Classifications

• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07K—PEPTIDES
  • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
  • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
  • C07K14/745—Blood coagulation or fibrinolysis factors
  • C07K14/75—Fibrinogen
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P7/00—Drugs for disorders of the blood or the extracellular fluid
  • A61P7/02—Antithrombotic agents; Anticoagulants; Platelet aggregation inhibitors
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K38/00—Medicinal preparations containing peptides

Definitions

• This invention relates to novel cyclic compounds containing carbocyclic ring systems useful as antagonists of the platelet glycoprotein Ilb/IIIa complex, to pharmaceutical compositions containing such cyclic compounds, with or without other therapeutic agents, and to methods of using these compounds, with or without other therapeutic agents, for the inhibition of platelet aggregation, as thrombolytics, and/or for the treatment of other thromboembolic disorders.
• Activation of platelets and the resulting platelet aggregation and secretion of factors by the platelets has been associated with different pathophysiological conditions including cardiovascular and cerebrovascular thromboembolic disorders, for example, the thromboembolic disorders associated with unstable angina, myocardial infarction, transient ischemic attack, stroke, atherosclerosis and diabetes.
• cardiovascular and cerebrovascular 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 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 arterial thrombosis. Platelet activation has been shown to be enhanced during coronary thrombolysis which can 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.
• 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.
• GPIIb/IIIa platelet glycoprotein Ilb/IIIa complex
• GPIIb/IIIa 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 antibody for GPIIb/IIIa indicate the antithrombotic benefit of a GPIIb/IIIa antagonist.
• GPIIb/IIIa-specific antiplatelet agent which inhibits the activation and aggregation of platelets in response to any agonist.
• Such an agent should represent a more efficacious antiplatelet therapy than the currently available agonist-specific platelet inhibitors.
• GPIIb/IIIa does not bind soluble proteins on unstimulated platelets, but GPIIb/IIIa in activated platelets is known to bind four soluble adhesive proteins, namely fibrinogen, von Willebrand factor, fibronectin, and vitronectin.
• fibrinogen The binding of fibrinogen and von Willebrand factor to GPIIb/IIIa causes platelets to aggregate.
• the binding of fibrinogen is mediated in part by the Arg-Gly-Asp (RGD) recognition sequence which is common to the adhesive proteins that bind GPIIb/IIIa.
• This invention provides novel cyclic compounds containing carbocyclic ring systems useful as antagonists of the platelet glycoprotein Ilb/IIIa complex, pharmaceutical compositions containing such cyclic compounds, and methods of using these compounds, alone or in combination with other therapeutic agents, for the inhibition of platelet aggregation, as thrombolytics, and/or for the treatment of thromboembolic disorders.
• This invention also relates to combination products, that is, pharmaceutical compositions containing the novel cyclic compounds of the invention in combination with anti-coagulants such as warfarin or heparin, or anti-platelet agents such as aspirin, piroxicam or ticlopidine, or thrombin inhibitors such as boropeptides, hirudin or argatroban, or thrombolytic agents such as tissue plasminogen activator, anistreplase, urokinase or streptokinase, or combinations thereof, to pharmaceutical kits containing these combination products, and to methods of using these combination products for the inhibition of platelet aggregation, as thrombolytics, and/or for the treatment of thromboembolic disorders.
• anti-coagulants such as warfarin or heparin
• anti-platelet agents such as aspirin, piroxicam or ticlopidine
• thrombin inhibitors such as boropeptides, hirudin or argatroban
• thrombolytic agents
• Figure I shows the platelet 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 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 IC 50 of about 0.0005 mM for Compound A.
• the linear peptide RGDS was much less effective as a thrombolytic, even at substantially higher concentrations .
• 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) , urokinase (UK) and streptokinase (SK) on an already formed platelet-rich clot . The clot was formed by incubating the platelets with agonists for 30 minutes. Both Compounds A and B showed a significant thrombolytic effect as compared to the standard thrombolytics tissue plasminogen activator, urokinase, and streptokinase.
• tPA tissue plasminogen activator
• urokinase UK
• streptokinase streptokinase
• Figure III shows the thrombolytic effect of the cyclic compound cyclo- (D-AbuNMeArg-Gly- Asp-Mamb) (Compound A) and the standard thrombolytics tissue plasminogen activator (tPA) , urokinase (UK) , and streptokinase (SK) , both alone and in combination, on an already formed platelet-rich clot .
• the clot was formed by incubating the platelets with agonists for 30 minutes.
• Compound A showed a significant thrombolytic effect, providing significant clot lysis at 1.0 uM.
• 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 agents administered alone.
• Figure IV shows the thrombolytic effect of the cyclic Ilb/IIIa antagonist compound cyclo- (D-Val- NMeArg-Gly-Asp-Mamb) (Compound B) and the standard thrombolytics tissue plasminogen activator (tPA) , urokinase (UK) and streptokinase (SK) , both alone and in combination, on an already formed platelet-rich clot. The clot was formed by incubating the platelets with agonists for 30 minutes. Compound B showed a significant thrombolytic effect, providing significantly better clot lysis than tissue plasminogen activator, urokinase or streptokinase. Moreover, Compound B in combination with tissue plasminogen activator, urokinase or streptokinase was significantly better than Compound B alone, and significantly better than the additive effects of both agents.
• tPA tissue plasminogen activator
• Figure V shows the thrombolytic effect of the cyclic compound cyclo- (DVal-NMeArg-Gly-Asp- MeMamb) (isomer 1; the compound of Example 68) (Compound C) alone and in combination with the standard thrombolytics tissue plasminogen activator (tPA) , urokinase (UK) and streptokinase (SK) on an already formed platelet-rich clot . The clot was formed by incubating the platelets with agonists for 30 minutes. Compound C alone showed a significant thrombolytic effect. In combination with tissue plasminogen activator, urokinase or streptokinase, a thrombolytic effect was achieved which was greater than the additive effect of the agents when administered alone.
• tissue plasminogen activator urokinase or streptokinase
• Figure VI shows the thrombolytic effect of the cyclic compound cyclo- (D-Val-NMeArg-Gly-Asp- MeMamb) (isomer 2; the compound of Example 68a) (Compound D) alone and in combination with the standard thrombolytics tissue plasminogen activator (tPA) , urokinase (UK) and streptokinase (SK) on an already formed plateletrich clot . The clot was formed by incubating the platelets with agonists for 30 minutes. Compound D alone showed a significant thrombolytic effect. In combination with tissue plasminogen activator, urokinase or streptokinase, a thrombolytic effect was achieved which was greater than the additive effect of the agents when administered alone.
• tissue plasminogen activator urokinase or streptokinase
• Figure VII shows the in vivo thrombolytic and anti-thrombotic effect of the cyclic glycoprotein Ilb/IIIa compound cyclo- (D-Abu-NMeArg-Gly- AspMa b) (Compound A) , alone or in combination with the standard thrombolytic streptokinase (SK) .
• the experiments were carried out using an arterial thrombosis animal model.
• Figure VII shows the results of initial administration as a percentage of clot lysis .
• Figure VIII shows the results of administration of Compound A or saline following streptokinase (SK) or tissue plasminogen activator (t-PA) thrombolysis, with the results reported as time to reocclusion and percentage of reocclusion.
• the saline control showed 100% reocclusion, whereas administration of Compound A resulted in virtually no reocclusion.
• R 31 is a C6-C 14 saturated, partially saturated, or aromatic carbocyclic ring system substituted with 0-4 R 10 or R 10a ;
• R 32 is selected from:
• Z is S or 0
• n" and n' are independently 0-2;
• R 1 and R 22 are independently selected from the following groups:
• heterocyclic ring system containing 1-4 heteroatoms independently selected from N, S, and 0, said heterocyclic ring being substituted with 0-2 R 12 ;
• R 1 and R 21 can alternatively join to form a 3-7 membered carbocyclic ring substituted with 0-2 R 12 ;
• R 1 or R 21 can alternatively be taken together with R 1 or R 21 on an adjacent carbon atom to form a direct bond, thereby to form a double or triple bond between said carbon atoms;
• R22 and R 23 can alternatively join to form a 3-7 membered carbocyclic ring substituted with 0-2 R 12 ;
• R22 or R 23 can alternatively be taken together with R Q r R 23 on an adjacent carbon atom to form a direct bond, thereby to form a double or triple bond between the adjacent carbon atoms;
• R 1 and R 2 where R 21 is H, can alternatively join to form a 5-8 membered carbocyclic ring substituted with 0-2 R 12 ;
• R 11 is selected from one or more of the following:
• C1-C5 alkyl C2-C4 alkenyl, C3-C6 cycloalkyl, C3-C6 cycloalkylmethyl, C2-C6 alkoxyalkyl, C3-C 6 cycloalkoxy, C 1 -C 4 alkyl (alkyl being substituted with 1-5 groups selected independently from:
• heterocyclic ring system containing 1-4 heteroatoms independently selected from N, S, and 0, said heterocyclic ring being substituted with 0-2 R 12 ;
• R 1 2 is selected from one or more of the following:
• R 13 is selected independently from: H, C1-C10 alkyl, C3-C10 cycloalkyl, C4-C12 alkylcycloalkyl, aryl, -(C1-C10 alkyl)aryl, or C3-C10 alkoxyalkyl;
• R 13a is C1-C10 alkyl, C3-C10 cycloalkyl, C4-C12 alkylcycloalkyl, aryl, -(C1 . -C10 alkyl)aryl, or C3-C10 alkoxyalkyl;
• R 13 groups when two R 13 groups are bonded to a single N, said R 13 groups may alternatively be taken together to form - ⁇ CH 2 )2-5- or -(CH 2 )0(CH 2 )-;
• R 14 is OH, H, C1-C4 alkyl, or benzyl
• R 21 and R 23 are independently selected from:
• R 3 is H or Ci-Ce alkyl
• R 4 is H or C1-C3 alkyl
• R5 is selected from: hydrogen
• R 3 and R5 can alternatively be taken together to form
• R 16 is selected from: an amine protecting group; 1-2 amino acids;
• R 6 is H or Ci-Ce alkyl
• R 7 is selected from:
• w.herein eac.h q is independently 0-2 and substitution on the phenyl is at the 3 or 4 position; wherein each q is independently 0-2 and substitution on the cyclohexyl is at the 3 or 4 position;
• X is selected from:
• R6 and R 7 can alternatively be taken together to form
• n 0 or 1 and X is -NH2 or
• M is a D-isomer or L-isomer amino acid of structure
• R 17 is H, C 1 -C3 alkyl
• R8 is selected from:
• -PO(OR 13 )R 13 -S ⁇ 2 NH-heteroaryl (said heteroaryl being 5-10-membered and having 1-4 heteroatoms selected independently from N, S, or 0) , -S ⁇ 2 NH-heteroaryl (said heteroaryl being 5-10-membered and having 1-4 heteroatoms selected independently from N, S, or 0), -SO2NHCOR 13 , -C0NHS0 2 R 13a , -CH 2 CONHS02R 13a , -NHS ⁇ 2 NHCOR 13a , -NHCONHS ⁇ 2R 13a , -SO 2 NHCONHR 13 , -C ⁇ 2R 13b ;
• R 34 and R 3 ⁇ are independently selected from: -OH, -F,
• R 34 and R 3 ⁇ can alternatively be taken together form: a cyclic boron ester where said chain or ring contains from 2 to 20 carbon atoms and, optionally, 1-4 heteroatoms independently selected from N, S, or O; a divalent cyclic boron amide where said chain or ring contains from 2 to 20 carbon atoms and, optionally, 1-4 heteroatoms independently selected from N, S, or 0; a cyclic boron amide-ester where said chain or ring contains from 2 to 20 carbon atoms and, optionally, 1-4 heteroatoms independently selected from N, S, or 0;
• R 13b is selected from:
• R 36 is selected independently from: H, Ci-C ⁇ alkyl, C3-C 1 0 cycloalkyl, phenyl, or benzyl;
• R 37 is selected from: (a) H; (b) Ci-C ⁇ alkyl or C3-C8 cycloalkyl, said alkyl or cycloalkyl being substituted with 1-2 groups independently selected from:
• aryl substituted with 0-2 groups independently selected from: halogen, phenyl,
• R 38 is selected from: (a) Ci-C ⁇ alkyl or C3-C8 cycloalkyl, said alkyl or cycloalkyl being substituted with 1-2 groups independently selected from: (i) C1-C4 alkyl; (ii) C3-C8 cycloalkyl;
• aryl substituted with 0-2 groups independently selected from: halogen, phenyl, C1-C6 alkyl, C1-C6 alkoxy, NO2, -S (C1-C5 alkyl), -SO(C ⁇ -Cs alkyl), -S0 2 (C ⁇ -C 5 alkyl),
• Ci-C ⁇ alkyl or C3-C8 cycloalkyl said alkyl or cycloalkyl being substituted with 1-2 groups independently selected from: (i) Ci-C ⁇ alkyl;
• aryl substituted with 0-2 groups independently selected from: halogen, phenyl, C1-C6 alkyl, C1-C6 alkoxy, N0 2 , -S (C1-C5 alkyl), -SO(C ⁇ -C 5 alkyl), -S0 2 (C1-C5 alkyl),
• >40 is selected from: H, C1-C5 alkyl, or benzyl;
• R 8 is -B(R 34 ) (R 35 ) or -C ⁇ 2R 13b .
• the present invention includes the use of the above described compounds (wherein the above conditions (1)- (5) are not required) in combination with one or more additional therapeutic agents for the inhibition of platelet aggregation, as thrombolytics, and/or for the treatment of thromboembolic disorders, wherein the additional therapeutic agent is selected from: anti-coagulants such as warfarin or heparin, or anti ⁇ platelet agents such as aspirin, piroxicam or ticlopidine; thrombin inhibitors such as boropeptides, hirudin or argatroban; or thrombolytic agents such as tissue plasminogen activator, anistreplase, urokinase or streptokinase.
• anti-coagulants such as warfarin or heparin, or anti ⁇ platelet agents
• thrombin inhibitors such as boropeptides, hirudin or argatroban
• thrombolytic agents such as tissue plasminogen activator, anistreplase, urokina
• R 31 is bonded to (C (R 23 ) R 22 ) n " and (C (R 21 ) R 1 ) n ' at 2 different atoms on said carbocyclic ring.
• R 31 is a C6 ⁇ Ci 4 saturated, partially saturated, or aromatic carbocyclic ring system substituted with 0-4 R 10 or R 10a ;
• Z is S or 0
• n" and n' are independently 0-2;
• R 1 and R 22 are independently selected from the following groups:
• heterocyclic ring system containing 1-4 heteroatoms independently selected from N, S, or 0, said heterocyclic ring being substituted with 0-2 R 12 ;
• R and R 21 can alternatively join to form a 5-7 membered carbocyclic ring substituted with 0-2 R 12 ; when n' is 2, R 1 or R 21 can alternatively be taken together with R 1 or R 21 on an adjacent carbon atom to form a direct bond, thereby to form a double or triple bond between said carbon atoms;
• R 22 and R 23 can alternatively join to form a 3-7 membered carbocyclic ring substituted with 0-2 R 12 ;
• R 22 or R 23 can alternatively be taken together with R 22 or R 23 on an adjacent carbon atom to form a direct bond, thereby to form a double or triple bond between said carbon atoms;
• R 1 and R 2 where R 21 is H, can alternatively join to form a 5-8 membered carbocyclic ring substituted with 0-2 R 12 ;
• R 1 is selected from one or more of the following:
• aryl substituted with 0-2 R 12 a 5-10-membered heterocyclic ring system containing 1-4 heteroatoms independently selected from N, S, or 0, said heterocyclic ring being substituted with 0-2 R 12 ;
• ,12 is selected from one or more of the following:
• R 13 is selected independently from: H, C1-C10 alkyl, c 3 ⁇ c 10 cycloalkyl, C4-C12 alkylcycloalkyl, aryl, - (C1-C10 alkyl) aryl, or C3-C10 alkoxyalkyl;
• R 13a is C1-C10 alkyl, C3-C10 cycloalkyl, C4-C12 alkylcycloalkyl, aryl, -(C1-C10 alkyl)aryl, or C3-C10 alkoxyalkyl;
• R 13 groups when two R 13 groups are bonded to a single N, said R 13 groups may alternatively be taken together to form -(CH 2 )2- 5 - or -(CH 2 )0(CH 2 )-;
• R 14 is OH, H, C1-C4 alkyl, or benzyl;
• R 21 and R 23 are independently selected from:
• R 2 is H or C 1 -C8 alkyl
• R 1( ⁇ and R 10a are selected independently from one or more of the following:
• R 4 is H or C1-C3 alkyl
• R 16 is selected from: an amine protecting group; 1-2 amino acids; 1-2 amino acids subtituted with an amine protecting group;
• R 6 is H or Ci-C ⁇ alkyl
• R 7 is selected from:
• each q is independently 0-2 and substitution on the phenyl is at the 3 or 4 position;
• R" and R 7 can alternatively be taken together to form
• q 1 is 0-2 ;
• R 17 is H, C1-C3 alkyl ;
• R ⁇ is selected from:
• R 34 and R 3 ⁇ are independently selected from: -OH, -F,
• R 34 and R 3 ⁇ can alternatively be taken together form: a cyclic boron ester where said chain or ring contains from 2 to 20 carbon atoms and, optionally, 1-4 heteroatoms independently selected from N, S, or 0; a divalent cyclic boron amide where said chain or ring contains from 2 to 20 carbon atoms and, optionally, 1-4 heteroatoms independently selected from N, S, or 0; a cyclic boron amide-ester where said chain or ring contains from 2 to 20 carbon atoms and, optionally, 1-4 heteroatoms independently selected from N, S, or 0;
• R l3b j_ s selected from:
• aryl optionally substituted with 1-2 substituents independently selected from halogen, phenyl, C1-C5 alkyl,
• R 36 is selected independently from: H, Ci-Cs alkyl, C3-C10 cycloalkyl, phenyl, or benzyl;
• R 37 is selected from:
• Ci-C ⁇ alkyl or C3-C8 cycloalkyl said alkyl or cycloalkyl being substituted with 1-2 groups independently selected from: (i) C1-C4 alkyl;
• aryl substituted with 0-2 groups independently selected from: halogen, phenyl, Ci-Cg alkyl,
• R 38 is selected from:
• R 39 is selected from:
• Ci-C ⁇ alkyl or C3-C8 cycloalkyl said alkyl or cycloalkyl being substituted with 1-2 groups independently selected from: (i) Ci-C ⁇ alkyl;
• aryl substituted with 0-2 groups independently selected from: halogen, phenyl, Ci-C ⁇ alkyl, C1-C6 alkoxy, NO2, -S (C1-C5 alkyl), -S0(C ⁇ -Cs alkyl), -SO2 (C1-C5 alkyl),
• R 40 is selected from: H, C1-C5 alkyl, or benzyl.
• R 31 is selected from the group consisting of (a) a 6 membered saturated, partially saturated or aromatic carbocyclic ring substituted with 0-3 R 10 or R 10a ;
• R 31 is selected from the group consisting of:
• any of the bonds forming the carbocyclic ring may be a single or double bond. and wherein said carbocyclic ring is substituted independently with 0-4 R iO ;
• any of the bonds forming the carbocyclic ring may be a single or double bond
• any of the bonds forming the carbocyclic ring may be a single or double bond
• This invention includes compounds of formula (I) wherein:
• R 31 is selected from (the dashed bond may be a single or double bond) :
• R 31 may be substituted independently with 0-3 R 10 or R 10a ;
• n" is 0 or 1; n' is 0-2.
• the present invention includes compounds of formula (I), or a pharmaceutically acceptable salt or prodrug form thereof wherein:
• R 31 is selected from:
• R 31 may be substituted independently with 0-3 R 10 or R 10a ; n " is 0 or 1 ; n 1 is 0-2 ;
• R and R 22 are independently selected from H, C1-C4 alkyl, phenyl, benzyl, phenyl- (C2-C4)alkyl, C 1 -C 4 alkoxy;
• R 21 and R 23 are independently H or C 1 -C 4 alkyl
• R 2 is H or Ci-C ⁇ alkyl
• R 13 is selected independently from: H, C1-C10 alkyl, C3-C10 cycloalkyl, C4-C12 alkylcycloalkyl, aryl, -(Ci-Cio alkyl) aryl, or C3-C10 alkoxyalkyl;
• R 13a is C1-C10 alkyl, C3-C10 cycloalkyl, C4-C12 alkylcycloalkyl, aryl, - (C1-C3 . 0 alkyl)aryl, or
• R 13 groups when two R 13 groups are bonded to a single N, said R 13 groups may alternatively be taken together to form
• R 14 is OH, H, C1-C4 alkyl, or benzyl
• R 1 *-- 1 and R 10a are selected independently from: H, Ci-C ⁇ alkyl, phenyl, halogen, or C 1 -C 4 alkoxy;
• R 3 is H or CH3;
• R 4 is H or C1-C3 alkyl;
• R 16 is selected from: an amine protecting group; 1-2 amino acids; or
• R 6 is H or Ci-C ⁇ alkyl
• R6 and R 7 can alternatively be taken together to form
• M is a D-isomer or L-isomer amino acid of structure
• q' is 0-2;
• R 17 is H, C1-C3 alkyl;
• R ⁇ is selected from:
• R 13b is selected from:
• R 36 is selected independently from: H, Ci-C ⁇ alkyl, C3-C10 cycloalkyl, phenyl, or benzyl;
• R 37 is selected from: (a) H;
• Ci-C ⁇ alkyl or C3-C8 cycloalkyl said alkyl or cycloalkyl being substituted with 1-2 groups independently selected from:
• aryl substituted with 0-2 groups independently selected from: halogen, phenyl, Ci-C ⁇ alkyl, Ci-C ⁇ alkoxy, NO2, -S (C1-C5 alkyl), -SO(C ⁇ -C 5 alkyl), -SO2 (C1-C5 alkyl),
• R 38 is selected from: (a) CI-C ⁇ alkyl or C3-C8 cycloalkyl, said alkyl or cycloalkyl being substituted with 1-2 groups independently selected from:
• aryl substituted with 0-2 groups independently selected from: halogen, phenyl, C1-C6 alkyl, Ci-C ⁇ alkoxy, NO2, -S (C1-C5 alkyl), -SO(C ⁇ -Cs alkyl), -SO2 (C1-C5 alkyl),
• Ci-C ⁇ alkyl or C3-C8 cycloalkyl said alkyl or cycloalkyl being substituted with 1-2 groups independently selected from: (i) Ci-C ⁇ alkyl;
• aryl substituted with 0-2 groups independently selected from: halogen, phenyl, C1-C6 alkyl, Ci-C ⁇ alkoxy, NO2, -S (C1-C5 alkyl), -SO(C ⁇ -C 5 alkyl), -SO2 (C1-C5 alkyl),
• R 40 is selected from: H, C 1 -C5 alkyl, or benzyl
• Preferred compounds of the invention are 1,3- disubstituted phenyl compounds of the formula (II) :
• the phenyl ring in formula (II) may be further substituted with 0-3 R 10 ;
• R 10 is selected independently from: H, Ci-C ⁇ alkyl, phenyl, halogen, or C 1 -C 4 alkoxy;
• R is H, C1-C4 alkyl, phenyl, benzyl, or phenyl- (C ⁇ C4)alkyl;
• R 2 is H or methyl;
• R 13 is selected independently from: H, C1-C10 alkyl, C3-C10 cycloalkyl, C4-C12 alkylcycloalkyl, aryl, -(C1-C10 alkyl)aryl, or C3-C10 alkoxyalkyl;
• R 13a is Ci-Cio alkyl, C3-C10 cycloalkyl, C4-C12 alkylcycloalkyl, aryl, -(C ⁇ -C ⁇ o alkyl) aryl, or C3-C10 alkoxyalkyl;
• R 13 groups when two R 13 groups are bonded to a single N, said R 13 groups may alternatively be taken together to form ⁇ (CH 2 )2-5- or -(CH 2 )0(CH 2 )-;
• R 14 is OH, H, C1-C4 alkyl, or benzyl
• R 3 is H or CH3
• R 4 is H or C1-C3 alkyl
• R 5 is H, C1-C8 alkyl, C3-C6 cycloalkyl, C3-C6 cycloalkylmethyl, C1-C6 cycloalkylethyl, phenyl, phenylmethyl, CH2OH, CH2SH, CH2OCH3,
• R 16 is selected from: an amine protecting group
• R b is H or C ⁇ -C8 alkyl;
• R 7 is:
• M is a D-isomer or L-isomer amino acid of structure
• R 17 is H, C 1 -C 3 alkyl
• R* is selected from:
• R 13b is selected from:
• aryl optionally substituted with 1-2 substituents independently selected from halogen, phenyl, C1-C5 alkyl,
• R 36 is selected independently from: H, Ci-C ⁇ alkyl, C3-C10 cycloalkyl, phenyl, or benzyl;
• R 37 is selected from: (a) H;
• aryl substituted with 0-2 groups independently selected from: halogen, phenyl, C1-C6 alkyl, Ci-C ⁇ alkoxy, NO2, -S (C1-C5 alkyl), -SO(C ⁇ -C 5 alkyl), -S02(C ⁇ -C 5 alkyl),
• R 38 is selected from: (a) Ci-C ⁇ alkyl or C3-C8 cycloalkyl, said alkyl or cycloalkyl being substituted with 1-2 groups independently selected from: (i) C1-C4 alkyl; (ii) C3-C8 cycloalkyl;
• aryl substituted with 0-2 groups independently selected from: halogen, phenyl, Ci-Ce alkyl, C ⁇ C ⁇ alkoxy, NO2, -S (C1-C5 alkyl), -SO(C ⁇ -C 5 alkyl), -SO2 (C1-C5 alkyl),
• Ci-C ⁇ alkyl or C3-C8 cycloalkyl said alkyl or cycloalkyl being substituted with 1-2 groups independently selected from: (i) Ci-C ⁇ alkyl;
• aryl substituted with 0-2 groups independently selected from: halogen, phenyl, C1-C6 alkyl, C1-C6 alkoxy, NO2, -S(C ⁇ -Cs alkyl), -SO(C ⁇ -Cs alkyl), -SO2 (C1-C5 alkyl),
• R 40 is selected from: H, C 1 -C 5 alkyl, or benzyl.
• Preferred compounds of the present invention are compounds of formula (II) above, wherein:
• the phenyl ring in formula (II) may be further substituted with 0-2 R 10 or R 10a ;
• R 10 or R 10a are selected independently from: H, Ci-C ⁇ alkyl, phenyl, halogen, or C 1 -C 4 alkoxy;
• R 13 is selected independently from: H, C1-C10 alkyl, C3-C10 cycloalkyl, C4-C12 alkylcycloalkyl, aryl, -(C1-C10 alkyl) aryl, or C3-C10 alkoxyalkyl;
• R 13a i s C1-C10 alkyl, C3-C10 cycloalkyl, C4-C12 alkylcycloalkyl, aryl, -(C ⁇ -C ⁇ o alkyl)aryl, or C3-C10 alkoxyalkyl;
• R 13 groups when two R 13 groups are bonded to a single N, said R 13 groups may alternatively be taken together to form -(CH 2 )2-5 ⁇ or -(CH 2 )0(CH 2 )-;
• R 14 is OH, H, C1-C4 alkyl, or benzyl
• R 3 is CH3 and R 5 is H;
• R 3 and R ⁇ can alternatively be taken together to form -CH2CH2CH2-;
• R 16 is selected from: an amine protecting group; 1-2 amino acids;
• M is a D-isomer or L-isomer amino acid of structure
• R 4 is H or CH3
• R 17 is H; -C02H; -S03H ;
• R 36 is C1-C4 linear alkyl or H
• R 37 is selected from:
• R 38 is selected from: (a) Ci-C ⁇ alkyl or C3-C8 cycloalkyl, said alkyl or cycloalkyl being substituted with 1-2 groups independently selected from: (i) C 1 -C 4 alkyl; (ii) C3-C8 cycloalkyl; (iii) C 1 -C5 alkoxy;
• R 39 is C1-C 4 alkyl, benzyl, or phenyl.
• Preferred compounds of the present invention are compounds of formula (II), or a pharmaceutically acceptable salt thereof, wherein:
• R 1 and R 2 are independently selected from H, methyl
• J is selected from D-Val, D-2-aminobutyric acid, D-Leu, D-Ala, Gly, D-Pro, D-Ser, D-Lys, ⁇ -Ala, Pro, Phe,
• NMeGly D-Nle, D-Phg, D-Ile, D-Phe, D-Tyr, Ala, N ⁇ -p-azidobenzoyl-D-Lys, N ⁇ -p-benzoylbenzoyl-D-Lys, N ⁇ -tryptophanyl-D-Lys, N ⁇ -o-benzylbenzoyl-D-Lys, N ⁇ -p-acetylbenzoy1-D-Lys, N ⁇ -dansy1-D-Lys, N ⁇ -glycyl-D-Lys, N ⁇ -glycy1-p-benzoylbenzoy1-D-Lys, N ⁇ -p-phenylbenzoyl-D-Lys, N ⁇ -m-benzoylbenzoyl-D- Lys, N ⁇ -o-benzoyl
• K is selected from NMeArg, Arg;
• L is selected from Gly, ⁇ -Ala, Ala;
• M is selected from Asp; OMeAsp; ⁇ MeAsp; NMeAsp; D-Asp;
• Preferred compounds of the present invention are compounds of formula (II) , or a pharmaceutically acceptable salt thereof, wherein:
• R 1 and R 2 are independently selected from H, methyl
• J is selected from: D-Val, D-2-aminobutyric acid, D-Leu, D-Ala, Gly, D-Pro, D-Ser, D-Lys, ⁇ -Ala, Pro, Phe,
• NMeGly D-Nle, D-Phg, D-Ile, D-Phe, D-Tyr, Ala;
• K is selected from NMeArg
• L is Gly
• M is selected from Asp; OMeAsp; ⁇ MeAsp; NMeAsp; D-Asp;
• Specifically preferred compounds of the present invention are the following compounds and pharmaceutically acceptable salts thereof:
• GPIIb/IIIa glycoprotein Ilb/IIIa
• the compounds of the present invention inhibit the activation and aggregation of platelets induced by all known endogenous platelet agonists.
• the present invention also provides methods for the treatment (including prevention) of conditions involving platelet activation and aggregation, such as arterial or venous cardiovascular or cerebrovascular thromboembolic disorders, including, for example, thromboembolic disorders associated with unstable angina, first or recurrent myocardial infarction, ischemic sudden death, transient ischemic attack, stroke, atherosclerosis, deep vein thrombosis, pulmonary embolism, or diabetes, by administering to a host in need of such treatment a pharmaceutically effective amount of the compounds described above.
• thromboembolic disorders associated with unstable angina, first or recurrent myocardial infarction, ischemic sudden death, transient ischemic attack, stroke, atherosclerosis, deep vein thrombosis, pulmonary embolism, or diabetes
• the compounds of the present invention are useful for inhibiting the binding of fibrinogen to blood platelets, inhibiting aggregation of blood platelets, treating thrombus formation or embolus formation, or preventing thrombus or embolus formation in a mammal.
• the compounds of the invention may be used as a medicament for blocking fibrinogen from acting at its receptor site in a mammal.
• the compounds of the present invention can also be combined or co-administered with suitable anti-coagulant or coagulation inhibitory agents, such as heparin or warfarin, or anti-platelet or platelet inhibitory agents, such as aspirin, piroxicam or ticlopidine. Further, the compounds of this invention may be combined or co-administered with thrombin inhibitors such as boropeptides, hirudin or argatroban. The compounds of the present invention may also be combined or co-administered with thrombolytic or fibrinolytic agents, such as plasminogen activators, anistreplase, urokinase, or streptokinase.
• suitable anti-coagulant or coagulation inhibitory agents such as heparin or warfarin
• anti-platelet or platelet inhibitory agents such as aspirin, piroxicam or ticlopidine.
• thrombin inhibitors such as boropeptides, hirudin or argatroban
• 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.
• platelet granular secretions of various important biomolecules from the ⁇ -granule (PAI- 1) or the dense granule (serotonin) are not affected by the GPIIb/IIIa antagonist.
• These molecules secreted from platelets might play an important role in arterial vasospasm (serotonin) and in reducing the efficiency of the natural lytics (PAI-1) .
• PAI-1 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 heterogen
• the GPIIb/IIIa antagonists of the present invention with high affinity for the platelet GPIIb/IIIa receptor are expected to be very effective not only in preventing thrombosis formation, but also in accelerating lysis of platelet rich thrombi, thereby providing a greater utility of such antiplatelet agents in the acute and chronic thromboembolic disorders.
• Such 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) .
• anti-coagulant agents denotes agents that inhibit blood coagulation.
• agents include warfarin, heparin, or low molecular weight heparin (LM H) , including pharmaceutically acceptable salts or prodrugs thereof.
• the preferable anti-coagulant agents are warfarin or heparin or LMWH.
• the warfarin employed herein may be, for example, crystalline warfarin or amorphous sodium warfarin.
• the heparin employed herein may be, for example, the sodium or sulfate salts thereof.
• anti-platelet agents denotes agents that inhibit platelet function such as by inhibiting the aggregation, adhesion or granular secretion of platelets.
• agents include the various known non-steroidal anti-inflammatory drugs (NSAIDS) such as aspirin, ibuprofen, naproxen, sulindac, indomethacin, mefenamate, droxicam, diclofenac, sulfinpyrazone, and piroxicam, including pharmaceutically acceptable salts or prodrugs thereof.
• NSAIDS non-steroidal anti-inflammatory drugs
• NSAIDS Aspirin (acetylsalicyclic acid or ASA)
• piroxicam which exerts its anti-platelet effect when dosed once daily
• Piroxicam is commercially available from Pfizer Inc. (New York, NY), as FELDANETM.
• Other suitable anti-platelet agents include ticlopidine, including pharmaceutically acceptable salts or prodrugs thereof. Ticlopidine is also a preferred compound since it 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 pharmaceutically acceptable salts or prodrugs thereof
• thrombin inhibitors denotes inhibitors of the serine protease thrombin.
• various thrombinmediated processes such as thrombin-mediated platelet activation (that is, for example, the aggregation of platelets, and/or the granular secretion of plasminogen activator inhibitor-1 and/or serotonin) and/or fibrin formation are disrupted.
• Such inhibitors include boropeptides, hirudin and argatroban, including pharmaceutically acceptable salts and prodrugs thereof.
• the thrombin inhibitors are boropeptides.
• boropeptides N-acetyl and peptide derivatives of boronic acid, such as C-terminal ⁇ -aminoboronic acid derivatives of lysine, ornithine, arginine, homoarginine and corresponding isothiouronium analogs thereof.
• hirudin 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.
• 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.
• thrombolytics or fibrinolytic agents
• fibrinolytics or fibrinolytics agents that lyse blood clots (thrombi) .
• agents include tissue plasminogen activator, anistreplase, urokinase or streptokinase, including pharmaceutically acceptable salts or prodrugs thereof.
• Tissue plasminogen activator (tPA) is commercially available from Genentech Inc., South San Francisco, California.
• anistreplase refers to anisoylated plasminogen streptokinase activator complex, as described, for example, in European Patent Application No.
• urokinase is intended to denote both dual and single chain urokinase, the latter also being referred to herein as prourokinase .
• Combination products where the cyclic compounds of the invention are combined or co-administered with suitable anti-coagulant agents, antiplatelet agents, thrombin inhibitors, and/or thrombolytic agents, may afford an efficacy advantage over the compounds and agents alone, and may do so while permitting the use of lower doses of each.
• a lower dosage minimizes the potential of side effects, thereby providing an increased margin of safety.
• ASA acetylsalicylic acid
• ASA platelet function
• ticlopidine agents that inhibit platelet function
• ASA and ticlopidine have been shown to be effective in the prevention of stroke in people with cerebrovascular disease .
• Pooled data from nine randomized trials have provided overwhelming evidence of the efficacy of ASA alone in reducing the risk of completed stroke in people with transient ischemic attacks (TIAs) .
• TIAs transient ischemic attacks
• ticlopidine alone has also been demonstrated to have efficacy in treating TIAs.
• thrombin inhibitors such as boropeptides
• studies have demonstrated that such compounds provide excellent candidates for the control of thrombinmediated processes.
• Studies with hirudin, another thrombin inhibitor, have shown this agent to be an effective compound in the treatment of venous and arterial thrombosis.
• Current therapy in the treatment of patients with acute myocardial infarction includes thrombolytics such as plasminogen activators such as tPA, streptokinase, or urokinase.
• tPA plasminogen activators
• streptokinase streptokinase
• urokinase 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.
• an anti-coagulant agent such as warfarin or heparin
• an antiplatelet agent such as aspirin, piroxicam or ticlopidine
• a thrombin inhibitor such as a boropeptide, hirudin or argatroban
• a thrombolytic agent such as tissue plasminogen activator, anistreplase, urokinase or streptokinase, or combinations thereof, in combination with a novel
• 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 invention, or combinations thereof, are directed to meeting these, as well as other, needs.
• GPIIb/IIIa is known to be overexpressed in metastatic tumor cells.
• the compounds or combination products of the present invention may also be useful for the treatment, including prevention, of metastatic cancer.
• the D and L-isomers of a particular amino acid are designated herein using the conventional 3- letter abbreviation of the amino acid, as indicated by the following examples: D-Leu, D-Leu, L-Leu, or L-Leu.
• any variable for example, R 1 through R 8 , m, n, p, X, Y, etc.
• its definition on each occurrence is independent of its definition at every other occurrence.
• R 11 for example, if a group is shown to be substituted with 0-2 R 11 , then said group may optionally be substituted with up to two R 11 and R 11 at each occurrence is selected independently from the defined list of possible R 11 .
• the group -N(R 13 )2 each of the two R 13 substituents on N is independently selected from the defined list of possible R 13 .
• stable compound or “stable structure” is meant herein a compound that is sufficiently robust to survive isolation to a useful degree of purity from a reaction mixture, and formulation into an efficacious therapeutic agent .
• substituted 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.
• 2 hydrogens on the atom are replaced.
• alkyl is intended to include both branched and straight-chain saturated aliphatic hydrocarbon groups having the specified number of carbon atoms;
• alkoxy represents an alkyl group of indicated number of carbon atoms attached through an oxygen bridge;
• cycloalkyl is intended to include saturated ring groups, including mono-,bi- or poly-cyclic ring systems, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl and adamantyl;
• biycloalkyl is intended to include saturated bicyclic ring groups such as [3.3.
• Alkenyl is intended to include hydrocarbon chains of either a straight or branched configuration and one or more unsaturated carbon-carbon bonds which may occur in any stable point along the chain, such as ethenyl, propenyl and the like; and "alkynyl” is intended to include hydrocarbon chains of either a straight or branched configuration and one or more triple carbon-carbon bonds which may occur in any stable point along the chain, such as ethynyl, propynyl and the like.
• boronic acid means a group of the formula -B(R 34 ) (R ⁇ ) , wherein R 34 and R 35 are independently selected from: -OH; -F; -NR l3 R 14 ; or Ci-C ⁇ -alkoxy; or R 34 and R 3 ⁇ can alternatively be taken together to form: a cyclic boron ester where said chain or ring contains from 2 to 20 carbon atoms and, optionally, 1-4 heteroatoms independently selected from N, S, or O; a divalent cyclic boron amide where said chain or ring contains from 2 to 20 carbon atoms and, optionally, 1-4 heteroatoms independently selected from N, S, or 0; a cyclic boron amide-ester where said chain or ring contains from 2 to 20 carbon atoms and, optionally, 1-4 heteroatoms independently selected from N, S, or O.
• Boron esters include boronic acid protecting groups, including moieties derived from diols, for example pinanediol and pinacol to form pinanediol boronic acid ester and the pinacol boronic acid, respectively.
• diols useful for deriving boronic acid esters are perfluoropinacol, ethylene glycol, diethylene glycol, 1,2-ethanediol, 1, 3-propanediol, 1, 2-propanediol, 1, 2-butanediol, 1, -butanediol, 2,3-butanediol, 2,3-hexanediol, 1, 2-hexanediol, catechol, 1, 2-diisopropylethanediol, 5, 6-decanediol, 1,2-dicyclohexylethanediol.
• Halo or "halogen” as used herein refers to fluoro, chloro, bromo and iodo; and "counterion” is used to represent a small, negatively charged species such as chloride, bromide, hydroxide, acetate, sulfate and the like.
• aryl or “aromatic residue” is intended to mean phenyl or naphthyl .
• carbocycle or “carbocyclic residue” is intended to mean any stable 3- to 7- membered monocyclic or bicyclic or 7- to 14-membered bicyclic or tricyclic or an up to 26-membered polycyclic carbon ring, any of which may be saturated, partially unsaturated, or aromatic.
• carbocyles include, but are not limited to, cyclopropyl, cyclopentyl, cyclohexyl, phenyl, biphenyl, naphthyl, indanyl, adamantyl, or tetrahydronaphthyl (tetralin) .
• heterocycle or “heterocyclic ring system” is intended to mean a stable 5- to 7- membered monocyclic or bicyclic or 7- to 10- membered bicyclic heterocyclic ring which may be saturated, partially unsaturated, or aromatic, and which consists of carbon atoms and from 1 to 4 heteroatoms selected independently from the group consisting of N, O and S and wherein the nitrogen and sulfur heteroatoms may optionally be oxidized, and the nitrogen may optionally be quaternized, and including any bicyclic group in which any of the above-defined heterocyclic rings is fused to a benzene ring.
• the heterocyclic ring may be attached to its pendant group at any heteroatom or carbon atom which results in a stable structure.
• the heterocyclic rings described herein may be substituted on carbon or on a nitrogen atom if the resulting compound is stable .
• Examples of such heterocycles include, but are not limited to, benzopyranyl, thiadiazine, tetrazolyl, benzofuranyl, benzothiophenyl, indolene, quinoline, isoquinolinyl or benzimidazolyl, piperidinyl, 4-piperidone, 2-pyrrolidone, tetrahydrofuran, tetrahydroquinoline, tetrahydroisoquinoline, decahydroquinoline, octahydroisoquinoline, azocine, triazine (including 1,2,3-, 1,2,4-, and 1, 3, 5-triazine) , 6H-1,2,5- thiadiazine, 2H, 6H-1
• 4aH-carbazole carbazole, ⁇ -carboline, phenanthridine, acridine, perimidine, phenanthroline, phenazine, phenarsazine, phenothiazine, furazan, phenoxazine, isochroman, chroman, pyrrolidine, pyrroline, imidazolidine, imidazoline, pyrazolidine, pyrazoline, piperazine, indoline, isoindoline, quinuclidine, or morpholine . Also included are fused ring and spiro compounds containing, for example, the above heterocycles .
• any group that, when administered to a mammalian subject, cleaves to form a free hydroxyi, amino or sulfhydryl means any group bonded to an 0, N, or S atom, respectively, which is cleaved from the O, N, or S atom when the compound is administered to a mammalian subject to provide a compound having a remaining free hydroxyi, amino, or sulfhydryl group, respectively.
• Examples of groups that, when administered to a mammalian subject, are cleaved to form a free hydroxyi, amino or sulfhydryl include but are not limited to, C ⁇ -C6 alkyl substituted with 0-3 R 11 , C3-C6 alkoxyalkyl substituted with 0-3 R 11 , C 1 -C6 alkylcarbonyl substituted with 0-3 R 11 , C ⁇ -C6 alkoxycarbonyl substituted with 0-3 R 11 , Ci-C ⁇ alkylaminocarbonyl substituted with 0-3 R 11 , benzoyl substituted with 0-3 R 12 , phenoxycarbonyl substituted with 0-3 R 12 , phenylaminocarbonyl substituted with 0-3 R 12 .
• Examples of groups that, when administered to a mammalian subject, are cleaved to form a free hydroxyi, amino or sulfhydryl include hydroxy, amine or sulfhydr
• amine protecting group means any group known in the art of organic synthesis for the protection of amine groups. Such amine protecting groups include those listed in Greene, “Protective Groups in Organic Synthesis” John Wiley & Sons, New York (1981) and “The Peptides: Analysis, Sythesis, Biology, Vol. 3, Academic Press, New York (1981) , the disclosure of which is hereby incorporated by reference. Any amine protecting group known in the art can be used. Examples of amine protecting groups include, but.
• acyl types such as formyl, trifluoroacetyl, phthalyl, and p-toluenesulfonyl
• aromatic carbamate types such as benzyloxycarbonyl (Cbz) and substituted benzyloxycarbonyls, 1- (p-biphenyl) -1- methylethoxycarbonyl, and 9-fluorenylmethyloxycarbonyl (Fmoc)
• 3) aliphatic carbamate types such as tert- butyloxycarbonyl (Boc) , ethoxycarbonyl, diisopropylmethoxycarbonyl, and allyloxycarbonyl
• cyclic alkyl carbamate types such as cyclopentyloxycarbonyl and adamantyloxycarbonyl
• alkyl types such as triphenylmethyl and benzyl
• trialkylsilane such as trimethylsi
• amine protecting group are acyl groups such as azidobenzoyl, p-benzoylbenzoyl, o-benzylbenzoyl, p-acetylbenzoyl, dansyl, glycyl-p-benzoylbenzoyl, phenylbenzoyl, m-benzoylbenzoyl, benzoylbenzoyl.
• acyl groups such as azidobenzoyl, p-benzoylbenzoyl, o-benzylbenzoyl, p-acetylbenzoyl, dansyl, glycyl-p-benzoylbenzoyl, phenylbenzoyl, m-benzoylbenzoyl, benzoylbenzoyl.
• 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) .
• pharmaceutically acceptable salts include, but are not limited to, mineral or organic acid salts of basic residues such as amines; alkali or organic salts of acidic residues such as carboxylic acids; and the like.
• Prodrugs are considered to be any covalently bonded carriers which release the active parent drug according to formula (I) in vivo when such prodrug is administered to a mammalian subject.
• Prodrugs of the compounds of formula (I) are prepared by modifying functional groups present in the compounds in such a way that the modifications are cleaved, either in routine manipulation or in vivo, to the parent compounds.
• Prodrugs include compounds of formula (I) wherein hydroxy, amine, or sulfhydryl groups are bonded to any group that, when administered to a mammalian subject, cleaves to form a free hydroxyi, 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.
• compositions of the compounds of the invention can be prepared by reacting the free acid or base forms of these compounds with a stoichiometric amount of the appropriate base or acid in water or in an organic solvent, or in a mixture of the two; generally, nonaqueous media like ether, ethyl acetate, ethanol, isopropanol, or acetonitrile are preferred. Lists of suitable salts are found in Remington's Pharmaceu ical Sciences. 17th ed.. Mack Publishing Company, Easton, PA, 1985, p. 1418, the disclosure of which is hereby incorporated by reference.
• amino acid as used herein means an organic compound containing both a basic amino group and an acidic carboxyl group. Included within this term are modified and unusual amino acids, such as those disclosed in, for example, Roberts and Vellaccio (1983) The Peptides. 5: 342-429, the teaching of which is hereby incorporated by reference.
• Modified or unusual amino acids which can be used to practice the invention include, but are not limited to, D-amino acids, hydroxylysine, 4-hydroxyproline, ornithine, 2, 4-diaminobutyric acid, homoarginine, norleucine, N-methylaminobutyric acid, naphthylalanine, phenylglycine, ⁇ -phenylproline, tert-leucine, 4-aminocyclohexylalanine, N-methyl-norleucine,
• amino acid residue means that portion of an amino acid (as defined herein) that is present in a peptide.
• peptide as used herein means a linear compound that consists of two or more amino acids (as defined herein) that are linked by means of a peptide bond.
• peptide also includes compounds containing both peptide and non-peptide components, such as pseudopeptide or peptide mimetic residues or other non-amino acid components. Such a compound containing both peptide and non-peptide components may also be referred to as a "peptide analog".
• a “pseudopeptide” or “peptide mimetic” is a compound which mimics the structure of an amino acid residue or a peptide, for example, by using linking groups other than amide linkages between the peptide mimetic and an amino acid residue (pseudopeptide bonds) and/or by using non-amino acid substituents and/or a modified amino acid residue.
• a “pseudopeptide residue” means that portion of an pseudopeptide or peptide mimetic (as defined herein) that is present in a peptide.
• peptide bond means a covalent amide linkage formed by loss of a molecule of water between the carboxyl group of one amino acid and the amino group of a second amino acid.
• peptide bonds includes peptide bond isosteres which may be used in place of or as substitutes for the normal amide linkage. These substitute or amide "equivalent” linkages are formed from combinations of atoms not normally found in 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.
• NMeAmf N-Methylaminomethylphenylalanine NMeAsp ⁇ -N-methyl aspartic acid NMeGly or MeGly N-methyl glycine NMe-Mamb N-methyl-3-aminomethylbenzoic acid
• Phg phenylglycine
• Trp tryptophan
• peptides are elongated by deprotecting the ⁇ -amine of the C-terminal residue and coupling the next suitably protected amino acid through a peptide linkage using the methods described. This deprotection and coupling procedure is repeated until the desired sequence is obtained.
• This coupling can be performed with the constituent amino acids in a stepwise fashion, or condensation of fragments (two to several amino acids), or combination of both processes, or by solid phase peptide synthesis according to the method originally described by Merrifield, J. Am. Chem. Soc, 85, 2149-2154 (1963), the disclosure of which is hereby incorporated by reference.
• the compounds of the invention may also be synthesized using automated peptide synthesizing equipment.
• procedures for peptide synthesis are described in Stewart and Young, "Solid Phase Peptide Synthesis", 2nd ed, Pierce Chemical Co., Rockford, IL (1984) ; Gross, Meienhofer, Udenfriend, Eds., "The Peptides: Analysis, Synthesis, Biology, Vol. 1, 2, 3, 5, and 9, Academic Press, New York, (1980- 1987); Bodanszky, “Peptide Chemistry: A Practical Textbook", Springer-Verlag, New York (1988); and Bodanszky et al. "The Practice of Peptide Sythesis” Springer-Verlag, New York (1984), the disclosures of which are hereby incorporated by reference .
• the coupling between two amino acid derivatives, an amino acid and a peptide, two peptide fragments, or the cyclization of a peptide can be carried out using standard coupling procedures such as the azide method, mixed carbonic acid anhydride (isobutyl chloroformate) method, carbodiimide (dicyclohexylcarbodiimide, diisopropylcarbodiimide, or water-soluble carbodiimides) method, active ester (p-nitrophenyl ester, N- hydroxysuccinic imido ester) method, Woodward reagent K method, carbonyldiimidazole method, phosphorus reagents such as BOP-C1, 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 solution (liquid phase) or solid phase.
• the functional groups of the constituent amino acids must be protected during the coupling reactions to avoid undesired bonds being formed.
• the protecting groups that can be used are listed in Greene, "Protective Groups in Organic Synthesis” John Wiley & Sons, New York (1981) and "The Peptides: Analysis, Sythesis, Biology, Vol. 3, Academic Press, New York (1981), the disclosure of which is hereby incorporated by reference.
• the ⁇ -carboxyl group of the C-terminal residue is usually protected by an ester that can be cleaved to give the carboxylic acid.
• These protecting groups include: 1) alkyl esters such as methyl and t-butyl, 2) aryl esters such as benzyl and substituted benzyl, or 3) esters which can be cleaved by mild base treatment or mild reductive means such as trichloroethyl and phenacyl esters.
• the C-terminal amino acid is attached to an insoluble carrier (usually polystyrene) .
• insoluble carriers contain a group which will react with the carboxyl group to form a bond which is stable to the elongation conditions but readily cleaved later. Examples of which are: oxime resin (DeGrado and Kaiser (1980) J. Org . Chem . 45, 1295-1300) chloro or bromomethyl resin, hydroxymethyl resin, and aminomethyl resin. Many of these resins are commercially available with the desired C-terminal amino acid already incorporated.
• acyl types such as formyl, trifluoroacetyl, phthalyl, and p- toluenesulfonyl
• aromatic carbamate types such as benzyloxycarbonyl (Cbz) and substituted benzyloxycarbonyls, 1- (p-biphenyl)-1- methylethoxycarbonyl, and 9-fluorenylmethyloxycarbonyl (Fmoc)
• 3) aliphatic carbamate types such as tert- butyloxycarbonyl (Boc) , ethoxycarbonyl, diisopropylmethoxycarbonyl, and allyloxycarbonyl
• cyclic alkyl carbamate types such as cyclopentyloxycarbonyl and adamantyloxycarbonyl
• 5) alkyl types such as triphen
• the ⁇ -amino protecting group is cleaved prior to the coupling of the next amino acid.
• the methods of choice are trifluoroacetic acid, neat or in dichioromethane, or HCl in dioxane.
• the resulting ammonium salt is then neutralized either prior to the coupling or in situ with basic solutions such as aqueous buffers, or tertiary amines in dichioromethane or dimethylformamide.
• 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 .
• Any of the amino acids bearing side chain functionalities must be protected during the preparation of the peptide using any of the above-identified groups.
• Those skilled in the art will appreciate that the selection and use of appropriate protecting groups for these side chain functionalities will depend upon the amino acid and presence of other protecting groups in the peptide. The selection of such a protecting group is important in that it must not be removed during the deprotection and coupling of the ⁇ -amino group.
• Boc when Boc is chosen for the ⁇ -amine protection the following protecting groups are acceptable: p-toluenesulfonyl (tosyl) moieties and nitro for arginine; benzyloxycarbonyl, substituted benzyloxycarbonyls, or tosyl for lysine; benzyl or alkyl esters such as cyclopentyl for glutamic and aspartic acids; benzyl ethers for serine and threonine; benzyl ethers, substituted benzyl ethers or 2- bromobenzyloxycarbonyl for tyrosine; p-methylbenzyl, p- methoxybenzyl, acetamidomethyl, benzyl, or t- butylsulfonyl for cysteine; and the indole of tryptophan can either be left unprotected or protected with a formyl group.
• tert-butyl based protecting groups are acceptable.
• Boc can be used for lysine, tert-butyl ether for serine, threonine and tyrosine, and tert-butyl ester for glutamic and aspartic acids.
• 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.
• the peptide should be removed from the resin without simultaneously removing protecting groups from functional groups that might interfere with the cyclization process.
• the cleavage conditions need to be chosen such that a free ⁇ - carboxylate and a free ⁇ -amino group are generated without simultaneously removing other protecting groups.
• the peptide may be removed from the resin by hydrazinolysis, and then coupled by the azide method.
• Another very convenient method involves the synthesis of peptides on an oxime resin, followed by intramolecular nucleophilic displacement from the resin, which generates a cyclic peptide (Osapay, Profit, and Taylor (1990) Tetrahedron Letters 43, 6121-6124) .
• the oxime resin is employed, the Boc protection scheme is generally chosen.
• the preferred method for removing side chain protecting groups generally involves treatment with anhydrous HF containing additives such as dimethyl sulfide, anisole, thioanisole, or p-cresol at 0 °C.
• the cleavage of the peptide can also be accomplished by other acid reagents such as trifluoromethanesulfonic acid/trifluoroacetic acid mixtures .
• Unusual amino acids used in this invention can be synthesized by standard methods familiar to those skilled in the art ("The Peptides: Analysis, Sythesis, Biology, Vol. 5, pp. 342-449, Academic Press, New York (1981)) .
• N-Alkyl amino acids can be prepared using procedures described in previously (Cheung et al . , (1977) Can . J. Chem. 55, 906; Freidinger et al . , (1982) J. Org. Chem . 48, 77 (1982)), which are incorporated here by reference.
• the compounds of the present invention may be prepared using the procedures further detailed below as well as the procedures described in PCT Patent Application International Publication Number WO 93/07170, the disclosure of which is hereby incorporated herein by reference . Those compounds referred to by Example Number which are not detailed herein are disclosed in detail in PCT International Publication Number WO 93/07170.
• HBTU 2- (lH-Benzotriazol-1-yl)-1,1,3, 3-tetramethyluronium hexafluorophosphate
• TBTU 2-(lH-Benzotriazol-1-yl)-1,1,3, 3-tetramethyluronium hexafluorophosphate
• NMM N-methylmorpholine
• Abu D-2-aminobutyric acid
• DIEA diisopropylethylamine
• Palladium on carbon catalyst (10% Pd) was purchased from Fluka Chemical Company. Absolute ethanol was obtained from Quantum Chemical Corporation.
• Thin layer chromatography (TLC) was performed on Silica Gel 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 spray. Melting points were determined using a Thomas Hoover or Electrothermal 9200 melting point apparatus and are uncorrected. HPLC analyses were performed on either a Hewlett Packard 1090, Waters Delta Prep 3000, Rainin, or DuPont 8800 system.
• NMR spectra were recorded on a 300 MHz General Electric QE-300, Varian 300, or Varian 400 spectrometer.
• FAB-MS Fast atom bombardment mass spectrometry
• Boc-aminomethylbenzoic acid derivatives useful as intermediates in the synthesis of the compounds of the invention are prepared using standard procedures, for example, as described in Tett.
• 3-Cyanobenzoic acid (10.0 g, 68 mmol) was dissolved in 200 ml ethanol by heating in a 35-50 C water bath. Concentrated HCl (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 under an atmosphere of hydrogen for 38 hours, filtered through a scintered glass funnel, and washed thoroughly with H2O. The ethanol was removed under reduced pressure and the remaining aqueous layer, which contained a white solid, was diluted to 250 ml with additional H2O.
• the aqueous layer was then acidified to pH 2 with 2N HCl and then 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.
• the title compound can be prepared according to standard procedures, for examples, as disclosed in Olsen, J. Org. Chem. (1970) 35: 1912), and as shown schematically below.
• Amine hydrochloride (0.40 g, 2 mmol) was dissolved in 15 ml water. A solution of BOC-ON (0.52 g, 2.1 mmol) in 15 ml acetone was added, followed by the addition of triethylamine (0.8 ml, 6 mmol) . Reaction was allowed to proceed for 20 h. Reaction mixture was concentrated, partitioned between ethyl acetate and water. Aqueous layer was acidified to pH 2 using 10% HCl solution.
• t-Butyloxycarbonyl-3-aminophenylacetic acids useful as intermediates in the synthesis of the compounds of the invention are prepared using standard procedures, for example, as described in Collman and Groh (1982) J. . m. Chem . Soc , 104: 1391, and as shown schematically below.
• 2-Aminomethylbenzoic acid-HCl and 2- aminomethylphenylacetic acid-HCl useful as intermediates in the synthesis of the compounds of the invention are prepared using standard procedures, for example, as described in Naito et al J. Antibiotics, 30: 698 (1977); or Young and Sweet J. Am. Chem. Soc , 80: 800 (1958), and as shown schematically below.
• Mamb useful as carbocylic residues R 31 in the cyclic peptides of the invention include aminoalkyl- naphthoic acid and aminoalkyl-tetrahydronaphthoic acid residues.
• Representative aminoalkyl-naphthoic acid and aminoalkyl-tetrahydronaphthoic acid intermediates useful in the synthesis of cyclic peptides of the present invention are described below. The synthesis of these intermediates is outlined below in Scheme 4a.
• 4-phenylbutyric acid (1) was converted to the ethyl ester (2) which was acylated via aluminum chloride and acetylchloride to give 4-acetylphenylbutyric acid ethyl ester (3) .
• This ester was subjected to saponification to give 4- acetylphenylbutyric acid (4) .
• the acetyl 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.
• the tetralone was split into two portions and some was converted to the oxime (7) using sodium acetate and hydroxylamine hydrochloride.
• the oxime was subjected to hydrogenolysis to give the racemic mixture of 8-amino- 5, 6, 7, 8-tetrahydro-2-naphthoic acid as the hydrochloride (8) for use as an intermediate for incorporation into the cyclic peptide.
• 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 dichioromethane. The combined organic layers were backwashed with dilute hydrochloric acid and water, dried over anhydrous magnesium sulfate and evaporated to dryness under reduced pressure.
• 4-carboxyphenylbutyric acid 10.40 g, 0.05 mol
• aluminum chloride 33.34 g, 0.25 mol
• sodium chloride 2.90 g, 0.05 mol
• 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) .
• 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) 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 remaining tetralone was then converted to the methyl ester (9) .
• the tetralone methyl ester (9) was converted, first, to the cyanohydrin by treatment with trimethylsilylcyanide and zinc iodide and then, via the in situ dehydration with phosphorous oxychloride in pyridine, to the methyl 8-cyano-5, 6-dihydro-2-naphthoate (11) .
• This naphthoate was divided into two portions and some was subjected to hydrogenolysis, N-BOC-protection and saponification to give N- (BOC) -8-aminomethyl- 5, 6, 7, 8-tetrahydro-2-naphthoic acid (12) as an intermediate for incorporation into the cyclic peptide.
• Part A A mixture of l-tetralon-7-carboxylic acid (7.0 g, 0.037 mol) in methanol (13.6 mL, 10.8 g, 0.30 mol) with a catalytic amount of hydrochloriic acid (0.07 mL, 0.12 g, 0.0012 mol) was stirred at reflux over 5 hours. The cooled reaction mixture was poured into ice water and extracted with ethyl acetate. The combined organic layers were backwashed with water and brine, dried over anhydrous magnesium sulfate and evaporated to dryness under reduced pressure. The resulting solid was purified by flash chromatography using hexane:ethyl acetate: :75 :25.
• 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 5 (20 mL) was stirred at ambient temperature over 15 hours. Then added, sequentially and dropwise, was pyridine (20 mL) and phosphorous oxychloride (4.0 mL, 6.55 g, 0.0425 mol) . The reaction mixture was stirred at reflux over 1 hour then evaporated to dryness under • 10 reduced pressure.
• Part C A mixture of methyl 8-cyano-5, 6-dihydro-2- 20 naphthoate (0.80 g, 0.0038 mol) in methanol (25 L) 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 25 over Celite and washed with methanol.
• Part D A solution of methyl 8-aminomethyl-5, 6, 7, 8- 35 tetrahydro-2-naphthoate (0.78 g, 0.0036 mol) and triethylamine (0.55 mL, 0.40 g, 0.004 mol) in aqueous tetrahydrofuran (50%, 75 mL) was added, portionwise as a solid, 2- (teri-butoxycarbonyloxyimino) -2- phenylacetonitrile (0.99 g, 0.004 mol) . All was stirred at ambient temperature over 3 hours. The solution was concentrated to half volume and extracted with diethylether.
• the aqueous layer was then acidified to a pH of 1.0 using hydrochloric acid (IN) and then extraced with ethyl acetate.
• the combined organic layers were dried over anhydrous magnesium sulfate and evaporated to dryness under reduced pressure.
• the remaining naphthoate (11) was treated with 2,3- dichloro-5, 6-dicyano-l, 4-benzoquinone (DDQ) in dioxane to aromatize the adjacent ring to give the methyl 8- cyano-2-naphthoate (13) .
• DDQ 2,3- dichloro-5, 6-dicyano-l, 4-benzoquinone
• the nitrile was reduced via hydrogentation and the methyl ester saponified to the carboxylic acid.
• This acid was then N-BOC-protected to give N- (BOC)-8-aminomethyl-2-naphthoic acid (14) as an intermediate for incorporation into the cyclic peptide.
• Part B A mixture of methyl 8-cyano-2-naphthoate (1.0 g, 0.0047 mol) in methanol (35 mL) with concentrated hydrochloric acid (0.69 mL) andpalladium on carbon catalyst (0.20 g, 5% Pd/C) was shaken for 6 hours at ambient temperature under anatmosphere of hydrogen (50 psi) .
• the reaction mixture was filtered over Celite e and washed with methanol. The filtrate was evaporated to dryness under reduced pressure and the residue was triturated with hexane to give methyl 8-aminomethyl-2- naphthoate (0.76 g, 0.0035 mol, 75%) as an oil.
• 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 L) 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.
• 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 substitution level is then determined using either the picric acid test (Sarin, Kent, Tarn, and Merrifield, (1981) Anal . Biochem .
• NMM NMM (based on the amount of amino acid used) and the coupling times range from 1 hour to several days. The completeness of coupling is monitored by qualitative ninhydrin assay, or picric acid assay in cases where the amino acid was coupled to a secondary amine. Amino acids are recoupled if necessary based on these results.
• the N- terminal Boc group is removed by treatment with 25% TFA in DCM for 30 minutes. The resin is then neutralized by treatment with 10% DIEA in DCM. Cyclization with concomitant cleavage of the peptide is accomplished using the method of Osapay and Taylor ((1990) J. Am . Chem .
• Soc 112, 6046
• HOAc Hydrophilic acid
• the DMF filtrate is evaporated, redissolved in HOAc or 1:1 acetonitrile: H2O, and lyophilized to obtain protected, cyclized material.
• the material may be dissolved in methanol and precipitated with ether to obtain the protected, cyclized material. This is then treated using standard procedures with anhydrous hydrogen fluoride (Stewart and Young (1984) "Solid Phase Peptide Synthesis", 2nd. edition.
• the crude product may be purified by reversed-phase HPLC using a 2.5 cm preparative Vydac C18 column with a linear acetonitrile gradient containing 0.1% TFA to produce pure cyclized material.
• Boc-Arg(Tos) Boc-N-a-MeArg (Tos) , Boc- Gly, Boc-Asp (OcHex) , Boc-3-aminomethyl-4-iodo-benzoic acid, Boc-D-Ile, Boc-NMeAsp(OcHex) , Boc-NMe-Mamb, Boc-D- Phg, Boc-D-Asp (OBzl) , Boc-L-Asp(OcHex) , Boc-aMe- Asp (OcHex), Boc-bMe-Asp (OcHex) , Boc-L-Ala, Boc-L-Pro, Boc-D-Nle, Boc-D-Leu, Boc-D-Val, Boc-D-2-aminobutyric acid (Boc-D-Abu) , Boc-Phe, Boc-D-Ser (Bzl
• the title compound was prepared using the general procedure described for cyclo- (D-Val-NMeArg-Gly-Asp- Ma b) (Example 4) .
• the DCC/DMAP method was used for attachment of Boc-Mamb to the oxime resin.
• TBTU was used as the coupling reagent.
• the peptide was prepared on a 0.596 mmol scale to give the protected cyclic peptide (182 mg,38.4%) .
• the peptide (176 mg) and 0.176 L of anisole were treated with anhydrous hydrogen o fluoride at 0 C for 20 minutes.
• Example 4 The zwitterion of Example 4 was converted to the mesyl (methanesulfonate) salt of Example 4 (Example 4 (methane-sulfonate) ) by refiuxing the zwitterion with stirring in isopropanol at 25 mg/ml and slowly adding a solution of 1.0 molar equivalent methanesulfonie 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 rinsed three times with cold isopropanol and dried under vacuum to constant weight.
• Example 4 ( ⁇ -naphthalenesulfonate) : zwitterion + 1.0 equiv. ⁇ -naphthalenesulfonic acid.
• Example 4 ( ⁇ -naphthalenesulfonate) : zwitterion + 1.0 equiv. ⁇ -naphthalenesulfonic acid.
• Example 4 (benzenesulfonate) : zwitterion + 1.0 equiv. benezene-sulfonic acid.
• Example 4 (p-toluenesulfonate) : zwitterion + 1.0 equiv. p-toluene-sulfonic acid.
• the following salts of the compound of Example 4 were prepared by crystallization of the compound from aqueous systems.
• Example 4 10 mg amorphous Example 4 (made by lyophilizing the zwitterion from a solution of 2 molar equivalents of acetic acid in water) dissolved per ml 1 N H2SO4, pH adjusted to 2.5. On standing at room temperature, a precipitate formed. This was filtered through a sintered glass funnel and dried under vacuum to constant weight .
• Example 4 (methanesulfonate (mesyl)) : 100 mg amorphous DMP728 dissolved per ml water + 1.2 molar equiv. methanesulfonie acid (this was obtained as a 4M aqueous solution) . On standing at room temperature, a large flat crystal was formed.
• Example 4 (p-toluenesulfonate) : 100 mg zwitterion dissolved per ml water + 1.2 molar equiv. toluenesulfonic acid added. On standing at room temperature, a precipitate formed. This was filtered through a sintered glass funnel and dried under vacuum to constant weight.
• the title compound was prepared using the general procedure described for cyclo- (D-Val-NMeArg-Gly-Asp- Mamb) (Example 4) .
• the DCC/DMAP method was used for the attachment of Boc-Mamb to the resin.
• the peptide was prepared on a 0.179 mmol scale to give the protected cyclic peptide (105 mg, 69.7%) .
• the peptide (105 mg) and 0.105 mL of anisole were treated with anhydrous o hydrogen fluoride at 0 C for 20 minutes.
• the title compound was prepared using the general procedure described for cyclo- (D-Val-NMeArg-Gly-Asp- Mamb) (example 4) .
• the DCC/DMAP method was used for attachment of Boc-Mamb to the oxime resin.
• TBTU was used as the coupling reagent.
• the peptide was prepared on a 0.596 mmol scale to give the protected cyclic peptide (273 mg, 57.6%) .
• the peptide (263 mg) and 0.263 mL of anisole were treated with anhydrous hydrogen fluoride at O C for 20 minutes.
• R 1 H
• R 2 H
• the title compound was prepared using the general procedure described for cyclo- (D-Val-NMeArg-Gly-Asp- Mamb) (example 4) .
• the DCC/DMAP method was used for attachment of Boc-Mamb to the oxime resin.
• TBTU was used as the coupling reagent.
• the peptide was prepared on a 0.596 mmol scale to give the protected cyclic peptide (241 mg, 50.8%) .
• the peptide (235 mg) and 0.235 mL of anisole were treated with anhydrous hydrogen o fluoride at 0 C for 20 minutes.
• R 1 H
• R 2 H
• 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- NMe-Lysine amino acid was obtained as a side product during the synthesis of the corresponding di-NMe-lysine derivative.
• the title compound was prepare by the general solution-phase procedure described above for cyclo- (D- Val-NMeArg-Gly-Asp-Mamb) , except that 4,4'- dinitrobenzophenone oxime was employed.
• the cyclic peptide (330 mg, 0.40 mmol) was deprotected with excess HF in the presence of anisole as scavenger.
• the title compound was prepare by the general solution-phase procedure described above for cyclo- (D- Va1-NMeArg-Gly-Asp-Mamb) , except that 4,4'- dinitrobenzophenone oxime was employed.
• the cyclic peptide 350 mg, 0.38 mmol
• Purification was accomplished by reversed-phase HPLC on a preparative Vydac C18 column (2.5 cm) using a 1.0% / minute gradient of 10 to 38% acetonitrile containing 0.1% trifluoroacetic acid to give the TFA salt of the title compound (150 mg, 49%) as a fluffy white solid; 1 H NMR
• the title compound was prepare by the general solution-phase procedure described above for cyclo- (D- Val-NMeArg-Gly-Asp-Mamb) , except that 4,4'- dinitrobenzophenone oxime was employed.
• the cyclic peptide 130 mg, 0.16 mmol was deprotected with excess HF in the presence of anisole as scavenger.
• Step 1 N g -benzyloxvcarbonyl-N-g-methyl-4-cyano-L-2- aminobutvric acid
• the dried oily product was taken up in 300 mL dry THF and 49.8 mL (800 mmol) methyl iodide in a flask bottle protected from moisture and the solution was cooled in an ice bath. To it was slowly added 10 g sodium hydride (250 mmol, 60% dispersion in oil) . The mixture was stirred in the ice bath for 1 h and then at room temperature for 22 h. Ethyl acetate (50 mL) was added, and after stirring for 10 min, 100 mL water was added slowly. The solution was acidified with a few drops of 4 N HCl to pH8-9 and then concentrated at 30° C to remove the organic solvents.
• Step 2 N*a-methy]-4-cyano-L-2-aminob ⁇ tvric acid-N- carboxyanhydride
• Step 3 N-Boc-D- -aminoh._t.yryl -Nfl-methvl-4-cvano-L-2- am-inoh.-tyryl- ⁇ lvcine t-b__tvl ester
• glycine t-butyl ester hydrochloride 3.68 g, 22 mmol
• example 2 3.36 g, 20 mmol
• dry acetonitrile was stirred at -20° C for 1 h, and the solvent was reduced to about 10 mL.
• N-Boc-D-2-aminobutyric acid dicyclohexylamine salt 8.08 g, 21 mmol
• diphenylphosphinic chloride 3.91 mL, 20.5 mmol
• 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.
• Step 4 N-Boc-D-2-aminobutyryl-N g -methyl-N.a.—Nffll- (bisbenzyloxycarbonyl)-L-ar ⁇ inyl- ⁇ lycine t-butyl ester
• the compound of Step 3 (4.63 g, 10.5 mmol) was dissolved in 70 mL methanol in a Parr bottle and to it was added a cold solution of 1.2 mL concentrated hydrochloric acid (38%) in 10 mL methanol followed by 200 mg platinum(IV) oxide. The mixture was hydrogenated at 55 psi for 1 h, the catalyst was filtered off, and 2.09 mL (15 mmol) triethylamine was added.
• Step 5 D-2-am-inon__tyryl-N-a-methyl-N*a. N*al- (bisbenzyloxycarbonyl) -L-arginyl-glycine TFA salt
• Step 7 Fmoc-L-asoartvl (t-butvl)-3- (aminomethyl) - benzoic acid
• step 8 Fmoc-L-aspartyl(t-fr ⁇ tyl)-3- (aminomethyl)benzoyl-D-2-aminobutvrvl-N*a-methvl-NH. N***-*— (bisbenzvloxycarbonvl)-L-ar ⁇ invl- ⁇ lvcine
• Step 9 Cvclo.L-aspartvl (t-butvl)-3- (aminomethyl)benzovl-D-2-aminobutvrvl-Nfl. N- ⁇ l-
• Step 10 Cvclo.L-aspartvl-3- (aminomethyl)benzovl-D-2- aminobutvrvl-N-H. N-ttl- (bisbenzyloxvcarbonvl) -1,-ar ⁇ invl- q-T-ycy l
• Patent Applications which are filed on the same day as the present application: Attorney Docket No. DM-6535, named inventors Maduskuie and Pes i; Attorney Docket No. DM-6650, named inventors Zhang, Ma, and De Grado; and Attorney Docket No. DM-6665, named inventors De Grado, Dorow, Ward, and Xue.
• the compounds of this invention possess antiplatelet efficacy, as evidenced by their activity in standard platelet aggregation assays or platelet fibrinogen binding assays, as described below.
• a compound is considered to be active in these assays if it has an 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 citrated Vacutainer tubes. The blood was centrifuged for 15 minutes at 150 x g at room temperature, and platelet-rich plasma (PRP) was removed. The remaining blood was centrifuged for 15 minutes at 1500 x g at room temperature, and platelet-poor plasma (PPP) was removed. Samples were assayed on a aggregometer (PAP-4 Platelet Aggregation Profiler), using PPP as the blank (100% transmittance) . 200 ⁇ l of PRP was added to each micro test tube, and transmittance was set to 0%.
• PAP-4 Platelet Aggregation Profiler PPP as the blank (100% transmittance
• Platelet-Fibrinogen Binding Assay Binding of ⁇ I-fibrinogen to platelets was performed as described by Bennett et al. (1983) Proc. Natl. Acad. Sci . USA 80: 2417-2422, with some modifications as described below. Human PRP (h-PRP) was applied to a Sepharose column for the purification of platelet fractions. Aliquots of platelets (5 X 10 8 cells) along with 1 mM calcium chloride were added to removable 96 well plates prior to the activation of the human gel purified platelets (h- GPP) .
• Activation of the human gel purified platelets was achieved using ADP, collagen, arachidonate, epinephrine, and/or thrombin in the presence of the ligand, 1 ⁇ -fibrinogen.
• the 2 ⁇ I-fibrinogen bound to the activated, platelets was separated from the free form by centrifugation and then counted on a gamma counter.
• the test compounds were added at various concentrations prior to the activation of the platelets.
• 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 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 compound capable of achieving 50% clot lysis) of less than about 1 mM, more preferably an IC 5 0 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 an IC 5 0 value of about 0.0005 mM.
• an IC50 value that is, the molar concentration of the cyclic compound capable of achieving 50% clot lysis
• IC 50 determinations may be made using a standard thrombolysis assay, as described below.
• Another class of preferred thrombolytic compounds of the invention include those compounds which have a Kd of ⁇ 100 nM, preferably ⁇ 10 nM, most preferably 0.1 to 1.0 nM.
• Thrombolytic 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, and placed into 10 ml citrated Vacutainer tubes. The blood was centrifuged for 15 minutes at 1500 x g at room temperature, and platelet rich plasma (PRP) was removed. To the PRP was then added 1 x 10 ⁇ 3 M of the agonist ADP, epinephrine, collagen, arachidonate, serotonin or thrombin, or a mixture thereof, and the PRP incubated for 30 minutes. The PRP was centrifuged for 12 minutes at 2500 x g at room temperature.
• PRP platelet rich plasma
• test compounds were added at various concentrations. Test samples were taken at various time points and the platelets were counted using the Coulter Counter. To determine the percent of lysis, the platelet count at a time point subsequent to the addition of the test compound was subtracted from the platelet count at the zero time point, and the resulting number divided by the platelet count at the zero time point. Multiplying this result by 100 yielded the percentage of clot lysis achieved by the test compound. For the IC50 evaluation, the test compounds were added at various concentrations, and the percentage of lysis caused by the test compounds was calculated.
• novel cyclic compounds of the invention are also useful in combination products, that is, in pharmaceutical compositions containing the novel cyclic compounds of the invention in combination with anti-coagulant agents such as warfarin or heparin, or antiplatelet agents such as aspirin, piroxicam or ticlopidine, or thrombin inhibitors such as boropeptides, hirudin or argatroban, or thrombolytic agents such as tissue plasminogen activator, anistreplase, urokinase or streptokinase, or combinations thereof.
• anti-coagulant agents such as warfarin or heparin
• antiplatelet agents such as aspirin, piroxicam or ticlopidine
• thrombin inhibitors such as boropeptides, hirudin or argatroban
• thrombolytic agents such as tissue plasminogen activator, anistreplase, urokinase or streptokinase, or combinations thereof.
• Compound A was shown not only to inhibit platelet aggregation induced by agonists, but also to deaggregate platelets after the initiation of aggregation.
• the deaggregation efficacy of Compound A was dependent on its concentration and the time of addition post-initiation of platelet activation. The earlier the addition of Compound A after the induction of aggregation, the greater its deaggregatory efficacy (Fig. la) .
• the cyclic glycoprotein Ilb/III antagonist compounds of this invention have also been shown to displace 125 I-fibrinogen bound to activated platelets in a platelet-fibrinogen binding assay similar to the platelet-fibrinogen binding assay previously described.
• cyclic compounds of the present invention namely the compound of Example 3 (cyclo- (D- Abu-NMeArg-Gly-Asp-Mamb; the compound of formula (II) wherein R 1 and R 2 are H, J is D-2-aminobutyric acid, K is ⁇ -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 compound of formula (II) wherein R 1 and R 2 are H, J is D-valine, K is ⁇ -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 on the reversal of the aggregatory response (deaggregation) to 10
• representative compounds of the present invention are shown not only to inhibit platelet aggregation induced by agonists, but also to deaggregate platelets after the initiation of aggregation ( Figure la) .
• the deaggregation efficacy of compound A was dependent on its concentration and the time of addition post-initiation of platelet activation. The earlier the addition of Compound A after the induction of aggregation, the greater its deaggregatory efficacy.
• Figure lb shows the lytic effect of Compound A and B on an already formed platelet rich clot.
• the clot was formed by incubating platelets with a mixture of agonists (TEAC mixture) , which consists of thrombin (0.01 U/ml) , epinephrine (250 uM) , ADP (250 uM) , and collagen (10 ug.ml), for 30 minutes.
• TEAC mixture a mixture of agonists
• a linear peptide of sequence arginine-glycine-aspartic acid-serine (RGDS) was also tested in the thrombolytic assay.
• Compounds A and B was tested at a concentration of 0.001 mM using the thrombolytic assay described above, with platelet stimulation being carried out using 1 x 10 ⁇ 3 M concentration of ADP.
• the standard thrombolytics tissue plasminogen activator (tPA; 10 ⁇ g/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.
• the compounds of the invention demonstrated a significant effect on the lysis of an already formed platelet-rich clot, with Compound A providing significantly better clot lysis than tissue plasminogen activator, urokinase, and streptokinase, and Compound B providing significantly better clot lysis than streptokinase.
• Compound A had an excellent lysis percentage of 70% or greater.
• Figure III shows the effect of 1 uM of Compound A on the lysis of an already formed platelet-rich clot.
• the clot was formed by the addition of TEAC mixture (which consists of thrombin (0.01 U/ml), epinephrine (250 uM) , ADP (250 uM) , and collagen (10 ug.ml)) for 30 minutes.
• Compound A resulted in significant clot lysis by itself as compared to tissue plasminogen activator (tPA; 10 ⁇ g/ml), urokinase (UK; 900 units/ml) and streptokinase (SK; 500 units/ml) .
• tPA tissue plasminogen activator
• urokinase UK
• streptokinase streptokinase
• Compound C was tested at a concentration of 1 uM using the thrombolytic assay described above, both alone and in combination with the standard thrombolytics tissue plasminogen activator (tPA; 10 ⁇ g/ml) , urokinase (UK; 900 units/ml) and streptokinase (SK; 500 units/ml) .
• tissue plasminogen activator tPA
• urokinase UK
• streptokinase streptokinase
• Compound D was tested at a concentration of 1 uM using the thrombolytic assay described above, both alone and in combination with the standard thrombolytics tissue plasminogen activator (tPA; 10 ⁇ g/ml), urokinase (UK; 900 units/ml) and streptokinase (SK; 500 units/ml) .
• tissue plasminogen activator tPA
• urokinase UK
• streptokinase streptokinase
• N 3 in each group.
• Antiplatelet combination of the cyclic GPTTb/TIIa receptor antagonist of the present invention and aspirin and/or heparin is provided.
• Treatment Groups Group I (Saline) : I.V. bolus of saline.
• Group II (Aspirin) : 10 mg/kg, po - 30 min prior to blood sampling.
• Group III (Compound A) : 0.08 mg/kg, I.V. bolus.
• Group IV (Aspirin/Compound A) : Aspirin at 10 mg/kg, po - 30 min prior to the administration of
• the compounds of this invention can be administered by any means that produces contact of the active agent with the agent's site of action, glycoprotein Ilb/IIIa (GPIIb/IIIa), in the body of a mammal. They can be administered by any conventional means available for use in conjunction with pharmaceuticals, either as individual therapeutic agents or in a combination of therapeutic agents, such as a second antiplatelet agent such as aspirin, piroxicam, or ticlopidine which are agonist-specific, or an anti-coagulant such as warfarin or heparin, or a thrombin inhibitor such as a boropeptide, hirudin or argatroban, or a thrombolytic agent such as tissue plasminogen activator, anistreplase, urokinase or streptokinase, or combinations thereof.
• the compounds of the invention, or compounds of the invention in combination with other therapeutic agents can be administered alone, but generally administered with a pharmaceutical carrier selected on the basis of the chosen route of administration and standard pharmaceutical practice.
• the dosage of the novel cyclic compounds of this invention administered will, of course, vary depending upon known factors, such as the pharmacodynamic characteristics of the particular agent and its mode and route of administration; the age, health and weight of the recipient; the nature and extent of the symptoms; the kind of concurrent treatment; the frequency of treatment; and the effect desired.
• a daily dosage of active ingredient can be expected to be about 0.01 to 10 milligrams per kilogram of body weight.
• Dosage forms contain from about 1 milligram to about 100 milligrams of active ingredient per unit .
• the active ingredient will ordinarily be present in an amount of about 0.5-95% by weight based on the total weight of the composition.
• the active ingredient can be administered orally in solid dosage forms, such as capsules, tablets, and powders, or in liquid dosage forms, such as elixirs, syrups, and suspensions. It can also be administered parenterally, in sterile liquid dosage forms.
• Gelatin capsules contain the active ingredient and powdered carriers, such as lactose, starch, cellulose derivatives, magnesium stearate, stearic acid, and the like. Similar diluents can be used to make compressed tablets. Both tablets and capsules can be manufactured as sustained release products to provide for continuous release of medication over a period of hours. Compressed tablets can be sugar coated or film coated to mask any unpleasant taste and protect the tablet from the atmosphere, or enteric coated for selective disintegration in the gastrointestinal tract.
• powdered carriers such as lactose, starch, cellulose derivatives, magnesium stearate, stearic acid, and the like. Similar diluents can be used to make compressed tablets. Both tablets and capsules can be manufactured as sustained release products to provide for continuous release of medication over a period of hours. Compressed tablets can be sugar coated or film coated to mask any unpleasant taste and protect the tablet from the atmosphere, or enteric coated for selective disintegration in the gastrointestinal tract.
• Liquid dosage forms for oral administration can contain coloring and flavoring to increase patient acceptance.
• water a suitable oil, saline, aqueous dextrose (glucose) , and related sugar solutions and glycols such as propylene glycol or polyethylene glycols are suitable carriers for parenteral solutions.
• saline aqueous dextrose (glucose)
• glycols such as propylene glycol or polyethylene glycols
• Solutions for parenteral administration preferably contain a water soluble salt of the active ingredient, suitable stabilizing agents, and if necessary, buffer substances.
• Antioxidizing agents such as sodium bisulfite, sodium sulfite, or ascorbic acid, either alone or combined, are suitable stabilizing agents.
• citric acid and its salts and sodium EDTA are suitable stabilizing agents.
• parenteral solutions can contain preservatives, such as benzalkonium chloride, methyl- or propyl-paraben, and chlorobutanol .
• Suitable pharmaceutical carriers are described in Remington's Pharmaceutical Sciences. Mack Publishing
• a large number of unit capsules are prepared by filling standard two-piece hard gelatin capsules each with 100 milligrams of powdered active ingredient, 150 milligrams of lactose, 50 milligrams of cellulose, and 6 milligrams magnesium stearate.
• Soft Gelatin Capsules A mixture of active ingredient in a digestable oil such as soybean oil, cottonseed oil or olive oil is prepared and injected by means of a positive displacement pump into gelatin to form soft gelatin capsules containing 100 milligrams of the active ingredient. The capsules are washed and dried.
• a digestable oil such as soybean oil, cottonseed oil or olive oil
• a large number of tablets are prepared by conventional procedures so that the dosage unit was 100 milligrams of active ingredient, 0.2 milligrams of colloidal silicon dioxide, 5 milligrams of magnesium stearate, 275 milligrams of microcrystalline cellulose, 11 milligrams of starch and 98.8 milligrams of lactose.
• Appropriate coatings may be applied to increase palatability or delay absorption.
• the combination products of this invention such as the novel cyclic Ilb/IIIa antagonist compounds of this invention in combination with an anti-coagulant agent such as warfarin or heparin, or an anti-platelet agent such as aspirin, piroxicam or ticlopidine, or a thrombin inhibitor such as a boropeptide, hirudin or argatroban, or a thrombolytic agent such as tissue plasminogen activator, anistreplase, urokinase or streptokinase, or combinations thereof, can be in any dosage form, such as those described above, and can also be administered in various ways, as described above.
• an anti-coagulant agent such as warfarin or heparin
• an anti-platelet agent such as aspirin, piroxicam or ticlopidine
• a thrombin inhibitor such as a boropeptide, hirudin or argatroban
• a thrombolytic agent such as tissue plasminogen activator, anistre
• the combination products of the invention are formulated together, in a single dosage form (that is, combined together in one capsule, tablet, powder, or liquid, etc.) .
• 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 agent, thrombin inhibitor, and/or thrombolytic agent may be administered at the same time (that is, together) , or in any order, for example the compounds of this invention are administered first, followed by administration of the anti-coagulant agent, anti-platelet agent, thrombin inhibitor, and/or thrombolytic agent .
• the administration of the compound of this invention and any anti-coagulant agent, anti-platelet agent, thrombin inhibitor, and/or thrombolytic agent occurs less than about one hour apart, more preferably less than about 30 minutes apart, even more preferably less than about 15 minutes apart, and most preferably less than about 5 minutes apart.
• administration of the combination products of the invention is oral.
• oral agent, oral inhibitor, oral compound, or the like, as used herein, denote compounds which may be orally administered.
• the cyclic Ilb/IIIa antagonist compounds of this invention and the anti-coagulant agent, anti-platelet agent, thrombin inhibitor, and/or thrombolytic agent are both administered in the same fashion (that is, for example, both orally) , if desired, they may each be administered in different fashions (that is, for example, one component of the combination product may be administered orally, and another component may be administered intravenously) .
• the dosage of the combination products of the invention may vary depending upon various factors such as the pharmacodynamic characteristics of the particular agent and its mode and route of administration, the age, health and weight of the recipient, the nature and extent of the symptoms, the kind of concurrent treatment, the frequency of treatment, and the effect desired, as described above.
• a daily 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 kilogram of patient body weight.
• the novel compounds of this invention generally may be present in an amount of about 5 to 10 milligrams, and the anti-coagulants in an amount of about 1 to 5 milligrams.
• a daily dosage may be about 0.01 to 25 milligrams of the cyclic compounds of this invention and about 50 to 150 milligrams of the additional anti-platelet agents, preferably about 0.1 to 1 milligrams of the novel compounds of this invention and about 1 to 3 milligrams of antiplatelet agents, per kilogram of patient body weight .
• the novel compounds of this invention may be present, for example, in an amount of about 5 milligrams, and the additional anti-platelet agent in an amount of about 150 milligrams, or, for example, in an amount of about 25 milligrams of the cyclic compound of this invention and about 50 milligrams of the additional antiplatelet agent.
• a daily dosage may be about 0.1 to 1 milligrams of the cyclic compound of this invention, per kilogram of patient body weight and, in the case of the thrombolytic agents, the usual dosage of the thrombolyic agent when administered alone may be reduced by about 70-80% when administered with a compound of the present invention.
• the novel compounds of this invention may be present, for example, in an amount of about 10 milligrams.
• the amount of each component in a typical daily dosage and typical dosage form may be reduced relative to the usual dosage of the agent when administered alone, in view of the additive or synergistic effect which would be obtained as a result of addition of further agents in accordance with the present invention.
• the potential exists for a chemical interaction between the combined active ingredients for example, a novel compound of this invention and an anti-coagulant such as warfarin or heparin, or a novel compound of this invention and an anti-platelet agent such as aspirin, piroxicam or ticlopidine, or a novel compound of this invention and a thrombin inhibitor such as a boropeptide, hirudin or argatroban, or a novel compound of this invention and a thrombolytic agent such as tissue plasminogen activator, anistreplase, urokinase or streptokinase, or combinations thereof) .
• the preferred dosage forms of the combination products of this invention are formulated such that although the active ingredients are combined in a single dosage form, the physical contact between the active ingredients is minimized (that is, reduced) .
• one embodiment of this invention where the product is orally administered provides for a combination product wherein one active ingredient is enteric coated.
• enteric coating one of the active ingredients it is possible not only to minimize the contact between the combined active ingredients, but also, it is possible to control the release of one of these components in the gastrointestinal tract such that one of these components is not released in the stomach but rather is released in the intestines.
• Another embodiment of this invention where oral administration is desired provides for a combination product wherein one of the active ingredients is coated with a sustained-release material which effects a sustained-release throughout the gastrointestinal tract and also serves to minimize physical contact between the combined active ingredients.
• the sustained-released component can be additionally enteric coated such that the release of this component occurs only in the intestine.
• Still another approach would involve the formulation of a combination product in which the one component is coated with a sustained and/or enteric release polymer, and the other component is also coated with a polymer such as a lowviscosity grade of hydroxypropyl methylcellulose (HPMC) or other appropriate materials as known in the art, in order to further separate the active components.
• HPMC hydroxypropyl methylcellulose
• the polymer coating serves to form an additional barrier to interaction with the other component.
• Dosage forms of the combination products of the present invention wherein one active ingredient is enteric coated can be in the form of tablets such that the enteric coated component and the other active ingredient are blended together and then compressed into a tablet or such that the enteric coated component is compressed into one tablet layer and the other active ingredient is compressed into an additional layer.
• one or more placebo layers may be present such that the placebo layer is between the layers of active ingredients.
• dosage forms of the present invention can be in the form of capsules wherein one active ingredient is compressed into a tablet or in the form of a plurality of microtablets, particles, granules or non-perils, which are then enteric coated. These enteric coated microtablets, particles, granules or non- perils are then placed into a capsule or compressed into a capsule along with a granulation of the other active ingredient.
• kits useful in, for example, the inhibition of platelet aggregation, the treatment of blood clots, and/or the treatment of thromboembolic 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 warfarin or heparin, or an antiplatelet agent such as aspirin, piroxicam or ticlopidine, or a thrombin inhibitor such as a boropeptide, hirudin or argatroban, or a thrombolytic* agent such as tissue plasminogen activator, anistreplase, urokinase or streptokinase, or combinations thereof, in one or more sterile containers, are also within the ambit of the present invention.
• an anti-coagulant agent such as warfarin or heparin
• an antiplatelet agent such as aspirin, piroxicam or ticlopidine
• kits may further include, if desired, one or more of various conventional pharmaceutical kit components, such as for example, one or more pharmaceutically acceptable carriers, additional vials for mixing the components, etc., as will be readily apparent to those skilled in the art. Instructions, either as inserts or as labels, indicating quantities of the components to be administered, guidelines for administration, and/or guidelines for mixing the components, may also be included in the kit .
• the Tables below set forth representative compounds of the present invention.
• the biological activity of the compounds is indicated as the IC 50 value in the platelet aggregation assay described above.
• uM means micromolar.

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• 1994
  • 1994-03-28 EP EP94911703A patent/EP0691987A1/en not_active Withdrawn
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