Claims
- 1. A reagent for preparing a thrombus imaging agent, comprising a radiolabel complexing moiety covalently linked to a compound having a molecular weight of less than 10,000 daltons, wherein the compound specifically binds to platelet glycoprotein IIb/IIIa receptor, and wherein the reagent is capable of inhibiting human platelet aggregation in platelet-rich plasma by 50% (IC.sub.50) when present at a concentration not greater than about 1 .mu.M.
- 2. The reagent of claim 1 wherein the compound is a platelet glycoprotein IIb/IIIa receptor binding peptide having from 4 to 100 amino acids.
- 3. The reagent of claim 1 wherein the radiolabel complexing moiety has a formula selected from the group consisting of:
- Cp(aa)Cp I.
- wherein Cp is a protected cysteine and (aa) is any primary .alpha.- or .beta.-amino acid not containing a thiol group; and ##STR5## wherein X=H or a protecting group;
- (amino acid)=any primary .alpha.- or .beta.-amino acid not containing a thiol group; ##STR6## wherein each R.sup.5 is independently H, CH.sub.3 or C.sub.2 H.sub.5 ;
- each (pgp).sup.s is independently a thiol protecting group or H;
- m, n and p are independently 2 or 3;
- A=linear or cyclic lower alkyl, aryl, heterocyclyl, combinations or substituted derivatives thereof;
- X=a platelet glycoprotein IIb/IIIa receptor binding compound; and ##STR7## wherein each R.sup.5 is independently H, lower alkyl having 1 to 6 carbon atoms, phenyl, or phenyl substituted with lower alkyl or lower alkoxy;
- m, n and p are independently 1 or 2;
- A=linear or cyclic lower alkyl, aryl, heterocyclyl, combinations or substituted derivatives thereof;
- V=H or --CO-- platelet glycoprotein IIb/IIIa receptor binding compound;
- R.sup.6 =H or platelet glycoprotein IIb/IIIa receptor binding compound;
- and wherein when V=H, R.sup.6 =platelet glycoprotein IIb/IIIa receptor binding compound and when R.sup.6 =H, V=--CO-- platelet glycoprotein IIb/IIIa receptor binding compound.
- 4. The reagent of claim 1 wherein the compound and the moiety are covalently linked through one or more amino acids.
- 5. The reagent of claim 3 wherein the protected cysteine of formula I has a protecting group of the formula
- --CH.sub.2 --NH--CO--R
- wherein R is a lower alkyl having 1 to 6 carbon atoms, 2-,3-,4-pyridyl, phenyl, or phenyl substituted with lower alkyl, hydroxy, lower alkoxy, carboxy, or lower alkoxycarbonyl.
- 6. The reagent of claim 3 wherein the radiolabel complexing moiety has the formula: ##STR8##
- 7. The reagent of claim 2 wherein the peptide is selected from the group consisting of:
- CH.sub.2 CO.Y.sub.D.Apc.GDCGGG
- CH.sub.2 CO.Y.sub.D.Apc.GDCKG
- CH.sub.2 CO.Y.sub.D.Apc.GDCGG
- CH.sub.2 CO.Y.sub.D.Apc.GDC
- CH.sub.2 CO.Y.sub.D.Apc.GDCK
- CH.sub.2 CO.Y.sub.D.Amp.GDC
- CH.sub.2 CO.Y.sub.D.Amp.GDCK
- and O--(4-piperidinyl)butyl tyrosine.
- 8. A multimeric reagent for preparing a thrombus imaging agent comprising:
- a polyvalent linker covalently linked to
- a) at least two compounds that specifically bind to a platelet glycoprotein IIb/IIIa receptor; and
- b) at least one radiolabel complexing moiety;
- wherein the reagent has a molecular weight of less than about 20,000 daltons, and wherein the reagent is capable of inhibiting human platelet aggregation in platelet-rich plasma by 50 % (IC.sub.50) when present at a concentration not greater than about 1 .mu.M.
- 9. The reagent of claim 8 wherein the polyvalent linker is bis-succinimidylmethylether, 4-(2,2-dimethylacetyl)benzoic acid, N-{2-(N',N'-bis(2-succinimido-ethyl)aminoethyl)}-N.sup.6,N.sup.9 -bis(2-methyl-2-mercaptopropyl)-6,9-diazanonanamide, tris(succinimidylethyl)amine, tris(acetamidoethyl)amine, bis-(acetamidoethyl)ether, bis-(acetamidomethyl)ether, .alpha.,.epsilon.-bisacetyllysine, lysine and 1,8-bis-acetamido-3,6-dioxa-octane. 1,2-bis(2-chloroacetamidoethoxy)ethane, or a derivative thereof.
- 10. A process for preparing the reagent of claim 2 wherein the peptide is chemically synthesized in vitro.
- 11. The process of claim 10 wherein the peptide is synthesized by solid phase peptide synthesis.
- 12. The reagent of claim 2 wherein the radiolabel complexing moiety is covalently linked to the peptide during in vitro chemical synthesis.
- 13. The reagent of claim 12 wherein the radiolabel complexing moiety is covalently linked to the peptide during solid phase peptide synthesis.
- 14. The reagent of claim 1, wherein the compound comprises a cyclic peptide platelet glycoprotein IIb/IIa receptor binding domain having the formula: ##STR9## wherein A is a lipophilic D-.alpha.-amino acid, or an N-alkyl-L-.alpha.-amino acid or L-proline;
- X is an L-.alpha.-amino acid having a sidechain capable of being positively charged; and
- R is each independently H, lower alkyl or lower alkoxyalkyl.
- 15. The reagent of claim 14, wherein A is D-tyrosine or D-phenylalanine and X is L-(S-(3-aminopropyl)cysteine) or L-4-amidinophenylalanine.
- 16. The reagent of claim 1 wherein the radiolabel complexing moiety comprises a single thiol-containing moiety of formula:
- A--CZ(B)--{C(R.sup.1 R.sup.2)}.sub.n --X II.
- wherein
- A is H, HOOC, H.sub.2 NOC, (amino acid or peptide)--NHOC, (amino acid or peptide)--OOC or R.sup.4 ;
- B is H, SH, --NHR.sup.3, --N(R.sup.3)-(amino acid or peptide), or R.sup.4 ;
- X is H, SH, --NHR.sup.3, --N(R.sup.3)-(amino acid or peptide) or R.sup.4 ;
- Z is H or R.sup.4 ;
- R.sup.1, R.sup.2, R.sup.3 and R.sup.4 are independently H or lower straight or branched chain or cyclic alkyl;
- n is 0, 1 or 2;
- (peptide) is a peptide of 2 to about 10 amino acids;
- and
- where B is --NHR.sup.3 or --N(R.sup.3)-(amino acid or peptide), X is SH, and n is 1 or 2;
- where X is --NHR.sup.3 or --N(R.sup.3)-(amino acid or peptide), B is SH, and n is 1 or 2;
- where B is H or R.sup.4, A is HOOC, H.sub.2 NOC, (amino acid or peptide)--NHOC, (amino acid or peptide)--OOC, X is SH, and n is 0 or 1;
- where A is H or R.sup.4, then where B is SH, X is --NHR.sup.3 or --N(R.sup.3)-(amino acid or peptide) and where X is SH, B is --NHR.sup.3 or --N(R.sup.3)-(amino acid or peptide);
- where X is H or R.sup.4, A is HOOC, H.sub.2 NOC, (amino acid or peptide)--NHOC, (amino acid or peptide)--OOC and B is SH;
- where Z is methyl, X is methyl, A is HOOC, H.sub.2 NOC, (amino acid or peptide)--NHOC, (amino acid or peptide)--OOC, B is SH and n is 0;
- and wherein the thiol moiety is in the reduced form and (amino acid) is any primary .alpha.- or .beta.-amino acid not containing a thiol group.
- 17. The reagent of claim 16 wherein the radiolabel complexing moiety is selected from the group consisting of:
- IIa. -(amino acid).sup.1 -(amino acid).sup.2 -{A--CZ(B)--{C(R.sup.1 R.sup.2)}.sub.n --X},
- IIb. -{A--CZ(B)-{C(R.sup.1 R.sup.2)}.sub.n --X}-(amino acid).sup.1 -(amino acid).sup.2,
- IIc. -(a primary .alpha.,.omega.- or .beta.,.omega.-diamino acid)-(amino acid).sup.1 -{A--CZ(B)--{C(R.sup.1 R.sup.2)}.sub.n --X}, or
- IId. -{A--CZ(B)--{C(R.sup.1 R.sup.2)}.sub.n --X}-(amino acid).sup.1 -(a primary .alpha.,.omega.- or .beta.,.omega.-diamino acid)
- wherein
- (amino acid).sup.1 and (amino acid).sup.2 are each independently any naturally-occurring, modified, substituted or altered .alpha.- or .beta.-amino acid not containing a thiol group;
- A is H, HOOC, H.sub.2 NOC, (amino acid or peptide)--NHOC, (amino acid or peptide)--OOC or R.sup.4 ;
- B is H, SH or --NHR.sup.3, --N(R.sup.3)-(amino acid or peptide) or R.sup.4 ;
- X is SH or --NHR.sup.3, --N(R.sup.3)-(amino acid or peptide) or R.sup.4 ;
- Z is H or R.sup.4 ;
- R.sup.1, R.sup.2, R.sup.3 and R.sup.4 are independently H or straight or branched chain or cyclic lower alkyl;
- (peptide) is a peptide of 2 to about 10 amino acids;
- n is an integer that is either 0, 1 or 2; and
- where B is --NHR.sup.3 or --N(R.sup.3)-(amino acid or peptide), X is SH and n is 1 or 2;
- where X is --NHR.sup.3 or --N(R.sup.3)-(amino acid or peptide), B is SH and n is 1 or 2;
- where B is H or R.sup.4, A is HOOC, H.sub.2 NOC, (amino acid or peptide)--NHOC, (amino acid or peptide)--OOC, X is SH and n is 0 or 1;
- where A is H or R.sup.4, then where B is SH, X is --NHR.sup.3 or --N(R.sup.3)-(amino acid or peptide) and where X is SH, B is --NHR.sup.3 or --N(R.sup.3)-(amino acid or peptide);
- where X is H or R.sup.4, A is HOOC, H.sub.2 NOC, (amino acid or peptide)--NHOC, (amino acid or peptide)--OOC and B is SH;
- where Z is methyl, X is methyl, A is HOOC, H.sub.2 NOC, (amino acid or peptide)--NHOC, (peptide)--OOC and B is SH and n is 0;
- and wherein the thiol moiety is in the reduced form.
- 18. A composition of matter having the formula:
- CH.sub.2 CO--Y.sub.D ApcGDCGGC.sub.Acm GC.sub.Acm GGC.amide.
- 19. A composition of matter comprising a cyclic peptide having a molecular weight of less than 10,000 daltons wherein the peptide specifically binds to a platelet glycoprotein IIb/IIIa receptor and is capable of inhibiting human platelet aggregation in platelet-rich plasma by 50 % (IC.sub.50) when present at a concentration not greater than about 1 .mu.M, wherein the peptide comprises the sequence -Amp-Gly-Asp-.
- 20. The composition of matter of claim 19, wherein the peptide comprises the formula: ##STR10## wherein A is a lipophilic D-.alpha.-amino acid, or an N-alkyl-L-.alpha.-amino acid or L-proline; and
- R is each independently H, lower alkyl or lower alkoxyalkyl.
- 21. The composition of matter of claim 19, selected from the group consisting of:
- CH.sub.2 CO.Y.sub.D.Amp.GDC and
- CH.sub.2 CO.Y.sub.D.Amp.GDCK.
- 22. A composition of matter selected from the group consisting of cyclic peptides having the formula:
- CH.sub.2 CO.Y.sub.D.Apc.GDCGGG
- CH.sub.2 CO.Y.sub.D.Apc.GDCKG
- CH.sub.2 CO.Y.sub.D.Apc.GDCGG
- CH.sub.2 CO.Y.sub.D.Apc.GDC
- CH.sub.2 CO.Y.sub.D.Apc.GDCK
- CH.sub.2.COY.sub.D GDC
- CH.sub.2 CO.Y.sub.D.Amp.GDCK
- and O--(4-piperidinyl)butyl tyrosine.
- 23. A composition of matter having the formula:
- CH.sub.2 CO--Y.sub.D AmpGDCKGCG.amide.
Parent Case Info
This application is a continuation-in-part of International Patent Application PCT/US94/03878, filed Apr. 8, 1994, which is a continuation-in-part of U.S. patent application Ser. No. 08/044,825, filed Apr. 8, 1993 now abandoned, which is a continuation-in-part of U.S. patent application Ser. No. 07/653,012, filed Feb. 8, 1991 and now abandoned.
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Continuation in Parts (2)
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44825 |
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Parent |
653012 |
Feb 1991 |
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