Decoy influenza therapies

• US 8,815,522 B2
• Filed: 01/02/2009
• Issued: 08/26/2014
• Est. Priority Date: 01/03/2008
• Status: Active Grant

• Current Assignee: Massachusetts Institute of Technology
• Original Assignee: Massachusetts Institute of Technology
• Inventors: Sasisekharan, Ram, Viswanathan, Karthik, Chandrasekaran, Aarthi, Raman, Rahul, Srinivasan, Aravind, Raguram, S., Sasisekharan, Viswanathan
• Primary Examiner(s): Kosson, Rosanne
• Application Number:
  US12/348,265
• Publication Number:
  US 20100004195A1
• Time in Patent Office: 2,062 Days
• Field of Search: 435/7.2, 514/61, 514/54, 536/123.1
• US Class Current: 435/7.2
• CPC Class Codes: A61P 31/16 for influenza or rhinoviruses
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: G01N 2333/11 Orthomyxoviridae, e.g. influenza virus
• CPC Class Codes: G01N 2400/10 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: G01N 33/56983 Viruses

Abstract

The present invention provides umbrella-topology glycan decoys. The present invention provides systems and methods treating influenza infection utilizing inventive umbrella-topology glycan decoys. The present invention provides methods for identifying novel umbrella-topology glycan decoys.

Claims

1. A method for identifying agents useful in the treatment of influenza infection, the method comprising steps of:

(a) providing an influenza hemagglutinin (HA), wherein the HA is characterized in that it preferentially binds to umbrella-topology glycans over cone-topology glycans such that it interacts with HA receptor glycans preferentially found in the human upper respiratory tract;

(b) providing a collection of candidate umbrella-topology decoys, which candidate umbrella-topology decoys comprise glycan moieties that share an oligosaccharide branch length characteristic with at least one umbrella-topology glycan selected from the group consisting of;

(i) those depicted in FIG. 6,

(ii) those depicted in FIG. 7,

(iii) those depicted in FIG. 9,

(iv) α

2-6 sialylated glycans with length of at least a tetrasaccharide and characterized by one or more of θ

<

100°

, φ

angle of Neu5Acα

2-6Gal linkage of around −

60, and multiple lactosamine units, and

(v) combinations thereof;

(c) contacting the influenza HA with one or more candidate umbrella-topology decoy from the collection;

(d) determining that at least one of the candidate umbrella-topology decoys from the collection competes the binding interaction between the influenza HA and one or more of the HA receptor glycans preferentially found on human upper respiratory tract, so that the candidate umbrella-topology decoy is useful in the treatment of influenza infection.

2. The method of claim 1, wherein the collection of candidate umbrella-topology decoys are sialylated glycans, wherein the sialylated glycans are provided as glycolipids or glycoproteins.

3. The method of claim 1, wherein the collection of candidate umbrella-topology decoys comprise one or more molecules physically associated with a carrier moiety.

4. The method of claim 3, wherein the one or more molecules physically associated with a carrier are physically associated by way of a covalent bond with the carrier moiety.

5. The method of claim 3, wherein the carrier moiety is selected from the group consisting of a peptide, glycopeptide, nucleic acid, small molecule, lipid, fatty acid group, carbohydrate, cell, virus, and particle.

6. The method of claim 1, wherein the at least one candidate decoy that mimics umbrella-topology glycans is characterized by a three-dimensional structure that interacts with one or more of HA amino acids at positions selected from the group consisting of 136, 137, 145, 153, 155, 156, 159, 186, 187, 189, 190, 192, 193, 194, 196, 222, 225, 226, 228, and combinations thereof, relative to a reference wild type H3 HA corresponding to the polypeptide of SEQ ID NO. 6.

7. The method of claim 1, wherein the at least one candidate decoy that mimics umbrella-topology glycans is characterized by a three-dimensional structure that interacts with one or more of HA amino acid acids at positions selected from the group consisting of 156, 159, 189, 192, 193, 196, and combinations thereof, relative to a reference wild type H3 HA corresponding to the polypeptide of SEQ ID NO. 6.

8. The method of claim 1, wherein the influenza HA is selected from the group consisting of:

H1, H2, and H3 HA.

9. The method of claim 1, wherein the influenza HA is from a strain that is able to infect humans.

10. The method of claim 1, further comprising contacting the influenza HA with a positive control that is known to bind the influenza HA.

11. The method of claim 10, wherein the positive control is an umbrella-topology glycan.

12. The method of claim 1, wherein some or all of the candidate umbrella-topology decoys share one or more features with umbrella-topology glycans that are not shared with cone-topology glycans, such feature being selected from the group consisting of solvent accessibility, surface area, glycosidic torsion angles, conformational energy distribution, linkage, oligosaccharide branch length, amino acid residue contact sites, structural topology, binding characteristics, φ

values, ψ

values, θ

values, ω

angles, and combinations thereof.

13. The method of claim 1, wherein the HA receptor glycans preferentially found on human upper respiratory tract are at least 2-fold more likely to be found on human upper respiratory epithelial cells than on other cells.

14. The method of claim 1, wherein the HA receptor glycans preferentially found on human upper respiratory tract are at least 10-fold more likely to be found on human upper respiratory epithelial cells than on other cells.

15. The method of claim 1, wherein the HA receptor glycans preferentially found on human upper respiratory tract are at least 50-fold more likely to be found on human upper respiratory epithelial cells than on other cells.

16. The method of claim 1, wherein the at least one umbrella-topology glycan comprises at least one oligosaccharide of the following structure:

Neu5Acα

2-6Sug1-Sug2-Sug3

where;

(a) Neu5Acα

2-6 may be at the non-reducing end;

(b) Sug1;

(i) is or comprises hexose or hexosamine in α

or β

configuration;

(ii) does not include any sugars other than Neu5Acα

2-6 attached to any of its non-reducing positions except when Sug1 is GalNAα

2-6 that is O-linked to the glycoprotein; and

(iii) optionally comprises at least one non-sugar moiety attached to a non-reducing position;

(c) at least one of Sug2 and Sug3 is or comprises;

(i) hexose or hexosamine in a or 0 configuration; and

(ii) optionally one or more sugar or non-sugar moiety attached to a non-reducing position;

(d) linkage between any two sugars in the oligosaccharide apart from Neu5Acα

2-6 linkage is selected from the group consisting of;

1-2, 1-3, 1-4, and 1-6.

17. The method of claim 1, wherein the at least one umbrella-topology glycan comprises at least one oligosaccharide of the following structure:

Neu5Acα

2-6Sug1-Sug2-Sug3-Sug4-

where;

(a) Neu5Acα

2-6 may be at the non-reducing end;

(b) Sug1;

(i) is or comprises hexose or hexosamine in a or 0 configuration;

(ii) does not include any sugars other than Neu5Acα

2-6 attached to any of its non-reducing positions; and

(iii) optionally comprises at least one non-sugar moiety attached to a non-reducing position;

(c) at least one of Sug2, Sug3, and Sug4 is or comprises;

(i) hexose or hexosamine in α

or β

configuration; and

(ii) optionally one or more sugar or non-sugar moiety attached to a non-reducing position;

(d) linkage between any two sugars in the oligosaccharide apart from Neu5Acα

2-6 linkage is selected from the group consisting of;

1-2, 1-3, 1-4, and 1-6.

18. The method of claim 16 or 17, wherein the hexose is selected from the group consisting of:

Gal and Glc.

19. The method of claim 16 or 17, wherein the hexosamine is selected from the group consisting of:

GlcNAc and GalNAc.

20. The method of claim 16 or 17, wherein the hexose or hexosamine is in the β

configuration.

21. The method of claim 16, wherein one or more of Sug1, Sug2, and Sug3 comprises a non-sugar moiety attached to a non-reducing position.

22. The method of claim 17, wherein one or more of Sug1, Sug2, Sug3, and Sug4 comprises a non-sugar moiety attached to a non-reducing position.

23. The method of claim 21 or 22, wherein Sug1 comprises a non-sugar moiety attached to non-reducing position 6.

24. The method of claim 21 or 22, wherein the non-sugar moiety is selected from the group consisting of:

sulfate, phosphate, guanidinium, amine, N-acetyl, and combinations thereof.

25. The method of claim 16, wherein the one or more of Sug2 and Sug3 comprises a sugar moiety attached to a non-reducing position.

26. The method of claim 17, wherein the one or more of Sug2, Sug3, and Sug4 comprises a sugar moiety attached to a non-reducing position.

27. The method of claim 24 or 26, wherein the one or more sugar moiety is Fuc.

28. The method of claim 16 or 17, wherein the at least one linkage between two sugars in the oligosaccharide apart from Neu5Acα

2-6 linkage is selected from the group consisting of;

1-3 and 1-4.

29. The method of claim 16, wherein the at least one umbrella-topology glycan comprises Neu5Acα

2-6 linked to GalNAcα

that is O-linked to the glycoprotein and additional sugars are linked to the non-reducing end of GalNAcα

.

30. The method of claim 28, wherein the at least one umbrella-topology glycan is Neu5Acα

2-6(Neu5Acα

2-3Galβ

1-3)GalNAcα

-.

31. The method of claim 29, wherein the at least one umbrella-topology glycan is Neu5Acα

2-6(Galβ

1-3)GalNAcα

-.

32. The method of claim 17, wherein the at least one umbrella-topology glycan comprises a oligosaccharide structure without Sug4 such as 6′

SLN that is attached to O-linked core GalNAc.

33. The method of claim 32, wherein the at least one umbrella-topology glycan is (Neu5Acα

2-6Galβ

1-4GlcNAc)β

1-3/6(GalNAcα

-O-Ser).

34. The method of claim 1, wherein the at least one umbrella-topology glycan is selected from the group consisting of:

Neu5Acα

2-6Galβ

1-4GlcNAcβ

1-3Galβ

1-4GlcNAc-, Neu5Acα

2-6GalNAcβ

1-4GlcNAcβ

1-3GalNAcβ

1-4GlcNAc-, Neu5Acα

2-6GlcNAcβ

1-3Galβ

1-3/4GlcNAc-, Neu5Acα

2-6Galβ

1-4GlcNAcβ

1-3Galβ

1-3GalNAcα

, Neu5Acα

2-6Galβ

1-4GlcNAcβ

1-3Galβ

1-4GlcNAcβ

1-3Galβ

1-3GalNAcα

, Neu5Acα

2-6GalNAcβ

1-4GlcNAcβ

1-3Galβ

1-3GalNAcα

, Neu5Acα

2-6GalNAcβ

1-4GlcNAcβ

1-3GalNAcβ

1-4GlcNAcβ

1-3Galβ

1-3GalNAcα

, Neu5Acα

2-6 GalNAcα

-β

1-3Galα

2-3Neu5Ac, Neu5Acα

2-6Galβ

1-4GlcNAcβ

1-3/6GalNAcα

, Neu5Acα

2-6Galβ

1-4GlcNAcβ

1-3Galβ

1-4GlcNAcβ

1-3/6GalNAcα

, Neu5Acα

2-6GalNAcβ

1-4GlcNAcβ

1-3/6GalNAcα

, Neu5Acα

2-6GalNAcβ

1-4GlcNAcβ

1-3GalNAcβ

1-4GlcNAcβ

1-3/6GalNAcα

, Neu5Acα

2-6Galβ

1-4GlcNAcβ

1-6GalNAcα

-β

1-3Galα

2-3Neu5Ac, Neu5Acα

2-6Galβ

1-4GlcNAcβ

1-3/6GalNAcα

-β

1-3/6GlcNAcβ

1-4Galα

2-3/6Neu5Ac, Neu5Acα

2-6GlcNAcβ

1-3Galβ

1-4GlcNAcβ

1-3Galβ

1-3GalNAc, Neu5Acα

2-6GlcNAcβ

1-3Galβ

1-3GlcNAcβ

1-3/6GalNAc, Neu5Acα

2-6GlcNAcβ

1-3Galβ

1-4GlcNAcβ

1-3/6GalNAc, Neu5Acα

2-6Galβ

1-3GalNAcβ

1-4Galα

1-3Galβ

1-4Glc, Neu5Acα

2-6Galβ

1-4GlcNAcβ

1-3Galβ

1-4Glc, Neu5Acα

2-6Galβ

1-3GalNAcβ

1-3Galα

1-4Galβ

1-4Glc, Neu5Acα

2-6Galβ

1-3GlcNAcβ

1-3Galβ

1-4Glc, Neu5α

2-6Galβ

1-4GlcNAcβ

1-3GalNAcα

, Neu5Acα

2-6Galβ

1-4GlcNAcβ

1-3Galβ

1-3GalNAcα

, Neu5Acα

2-6GalNAc(β

1-3Gal-)β

1-4Galβ

1-4Glc, Neu5Acα

2-6GalNAc(β

1-3Gal-)β

1-3Galα

1-4Galβ

1-4Glc, and combinations thereof.

35. The method of claim 1, wherein the cone-topology glycans are as set forth in FIG. 8.

36. The method of claim 1, wherein the cone-topology glycans are selected for the group consisting of:

Neu5Acα

2-3Galβ

1-3GlcNAc, Neu5Acα

2-3/6Galβ

1-4GlcNAc, Neu5Acα

2-3Galβ

1-3GalNAc-, and combinations thereof.