High affinity Siglec ligands

• US 8,357,671 B2
• Filed: 11/10/2006
• Issued: 01/22/2013
• Est. Priority Date: 11/10/2005
• Status: Active Grant

• Current Assignee: James Paulson
• Original Assignee: James Paulson
• Inventors: Paulson, James, Collins, Brian, Han, Shoufa
• Primary Examiner(s): McIntosh, III, Traviss C
• Application Number:
  US12/084,723
• Publication Number:
  US 20090238837A1
• Time in Patent Office: 2,265 Days
• Field of Search: None
• US Class Current: 514/53
• CPC Class Codes: A61K 31/655 Azo (—N=N—), diazo (=N2), azoxy (>N—O—N< or N(=O)—N<), azido (—N3) or diazoamino (—N=N—N<) compounds
• CPC Class Codes: A61K 31/715 Polysaccharides, i.e. having more than five saccharide radicals attached to each other by glycosidic linkages; Derivatives thereof, e.g. ethers, esters
• CPC Class Codes: A61K 31/739 Lipopolysaccharides
• CPC Class Codes: A61K 38/168 from plants
• CPC Class Codes: A61K 47/549 Sugars, nucleosides, nucleotides or nucleic acids
• CPC Class Codes: A61K 47/6911 the form being a liposome
• CPC Class Codes: A61P 35/00 Antineoplastic agents

Abstract

The invention relates to high affinity Siglec ligands that are useful for isolating cells that express Siglecs and for delivering agents to cells that express Siglecs. In one embodiment, the invention provides a method for treating cancer in a mammal that involves administering a Siglec ligand of the invention to the mammal, where the Siglec ligand is linked to a therapeutic agent.

Claims

1. A compound of formula Ia, Ib, Ic or Id:

[R₉-Sia-Sac-Y]_n-Carrier 

 

Ia

[R₉-Sia-Sac-Y]_n˜

Carrier 

 

Ib

[R₅-Sia-Sac-Y]_n-Carrier 

 

Ic

[R₅-Sia-Sac-Y]_n˜

Carrier 

 

Id

wherein;

R₅ and R₉ are independently each a hydrogen, hydroxyl, acetylamine, alkyl, alkylamide, cycloalkyl, heteroalkyl, cycloheteroalkyl, aryl, arylalkyl, arylamide or heteroaryl group that can be substituted with one to five alkyl, halo, sulfate or hydroxy groups and R5 and R9 are attached to different positions on the Sia;

Sia is a sialic acid;

Sac is a neutral saccharide;

Y is a covalent bond to Carrier, a linker that can be attached to Carrier or an antigen that can bind to an antibody Carrier;

the symbol ˜

is a non-covalent bond;

n is an integer of 5 or more; and

Carrier is a polymer, protein, antibody, antigen, multi-subunit protein, protein complex, glycan, lipid, liposome or solid support; and

wherein at least one of R₅ or R₉ is an alkyl, alkylamide, cycloalkyl, heteroalkyl, cycloheteroalkyl, aryl, arylalkyl, arylamide or heteroaryl group that can be substituted with one to five alkyl, halo, or sulfate groups.

2. The compound of claim 1, wherein R₅ or R₉ is an aryl, arylalkyl, arylamide or heteroaryl group.

3. The compound of claim 2, wherein the aryl, arylalkyl, arylamide or heteroaryl group comprises a phenyl ring, a bicyclic ring, or tricyclic ring, with about five to about fourteen ring atoms in which at least one ring is aromatic.

4. The compound of claim 2, wherein the aryl, arylalkyl, arylamide or heteroaryl group comprises a phenyl, biphenyl, naphthalene, anthracene, or a combination thereof.

5. The compound of claim 1, wherein the sialic acid is N-acetylneuraminic acid (Neu5Ac) or N-glycolylneuraminic acid (Neu5Gc).

6. The compound of claim 1, wherein the carrier is a polymer.

7. The compound of claim 1, wherein Y is an antigen.

8. The compound of claim 7, wherein the antigen comprises Galα

1-3Gal or nitrophenyl.

9. The compound of claim 1, wherein the carrier is an antibody.

10. The compound of claim 9, wherein the antibody is an IgM antibody.

11. The compound of claim 1, wherein the carrier is polyacrylamide, ROMP, TRIS, amyloid serum protein, glycoprotein or liposome.

12. The compound of claim 1, wherein at least one compound of formula Ia or Ic is covalently attached to the carrier.

13. The compound of claim 1, wherein at least one compound of formula Ib or Id binds to the carrier through non-covalent interactions.

22. A multivalent complex consisting of a Carrier to which 5 to 500 compounds are attached, each compound consisting of a compound of claim 1.

23. An article comprising a solid support and a compound of claim 1.

24. The article of claim 23, wherein the solid support is a magnetic bead, a chromatographic matrix, a microtiter dish or a substrate for an array of ligands.

25. A method of isolating a cell that expresses a Siglec comprising:

(a) contacting a mixture of cells that may express a Siglec with the article of claim 23;

(b) washing the article; and

(c) eluting cells from the article.

26. The method of claim 25, wherein the cell is a B cell.

27. The method of claim 25, wherein the cell is a leukemia cell or a lymphoma cell.

28. The method of claim 25, wherein the cell is an eosinophil.

29. The method of claim 25, wherein the mixture of cells is a test sample.

30. The method of claim 29, wherein the test sample is a blood sample from a patient suspected of having a B cell disorder.

31. The method of claim 29, wherein the test sample is a pulmonary exudate, lymph sample or blood sample from a patient suspected of having an eosinophil-related disease or condition.

32. A composition comprising a pharmaceutically acceptable carrier and a therapeutically effective amount of a compound of claim 1 or the multivalent complex of claim 22.

33. The composition of claim 32, wherein the compound or complex is covalently or non-covalently linked to a therapeutic agent.

34. The composition of claim 33, wherein the therapeutic agent is an anti-tumor agent, an antigen or an antibody.

35. A method for intracellular delivery of an agent into a cell, comprising contacting the cell with a compound of claim 1 or the multivalent complex of claim 22, wherein the cell expresses a Siglec and the compound is directly or indirectly linked to the agent.

36. The method of claim 35, wherein the Siglec is CD22, Siglec-8, CD33 or myelin associated glycoprotein.

37. The method of claim 35, wherein the cell is a B cell.

38. The method of claim 35, wherein the cell is a leukemia cell or a lymphoma cell.

39. The method of claim 35, wherein the cell is an eosinophil.

40. The method of claim 35, wherein the agent is an anti-tumor agent.

41. The method of claim 35, wherein the agent is an antibody.

42. The method of claim 35, wherein the agent is linked to the compound by a liposome carrier in which the agent is encapsulated and to which the compound is attached.

43. The method of claim 35 wherein the cell expresses CD22 and the Siglec ligand is a compound selected from the group consisting of:

46. A method of treating cancer in a mammal comprising administering to the mammal an effective amount of a compound of claim 1, or the multivalent complex of claim 22, wherein the compound is linked to an anti-tumor agent.

47. The method of claim 46, wherein the cancer is a lymphoma or leukemia.

14. A compound of formula IIa or IIb:

15. The compound of claim 14, wherein the R₂ and R₆ substituents are joined by a linker.

16. The compound of claim 14, wherein R₆ is a non-covalently associated carrier.

17. The compound of claim 14, wherein the aryl is a phenyl ring, a bicyclic ring, or tricyclic ring that has about five to about fourteen ring atoms and where the aryl has at least one aromatic ring.

18. The compound of claim 14, wherein the aryl is phenyl, biphenyl, naphthalene, anthracene, or a combination thereof.

19. The compound of claim 14, wherein the sugar has formula IIa or IIIb:

20. A compound selected from the group consisting of:

21. A compound comprising the following formula:

44. The method of cell 35, wherein the cell expresses Siglec-8 and the Siglec ligand compound has the following formula:

45. The method of claim 44, wherein the cell is an eosinophil.

48. A method for killing a B cell that expresses CD22 comprising administering to the B cell a Siglec ligand compound and exposing the cell to complement factors, wherein the Siglec ligand compound is selected from the group consisting of:

49. A method of treating an eosinophil-related disease or condition in a mammal comprising administering to the mammal an effective amount of a compound of the formula:

50. The method of claim 49, wherein the disease or condition is asthma, rhinitis or eczema.

51. A multivalent Carrier-Siglec ligand complex comprising a Siglec ligand, a neutral saccharide and a multivalent Carrier, wherein the Siglec ligand is a compound of formula IIa or IIb:

52. The multivalent complex of claim 51, wherein the Carrier is a polymer, protein, antibody, antigen, multi-subunit protein, protein complex, glycan, lipid, liposome, nucleic acid, polysaccharide, glycoprotein, glycolipid, lipoprotein, bead or solid support.