Glycosylation-mediated inhibition of factor X

• US 5,798,332 A
• Filed: 09/06/1994
• Issued: 08/25/1998
• Est. Priority Date: 03/20/1992
• Status: Expired due to Fees

• Current Assignee: Millennium Pharmaceuticals Incorporated (Takeda Pharmaceutical Company Limited)
• Original Assignee: COR Therapeutics, Inc. (Takeda Pharmaceutical Company Limited)
• Inventors: Sinha, Uma, Wolf, David L.
• Primary Examiner(s): Lankford, Jr., Leon B.
• Assistant Examiner(s): PRATS, FRANCISCO CHANDLER
• Application Number:
  US08/302,226
• Time in Patent Office: 1,449 Days
• Field of Search: 424/94.1, 424/94.5, 530/380, 530/381, 530/384, 514/8, 514/2
• US Class Current: 514/14.9
• CPC Class Codes: A61K 38/1709 from mammals
• CPC Class Codes: A61K 38/47 acting on glycosyl compounds (3.2), e.g. cellulases, lactases
• CPC Class Codes: A61P 29/00 Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
• CPC Class Codes: A61P 7/02 Antithrombotic agents; Anticoagulants; Platelet aggregation inhibitors
• CPC Class Codes: C07K 14/42 Lectins, e.g. concanavalin, phytohaemagglutinin
• CPC Class Codes: C07K 14/745 Blood coagulation or fibrinolysis factors
• CPC Class Codes: C07K 16/36 against blood coagulation factors
• CPC Class Codes: C12N 9/6432 Coagulation factor Xa (3.4.21.6)
• CPC Class Codes: C12Y 304/21006 Coagulation factor Xa (3.4.21.6)
• CPC Class Codes: Y10S 436/827 Lectins

Abstract

Reagents that specifically bind a carbohydrate target wherein sialic acid is linked at the nonreducing terminus of a glycoside to a galactose or galactosamine residue through an α2-6 linkage are able to inhibit the conversion of human Factor X to human Factor Xa. These reagents (SA/Gal/GalNAc binding reagents) as well as other strategies for inhibiting the conversion of Factor X to Factor Xa are useful in treating thrombosis, inflammation and other conditions associated with excess thrombin activity.

Claims

1. A method to treat thrombosis and conditions whose pathogenesis involves thrombin generation in a human subject, which method comprises administering to a subject in need of such treatment an amount of a reagent that binds to SA(α

2-6)Gal or SA(α

2-6)GalNAc or a pharmaceutical composition thereof effective to inhibit thrombosis.

5. The method according to any one of claims 1-4 wherein said reagent is selected from the group consisting of lectins, antibodies specifically immunoreactive to SA(α

2-6)Gal or SA(α

2-6)GalNAc, and peptides derived from Factors IXa/VIIIa or from tissue factor/Factor VIIa that specifically bind with SA(α

2-6)Gal or SA(α

2-6)GalNAc.

2. A method to treat inflammation resulting from thrombosis or conditions whose pathogenesis involves thrombin generation in a human subject, which method comprises administering to a subject in need of such treatment an amount of a reagent that binds to SA(α

2-6)Gal or SA(α

2-6)GalNAc or pharmaceutical composition thereof effective to inhibit said inflammation.

3. A method to treat restenosis resulting from thrombosis and conditions whose pathogenesis involves thrombin generation in a human subject, which method comprises administering to a subject in need of such treatment an amount of a reagent that binds to SA(α

2-6)Gal or SA(α

2-6)GalNAc or pharmaceutical composition thereof effective to inhibit said restenosis.

4. A method to treat inflammation as a complication of transplantation, said inflammation resulting from thrombosis and conditions whose pathogenesis involves thrombin generation in a human subject, which method comprises administering to a subject in need of such treatment an amount of a reagent that binds to SA(α

2-6)Gal or SA(α

2-6)GalNAc or pharmaceutical composition thereof effective to inhibit thrombotic complications of transplantation.

6. A method to inhibit the conversion of human Factor X to human Factor Xa which method comprises treating human Factor X with a reagent that binds to SA(α

2-6)Gal or SA(α

2-6)GalNAc in an amount and for a time sufficient to inhibit said conversion.

7. A method to identify a reagent that inhibits the conversion of human Factor X to human Factor Xa, which method comprises;

treating a disaccharide moiety which is SA(α

2-6)Gal or SA(α

2-6)GalNAc with a reagent;

assessing whether said reagent binds or does not bind to said disaccharide; and

determining whether any reagent that binds to said disaccharide inhibits the conversion of human Factor X to human Factor Xa.

8. The method of claim 7 wherein said reagent is a lectin.

9. The method of claim 8 wherein the lectin is a mammalian lectin.

10. A complex comprising human Factor X and a reagent identified by the method of claim 7 that binds to a disaccharide moiety which is SA(α

2-6)Gal or SA(α

2-6)GalNAc and that inhibits conversion of human Factor X to human Factor Xa.

11. A complex comprising human Factor X and a reagent that binds to SA(α

2-6)Gal or SA(α

2-6)GalNAc.

12. A complex comprising human Factor X and Sambucus nigra agglutinin (SNA).

13. A method of identifying a reagent that inhibits the conversion of human factor X to human factor Xa, which method comprises:

(a) coupling a disaccharide moiety to a solid support, wherein the disaccharide moiety is SA(α

2-6)Gal or SA(α

2-6)GalNAc present on the glycosylation side chains of human Factor X;

(b) treating the solid support with a candidate reagent which may bind to the disaccharide moiety SA(α

2-6)Gal or SA(α

2-6)GalNAc;

(c) washing the treated support to remove any unbound reagent;

(d) detecting any reagent that remains bound to the support; and

(e) further assessing whether the reagent that binds to SA(α

2-6)Gal or SA(α

2-6)GalNAc inhibits conversion of human Factor X to human Factor Xa.

14. A method to prevent thrombosis in a human subject, which method comprises administering to a subject in need of such treatment an amount of a reagent that binds to SA(α

2-6)Gal or SA(α

2-6)GalNAc or a pharmaceutical composition thereof effective to inhibit thrombosis.

18. The method according to any one of claims 14-17, wherein said reagent is selected from the group consisting of lectins, antibodies specifically immunoreactive to SA(α

2-6)Gal or SA(α

2-6)GalNAc, and peptides derived from Factors IXa/VIIIa or from tissue factor/Factor VIIa that specifically bind with SA(α

2-6)Gal or SA(α

2-6)GalNAc.

15. A method to prevent inflammation resulting from thrombosis in a human subject, which method comprises administering to a subject in need of such treatment an amount of a reagent that binds to SA(α

2-6)Gal or SA(α

2-6)GalNAc or pharmaceutical composition thereof effective to inhibit said inflammation.

16. A method to prevent restenosis resulting from thrombosis in a human subject, which method comprises administering to a subject in need of such treatment an amount of a reagent that binds to SA(α

2-6)Gal or SA(α

2-6)GalNAc or pharmaceutical composition thereof effective to inhibit said restenosis.

17. A method to prevent inflammation as a complication of transplantation, said inflammation resulting from thrombosis in a human subject, which method comprises administering to a subject in need of such treatment an amount of a reagent that binds to SA(α

2-6)Gal or SA(α

2-6)GalNAc or pharmaceutical composition thereof effective to inhibit thrombotic complications of transplantation.