Structure-based hepatitis C virus drug design
View Patent ↗The extracellular loop of CD81 is a cellular receptor for the E2 protein of hepatitis C virus. A CD81 crystal structure has been elucidated and is provided for use in the structure-based design of compounds which bind to CD81 and thus block the binding of HCV. Methods such as docking and de novodrug design can be used.
1. A method for identifying a ligand for the large extracellular loop of CD81, comprising the steps of (a) providing a 3D structural representation of the large extracellular loop of CD81, wherein the 3D structural representation comprises a D helix having a residue Phe186, and wherein the 3D structural representation of the large extracellular loop of CD81 is set forth in the atomic co-ordinates of Table 1, or variants of the atomic co-ordinates of Table 1 thereof, in which the r.m.s. deviation of the x, y and z co-ordinates for all heavy atoms is less than 2.5 Å; (b) providing a 3D structural representation of a potential ligand; (c) using a computer to dock the 3D structural representation of the potential ligand with the 3D structural representation of the large extracellular loop of CD81 at a site comprising the residue Phe186, wherein a potential ligand that docks with the large extracellular loop of CD81 at the site comprising the residue Phe186 is identified as a ligand for the large extracellular loop of CD81; (d) contacting the ligand identified in step (c) with CD81 or a fragment thereof containing the large extracellular loop, and (e) assaying the interaction between the ligand and CD81 or the fragment thereof containing the large extracellular loop to determine whether the ligand identified in step (c) is a ligand for the large extracellular loop of CD81.
2. The method of claim 1 , wherein using the computer to dock the 3D structural representations of the potential ligand with the 3D structural representation of the large extracellular loop of CD81 comprises geometric matching or minimising the energy of interaction between the ligand and the large extracellular loop of CD81.
3. The method of claim 2 , wherein the step of using the computer to dock the potential ligand with the large extracellular loop of CD81 comprises screening members of a structural library of potential ligands to identify a ligand for CD81.
4. The method of claim 1 , further comprising comparing the structures of the ligand identified in step (c), thereby defining a pharmacophore.
5. The method of claim 4 , wherein the pharmacophore is used in de novo drug design.
6. The method of claim 1 , wherein the 3D structural representation of the large extracellular loop of CD81 includes van der Waals contacts, electrostatic interactions, and/or hydrogen bonding opportunities on the CD81 surface.
7. The method of claim 1 , wherein the site comprising the residue Phe186 further comprises one or more of Leu154, Thr163, Ile181, Ile182, Leu185, Glu188, and Asp196.
8. The method of claim 1 , wherein the site comprising the residue Phe186 is the D-helix region Asn180-Phe186.
9. A method for identifying a ligand for the large extracellular loop of CD81, comprising:
(a) contacting a ligand with CD81 or a fragment thereof containing the large extracellular loop, and
(b) assaying the interaction between the ligand and CD81 or the fragment thereof containing the large extracellular loop to determine whether the ligand is a ligand for CD81,
wherein the ligand of step (a) was identified by a method comprising:
providing a 3D structural representation of the large extracellular loop of CD81, wherein the 3D structural representation comprises a D helix having a residue Phe186, and wherein the 3D structural representation of the large extracellular loop of CD81 is set forth in the atomic co-ordinates of Table 1, or variants of the atomic co-ordinates of Table 1, in which the r.m.s. deviation of the x, y and z co-ordinates for all heavy atoms is less than 2.5 Å;
providing a 3D structural representation of a potential ligand;
using a computer to dock the 3D structural representation of the potential ligand with the 3D structural representation of the large extracellular loop of CD81 at a site comprising the residue Phe186, wherein a potential ligand that docks with the large extracellular loop of CD81 at a site comprising the residue Phe186 is identified as a ligand for the large extracellular loop of CD81.
10. The method of claim 9 , wherein the site comprising the residue Phe186 further comprises one or more of Leu154, Thr163, Ile181, Ile182, Leu185, Glu188, and Asp196.
11. The method of claim 9 , wherein the site comprising the residue Phe186 is the D-helix region Asn180-Phe186.