c5cdb4a1a71843c01821a80f63f60d60 pmc_11514582.pdf d4bd59470afbe5d03d12476b4d3162a058ccc150 pmc_11514582.pdf eecf8908c484f9f5422a0f2a425fe7df297417dcbd19eff36411e6911affbee0 pmc_11514582.pdf Title: Leveraging the Aggregated Protein Dye YAT2150 for Malaria Chemotherapy Subject: Background/Objectives: YAT2150 is a first-in-class antiplasmodial compound that has been recently proposed as a new interesting drug for malaria therapy. Methods/Results: The fluorescence of YAT2150 rapidly increases upon its entry into Plasmodium, a property that can be of use for the design of highly sensitive diagnostic approaches. YAT2150 blocks the in vitro development of the ookinete stage of Plasmodium and, when added to an infected blood meal, inhibits oocyst formation in the mosquito. Thus, the compound could possibly contribute to future transmission-blocking antimalarial strategies. Cell influx/efflux studies in Caco-2 cells suggest that YAT2150 is internalized by endocytosis and also through the OATP2B1 transporter, whereas its main export route would be via OST. YAT2150 has an overall favorable drug metabolism and pharmacokinetics profile, and its moderate cytotoxicity can be significantly reduced upon encapsulation in immunoliposomes, which leads to a dramatic increase in the drug selectivity index to values close to 1000. Although YAT2150 binds amyloid-forming peptides, its in vitro fluorescence emission is stronger upon association with peptides that form amorphous aggregates, suggesting that regions enriched in unstructured proteins are the preferential binding sites of the drug inside Plasmodium cells. The reduction of protein aggregation in the parasite after YAT2150 treatment, which has been suggested to be directly related to the drug’s mode of action, is also observed following treatment with quinoline antimalarials like chloroquine and primaquine. Conclusions: Altogether, the data presented here indicate that YAT2150 can represent the spearhead of a new family of compounds for malaria diagnosis and therapy due to its presumed novel mode of action based on the interaction with functional protein aggregates in the pathogen. Keywords: Plasmodium falciparum; malaria; protein aggregation; YAT2150 Author: Claudia Camarero-Hoyos, Inés Bouzón-Arnáiz, Yunuen Avalos-Padilla, Antonino Nicolò Fallica, Lucía Román-Álamo, Miriam Ramírez, Emma Portabella, Ona Cuspinera, Daniela Currea-Ayala, Marc Orozco-Quer, Maria Ribera, Inga Siden-Kiamos, Lefteris Spanos, Valentín Iglesias, Benigno Crespo, Sara Viera, David Andreu, Elena Sulleiro, Francesc Zarzuela, Nerea Urtasun, Sandra Pérez-Torras, Marçal Pastor-Anglada, Elsa M. Arce, Diego Muñoz-Torrero and Xavier Fernàndez-Busquets Creator: LaTeX with hyperref Producer: pdfTeX-1.40.25 CreationDate: Tue Oct 8 12:43:13 2024 CEST ModDate: Tue Oct 8 13:06:28 2024 CEST Custom Metadata: no Metadata Stream: no Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 23 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 6437629 bytes Optimized: no PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- WHZOOC+VnURWPalladioL-Bold Type 1 Custom yes yes yes 10 0 RFQTET+URWPalladioL-Bold Type 1 Custom yes yes yes 16 0 PHQTUW+URWPalladioL-Roma Type 1 Custom yes yes yes 22 0 QIMCOO+URWPalladioL-Ital Type 1 Custom yes yes yes 27 0 BBAXXJ+VnURWPalladioL Type 1 Custom yes yes yes 32 0 IAFAOG+TimesNewRomanPS-BoldItalicMT Type 1C WinAnsi yes yes no 49 0 NXQZWG+EURM10 Type 1 Builtin yes yes yes 72 0 PVZWPK+CMSY10 Type 1 Builtin yes yes yes 80 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NLPMBL+PalatinoLinotype TrueType MacRoman yes yes no 268 0 NLPINK+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 302 0 NLPINL+PalatinoLinotype CID TrueType Identity-H yes yes yes 305 0 NLPINM+PalatinoLinotype TrueType WinAnsi yes yes no 311 0 NLPINO+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 314 0 NLPMBL+PalatinoLinotype TrueType MacRoman yes yes no 317 0 Jhove (Rel. 1.28.0, 2023-05-18) Date: 2025-02-06 02:46:48 CET RepresentationInformation: pmc_11514582.pdf ReportingModule: PDF-hul, Rel. 1.12.4 (2023-03-16) LastModified: 2025-02-05 02:55:06 CET Size: 6437629 Format: PDF Version: 1.7 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 612 FreeObjects: 1 IncrementalUpdates: 0 DocumentCatalog: PageLayout: SinglePage PageMode: UseNone Outlines: Item: Title: Introduction Destination: section.1 Item: Title: Materials and Methods Destination: section.2 Children: Item: Title: Reagents Destination: subsection.2.1 Item: Title: Prediction of Prion-like Propensities Destination: subsection.2.2 Item: Title: Microscopic Observation of Clinical Samples Destination: subsection.2.3 Item: Title: Assay of Gametocyte-to-Ookinete Transition Inhibition Destination: subsection.2.4 Item: Title: Membrane Blood Feeding Assay Destination: subsection.2.5 Item: Title: Cytochrome Inhibition Activity Assay Destination: subsection.2.6 Item: Title: Cytotoxicity Assays Destination: subsection.2.7 Item: Title: Human Plasma Protein Binding Determination Destination: subsection.2.8 Item: Title: Influx and Efflux Assays in Caco-2 Cells Destination: subsection.2.9 Item: Title: Treatment of P. falciparum Cultures Destination: subsection.2.10 Item: Title: In Vitro Peptide Aggregation Assays Destination: subsection.2.11 Item: Title: Transmission Electron Microscopy Destination: subsection.2.12 Item: Title: Protein Aggregation Assays in P. falciparum Cultures Destination: subsection.2.13 Item: Title: In Vivo Toxicity Determination Destination: subsection.2.14 Item: Title: Ethical Issues Destination: subsection.2.15 Item: Title: Statistical Data Analysis Destination: subsection.2.16 Item: Title: Results Destination: section.3 Children: Item: Title: Rapid Detection of Malaria Infection in YAT2150-Stained Clinical Samples Destination: subsection.3.1 Item: Title: Transmission Blocking Assays Destination: subsection.3.2 Item: Title: Drug Metabolism and Pharmacokinetics (DMPK) and Early Safety Profiling of YAT2150 Destination: subsection.3.3 Item: Title: Cell Influx and Efflux Studies Destination: subsection.3.4 Item: Title: In Vitro Antiplasmodial Activity of YAT2150 Encapsulated in Liposomes Destination: subsection.3.5 Item: Title: Effect of Clinically Used Antimalarial Drugs on the Level of Protein Aggregation in P. falciparum Cultures Destination: subsection.3.6 Item: Title: Interaction of YAT2150 with Aggregative Peptides Destination: subsection.3.7 Item: Title: Discussion Destination: section.4 Item: Title: Conclusions Destination: section.5 Item: Title: References Destination: appendix.A. Info: Title: Leveraging the Aggregated Protein Dye YAT2150 for Malaria Chemotherapy Author: Claudia Camarero-Hoyos, Inés Bouzón-Arnáiz, Yunuen Avalos-Padilla, Antonino Nicolò Fallica, Lucía Román-Álamo, Miriam Ramírez, Emma Portabella, Ona Cuspinera, Daniela Currea-Ayala, Marc Orozco-Quer, Maria Ribera, Inga Siden-Kiamos, Lefteris Spanos, Valentín Iglesias, Benigno Crespo, Sara Viera, David Andreu, Elena Sulleiro, Francesc Zarzuela, Nerea Urtasun, Sandra Pérez-Torras, Marçal Pastor-Anglada, Elsa M. Arce, Diego Muñoz-Torrero and Xavier Fernàndez-Busquets Subject: Background/Objectives: YAT2150 is a first-in-class antiplasmodial compound that has been recently proposed as a new interesting drug for malaria therapy. Methods/Results: The fluorescence of YAT2150 rapidly increases upon its entry into Plasmodium, a property that can be of use for the design of highly sensitive diagnostic approaches. YAT2150 blocks the in vitro development of the ookinete stage of Plasmodium and, when added to an infected blood meal, inhibits oocyst formation in the mosquito. Thus, the compound could possibly contribute to future transmission-blocking antimalarial strategies. Cell influx/efflux studies in Caco-2 cells suggest that YAT2150 is internalized by endocytosis and also through the OATP2B1 transporter, whereas its main export route would be via OST. YAT2150 has an overall favorable drug metabolism and pharmacokinetics profile, and its moderate cytotoxicity can be significantly reduced upon encapsulation in immunoliposomes, which leads to a dramatic increase in the drug selectivity index to values close to 1000. Although YAT2150 binds amyloid-forming peptides, its in vitro fluorescence emission is stronger upon association with peptides that form amorphous aggregates, suggesting that regions enriched in unstructured proteins are the preferential binding sites of the drug inside Plasmodium cells. The reduction of protein aggregation in the parasite after YAT2150 treatment, which has been suggested to be directly related to the drug’s mode of action, is also observed following treatment with quinoline antimalarials like chloroquine and primaquine. Conclusions: Altogether, the data presented here indicate that YAT2150 can represent the spearhead of a new family of compounds for malaria diagnosis and therapy due to its presumed novel mode of action based on the interaction with functional protein aggregates in the pathogen. 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248, 719, 285, 729 Annotation: Subtype: Link Rect: 59, 684, 96, 694 Annotation: Subtype: Link Rect: 102, 684, 138, 694 Annotation: Subtype: Link Rect: 222, 649, 260, 660 Annotation: Subtype: Link Rect: 431, 614, 468, 625 Annotation: Subtype: Link Rect: 59, 581, 96, 591 Annotation: Subtype: Link Rect: 59, 546, 96, 556 Annotation: Subtype: Link Rect: 102, 546, 138, 556 Annotation: Subtype: Link Rect: 252, 523, 289, 533 Annotation: Subtype: Link Rect: 296, 523, 332, 533 Annotation: Subtype: Link Rect: 392, 499, 429, 510 Annotation: Subtype: Link Rect: 436, 499, 472, 510 Checksum: e75a22d6 Type: CRC32 Checksum: c5cdb4a1a71843c01821a80f63f60d60 Type: MD5 Checksum: d4bd59470afbe5d03d12476b4d3162a058ccc150 Type: SHA-1 Checksum: eecf8908c484f9f5422a0f2a425fe7df297417dcbd19eff36411e6911affbee0 Type: SHA-256