Adiponectin and uses thereof
View Patent ↗The invention provides methods and reagents for regulation of metabolic events, such as those mediated by adiponectin and adiponectin agonists. The invention also provides screening assays for identification of biologically active agents, diagnostic and therapeutic agents, and other methods and reagents.
1. A composition which comprises a pharmaceutically acceptable carrier, and substantially pure glycosylated human adiponectin produced in transformed mammalian cells in an amount effective for treating inflammation of the liver, wherein one or more lysine residues at positions 65, 68, 77, and 101 is hydroxylated and comprise a glucosylgalactosyl moiety, a glucosylglucosyl moiety, a galactosylglucosyl moiety or a galactosyl moiety.
2. The composition of claim 1 further comprises an antimicrobial agent.
3. The composition of claim 1 wherein the amount is effective to elicit a plasma adiponectin concentration from about 1 μg/mL to about 20 μg/mL.
4. The composition of claim 1 wherein the amount is effective to elicit a plasma adiponectin concentration from about 1.9 μg/mL to about 17 μg/mL.
5. The composition of claim 1 wherein the glycosylated human adiponectin is at least about 50% pure.
6. The composition of claim 1 wherein the glycosylated human adiponectin is at least about 80% pure.
7. The composition of claim 1 wherein the glycosylated human adiponectin is at least about 90% pure.
8. The composition of claim 1 wherein the glycosylated human adiponectin is at least about 95% pure.
9. The composition of claim 1 wherein the glycosylated human adiponectin is at least about 99% pure.
10. The composition of claim 1 wherein the lysine residues at positions 65, 68, 77, and 101 comprise a glucosylgalactosyl moiety, a glucosylglucosyl moiety, a galactosylglucosyl moiety, or a galactosylgalactosyl moiety.
11. The composition of claim 1 wherein one or more of the lysine residues at positions 65, 68, 77, and 101 comprise a glucosylgalactosyl moiety or a galactosylglucosyl moiety.
12. The composition of claim 1 wherein the lysine residues at positions 65, 68, 77, and 101 comprise a glucosylgalactosyl moiety or a galactosylglucosyl moiety.
13. The composition of claim 1 wherein one or more of the residues at positions 65, 68, 77, and 101 is glucosyl-α-1-2-galactosyl-O-lysine.
14. The composition of claim 1 wherein the human adiponectin comprises a glycosylgalactosyl moiety attached to the lysine residues at positions 65, 68, 77, and 101.
15. The composition of claim 1 wherein the human adiponectin comprises a lysine with a glucosylgalactosyl moiety or a galactosylglucosyl moiety at positions 65, 68, 77, and 101.
16. The composition of claim 1 wherein each of the human adiponectin lysine residues 65, 68, 77 and 101 is glucosyl-α-1-2-galactosyl-O-lysine and residue 91 is hydroxyproline.
17. The composition of claim 1 wherein the adiponectin has the amino acid sequence of a naturally occurring human adiponectin.
18. The composition of claim 1 wherein the amino acid residue at position 91 of the glycosylated human adiponectin is hydroxyproline.
19. The composition of claim 1 wherein the human adiponectin is a mixture of one or more of isoforms 3, 4, 5 and 6.
20. The composition of claim 1 which is substantially free of non-glycosylated adiponectin.
21. The composition of claim 1 which is substantially free of isoform 1.
22. The composition of claim 1 which is substantially free of isoform 2.
23. The composition of claim 1 wherein the composition comprises more than one isoform of glycosylated human adiponectin.
24. The composition of claim 1 wherein the composition comprises one or more of glycosylated human adiponectin isoforms 3-8.
25. The composition of claim 19 wherein human adiponectin isoform 3 is the predominant adiponectin isoform.
26. The composition of claim 1 wherein human adiponectin isoform 4 is the predominant adiponectin isoform.
27. The composition of claim 1 wherein human adiponectin isoform 5 is the predominant adiponectin isoform.
28. The composition of claim 1 wherein human adiponectin isoform 6 is the predominant adiponectin isoform.
29. The composition of any of claims 25 - 27 or 28 wherein said isoform is at least about 50% the glycosylated human adiponectin in the composition.
30. The composition of any of claims 25 - 27 or 28 wherein said isoform is at least about 80% the glycosylated human adiponectin in the composition.
31. The composition of any of claims 25 - 27 or 28 wherein said isoform is at least about 90% the glycosylated human adiponectin in the composition.
32. The composition of any of claims 25 - 27 or 28 wherein said isoform is at least about 95% the glycosylated human adiponectin in the composition.
33. The composition of any of claims 25 - 27 or 28 wherein said isoform is at least about 98% the glycosylated human adiponectin in the composition.
34. The composition of claim 1 wherein the composition is formulated in a manner suitable for parenteral administration.
35. The composition of claim 1 wherein the composition is formulated in a manner suitable for intravenous administration.
36. The composition of claim 1 wherein the composition is formulated in a manner suitable for subcutaneous administration.
37. The composition of claim 1 , further comprising a buffer.
38. The composition of claim 1 , further comprising a tonicity agent.
39. The composition of claim 2 , wherein the antimicrobial agent is present in a bacteriostatic or fungistatic concentration.
40. The composition according to claim 10 wherein the glycosylated lysine residues at 65, 68, 77 and 101 comprise glucosylgalactosyl moieties.
41. The composition according to claim 1 wherein the adiponectin comprises glycosylated lysine residues at positions 65, 68, 77 and 101 glycosylated with a glycosylgalactosyl moiety.
42. The composition of claim 1 wherein each of the glycosylated lysine residues at 65, 68, 77 and 101 is glucosyl-α, 1,2-galactosyl-O-lysine.
43. The composition of claims 40 , 41 and 42 which is substantially free of non-glycosylated adiponectin.
44. The composition of claim 40 wherein the glycosylated human adiponectin is recombinant.
45. The composition of claim 40 wherein the amount is effective to elicit a plasma adiponectin concentration from about 1 μg/mL to about 20 μg/mL.
46. The composition of claim 40 wherein the amount is effective to elicit a plasma adiponectin concentration from about 1.9 μg/mL to about 17 μg/mL.
47. The composition of claim 40 wherein the glycosylated human adiponectin is at least about 50% pure.
48. The composition of claim 40 wherein the glycosylated human adiponectin is at least about 80% pure.
49. The composition of claim 40 wherein the glycosylated human adiponectin is at least about 90% pure.
50. The composition of claim 40 wherein the glycosylated human adiponectin is at least about 95% pure.
51. The composition of claim 40 wherein the glycosylated human adiponectin is at least about 99% pure.
52. The composition of claim 40 wherein the composition is formulated in a manner suitable for parenteral administration.
53. The composition of claim 40 wherein the composition is formulated in a manner suitable for intravenous administration.
54. The composition of claim 40 wherein the composition is formulated in a manner suitable for subcutaneous administration.
55. The composition of claim 40 , further comprising a buffer.
56. The composition of claim 40 , further comprising a tonicity agent.
57. The composition of claim 40 further comprises an antimicrobial agent, wherein the antimicrobial agent is present in a bacteriostatic or fungistatic concentration.