Screening assay for identification of poly(ADP-ribose) polymerase 1 inhibitors
AssigneeInstitute For Cancer Research
InventorAlexei Tulin
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Abstract
Screening methods as well as systems and kits for identifying inhibitors of DNA-independent, histone H4-dependent activation of PARP-1 are provided. The methods comprise screening molecules for their capacity to inhibit the activation and/or biologic activity of PARP-1, as measured by poly(ADP)-ribose production from nicotinamide adenine dinucleotide. PARP-1 inhibitors identified through the screening methods may be used to treat cancer in which PARP-1 activation or biologic activity plays a role.
Claims10
A method for screening a compound for capability to inhibit poly(ADP-ribose) polymerase 1 (PARP-1), comprising contacting a portion of PARP-1 comprising the catalytic domain with core histone H4 in the presence of the compound and nicotinamide adenine dinucleotide, wherein the portion is the product of caspase 3 cleavage of the full length PARP-1 protein, wherein the core histone H4 is immobilized on a plate comprising a plurality of wells;
- and determining the level of H4-dependent biologic activity of the portion of PARP-1 comprising the catalytic domain in the presence of the compound relative to the level of H4-dependent biologic activity of the portion of PARP-1 comprising the catalytic domain in the absence of the compound.
A kit for screening a compound for capability to inhibit poly(ADP-ribose) polymerase 1 (PARP-1), comprising:
- a portion of PARP-1 comprising the catalytic domain, wherein the portion is the product of caspase 3 cleavage of the full length PARP-1 protein;
- nicotinamide adenine dinucleotide;
- core histone H4 immobilized on a plate comprising a plurality of wells;
- and instructions for using the kit.
Dependencies are inferred from the claim text.