Compositions and methods for inducing conformational changes in cereblon and other E3 ubiquitin ligases

Patent metadata

Field	Value
Publication number	US-10092555-B2
Application number	US-201514752588-A
Country	US
Kind code	B2
Filing date	Jun 26, 2015
Priority date	Jun 27, 2014
Publication date	Oct 9, 2018
Grant date	Oct 9, 2018

Abstract

Provided herein are compositions, therapeutic methods, screening methods, computational methods and biomarkers based upon the elucidation of the interaction among cereblon, its substrates and certain compounds or agents, including small molecules, peptides, and proteins.

First claim

What is claimed is:

1. A method of identifying a test compound that induces a cereblon protein (CRBN) conformational change or an alteration of properties of a CRBN surface, wherein the method comprises:
(a) (i) obtaining a first crystal of CRBN and a reference compound or an apo-CRBN crystal, and (ii) determining the three-dimensional structure of the first crystal by x-ray diffraction to obtain a first set of atomic coordinates;
(b) (i) obtaining a second crystal comprising CRBN and the test compound, and (ii) determining the three-dimensional structure of the second crystal by x-ray diffraction to obtain a second set of atomic coordinates; and
(c) employing on a computer, the structural coordinates from the first set of atomic coordinates and the second set of atomic coordinates and comparing the two sets with one another wherein identification of a test compound that induces a conformational change or alteration of properties of a CRBN surface is made by identifying a shift in the protein backbone or sidechains in the second set of atomic coordinates as compared to the first set atomic coordinates;
wherein the CRBN conformational change or the alteration of properties of the CRBN surface occurs in a CRBN modifying agent (CMA) binding pocket of the CRBN, or within an adjacent region thereof, and
wherein said first and second crystals comprising CRBN form in space group P2 1 2 1 2 1 or / 23 and are human or murine CRBN.

2. The method of claim 1 , wherein the method further comprises assaying the identified test compound's biological activity.

3. The method of claim 1 , wherein the conformational change or alteration has an effect on W380 W386 and/or W400 of CRBN; has an effect on E377 of CRBN, or has an effect on V388 of CRBN; wherein the CRBN is human CRBN and the numbering corresponds to native human CRBN.

4. The method of claim 1 wherein the CRBN and reference compound of (a) is further bound to Damaged DNA Binding protein 1 (DDB1), Cullin-4 (Cul4), homeobox-leucine zipper protein ROC1 (Roc 1 ), or any combination thereof.

5. The method of claim 1 , wherein the CRBN and reference compound of (a) is further bound to DDB1.

6. The method of claim 1 , wherein the CRBN of (b) is further bound to Damaged DNA Binding protein 1 (DDB1), Cullin-4 (Cul4), homeobox-leucine zipper protein ROC1 (Roc 1 ), or any combination thereof.

7. The method of claim 1 , wherein the CRBN of (b) is further bound to DDB1.

Assignees

• Celgene Corp

Inventors

• Chamberlain Philip
• Cathers Brian E
• Lopez-Girona Antonia

Classifications

• A61P35/00

  Antineoplastic agents · CPC title

• A61P43/00

  Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00 · CPC title

• C12Q1/37

  involving peptidase or proteinase · CPC title

• G16B15/00

  ICT specially adapted for analysing two-dimensional [2D] or three-dimensional [3D] molecular structures, e.g. structural or functional relations or structure alignment · CPC title

• A61K31/454 Primary

  containing a five-membered ring with nitrogen as a ring hetero atom, e.g. pimozide, domperidone · CPC title