Claims for Patent: 5,968,902

Summary for Patent: 5,968,902

Title:	Platelet aggregation inhibitors
Abstract:	The isolated and purified PAI from several active snake venoms is described and characterized. In addition, PAIs lacking the Arg-Gly-Asp (RGD) adhesion sequence but containing K* -(G/Sar)-D wherein K* is a modified lysyl residue of the formulaR12 N(CH 2)4CHNHCO-wherein each R1 is independently H, alkyl(1-6C) or at most one R1 is R2-C=NR3 wherein R2 is H, alkyl(1-6C), phenyl or benzyl, or is NR4 2 in which each R4 is independently H or alkyl(1-6C) and R3 is H, alkyl(1-6C), phenyl or benzyl, or R2-C=NR3 is a radical selected from the group consisting of: where m is an integer of 2-3, and each R5 is independently H or alkyl(1-6C); and wherein one or two (CH2) may be replaced by O or S provided said O or S is not adjacent to another heteroatom are prepared and shown to specifically inhibit the binding of fibrinogen or von Willebrand Factor to GP IIb-IIIa.
Inventor(s):	Robert M. Scarborough, David Lawrence Wolf, Israel F. Charo
Assignee:	COR Therapeutics Inc , Millennium Pharmaceuticals Inc
Application Number:	US08/482,263
Patent Claims:	1. A method of treating or preventing a platelet associated ischemic disorder in a patient comprising administering to said patient an effective amount of a platelet aggregation inhibitor of the formula: ##STR11## (1) where K* is a lysyl residue of the formula R.sup.1.sub.2 N(CH.sub.2).sub.4 CHNHCO-- wherein each R1 is independently hydrogen, alkyl(1-6C) or one R1 is R2 --C═NR3, in which:R2 is hydrogen, alkyl(1-6C), a substituted or unsubstituted phenyl or benzyl residue, or NR4 2 in which each R4 is independently hydrogen or alkyl(1-6C), and R3 is hydrogen, alkyl(1-6C), phenyl or benzyl, or R2 --C═NR3 is a radical selected from the group consisting of ##STR12## wherein m is an integer of 2-3, and each R5 is independently H or alkyl(1-6C) and one or two --CH2 -- may be replaced by O or S provided said O or S is not adjacent to another heteroatom; (2) where AA1 and AA4 are each independently selected from the group consisting of Gly, Ala, and Ser; n1 is an integer of 0-3; and n4 is an integer of 0-3; (3) where AA2 is selected from the group consisting of tryptophan, phenylalanine, leucine, tyrosine, and valine and n2 is an integer of 0-3; (4) where AA3 is a proline residue or a modified proline residue of the formula ##STR13## wherein one or two of the methylenes of said proline or modified proline residue is optionally replaced by --NR--, --S--, or --O-- wherein R is hydrogen or alkyl (1-6C) and n3 is an integer of 0-1; (5) where each of X1 and X2 is independently a residue selected from the group consisting of cysteine, mercaptopropionyl, mercaptovaleryl, and penicillamine, and ##STR14## represents a bond between X1 and X2 ; and (6) where each of Y1 and Y2 is independently a non-interfering substituent or is absent; and (7) where one or more peptide linkages may optionally be replaced by a linkage selected from the group consisting of --CH2 NH--, --CH2 S--, --CH2 CH2 --, --CH═CH-- (cis or trans), --COCH2 --, --CH(OH)CH2 -- and --CH2 SO--; and (8) where all chiral amino acid residues in said formula are of the L configuration; (9) with the proviso that if n3 is 0; either:1) the sum of n2 and n4 must be at least 2; or 2) K* cannot be Har or Lys; or 3) X2 cannot be cysteine, penicillamine, or 2-amino-3,3-cyclopentanemethylene-3-mercaptopropionic acid; or 4) one or more peptide linkages is replaced by said alternate linkage, or a physiologically acceptable basic or acid addition salt thereof. 2. A method according to claim 1, wherein said platelet aggregation inhibitor is selected from the group consisting of:Gly-Cys-Gly-Lys-Gly-Asp-Trp-Pro-Cys-Ala-NH2 ; Gly-Cys-Lys-Gly-Asp-Trp-Pro-Cys-Ala-NH2 ; Cys-Gly-Lys-Gly-Asp-Trp-Pro-Cys-NH2 ; Cys-Lys-Gly-Asp-Trp-Pro-Cys-NH2 ; Cys-Lys-Gly-Asp-Tyr-Pro-Cys-NH2 ; Cys-Lys-Gly-Asp-Phe-Pro-Cys-NH2 ; Cys-Lys-Gly-Asp-Leu-Pro-Cys-NH2 ; Cys-Lys-Gly-Asp-Val-Pro-Cys-NH2 ; Mpr-Lys-Gly-Asp-Trp-Pro-Cys-NH2 ; Mpr-Lys-Gly-Asp-Tyr-Pro-Cys-NH2 ; Mpr-Lys-Gly-Asp-Phe-Pro-Cys-NH2 ; Mpr-Lys-Gly-Asp-Leu-Pro-Cys-NH2 ; Mpr-Lys-Gly-Asp-Val-Pro-Cys-NH2 ; Cys-Lys-Gly-Asp-Trp-Gly-Cys-NH2 ; Cys-Lys-Sar-Asp-Trp-Pro-Cys-NH2 ; Acetyl-Cys-Lys-Gly-Asp-Trp-Pro-Cys-NH2 ; Mvl-Lys-Gly-Asp-Trp-Pro-Cys-NH2 ; Mpr-Lys-Gly-Asp-Trp-Pro-Pen-NH2 ; Mpr-Lys-Gly-Asp-Trp-Thz-Cys-NH2 ; Mvl-Lys-Gly-Asp-Trp-Pro-Pen-NH2 ; Mpr-Lys-Gly-Asp-Trp-Pip-Pen-NH2 ; Mpr-Har-Gly-Asp-Trp-Pro-Cys-NH2 ; Mpr-Har-Gly-Asp-Trp-Pro-Pen-NH2 ; Mpr-(Acetimidyl-Lys)-Gly-Asp-Trp-Pro-Cys-NH2 ; Mpr-(Acetimidyl-Lys)-Gly-Asp-Trp-Pro-Pen-NH2 ; Mpr (NG, NG' -ethylene-Har)-Gly-Asp-Trp-Pro-Cys-NH2 ; Mpr (NG, NG' -ethylene-Har)-Gly-Asp-Trp-Pro-Pen-NH2 ; Mpr-Har-Sar-Asp-Trp-Pro-Cys-NH2 ; Mpr-(Acetimidyl-Lys)-Gly-Asp-Trp-Pro-Pen-NH2 ; Mpr-(Phenylimidyl-Lys)-Gly-Asp-Trp-Pro-Cys-NH2 ; Mpr-Har-Sar-Asp-Trp-Pro-Pen-NH2 ; Mpr-(Phenylimidyl-Lys)-Gly-Asp-Trp-Pro-Pen-NH2 ; and Mpr-(Phenylimidyl-Lys)-Gly-Asp-Pen-NH2. 3. A method according to claim 1, wherein said disorder is thrombus formation. 4. A method according to claim 1, wherein said disorder is acute myocardial infarction. 5. A method according to claim 1, wherein said disorder is thrombosis following angioplasty. 6. A method according to claim 1, wherein said disorder is unstable angina. 7. A method according to claim 1, wherein said disorder is atherosclerosis. 8. A method according to claim 1, wherein said disorder is characterized by transient ischemic attacks. 9. A method according to claim 1, wherein said disorder is peripheral vascular disease. 10. A method according to claim 1, wherein said disorder is restenosis following angioplasty. 11. A method according to claim 1, wherein said disorder is thrombosis following carotid endarterectomy. 12. A method according to claim 1, wherein said disorder is thrombosis following anastomosis of vascular grafts. 13. A method of preventing platelet loss during extracorporeal circulation of blood comprising contacting said blood with an effective amount of a platelet aggregation inhibitor of the formula: ##STR15## (1) where K* is a lysyl residue of the formula R.sup.1.sub.2 N(CH.sub.2).sub.4 CHNHCO-- wherein each R1 is independently hydrogen, alkyl(1-6C) or one R1 is R2 --C═NR3, in which:R2 is hydrogen, alkyl(1-6C), a substituted or unsubstituted phenyl or benzyl residue, or NR4 2 in which each R4 is independently hydrogen or allyl(1-6C), and R3 is hydrogen, alkyl(1-6C), phenyl or benzyl, or R2 --C═NR3 is a radical selected from the group consisting of ##STR16## wherein m is an integer of 2-3, and each R5 is independently H or alkyl(1-6C) and one or two --CH2 -- may be replaced by O or S provided said O or S is not adjacent to another heteroatom; (2) where AA1 and AA4 are each independently selected from the group consisting of Gly, Ala, and Ser; n1 is an integer of 0-3; and n4 is an integer of 0-3; (3) where AA2 is selected from the group consisting of tryptophan, phenylalanine, leucine, tyrosine, and valine and n2 is an integer of 0-3; (4) where AA3 is a proline residue or a modified proline residue of the formula ##STR17## wherein one or two of the methylenes of said proline or modified proline residue is optionally replaced by --NR--, --S--, or --O-- wherein R is hydrogen or alkyl (1-6C) and n3 is an integer of 0-1; (5) where each of X1 and X2 is independently a residue selected from the group consisting of cysteine, mercaptopropionyl, mercaptovaleryl, and penicillamine, and ##STR18## represents a bond between X1 and X2 ; and (6) where each of Y1 and Y2 is independently a non-interfering substituent or is absent; and (7) where one or more peptide linkages may optionally be replaced by a linkage selected from the group consisting of --CH2 NH--, --CH2 S--, --CH2 CH2 --, --CH═CH-- (cis or trans), --COCH2 --, -CH(OH)CH2 -- and --CH2 SO--; and (8) where all chiral amino acid residues in said formula are of the L configuration; (9) with the proviso that if n3 is 0; either:1) the sum of n2 and n4 must be at least 2; or 2) K* cannot be Har or Lys; or 3) X2 cannot be cysteine, penicillamine, or 2-amino-3,3-cyclopentanemethylene-3-mercaptopropionic acid; or 4) one or more peptide linkages is replaced by said alternate linkage, or a physiologically acceptable basic or acid addition salt thereof. 14. A method according to claim 13, wherein said platelet aggregation inhibitor is selected from the group consisting of:Gly-Cys-Gly-Lys-Gly-Asp-Trp-Pro-Cys-Ala-NH2 ; Gly-Cys-Lys-Gly-Asp-Trp-Pro-Cys-Ala-NH2 ; Cys-Gly-Lys-Gly-Asp-Trp-Pro-Cys-NH2 ; Cys-Lys-Gly-Asp-Trp-Pro-Cys-NH2 ; Cys-Lys-Gly-Asp-Tyr-Pro-Cys-NH2 ; Cys-Lys-Gly-Asp-Phe-Pro-Cys-NH2, Cys-Lys-Gly-Asp-Leu-Pro-Cys-NH2 ; Cys-Lys-Gly-Asp-Val-Pro-Cys-NH2 ; Mpr-Lys-Gly-Asp-Trp-Pro-Cys-NH2 ; Mpr-Lys-Gly-Asp-Tyr-Pro-Cys-NH2 ; Mpr-Lys-Gly-Asp-Phe-Pro-Cys-NH2 ; Mpr-Lys-Gly-Asp-Leu-Pro-Cys-NH2 ; Mpr-Lys-Gly-Asp-Val-Pro-Cys-NH2 ; Cys-Lys-Gly-Asp-Trp-Gly-Cys-NH2 ; Cys-Lys-Sar-Asp-Trp-Pro-Cys-NH2 ; Acetyl-Cys-Lys-Gly-Asp-Trp-Pro-Cys-NH2 ; Mvl-Lys-Gly-Asp-Trp-Pro-Cys-NH2 ; Mpr-Lys-Gly-Asp-Trp-Pro-Pen-NH2 ; Mpr-Lys-Gly-Asp-Trp-Thz-Cys-NH2 ; Mvl-Lys-Gly-Asp-Trp-Pro-Pen-NH2 ; Mpr-Lys-Gly-Asp-Trp-Pip-Pen-NH2 ; Mpr-Har-Gly-Asp-Trp-Pro-Cys-NH2 ; Mpr-Har-Gly-Asp-Trp-Pro-Pen-NH2 ; Mpr-(Acetimidyl-Lys)-Gly-Asp-Trp-Pro-Cys-NH2 ; Mpr-(Acetimidyl-Lys)-Gly-Asp-Trp-Pro-Pen-NH2 ; Mpr (NG, NG' -ethylene-Har)-Gly-Asp-Trp-Pro-Cys-NH2 ; Mpr (NG, NG' -ethylene-Har)-Gly-Asp-Trp-Pro-Pen-NH2 ; Mpr-Har-Sar-Asp-Trp-Pro-Cys-NH2 ; Mpr-(Acetimidyl-Lys)-Gly-Asp-Trp-Pro-Pen-NH2 ; Mpr-(Phenylimidyl-Lys)-Gly-Asp-Trp-Pro-Cys-NH2 ; Mpr-Har-Sar-Asp-Trp-Pro-Pen-NH2 ; Mpr-(Phenylimidyl-Lys)-Gly-Asp-Trp-Pro-Pen-NH2 ; and Mpr-(Phenylimidyl-Lys)-Gly-Asp-Pen-NH2. 15. A method of preventing platelet aggregation, embolization or consumption of extracorporeal circulation comprising administering an effective amount of a platelet aggregation inhibitor of the formula: ##STR19## (1) where K* is a lysyl residue of the formula R.sup.1.sub.2 N(CH.sub.2).sub.4 CHNHCO-- wherein each R1 is independently hydrogen, alkyl(1-6C) or one R1 is R2 --C═NR3, in which:R2 is hydrogen, alkyl(1-6C), a substituted or unsubstituted phenyl or benzyl residue, or NR4 2 in which each R4 is independently hydrogen or alkyl(1-6C), and R3 is hydrogen, allyl(1-6C), phenyl or benzyl, or R2 --C═NR3 is a radical selected from the group consisting of ##STR20## wherein m is an integer of 2-3, and each R5 is independently H or alkyl(1-6C) and one or two --CH2 -- may be replaced by O or S provided said O or S is not adjacent to another heteroatom; (2) where AA1 and AA4 are each independently selected from the group consisting of Gly, Ala, and Ser; n1 is an integer of 0-3; and n4 is an integer of 0-3; (3) where AA2 is selected from the group consisting of tryptophan, phenylalanine, leucine, tyrosine, and valine and n2 is an integer of 0-3; (4) where AA3 is a proline residue or a modified proline residue of the formula ##STR21## wherein one or two of the methylenes of said proline or modified proline residue is optionally replaced by --NR--, --S--, or --O-- wherein R is hydrogen or alkyl (1-6C) and n3 is an integer of 0-1; (5) where each of X1 and X2 is independently a residue selected from the group consisting of cysteine, mercaptopropionyl, mercaptovaleryl, and penicillamine, and ##STR22## represents a bond between X1 and X2 ; and (6) where each of Y1 and Y2 is independently a non-interfering substituent or is absent; and (7) where one or more peptide linkages may optionally be replaced by a linkage selected from the group consisting of --CH2 NH--, --CH2 S--, --CH2 CH2 --, --CH═CH-- (cis or trans), --COCH2 --, --CH(OH)CH2 -- and --CH2 SO--; and (8) where all chiral amino acid residues in said formula are of the L configuration; (9) with the proviso that if n3 is 0; either:1) the sum of n2 and n4 must be at least 2; or 2) K* cannot be Har or Lys; or 3) X2 cannot be cysteine, penicillamine, or 2-amino-3,3-cyclopentanemethylene-3-mercaptopropionic acid; or 4) one or more peptide linkages is replaced by said alternate linkage, or a physiologically acceptable basic or acid addition salt thereof. 16. A method according to claim 15, wherein said platelet aggregation inhibitor is selected from the group consisting of:Gly-Cys-Gly-Lys-Gly-Asp-Trp-Pro-Cys-Ala-NH2 ; Gly-Cys-Lys-Gly-Asp-Trp-Pro-Cys-Ala-NH2 ; Cys-Gly-Lys-Gly-Asp-Trp-Pro-Cys-NH2 ; Cys-Lys-Gly-Asp-Trp-Pro-Cys-NH2 ; Cys-Lys-Gly-Asp-Tyr-Pro-Cys-NH2 ; Cys-Lys-Gly-Asp-Phe-Pro-Cys-NH2 ; Cys-Lys-Gly-Asp-Leu-Pro-Cys-NH2 ; Cys-Lys-Gly-Asp-Val-Pro-Cys-NH2 ; Mpr-Lys-Gly-Asp-Trp-Pro-Cys-NH2 ; Mpr-Lys-Gly-Asp-Tyr-Pro-Cys-NH2 ; Mpr-Lys-Gly-Asp-Phe-Pro-Cys-NH2 ; Mpr-Lys-Gly-Asp-Leu-Pro-Cys-NH2 ; Mpr-Lys-Gly-Asp-Val-Pro-Cys-NH2 ; Cys-Lys-Gly-Asp-Trp-Gly-Cys-NH2 ; Cys-Lys-Sar-Asp-Trp-Pro-Cys-NH2 ; Acetyl-Cys-Lys-Gly-Asp-Trp-Pro-Cys-NH2 ; Mvl-Lys-Gly-Asp-Trp-Pro-Cys-NH2 ; Mpr-Lys-Gly-Asp-Trp-Pro-Pen-NH2 ; Mpr-Lys-Gly-Asp-Trp-Thz-Cys-NH2 ; Mvl-Lys-Gly-Asp-Trp-Pro-Pen-NH2 ; Mpr-Lys-Gly-Asp-Trp-Pip-Pen-NH2 ; Mpr-Har-Gly-Asp-Trp-Pro-Cys-NH2 ; Mpr-Har-Gly-Asp-Trp-Pro-Pen-NH2 ; Mpr-(Acetimidyl-Lys)-Gly-Asp-Trp-Pro-Cys-NH2 ; Mpr-(Acetimidyl-Lys)-Gly-Asp-Trp-Pro-Pen-NH2 ; Mpr (NG, NG' -ethylene-Har)-Gly-Asp-Trp-Pro-Cys-NH2 ; Mpr (NG, NG' -ethylene-Har)-Gly-Asp-Trp-Pro-Pen-NH2 ; Mpr-Har-Sar-Asp-Trp-Pro-Cys-NH2 ; Mpr-(Acetimidyl-Lys)-Gly-Asp-Trp-Pro-Pen-NH2 ; Mpr-(Phenylimidyl-Lys)-Gly-Asp-Trp-Pro-Cys-NH2 ; Mpr-Har-Sar-Asp-Trp-Pro-Pen-NH2 ; Mpr-(Phenylimidyl-Lys)-Gly-Asp-Trp-Pro-Pen-NH2 ; and Mpr-(Phenylniidyl-Lys)-Gly-Asp-Pen-NH2. 17. A method according to claim 15, wherein said platelet aggregation, embolization or consumption is due to extracorporeal circulation for renal dialysis. 18. A method according to claim 15, wherein said platelet aggregation, embolization or consumption is due to extracorporeal circulation for cardiopulmonary bypass. 19. A method according to claim 15, wherein said platelet aggregation, embolization or consumption is due to extracorporeal circulation for hemoperfision. 20. A method according to claim 15, wherein said platelet aggregation, embolization or consumption is due to extracorporeal circulation for plasmapheresis. 21. A method according to claim 15, wherein said platelet aggregation, embolization or consumption is associated with an intravascular device. 22. A method according to claim 21, wherein said intravascular device is an intraaortic balloon pump. 23. A method according to claim 21, wherein said intravascular device is a ventricular assist device. 24. A method according to claim 21, wherein said intravascular device is an arterial catheter.