Method and compounds for inhibition of cell death
The invention is directed to methods and compositions for inhibiting caspase-independent apoptosis. In particular, methods and compositions for inhibiting Omi/HtrA2 activity, as well as method for identifying other inhibitors of Omi/HtrA2. Also disclosed are Omi/HtrA2 specific substrates and methods for identifying other substrates of Omi/HtrA2.
1. A method of inhibiting Omi/HtrA2 activity, from about 20% to about 90% comprising:
contacting a cell having Omi/HtrA2 activity with an apoptosis inhibiting compound; and
monitoring the inhibition of Omi/HtrA2 activity, wherein the apoptosis inhibiting compound is selected from the group consisting of:
where R 1 is selected from the group consisting of a chloride, nitro group, a carboxy group, a hydroxide, an aliphatic group, an aromatic group, an acyl group, an alkoxy group, an alkylene group, an alkenylene group, an alkynylene group, a hydroxycarbonylalkyl group, an anhydride, an amide, an amine, and a heterocyclic aromatic group, and where R 2 is a methoxy group.
2. The method of claim 1 , wherein the step of contacting the cell comprises contacting the cell in vivo.
3. The method of claim 1 , wherein the step of contacting the cell comprises contacting the cell in vitro.
4. The method of claim 1 , wherein the apoptosis inhibiting compound is formula I, wherein R 1 is a nitro group (ucf-101).
5. The method of claim 1 , wherein the apoptosis inhibiting compound is formula II, wherein R 1 is a chloride and R 2 is a methoxy group (ucf-102).
6. The method of claim 1 , wherein the apoptosis inhibiting compound is formula III, wherein R 1 is CO 2 H (ucf-103).
7. The method of claim 1 , wherein the apoptosis inhibiting compound is formula IV, wherein R 1 is a nitro group (ucf-104).
8. The method of claim 1 , wherein the step of monitoring Omi/HtrA2 activity comprises monitoring a change in fluorescene of an Omi/HtrA2 substrate coupled to a fluorescent marker.
9. The method of claim 1 , wherein the step of monitoring the inhibition of Omi/HtrA2 activity further comprises monitoring apoptosis of the cell.
10. The method of claim 8 , wherein the fluorescent marker is selected from the group consisting of fluorescein isothiocyanate (FITC), cyanine dye-5 (CYS), cyanine dye-3 (Cy3), cyanine dye-7 (Cy7), allophycocyanin (APC), tetramethyl rhodamine isothiocyanate (TRITC), and phycoerythrin (PE).
11. A method of inhibiting Omi/HtrA2 activity, comprising:
contacting a cell having Omi/HtrA2 activity with an apoptosis inhibiting compound; and
monitoring the inhibition of Omi/HtrA2 activity,
wherein the apoptosis inhibiting compound is selected from the group consisting of Ucf-101 Ucf-102, Ucf-103 and Ucf-104.
12. A method for inhibiting caspase-independent apoptosis in a cell comprising:
contacting a cell having Omi/HtrA2 activity with at least one apoptosis inhibiting compound selected from the group consisting of Ucf-101 Ucf-102, Ucf-103 and Ucf-104, such that the apoptosis inhibiting compound interacts with Omi/HtrA2 to inhibit the activity of Omi/HtrA2, wherein the inhibition of Omi/HtrA2 activity reduces apoptosis in the cell; and monitoring the inhibition of apoptosis.