APTAMERS FOR MEASURING LIPOPROTEIN LEVELS

• US 20200131515A1
• Filed: 07/06/2018
• Published: 04/30/2020
• Est. Priority Date: 07/07/2017
• Status: Active Grant

• Current Assignee: InnaMed, Inc.
• Original Assignee: InnaMed, Inc.
• Inventors: INAPURI, Eshwar, SINGH, Anup, PENNER, Gregory Allen
• Granted Patent: US 11,505,799 B2
• Time in Patent Office: Days
• CPC Class Codes: C12N 15/115 Aptamers, i.e. nucleic acids binding a target molecule specifically and with high affinity without hybridising therewith ; Nucleic acids binding to non-nucleic acids, e.g. aptamers
• CPC Class Codes: C12N 2310/113 targeting other non-coding nucleic acids, e.g. antagomirs
• CPC Class Codes: C12N 2310/16 Aptamers
• CPC Class Codes: C12N 2320/10 in screening processes
• CPC Class Codes: G01N 33/48 Biological material, e.g. blood, urine G01N33/02, G01N33/26, G01N33/44, G01N33/46 take precedence; Haemocytometers counting blood corpuscules distributed over a surface by scanning the surface G06M11/02
• CPC Class Codes: G01N 33/54313 the carrier being characterised by its particulate form
• CPC Class Codes: G01N 33/92 involving lipids, e.g. cholesterol , lipoproteins, or their receptors steroid hormones G01N33/743

Abstract

Provided herein are aptamers for binding lipoproteins, systems for binding lipoproteins, and methods of detecting lipoproteins in a sample. The aptamers are useful for detecting the levels of lipoproteins in a biological sample and selectively detecting LDL particles in the presence of HDL particles. The aptamers can also be used as therapeutic agents against various diseases.

Claims

1. An aptamer with high affinity to LDL particles and high selectivity for LDL particles over HDL particles.

2. The aptamer of claim 1, wherein the aptamer is highly selective for L-LDL particles over H-LDL particles.

3. The aptamer of claim 1 or 2, wherein the aptamer comprises SEQ. ID NO. 1:

ACCTCGATTTTATATTATTTCGCTTACCAACAACTGCAGA.

4. The aptamer of claim 1 or 2, wherein the aptamer comprises SEQ. ID NO. 2:

GTCAGCTTCTTCTATATAATTCACCTTGCCTCTCGTTCCT.

5. The aptamer of claim 1 or 2, wherein the aptamer comprises SEQ. ID NO. 3:

AACCTTAATTTACCCTTGTTTTATCATTTCCATCATAGCT.

6. The aptamer of claim 1 or 2, wherein the aptamer comprises SEQ. ID NO. 4:

AAACCATTCTTGTCTCGCATTTTTCTCTTTTATATTATTT.

7. The aptamer of claim 1 or 2, wherein the aptamer comprises SEQ. ID NO. 5:

CACGCGCATAACCCTTTTTTCACCTGCCCTTTTAACTAAT.

8. The aptamer of claim 1 or 2, wherein the aptamer comprises SEQ. ID NO. 6:

GCCATAAGATGACGACATTCATCACCAAACCAACCTCAGC.

9. The aptamer of claim 1 or 2, wherein the aptamer comprises SEQ. ID NO. 7:

GCCGGGTGATTGAAAAGCAGATTACCGATCATCCAATAAA.

10. The aptamer of claim 1 or 2, wherein the aptamer consists of SEQ. ID NO. 8:

AACTACATGGTATGTGGTGAACTACCTCGATTTTATATTATTTCGCTTACCAACAACT GCAGAGACGTACAATGTACCC.

11. The aptamer of claim 1 or 2, wherein the aptamer consists of SEQ. ID NO. 9:

AACTACATGGTATGTGGTGAACTGTCAGCTTCTTCTATATAATTCACCTTGCCTCTCG TTCCTGACGTACAATGTACCC.

12. The aptamer of claim 1 or 2, wherein the aptamer consists of SEQ. ID NO. 10:

AACTACATGGTATGTGGTGAACTAACCTTAATTTACCCTTGTTTTATCATTTCCATCA TAGCTGACGTACAATGTACCC.

13. The aptamer of claim 1 or 2, wherein the aptamer consists of SEQ. ID NO. 11:

AACTACATGGTATGTGGTGAACTAAACCATTCTTGTCTCGCATTTTTCTCTTTTATAT TATTTGACGTACAATGTACCC.

14. The aptamer of claim 1 or 2, wherein the aptamer consists of SEQ. ID NO. 12:

AACTACATGGTATGTGGTGAACTCACGCGCATAACCCTTTTTTCACCTGCCCTTTTAA CTAATGACGTACAATGTACCC.

15. The aptamer of claim 1 or 2, wherein the aptamer consists of SEQ. ID NO. 13:

AACTACATGGTATGTGGTGAACTGCCATAAGATGACGACATTCATCACCAAACCAA CCTCAGCGACGTACAATGTACCC.

16. The aptamer of claim 1 or 2, wherein the aptamer consists of SEQ. ID NO. 14:

AACTACATGGTATGTGGTGAACTGCCGGGTGATTGAAAAGCAGATTACCGATCATCC AATAAAGACGTACAATGTACCC.

17. A reagent comprising the aptamer according to any one of claims 1-16.

18. A kit comprising the aptamer according to any one of claims 1-16.

41. A method of detecting, identifying, and/or quantifying the concentration of lipoprotein particles in a sample, comprising exposing the sample to an aptamer or a plurality of aptamers according to any one of claims 1-16, measuring the signal generated by the sample, and analyzing the signal to quantify the concentration of lipoprotein particles in the sample, wherein the signal changes in proportion to the concentration of lipoprotein particles.

42. The method of claim 41, wherein the aptamer or plurality of aptamers selectively binds to LDL particles in the presence of HDL particles.

43. The method of claim 41 or 42, wherein the aptamer or plurality of aptamers selectively binds to L-LDL particles in the presence of H-LDL particles.

44. The method of any one of claims 41 to 43, wherein the aptamer or plurality of aptamers is attached to a surface.

45. The method of claim 44, wherein the surface is a gold chip.

46. The method of any one of claims 41 to 45, wherein the sample comprises blood serum, whole blood, nasal aspirates, saliva, urine, feces, cell lysate, dialysis sampling, tissue biopsy, cell media, or a combination thereof.

47. The method of claim 46, wherein the sample comprises blood serum.

48. The method of any one of claims 41 to 47, wherein the sample contains low-density lipoprotein (LDL) and high-density lipoprotein (HDL) particles.

49. The method of any one of claims 41 to 48, wherein the bound lipoprotein particle is an LDL particle.

50. The method of claim 49, wherein the LDL particle is an H-LDL particle.

51. The method of claim 49, wherein the LDL particle is an L-LDL particle.

52. A method of modulating the activity of a protein in a cell, comprising contacting the protein with the aptamer, reagent, or kit of any one of claims 1 to 18.

53. A method of treating or preventing a disease or disorder in a mammal, comprising administering to the mammal the aptamer, reagent, or kit of any one of claims 1 to 18.

19. A system comprising a plurality of aptamers, wherein the aptamers have a high affinity to LDL particles and high selectivity for LDL particles over HDL particles.

20. The system of claim 19, wherein the plurality of aptamers is immobilized onto a surface.

21. The system of claim 19, wherein a plurality of antisense oligonucleotides is immobilized onto a surface and the plurality of aptamers is annealed to the plurality of antisense oligonucleotides.

22. The system of claim 21, wherein the surface is a gold-, carbon-, or silicon-containing chip.

23. The system of claim 22, wherein the chip is coated with a gold-containing film.

24. The system of claim 23, wherein the film is patterned.

25. The system of claim 23 or 24, wherein the film has a thickness of between about 0.2 mm and 2 mm.

26. The system of any one of claims 19 to 25, wherein the plurality of aptamers is highly selective for L-LDL particles over H-LDL particles.

27. The system of any one of claims 19 to 26, wherein the plurality of aptamers comprises SEQ. ID NO. 1.

28. The system of any one of claims 19 to 26, wherein the plurality of aptamers comprises SEQ. ID NO. 2.

29. The system of any one of claims 19 to 26, wherein the plurality of aptamers comprises SEQ. ID NO. 3.

30. The system of any one of claims 19 to 26, wherein the plurality of aptamers comprises SEQ. ID NO. 4.

31. The system of any one of claims 19 to 26, wherein the plurality of aptamers comprises SEQ. ID NO. 5.

32. The system of any one of claims 19 to 26, wherein the plurality of aptamers comprises SEQ. ID NO. 6.

33. The system of any one of claims 19 to 26, wherein the plurality of aptamers comprises SEQ. ID NO. 7.

34. The system of any one of claims 19 to 26, wherein the plurality of aptamers comprises SEQ. ID NO. 8.

35. The system of any one of claims 19 to 26, wherein the plurality of aptamers comprises SEQ. ID NO. 9.

36. The system of any one of claims 19 to 26, wherein the plurality of aptamers comprises SEQ. ID NO. 10.

37. The system of any one of claims 19 to 26, wherein the plurality of aptamers comprises SEQ. ID NO. 11.

38. The system of any one of claims 19 to 26, wherein the plurality of aptamers comprises SEQ. ID NO. 12.

39. The system of any one of claims 19 to 26, wherein the plurality of aptamers comprises SEQ. ID NO. 13.

40. The system of any one of claims 19 to 26, wherein the plurality of aptamers comprises SEQ. ID NO. 14.