Compositions, probes and conjugates and uses thereof

• US 8,003,771 B2
• Filed: 03/01/2010
• Issued: 08/23/2011
• Est. Priority Date: 05/22/2006
• Status: Active Grant

• Current Assignee: Gen-Probe Incorporated (Hologic Incorporated)
• Original Assignee: Third Wave Technologies Incorporated (Hologic Incorporated)
• Inventors: Skrzypczynski, Zbigniew, Wayland, Sarah R
• Primary Examiner(s): Riley; Jezia
• Application Number:
  US12/714,983
• Publication Number:
  US 20100152431A1
• Time in Patent Office: 540 Days
• Field of Search: 536/23.1, 536/25.3, 552/6, 514/383, 435/116, 435/129
• US Class Current: 536/23.1
• CPC Class Codes: A61K 49/0039 Coumarin dyes
• CPC Class Codes: A61K 49/0054 Macromolecular compounds, i.e. oligomers, polymers, dendrimers
• CPC Class Codes: C07H 21/00 Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids

Abstract

The present invention relates to compositions useful as probes and in other applications and methods of their use. In some embodiments, nucleotides are prepared and functionalized with dyes. In some embodiments a first molecule is functionalized with an alkynyl group, a second molecule is functionalized with an azide group, and said first and second molecules are mixed under conditions to form a conjugate with a 1,2,3-triazol group. In further embodiments, a nucleotide is functionalized with an alkynyl group, a dye is functionalized with an azide group, and mixing the nucleotide and the dye forms a conjugate capable of emitting light.

Claims

1. A composition comprising an unreacted compound functioning to react with an azide to form a reacted compound comprising a 1,2,3-triazol group, said unreacted compound having the following formula:

2. The composition of claim 1, wherein said unreacted compound comprises the structure:

3. The composition of claim 1, wherein at least one of R¹, R², and R³ is an unsaturated alkyl group.

4. The composition of claim 1, wherein at least one of R¹, R², and R³ is a saturated alkyl group.

5. The composition of claim 1, wherein at least one of R¹, R², and R³ is a substituted alkyl group.

6. The composition of claim 1, wherein at least one of R¹, R², and R³ is an unsubstituted alkyl group.

7. The composition of claim 1, wherein at least one of R¹, R², and R³ is a straight-chain alkyl group.

8. The composition of claim 7, wherein said straight-chain alkyl group is selected from the group consisting of methyl, ethyl, n-propyl, n-butyl, n-pentyl, n-hexyl, n-septyl, n-octyl, and n-nonyl groups.

9. The composition of claim 1, wherein at least one of R¹, R², and R³ is a branched alkyl group.

10. The composition of claim 9, wherein said branched alkyl group is selected from the group consisting of isopropyl, sec-butyl, isobutyl, tert-butyl, and isopentyl alkyl groups.

11. The composition of claim 1, wherein at least one of R¹, R², and R³ is a cyclic alkyl group.

12. The composition of claim 11, wherein said cyclic alkyl group is homocyclic.

13. The composition of claim 11, wherein said cyclic alkyl group is heterocyclic.

14. The composition of claim 11, wherein said cyclic alkyl group is selected from the group consisting of cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclopentenyl and cyclohexenyl alkyl groups.

15. The composition of claim 1 wherein Z is a monocyclic aryl.

16. The composition of claim 1 wherein Z is a bicyclic aryl.

17. The composition of claim 1 wherein Z is an aromatic heterocycle.

18. The composition of claim 17, wherein said aromatic heterocycle has at least one heteroatom selected from the group consisting of nitrogen, oxygen and sulfur.

19. The composition of claim 1 wherein Z is an aryl selected from the group consisting of phenyl, napthyl, furyl, benzofuranyl, thiophenyl, benzothiophenyl, pyrrolyl, indolyl, isoindolyl, azaindolyl, pyridyl, quinolinyl, isoquinolinyl, oxazolyl, isooxazolyl, benzoxazolyl, pyrazolyl, imidazolyl, benzimidazolyl, thiazolyl, benzothiazolyl, isothiazolyl, pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl, cinnolinyl, phthalazinyl, and quinazolinyl aryls.

20. The composition of claim 1 wherein Y is a linking group —

[(CRaRb)n-A-B-C-(CRcRd)mD]p-, wherein Ra, Rb, Rc, and Rd are the same or different and at each occurrence, independently hydrogen, alkyl, or substituted alkyl;

n is 0 to 100;

m is 1 to 20;

p is 1 to 100;

A, B, C, and D are the same or different and, at each occurrence, independently absent, O, S, N, or carbonyl.