Neuroprotective chemicals and methods for identifying and using same
Provided herein are methods for identifying a compound having cell-protective (e.g., neuroprotective) activity. Compounds identified therefrom are also provided. These compounds can be used to treat various diseases, disorders, or conditions associated with, for example, unwanted cell death.
1. A method for identifying a compound having cell-protective activity, comprising:
exposing a population of cells to a test compound and a DNA-damaging agent;
determining survival rate of the population of cells;
wherein an increase in the survival rate compared to a control that is not treated with the test compound is indicative of cell-protective activity of the test compound; and
incubating the test compound with nicotinamide phosphoribosyltransferase (NAMPT) and measuring an activity of the NAMPT, wherein an increase in the activity of the NAMPT additionally confirms the cell-protective activity of the test compound.
2. The method of claim 1 , wherein the exposing step comprises incubating the population of cells with the test compound and the DNA-damaging agent.
3. The method of claim 1 , wherein the exposing step comprises incubating the population of cells with the test compound first, followed by the DNA-damaging agent.
4. The method of claim 1 , wherein the exposing step comprises incubating the population of cells with the DNA-damaging agent first, followed by the test compound.
5. The method of claim 1 , wherein the DNA-damaging agent activates poly-ADP-ribose polymerase (PARP).
6. The method of claim 5 , wherein the DNA-damaging agent leads to decline in nicotinamide adenine dinucleotide (NAD) level.
7. The method of claim 1 , wherein the DNA-damaging agent is an anthracycline.
8. The method of claim 7 , wherein the anthracycline is one or more of daunorubicin, doxorubicin, epirubicin, idarubicin, valrubicin and mitoxantrone.
9. The method of claim 8 , wherein the anthracycline is doxorubicin.
10. The method of claim 1 , wherein the cells are mammalian cells.
11. The method of claim 1 , wherein the test compound is provided at about 1-20 uM, or about 2-10 uM or about 5 uM.
12. The method of claim 1 , wherein the test compound is a small molecule.
13. The method of claim 1 , wherein the DNA-damaging agent is provided at about 0.01-20 uM, or about 0.05-10 uM, or about 0.1-1.5 uM, or about 0.5 uM.
14. The method of claim 1 , further comprising determining the test compound having said cell-protective activity protects the cells from toxicity mediated by the DNA-damaging agent.
15. The method of claim 1 , further comprising determining the test compound having said cell-protective activity does not protect the cells from toxicity induced by a toxin selected from the group consisting of bortezomib, staurosporine, taxol, brefeldin, cytochalasin D, TNFa and TRAIL.
16. The method of claim 12 , further comprising determining the test compound having said cell-protective activity increases nicotinamide adenine dinucleotide (NAD) level.
17. The method of claim 1 , further comprising determining the test compound having said cell-protective activity binds and/or activates the NAMPT.
18. The method of claim 1 , wherein the cell-protective activity is neuroprotective activity which includes one or more of: promoting survival, health, integrity, growth, development and/or function of neurons, protecting neurons from cell death, apoptosis and/or degeneration, and stimulating neurogenesis.