Functionalized 1,2,4,5-tetrazine compounds for use in bioorthogonal coupling reactions

• US 10,611,738 B2
• Filed: 02/26/2018
• Issued: 04/07/2020
• Est. Priority Date: 10/24/2012
• Status: Active Grant

• Current Assignee: The General Hospital Corporation (Mass General Brigham, Inc.)
• Original Assignee: The General Hospital Corporation (Mass General Brigham, Inc.)
• Inventors: Hilderbrand, Scott A., Devaraj, Neal K., Weissleder, Ralph, Karver, Mark R.
• Primary Examiner(s): Balasubramanian, Venkataraman
• Application Number:
  US15/904,927
• Publication Number:
  US 20190040021A1
• Time in Patent Office: 771 Days
• Field of Search: 544179
• CPC Class Codes: C07D 257/08 Six-membered rings
• CPC Class Codes: C07D 401/04 directly linked by a ring-member-to-ring-member bond
• CPC Class Codes: C07D 403/12 linked by a chain containing hetero atoms as chain links

Abstract

The present application relates to functionalized 1,2,4,5-tetrazine compounds. The compounds are useful in compositions and methods using bioorthogonal inverse electron demand Diels-Alder cycloaddition reactions for the rapid and specific covalent delivery of a “payload” to a ligand bound to a biological target.

Claims

1. A peptide, polypeptide or protein comprising incorporated therein a tetrazine-substituted amino acid unit derived from an amino acid of the following formula (I)

2. The peptide, polypeptide or protein of claim 1, wherein R¹ is hydrogen, (C₁-C₆)alkyl, or (C₁-C₆)haloalkyl.

3. The peptide, polypeptide or protein of claim 1, wherein A is according to formula (A2):

4. The peptide, polypeptide or protein of claim 3, wherein n1 is 3, 4 or 5, and n2 is 0, 1, or 2.

5. The peptide, polypeptide or protein of claim 1, wherein A is according to formula (A3):

6. The peptide, polypeptide or protein of claim 5, wherein n2 is other than 0 and n1 is 4.

7. The peptide, polypeptide or protein of claim 1, wherein A is according to formula (A4):

8. The peptide, polypeptide or protein of claim 7, wherein n1 is 1 or 2 and n2 is 5.

9. The peptide, polypeptide or protein of claim 1, wherein A is according to formula (A5):

10. The peptide, polypeptide or protein of claim 9, wherein n1 is 3, 4 or 5 and n2 is 0.

11. The peptide, polypeptide or protein of claim 1, wherein the amino acid is selected from amino acids of the following formulae:

12. The peptide, polypeptide or protein of claim 1, wherein the amino acid has (L) configuration.

13. A method of preparing a peptide, polypeptide or protein comprising incorporated therein a tetrazine-substituted amino acid unit comprising reacting a peptide, polypeptide or protein under peptide coupling conditions with a compound according the following formula (I-1):

14. The method of claim 13, wherein R^N is hydrogen, tert-butyloxycarbonyl or 9-fluorenylmethoxycarbonyl and R^C is hydrogen or (C₁-C₆)alkyl.