Photocaged citrulline analogs and methods for site-specific incorporation of citrulline into proteins

Patent metadata

Field	Value
Publication number	US-2024166594-A1
Application number	US-202218279931-A
Country	US
Kind code	A1
Filing date	Mar 11, 2022
Priority date	Mar 16, 2021
Publication date	May 23, 2024
Grant date	—

Abstract

The invention provides novel compounds and methods for site-specific incorporation of citrulline into peptides and proteins in mammalian cells. The invention also relates to modified peptides and proteins with site specific citrullination and pharmaceutical compositions and methods of preparation and use thereof.

First claim

1 . A compound having the structural formula (I):

wherein R is a photoreleasable group.

2 . The compound of claim 1 , wherein R is:

wherein each of R a and R b is selected from H or OR c , wherein R c is a C 1 -C 6 alkyl group.

3 . The compound of claim 2 , wherein R is:

4 . The compound of claim 1 , wherein R is:

5 . The compound of claim 1 , wherein R is:

6 . The compound of claim 1 , wherein R is:

7 . A peptide comprising the compound of claim 1 .

8 . A method for treating a disease comprising administering to a patient in need thereof the peptide of claim 7 .

9 . A pharmaceutical composition comprising a peptide of claim 7 and a pharmaceutically acceptable excipient, carrier or diluent.

10 . A method for treating a disease comprising administering to a patient in need thereof the pharmaceutical composition of claim 9 .

11 . A method for site-specific citrullination in a synthetic peptide, comprising:
incorporating an unnatural citrulline analog of claim 1 at one or more positions in the synthetic peptide where citrullination is desired; and photochemically converting the unnatural citrulline analog to citrulline.

12 . The method of claim 11 , wherein incorporating an unnatural citrulline analog is achieved by using an E. coli -derived engineered leucyl-tRNA synthetase (EcLeuRS)/tRNA CUA EcLeu pair.

13 . The method of claim 12 , wherein photochemical conversion of the unnatural citrulline analog to citrulline comprises directing a UV irradiation at the synthetic peptide.

14 . The method of claim 13 , wherein the UV-Visible irradiation comprises wavelengths in the range of about 365 nm to about 700 nm.

15 . The method of claim 14 , wherein the UV irradiation comprises the wavelength of 365 nm.

16 . The method of claim 11 , wherein the photochemical conversion results in greater than 95% conversion of the unnatural citrulline analog to citrulline.

Assignees

• Univ Massachusetts

Inventors

• Thompson Paul R
• Mondal Santanu

Classifications

• C07C275/24 Primary

  containing six-membered aromatic rings · CPC title

• C07D311/18

  substituted otherwise than in position 3 or 7 · CPC title

• C07F5/022

  without C-boron linkages · CPC title

• C07K1/1075

  by covalent attachment of amino acids or peptide residues · CPC title

• C07K2/00

  Peptides of undefined number of amino acids; Derivatives thereof · CPC title