Methods for identifying chemical modulators of Ras...

Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving hydrolase

Reexamination Certificate

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C435S018000

Reexamination Certificate

active

07807400

ABSTRACT:
The present invention provides a method of identifying a compound having the ability to modulate the guanine nucleotide exchange cycle of a Ras superfamily GTPase, comprising: a) contacting the compound with a guanine nucleotide exchange factor and a GTPase and obtaining a baseline fluorescence measurement; b) contacting the guanine nucleotide exchange factor and the GTPase without the compound and obtaining a baseline fluorescence measurement; c) adding a fluorophore-conjugated GTP to the components of (a) and (b), respectively; d) obtaining fluorescence measurements of the respective components of (c) over time; e) subtracting the respective baseline fluorescence measurements of (a) and (b) from each fluorescence measurement of (d); and f) comparing the resulting fluorescence values of (e), wherein a decrease or increase in the rate of fluorescence change with the compound as compared with the rate of fluorescence change without the compound identifies a compound having the ability to modulate the guanine nucleotide exchange cycle of a Ras superfamily GTPase. Further provided are compounds of the invention and pharmaceutical compositions comprising compounds of the invention useful for the treatment of cancer and neurological disorders.

REFERENCES:
patent: 2006/0263772 (2006-11-01), Shoemaker et al.
International Search Report for PCT/US05/13444, dated Feb. 14, 2007.
Stephen et al. “The nucleocapsid protein as a target for novel anti-HIV drugs” www.currentdrugdiscovery.com, pp. 33-36, 2003.
Rojas et al. “Established and Emerging Fluorescence-Based Assays for G-Protein Function: Ras-Superfamily GTPases”Combinatorial Chemistry & High Throughput Screening6(4):409 (Jun. 2003).

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