Functionally modified polypeptides and radiobiosynthesis
AssigneeIkaria, Inc.
InventorStephen DiMagno
Life of this patent
Estimating the term…
Abstract
Provided herein are compositions and methods for generating polypeptides using non-natural amino acids (nnAAs) and genetic machinery, wherein the modified polypeptides, such as therapeutic polypeptides, bind to albumin, such as serum albumin. Methods of substituting a non-natural amino acid in a first polypeptide to obtain a modified polypeptide, the nnAA in some instances comprising an albumin targeting group, are disclosed, as are methods for making populations of such modified polypeptides. A therapeutic polypeptide, interleukin-1 receptor antagonist (IL-1RA) is exemplified using the disclosed methods.
Claims24
A method of substituting a natural amino acid in a first polypeptide with a non-natural amino acid to obtain a modified polypeptide, wherein the non-natural amino acid comprises a moiety selected from the group consisting of:
- an albumin-targeting group, a radiolabel, and a radiolabelled albumin-targeting group, comprising:
- (a) selecting at least one amino acid residue in the first polypeptide to be substituted with the non-natural amino acid;
- (b) selecting a polynucleotide encoding the first polypeptide;
- (c) modifying the polynucleotide such that the amino acid residue to be substituted is encoded by a nonsense codon;
- (d) expressing the modified polynucleotide in a cell, wherein the cell is in the presence of the non-natural amino acid and expresses a suppressor tRNA and its cognate tRNA synthetase wherein the suppressor tRNA recognizes the nonsense codon of step (c), and the cell incorporates the non-natural amino acid into the modified polypeptide at the nonsense codon of step (c) during translation.
10-33. (canceled)
A cell comprising a modified polynucleotide encoding a modified polypeptide, a suppressor tRNA, and a cognate tRNA synthetase,
- wherein the modified polynucleotide comprises one or more nonsense codons recognized by the suppressor tRNA, and the cognate tRNA synthetase,
- wherein the modified polypeptide comprises one or more non-natural amino acids at positions encoded by the nonsense codons, and
- wherein the non-natural amino acids comprise a moiety selected from the group consisting of:
- an albumin-targeting group, a radiolabel, and a radiolabelled albumin-targeting group.
35-37. (canceled)
The cell of claim 37, wherein the non-natural amino acid is Nε-(4-(4-iodophenyl)butanoyl)lysine, or a pharmaceutically acceptable salt thereof.
The cell of claim 37, wherein the non-natural amino acid comprises a radiolabel.
(canceled)
42-123. (canceled)
A composition comprising a plurality of identical polypeptides,
- wherein each polypeptide comprises one or more non-natural amino acids, and
- wherein the non-natural amino acids comprise a moiety selected from the group consisting of:
- an albumin-targeting group, a radiolabel, and a radiolabelled albumin-targeting group.
A non-naturally occurring amino acid selected from the group consisting of:
A non-naturally occurring amino acid selected from the group consisting of
- Nε-(4-(4-iodophenyl)butanoyl)lysine;
- Nε-(4-(4-iodophenyl)butanethioyl)lysine, and
- Nε-(3-(4-iodophenyl)propanoyl)lysine.
Dependencies are inferred from the claim text.