Claims
- 1. A method of identifying an antifungal compound comprising the steps of:(a) contacting fungal cells with a test compound and with a membrane potential indicator dye, and (b) detecting an increasing accumulation of said dye and retention of dye despite reduction of fungal cell viability wherein the fungal cells are selected from the group consisting of Candida (including C. albicans, C. tropicalis, C. parapsilosis, C. stellatoidea, C. krusei, C. parakrusei, C. lusitanae, C. pseudotropicalis, C. guilliermondi and C. glabrata), Aspergillus (including A. fumigatus, A. flavus, A. niger, A. nidulans, A. terreus, A. sydowi, A. flavatus, and A. glaucus), Cryptococcus, Histoplasma, Coccidioides, Paracoccidioides, Blastomyces, Basidiobolus, Conidiobolus, Rhizopus, Rhizomucor, Mucor, Absidia, Mortierella, Cunninghamella, Saksenaea, Pseudallescheria, Sporotrichosis, Fusarium, Trichophyton, Trichosporon, Microsporum, Epidermophyton, Scytalidium, Malassezia, Actinomycetes, Sporothrix, Penicillium, Saccharomyces and Pneumocystis.
- 2. A method of identifying an antibacterial compound comprising the steps of:(a) contacting bacterial cells with a test compound and with a membrane potential indicator dye, and (b) detecting an increasing accumulation of said dye and retention of dye despite reduction of bacterial cell viability wherein the bacterial cells are gram-negative bacteria selected from the group consisting of Acidaminococcus, Acinetobacter, Aeromonas, Alcaligenes, Bacteroides, Bordetella, Branhamella, Brucella, Calymmatobacterium, Campylobacter, Cardiobacterium, Chromobacterium, Citrobacter, Edwardsiella, Enterobacter, Escherichia, Flavobacterium, Francisella, Fusobacterium, Haemophilus, Klebsiella, Legionella, Moraxella, Morganella, Neisseria, Pasturella, Plesiomonas, Proteus, Providencia, Pseudomonas, Salmonella, Serratia, Shigella, Streptobacillus, Veillonella, Vibrio, and Yersinia species.
- 3. A method of identifying an antimicrobial compound comprising the steps of:(a) contacting microbial cells with a test compound and with a membrane potential indicator dye, and (b) detecting an increasing accumulation of said dye and retention of dye despite reduction of microbial cell viability wherein the microbial cells are gram-positive bacteria selected from the group consisting of Staphylococcus, Streptococcus, Micrococcus, Peptococcus, Peptostreptococcus, Enterococcus, Bacillus, Clostridium, Lactobacillus, Listeria, Erysipelothrix, Propionibacterium, Eubacterium, and Corynebacterium species.
- 4. The method of any one of claims 1 through 3 further comprising the step of determining microbial cell viability.
- 5. The method of any one of claims 1 through 3 further comprising the steps of:(a) contacting a mammalian cell with said test compound and with said membrane potential indicator dye, and (b) detecting no substantial change in dye uptake.
- 6. The method of any one of claims 1 through 3 further comprising the step of assaying said test compound for the ability to inhibit growth of microbial cells or to kill microbial cells.
- 7. The method of any one of claims 1 through 3 further comprising the steps of assaying said test compound for in vivo oral availability or oral activity.
- 8. The method of any one of claims 1 through 3 wherein said membrane potential indicator dye is DiOC6(3) (3,3′-dihexyloxacarbocyanine iodide).
- 9. The method of any one of claims 1 through 3 wherein said membrane potential indicator dye is JC-1 (5,5′,6,6′-tetrachloro-1,1′,3,3′-tetraethylbenzimidazolylcarbocyanine halide, or CBIC2(3)).
- 10. The method of any one of claims 1 through 3 wherein said membrane potential indicator dye is dihydrorhodamine 123.
- 11. The method of any one of claims 1 through 3 wherein said membrane potential indicator dye is MitoTracker® Red CM-H2Xros (M-7513; C32H33ClN2O).
Parent Case Info
This application is a continuation of U.S. Ser. No. 09/404,926, filed Sep. 24, 1999, which claims priority based on U.S. Provisional Application Serial No. 60/143,485 filed Jul. 12, 1999, U.S. Provisional Application Serial No. 60/109,905 filed Nov. 25, 1998, and U.S. Provisional Application Serial No. 60/101,778 filed Sep. 25, 1998.
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Number |
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Date |
Kind |
4859584 |
Horan et al. |
Aug 1989 |
A |
6143516 |
Little, II et al. |
Nov 2000 |
A |
Non-Patent Literature Citations (5)
Entry |
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Provisional Applications (3)
|
Number |
Date |
Country |
|
60/143485 |
Jul 1999 |
US |
|
60/109905 |
Nov 1998 |
US |
|
60/101778 |
Sep 1998 |
US |
Continuations (1)
|
Number |
Date |
Country |
Parent |
09/404926 |
Sep 1999 |
US |
Child |
09/626995 |
|
US |