Antisense and pretargeting optical imaging

• US 20110158913A1
• Filed: 01/18/2008
• Published: 06/30/2011
• Est. Priority Date: 01/19/2007
• Status: Abandoned Application

• Current Assignee: Kayoko Nakamura, University Of Massachusetts
• Original Assignee: University Of Massachusetts
• Inventors: Liu, Xinrong, Wang, Yi, He, Jiang, Zhang, Surong, Rusckowski, Mary, Hnatowich, Don, Nakamura, Kayoko
• Application Number:
  US12/009,452
• Publication Number:
  US 20110158913A1
• Time in Patent Office: Days
• US Class Current: 424/9.6
• CPC Class Codes: A61K 49/0032 Methine dyes, e.g. cyanine dyes
• CPC Class Codes: A61K 49/0054 Macromolecular compounds, i.e. oligomers, polymers, dendrimers
• CPC Class Codes: G01N 33/542 with steric inhibition or signal modification, e.g. fluorescent quenching
• CPC Class Codes: G01N 33/574 for cancer
• CPC Class Codes: G01N 33/582 with fluorescent label

Abstract

The present invention relates, in part, to detectably labeled oligomer duplexes and their use in optical imaging, including, in vivo optical imaging. The invention includes methods of optical imaging including in vivo pretargeting methods and in vivo antisense optical imaging methods.

Claims

1. A method of optically imaging a target entity in a subject, the method comprising

(a) administering to the subject a binding molecule that comprises an oligomer moiety and specifically binds to the target entity,

(b) contacting the binding molecule with a quenched first linear oligomer duplex comprising a detectably labeled oligomer, wherein the detectably labeled oligomer is hybridized to a quenching oligomer;

the detectable label is quenched unless the detectably labeled oligomer and the quenching oligomer dissociate; and

the detectably labeled oligomer has a higher affinity to form a linear duplex with the oligomer moiety of the binding molecule than to form a linear duplex with the quenching oligomer, and

(c) detecting the presence of unquenched detectable label in the subject, wherein the presence of unquenched detectable label indicates that the detectably labeled oligomer has dissociated from the quenching oligomer and hybridized to the oligomer of the binding molecule to form a second linear oligomer duplex, permitting optical imaging of the target entity in the subject.

2. The method of claim 1, wherein one or more of the oligomers are phosphodiester, phosphorothioate, peptide nucleic acid (PNA), locked nucleic acid (LNA), and/or phosphorodiamidate morpholino (MORF) oligomers.

3. The method of claim 1, wherein the detectable label is a fluorescent or bioluminescent label.

4. The method of claim 3, wherein the fluorescent label is a Cy5.5 emitter.

5. The method of claim 1, wherein the quenching oligomer comprises a quenching moiety.

6. The method of claim 5, wherein the quenching moiety is BHQ3 or Iowa black.

7. The method of claim 1, wherein the detectable label is detected in vivo.

8. The method of claim 1, wherein the detectable label is detected in real time.

9. The method of claim 1, wherein the first linear oligomer duplex comprises two oligomers that are not both phosphodiester, phosphorothioate, peptide nucleic acid (PNA), locked nucleic acid (LNA), or phosphorodiamidate morpholino (MORF) oligomers.

10. The method of claim 1, wherein the oligomer moiety is a single-stranded nucleic acid moiety.

11. The method of claim 1, wherein the binding molecule is an antibody or antigen-binding fragment thereof.

12. The method of claim 1, wherein the target entity is a polypeptide, nucleic acid, polysaccharide or lipid molecule.

13. The method of claim 1, wherein the target entity is a cell.

14. The method of claim 13, wherein the cell is a cancer cell.

15. The method of claim 1, wherein the subject is human.

16. The method of claim 1, wherein presence of the specific target entity is associated with a disease or disorder.

17. The method of claim 16, wherein the disease is cancer.

18. The method of claim 1, wherein the optical imaging of the target entity in the subject is diagnostic for a disease or disorder in the subject.

19. The method of claim 18, wherein the disease is cancer.

20. A method of optically imaging a target entity in a subject, the method comprising

(a) administering to the subject a linear oligomer duplex comprising a detectably labeled oligomer and a quenching oligomer, wherein the detectable label is quenched by the quenching oligomer unless the detectably labeled oligomer and quenching oligomer of the duplex dissociate, and wherein the detectably labeled oligomer has a higher affinity to form a duplex with a specific target nucleic acid than to form a duplex with the quenching oligomer, and

(b) detecting the presence of unquenched detectable label in the subject, wherein the presence of unquenched detectable label in the subject indicates that the detectably labeled oligomer has formed a duplex with the specific target nucleic acid, permitting optical imaging of the target nucleic acid in the subject.

21. The method of claim 20, wherein one or more of the oligomers are phosphodiester, phosphorothioate, peptide nucleic acid (PNA), locked nucleic acid (LNA), and/or phosphorodiamidate morpholino (MORF) oligomers.

22. The method of claim 20, wherein the detectable label is a fluorescent or bioluminescent label.

23. The method of claim 22, wherein the fluorescent label is a Cy5.5 emitter.

24. The method of claim 20, wherein the quenching oligomer comprises a quenching moiety.

25. The method of claim 24, wherein the quenching moiety is BHQ3 or Iowa black.

26. The method of claim 20, wherein the detectable label is detected in vivo.

27. The method of claim 20, wherein the detectable label is detected in real time.

28. The method of claim 20, wherein the specific target nucleic acid is an mRNA.

29. The method of claim 20, wherein the detectably labeled oligomer is an antisense sequence to the specific target nucleic acid.

30. The method of claim 20, wherein expression of the specific target nucleic acid is associated with a disease or disorder.

31. The method of claim 30, wherein the disease is cancer.

32. The method of claim 20, wherein the subject is human.

33. The method of claim 20, wherein the optical imaging of the target nucleic acid in the subject is diagnostic for a disease or disorder in the subject.

34. The method of claim 33, wherein the disease is cancer.

35. The method of claim 20, wherein the detectably labeled oligomer and the quenching oligomer are not both phosphodiester, phosphorothioate, peptide nucleic acid (PNA), locked nucleic acid (LNA), or phosphorodiamidate morpholino (MORF) oligomers.