# US 2024/0199686 A1 · C-Terminal Peptide Modification

> Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt

- Number: US 2024/0199686 A1
- Document ID: US20240199686A1
- Status: Published application
- Kind: A1
- Application number: 18/281,787
- Filed: 2022-03-18
- Published: 2024-06-20
- Earliest claimed: 2021-03-18
- Applicants: Technische Universiteit Delft
- Inventors: Rienk Eelkema; Peggy Bohlaender
- Web page: https://patentgrep.com/patent/US20240199686A1

## CPC

- C07K 1/1075: {by covalent attachment of amino acids or peptide residues}
- C07K 1/1077: {by covalent attachment of residues other than amino acids or peptide residues, e.g. sugars, polyols, fatty acids}
- A61K 47/6801: {Drug-antibody or immunoglobulin conjugates defined by the pharmacologically or therapeutically active agent}
- C07K 1/10: using coupling agents {(C07K1/006 takes precedence)}
- C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00)
- A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D)

## Abstract

The invention provides a method for providing a cargo to a C-terminal end of a peptide, the method comprising a first stage and a second stage, wherein the first stage comprises reacting the C-terminal end of the peptide with a first reactant in the presence of a first catalyst and first radiation to provide a first intermediate, wherein the first catalyst is configured to decarboxylate the C-terminal end of the peptide in the presence of the first radiation; and wherein the second stage comprises exposing the first intermediate to a second reactant.

## Claims

### Claim 1

A method for providing a cargo to a C-terminal end of a peptide, the method comprising a first stage and a second stage, wherein the first stage comprises reacting the C-terminal end of the peptide with a first reactant in the presence of a first catalyst and first radiation to provide a first intermediate, wherein the first catalyst is configured to decarboxylate the C-terminal end of the peptide in the presence of the first radiation, and wherein the first reactant has a first chemical structure according to formula I:
wherein R and R′are each independently selected from the group consisting of H, and alkyl groups;
wherein R″ is an electron withdrawing group comprising a functional group selected from the group consisting of an ester, a thioester, an amide, a ketone, a nitro, a sulfoxide, a sulfone, a phosphate ester, an acylhydrazide, a cyano group, and a trihalogenmethyl group;
wherein n1 and n2 are each independently selected from the range of 1-2;
and wherein the second stage comprises exposing the first intermediate to a second reactant, wherein the second reactant has a second chemical structure according to formula II:
wherein X is O or NH, and wherein R1 comprises the cargo.

### Claim 2 (depends on 1)

The method according to claim 1, wherein n1=1, and wherein n2=1.

### Claim 3 (depends on 1)

The method according to claim 1, wherein R and R′ are selected from the group consisting of H and alkyl groups comprising 1-6 C atoms.

### Claim 4 (depends on 3)

The method according to claim 3, wherein one or more of R and R′ are H.

### Claim 5 (depends on 1)

The method according to claim 1, wherein R″ comprises a functional group selected from the group consisting an ester, a cyano group, or —CF3.

### Claim 6 (depends on 2)

The method according to claim 2, wherein the first reactant has a first chemical structure according to structure III:

### Claim 7 (depends on 1)

The method according to claim 1, wherein the first catalyst comprises riboflavin tetrabutyrate.

### Claim 8 (depends on 1)

The method according to any one of the preceding claim 1, wherein the first radiation is selected from the range of 375-525 nm.

### Claim 9 (depends on 1)

The method according to claim 1, wherein the first stage is executed in a first mixture comprising a first solvent, wherein the first solvent comprises one or more of DMF, DMSO, and water.

### Claim 10 (depends on 9)

The method according to claim 9, wherein the first mixture comprises ≤1 ppm dissolved oxygen.

### Claim 11 (depends on 1)

The method according to claim 1, wherein the method further comprises an intermediate stage, wherein the intermediate stage comprises separating the first intermediate from the first reactant.

### Claim 12 (depends on 1)

The method according to claim 1, wherein the second stage is performed in a degassed buffer under inert gas.

### Claim 13 (depends on 1)

The method according to claim 1, wherein the peptide comprises a C-terminal residue selected from the group comprising alanine, arginine, asparagine, aspartate, cysteine, glutamate, glutamine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, pyrrolysine, selenocysteine, serine, threonine, tryptophan, tyrosine, and valine.

### Claim 14 (depends on 1)

The method according to claim 1, wherein the cargo comprises a second peptide, wherein the second peptide has an N-terminal cysteine residue.

### Claim 15 (depends on 1)

The method according to claim 1, wherein the cargo comprises an antibody.

### Claim 16 (depends on 1)

A peptide-cargo conjugate obtainable using the method of claim 1, wherein the peptide-cargo conjugate has a chemical structure according to formula IV:
wherein R and R′ are each independently selected from the group consisting of H, and alkyl groups, wherein R3 is the peptide, wherein R″ is an electron withdrawing group comprising a functional group selected from the group consisting of an ester, a thioester, an amide, a ketone, a nitro, a sulfoxide, a sulfone, a phosphate ester, an acylhydrazide, a cyano group, and a trihalogenmethyl group, wherein n1 is 1, wherein X is O or NH, and wherein R1 comprises the cargo.

### Claim 17 (depends on 16)

The peptide-cargo conjugate according to claim 16 for use as an antibody drug conjugate.

### Claim 18 (depends on 16)

A peptide array comprising the peptide-cargo conjugate according to claim 16.

### Claim 19 (depends on 16)

Use of the peptide-cargo conjugate according to claim 16 for peptide sequencing.