Immobilised biological entities

• US 9,101,696 B2
• Filed: 03/09/2012
• Issued: 08/11/2015
• Est. Priority Date: 03/11/2011
• Status: Active Grant

• Current Assignee: W. L. Gore & Associates (W.L. Gore & Associates)
• Original Assignee: WL Gore and Associates Incorporated (W.L. Gore and Associates)
• Inventors: Leontein, Karin, Begovac, Paul, Pietrzak, Krzysztof, Antoni, Per, Nystrm, Daniel
• Primary Examiner(s): CHANNAVAJJALA, LAKSHMI SARADA
• Application Number:
  US13/416,880
• Publication Number:
  US 20120231043A1
• Time in Patent Office: 1,250 Days
• US Class Current: 1/1
• CPC Class Codes: A61L 2300/42 Anti-thrombotic agents, anticoagulants, anti-platelet agents
• CPC Class Codes: A61L 2300/452 Lubricants
• CPC Class Codes: A61L 27/34 Macromolecular materials
• CPC Class Codes: A61L 29/085 Macromolecular materials
• CPC Class Codes: A61L 31/10 Macromolecular materials
• CPC Class Codes: A61L 33/0011 Anticoagulant, e.g. heparin, platelet aggregation inhibitor, fibrinolytic agent, other than enzymes, attached to the substrate
• CPC Class Codes: A61L 33/0035 using a polymer with positively charged atoms in the polymeric backbone, e.g. ionenes
• CPC Class Codes: A61L 33/0064 Hydrogels or hydrocolloids
• CPC Class Codes: A61L 33/0076 Chemical modification of the substrate
• CPC Class Codes: A61L 33/068 obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds A61L33/062 takes precedence
• CPC Class Codes: A61L 33/08 Polysaccharides
• CPC Class Codes: A61P 7/02 Antithrombotic agents; Anticoagulants; Platelet aggregation inhibitors
• CPC Class Codes: C08G 81/00 Macromolecular compounds obtained by interreacting polymers in the absence of monomers, e.g. block polymers involving only carbon-to-carbon unsaturated bond reactions C08F299/00 ; polyester-amides C08G69/44; polyester-imides C08G73/16; polyamides-imides C08G73/14; block- or graft polymers containing polysiloxane sequences C08G77/42
• CPC Class Codes: C08L 101/005 Dendritic macromolecules
• CPC Class Codes: C08L 79/02 Polyamines
• CPC Class Codes: C08L 79/08 Polyimides; Polyester-imides; Polyamide-imides; Polyamide acids or similar polyimide precursors
• CPC Class Codes: Y10T 428/31971 Of carbohydrate

Abstract

There is described inter alia a device having a surface comprising a layered coating wherein the outer coating layer comprises a plurality of cationic hyperbranched polymer molecules characterized by having (i) a core moiety of molecular weight 14-1,000 Da (ii) a total molecular weight of 1,500 to 1,000,000 Da (iii) a ratio of total molecular weight to core moiety molecular weight of at least 80:1 and (iv) functional end groups, whereby one or more of said functional end groups have an anti-coagulant entity covalently attached thereto.

Claims

1. A device having a surface comprising a layered coating wherein the outer coating layer comprises a plurality of cationic hyperbranched polymer molecules characterized by having (i) a core moiety of molecular weight 14-1,000 Da (ii) a total molecular weight of 10,000 to 300,000 Da (iii) a ratio of total molecular weight to core moiety molecular weight of at least 80:

1 and (iv) functional end groups, whereby one or more of said functional end groups have a heparin moiety covalently attached thereto, wherein said heparin moiety is single point covalently attached to the hyperbranched polymer molecule, and wherein the hyperbranched polymer is not a dendrimer.

2. A device according to claim 1, wherein the heparin moieties are full length (native) heparin moieties.

3. A device according to claim 1, wherein the heparin moieties are nitrous acid degraded heparin moieties.

4. A device according to claim 1, wherein the heparin moieties are attached to the hyperbranched polymer molecules via the reducing end of the heparin moiety.

5. A device according to claim 1, wherein the hyperbranched polymer is selected from polyamidoamine, polypropyleneimine, polyethyleneimine and polyamine polymers and copolymers comprising one or more of polyamidoamine, polypropyleneimine, polyethyleneimine and polyamine hyperbranched polymers.

6. A device according to claim 5 wherein the hyperbranched polymer is a polyamidoamine.

7. A device according to claim 5 wherein the hyperbranched polymer is a polyethyleneimine.

8. A device according to claim 1, wherein the functional end groups are primary amine groups.

9. A device according to claim 1, wherein the core moiety has a molecular weight of 50 to 130 Da.

10. A device according to claim 9 wherein the hyperbranched polymer has ethylenediamine as core moiety.

11. A device according to claim 1, wherein the hyperbranched polymer has a molecular weight of 25,000 to 200,000 Da.

12. A device according to claim 1, wherein the ratio of total molecular weight to core moiety molecular weight is at least 100:

1.

13. A device according to claim 12 wherein the ratio of total molecular weight to core moiety molecular weight is between 200:

1 and 5000;

1 e.g. between 200;

1 and 1600;

1.

14. A device according to claim 1, wherein the hyperbranched polymer molecule has a diameter of 5 to 30 nm.

15. A device according to claim 1, wherein the hyperbranched polymer molecules are cross-linked to form aggregates of two or more hyperbranched polymer molecules.

16. A device according to claim 1, wherein the heparin moieties are covalently attached to the hyperbranched polymer molecule via a linker comprising:

i. a secondary amine linkage;

ii. an amide linkage;

iii. a thioether linkage;

or

vi. a 1,2,3-triazole linkage.

17. A device according to claim 1, wherein the heparin moieties are covalently attached to functional end groups of the hyperbranched polymer molecule via a linker formed through a free radical initiated reaction.

18. A device according to claim 16, wherein a spacer separates the linker from the hyperbranched polymer molecule and/or the heparin moiety.

19. A device according to claim 18 wherein the spacer comprises a PEG chain.

20. A device according to claim 1, wherein the layered coating comprises one or more coating bilayers of cationic polymer and anionic polymer, the innermost layer preferably being a layer of cationic polymer.

21. A device according to claim 1, wherein one or more of the layers of the layered coating other than the outer coating layer comprises a cationic hyperbranched polymer molecule characterized by having (i) a core moiety of molecular weight 14-1,000 Da (ii) a total molecular weight of 10,000 to 300,000 Da (iii) a ratio of total molecular weight to core moiety molecular weight of at least 80:

1 and (iv) functional end groups which are optionally derivatised.

22. A device according to claim 20 wherein the anionic polymer is an anionic polysaccharide.

23. A device according to claim 20 wherein the anionic polymer is a functionalized hyperbranched cationic polymer with a net negative charge.

24. A device according to claim 1, wherein hyperbranched polymer molecules of the outer coating layer are cross-linked to other hyperbranched polymer molecules of the outer coating layer or are cross-linked to molecules of an underlayer.

25. A device according to claim 1 which is a medical device.

26. A device according to claim 1 which is an analytical or separation device.

27. A device according to claim 1 wherein the layered coating of the device comprises one or more beneficial agents besides the heparin moieties.

28. A device according to claim 27 where said one or more beneficial agents are selected from drug molecules and lubricious agents.

29. A non-thrombogenic device which is obtainable by a process comprising:

(a) treating a device to present a surface coating comprising an outer coating layer comprising cationic hyperbranched polymer molecules characterized by having (i) a core moiety of molecular weight 14-1,000 Da (ii) a total molecular weight of 10,000 to 300,000 Da and (iii) a ratio of total molecular weight to core moiety molecular weight of at least 80;

1 (e.g. at least 100;

1) and bearing functional end groups, wherein the hyperbranched polymer is not a dendrimer;

(b) reacting at least one or more of said functional end groups with molecules of a heparin moiety which is functionalized to bear groups which are capable of reacting with the reactive functional groups on the hyperbranched cationic polymer;

thereby to attach the heparin moiety to the hyperbranched cationic polymer, wherein the heparin moiety is single point covalently attached to the hyperbranched polymer.

30. A non-thrombogenic device which is obtainable by a process comprising:

(a) treating a device to present a positively charged polymer surface layer; and

(b) associating with said polymer surface layer functionalized cationic hyperbranched polymer molecules characterized by having (i) a core moiety of molecular weight 14-1,000 Da (ii) a total molecular weight of 10,000 to 300,000 Da and (iii) a ratio of total molecular weight to core moiety molecular weight of at least 80;

1 (e.g. at least 100;

1) and bearing a multiplicity of negatively charged heparin moieties and wherein said functionalized hyperbranched cationic polymer has a net negative charge and is not a dendrimer;

or

(a) treating a device to present a negatively charged polymer surface layer; and

(b) associating with said polymer surface layer functionalized cationic hyperbranched polymer molecules characterized by having (i) a core moiety of molecular weight 14-1,000 Da (ii) a total molecular weight of 10,000 to 300,000 Da and (iii) a ratio of total molecular weight to core moiety molecular weight of at least 80;

1 (e.g. at least 100;

1) and bearing one or more negatively charged anti-coagulant entities such as heparin moieties and wherein said functionalized hyperbranched polymer has a net positive charge and is not a dendrimer, wherein the heparin moieties are single point covalently attached to the hyperbranched polymer.