Cyclopropene amino acids and methods

Patent metadata

Field	Value
Publication number	US-10774039-B2
Application number	US-201515124632-A
Country	US
Kind code	B2
Filing date	Mar 10, 2015
Priority date	Mar 14, 2014
Publication date	Sep 15, 2020
Grant date	Sep 15, 2020

Abstract

The invention relates to a polypeptide comprising an amino acid having a cyclopropene group wherein said cyclopropene group is joined to the amino acid via a carbamate group. Suitably the cyclopropene group is a 1,3-disubstituted cyclopropene such as a 1,3-dimethylcyclopropene. Suitably the cyclopropene group is present as a residue of a lysine amino acid. The invention also relates to methods of making the polypeptides. The invention also relates to an amino acid comprising cyclopropene wherein said cyclopropene group is joined to the amino acid moiety via a carbamate group.

First claim

What is claimed is:

1. A polypeptide comprising an amino acid having a cyclopropene group wherein said cyclopropene group is linked to the amino acid via a carbamate group and the carbamate group does not form a part of the polypeptide backbone, wherein said cyclopropene group is a 1,3-disubstituted cyclopropene.

2. A polypeptide according to claim 1 wherein said cyclopropene is a 1,3-dimethylcyclopropene.

3. A polypeptide according to claim 1 wherein said cyclopropene group is present as a residue of a lysine amino acid.

4. A polypeptide according to claim 1 further comprising a tetrazine compound linked to said cyclopropene group.

5. An amino acid comprising cyclopropene wherein said cyclopropene group is linked to the amino acid moiety via a carbamate group and the carbamate group does not form a part of the peptide backbone, wherein said cyclopropene group is a 1,3-disubstituted cyclopropene.

6. An amino acid according to claim 5 wherein said cyclopropene is a 1,3-dimethylcyclopropene.

7. An amino acid according to claim 5 wherein said amino acid is a lysine amino acid.

8. An amino acid according to claim 7 which comprises N ε -[((2-methylcycloprop-2-en-1-yl)methoxy)carbonyl]-1-lysine.

9. An amino acid according to claim 7 which consists of

10. A method of producing a polypeptide comprising a cyclopropene group wherein said cyclopropene group is joined to an amino acid moiety of the polypeptide via a carbamate group, said method comprising genetically incorporating said amino acid comprising said cyclopropene group joined to said amino acid moiety via said carbamate group, into said polypeptide.

11. A method according to claim 10 wherein producing the polypeptide comprises
(i) providing a nucleic acid encoding the polypeptide which nucleic acid comprises an orthogonal codon encoding the amino acid having a cyclopropene group;
(ii) translating said nucleic acid in the presence of an orthogonal tRNA synthetase/tRNA pair capable of recognising said orthogonal codon and incorporating said amino acid having a cyclopropene group into the polypeptide chain.

12. A method according to claim 10 wherein said orthogonal codon comprises an amber codon (TAG), said tRNA comprises MbtRNA CUA and said tRNA synthetase comprises MbPylRS; or wherein said orthogonal codon comprises an amber codon (TAG), said tRNA comprises MmtRNA CUA and said tRNA synthetase comprises MmPylRS.

13. A method according to claim 10 wherein said carbamate group does not form a part of the peptide backbone, and wherein said cyclopropene group is a 1,3-disubstituted cyclopropene.

14. A method of producing a polypeptide comprising a tetrazine group, said method comprising providing a polypeptide according to claim 1 , contacting said polypeptide with a tetrazine compound, and incubating to allow joining of the tetrazine to a cyclopropene group of the polypeptide by an inverse electron demand Diels-Alder cycloaddition reaction.

15. A method according to claim 14 wherein said reaction is allowed to proceed for 10 minutes or less, preferably for 1 minute or less, preferably for 30 seconds or less.

16. A polypeptide according to claim 1 wherein said polypeptide comprises two or more amino acids each having a cyclopropene group, wherein each said cyclopropene group is linked to each said amino acid via a carbamate group and wherein each carbamate group does not form a part of the polypeptide backbone.

17. A polypeptide according to claim 16 wherein said polypeptide comprises four amino acids each having a cyclopropene group.

18. An antibody drug conjugate (ADC) comprising a polypeptide according to claim 1 .

Assignees

• Res & Innovation Uk

Inventors

• Elliott Thomas

Classifications

• C12Y601/01

  Ligases forming aminoacyl-tRNA and related compounds (6.1.1) · CPC title

• C12P21/02

  having a known sequence of two or more amino acids, e.g. glutathione · CPC title

• C12N9/2462

  Lysozyme (3.2.1.17) · CPC title

• C07K14/43595 Primary

  from coelenteratae, e.g. medusae · CPC title

• C07K1/1072 Primary

  by covalent attachment of residues or functional groups · CPC title