METHODS FOR REMOVING CYTOKINES FROM BLOOD WITH SURFACE IMMOBILIZED POLYSACCHARIDES

• US 20140231357A1
• Filed: 04/24/2014
• Published: 08/21/2014
• Est. Priority Date: 12/01/2009
• Status: Active Grant

• Current Assignee: ExThera Medical Corporation
• Original Assignee: ExThera Medical Corporation
• Inventors: Ward, Robert S, McCrea, Keith R., Larm, Olle, Adolfsson, Lars
• Granted Patent: US 9,408,962 B2
• Time in Patent Office: Days
• US Class Current: 210/690
• CPC Class Codes: A61K 31/60 Salicylic acid; Derivatives thereof
• CPC Class Codes: A61K 31/722 Chitin, chitosan
• CPC Class Codes: A61K 31/727 Heparin; Heparan
• CPC Class Codes: A61K 31/728 Hyaluronic acid
• CPC Class Codes: A61M 1/3472 with treatment of the filtrate
• CPC Class Codes: A61M 1/3486 Biological, chemical treatment, e.g. chemical precipitation; treatment by absorbents
• CPC Class Codes: A61M 1/3496 Plasmapheresis; Leucopheresis; Lymphopheresis A61M1/3472 takes precedence
• CPC Class Codes: A61M 1/36 Other treatment of blood in a by-pass of the natural circulatory system, e.g. temperature adaptation, irradiation ; Extra-corporeal blood circuits
• CPC Class Codes: A61M 1/362 changing physical properties of target cells by binding them to added particles to facilitate their subsequent separation from other cells, e.g. immunoaffinity
• CPC Class Codes: A61M 1/3621 Extra-corporeal blood circuits single-needle circuits A61M1/30
• CPC Class Codes: A61M 1/3673 Anticoagulant coating, e.g. Heparin coating
• CPC Class Codes: A61M 1/3679 by absorption A61M1/3675 takes precedence
• CPC Class Codes: A61M 1/3687 Chemical treatment A61M1/3675 takes precedence
• CPC Class Codes: A61M 1/38 Removing constituents from donor blood and storing or returning remainder to body , e.g. for transfusion
• CPC Class Codes: A61M 2202/0057 retained by adsorption
• CPC Class Codes: A61P 1/04 for ulcers, gastritis or reflux esophagitis, e.g. antacids, inhibitors of acid secretion, mucosal protectants
• CPC Class Codes: A61P 17/02 for treating wounds, ulcers, burns, scars, keloids, or the like
• CPC Class Codes: A61P 31/00 Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
• CPC Class Codes: A61P 31/04 Antibacterial agents
• CPC Class Codes: A61P 37/06 Immunosuppressants, e.g. drugs for graft rejection

Abstract

The present invention is directed to a method for removing cytokines and/or pathogens from blood or blood serum (blood) by contacting the blood with a solid, essentially non microporous substrate which has been surface treated with heparin, heparan sulfate and/or other molecules or chemical groups (the adsorbent media or media) having a binding affinity for the cytokine or pathogen(s) to be removed (the adsorbates), and wherein the size of the interstitial channels within said media is balanced with the amount of media surface area and the surface concentration of binding sites on the media in order to provide adequate adsorptive capacity while also allowing relatively high flow rates of blood through the adsorbent media.

Claims

1. A method for removing at least one adsorbate selected from cytokines, pathogens and toxins from blood, the method comprising:

providing a device which comprises a container, said container comprising a solid substrate of high surface area having at least one polysaccharide adsorbent on the surface thereof with a binding affinity for the adsorbate, wherein said substrate is sufficiently solid such that the adsorbate does not pass through pores in said substrate, and wherein the size of interstitial channel spaces within said substrate is large enough to permit the transport of blood cells, and the amount of interstitial surface area within said substrate is such that when flowing blood is in contact with said substrate through said device, said cytokine, pathogen or toxin binds to binding sites on said at least one polysaccharide adsorbent, becoming separated from said blood and the transport of said adsorbates from blood to adsorbent sites on said substrate is primarily by convection transport, such that adsorbate binding does not require the Brownian diffusion of adsorbates into pores within said substrate, wherein blood is flowed through the device to remove at least one adsorbate.

2. The method according to claim 1, wherein said solid substrate comprises a plurality of rigid polymer beads.

3. The method according to claim 2, wherein said rigid polymer beads are rigid polyethylene beads.

4. The method according to claim 1, wherein at least one of said polysaccharides is a member selected from the group consisting of heparin, heparan sulfate, hyaluronic acid, sialic acid, carbohydrates with mannose sequences, and chitosan.

5. The method of claim 4, wherein the linear flow velocity of blood flowing through said device is between about 8 and 24 cm/minute.

6. The method of claim 3, wherein said beads have a diameter ranging from 100 and 450 microns.

7. The method of claim 6, wherein said beads have an average diameter of 0.3 mm.

8. The method according to claim 2, wherein said beads are coated with 0.5-10 mg heparin per gram of bead.

9. The method of claim 8, wherein said beads are coated with 2±

0.5 mg heparin per gram of bead.

10. The method according to claim 9, wherein the heparin has a mean molecular weight of about 8 kDa.

11. The method according to claim 8, wherein the heparin is attached to the substrate by covalent end-point attachment.

12. The method according to claim 11, wherein said container further comprises a solid substrate having on the surface thereof a cationic, positively charged molecule having an affinity for the cytokine, pathogen or toxin, but being more thrombogenic than heparin.

13. The method of claim 12, wherein said cationic, positively charged molecule is chitosan or polyethylene imine.

14. A method for removing at least one adsorbate selected from cytokines, pathogens and toxins from blood, the method comprising:

providing a device which comprises a container that comprises a solid substrate comprised of a plurality of rigid polyethylene beads comprising heparin on the surface thereof with a binding affinity for the adsorbate, wherein said beads are sufficiently solid such that the adsorbate does not pass through pores in said beads, and wherein the size of interstitial channel spaces within said substrate is large enough to permit the transport of blood cells, and the amount of interstitial surface area within said substrate is such that when flowing blood is in contact with said substrate through said device, said cytokine, pathogen or toxin binds to binding sites on said heparin, becoming separated from said blood and the transport of said adsorbates from blood to adsorbent sites on said beads is primarily by convection transport, such that adsorbate binding does not require the Brownian diffusion of adsorbates into pores within said substrate and wherein said heparin is attached to the substrate by covalent end-point attachment in an amount of 0.5-10 mg heparin per gram of bead, wherein blood is flowed through the device to remove at least one adsorbate.

15. The method according to claim 14, wherein the linear flow velocity of blood flowing through said device is between about 8 and 24 cm/minute.

16. The method according to claim 14, wherein said beads have a diameter ranging from 100 and 450 microns.

17. The method according to claim 16, wherein said beads have an average diameter of 0.3 mm.

18. The method according to claim 14, wherein said beads are coated with 2±

0.5 mg heparin per gram of bead.

19. The method according to claim 18, wherein the heparin has a mean molecular weight of about 8 kDa.

20. The method according to claim 14, wherein said container further comprises a solid substrate having on the surface thereof a cationic, positively charged molecule having an affinity for the cytokine, pathogen or toxin, but being more thrombogenic than heparin.

21. The method according to claim 20, wherein said cationic, positively charged molecule is chitosan or polyethylene imine.

22. The method according to claim 14, wherein the cytokine is selected from the group consisting of IL-6, VCAM, TNF-α

, GRO-α

, IL-8, and RANTES.

23. A method for removing at least one adsorbate selected from cytokines, pathogens and toxins from blood, the method comprising:

providing a device which comprises a container, said container comprising a solid substrate of high surface area comprising;

i) a first portion of the solid substrate with at least one polysaccharide adsorbent on the surface, wherein said at least one polysaccharide comprises binding sites for the at least one adsorbate; and

ii) a second portion of the solid substrate having on the surface thereof a cationic, positively charged molecule being more thrombogenic than heparin, wherein said substrate is sufficiently solid such that the adsorbate does not pass through pores in said substrate, and wherein the size of interstitial channel spaces within said substrate is large enough to permit the transport of blood cells, and the amount of interstitial surface area within said substrate is such that when flowing blood is in contact with said substrate through said device, said cytokine, pathogen and toxin becoming separated from said blood and the transport of said adsorbates from blood to adsorbent sites on said substrate is primarily by convection transport, such that adsorbate binding does not require the Brownian diffusion of adsorbates into pores within said substrate, wherein blood is flowed through the device to remove at least one adsorbate.

24. The method according to claim 23, wherein the at least one of said polysaccharides is a member selected from the group consisting of heparin, heparan sulfate, hyaluronic acid, sialic acid, carbohydrates with mannose sequences, and chitosan.

25. The method according to claim 23, wherein the linear flow velocity of blood flowing through said device is between about 8 and 24 cm/minute.

26. The method according to claim 23, wherein said beads have a diameter ranging from 100 and 450 microns.

27. The method according to claim 26, wherein said beads have an average diameter of 0.3 mm.

28. The method according to claim 23, wherein said beads are coated with 2±

0.5 mg heparin per gram of bead.

29. The method according to claim 24, wherein the heparin has a mean molecular weight of about 8 kDa.

30. The method according to claim 23, wherein said cationic, positively charged molecule is chitosan or polyethylene imine.

31. The method according to claim 23, wherein the cytokine is selected from the group consisting of IL-6, VCAM, TNF-α

, GRO-α

, IL-8, and RANTES.