC phenylglycine Pro - proline Ser « serine Thr - threonine Trp m tryptophan Tyr - tyrosine Val - valine Peptide Synthesis [FTz V* "" \ rftswfo t • » +-■*. '\',N ? 26 3 A 5 6 The compounds of the present invention can be synthesized using standard synthetic methods known to those skilled in the art. Preferred methods include but 5 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 10 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 15 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, 25 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); and Bodanszky et al. "The Practice of Peptide Sythesis" 30 Springer-Verlag, New York (J.984) , 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 35 standard coupling procedures such as the azide method, mixed carbonic acid anhydride (isobutyl chloroformate) N.Z. PATENT OfFjCtE 1- OCT 1996 WO 94/22910 - 45~ PCT/US94/03223 263456 method, carbodiimide (dicyclohexylcarbodiimide, diisopropylcarbodiimide, or water-soluble carbodiimides) method, active ester (p-nitrophenyl ester, N-hydroxysuccinic imido ester) method, Woodward reagent K 5 method, carbonyldiimidazole method, phosphorus reagents such as BOP—CI, or oxidation-reduction method. Some of these methods (especially the carbodiimide) can be enhanced by the addition of 1-hydroxybenzotriazole.

These coupling reactions may be performed in either 10 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, 15 "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) 25 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 30 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 35 aminomethyl resin. Many of these resins are 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), «»thoxycarbonyl, diisopropylmethoxycarbonyl, and allyloxycarbonyl; 4) cyclic alkyl carbamate types such as cyclopentyloxycarbonyi 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 B^>c 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 HC1 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.

\V0 94/22910 PCT /US94/03223 263456 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 5 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 10 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 15 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-20 methoxybenzyl, acetamidomethyl, benzyl, or t- butylsulfonyl for cysteine; and the indole of tryptophan can either be left unprotected or protected with a formyl group. 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. 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 When Fmoc is chosen for the a-amine protection Once the elongation and cyclization of the peptide y. ■ i WO 94/22910 PCT/US94/03223 263 456 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-5 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 10 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 15 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 20 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 25 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) 30 J. Org. Chem. 48, 77 (1982)), which are incorporated here by reference.

The compounds o£ the present invention may be prepared using the procedures further detailed below as 35 well as the procedures described in PCT Patent Application International Publication Number 263 45 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 5 Publication Number WO 93/07170. used in preparing the compounds of the invention are described further below.

Manual solid phase peptide synthesis was performed 10 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 15 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 20 Laboratories (Belmont, CA), or Sigma (St. Louis, MO). 2-(lH-Benzotriazol-l-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, 25 diisopropylethylamine (DIEA), 3-cyanobenzoic acid and [2-(tert-butyloxycarbonyloxylimino)-phenylacetonitrile] (Boc-ON) were purchased from Aldrich Chemical Company. Dimethylformamide (DMF), ethyl acetate, chloroform (CHCI3)t methanol (MeOH), pyridine and hydrochloric acid 30 (HC1) 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 35 purchased from Fluka Chemical Company. Absolute ethanol was obtained from Quantum Chemical Corporation. Thin Representative materials and methods that may be N.Z. PATENT OFFICE 1- OCT 1996 layer chromatography (TLC) was perfofflecr^i^illccTGe 60 F254 TLC plates (layer thickness 0.2 mm) which were purchased from EM Separations. TLC visualization was accomplished using UV light, iodine, and/or ninhydrin 5 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 10 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 nnri d-substH tuted Boc-aminomgfhylbenzoic Acid Derivatives 3 and 4-substituted Boc-aminomethylbenzoic acid 20 derivatives useful as intermediates in the synthesis of the compounds of the invention are prepared using standard procedures, for example, as described in Tett. Lett., 4393 (19*75); Modern Synthetic Reactions, H.O. House (1972); or Harting et al. J. Am. Chem. Soc., 50: 25 3370 (1928) , ar.d as shown schematically below.

CN r -nf NHz~~HCI BO&OW BOC 1 Et)N,pHct " ' 3-Amlnomethvlbenzoic acid»Hrl 3-Cyanobenzoic acid (10.0 g, 68 mmol) was dissolved in 200 ml ethanol by heating in a 35-50°C water bath.

N.Z. PATENT OFFICE 1- OCT 1996 ^ RECEIVED J WO 94/22910 PCT/US94/03223 263456 Concentrated HC1 (6.12 ml, 73 mmol) was added and the solution was transferred to a 500 ml nitrogen-flushed round bottom flask containing palladium on carbon catalyst (1.05 g, 10% Pd/C). The suspension was stirred 5 under an atmosphere of hydrogen for 38 hours, filtered through a scintered glass funnel, and washed thoroughly with H2O. The ethanoi was removed under reduced pressure and the remaining aqueous layer, which contained a white solid, was diluted to 250 ml with 10 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 15 lyophilized to give the title compound (3- aminomethylbenzoic acid-HCl) (8.10 g, 64%) as a beige solid. 3-H NMR (D20) 4.27 (s, 2H) , 7.60 (t, 1H) , 7.72 (d,1H), 8.06 (d, 2H). t—Butvloxvcarbony1—3-aminomethvlbenzoic Acid (Boc-Mamb) The title compound was prepared according to a modification of standard procedures previously reported in the literature (Itoh, Hagiwara, and Kamiya (1975) 25 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 irnnol) in 60 ml acetone followed by triethylamine (5.56 ml, 39.9 mmol). The solution 30 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 35 was washed three times with ether. The aqueous layer was then acidified to pH 2 with 2N HC1 and then WO 94/22910 — «— A PCT/US94/03223 263^56 extracted three times with ethyl acetate. The combined organic layers were washed three times with H2O, 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—125°C ;iH NMR (CDCI3) 1.47 (s, 9 H) , 4.38 (br s, 2 H), 4.95 (br s, 1H), 7-45 <t, 1H) , 7.55 (d, 1H), 8.02 (d, 2K). t-Butvloxvcarbonyl-N-methvl-3-aminoniethvl hpn7.olr Acid (Bon-NMftMamb) The title compound can be prepared according to 15 standard procedures, for examples, as disclosed in Olsen, J. Org. Chem. (1970) 35: 1912), and as shown schematically below.

BOC-NH' B0°-N °He CHjl DMF, 45° Synr.hftfiifi of ftmlnomfit.hylbenzoif; ftctd Analogs Intermediates of the formula below may be prepared 25 using standard synthetic procedures, for example, as shown in the indicated reaction schemes shown below. -n . : . • v:;-.

W. 263 NH-BOC For R - CH3, CH2CH3, CH2CH2CH3, CH2CH2CH2CH3, CH (CH3) 2, C (CH3) 3, CH (CH3) CH2CH3, benzyl, cyclopehtyl, cyclohexyl; see Scheme 1.

For R » CH3, CH2CH2CH2CH3, phenyl; see Scheme 2. For R - CH3, phenyl; see Scheme 3 and 4.

IN.Z. PAjeffcM wo 94/22910 PCT/us94/03223 ■263456 Scheme, 1: 9 ^ O 10% Pd-C/Hy o NH,.HCI """"n"™ . ".o-Vy ' Ptl HN«C Ph C^Cij HO Ph Ph J pj-^ O NH-BOC (') D»pw»Oion Q K^Ph 0 H -2L!^ m«JW.r Schemf? 2; kl O a A tSiMtj N^0> HzS°4 at»/A JL J NaN(SiMe3)2 ■ n kJ - Ha\j lOr 1(1) RLi |(ii) HgO/HCl O NH-BOC SOOON § NHg.HC' HO ~ „ , autK^N II I ' >J^R "0^0^.

Frheme 3: o o hoAOa' NH4OAc/Na(CN)BH3 CH3OH/M0I. Sieves 263 45 6 O NH2HCI h°aTQTAr Boc-On/Acetone NH-BOC R Scheme 4: HO 0) NH2OH.HCI/EtOH/Pyr. j? NH^CI ' YT r ho^YT R (ii) Pd-C/EtOH/HCIcon./H2 l|^J Boc-On/Acetone NH-BOC 3- fl (r-butvloxvparbonvl) amino!ethvlbenzoic arid (BQC-MeMftMB) 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".p| 94.4% yield).

N.Z. PATENT OfflCg 1- or.T 199$ ihnmr (CD3OD) 7.4 5-8.30(111, 4h) , zt30 (st 3h) . ms ICH4-ci) [m+h-o] - 164.

A solution of the oxime (0.51 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 Hz 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). ^HNMR (CD3OD) 10 7.6-8.15(m, 4H), 4.55(q, 1H) , 1.70 (s, 3H) . MS [M+H] » 166.

Amine hydrochloride {0.40 g, 2 nunol) 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 15 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 20 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 (CDCI3) 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-M (t-h^yTrtvyciarhonvl ^amino!benzvlhengpjr arid (BOC-PhMAMB) The title compound for the purpose of this 30 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 35 pyridine (2.00 ml, 25 mmol) in ethanol was refluxed for 12 h. After the usual extractive work up, white solid [ntpatentot^ 1- OCT 1996 1 RECEIVED ~ J 263 45 < ic^s^-Ar contain* was obtained (2.41 g) . The produc traces of pyridine, but was used in the next step without further purification.

The crude product (2.00 g, -S mmol) was dissolved 5 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 10 vacuum, amine hydrochloride was obtained as a white solid (2.12 g ; 97% yield). ^-HNMR (CD3OD) 7.4-8.15 (m, 10H}, 5.75 (s, 1H) . 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 15 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. ^HNMR (CD3OD) 7.2-8.0(m, 10H), 5.90 (2s, 1H, 2 isomers), 1.40(s, 9H). MS (NH3-CI) [M+NH4-C4H8] = 289 t-Butvloxycarbonvl-D-2-aminobutvric Acid The title compound was prepared by a modification of procedures previously reported in the literature 25 (Itoh, Hagiwara, and Kamiya (1975) Tett. Lett., 4393), as shown in the scheme below.

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 IN.Z PA7OT' i— i \ 1$$ \ -58- PCT/US94/03223 '263456 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 5 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-10 butyloxycarbonyl-D-2-aminobutyric acid as an oil (2.05 g,greater than quantitative yield, contains solvent), which was used without further purification. ^H NMR (CDCI3) 0.98 (t, 3H), 1.45 (s, 9H), 1.73 (m, 1H), 1.90 (m, 1H), 4.29 (m, 1H), 5.05 (m, 1H).

Synthesis of t-BntvTnxvcarbonvl-3-aminophanvlacerir Acid t-Butyloxycarbonyl-3-aminophenylacetic acids useful as intermediates in the synthesis of the compounds of 20 the invention are prepared using standard procedures, for example, a: described in Collman and Groh (1982) .7 Arc. Chem. Soc., 104: 1391, and as shown schematically below. f«^rsY^XC02H fS^f^COaH Cr~ CP J F«SO««7HaO J l-BuOjCOCOjt-Bu OlEA • N02 H H H' COjt-Bu t-Butvloxvcarbonvl-3-aminophenvlacetic 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 f OCT !t~—"iicav®. J PCTAJSS . temperature. The reaction mixtux partitioned between dichloromethane-H20, the water layer was separated, acidified to pH 3 with IN HCl, and extracted with dichloromethane. The extracts were washed 5 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 105°C; NMR (CDCI3) 7.35 (s, 1H), 7.25 (m, 10 3H), 6.95 (m, 1H), 6.60 (br s, 1H), 3.65 (s, 2H), 1.50 (s, 9H) .

Synthesis of 4. S. and 6-Subsfcltuf.gd 3-AminQmsthvlhengni r Acid«HCl. and 4. 5. and 6-Subst.ituted 15 t-Butvloxvcarhnnvl-Vaminomethvlbenzolc Acid Derivatives 4, 5, and 6-Substituted 3-aminomethylbenzoic acid*HCl, and 4, 5, and 6-substituted t-butyloxycarbonyl-3-aminomethylbenzoic acid derivatives 20 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.

Chiin. 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.

£ HZ. PATENT Of- ; CO2H X BOC-HN "Xp. 263456 CI2CHCONHCHjOH conc H2S04 CI y\ CI C02H Boc-ON, EtjN Synthesis of 4-Chloro-3-aminomethvlhenzoir Acid«HCl The title compound was prepared by modification of procedures previously reported in the literature (Felder et al (1965) Helv. Chim. Acta, 48: 259). To a solution of 4-chlorobenzoic acid (15.7 g, 100 mniol) in 150 ml of 10 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 ica, stirred for 1 hour, the solid was collected by filtration, and washed with H2O. 15 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 H2O, and air-dryed overnight to give 4-chloro-3-dichloroacetylaminomethylbenzoic acid (26-2 g, 20 89%) as a white powder.

A suspension of 4-chloro-3-dichloroacetylaminomethylbenzoic acid (26.2 g, 88 mmol) in 4 5 ml of acetic acid, 150 ml of concentrated HCl, and [N"Z~PATENTOFBCt] OCT ^".iVED 263456 150 ml of H20 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 5 give the title compound (7.6 g, 39%) as off-white crystals, mp 278-9'C; Ifi NMR (D6-DMSO) 13.40 (br s, 1H) , 8.75 (br s, 3H), 8.20 (s, 1H), 7.95 (dd, 1H), 7.70 (d, 1H), 4.20 (br s, 2H) . t-Butvloxyearbonyl —4-fh 1 orn-.V«minomi»thy1 hi»riy.filp Acid 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 H20, was added to a solution of Boc-ON 15 (9.2 g, 38 mmol) in 50 ml of tetrahydrofuran cooled to 0°C. 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, 20 acidified to pH 3 with IN KC1, 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 25 compound (7.4 g, 87%) as a white powder, mp 159°C (dec); NMR (Dg-DMSO) 13.20 (br s, 1H), 7.90 (s, 1H), 7.80 (dd, 1H), 7.60 (br s, 1H), 7.55 (d, 1H), 4.20 (br d, 2H) , 1.40 (s, 9H) . 4 anH 6-Suhsf i f uteri t.-Bnt vloxvrarhonyl-3- The compounds listed below were prepared using the general procedure described above for t-35 butyloxycarbonyl-4-chloro-3-aminomethylbenzoic acid. aminomethylhenzoic Acid Derivatives —'yv*' 263456 plO p10a B10a £10 mp °C H CI 159 H I 168 H Me 155 H MeO 171 CI H 150 I H 182 Me H 166 MeO H 79 Synthesis of ?-Am^nmf»thy1benznir ft.ciri»KC1 and 2-5 Aro-i nnm»t fry 3 phgny 1 ace11 r. AcId « HCl 2-Aminomethylbenzoic acid-HCl and 2-aminomethylphenylacetic acid*HCl useful as intermediates in the synthesis of the compounds of the invention are C prepared using standard procedures, for example, as described in Naito et al J. Antibiotics, 30: 698 (1977); or Young and Sweet J. Am- Chem. Soc., 80: 800 (1958), and as shown schematically below.

PCT/U594/03223 2-Ami nnmPfhylphgnvl aret. ic Ac 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 5 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 30-40°C. After an additional 3 hours, the reaction mixture was 10 poured onto ice, and the water layer was made basic with concentrated ammonium hydroxide. The chloroform layer was separated, washed with H20, brine, dried over anhydrous magnesium sulfate, and evaporated to dryness under reduced pressure. This material was purified by 15 sublimination (145°C, <1 mm), followed by recrystallization from benzene to give the title compound (5.4 g, 45%) as pale yellow crystals, mp 14 9-150°C; XH NMR (CDCI3) 7.20 (m, 5H) , 4.50 (s, 2H) , 3.60 (s, 2H). 2-Amlnornf>t-hv:iphgnvlarftt:ir 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 2-25 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 30 provided the title compound (5.5 g, 62%) as colorless crystals, mp 168°C (dec); NMR (D6-DMSO) 12.65 (br s, IK), 8.2S (br s, 3H), 7.50 (m, 1H), 7.35 (m, 3H), 4.05 (ABq, 23), 3.80 (s, 2H). 2-Amlnomefhvlbenzoic Asld g-Larl-atn 1- -64-26345£r~23 The title compound was prepared by modification of procedures previously reported in the literature (Danishefsky et al . (1975) J". Org. Chem., 40: 796). A mixture of methyl o-toluate (45 g, 33 mol), N-5 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 150°C; NMR (CDCI3) 7.90 (d, 1H), 7.60 (t, 1H), 7.50 (t, 2H), 7.00 (brs, 1H), 4.50 (s, 2H) . procedure described above for 2-aminomethy Iphenylacet i r. acid*HCl. The lactam (3.5 g, 26 mmol) was converted to the title compound (2.4 g, 50%) as colorless crystals, mp 233°C (dec); 3-H NMR (Dg-DMSO) 13.40 (br s, 1H) , 8.35 25 (br s, 3H), 8.05 (d, 1H), 7.60 (m, 3H), 4.35 (br s, 2H).

Alternatives to Mamh: 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 35 the synthesis of cyclic peptides of the present invention are 2 0 2—Ami nomet.hvIbenzoic Acid»HCl ; compound was prepared using the general fnsa 2634 3 0 rctlvSM"aza described below. The synthesis of these intermediates is outlined below in Scheme 4a.

Scheme 4a ElOH "tSOjEt O n CHjC HSO, A 87% OOjEt Aids AoatyiO EDC 88% NaOH. BOH HjO* 00% CHtC NaOCt. NaOH HCL 95% jxrr- MO* Nad A 85% N*»c NHjOH-HCI O 72% 1,06 09 ' N-OH 1-yPd-C M«OHHO 27% °r ■n 7\ B » HHj «J>4I • HQ 1 ,)TMSCN / Znl; in B«nnn« 2.) POQ.n / Pyridine; rrftu* 47% CHjNH-MIOC 1.)HypJ-c MaOHWCI 2.) Saponify S 3.)AW-BOC PRMd 12 1.)Hj/Pd-C IMHtCI Z)Mf-BOC 3.) Saponify H CHtNH*««OC 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) .

I f- Wv>n ■ -66- PCT/US94/03223 26345 R—Ami no—5 . fi. 7 . fl-tptrahvdro-2-napht.hol e Acid Hydrochloride 18) As shown below in Scheme 4a, 4-phenylbutyric acid 5 (l) 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 10 group was oxidized to give 4-carboxyphenylbutyric acid (5) which was converted to the l-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 15 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 20 the cyclic peptide.

Part A - A solution of 4-phenylbutyric acid (50.0 g, 0.3 mol) in ethanol (14 0 mL) with concentrated sulfuric acid (0.53 mL) was stirred at reflux over 5 hours. The cooled 25 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 30 mol, 97%) as a yellow liquid. 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).

Part B - To a solution of aluminum chloride (153 g, 1.15 mol), and acetyl chloride (38.5 mL, 42.5 g, 0.54 mol) in OCT 1998 "received WO 94/22910 PCT/US94/03223 263456 dichloromethane (1500 mL) was added, dropwise, a solution of 4-phenylbutyric acid ethyl ester (50.0 g, 0.26 mol) in dichloromethane (500 mL). All was stirred at ambient temperature for 15 minutes. The solution was 5 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 4-10 acetylphenylbutyric acid ethyl ester (53.23 g, 0.23 mol, 88%) as a dark yellow liquid. 1H NMR (CDCI3) 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, 15 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 (IN). 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 - 50-52°C; *H NMR (CDCI3) 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).

Part 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 55°C, was added, portionwise as a solid, 4-acetylphenylbutyric acid (16.0 g, 0.078 mol) while keeping the temperature between 60-70°C. All was 35 stirred at 55°C over 20 hours. The cooled solution was quenched by the dropwise addition of a solution of 263456 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 4-carboxyphcnylbutyric acid (15.31 g, 0.074 mol, 95%) as a white solid, mp ■» 190-195°C; 1H NMR (DMSO) d 12.55 (bs, 1H), 8.1 (s, 1H), 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 E - 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 190°C over 30 minutes. As the mixture cooled to 60°C, cold hydrochloric acid (IN, 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 « 210-215°C; XH NMR (DMSO) d 8.4 (s, 1H), 8.1 (d, 2H, J=B.O Hz), 7.5 (d, 1H, 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).

Part 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, -- 263TP 0.0038 mol, 72%) as a white solid, mp « 205-215°C; 1H NMR (DMSO) d 11.3 <s, 2H), 8.4 (s, 1H) , 7.8 (d, 1H, J=7.7 Hz), 7.3 (d, 1H, J~7.7 Hz), 2.8 (t, 2H, J=5 . 9 Hz), 2.7 (d, 2H, 0=6.6 Hz), 1.9-1.7 (m, 2H).

Part G - A mixture of l-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) 10 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 15 chromatography using hexanerethyl 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 = 289-291°C; NMR (DMSO) d 8.55 (bs, 3H), 8.2-8.1 (m, 1H), 7.85-7.8 (m, 1H), 7.35-7.25 20 (m, 1H), 4.5 (m, 1H), 2.9-2.8 (m, 2H), 2.1-1.9 (m, 3H), 1.85-1.7 (m, 1H).

PCT /US94/03223 26345 N- (BOC) -8—Aminomp.t.hyl-5. 6.7. 8-tetrahvdro-2-napht:hoic ftrciri (12) As shown below in Scheme 4a, the remaining tetralone was then converted to the methyl ester (9) . 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 10 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 15 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.

Part A - Admixture of l-tetralon-7-carboxylic acid (7.C c, C.037 rfcl) 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 2 5 ana 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 30 acetate::75:25 . The resulting solid was triturated w-th hexane to give l-tetralon-7-carboxylic acid methyl ester (3.61 g, 0.018 mol, 49%) as a yellow solid, mp - 170-172°C; *H NMR (CDC13) d 8,7 (s, 1H) , 8.15 (d, 1H, J-8.1 Hz), 7.35 (d, 1H, J«8.1 Hz), 3.95 (s, 3H) , 3.05 (d, 2H, 35 J=6.1 Hz), 2.7 (t, 2H, J=6.4 Hz), 2.15 (quintet, 2H, J=e.2 Hz). -71 263456 •10 Part B - A solution of l-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 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 = 73-75°C; XH NMR (CDCI3) d 8.0-7.9 (m, 1H), 7.3-7.2 (m, 1H) , 6.95 (t, 1H, J-4.8 Hz), 3.95 (s, 3H) , 2.9 (t, 2H, J-8.3 Hz), 2.6-2.4 (m, 3H) Part 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 * 172-179°C; XH NMR (DMSO) d 8.2-8.0 <m, 4H), 7.9-7.7 Jit., 6H) , 7.5-7.2 (m, 4H) , 3.9-3.8 (m, 7H) , 3.3-2.7(m, 10H), 2.0-1.6 (m, 8H).

Part 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 t I 26 3 wrr tetrahydrofuran (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 5 concentrated to half volume and extracted with diethylether. The aqueous layer was then acidified to a pH of 1.0 using hydrochloric acid (IN) and then extraced with ethyl acetate. The combined organic layers were dried over anhydrous magnesium sulfate and evaporated to 10 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 - 72-80°C; XH NMR (DMSO) d 13.8 (s, 1H), 7.8-7.65 (m, 15 3H) , 7.6-7.5 (m, 3H), 7.25-7.20 (m, 1H), 7.15-7.05 (m, 1H), 3.9-3.8 (m, 1H), 3.2-2.8, (m, 4H), 1.8-1.6 (m, 3H), 1.4 (s, 6H) .

Part E - To a solution of methyl N-(BOC)-8-aminomethyl-20 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 25 equal to 1.0 using hydrochloric acid (IN). 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, 8-30 tetrahydro-2-naphthoic acid (0.19 g, 0.00062 mol, 39%) as a white solid, mp - 172-176°C; *H NMR (DMSO) d 7.8 (s, 1H), 7.65 (d, 1H, J-8.1 Hz), 7.15 (d, 1H, J»8.1 Hz), 7.1-7.0 (m, 1H), 3.2-3.1 (m, 2H), 3.0-2.7 (m, 4H) , 1.8-1.6 (m, 4H), 1.4 (s, 9H). - / 26345 PCT /US94/03223 N— <BOC) -R—ami nomethvl-2—naphthoic acid (14) The remaining naphthoate (11) was treated with 2,3-dichloro-5,6-dicyano-l,4-benzoquinone (DDQ) in dioxane 5 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-aminoxnethyl-2-naphthoic acid (14) as 10 an intermediate for incorporation into the cyclic peptide.

Part A - A solution of methyl 8-cyano-5,6-dihydro-2-15 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 120°C over 16 hours. The reaction mixture was poured into ice water and extracted with ethyl acetate. The combined organic layers were dried 20 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 = 178-182°C; ^-H NMR (CDCI3) d 8.95 (s, 25 1H), 8.3-8.2 (m, 1H)f 8.15-8.10 (m, 1H), 8.0-7.95 (m, 2H), 7.7-7.6 (m, IK), 4.05 (s, 1H).

Part B - A mixture of methyl 8-cyano-2-naphthoate (1.0 g, 0.0047 mol) in methanol (35 mL) with concentrated 30 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 Celite® and washed with methanol. The filtrate was evaporated to 35 dryness under reduced pressure and the residue was triturated with hexane to give methyl 8-aminomethyl-2- 263 45 6 naphthoate (0-76 g, 0.0035 mol, 75%) as an oil. NMR (DMSO) d 8.75 (s, 1H), 8.5 (bs, 2H), 8.2-8.05 (m, 3H), 7.75-7.70 (m, 2H), 4.6 (s, 2H), 3.95 (m, 3H).

Part C - To a solution of methyl 8-aminomethyl-2-naphthoate (0.75 g, 0.0035 mol) in dry tetrahydrofuran (50 mL), cooled to 0°C, 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 mol, 95%) as a white solid, mp «= 223-225°C; 1H NMR (DMSO) d 8.6 (8, 1H), 8.1-7.9 (m, 1H) , 7.8-7.7 (m, 4H), 7.55-7.5 (m, 1H), 7,45-7.35 (m, 2H), 4.2 (s, 2H).

Part D - A solution of 8-aminomethyl-2-naphthoic acid (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 (IN) 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 - 190-191°C; NMR (DMSO) d 13.1 (bs, 1H), 8.8 (s, 1H), 8.0 (q, 2H, J-7.9 Hz), 7.9 (d, 1H, J-8,1 Hz), 7.6 (t, 1H, J=7.5 Hz), 7.65-7.55 (m, 2H), 4.6 (d, 2H, J»5^5 Hz), 1.4 (s, 9H).

RECEIVE PCT /US94/03223 263456 Synthpsis of Cvrlic Peptides 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 10 substitution level of the resin), 1 equivalent of HBTU, and 3 equivalent of NMM. Alternatively, Boc-Mamb (1 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 15 substitution level is then determined using either the picric acid test (Sarin, Kent, Tarn, and Merrifield, (1981) Anal. Biochem. 117, 145-157) or the quantitative ninhydrin assay (Gisin (1972) Anal. Chim. Acts. 58, 248-24 9). Unreacted oxime groups are blocked using 0.5 M 20 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 25 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 30 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. 35 After the linear peptide had been assembled, the N- terminal Boc group is removed by treatment with 25% TFA h,Z. PA 1**1**'' • 'if v'KX.j in DCM for 30 minutts. The resin treatment with 10% DIEA in DCM. d by concomitant cleavage of the peptide is accomplished using the method of Osapay and Taylor ((1990) J. Am.

Chem. Soc., ■ 112, 6046) by suspending the resin in approximately 10 ml/g of DMF, aidding 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 10 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 /n-cresol or anisole as scavenger at 0°C for 20 to 60 minutes to remove side chain 20 protecting.groups. The crude product may be purified by reversed-phase HPLC using a 2.5 cm preparative Vydac. CI? 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(OcKex), 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, 30 Boc-D-Nle, Boc-D-Leu, Boc-D-Val, Joc-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-C1Z), Boc-^-Ala, Boc-D-Pro, Boc-D-Phe, Boc-D-Tyr(Cl2Bzl), Boc-NMe-Amf(CBZ), Boc-aminotetralin-35 carboxylic acid, Boc-aminomethylnaphthoic acid, Boc-4-aminomethylbenzoic acid, or Boc-NMeGly.

NX PATE? 1- OCT 1996 WO 94/22910 - 77- PCT/US1M/03223 263456 The synthesis of the compounds of the invention is further exemplified in PCT Patent Application International Publication Number W093/07170 (Publication Date April 15, 1993) and as set forth below. The Tables below set forth representative compounds of the present invention.

Example 3a cyclo-(Abu-NMeArg-Gly-Asp-Mamb); the compound of formula (II) wherein J - Abu, K * NMeArg, L = Gly, M «= Asp, R1 « H, R2 « H The title compound was prepared using the general 15 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 20 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 2 5 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 0.4 5%/ min. gradient of 9 to 27% acetonitrile containing 0.1% TFA and then lyophilized to give the TFA salt of 30 the title compound as a fluffy white solid (1.92% recovery, overall yield 0.574%); FAB-MS: [M+H] ■ 561.39.

Example 4 Crystallization of tha Comoound of Example 4 and t.ho.

Preparation of Salt Forms of the Compound of Example 4 r~ 263456 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 10 solution of 1.0 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 5"C in an ice bath. After stirring 1 hour, the solution was filtered and the solid 15 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 20 equivalent of the appropriate acid: Example 4 (biphenylsulfonate) : zwitterion + 1.0 equivalent biphenylsulfonic acid. o Example 4 (a-naphthalenesulfonate) : zwitterion + 1.0 equiv. a-naphthalenesulfonic acid.

Example 4 (f5-naphthalenesulfonate) : zwitterion + 1.0 equiv. {5-naphthalenesulfonic acid, Example 4 (benzenesulfonate): zwitterion + 1.0 equiv. benezene-sulfonic acid.

Example 4 (p-toluenesulfonate): zwitterion + 1.0 equiv. p-toluene-sulfonic acid.

WO 94/22910 -79- PCT/US94/03223 26 3 45 6 :d!H£5aun?rof Example The following salts of the cflfflpCTuncrof Example 4 were prepared by crystallization of the compound from aqueous systems.

Example 4 (sulfate): 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 10 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 25 volume of isopropanol, and dried under vacuum to constant weight.

Example 4 (p-toluenesulfonate): 100 mg zwitterion dissolved per ml water +1.2 molar 30 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. r WO 94/22910 _Q0_ PCT/US94/03223 263456 cyclo-(D-Met-NMeArg-Gly-Asp-Mamb); the compound of formula (II) wherein J «■ D-Met, K = 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 the attachment of Boc-Mamb to the resin. The peptide was prepared on a 0.179 mmol scale to give the protected 10 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 15 title compound (72 mg; 92.3% 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 14.4 to 23.4% acetonitrile containing 0.1% TFA and then lyophilized to 20 give the TFA salt of the title compound as a fluffy white solid (13.2% recovery, overall yield 7.4%); FAB-MS: [M+Hj - 607.3.

Example 401 cyclo-(D-Abu-NMeArg-Gly-D-Asp-Mamb) ; compound of formula (II) 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 35 on a 0.596 mmol scale to give the protected cyclic peptide (273 mg, 57.6%). The peptide (263 mg) and 0.263 1 -8i- PCT /US94/03223 263456 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.

Example 4Q2 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 20 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 0.59 6 mmol scale to give the protected cyclic 25 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 30 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 35 salt of the title compound as a fluffy white solid (2.3% recovery, overall yield 0.99%); FAB-MS: [M+H] ■» 561.36.

I OCT \ 263456 Example 4 03 Cyclo-(D-Ala-p-guanidinyl-Phe-Gly-Asp-Mamb); the compound of formula (II) wherein J » D-Ala, K * p-guanidinyl-Phe, L «■ Gly, M = Asp R^- ~ H, R2 H Dissolved 25 mg (38.3 (imoles) 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 a^id and 18.7 mg of 4-dimethyl-aminopyridine. After refiuxing for an additional 3 hours, the reacticr. 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. 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, R^- «= H, R^ = H NH O —< o 83- 263456 IM.

C > NHj-C(«NH)SOjH OMAP, EIOH Dissolved 10.53 mg (16.3 Umoles) 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.

Examplfis 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, R1 - H, R2 - H cyclo-(D-Abu-NMeLys-Gly-Asp-Mamb); the compound of formula (Hi wherein J « D-Abu, K « NMeLys, L » Gly, M « Asp, R1 - H, R2 - H WO 94/22910 - 84- - _ . _ PCT/US94/03223 263 456 Di-N-methyl amino acid derivatives may be prepared using methods which have been described previously (Olsen, J. Org. Chem. (1970) 35: 1912) or, alternatively, through the use of NaH/CH3l. The mono-5 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-10 Mamb) was obtained in 0.31% overall yield, FAB-MS: [M+H] = 54 7.3. Cyclo-(D-Abu-NMeLys-Gly-Asp-Mamb) was obtained in 0.25% overall yield, FAB-MS: [M+II] •= 533.3.

Example lQQa cvclo- (D-Abu-NMeArcr-Glv—Asp-3-aminoniethv 1-6- phlorobenzoir acid) The title compound was prepare by the general solution-phase procedure described above for cyclo-(D-20 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 25 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; NMR (Dg-DMSO) 9.00 (d, 1H), 8.4 0 (m, 2H), 7.50 (m, 1H), 7.40 30 (d, 1H), '.30 (m, 2H), 7.15 (s, 1H), 7.00 (br s, 4H), 5.15 (dd, 1H), 4.65 <q, 1H) , 4.50 (dd, 1H), 4.40 (q, 1H), 4.05 (dd, 1H), 3.95 (dd, 1H), 3.65 (dd, 1H), 3.10 (q, 2H), 3.05 (s, 3H), 2.75 (dd, 1H), 2.50 (m, 1H) , 1.95 (m, 1H), 1.75 (m, 2H), 1.60 (m, 1H), 1.35 <m, 2H), 0.95 35 (t, 3H); FAB-MS: [M+H] - 595.4.

WO 94/22910 263 456 PCT/US94/03223 Example 1QQE cvclo- (D-Ahii-NMPAry-Glv-Asp-^-aminomethvl-S-jLQdQbenzQic acid) 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 10 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; NMR (Dg-DMSO) 8.90 (d, 1H), 8.40 (m, 2H), 7.70 (d, 1H), 7.50 (m, 1H), 7.30 (m, 1H), 7.05 (s, 1H) , 7.00 (d, 1H), 7.00 (br s, 4H), 5.15 (dd, 1H), 4.65 (q, 1H) , 4.45 (dd, 1H), 4.40 (q, 1H), 4.00 (q, 1H), 3.90 (q, 1H) , 3.65 (dd, 1H), 20 3.10 (q, 2H), 3.05 (S, 3H), 2.70 (dd, 1H) , 2.50 (m, 1H), 1.95 (m, 1H), 1.75 (m, 2H), 1.60 (m, 1H) , 1.40 (m, 2H), 0.95 (t, 3H); FAB-MS: [M+H] - 687.3.

Example 100r. 2 5 cvrlo- fD-Ahn-NMgAro-fi 1 v-Asn-3-aminnmot-hvl-6- methylhfinzoic acid) (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 35 HF in the presence of anisole as scavenger. Purification was accomplished by reversed-phase HPLC on a preparative WO 94/22910 -B6_- PCT/US94/03223 263456 Vydac CI8 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; NMR 5 (D6-DMSO) 8.70 (d, 1H), 8.40 (d, 1H), 8.30 (t, 1H) , 7.50 (m, 1H), 7.45 (m, 1H), 7.15 (q, 2H), 7.05 <s, IK), 7.00 (br s, 4H) , 5.15 (dd, 1H), 4.65 (q, 1H), 4.45 (m, 2H), 4.00 (m, 2H), 3.65 (dd, 1H), 3.10 (q, 2H) , 3.05 <s, 3H) , 2.75 (dd, 1H), 2,50 (m, 1H), 2.30 (s, 3H), 2.00 <m, 1H), 10 1.75 (m, 2H), 1.60 (m, 1H), 1.35 <m, 2H), 0.95 (t, 3H); FAB-MS: [M+H] - 575.4.

Rspresentarlve Prodrugs Step 1: Nfl-benzvloxvrarbonvl-wa-mf»thv3-4-c:vano-L-2-aminobut-.vrie aelri 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 20 mmol). The solution was stirred at room temperature for 2 h ard concentrated at 30° C tc 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 25 twice. The combined organic solution was washed with brine four times, dried (MgSC>4), and concentrated. The oily product was dried over KOH overnight.

The dried oily product was taken up in 300 mL dry THF and 4 9.8 mL (900 mmol) methyl iodide in a flask 30 bott.le 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 35 added, and after stirring for 10 min, 100 mL water was added slowly. The solution was,acidified with a few 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. *H-NMR (CDC13): 5-2.15 <m, 1H); 2.38 (m, 1H); 2.42 (m, 2H); 2.96 & 2.98 (2s, cis & trans N-CH3); 4.62 (m, 1H); 4.90 (b, 1H); 5.19 (s, 2H); 7.35 (m, 5H) .

Step 2: Nfl-methvl—4—ryano-L—2-aminobutvric aeid-N- carboxyanhydrlde 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 nmiol) 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 5.8 6 g 25 (87%). mp 90-92° C. 1H-NMR (CDCI3) : -6=2.18 (m, 1H) ; 2.39 (m, 1H); 2.60 (m, 2H); 3.02 (s, 3H); 4.28 (m, 1H). step 3: N-Bpc-D-2-aminphut,yryl-Na-inethyl-4--cYanp-k-2-aminobutvrvl-glycine t.-butyl ester 30 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 raL dry acetonitrile, the solution was stirred at -20° C for 1 35 h, and the solvent was reduced to about 10 mL. "263456 PCT /US94/03223 To a solution oi 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 nunol) and the mixture was 5 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 10 was washed with NaHC03 four times and with brine three times, dried over MgS04, 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—Boc—D—2—aminobutvrvl—NB—methvl -Nflk-NfiH-(bisbenzvloxvcarbonyl)—L—aroinvl-glycine t-butvl ester 20 The compound of Step 3 (4.63 g, 10.5 mmol) was dissolved :n 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 25 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'-bisbenz/loxycarbonyl-S-methylisothiourea (3.58 g, 10 j0 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% 35 NaHC03 and brine, dried (MgS04), and concentrated.

Crystallization from ethyl ether-petroleum ether gave © 9 7.2 g (95%) product. FAB-MS (MH+): Calculated 755.4; Found 755.4.

Step 5: D-?-aminr»hnryrv1-Ng-inf>t-hvl-Nm. Nttl-5 <hlsb«=>nzylnxyrarbnnyl) —L—arainvj—glycine TFA salt.

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 10 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- raminnmot-hyl 1 benzoic ariH hvrirnrhl oririe 15 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 20 precooled concentrated HCl followed by 300 mg platinum(TV) 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 2 5 the solvent was removed under reduced pressure to give 6.2 g (82%) product. 1H-NMR (DMSO-dg): 6-4.08 (d, 2H); 7.53 (t, 1H); 7.80 (d, 1H); 7.94 (d, 1H); 8.10 (s, 1H); 8.65 (s, 3H).

Step 7: Fmnc-I-ascartvl(t-butvl)-3-famlnomethvl)- benzoic acid To a solution of FmocAsptBu*-)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) 35 diisopropylethylamine, and after stirring at room temperature for 5 h, 200 mL 5% citric acid was added and I WO 94/22910 n ^ ^ PCT/US94/03223 •263456 the solution was extracted with ethyl acetate twice. The combined extracts were washed with brine, dried (MgSO^) , and concentrated to give a solid which was washed with ether-petroleum ether and dried. Yield 16.3 g (100%). 1H-NMR (DMSO-d6): 8=1.35 (s, 8H); 2.48 (dd, 1H) ; 2.70 (dd, 1H); 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, 1H); 12.92 (s, 1H).

Step 8: Fmor-L—aspartvl(fc-butvl)-3-10 (aminomethyl) benznvl —D-2-wmi nobutvrvl-N£-methvl—NM. Nttl-(bisbenzvloxvcarbonvli-r,-arainvl-glvcine 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 15 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 2C 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 (MgS04) , and concentrated. The 25 residue was triturated with ether-petroleum ether to give 22 g (98%) product. FAB-MS (MH+): Calculated 1125.5; Found 1125.7.

Step 9 : Cvclo rX-afiparttyl (t-butvl)-3-30 (aminomethyl 1 benzoyl—D-?—aminobutvrvl-Mtt. Nttl-(bisbenzvloxvcarbonvl) -L-arcrlnvl-crlvcvl 1 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 35 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. 263456 Ethyl acetate (1000 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 5 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: Cvclo fX.-aspartvl-3- (aminomethyl) benzoyl-P-2-10 aminobutvrvl-NK. ml- (blsbenzvloxvearbonvll-I-arainvl- qlvcyll 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 15 concentrated. The residue was triturated with ether to give 14.5 g (97%) product. FAB-MS (MH+) : Calculated 82 9.4; Found 829.1.

Example 301 CvclofL-aspartyl (acet.p;xyn\ethyl}-3-{arnxnpniethyl)benzpyl- P-2-aminobutvrvl-L-aroinvl-alvcvn (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 (MgS04), 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) 35 pure product. FAB-MS (MH+): Calculated 633.3; Found A mixture containing the compound of Step 10 633.2. f 263456 F.xamole 308 Cvclo fL-aspartvl(pivalovloxvmethvl)-3- (aminomethyl) bgnzny T-D-2-aminobutvrvl-.L-arqinvl-alvcvl 1 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), Nal (4.5 g, 30 mmol) in 10 mL DMF was stirred at room temperature 10 for 18 h. Ethyl acetate (100 mL) was added and the solution was washed with brine 3 times, dried (MgS04), 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 15 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 20 water. Purification using semipreparative HPLC gave 1 g (26%) pure product. FAB-MS (MH4): Calculated 675.3; Found 675.3.

Example 351 CvclofL-aspartvI-(isopropvloxvcarbonvl-oxymethvlt-3- aminomethvl)benzovl-P-2-aminobutvrvl-L-aroinvl-alvcvl1 A mixture containing the compound of Step 10 (4.14 g, 5 mmol), chloromethyl isopropyl carbonate (4.58 g, 30 30 mmol), triethylamine (2.8 mL, 20 mmol), Nal (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 (MgS04), 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 ~93" W/US94/03223 263 4 5 o 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 5 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 10 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 15 Attorney Docket No. DM-6665, named inventors De Grado, Dorow, Ward, and Xue.

Dtilit.v 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 25 compound is considered to be active in these assays if it has an IC50 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.

Platelet Aggregation Assay; 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 35 citrated Vacutainer tubes. The blood was centrifuged for 15 minutes at 150 x g at room temperature, and N.z. patpnt office Hf'T 19% 7r_Q PCT/USS4/33223 ,26)3^r5o£ platelet-rich plasma/££E^r)^aslraiflo'<\ra. 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 5 Aggregation Profiler), using PPP as the blank (100% transmittance) . 200 Jil of PRP was added to each micro test tube, and transmittance was set to 0%. 20 fil of various agonists (ADP, collagen, arachidonate, epinephrine, thrombin) were added to each tube, and the 10 aggregation profiles were plotted (% transmittance versus time). The results were expressed as % inhibition of agonist-induced platelet aggregation. For the IC50 evaluation, the test compounds were added at various concentrations prior to the activation of the 15 platelets.

PlatfilPf-TMhr-Snnaen Binding Assay; Binding of ^•25i_fibrinogen to platelets was performed as described by Bennett et al. (1983) Proc. Natl. Acad. Sci. USA 80: 20 2417-2422, with some modifications as described below. Human PRP (^-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 25 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, *25j_fibrinogen. The bound to the activated, platelets was separated from the free form by centrifugation and then counted on a gamma counter. For an IC50 evaluation, the test compounds were added at various concentrations prior to the activation of the platelets.

WO 94/22910 PCT/US94/03223 -9 5- ~~ 263456 The novel cyclic glycoprotein Ilb/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 5 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 IC50 value (that is, the molar concentration of the cyclic 10 compound capable of achieving 50% clot lysis) of less than about 1 mM, more preferably an IC50 value of less than about 0.1 mM, even more preferably an IC50 value of less than about 0.01 mM, still more preferably an IC50 value of less than about 0.001 mM, and most preferably 15 an IC50 value of about 0.0005 mM.

IC50 determinations may be made using a standard thrombolysis assay, as described below. Another class of preferred thrombolytic compounds of the invention 20 include those compounds which have a Kd of < 100 nM, preferably < 10 nM, most preferably 0.1 to 1.0 nM.

Thrombolytic Assav: 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 25 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, 30 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 35 poor plasma (PPP), which served as a plasminogen source.

N.Z. PATBNT_OfflC€ 1- ar.T VWB RECElVtD WO 94/22910 PCT/US94/03223 -96-. 26 3 45 6 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 5 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 10 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 15 at various concentrations, and the percentage of lysis caused by the test compounds was calculated.

Platelftf. Granular SerrPt-.ion Studies. The role of the claimed platelet GPIIb/IIIa receptor antagonists on 20 the modulation of platelet granular secretion from the a-granules, dense granules or intracellular Ca+2 binding proteins was examined. This class of compounds did not have any significant effect on platelet granular secretion of plasminogen activator inhibitor type-1 25 (PAI-1) from a-granules, the mobilization of intracellular calcium stores or the secretion of the vasoconstrictor serotonin from the denos: granules. However, other antiplatelet agents such as aspirin or the antithrombin hirudin has been shown to inhibit 30 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. '> ,.

OCT 19% KEOEiVED wo ,4,22910 PCT/US94/03223 263456 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 5 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, 10 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 15 of this invention, and combination products containing the same, will be readily apparent from the disclosures herein.

Platelet GPIIb/IIIa Binding Affinity In the human gel-purified platelet (h-GPP) 125I-fibrinogen binding assay, representative compounds of the present invention demonstrated high affinity in inhibiting the 125ibrinogen binding to h-GPP (IC50 ■= - 100 nM) regardless of the agonist used. In an 25 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 IC50 of 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. .98. 2b 3 4 5 "0—03223 Compound A (Example 3) was shown to displace 125j_fibrinogen bound to activated platelets . In this study, fibrinogen bound to activated platelets was incubated for 20 minutes prior to the addition of 5 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 ^H-labeled 10 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 Kj_ of 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 20 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 25 (Fig. Ia) .

The effect of Compound A on the lysis of a pre-formed platelet-rich clot was also examined. In this regard, the thrombolytic efficacy of Compound A was also evaluated (Fig. Ib and II). Compounds A and B 30 (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 efficacy with standard thrombolytics in lysing a platelet-rich thrombus (Fig. Illb). A 35 concentration-dependent lytic effect with an IC50 of 0.5-1.0 uM for compounds A and B was shown (Fig. Ib). ^ i PCT /US94/Q3223 "■ 263 45 6 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, 5 urokinase or t-PA in lysing a pre-formed platelet-rich clot (Fig. Ill). These results suggest an in vivo lytic potential for disclosed compounds of the present invention. Additionally, administration of these novel antagonists is expected to significantly reduce the 10 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 15 closure. Hence, the disclosed analogs might be an effective adjunct to thrombolytic therapy or angioplasty.

The cyclic glycoprotein Ilb/III antagonist 2 0 compounds of this invention have also been shown to displace l25I-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 2 5 affinity in displacing fibrinogen from an already formed platelet-ricn 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 30 preceding examples.

Figure 1 Representative cyclic compounds of the present 35 invention, namely the compound of Example 3 (cyclo-(D-Abu-NMeArg-Gly-Asp-Mamb; the compound of formula (II) 26343d wherein R1 and R2 are H, J is D-2-aminobutyric acid, K is a-N-methylarginine, 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 5 compound of formula (II) wherein R1 and are Hf 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 la). Figure 1 shows the effect of 0.1 uM Compound or 10 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 15 inhibit platelet aggregation induced by agonists, but also to deaggregate platelets after the initiation of aggregation (Figure la). The deaggregation efficacy of compound A was dependent on its concentration and the time of addition post-initiation of platelet activation. 20 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 25 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 30 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 35 A and B) demonstrated a significant effect on the lysis of an already formed platelet-rich clot. As the results WO 94/22910 PCT/US94/03223 101 263 45 6 indicated, Compounds A and B had IC50 values of about 0.5-1.0 uM. By comparison, the RGDS linear peptide was much less effective, even at substantially higher concentrations (IC50 > 1 mM).

Figure 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 10~3 H concentration of ADP. As a comparison, the standard thrombolytics tissue plasminogen activator (tPA; 10 ng/ml), urokinase (UK; 900 units/ml) and streptokinase (SK; 500 units/ml) were also tested in the thrombolytic assay. The results are shown in Figure II. 15 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 20 Compound B providing significantly better clot lysis than streptokinase. As the results indicated, Compound A had an excellent lysis percentage of 70% or greater.

Figure III Figure III shows the effect of 1 uM of Compound A 25 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 Jlg/ml) , urokinase (UK; 900 units/ml) and streptokinase (SK; 500 units/ml). A synergy (greater than additive effect between the standard thrombolytics WO 94/22910 PCT/US94/03223 "" 26^56 and the Ilb/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 5 using the thrombolytic assay described above, both alone and in combination with the standard thrombolytics tissue plasminogen activator (tPA; 10 jlg/ml) , urokinase (UK; 900 units/ml) and streptokinase (SK; 500 units/ml). As the results indicate, the combination of Compound C 10 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 15 using the thrombolytic assay described above, both alone and in combination with the standard thrombolytics tissue plasminogen activator (tPA; 10 fig/ml) , urokinase (UK; 900 units/ml) and streptokinase (SK; 500 units/ml). As the results indicate, the combination of Compound D 20 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. 30 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 WO 94/22910 -10 3- PCT/US94/03223 263456 standard thrombolytics and the Ilb/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 10 femoral artery. Additionally, Compound A in combination with sub-optimum doses of the standard thrombolytic, streptokinase (75 KO) resulted in a significant synergistic effect in fully lysing the thrombus with subsequent restoration of flow, and the prevention of 15 reocclusion. Data represent mean ± SEM. n=3-6 in each group.

Figure VIII Effects of Compound A vs saline, when given to 2 0 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 2 5 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.

Antiplatelet, combination of the cyclic GPIIb/IIIa receptor antagonist of the present invention and aspirin and/or heparin.

Methods: Twelve purpose bred mongrel dogs (8-15 months of age) of either sex weighing between 8-12 kg were WO 94/22910 -10 4 - PCT/US94/03223 263456 anesthetized with thiamylal sodium (15 mg/kg, i.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 5 dissected and cannulated for &rterial blood pressure and heart rate monitoring, blood sampling, and intravenous injections. ment 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.

Treati (a) Serial blood samples were withdrawn for ex vivo 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 35 with the agent's site of action, glycoprotein Ilb/IIIa (GPIIb/IIIa), in the body of a mammal. They can be WO 94/22910 iril- PCT/US94/03223 — JLU3- 263456 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 5 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, 10 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 15 selected on the basis of the chosen route of administration and standard pharmaceutical practice.

The dosage of the novel cyclic compounds of this invention administered will, of course, vary depending upon known factors, such as the pharmacodynamic 20 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 25 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 30 pharmaceutical 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 35 powders, or in liquid dosage forms, such as elixirs, WO 94/22910 106 PCT/US94/03223 263456 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 5 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 10 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 15 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 20 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 25 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 30 propyl-paraben, and chlorobutanol.

Suitable pharmaceutical carriers are described in Remington's Pharmaceutical Sciences. Mack Publishing Company, a standard reference text in this field.

Useful pharmaceutical dosage-forms for 35 administration of the compounds of this invention can be illustrated as follows: -10 8 - PCT/US94/03223 26345 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 5 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 Ilb/IIIa compounds of this invention and the anti-coagulant agent, anti-platelet 10 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, 15 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 2 0 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 25 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 Ilb/IIIa antagonist compounds of this invention and the 30 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 35 component of the combination product may be administered orally, and another component may be administered WO 94/22910 -107- PCT/US94/03223 263456 Capsules A large number of unit capsules are prepared by filling standard two-piece hard gelatin capsules each 5 with 100 milligrams of powdered active ingredient, 150 milligrams of lactose, 50 milligrams of cellulose, and 6 milligrams magnesium stearate.

Soft Gelatin Capsules 10 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 15 ingredient. The capsules are washed and dried.

Tablets A large number of tablets are prepared by conventional procedures so that the dosage unit was 100 20 milligrams of active ingredient, 0.2 milligrams of colloidal silicon dioxide, 5 milligrams of magnesium stearate, 275 milligrams of microcrystalline ce-llulose, 11 milligrams of starch and 98.8 milligrams of lactose. Appropriate coatings may be applied to increase 25 palatability or delay absorption.

The combination products of this invention, such as the novel cyclic Ilb/IIIa antagonist compounds of this invention in combination with an anti-coagulant agent 30 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 35 combinations thereof, can be in any dosage form, such as r intravenously) . The dosage of the^c^i^tXpn^rfWIucts of the invention may vary dependin^"uponMfarious 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 15 dosage may be about 0.01 to 10 milligrams of the cyclic compound of this invention and about 1 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 20 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 25 about 1 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 30 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 35 typical dosage form of this type of combination product, such as a tablet, the yiovel compounds of this invention WO 94/22910 PCT/US94/03223 26 3 45 6 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 5 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 10 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 15 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 20 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 25 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 30 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 35 warfarin or heparin, or a novel compound of this invention and an anti-platelet agent such as aspirin, joWHNT OFFIS j 1- OCT 1996 —■ ' i-0 263456 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 5 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 10 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 15 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 20 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 25 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 30 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 35 release polymer, and the other component is also coated with a polymer such as a lowviscosity grade of WO 94/22910 -112- PCT/US94/03223 263456 hydroxypropyl 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 5 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 10 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, 15 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 20 form of a plurality of microtablets, particles, granules or non-peril/, 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 25 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 30 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 35 inhibition of platelet aggregation, the treatment of blood clots, and/or-the treatment of thromboembolic WO 94/22910 -113 - S'iT'SiS94'] disorders, which comprise a therapeutically effective amount of a novel cyclic platelet glycoprotein Ilb/IIIa compound of this invention along with a therapeutically effective amount of an anti-coagulant agent such as 5 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 10 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 15 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-20 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, 25 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 30 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 IC50 value in the platelet aggregation aasay described above. The IC50 values are expressed as: +++ = IC50 value of less than 1 uM; ++ » IC50 value of 1 uM to 10 uM; and ; + » IC50 value of greater than 10 uM to about 5 1Q0 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.

Tahlp 1 263^56 Gly R4 0 I II -N-C-C- R3 R5 The optical isomer of J is indicated unless otherwise indicated.

B? JB4 B5 RiO-H FAB-MS Optical MP,..

(M+H) Isomer l 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) 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)5CH3 (D) 8a H H (CH2)5CH3 (L) N.Z. PATENT OFFICE 1- OCT 1996 RECEIVED 263456 tahlfi 1 (sont.inued) ELx.. 9 £3 H H E5 (CH2)7CH3 FAB-MS (M+H).

Optical Isomer (D) 9a H H (CH2)7CH3 (L) H H C(CH3)3 (D) 10a H H C(CH3)3 (L) 11 H H phenyl 609.27 (D) 11a H H phenyl (L) 12 H H phenylmethyl 623.28 (D) 12a H H phenylmethyl (L) 13 H H CH20H (D) 13a H H CH20H (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 K (CH2)4NH2 601.32 <D) 13g H H (CH2)4NH2 (L) 13h H H <CH2)4NHC<-NH)NH2 (D) 13i H H (CH2)4NHC(»NH)NH2 (L) 13 j H H (ch2)5NH2 <D) 13k H H (CH2)5NH2 (L) 131 H H <CH2)5NHC(»NH)NH2 (D) 13m H H (CH2)5NHC(-NH)NH2 (L) I3n H H (CH2)4ch3 (D) 13o H H (CH2)4CH3 <L) WO 94/22910 PCT/US94/03223 263456 TahTg 1. /continued) Ex, Hsu. 13p £3 H R4 H B5 (CH2)6ch3 E&BzMS 1H+H) Optical la.oroer. (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 K H CH2SH (L) 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 (1) 18 CH3 H H 54"7.34 +++ 13 H CH3 CH3 - H CH2 CH3 ch2ch3 21 H H cyclopentyl (D) 21a H H cyclopentyl <L) 22 H H cyclohexyl (D) 22a H H cyclohexyl (L) +++ 23 K H cyclohexylmethyl (D) 23c H H cyclohexylmethyl (L) 23a H H CH(CH3)2 Rl0-I 701.37 (D) 23b H H CH(cr3)2 R10-l (L) 23d H H 23e H H 23f H H 23g H H 23h H 23 j H H H H O H O NHaH I I I I (CHa)4 — N —C —C—C —|| H H H O I II .(C^)« —N—C 263 45 6 HO o I 8 «=\ II — (CH2>4 — N—C — H — (CHj)4 — N—3»0 n N(CHj)] +++ +++ +++ +++ +++ 23k H 231 H H H 23m H H H O H H o CH, I II I I II I — (CHj), — N—C—C —N —C-O -C—CHj H CHj - (CHa)4 - O K H II I I ■C—C—N I H -H H O H H 0 1 H I I (CHj)4 -N—C—C-N +++ +++ 23n H H H O II -(CH,),-H-C-0-0 +++ IfTZfVkTEMT OF riot 1~ on 1996 PCT AJS94/03223 23o H H 263456 +++ - (CHa) 23p H H - (C^)4 I H O C —0 0 1 H O c=0 1 11 -n-c-q ++ r WO 94/229JO PCT AJS94/03223 263456 Table. .3 (continued) y NMeArg / J \ o=c Gly v Asp \ NH Sat..

No. 24 FftB-MS (MfSl Optical iCifl Isomer 573.46 (L) 573.35 (D) +++ 26 "nh2^ 27 28 -nh^JL O 28a <D) N.Z. PATENT OfFICC 1- OCT 1996 RECEIVED WO 94/229!0 EX. jI lie. 28b 28c 28d 28e Tflhlft \ front.) FftB-MS (M+S) PCT /US94/03223 263456 547.28 Optical !£££ Isomer (L) (D) (L) +++ Ex• E3 B4 No . 28f H H 28g H H B- -CH ^ // OH OH F-AB-M5 Optical (M+s^ Isomer 639.54 (D) (L) +++ Specifically provided by the present invention are those compounds of Table 1 wherein Asp is replaced by a residue selected from: OtMeAsp; (JMeAsp; NMeAsp; D-Asp; Asp-(methyicarbonyloxymethyl ester); Asp-(ethylcarbonyloxymethyl ester); Asp-(t-butylcarbonyloxymethyl ester) ; Asp-(cyclohexylcarbonyloxymethyi ester); WO 94/22910 PCT/US94/03223 263456 Asp-(1-(methylcarbonyloxy)ethyl ester); Asp- (1-(ethylcarbonyloxy)ethyl ester); Asp-(1-(t-butylcarbonyloxy)ethyl ester); Asp-(1-(cyclohexylcarbonyloxy)ethyl ester); 5 Asp-(i-propyloxycarbonyloxymethyl ester); Asp-(cyclohexylcarbonyloxymethyi ester); Asp-(t-butyloxycarbonyloxymethyl ester); Asp-(1-(i-propyloxycarbonyloxy)ethyl ester); Asp-(1-(cyclohexyloxycarbonyloxy)ethyl ester); 10 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-(l-(2-(2-methoxypropyl)carbonyloxy)ethyl ester). f N.Z. PAI^MTOfnCEj 1- QCT19SB | 26345 Table 2 R O I B -n-c-c-I I J m « R5 The optical isomer of J is indicated.

BlU.

B4 B5 FAB-MS.

Optical ma-.

(M+H) I acme x 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) h ch2oh 549.31 (d) ++ 35a h ch20h (l) 36 h phenylmethyl 609.25 (d) + 37 h phenylmethyl 609.26 (l) + r WO 94/22910 PCT/US94/03223 lafele,3 263 45 k - -N(R6)CH<R7)C(-O)- The optical isomer of K is indicated.

E2^ £6 e7 FAR-MR Optical !£££ Nn. (M-f Isomer 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-^~y-CH2NH2 (L> 393 H -CH2-^~y-CH2NH2 <D) 40 CH3 -CH2-£yCH2NH2 595 (L) 40a CH3 -CH2-^y-CH2NH2 (D) 41 CH3 NH (L) -CH2-^^-CH2NH-^ Table.3 (continued) 41a ch3 NH 4 5 ch3 45a CH3 -CH2-^V-CH2NH \s=s/ > nh2 42 ch3 -CH2SCH2CH2NH2 42a CH3 -CH2SCH2CH2NH2 43 CH3 -ch2sch2ch2nhc(-NH)(NH2> 43a ch3 -ch2sch2ch2nkc(-NH)(NH2) 44 ch3 -ch2ch2sch2ch2nh2 4 4a ch3 -ch2ch2sch2ch2nh2 ch2ch2sch2ch2nhc(«NH)(NH 2) CH2CH2SCH2CH2NHC(«NH)(NH 2> NH 4 6 CH3 XNH2 NH 4 6a CH3 (/ \ 'Nlfe -ch>-gk =\ f=\ ^,NH — CH,—' *—" 47 CH3 4 7a CH3 48 CH3 ~ch2 o CHjNH; 48b CH3 ""ch2 -o CH2NH? PCT /US94/03223 263456 (D) <L) (D) (L) (D) (L) (D) (L) (D> (L) (D) <L) <D) (L) (D) WO 94/22910 PCT/US94/03223 Tablft 3 (continued) -HH-C N«H, NH 49a ch3 -CH2-/ vch2nh wm/ Ex.

HC- z FAB-MS , 52 ~"^NH NH 2 53 ^m<m2 263456 £*- E6 E7 FAB-MS Opting <M+H> ISQIBftE ••• ~ --ex /"""V >NH ^ 4 9 ch3 -CH2-/ )~ch2nh *^KH2 <D) Table A 263456 & FAB-MS f M+H} Optical ic,an NO.

Isomer 4 -NHCH2C(=0)- 575. 45 +++ 54 -nhch2ch2c(-0)- 589. 32 ++ 55 -0CH2C(-0)- 56 -och2ch2c(-0)- 57 -SCH2C(-0)- 58 -sch2ch2c(=0)- 58c -NHCH(CH3)C{"0)- 589. 31 (L) + Table 5 263 45 Gly N = -N <R10)C(R8) (R9)C( =0) - ZXUL.

No.

Ee E9 B10 FAB-MS (MtH) Optical Xaomei ■I£so 4 -CH2COOH H H 575.45 (L) +++ 63 -CH2COOH CH3 H 589.29 isomer 1 63a -CH2COOH CH3 H 589.27 isomer 2 ♦ 64 -CH(CH3)COOH H H 569.43 isomer 1 ♦ +4 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 [NTZ. PATHHT CrgCE i - or,i Wb 263456 The optical isomer of -CH(R^)N(R^)- is indicated.

Ex . No . e1 E2 fab- Optical msfm+hi Isomer 4 h h 575.45 +++ 68 ch3 h 589.31 isomer 1 +++ 68a ch3 h 58 9.31 isomer 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 71a 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 Exam R1 Etis.

Ita- Tahlft 6 (ronr E2 263456 fab- Optical MS(M+H) isomer 76 phenyl H 651. 33 isomer 76a phenyl • H 651. 33 isomer 77 cyclopentyl H R 77a cyclopentyl H S 78 cyclohexyl H R 78a cyclohexyl H S 79 H CH3 589 .33 80 H CH2CH3 81 H CH2CH2CH3 82 H CH(CH3)2 83 / CH2CH2CH2CH3 84 H C (ch3) 3 85 H CH(CH3)CH2CH3 86 H benzyl ++ ++ N.Z, PATENT OFFICE 1 - OCT 1336 3 Hc.CV PCT AJS94/03223 Tflhlft 7 Ex . No . Stricture 87 NMeArg ^ D-VBI \ Asp isomer 1 88 ^NMeArg ^ Q(y D-Val \ Asp isomer 2 89a ^G1y\ NMeArg Asp I ' D-val NH "^06 isomer 1 89b ^Gly^ NMeArg Asp 89c Asp NMeArg ! D-Val 263456 FAB-MIS I&SSL (M+H) 575.41 +++ 575.44 +++ 615.34 +++ 615.35 +++ 625.32 ++ Tablet fl 263456 Strufitnrr FAB-MS fM+Hl No. 8 9d 687.33 +++ 89e 90 91 ^ <5iy. dl-NM.Orn .

I I D-Abu NH Asp 533.34 ++ 589 +++ 575 +++ WO 94/22910 -133- PCT/US94/03223 263 45 N-M«Aro **Ajp 92 / \ 575 +++ D-V«l NH nv.-~ & / J \ o=c.

K 134- Table 9 M 263456 \ NR I x CHR ,10a (VII) wherein j . D-Val, K = NMeArg, L « Gly, H ■* Asp Example glOa a10 FAB-MS XSLso Number (M+H) 93 CI M 609 +++ 94 I H 701.37 +++ 95 MeO H 623(+H20) 96 Me H 589 +++ 97 H CI 609 +++ 98 H I 701 99 H MeO 605 +++ 100 H Me 589 +++ wherein J = D-Abu, K = NMeArg, L <= Gly, M = Asp 100a H CI 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, |J-Ala, Pro, Phe, NMeGly, D-Nle, D-Phg, D-Ile, D-Phe, D-Tyr, Ala.

TCt/US94'03Z23 263*56 labia.... lfl / SR2 I °-=v^/CHRl wherein L «• Gly, M » Asp, R2 and R1 ■» H, K = -N(R6)CH(R75C(-0>-J « D-Val Ex. £6 Opt i.ca I ICso Xasmer 1 L EQ- 101 CH3 -(CHaV D rO" rO 102 CK3 -(CtfaV L 103 «3 — — (CH2)3 rO D NH l04 CH3 -(pVW _T"\h l0s c»3 _(CH2)«— D 106 CB3 -(CH2)a .-o perfuse**3233 101 CH3 108 CH3 109 cR3 „ 263456 nJ \*H —ch2o -\^/ D • -gh2o-\ / rO"' 110 -CH2OCHa- _ D -O" CH3 -CHaO^2 -o 112 CH3 -GH20(ch2)2 I _ , /snh ll3 CHS _(CH2)20 -\_J D »' " -,cW,o-q" .-O" CH3 -(CH2>2OCH2" -o 116 CB3 _(CH2)2OCtV 117 CH3 118 CH3 119 CH3 120 CH3 121 CH3 122 CH3 123 CH3 124 ch3 125 ch3 126 CH3 -ch*-0« ' 263456 D /~\ ~W ""CH2SCH2—( NH / v ~CH2SCH2—( NH -ch2s(ch2)2^Qnh -ch2s(ch2)2hQnh -o \ / —(CH2)2S—( NH ~o "(CH^S—( NH -{CH2)2SCH2—/ NH -o \ / -(CH2)2SCH2--^^NH 127 ch3 -ch2-S-* (CH2) 3-NH-CH3 L 128 CH3 -CH2-S-(CH2)3-NH-CH3 D CO K> je» »-> o U> \a o tc to o w u> n X to o tc Co o x to o a u> o EC U) O O ■—■ x DM X I u> O U» 0 x fO to 1 K tc I o o x a to to Si o JL » O U> x to •w' U> I a x • n o X X *± 10 O I as o ^ 1 o » o to x to to I a 3: 1 o o x tc — N> O I tc o u> t to a: 1 o o o x x — K> O I tc o o> l o o tc x to to O I X Sj> to to O to O tc tc 10 K> CO ( 2! EC I n x M CO 1 a ac t o o tc tc K> to o 1 tc o to I 0 tc to t*> 1 a tc t 4 t-» H M M u> U) to to to 00 <T> tn 0 0 O O 0 X X X X X to to to to to M to tO O tc u> to to o tc to to n SC u> to o n tc u> M K> vo O X O 0 0 » 0 1 X X ~ 0 — 0 to to 0 tc 0 X 1 J Xto JCM O O W 1 to t 1 1 w to 1 to t O O —s. *-v X X 0 0 to to X X to to to u> «-»• 1 » to to a a 1 1 X X 1 a a 1 X X 0 0 1 1 X X to to 0 f 0 «r« 0 I as n to x O M X I to 01 1 I o — x n Co sc M to < a x 1 o J a o to 3S n to x 1 to Cft t 1 o -» x o u> x to to ai w 1 O I a o — EC oto X I U) (ft —■ t to ~ 0 X to (a} 1 a x » o » as o — w o to x 1 U) t/3 *-* I to — 0 x to 0J « a x 1 o » x o ro x O to X I w V) 1 *•••» 0 X ro CO 1 a x » o 1 x o to X n 10 x » U> (ft 1 o w rO U) > a x » t t-1 to C3 I ro o> 01 Q a Ifl 9 en o> WO 94/22910 PCT/US94/03223 14 6 CH3 CH2-0-(CH2)3-NH- D C(CH3)3 14 7 ch3 -ch2-S-(ch2)2-NH-ch3 L 14 8 ch3 -ch2-S-(ch2)2-NH-ch3 D 14 9 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 6 ch3 -ch2-S-(ch2)2-NH- D C(CH3)3 157 ch3 ch2-0-(ch2)2-NH-ch3 L 158 ch3 ch2-0-(ch2)2-NH-ch3 D 9 ch3 ch2-0-(ch2)2-nh- l ch2ch3 160 CH3 CH2-0-(CH2)2-NH- D ch2ch3 161 ch3 ch2-0-(ch2)2-NH- L CH(ch3)2 162 CH3 CH2-0-(CH2)2-NH- D ch(ch3)2 163 CH3 CH2-0-(CH2>2-NH- L CH2CH2CH3 263 456 164 165 166 167 168 169 170 171 172 173 174 175 176 177 178 179 180 181 ch3 ch2-0-(ch2)2-NH- D ch2ch2ch3 ch3 ch2-0-(ch2)2-NH- L C (ch3) 3 ch3 ch2-0-(ch2)2~NH- D C(CH3)3 ch3 -ch2-S-(ch2)4-NH-CH3 L ch3 -ch2-S-(ch2)4-NH-ch3 D ch3 -ch2-S-(ch2)4-NH- L ch2ch3 CH3 -ch2-S-(ch2)4-NH- D ch2ch3 ch3 -ch2-S-(ch2)4-NH- L CH(CH3)2 ch3 -ch2-S-(ch2)4-NH- D CH(CH3)2 CH3 -ch2~S-(ch2)4-NH- L ch2ch2-ch3 ch3 -ch2-S-(ch2)4-NH- D ch2ch2-ch3 ch3 -ch2-S- (ch2)4-NH- L C(CH3)3 CH3 -CH2-S- (CH2)4-NH- D C(CH3)3 CH3 ch2-0-(ch2)4-NH-ch3 L CH3 ch2-0-(ch2)4~NH-ch3 D ch3 ch2-0-(ch2)4-NH- L ch2ch3 ch3 ch2-0-- (CH2) 4-NH- D ch2ch3 CH3 ch2-0-(ch2)4~NH- L CH(ch3)2 263 45 IN,Z, PATENT OFFICE 1 - OCT 1936 I 182 183 184 185 186 137 188 189 190 191 192 193 194 195 196 197 198 ch3 ch2-0-(ch2)4-nh-ch(ch3)2 ch3 ch2-0-(ch2)4-nh-ch2ch2ch3 ch3 ch2-0-(ch2)4-nh-ch2ch2ch3 ch3 ch2-0-(ch2)4-nh-c(ch3)3 ch3 ch2-0-(ch2)4-nh-c(ch3)3 ch3 -ch2-s-(ch2)5~nh-ch3 ch3 -ch2-s-(ch2)5-nh-ch3 ch3 -ch2-s-(ch2)5-nh-ch2ch3 ch3 -ch2-s-(ch2)5-nh-ch2ch3 ch3 -ch2-s-(ch2)5~nh-ch(ch3)2 ch3 -ch2-s-(ch2)5-nh-ch(ch3)2 ch3 -ch2-s-(ch2)5-nh-ch2ch2-ch3 ch3 -ch2-s-<ch2)5-nh-ch2ch2-ch3 ch3 -ch2-s-(ch2)5-nh-c(ch3)3 ch3 -ch2-s-(ch2)5-nh-c(ch3)3 ch3 ch2-0-(ch2)5-nh-ch3 ch3 ch2-o-(ch2)5-nh-ch3 ch3 ch2-o-(ch2)5-nh-ch2ch3 263 45 6 d l d l d l d l D l d l d l d l WO 94/22910 PCT/US94/03223 263 45 6 200 CH3 ch2-0-(ch2)5-NH- ch2ch3 201 CH3 CH2-0-(CH2)5-NH- CH(CH3)2 202 ch3 ch2-0-(CH2)5~NH- CH(ch3)2 203 CH3 CH2-0-(CH2)5-NH- ch2ch2ch3 204 ch3 ch2-O-(ch2)5-NH- ch2ch2ch3 205 ch3 ch2-0-(ch2)5-NH- C (ch3) 3 206 ch3 ch2-0-(ch2)5-NH- 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)2-0-(CH2)3"NH-CH3 L N.Z. PATENT 1- OCT 198 218 219 220 221 222 223 224 225 226 227 228 229 230 231 232 233 234 235 236 PCT /US94/03223 ch3 -(ch2)2-0- (ch2) 3-nh-ch3 ch3 -(ch2)2~0- (ch2) 3-nh-ch2ch3 l ch3 -(ch2)2~0- <ch2) 3-nh-ch2ch3 d ch3 -(ch2)2-0-ch(ch3)2 (ch2) 3-nh- l ch3 -<ch2)2-0-ch(ch3)2 <ch2) 3-nh- d ch3 -(ch2)2~0-ch2ch2ch3 (ch2) 3-nh- l ch3 -(ch2)2-0-ch2ch2ch3 (ch2) 3-nh- d ch3 -(ch2)2-0-c(ch3)3 <ch2) 3-nh- l ch3 -(ch2)2-0-c(ch3)3 (ch2) 3-nh- d ch3 -(ch2)2"s- (ch2) 2-nh-ch3 l ch3 -(ch2)2"s- (ch2) 2-nh-ch3 d ch3 -<ch2)2-s- (ch2) 2-kh-ch2ch3 l ch3 -(ch2)2-s- (ch2) 2-nh-ch2ch3 d ch3 - (ch2)2"s-ch(ch3)2 (ch2) 2-nh- l ch3 -<ch2)2-s-ch(ch3)2 (ch2) 2-nh- d ch3 - (ch2)2~s-ch2ch2-ch3 (ch2) 2-nh- l ch3 -(ch2)2-s-ch2ch2-ch3 (ch2) 2-nh- d ch3 - (ch2)2-s-c(ch3)3 (ch2) 2-nh- l ch3 -(ch2)2-s-c(ch3)3 (ch2) 2-nh- d 237 23.8 239 240 241 242 243 244 245 246 247 248 249 250 251 252 251 254 255 263456 CH3 -(CH2)2-0-(CH2)2-NH-CH3 L ch3 -(ch2)2-0-(ch2)2-nh-ch3 d CH3 -<CH2)2-0-(CH2)2-NH-CH2CH3 L ch3 -(ch2)2-0-(CH2)2-NH-ch2ch3 D ch3 -(ch2)2-0-(CH2)2-NH- L CH(CH3)2 CH3 -<CH2)2-0-(CH2)2-NH- D CH(CH3)2 CH3 -(ch2)2-0-(CH2)2-NH- L ch2ch2ch3 ch3 -(CH2)2-0-(CH2)2"NH- D ch2ch2ch3 CH3 -(ch2)2-0-(ch2)2-NH- L C (CH3) 3 ch3 -(ch2)2-0-(CH2)2-NH- D C (CH3) 3 ch3 -(ch2)2-S-(ch2)4-NH-ch3 L ch3 -(CH2)2-S-(ch2)4-NH-ch3 D ch3 -(ch2)2-S-(CH2)4-NH-ch2ch3 L ch3 -(CH2)2—S-(CH2)4-NH—ch2ch3 D ch3 -(CH2)2-S-(ch2)4-NH- L CH(CH3)2 ch3 -(CH2)2-S"(CH2)4-NH- D CH(ch3)2 ch3 -(ch2)2-S-(ch2)4-NH- L ch2ch2-ch3 ch3 -(ch2)2-S-(ch2)4-NH- D ch2ch2-ch3 ch3 -(ch2)2~S-(ch2)4~NH- L C(CH3)3 1 ■. —■"—"■ " 1 OCT 1996 I 256 257 258 259 260 261 262 263 264 265 266 267 2 68 269 270 271 272 273 274 275 276 277 278 145- CH3 -(CH2)2 c(ch3)3 -s- (CH2)4-nh- ch3 - (ch2)3- -nh- -CH3 CH3 - (ch2) 3' -nh- -CH3 CH3 - (ch2) 3" -nh- -CH2CH3 ch3 -(ch2)3- -nh- -ch2ch3 ch3 -<ch2)3- -nh- -ch(ch3)2 ch3 -«ch2)3- -nh- -ch(CH3)2 CH3 -<ch2)3" -nh- -CH2CH2CH3 CH3 — (ch2) 3* -nh- -CH2CH2CH3 ch3 -<ch2)3" -nh- -c (ch3) 3 CH3 - (ch2) 3* -nh- -c (ch3) 3 CH3 -(ch2)4* -nh- -CH3 CH3 -(ch2)4* -nh- -CH3 CH3 -(ch2)4- -nh- -CH2CH3 CH3 -(ch2)4- -nh- -CH2CH3 ch3 -<ch2>4- -nh- -ch(CH3)2 CH3 -(ch2)4- -nh- -ch(ch3)2 CH3 ~(ch2)4- -nh- -CH2CH2CH3 CH3 -(ch2)4- -nh- -ch2ch2ch3 ch3 ~(ch2)4- -nh- -c(ch3)3 ch3 -(ch2)4- -nh- -c (ch3) 3 ch3 -(ch2)5- -nh- -CH3 ch3 -(ch2)5-nh-CH3 CH3 -(ch2)5- -nh- •CH2CH3 ch3 -(ch2)5~ -nh- ■CH2CH3 263456 L D L D L D L D L D L D L D L D L D L D L D L d PCTAJS94/03223 263456 281 ch3 -(ch2)5-nh-ch(ch3)2 282 ch3 -(ch2)5-nh-ch(ch3)2 d 283 ch3 —(ch2)5-nh-ch2ch2ch3 l 284 ch3 -(ch2)5-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 291 ch3 -(ch2)6~nh-ch(ch3)2 l 2 92 ch3 -(ch2)6-nh-ch(ch3)2 d 2 93 ch3 -(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 10 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, f3-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 10 wherein Asp is 10 replaced by a residue selected from: OMeAsp; ^MeAsp; NMeAsp; D-Asp; Asp-(methyicarbonyloxymethyl ester); Asp-(ethylcarbonyloxymethyl ester); Asp— (t—butyl carbonyloxyiue thy 1 ester) ; Asp-(cyclohexylcarbonyloxymethyi ester); wo 94/22910 PCT/US94/03223 263456 Asp- (1-(methylcarbonyloxy)ethyl ester); Asp-(1-(ethylcarbonyloxy)ethyl ester); Asp-(1-(t-butylcarbonyloxy)ethyl ester); Asp-(1-(cyclohexylcarbonyloxy)ethyl ester); 5 Asp-(i-propyloxycarbonyloxymethyl ester)/ Asp-(cyclohexylcarbonyloxymethyi ester); Asp-(t-butyloxycarbonyloxymethyl ester); Asp-(1-(i-propyloxycarbonyloxy)ethyl ester); Asp-(1-(cyclohexyloxycarbonyloxy)ethyl ester); 10 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)-l,3-dioxa-cyclopenten-2-one-4-yDmethyl ester); Asp-( (1,3-dioxa-5-phenyl-cyclopenten-2-one-4-yl)methyl ester); Asp-(1-(2-(2-methoxypropyl)carbonyloxy)ethyl ester). % | n.z. PAir'r office] r ' — ; i ^ 'J ' EX.

Jic. 301 302 303 304 305 306 307 308 309 310 311 312 313 314 315 -148-Tablft U '263456 D-Abu B -CH2-0--CH2-0--CH2-0--CH2-0--CH2-0--CH2-0--CH2-0--CH2-0--CH2-0- -CH2-O--CH2-0--CH2-0--CH2-0--CH2-0--CH2-0- c<~0>-c{«0)' c <=0) y=o) c (=0) c(=0) •c(=0) •c<«°0) •c(=0) •c(«0) •c(=0) -c(=0) •c(=0) -c(=0) -c(»0) -ch3 -CH2CH3 -ch(ch3)2 -(CH2)2-CH3 -(ch2)3-ch3 -CH2-CH(CH3)2 -CH(CH3)-CH2-CH3 -C(CH3)3 -cyclopropyl -cyclobutyl -cyclopentyl -cyclohexyl -phenyl -4-methylphenyl -4-ethylphenyl FARMS (M+H} 633.2 675.3 ..r\ WO 94/22910 PCT/US94/03223 316 -CH2~O-C(=0)-4-isopropylphenyl 317 -CH2-0-C(-0)-4-propylphenyl 318 -CH2-0-C(=0)-4-t.-but.ylphenyl 319 -ch2-0-C(-0)-4-methoxyphenyl 320 -ch2-0-C(®0)-4-ethoxyphenyl 321 -CH2-0-C (=0) t-4-isopropyloxyphenyl 322 -CH2-0-C(-0)-4-propyloxyphenyl 323 -CH2-0-C(=0)-4-£-butoxyphenyl 324 -CH2-0-C («=0) -4-biphenyl 325 -CH(CH3)-0-C(»0)-CH3 32 6 -CH(ch3)-0-C(=0)-ch2ch3 327 -CH(CH3)-O-C(«0)-CH(CH3) 2 328 -CH(CH3)-0-C("0)-(CH2)2-CH3 32 9 -CH(CH3)-0-C(-0)-(CH2)3-CH3 330 -CH(CH3)-0-C(-0)-CH2-CH(CH3)2 331 -CH(ch3)-0-C(-0)-CH(ch3)-ch2-ch3 332 -CH (ch3) -0-C (*"0) -C (ch3) 3 333 -CH (CH3)-0-C(=0)-cyclopropyl 334 -CH(CH3)-0-C(»0)-cyclobutyl 335 -CH(ch3)-0-C(-O)-cyclopentyl 336 -CH(ch3)-0-C(-0)-cyclohexyl 337 -CH(CH3)-0-C(-0)-phenyl 338 -CH(ch3)-0-C(-0)-4-methylphenyl 33 9 -CH(ch3)-0-C(-0)-4-ethylphenyl 340 -CH(ch3)-0-C(-0)-4-isopropylphenyl 341 -CH(CH3)-0-C("O)-4-propylphenyl 263 456 342 -CH(ch3)-0-C(=0)-4-r-butylphenyl 343 -CH(ch3)-0-C(=0)-4-methoxyphenyl 344 -CH(ch3)-0-C(«0)-4-ethoxyphenyl 345 -CH (ch3) -0-C (=*0) -4-i sopropyloxyphenyl 34 6 -CH (ch3) -O-C.(-O) -4-propyloxyphenyl 347 -CH(CH3)-0-C(-»0)-4-£.-butoxyphenyl 34 8 -CH(ch3)-0-C(-0)-4-biphenyl 34 9 -ch2-O-C(-0)-O-ch3 350 -ch2-0-C(=0)-0-ch2ch3 351 -CH2-O-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 -CH2-0-C(-0)-0-CH2-CH(CH3)2 355 -CH2-O-C(=0)-0-CH(CH3)-CH2—CH3 6 -CH2-C-C(=0)-0-C(CH3)3 357 -CH2-0-C(-0)-O-cyclopropyl 358 -CH2-0-C(=0)-O-cyclobutyl 359 -CH2-0-C(«0)-O-cyclopentyl 360 -CH2-0-C(-0)-O-cyclohexyl 361 -CH2-0-C(™0)-0—phenyl 362 -CH2-0-C(=0)-0-4-methylphenyl 3 63 -CH2-0-C(=0)-0-4-ethylphenyl 3 64 -CH2-0-C(=0)-0-4-isopropylpheny1 365 -CH2-O-C(»0)-0-4-propylphenyl 366 -CH2-0-C (®*0) -0-4-t-butylphenyI '26345 PCT /US94/03223 367 -CH2-0-C(mO)-O-4-methoxyphenyl 368 -CH2-0-C(»0)-O-4-ethoxyphenyl 369 -CH2-0-C(=0)-0-4-isopropyloxyphenyl 370 -CH2-0-C(»0)-O-4-propyloxyphenyl 371 -ch2-0-C (=0) rO-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(»0)-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)-O-C<«0)-0-CH(ch3)-ch2-ch3 380 -CH(CH3)-0-C(-0)-0-C(CH3)3 381 -CH(CH3)-O-C(»0)-O-cyclopentyl 382 -CH(ch3)-O-C(=0)-O-cyclobutyl 383 -CH (CH3) -,0-C («0) -O-cyclopentyl 384 -CH(CH3)-O-C(«0)-O-cyclohexyl 385 -CH(ch3)-O-C(-0)-O-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-£,-butylphenyl p " ; . "\ 1— 26345 i::GE 391 392 393 394 395 396 397 398 399 400 401 402 403 404 -CH(CH3)-O-C(=0)-O-4-methoxyphen -CH(ch3)-O-C(-0)-O-4-ethoxyphenyl -CH(CH3)-O-C(=0)-0-4-isopropyloxyphenyl -CH(CH3)-O-C(-0)-0-4-propyloxyphenyl -CH (CH3) -O-C.(-O) -O-4-t-butoxyphenyl -CH(ch3)-O-C(«0)-0-4-bipheny1 CH2-N(CH3)2 CH2-n(CH2-CH3)2 ch2ch2—n(ch3)2 ch2-CH2-N(ch2ch3) 2 CH2-CH2-N^ CHg-CHg-N^] CHg-CHg- o -CH(CH3)OC(-0)C(ch3) 2och3 o «A> 263456 ch3 o A. tBu o 263 45 6 phenyl 408 CH2C(-O)0CH3 409 CH2C(=0)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, p-Ala, Pro, Phe, NMeGly, D-Nle, D-Phg, D-Ile, D-Phe, D-Tyr, Ala.

Claims (13)

-154- VVHATi/l^CLAIM'JS '263456

1. A compound of the formula (I); k^l m / 1 o^\ VNR2 \ PHR\ (CH2)nV R10 (I) or a pharmaceutically acceptable salt form thereof wherein: R1 is H, C2.-C4 alkyl, phenyl, benzyl or phenyl (C1-C4)alkyl; R2 is H or methyl; R10 is H, halogen, Ci-Ce alkyl, phenyl or C1-C4 alkoxy; n is 0-2; n' is 0-1; J is D-Ala, D-Val, D-Ile, D-Leu, D-Nle, D-phenylGly, D-Phe, D-Lys, D-Orn, D-Met, D-Pro, p-Ala, D-Tyr, D-Ser, NMeGly, D-cyclohexylGly, D-cyclohexylmethylGly, D-norvaline, D-2-aminobutyric acid, 0-2-aminopentanoic acid, Gly, N®-p-azidobenzoyl-D-Lys, l^-p-benzoylbenzoyl-D-Lys, K^-tryptophanyl-D-Lys, N^-o-benzylbenzoyl-D-Lys, Ne-p-acetobenzoyl-D-Lys, Ne-dansyl-D-Lys, Ne-t-butoxycarbonylgrlycyl-D-Lys, N^-glycy l-D-Lys, Ne-p-benzoylbenzoylglycyl-D-Lys, Ne-p-phenylbenzoyl-D-Lys, N®-/n-benzoylbenzoyl-D-Lys, or Ne-o-benzoylbenzoyl-D-Lys; -155- 263 45 6 K is CCNMeArg, N^N^diMe-N'-guanidinylOrn, N4*—MeLys or Ne, Na-diMeLys; L is Gly; M is selected from the group consisting of: Asp-(methyicarbonyloxymethyl ester); Asp-(ethylcarbonyloxymethyl ester); Asp- (t-butylcarbonyloxymethyl ester); Asp-(cyclohexylcarbonyloxymethyi eater); 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-(cyclohexylcarbonyloxymethyi 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)-lf3-dioxa-cyclopenten-2-one-4-yl)methyl ester); Asp-((l,3-dioxa-5-phenyl-cyclopenten-2-one-4-y1)methyl ester); and Asp-(1-(2-(2-methoxypropyl)carbonyloxy)ethyl ester),

2. A compound of formula (II): -156- k nr2 26345 CHR1^ (II) or a pharmaceutically acceptable salt form thereof wherein: R1 is H, C1-c4 alkyl, phenyl, benzyl or phenyl(C1-c4) alkyl; R2 is H or methyl; R10 is H, halogen, Ci-Cg-alkyl, phenyl or C1-C4 alkoxy; alternatively, when R10 is para to the carbonyl, R10 and R1 may be taken together to form -CH2-CH2-CH2- thereby to form a six-membered fused ring; n is 0-1; j is D-Ala, D-val, D-Ile, D-Leu, D-Nle, D-phenylGly, D-Phe, D-Lys, D-Orn, D-Met, D-Pro, ^-Ala, D-Tyr, D-Ser, NMeGly, D-cyclohexylGly, D-cyclohexylmethylGly, D-norvaline, D-2-aminobutyric acid, D-2-amlnopentanoic acid, Ne-p-azidobenzoyl-D-Lys, Ne-jp-benzoylbenzoyl-D-Lys, Nr'-tryptophanyl-D-Lys, Nc-o-benzylbenzoyl-D-Lys, Ne-p-acetobenzoy1-D-Lys, Ne-dansy1-D-Lys, NE-t-butoxycarbonylglycyl-D-Lys, N8-glycyl-D-Lys, Nc-p-benzoylbenzoylglycyl-D-Lys, Ne-p-phenylbenzoyl-D-Lys, N£-m-benzoylbenzoyl-D-Lys, or Ne-o-benzoylbenzoyl-D-Lys; K is aNMeArg, N5,Na-diMe-N'-guanidinylOrn, Na-MeLys or Ne,Na-diMeLys; n.Z. patent office 1- OCT 1996 .J received -157- L is Gly; and 263456 M is selected from the group consisting of: Asp-(methyicarbonyloxymethyl ester); Asp-(ethylcarbonyloxymethyl ester); Asp-(t-butylcarbonyloxymethyl ester); Asp-(cyclohexylcarbonyloxymethyi ester); Asp-(1-(methylcarbonyloxy)ethyl ester) ; Asp-(1-(ethylcarbonyloxy)ethyl ester); Asp-(1-(t-butylcarbonyloxy)ethyl ester); Asp-(1-(cyclohexylcarbonyloxy)ethyl ester); Asp-(i-propy loxycarboriyloxymethyl ester); Asp-(cyclohexylcarbonyloxymethyi 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-y 1) methyl ester); and Asp-(1-(2-(2-methoxypropyl)carbonyloxy)ethyl ester).

3. A compound of Claim 2, or a pharmaceutically acceptable salt form thereof, wherein: r1 is H, C1-c4 alkyl, phenyl, benzyl or phenyl(C1-c4)alkyl; R2 is H or methyl; R10 is H, halogen, Ci~C8 alkyl, phenyl or C1-C4 alkoxy; -158- 263456 alternatively, when R10 is para to the carbonyl, R10 and R1 may be taken together to form -CH2-CH2-CH2- thereby to form a six-membered fu9ed ring; n is 1; J is D-Ala, D-Val, D-Ile, D-Leu, D-Nle, D-phenylGly, D-Phe, D-Lys, D-Orn, D-Met, D-Pro, J5-Ala, D-Tyr, D-Ser, NMeGly, D-cyclohexylGly, D-cyclohexylmethylGly, D-norvaline, D-2-aminobutyric acid, Ne-p-azidobenzoyl-D-Lys, Ne-p-benzoylbenzoyl-D-Lys, Ne-tryptophanyl-D-Lys, Ne-o-benzylbenzoyl-D-Lys/ Ng-p-acetobenzoyl-D-Lys, Ne-dansyl— D-Lys, N£-t-butoxycarbonylglycyl-D-Lys, Nc-glycyl-D-Lys, Ne-p-benzoylbenzoylglycyl-D-Lys, Ne-p-phenylbenzoyl-D-Lys, Nc-rtl— benzoylbenzoyl-D-Lys, or Nc-o-benzoylben2oyl-D-Lys; K is CtNMeArg; L is Gly; and M is selected from the group coneisting of: Asp-(methylcarbonyloxymethy ester); Asp-(ethylcarbonyloxymethyl ester); Asp-(t-butylcarbonyloxymethyl ester); Asp-(cyclohexylcarbonyloxymethyi 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-(cyclohexylcarbonyloxymethyi ester); Asp-(t-butyloxycarbonyloxymethyl ester); Asp-(1-(i-propyloxycarbonyloxy)ethyl ester); Asp-(1-(cyclohexyloxycarbonyloxy)ethyl ester); Asp-(1-(t-butyloxycarbonyloxy)ethyl ester); Asp-(dimethylaminoethyl estjer) ; Asp-(diethylaminoethyl ester); -159- Asp-((1,3-dioxa-5 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); and Asp-(1-(2-(2-methoxypropyl)carbonyloxy)ethyl ester).

4. A compound of Claim 2, or a pharmaceutically acceptable salt form thereof, wherein: R1 and R2 are independently selected from H or methyl; R10 is H; alternatively, when R10 is para to the carbonyl, R10 and Rl may be taken together to form -CH2-CH2-CH2- thereby to form a six-membered fused ring; j is selected from: D-Val, D-2-antinobutyric acid, D--Leu, D—Ala, D-Pro, D-Ser, D-Lys, p-Ala, NMeGly, D-Nle, D-pheiiylGly, D-Ile, D-Phe, or D-Tyr; K is CtNMeArg; L is Gly; M is selected from the group consisting of: Asp-(methylcarbonyloxymeth ester); Asp-(ethylcarbonyloxymethyl ester); Asp-(t-butylcarbonyloxymethyl ester); Asp-(cyclohexylcarbonyloxymethyi ester); Asp-(1-(methylcarbonyloxy)ethyl ester); Asp-(1-(ethylcarbonyloxy)ethyl ester); Asp-(1-(t-butylcarbonyloxy)ethyl ester); Asp-(1-(cyclohexylcarbonyloxy)ethyl ester); -160- Asp-(i-propyloxycarbonyloxymethyl ester)? Asp-(cyclohexylcarbonyloxymethyi 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); and Asp-(1-(2-(2-methoxypropyl)carbonyloxy)ethyl ester).

5. A compound of Claim 2 of Formula (IV): or a pharmaceutically acceptable salt form thereof, wherein: R1 and R2 are independently selected from H or methyl; R10 is H, halogen, Ci-Cs alkyl, phenyl or C1-C4 alkoxy; k M \ fl10 (IV) n is 1; r - ' -161- 263456 n' is 1; J is selected from: D-Val, D-2-aminobutyric acid/ D-Leu, D-Ala, D-Pro, D-Serr D-Lys, 0-Ala, NMeGly, D-Nle, D-phenylGly, D-lle, D-Phe, or D-Tyr; K is CtNMeArg; L is Gly; M is selected from the group consisting of: Asp-(methylcarbonyloxymethy ester); Asp-(ethylcarbonyloxymethyl ester); Asp-(t-butylcarbonyloxymethyl ester); Asp-(cyclohexylcarbonyloxymethyi 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-(cyclohexylcarbonyloxymethyi ester); Asp-(t-butyloxycarbonyloxymethyl ester); Asp-(1-(i-propyloxycarbonyloxy)ethyl ester); Asp-(1-(cyclohexyloxycarbonyloxy)ethyl ester); Asp-(l-(t-butyloxycarbonyloxy)ethyl ester); Asp-(dimetbylaminoetbyl ester); Asp-(diethylanunoethyl 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); and Asp—(1-(2-(2-methoxypropyl)carbonyloxy)ethyl ester). r N.Z. PATENT OFFICE 1- OCT 1996 -16 2- 263456

6. A compound of Claim 1 of formula (Ila): (Ila) or a pharmaceutically acceptable salt form thereof, which is selected from the group consisting of: the compound of formula (Ila) wherein R^, R2 and R10 are H; J is D-Val; K is NMeArg; L is Gly; and M is Asp-(methyicarbonyloxymethyl ester); the compound of formula (Ila) wherein R^, R^ and R10 are H; J is D-Val; K is NMeArg; L is Gly; and M is Asp-(ethylcarbonyloxymethyl ester) ; the compound of formula (Ila) wherein R1, R2 and are H; J is o-val; K is NMeArg; L is Gly: and M is Asp-(t-butylcarbonyloxymethyl ester); the compound of formula (Ila) wherein R^, r2 and R1^ are H; J is D-Val; K is NMeArg; L is Gly; and M is Asp- (cyclohexylcarbonyloxymethyi ester); the compound of formula (Ila) wherein R1, R2 and R10 are H; J is D-Val; K is NMeArg; L is Gly; and M is Asp-(1-(methylcarbonyloxy)ethyl ester); the compound of formula (Ila) wherein R1, R2 and R10 are H; J is D-Val; K is NMeArg; L is Gly; and M is Asp-(1-(ethylcarbonyloxy)ethyl ester); 263456 the compound of formula (Ila) wherein R^, R2 and R10 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 (lia) wherein r^, R2 and R10 are H; J is D-Val; K is NMeArg; L is Gly; and M is Asp-(1-(cyclohexylcarbonyloxy)ethyl ester); the compound of formula (Ila) wherein R^, R2 and R10 are H; J is D-Val; K is NMeArg; L is Gly; and M is Asp-(i-propyloxycarbonyloxymethyl ester); the compound of formula (Ila) wherein R1, R2 and R10 are H; J is D-Val; K is NMeArg; L is Gly; and M is Asp-(cyclohexylcarbonyloxymethyi ester); the compound of formula (Ila) wherein R1, R2 and R10 are H; J is D-Val; K is NMeArg; L is Gly; and M is Asp-(t-butyloxycarbonyloxymethyl ester); the compound of formula (Ila) wherein R^-, R2 and R10 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 (Ila) wherein R^-, R2 and R10 are H; J is D-Val; K is NMeArg; L is Gly; and M is Asp-(1-(cyclohexyloxycarbonyloxy)ethyl ester); the compound of formula (Ila) wherein R1, R2 and R10 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 (Ila) wherein R1, R2 and R10 are H; J is D-Val; K is NMeArg; L is Gly; and M is Asp-(dimethylaminoethyl ester); -164- 263456 the compound of formula (Ila) wherein Ri, r2 and R10 are H; J is D-Val; K is NMeArg; L is Gly; and M is Asp-(diethylaminoethyl ester) ; the compound of formula (Ila) wherein R1, R^ and R10 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 (Ila) wherein R^, R^ and R1^ 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 (Ha) wherein R1, R2 and R1() 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 (Ila) wherein R1, R2 and R10 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 (Ila) wherein R1, R2 and R10 are H; J is D-2-aminobutyric acid; K is NMeArg; L is Gly; and M is Asp-(methyicarbonyloxymethyl ester); the compound of formula (Ila) wherein R1, R2 and R10 are H; J is D-2-aminobutyric acid; K is NMeArg; L is Gly; and M is Asp-(ethylcarbonyloxymethyl ester); the compound of formula (Ila) wherein R1, and R10 are H; J is D-2-aminobutyric acid; K is NMeArg; L is Gly; and M is Asp-(t-butylcarbonyloxymethyl ester); the compound of formula (Ila) wherein rI, r2 and Ri0 are H; J is D-2-aminobutyric acid; K is NMeArg; L is Gly; and M is Asp-(cyclohexylcarbonyloxymethyi ester); ' c 263451 the compound of formula (Ila) wherein R1, R2 and R10 are H; J is D-2-aminobutyric acid; K is NMeArg; L is Gly; and M is Asp-(1-(ntethylcarbonyloxy) ethyl ester) / the compound of formula (Ila) wherein rI, R2 and R10 are H; J is D-2-aminobutyric acid; K is NMeArg; L is Gly; and M is Asp-(1-(ethylcarbonyloxy)ethyl ester); the compound of formula (Ila) wherein R1, R2 and R10 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 (Ila) wherein R.1, R2 and R10 are H; J is D-2-aminobutyric acid; K is NMeArg; L is Gly; and M is Asp-(1-(cyclohexylcarbonyloxy)ethyl ester); the compound of formula (Ila) wherein R*-, R2 and R1^ 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 (Ila) wherein R1, R2 and R10 are H; J is o-2-aminobutyric acid; K is NMeArg; L is Gly; and M is Asp-(cyclohexylcarbonyloxymethyi ester); the compound of formula (Ila) wherein R^, R2 and R10 are H; J is D-2-arftinobutyric acid; K is NMeArg; L is Gly; and M is Asp-(t-butyloxycarbonyloxymethyl ester); the compound of formula (Ila) wherein R1, R2 and R10 are H; J is D-2-aminobutyric acid; K is NMeArg; L is Gly; and M is Asp-(l-(i-propyloxycarbonyloxy)ethyl ester); the compound of formula (Ila) wherein R^-, R2 and R10 are H; J is D-2-aminobutyric acid/ K is NMeArg; L is Gly; and M is Asp-(1-(cyclohexyloxycarbonyloxy)ethyl ester); N.Z. PATENT OFRCe 1- 0CT 1996 -166- 263456 the compound of formula (Ila) wherein R1, R2 and R10 are H; J is D-2-aminobutyric acid; K is NMeArg; L is Gly; and M is Asp-(l-(fc-butyloxycarbonyloxy)ethyl ester); the compound of formula (Ila) wherein R^, R2 and R10 are H; J is D-2-aminobutyric acid; K is NMeArg; L is Gly/ and M is Asp-(dimethylaminoethyl ester); the compound of formula (Ila) wherein R1, R2 and R10 are H; J is o-2-aminobutyric acid; K is NMeArg; L is Gly; and M is A3p-(diethyiaminoethyl ester); the compound of formula (Ila) wherein R1, R2 and R10 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 eater); the compound of formula (Ila) wherein R1, R2 and R10 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 (Ila) wherein R1, R2 and R10 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-on<s-4-yl)methyl ester); the compound of formula (Ha) wherein r1, r2 and r10 are h; J is D-2-aminobutyric acid; K is NMeArg; L is Gly; and M is Asp-(l-(2-(2-methoxypropyl)carbonyloxy)ethyl ester).

7. a compound of Formula (I): -167- 263 456 (i) or a pharmaceutically acceptable salt form thereof wherein: r1 is h, c1-c4 alkyl, phenyl, benzyl or phenyl (c3.-c4)alkyl; R2 is H or methyl; r10 is H, halogen, Ci-Cg alkyl, phenyl or C1-C4 alkoxy; n is 0-2; n» is 0-1; J is Ala, Val, lie, Leu, Nle, phenylGly* Phe, Lys, Om, Met, Pro, p-Ala, Tyr, Ser, NMeGly, cyclohexylGly, cyclohexylmethylGly, norvaline, 2-aminobutyric acid, 2-arainopentanoic acid/ Gly, Cys, S-benzyl-Cys, S-methyl-Cys, Asp, Glu, 2-amino-2-methylpropionic acid, His, 1-allo-isoleucine, Asn, Gin, Thr, Trp, or o-methyl-Tyr; K is Arg, N8-Me-N8-guanidinylOrn, p-aminoraethylPhe, p-guanidinylPhe, Lys or N8-MeLys; L ia Gly; M is selected from the group consisting of: nXpatenT'ofrcie 1- OCT 1996 -168- 2634 Asp-(methyicarbonyloxymethyl ester); Asp-(ethylcarbonyloxymethyl ester); Asp-(t-butylcarbonyloxymethyl ester); Asp-(cyclohexylcarbonyloxymethyi 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-(cyclohexylcarbonyloxymethyi ester); Asp-(t-butyloxycarbonyloxymethyl egter); Asp-(1-(i-propyloxycarbonyloxy)ethyl ester); Asp-(1-(cyclohexyloxycarbonyloxy)ethyl ester); Asp-(l-(t-butyloxycarbonyloxy)ethyl ester); Asp-(dimethylaminoethyl ester); Asp-(diethyiaminoethyl ester); Asp-((1,3-dioxa-5-methyl-cyclopenten-2-one-4-yl)methyl ester); Asp-((5-(t-butyl)-l/3-dioxa-cyclopenten-2-one~4-yl)methyl ester); Asp-((1,3-dioxa~5-phenyl-cyclopenten-2-one-4-yl)methyl ester); and Asp-(1-(2-(2-methoxypropyl)carbonyloxy)ethyl ester). NX PATENT Or FACE -169- '263 45 6

8. A compound of Claim 7 of Formula (II): r10 (II) or a pharmaceutically acceptable salt form thereof wherein: r1 is H, C1-C4 alkyl, phenyl, benzyl or phenyl<Ci-C<i)alkyl; R2 is H or methyl; r10 is h, halogen, Ci-Cs alkyl, phenyl or ci-c< alkoxy; alternatively, when R10 is para to the carbonyl, R1(J and R1 may be taken together to form -CH2-CH2-CH2- thereby to form a six-membered fused ring; n is 0—1; J is Ala, Val, lie, Leu, Nle, phenylGly, Phe, Lys, Orn, Met* Pro, |J-Ala, Tyr, Ser, NMeGly, cyclohexylGly, cyclohexylmethylGly, norvaline, 2-aminobutyric acid, 2-aminopentanoic acid, Gly/ Ne-p-azidobenzoyl-Lys, Cys, S-benzyl-Cys, S-methyl—Cys, Asp, Glu, 2-arnino-2-methylpropionic acid, His, 1-allo-isoleucine, Asn, Gin, Thr, Trp, or O-methyl-Tyr; K is Arg, N8-Me-N*-guanidinylOrn, Lys or Nc-MeLya; L is Gly; and M is Selected from the group consisting of: -170- 263 456 Asp-(methyicarbonyloxymethyl ester); Asp-(ethylcarbonyloxymethyl ester); Asp-(t-butylcarbonyloxymethyl ester); Asp-(cyclohexylcarbonyloxymethyi ester); Asp-(1-(methylcarbonyloxy)ethyl ester); Asp-(1-(ethylcarbonyloxy)ethyl ester); Asp-(1-(t-butylcarbonyloxy)ethyl ester); Asp-(1-(cyclohexylcarbonyloxy)ethyl ester); Asp-(i-propyloxycarbonyloxyraethyl ester); Asp-(cyclohexylcarbonyloxymethyi 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-(diethyiaminoethyl 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); and Asp-(1-(2-(2-methoxypropyl)carbonyloxy)ethyl e3ter).

9. A compound of Claim 7 wherein: n is 1; K is Arg; j is Ala, Val, lie. Leu, Pro, Ser, or Lys.

10. A pharmaceutical composition comprising a compound of Claims 1-9 and a pharmaceutically acceptable carrier.

11. A compound of claim 1, claim 2 or claim 3 as specifi set forth herein. B^atent office] 1-OCT1996 Rccavio" 263 456

12. A pharmaceutical composition comprising a compound of any one of claims 1-9 and a pharmaceutically acceptable carrier substantially as herein described.

13. A process for the preparation of a compound of any one of claims 1-9 substantially as herein described with reference to the Examples.