A New Approach to Drug Discovery: High-Throughput Screening of Microbial Natural Extracts against Aspergillus fumigatus Using Resazurin

Monteiro, Maria Cândida; Cruz, Mercedes de la; Cantizani, Juan; Moreno, Catalina; Tormo, José R.; Mellado, Emilia; Lucas, J. Ramón De; Asensio, Francisco; Valiante, Vito; Brakhage, Axel A.; Latgé, Jean‐Paul; Genilloud, Olga; Vicente, Francisca

doi:10.1177/1087057111433459

Cited by 124 publications

(86 citation statements)

References 17 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Culture plates were incubated at 37°C for 30 h. The variation of the color of resazurin was visually observed after the incubation of the plates, and the growth inhibition was quantified by measuring the fluorescence as described above (12). All of the experiments were performed in triplicate.…”

Section: Methodsmentioning

confidence: 99%

“…Cell viability of A. fumigatus was measured using resazurin as described previously (12). In the case of C. albicans, yeast suspensions from cryovials were streaked on SDA plates and incubated for 18 h at 37°C, and colonies were inoculated in 10 ml Sabouraud dextrose broth (SDB).…”

Section: Methodsmentioning

confidence: 99%

“…An HTS approach was used to test, in combination with CAS, 20,000 microbial natural product extracts from actinomycetes and fungi (available from the Fundación MEDINA collection) (12). The result was that 0.94% of extracts analyzed displayed a CAS-enhancing activity against the two A. fumigatus wild-type strains tested, ATCC 46645 and CEA17⌬akuB (inhibition cutoff of Ն60%) (23,24).…”

Section: Isolation and Structure Elucidation Of Humidimycin (Mdn-0010mentioning

confidence: 99%

See 2 more Smart Citations

Hitting the Caspofungin Salvage Pathway of Human-Pathogenic Fungi with the Novel Lasso Peptide Humidimycin (MDN-0010)

Valiante

Monteiro

Martín

et al. 2015

Antimicrob Agents Chemother

Self Cite

View full text Add to dashboard Cite

f Fungal infections have increased dramatically in the last 2 decades, and fighting infectious diseases requires innovative approaches such as the combination of two drugs acting on different targets or even targeting a salvage pathway of one of the drugs. The fungal cell wall biosynthesis is inhibited by the clinically used antifungal drug caspofungin. This antifungal activity has been found to be potentiated by humidimycin, a new natural product identified from the screening of a collection of 20,000 microbial extracts, which has no major effect when used alone. An analysis of transcriptomes and selected Aspergillus fumigatus mutants indicated that humidimycin affects the high osmolarity glycerol response pathway. By combining humidimycin and caspofungin, a strong increase in caspofungin efficacy was achieved, demonstrating that targeting different signaling pathways provides an excellent basis to develop novel anti-infective strategies.

show abstract

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Isolation and Structure Elucidation Of Humidimycin (Mdn-0010mentioning

confidence: 99%

See 1 more Smart Citation

Hitting the Caspofungin Salvage Pathway of Human-Pathogenic Fungi with the Novel Lasso Peptide Humidimycin (MDN-0010)

Valiante

Monteiro

Martín

et al. 2015

Antimicrob Agents Chemother

Self Cite

View full text Add to dashboard Cite

show abstract

“…After microbial incubation, growth is measured. The methods usually used for this quantification are colorimetric, such as tetrazolium salts, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) or resazurin dye [45]. …”

Section: Methodologies Of Screeningmentioning

confidence: 99%

Current Screening Methodologies in Drug Discovery for Selected Human Diseases

et al. 2018

Self Cite

View full text Add to dashboard Cite

The increase of many deadly diseases like infections by multidrug-resistant bacteria implies re-inventing the wheel on drug discovery. A better comprehension of the metabolisms and regulation of diseases, the increase in knowledge based on the study of disease-born microorganisms’ genomes, the development of more representative disease models and improvement of techniques, technologies, and computation applied to biology are advances that will foster drug discovery in upcoming years. In this paper, several aspects of current methodologies for drug discovery of antibacterial and antifungals, anti-tropical diseases, antibiofilm and antiquorum sensing, anticancer and neuroprotectors are considered. For drug discovery, two different complementary approaches can be applied: classical pharmacology, also known as phenotypic drug discovery, which is the historical basis of drug discovery, and reverse pharmacology, also designated target-based drug discovery. Screening methods based on phenotypic drug discovery have been used to discover new natural products mainly from terrestrial origin. Examples of the discovery of marine natural products are provided. A section on future trends provides a comprehensive overview on recent advances that will foster the pharmaceutical industry.

show abstract

“…After screening of 39,000 crude extracts, 49 were found to block fermentation, and three were characterized as novel broad-spectrum antibiotics. An alternative colorimetric assay utilizes resazurin, a general indicator of metabolic activity, to detect antifungal activity against the human pathogen A. fumigatus [16]. High throughput screens can also be carried out against engineered assay strains, as exemplified in a screen for activators of the heat shock response in human cells [17].…”

Section: Top-down Approachesmentioning

confidence: 99%

Recent advances in natural product discovery

Luo

Cobb

Zhao

2014

Current Opinion in Biotechnology

133

112

View full text Add to dashboard Cite

Natural products have been and continue to be the source and inspiration for a substantial fraction of human therapeutics. Although the pharmaceutical industry has largely turned its back on natural product discovery efforts, such efforts continue to flourish in academia with promising results. Natural products have traditionally been identified from a top-down perspective, but more recently genomics- and bioinformatics-guided bottom-up approaches have provided powerful alternative strategies. Here we review recent advances in natural product discovery from both angles, including diverse sampling and innovative culturing and screening approaches, as well as genomics-driven discovery and genetic manipulation techniques for both native and heterologous expression.

show abstract

A New Approach to Drug Discovery: High-Throughput Screening of Microbial Natural Extracts against Aspergillus fumigatus Using Resazurin

Cited by 124 publications

References 17 publications

Hitting the Caspofungin Salvage Pathway of Human-Pathogenic Fungi with the Novel Lasso Peptide Humidimycin (MDN-0010)

Hitting the Caspofungin Salvage Pathway of Human-Pathogenic Fungi with the Novel Lasso Peptide Humidimycin (MDN-0010)

Current Screening Methodologies in Drug Discovery for Selected Human Diseases

Recent advances in natural product discovery

Contact Info

Product

Resources

About