Development of a Fluorescence-based Trypanosoma cruzi CYP51 Inhibition Assay for Effective Compound Triaging in Drug Discovery Programmes for Chagas Disease.
Chagas disease, caused by the protozoan parasite Trypanosoma cruzi (T. cruzi), is a life threatening global health problem with only two drugs available for treatment (benznidazole and nifurtimox), both having variable efficacy in the chronic stage of the disease and high rates of adverse drug react...
Saved in:
| Main Authors: | , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2015-09-01
|
| Series: | PLoS Neglected Tropical Diseases |
| Online Access: | https://journals.plos.org/plosntds/article/file?id=10.1371/journal.pntd.0004014&type=printable |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1849331852696879104 |
|---|---|
| author | Jennifer Riley Stephen Brand Michael Voice Ivan Caballero David Calvo Kevin D Read |
| author_facet | Jennifer Riley Stephen Brand Michael Voice Ivan Caballero David Calvo Kevin D Read |
| author_sort | Jennifer Riley |
| collection | DOAJ |
| description | Chagas disease, caused by the protozoan parasite Trypanosoma cruzi (T. cruzi), is a life threatening global health problem with only two drugs available for treatment (benznidazole and nifurtimox), both having variable efficacy in the chronic stage of the disease and high rates of adverse drug reactions. Inhibitors of sterol 14α-demethylase (CYP51) have proven effective against T. cruzi in vitro and in vivo in animal models of Chagas disease. Consequently two azole inhibitors of CYP51 (posaconazole and ravuconazole) have recently entered clinical development by the Drugs for Neglected Diseases initiative. Further new drug treatments for this disease are however still urgently required, particularly having a different mode of action to CYP51 in order to balance the overall risk in the drug discovery portfolio. This need has now been further strengthened by the very recent reports of treatment failure in the clinic for both posaconazole and ravuconazole. To this end and to prevent enrichment of drug candidates against a single target, there is a clear need for a robust high throughput assay for CYP51 inhibition in order to evaluate compounds active against T. cruzi arising from phenotypic screens. A high throughput fluorescence based functional assay using recombinantly expressed T. cruzi CYP51 (Tulahuen strain) is presented here that meets this requirement. This assay has proved valuable in prioritising medicinal chemistry resource on only those T. cruzi active series arising from a phenotypic screening campaign where it is clear that the predominant mode of action is likely not via inhibition of CYP51. |
| format | Article |
| id | doaj-art-e1fba0f7f43045deb3d5a8e5c05870d0 |
| institution | Kabale University |
| issn | 1935-2727 1935-2735 |
| language | English |
| publishDate | 2015-09-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS Neglected Tropical Diseases |
| spelling | doaj-art-e1fba0f7f43045deb3d5a8e5c05870d02025-08-20T03:46:23ZengPublic Library of Science (PLoS)PLoS Neglected Tropical Diseases1935-27271935-27352015-09-0199e000401410.1371/journal.pntd.0004014Development of a Fluorescence-based Trypanosoma cruzi CYP51 Inhibition Assay for Effective Compound Triaging in Drug Discovery Programmes for Chagas Disease.Jennifer RileyStephen BrandMichael VoiceIvan CaballeroDavid CalvoKevin D ReadChagas disease, caused by the protozoan parasite Trypanosoma cruzi (T. cruzi), is a life threatening global health problem with only two drugs available for treatment (benznidazole and nifurtimox), both having variable efficacy in the chronic stage of the disease and high rates of adverse drug reactions. Inhibitors of sterol 14α-demethylase (CYP51) have proven effective against T. cruzi in vitro and in vivo in animal models of Chagas disease. Consequently two azole inhibitors of CYP51 (posaconazole and ravuconazole) have recently entered clinical development by the Drugs for Neglected Diseases initiative. Further new drug treatments for this disease are however still urgently required, particularly having a different mode of action to CYP51 in order to balance the overall risk in the drug discovery portfolio. This need has now been further strengthened by the very recent reports of treatment failure in the clinic for both posaconazole and ravuconazole. To this end and to prevent enrichment of drug candidates against a single target, there is a clear need for a robust high throughput assay for CYP51 inhibition in order to evaluate compounds active against T. cruzi arising from phenotypic screens. A high throughput fluorescence based functional assay using recombinantly expressed T. cruzi CYP51 (Tulahuen strain) is presented here that meets this requirement. This assay has proved valuable in prioritising medicinal chemistry resource on only those T. cruzi active series arising from a phenotypic screening campaign where it is clear that the predominant mode of action is likely not via inhibition of CYP51.https://journals.plos.org/plosntds/article/file?id=10.1371/journal.pntd.0004014&type=printable |
| spellingShingle | Jennifer Riley Stephen Brand Michael Voice Ivan Caballero David Calvo Kevin D Read Development of a Fluorescence-based Trypanosoma cruzi CYP51 Inhibition Assay for Effective Compound Triaging in Drug Discovery Programmes for Chagas Disease. PLoS Neglected Tropical Diseases |
| title | Development of a Fluorescence-based Trypanosoma cruzi CYP51 Inhibition Assay for Effective Compound Triaging in Drug Discovery Programmes for Chagas Disease. |
| title_full | Development of a Fluorescence-based Trypanosoma cruzi CYP51 Inhibition Assay for Effective Compound Triaging in Drug Discovery Programmes for Chagas Disease. |
| title_fullStr | Development of a Fluorescence-based Trypanosoma cruzi CYP51 Inhibition Assay for Effective Compound Triaging in Drug Discovery Programmes for Chagas Disease. |
| title_full_unstemmed | Development of a Fluorescence-based Trypanosoma cruzi CYP51 Inhibition Assay for Effective Compound Triaging in Drug Discovery Programmes for Chagas Disease. |
| title_short | Development of a Fluorescence-based Trypanosoma cruzi CYP51 Inhibition Assay for Effective Compound Triaging in Drug Discovery Programmes for Chagas Disease. |
| title_sort | development of a fluorescence based trypanosoma cruzi cyp51 inhibition assay for effective compound triaging in drug discovery programmes for chagas disease |
| url | https://journals.plos.org/plosntds/article/file?id=10.1371/journal.pntd.0004014&type=printable |
| work_keys_str_mv | AT jenniferriley developmentofafluorescencebasedtrypanosomacruzicyp51inhibitionassayforeffectivecompoundtriagingindrugdiscoveryprogrammesforchagasdisease AT stephenbrand developmentofafluorescencebasedtrypanosomacruzicyp51inhibitionassayforeffectivecompoundtriagingindrugdiscoveryprogrammesforchagasdisease AT michaelvoice developmentofafluorescencebasedtrypanosomacruzicyp51inhibitionassayforeffectivecompoundtriagingindrugdiscoveryprogrammesforchagasdisease AT ivancaballero developmentofafluorescencebasedtrypanosomacruzicyp51inhibitionassayforeffectivecompoundtriagingindrugdiscoveryprogrammesforchagasdisease AT davidcalvo developmentofafluorescencebasedtrypanosomacruzicyp51inhibitionassayforeffectivecompoundtriagingindrugdiscoveryprogrammesforchagasdisease AT kevindread developmentofafluorescencebasedtrypanosomacruzicyp51inhibitionassayforeffectivecompoundtriagingindrugdiscoveryprogrammesforchagasdisease |