Arginine analogs, and methods for their synthesis and use

• US 7,879,979 B2
• Filed: 01/19/2006
• Issued: 02/01/2011
• Est. Priority Date: 01/21/2005
• Status: Active Grant

• Current Assignee: Alere San Diego Incorporated (Abbott Laboratories Incorporated)
• Original Assignee: Alere International
• Inventors: Buechler, Kenneth F., Dinauer, Violeta, Banaszczyk, Mariusz G., Noar, Joseph Barry
• Primary Examiner(s): Shiao; Rei-tsang
• Application Number:
  US11/795,173
• Publication Number:
  US 20080293920A1
• Time in Patent Office: 1,839 Days
• Field of Search: 530/363, 549/62, 562/561
• US Class Current: 530/363
• CPC Class Codes: C07C 323/59 with acylated amino groups bound to the carbon skeleton
• CPC Class Codes: C07C 323/60 with the carbon atom of at least one of the carboxyl groups bound to nitrogen atoms
• CPC Class Codes: C07C 327/32 having sulfur atoms of esterified thiocarboxyl groups bound to carbon atoms of hydrocarbon radicals substituted by carboxyl groups
• CPC Class Codes: C07D 333/36 Nitrogen atoms

Abstract

The present invention relates to novel arginine analogs, and methods for their synthesis and use. Such analogs are designed to provide a protected or free thiol (—SH) group, thereby providing a convenient linkage chemistry for coupling to a suitable group on a target such as a protein, polypeptide, detectable label or solid phase, and at a site distal to the guanidino group. Arginine analog conjugates are useful for generating antibodies that can bind specifically with dimethylarginine, which can be detected using such antibodies in immunoassays.

Claims

1. A compound or salt thereof, said compound having the structure:

2. A compound or salt thereof according to claim 1, wherein R¹ is H, and R² is a conjugating group having the structure

3. A compound or salt thereof according to claim 2, wherein Y is absent and Z is a thiolactone.

4. A compound or salt thereof according to claim 3, wherein said thiolactone has 5 ring members.

5. A compound or salt thereof according to claim 2, wherein Y is N(H)—

C_0-6 alkyl and Z is a thioester.

6. A compound or salt thereof according to claim 5, wherein Y has the structure

—

N(H)CH₂C(O)CH₂—

.

7. A compound or salt thereof according to claim 6, wherein R³ is selected from the group consisting of —

CH₃, —

CH₂CH₃, —

OCH₃, —

OCH₂CH₃, —

OCH₂C₆H₅, —

OCH₂CH₂C₆H₅, and —

C₆H₅.

8. A compound or salt thereof according to claim 2, wherein R¹ is H, and R² is a conjugating group having the structure:

9. The compound or salt thereof according to claim 8, wherein Z is an alkyl thiol.

10. The compound or salt thereof according to claim 9, wherein Z is —

(CH₂)_1-2SH.

11. The compound or salt thereof according to claim 1, wherein R² is —

OH, and R¹ is a conjugating group having the structure

12. The compound or salt thereof according to claim 11, wherein X is C_0-4 unsubstituted alkylene, Y is N(H), and Z is a thiolactone.

13. The compound or salt thereof according to claim 12, wherein Z has 5 ring members.

14. The compound or salt thereof according to claim 11, wherein Y is C_0-4 alkylene and Z is a thioester.

15. The compound or salt thereof according to claim 14, wherein X is absent.

16. The compound or salt thereof according to claim 15, wherein R³ is selected from the group consisting of —

CH₃, —

CH₂CH₃, —

OCH₃, —

OCH₂CH₃, —

OCH₂C₆H₅, —

OCH₂CH₂C₆H₅, and —

C₆H₅.

17. The compound or salt thereof according to claim 11, wherein X is C_0-4 unsubstituted alkylene, Y is N(H) or absent, and Z is a C_1-4 thiol-substituted alkyl.

18. The compound or salt thereof according to claim 17, wherein Z has the structure:

20. A conjugate comprising a compound according to claim 1 covalently bound through a sulfhydryl group on the compound(s) to a sulfhydryl-reactive moiety on a protein, polypeptide, detectable label, nucleic acid, or solid phase.

21. The conjugate according to claim 20, wherein said sulfhydryl-reactive moiety is selected from the group consisting of a maleimide, an alkyl halide, an aryl halide, an alpha-haloacyl, and a pyridyl disulfide.

22. The conjugate according to claim 21, wherein said sulfhydryl-reactive moiety is a maleimide, whereby said compound(s) are covalently bound to said protein, polypeptide, detectable label, nucleic acid, or solid phase to provide one of the following structures:

23. The conjugate according to claim 21, wherein said detectable label is selected from the group consisting of an enzyme, a fluorophore, biotin, avidin, streptavidin, digoxigenin, maltose, oligohistidine, 2,4-dintrobenzene, phenylarsenate, and a fluorescent latex particle.

24. The conjugate according to claim 21, wherein said protein is keyhole limpet hemocyanin or bovine serum albumin.

25. The conjugate according to claim 21, wherein said compound(s) are bound to a solid phase selected from the group consisting of a membrane, a cellulose-based paper, a polymeric particle, a latex particle, a paramagnetic particle, a glass substrate, a silicon substrate, a plastic substrate, and a multiple-well plate.

19. A compound selected from the group consisting of: