Development of a Fluorescence-based Trypanosoma cruzi CYP51 Inhibition Assay for Effective Compound Triaging in Drug Discovery Programmes for Chagas Disease
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{"title"=>"Development of a Fluorescence-based Trypanosoma cruzi CYP51 Inhibition Assay for Effective Compound Triaging in Drug Discovery Programmes for Chagas Disease", "type"=>"journal", "authors"=>[{"first_name"=>"Jennifer", "last_name"=>"Riley", "scopus_author_id"=>"55808385500"}, {"first_name"=>"Stephen", "last_name"=>"Brand", "scopus_author_id"=>"56888422600"}, {"first_name"=>"Michael", "last_name"=>"Voice", "scopus_author_id"=>"6602100653"}, {"first_name"=>"Ivan", "last_name"=>"Caballero", "scopus_author_id"=>"55249415400"}, {"first_name"=>"David", "last_name"=>"Calvo", "scopus_author_id"=>"57201021692"}, {"first_name"=>"Kevin D.", "last_name"=>"Read", "scopus_author_id"=>"23567129400"}], "year"=>2015, "source"=>"PLoS Neglected Tropical Diseases", "identifiers"=>{"pmid"=>"26394211", "doi"=>"10.1371/journal.pntd.0004014", "sgr"=>"84943176065", "scopus"=>"2-s2.0-84943176065", "issn"=>"19352735", "pui"=>"606211319"}, "id"=>"f5849615-fb85-38ea-b33d-548d4ebe66f7", "abstract"=>"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.[on SciFinder (R)]", "link"=>"http://www.mendeley.com/research/development-fluorescencebased-trypanosoma-cruzi-cyp51-inhibition-assay-effective-compound-triaging-d", "reader_count"=>32, "reader_count_by_academic_status"=>{"Unspecified"=>3, "Professor > Associate Professor"=>1, "Student > Doctoral Student"=>2, "Researcher"=>9, "Student > Ph. D. Student"=>3, "Student > Master"=>6, "Other"=>2, "Student > Bachelor"=>4, "Professor"=>2}, "reader_count_by_user_role"=>{"Unspecified"=>3, "Professor > Associate Professor"=>1, "Student > Doctoral Student"=>2, "Researcher"=>9, "Student > Ph. D. Student"=>3, "Student > Master"=>6, "Other"=>2, "Student > Bachelor"=>4, "Professor"=>2}, "reader_count_by_subject_area"=>{"Unspecified"=>4, "Biochemistry, Genetics and Molecular Biology"=>5, "Agricultural and Biological Sciences"=>5, "Medicine and Dentistry"=>2, "Veterinary Science and Veterinary Medicine"=>1, "Pharmacology, Toxicology and Pharmaceutical Science"=>3, "Chemistry"=>10, "Immunology and Microbiology"=>1, "Decision Sciences"=>1}, "reader_count_by_subdiscipline"=>{"Medicine and Dentistry"=>{"Medicine and Dentistry"=>2}, "Chemistry"=>{"Chemistry"=>10}, "Decision Sciences"=>{"Decision Sciences"=>1}, "Immunology and Microbiology"=>{"Immunology and Microbiology"=>1}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>5}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>5}, "Unspecified"=>{"Unspecified"=>4}, "Pharmacology, Toxicology and Pharmaceutical Science"=>{"Pharmacology, Toxicology and Pharmaceutical Science"=>3}, "Veterinary Science and Veterinary Medicine"=>{"Veterinary Science and Veterinary Medicine"=>1}}, "reader_count_by_country"=>{"Argentina"=>1, "United States"=>1, "Uruguay"=>1}, "group_count"=>1}

Scopus | Further Information

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Figshare

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  • {"files"=>["https://ndownloader.figshare.com/files/2283297"], "description"=>"<p>a: Traditional endogenous substrate (Lanosterol) and CYP51 specific metabolite (FF-MAS); b: Fluorogenic probe substrate BOMCC is metabolised to CHC by CYP51.</p>", "links"=>[], "tags"=>["Chagas Disease Chagas disease", "assay", "parasite Trypanosoma cruzi", "Cyp 51", "phenotypic screening campaign", "cruzi CYP 51", "CYP 51 inhibition", "Drug Discovery Programmes", "Neglected Diseases initiative", "drug discovery portfolio"], "article_id"=>1552173, "categories"=>["Biological Sciences"], "users"=>["Jennifer Riley", "Stephen Brand", "Michael Voice", "Ivan Caballero", "David Calvo", "Kevin D. Read"], "doi"=>"https://dx.doi.org/10.1371/journal.pntd.0004014.g002", "stats"=>{"downloads"=>4, "page_views"=>28, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Development_of_a_Fluorescence_based_Trypanosoma_cruzi_Fig_2_CYP51_Inhibition_Assay_for_Effective_Compound_Triaging_in_Drug_Discovery_Programmes_for_Chagas_Disease_/1552173", "title"=>"Development of a Fluorescence-based <i>Trypanosoma cruzi - Fig 2 </i> CYP51 Inhibition Assay for Effective Compound Triaging in Drug Discovery Programmes for Chagas Disease", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2015-09-22 02:51:35"}
  • {"files"=>["https://ndownloader.figshare.com/files/2283301"], "description"=>"<p>a: Linearity of CHC production (expressed as average rate of fluorescence units (AFU)) with increasing T. cruzi CYP51 enzyme concentration (96 well format; average points (n = 3) with error bars); b: Michaelis-Menten Kinetic determination (96 well format; average points (n = 3) with error bars).</p>", "links"=>[], "tags"=>["Chagas Disease Chagas disease", "assay", "parasite Trypanosoma cruzi", "Cyp 51", "phenotypic screening campaign", "cruzi CYP 51", "CYP 51 inhibition", "Drug Discovery Programmes", "Neglected Diseases initiative", "drug discovery portfolio"], "article_id"=>1552177, "categories"=>["Biological Sciences"], "users"=>["Jennifer Riley", "Stephen Brand", "Michael Voice", "Ivan Caballero", "David Calvo", "Kevin D. Read"], "doi"=>"https://dx.doi.org/10.1371/journal.pntd.0004014.g003", "stats"=>{"downloads"=>2, "page_views"=>15, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Development_of_a_Fluorescence_based_Trypanosoma_cruzi_Fig_3_CYP51_Inhibition_Assay_for_Effective_Compound_Triaging_in_Drug_Discovery_Programmes_for_Chagas_Disease_/1552177", "title"=>"Development of a Fluorescence-based <i>Trypanosoma cruzi - Fig 3 </i> CYP51 Inhibition Assay for Effective Compound Triaging in Drug Discovery Programmes for Chagas Disease", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2015-09-22 02:51:35"}
  • {"files"=>["https://ndownloader.figshare.com/files/2283303"], "description"=>"<p>a: The effect of solvent on <i>T</i>. <i>cruzi</i> CYP51 activity; b: Cytochrome C reductase activity of recombinant preparations.</p>", "links"=>[], "tags"=>["Chagas Disease Chagas disease", "assay", "parasite Trypanosoma cruzi", "Cyp 51", "phenotypic screening campaign", "cruzi CYP 51", "CYP 51 inhibition", "Drug Discovery Programmes", "Neglected Diseases initiative", "drug discovery portfolio"], "article_id"=>1552179, "categories"=>["Biological Sciences"], "users"=>["Jennifer Riley", "Stephen Brand", "Michael Voice", "Ivan Caballero", "David Calvo", "Kevin D. Read"], "doi"=>"https://dx.doi.org/10.1371/journal.pntd.0004014.g004", "stats"=>{"downloads"=>3, "page_views"=>28, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Development_of_a_Fluorescence_based_Trypanosoma_cruzi_Fig_4_CYP51_Inhibition_Assay_for_Effective_Compound_Triaging_in_Drug_Discovery_Programmes_for_Chagas_Disease_/1552179", "title"=>"Development of a Fluorescence-based <i>Trypanosoma cruzi - Fig 4 </i> CYP51 Inhibition Assay for Effective Compound Triaging in Drug Discovery Programmes for Chagas Disease", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2015-09-22 02:51:35"}
  • {"files"=>["https://ndownloader.figshare.com/files/2283307"], "description"=>"<p>a: <i>T</i>. <i>cruzi</i> CYP51 inhibition plotted against <i>T</i>. <i>cruzi</i> activity for compounds from the different chemical series (coloured green), in addition to 16 known inhibitors of human CYP51 used as a reference set to define the correlation curve (coloured red). <i>T</i>. <i>cruzi</i> active series within the dotted box are prioritised for further medicinal chemistry; b: <i>T</i>. <i>cruzi</i> CYP51 inhibition plotted against <i>T</i>. <i>cruzi</i> activity for series one (green), and series two (blue) compared with the human CYP51 training set (red); c: <i>T</i>. <i>cruzi</i> CYP51 inhibition plotted against <i>T</i>. <i>cruzi</i> activity for a third series (yellow) compared with the human CYP51 training set (red). pEC<sub>50</sub> or pIC<sub>50</sub> is the −log of EC<sub>50</sub> or IC<sub>50</sub> respectively in M.</p>", "links"=>[], "tags"=>["Chagas Disease Chagas disease", "assay", "parasite Trypanosoma cruzi", "Cyp 51", "phenotypic screening campaign", "cruzi CYP 51", "CYP 51 inhibition", "Drug Discovery Programmes", "Neglected Diseases initiative", "drug discovery portfolio"], "article_id"=>1552181, "categories"=>["Biological Sciences"], "users"=>["Jennifer Riley", "Stephen Brand", "Michael Voice", "Ivan Caballero", "David Calvo", "Kevin D. Read"], "doi"=>"https://dx.doi.org/10.1371/journal.pntd.0004014.g005", "stats"=>{"downloads"=>2, "page_views"=>12, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Development_of_a_Fluorescence_based_Trypanosoma_cruzi_Fig_5_CYP51_Inhibition_Assay_for_Effective_Compound_Triaging_in_Drug_Discovery_Programmes_for_Chagas_Disease_/1552181", "title"=>"Development of a Fluorescence-based <i>Trypanosoma cruzi - Fig 5 </i> CYP51 Inhibition Assay for Effective Compound Triaging in Drug Discovery Programmes for Chagas Disease", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2015-09-22 02:51:35"}
  • {"files"=>["https://ndownloader.figshare.com/files/2283310"], "description"=>"<p>Chemical moieties associated with CYP inhibition.</p>", "links"=>[], "tags"=>["Chagas Disease Chagas disease", "assay", "parasite Trypanosoma cruzi", "Cyp 51", "phenotypic screening campaign", "cruzi CYP 51", "CYP 51 inhibition", "Drug Discovery Programmes", "Neglected Diseases initiative", "drug discovery portfolio"], "article_id"=>1552182, "categories"=>["Biological Sciences"], "users"=>["Jennifer Riley", "Stephen Brand", "Michael Voice", "Ivan Caballero", "David Calvo", "Kevin D. Read"], "doi"=>"https://dx.doi.org/10.1371/journal.pntd.0004014.g006", "stats"=>{"downloads"=>2, "page_views"=>19, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Chemical_moieties_associated_with_CYP_inhibition_/1552182", "title"=>"Chemical moieties associated with CYP inhibition.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2015-09-22 02:51:35"}
  • {"files"=>["https://ndownloader.figshare.com/files/2283311"], "description"=>"<p>Error bars are the standard deviations of 16 replicates for each control and plate.</p>", "links"=>[], "tags"=>["Chagas Disease Chagas disease", "assay", "parasite Trypanosoma cruzi", "Cyp 51", "phenotypic screening campaign", "cruzi CYP 51", "CYP 51 inhibition", "Drug Discovery Programmes", "Neglected Diseases initiative", "drug discovery portfolio"], "article_id"=>1552183, "categories"=>["Biological Sciences"], "users"=>["Jennifer Riley", "Stephen Brand", "Michael Voice", "Ivan Caballero", "David Calvo", "Kevin D. Read"], "doi"=>"https://dx.doi.org/10.1371/journal.pntd.0004014.g007", "stats"=>{"downloads"=>2, "page_views"=>16, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Trends_of_control_1_closed_circles_solvent_only_control_0_inhibition_control_2_open_circles_reaction_in_absence_of_bactosomes_100_inhibition_and_Z_grey_squares_of_representative_high_throughput_screening_runs_of_T_cruzi_CYP51_FLINT_a_and_cytochrome_c_red/1552183", "title"=>"Trends of control 1 (closed circles, solvent only control, 0% inhibition), control 2 (open circles, reaction in absence of bactosomes, 100% inhibition) and Z’ (grey squares) of representative high-throughput screening runs of <i>T</i>. <i>cruzi</i> CYP51 FLINT (a) and cytochrome c reductase absorbance (b) assays in 384 well format.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2015-09-22 02:51:35"}
  • {"files"=>["https://ndownloader.figshare.com/files/2283312"], "description"=>"<p><sup><b>a</b></sup> Coefficient of variation for all IC<sub>50</sub>’s were lower than 1%</p><p>IC<sub>50</sub> Determination of a test set of compounds.</p>", "links"=>[], "tags"=>["Chagas Disease Chagas disease", "assay", "parasite Trypanosoma cruzi", "Cyp 51", "phenotypic screening campaign", "cruzi CYP 51", "CYP 51 inhibition", "Drug Discovery Programmes", "Neglected Diseases initiative", "drug discovery portfolio"], "article_id"=>1552184, "categories"=>["Biological Sciences"], "users"=>["Jennifer Riley", "Stephen Brand", "Michael Voice", "Ivan Caballero", "David Calvo", "Kevin D. Read"], "doi"=>"https://dx.doi.org/10.1371/journal.pntd.0004014.t001", "stats"=>{"downloads"=>4, "page_views"=>16, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_IC_50_Determination_of_a_test_set_of_compounds_/1552184", "title"=>"IC<sub>50</sub> Determination of a test set of compounds.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2015-09-22 02:51:35"}

PMC Usage Stats | Further Information

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Relative Metric

{"start_date"=>"2015-01-01T00:00:00Z", "end_date"=>"2015-12-31T00:00:00Z", "subject_areas"=>[]}
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