Method of identifying inhibitors of Tie-2
View Patent ↗The present invention relates to polypeptides which comprise the ligand binding domain of Tie-2, crystalline forms of these polypeptides and the use of these crystalline forms to determine the three dimensional structure of the catalytic domain of Tie-2. The invention also relates to the use of the three dimensional structure of the Tie-2 catalytic domain both alone, or in complex with inhibitors, in methods of designing and/or identifying potential inhibitors of Tie-2 activity, for example, compounds which inhibit the binding of a native substrate to the Tie-2 catalytic domain.
1. A method of identifying a compound which is a potential inhibitor of a Tie-2 protein, said method comprising the steps of:
(a) crystallizing a crystal of a polypeptide consisting of the catalytic domain of a Tie-2 protein as defined in FIG. 2 ;
(b) obtaining the atomic coordinates of the crystal of the polypeptide;
(c) using said atomic coordinates to define the active subsites of Tie-2; and
(d) identifying a compound which binds to one or more active subsites;
wherein the compound which binds to the active subsite or sites is a potential inhibitor of a Tie-2 protein.
2. The method of claim 1 , further comprising the step of:
(d) assessing the ability of the compound identified in step (c) to inhibit Tie-2.
3. The method of claim 1 , wherein the Tie-2 protein is a mammalian protein.
4. The method of claim 2 , wherein the Tie-2 protein is a human protein.
5. The method of claim 4 , wherein the Tie-2 protein is wild type human Tie-2.
6. The method of claim 1 , wherein said crystal further comprises a ligand bound to said catalytic domain.
7. The method of claim 4 , wherein the polypeptide comprises amino acids 802-1124 of SEQ ID NO: 1.
8. The method of claim 4 , wherein the compound is of the formula:
9. The method of claim 8 , wherein the crystal has unit cell parameters wherein a is about 96 Å, b is about 118 Å, c is about 78 Å and α=β=γ=90°.
10. The method of claim 4 , wherein the compound is of the formula:
11. The method of claim 10 , wherein the crystal has unit cell parameters wherein a and b are about 86.0 Å, c is about 112.0 Å and α=β=γ=90°.
12. The method of claim 4 , wherein the compound is of the formula:
13. The method of claim 12 , wherein the crystal has unit cell parameters wherein a and b are about 86.0 Å, and c is about 112 Å and α=β=γ=90°.
14. The method of claim 4 , wherein the compound is of the formula:
15. The method of claim 14 , wherein the crystal has unit cell parameters wherein a and b are about 86.0 Å, and c is about 112 Å and α=β=γ=90°.