Assays and reagents for identifying anti-fungal agents, and uses related thereto
AssigneesGPC Biotech AG, The Johns Hopkins University
InventorsVivian Berlin, David E. Levin, Yoshikazu Ohya, Veronique Damagnez, Susan E. Smith
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Abstract
The present invention relates to rapid, reliable and effective assays for screening and identifying pharmaceutically effective compounds that specifically inhibit the biological activity of fungal GTPase proteins, particularly GTPases involved in cell wall integrity, hyphael formation, and/or other cellular functions critical to pathogenesis. Another aspect of the present invention relates to novel Candida genes and gene products.
Claims40
A method for inhibiting growth of a fungal pathogen comprising
- (a) forming a reaction mixture including:
- (i) a fungal geranylgeranyl transferase (GGPTase);
- (ii) a GGPTase substrate;
- (iii) a test compound, under conditions whereunder, in the absence of the test compound, the GGPTase and the GGPTase substrate interact,
- (b) detecting interaction of the GGPTase substrate with the GGPTase,
- wherein a statistically significant decrease in the interaction of the GGPTase substrate and GGPTase in the presence of the test compound, relative to the level of interaction in the absence of the test compound, indicates a potential GGPTase inhibitory activity for the test compound;
- and
- (c) contacting the pathogen with the test compound identified as having a potential GGPTase inhibitory activity, whereby growth of the fungal pathogen is inhibited.
A method for inhibiting growth of a fungal pathogen comprising
- (i) identifying a patient having a fungal pathogen infection,
- (ii) (a) forming a reaction mixture including:
- (i) a fungal geranylgeranyl transferase (GGPTase);
- (ii) a GGPTase substrate;
- (iii) a test compound, under conditions whereunder, in the absence of the test compound, the GGPTase and the GGPTase substrate interact, and
- (b) detecting interaction of the GGPTase substrate with the GGPTase,
- wherein a statistically significant decrease in the interaction of the GGPTase substrate and GGPTase in the presence of the test compound, relative to the level of interaction in the absence of the test compound, indicates a potential GGPTase inhibitory activity for the test compound, and
- (iii) administering to the patient the test compound identified as having a potential GGPTase inhibitory activity, whereby growth of the fungal pathogen is inhibited.
A method for inhibiting growth of a fungal pathogen comprising
- (a) forming a cell-free reaction mixture including:
- (i) a fungal geranylgeranyl transferase (GGPTase);
- (ii) a peptidyl substrate for the GGPTase;
- (iii) an activated geranylgeranyl group;
- (iv) a divalent cation;
- and
- (v) a test compound,
- (b) detecting prenylation of the peptidyl substrate for the GGPTase in the reaction mixture,
- wherein a statistically significant decrease in the prenylation of the peptidyl substrate in the presence of the test compound, relative to the level of prenylation in the absence of the test compound, indicates a potential anti-fungal activity for the test compound, and
- (c) administering an effective amount of the test compound identified as having a potential GGPTase inhibitory activity to a patient having a fungal pathogen whereby growth of the fungal pathogen is inhibited.
Dependencies are inferred from the claim text.