Prediction of Antileishmanial Compounds: General Model, Preparation, and Evaluation of 2-Acylpyrrole Derivatives DOI Creative Commons
C. Santiago, Bernabé Ortega-Tenezaca, Iratxe Barbolla

et al.

Journal of Chemical Information and Modeling, Journal Year: 2022, Volume and Issue: 62(16), P. 3928 - 3940

Published: Aug. 10, 2022

In this work, the SOFT.PTML tool has been used to pre-process a ChEMBL dataset of pre-clinical assays antileishmanial compound candidates. A comparative study different ML algorithms, such as logistic regression (LOGR), support vector machine (SVM), and random forests (RF), shown that IFPTML-LOGR model presents excellent values specificity sensitivity (81-98%) in training validation series. The use software illustrated with practical case focused on series 28 derivatives 2-acylpyrroles 5a,b, obtained through Pd(II)-catalyzed C-H radical acylation pyrroles. Their vitro leishmanicidal activity against visceral (L. donovani) cutaneous amazonensis) leishmaniasis was evaluated finding compounds 5bc (IC50 = 30.87 μM, SI > 10.17) 5bd 16.87 10.67) were approximately 6-fold more selective than drug reference (miltefosine) L. amazonensis promastigotes. addition, most showed low cytotoxicity, CC50 100 μg/mL J774 cells. Interestingly, IFPMTL-LOGR predicts correctly relative biological these acylpyrroles. computational high-throughput screening (cHTS) 5a,b performed calculating >20,700 scores vs large space 647 involving multiple Leishmania species, cell lines, potential target proteins. Overall, demonstrates all-in-one strategy is useful obtain IFPTML models friendly interface making work easier faster before. present also points new lead worthy further optimization hits.

Language: Английский

Anti-trypanosomatid drug discovery: progress and challenges DOI Creative Commons
Manu De Rycker, Susan Wyllie, David Horn

et al.

Nature Reviews Microbiology, Journal Year: 2022, Volume and Issue: 21(1), P. 35 - 50

Published: Aug. 22, 2022

Leishmaniasis (visceral and cutaneous), Chagas disease human African trypanosomiasis cause substantial death morbidity, particularly in low- middle-income countries. Although the situation has improved for trypanosomiasis, there remains an urgent need new medicines to treat leishmaniasis disease; clinical development pipeline is sparse disease. In this Review, we describe recent advances our understanding of biology causative pathogens, from drug discovery perspective, explore progress that been made candidates identification promising molecular targets. We also challenges developing discuss potential solutions overcome such hurdles. Gilbert colleagues kinetoplastid diseases how parasite affects process

Language: Английский

Citations

131
Challenges in drug discovery targeting TriTryp diseases with an emphasis on leishmaniasis DOI Creative Commons
Laura Maria Alcântara, Thalita Camêlo da Silva Ferreira, Fernanda Ramos Gadelha

et al.

International Journal for Parasitology Drugs and Drug Resistance, Journal Year: 2018, Volume and Issue: 8(3), P. 430 - 439

Published: Sept. 28, 2018

Tritryps diseases are devastating parasitic neglected infections caused by Leishmania spp., Trypanosoma cruzi and brucei subspecies. Together, these parasites affect more than 30 million people worldwide cause high mortality morbidity. Leishmaniasis comprises a complex group of with clinical manifestation ranging from cutaneous lesions to systemic visceral damage. Antimonials, the first-choice drugs used treat leishmaniasis, lead toxicity carry significant contraindications limiting its use. Drug-resistant parasite strains also matter for increasing concern, especially in areas very limited resources. The current scenario calls novel and/or improvement existing therapeutics as key research priorities field. Although several studies have shown advances drug discovery towards leishmaniasis recent years, knowledge gaps pipelines still need be addressed. In this review we discuss not only scientific non-scientific bottlenecks development, but central role public-private partnerships successful campaign treatment options against disease.

Language: Английский

Citations

114
Systematic functional analysis of Leishmania protein kinases identifies regulators of differentiation or survival DOI Creative Commons
Nicola Baker, Carolina Moura Costa Catta‐Preta, Rachel Neish

et al.

Nature Communications, Journal Year: 2021, Volume and Issue: 12(1)

Published: Feb. 23, 2021

Abstract Differentiation between distinct stages is fundamental for the life cycle of intracellular protozoan parasites and transmission hosts, requiring stringent spatial temporal regulation. Here, we apply kinome-wide gene deletion tagging in Leishmania mexicana promastigotes to define protein kinases with transition roles. Whilst 162 are dispensable, 44 kinase genes refractory likely core required parasite replication. Phenotyping pooled mutants using bar-seq projection pursuit clustering reveal functional phenotypic groups involved differentiation from metacyclic promastigote amastigote, growth survival macrophages mice, colonisation sand fly motility. This unbiased interrogation function allows targeted investigation organelle-associated signalling pathways successful parasitism.

Language: Английский

Citations

96
The current drug discovery landscape for trypanosomiasis and leishmaniasis: Challenges and strategies to identify drug targets DOI
Fernando Altamura, Rishi Rajesh, Carolina Moura Costa Catta‐Preta

et al.

Drug Development Research, Journal Year: 2020, Volume and Issue: 83(2), P. 225 - 252

Published: April 6, 2020

Abstract Human trypanosomiasis and leishmaniasis are vector‐borne neglected tropical diseases caused by infection with the protozoan parasites Trypanosoma spp. Leishmania spp., respectively. Once restricted to endemic areas, these now distributed worldwide due human migration, climate change, anthropogenic disturbance, causing significant health economic burden globally. The current chemotherapy used treat has limited efficacy, drug resistance is spreading. Hence, new drugs urgently needed. Phenotypic compound screenings have prevailed as leading method discover candidates against diseases. However, publication of complete genome sequences multiple strains, advances in application CRISPR/Cas9 technology, vivo bioluminescence‐based imaging set stage for advancing target‐based discovery. This review analyses limitations narrow pool available presently treating It describes drug‐based clinical trials highlighting most promising leads. Furthermore, presents a focused discussion on important biological pharmacological challenges that discovery programs must overcome advance candidates. Finally, it examines advantages modern research tools designed identify validate essential genes targets, including genomic editing applications imaging.

Language: Английский

Citations

79
TransLeish: Identification of membrane transporters essential for survival of intracellular Leishmania parasites in a systematic gene deletion screen DOI Creative Commons
Andreia Albuquerque-Wendt,

Ciaran J. McCoy,

Rachel Neish

et al.

Nature Communications, Journal Year: 2025, Volume and Issue: 16(1)

Published: Jan. 2, 2025

Abstract For the protozoan parasite Leishmania , completion of its life cycle requires sequential adaptation cellular physiology and nutrient scavenging mechanisms to different environments a sand fly alimentary tract acidic mammalian host cell phagolysosome. Transmembrane transporters are gatekeepers intracellular environments, controlling flux solutes ions across membranes. To discover which vital for survival as amastigote forms, we carried out systematic loss-of-function screen L. mexicana transportome. A total 312 protein components small molecule carriers, ion channels pumps were identified targeted in CRISPR-Cas9 gene deletion promastigote form, yielding 188 viable null mutants. Forty transporter deletions caused significant loss fitness macrophage mouse infections. striking example is Vacuolar H + ATPase (V-ATPase), which, unexpectedly, was dispensable growth vitro but essential disease-causing amastigotes.

Language: Английский

Citations

1
A limitation lifted: A conditional knockdown system reveals essential roles for Polo-like kinase and Aurora kinase 1 in Trypanosoma cruzi cell division DOI Creative Commons
Justin Wiedeman, Ruby E. Harrison, Ronald Drew Etheridge

et al.

Proceedings of the National Academy of Sciences, Journal Year: 2025, Volume and Issue: 122(8)

Published: Feb. 18, 2025

While advances in genome editing technologies have simplified gene disruption many organisms, the study of essential genes requires development conditional or knockdown systems that are not available most organisms. Such is case for Trypanosoma cruzi, a parasite causes Chagas disease, severely neglected tropical disease endemic to Latin America often fatal. Our knowledge identity and their functions T. cruzi has been constrained by historical challenges very basic genetic manipulation absence RNA interference machinery. Here, we describe use self-cleaving sequences conditionally regulate expression cruzi. Using these tools, identified roles Polo-like Aurora kinases cell division, mirroring brucei. Importantly, demonstrate intracellular amastigotes, disease-causing stage its human host. This system enables efficient scalable functional characterization provides framework other nonmodel

Language: Английский

Citations

1
LeishGEdit: A Method for Rapid Gene Knockout and Tagging Using CRISPR-Cas9 DOI
Tom Beneke, Eva Gluenz

Methods in molecular biology, Journal Year: 2019, Volume and Issue: unknown, P. 189 - 210

Published: Jan. 1, 2019

Language: Английский

Citations

64
Essential roles for deubiquitination in Leishmania life cycle progression DOI Creative Commons
Andreas Damianou, Rebecca J. Burge, Carolina Moura Costa Catta‐Preta

et al.

PLoS Pathogens, Journal Year: 2020, Volume and Issue: 16(6), P. e1008455 - e1008455

Published: June 16, 2020

The parasitic protozoan Leishmania requires proteasomal, autophagic and lysosomal proteolytic pathways to enact the extensive cellular remodelling that occurs during its life cycle. proteasome is essential for parasite proliferation, yet little known about requirement ubiquitination/deubiquitination processes in growth differentiation. Activity-based protein profiling of L. mexicana C12, C19 C65 deubiquitinating cysteine peptidases (DUBs) revealed DUB activity remains relatively constant differentiation procyclic promastigote amastigote. However, when cycle phenotyping (bar-seq) was performed on a pool including 15 barcoded null mutants created promastigotes using CRISPR-Cas9, significant loss fitness observed intracellular infection. DUBs 4, 7, 13 are required successful transformation from metacyclic amastigote 3, 5, 6, 8, 10, 11 14 normal proliferation mice. 1, 2, 12 16 viability role DUB2 establishing infection demonstrated DiCre inducible gene deletion vitro vivo. found nucleus interacts with nuclear proteins associated transcription/chromatin dynamics, mRNA splicing capping. has broad linkage specificity, cleaving all di-ubiquitin chains except Lys27 Met1. Our study demonstrates crucial play survival amastigotes exquisitely sensitive disruption ubiquitination homeostasis.

Language: Английский

Citations

48
Need for sustainable approaches in antileishmanial drug discovery DOI
Sarah Hendrickx, Guy Caljon, Louis Maes

et al.

Parasitology Research, Journal Year: 2019, Volume and Issue: 118(10), P. 2743 - 2752

Published: Aug. 31, 2019

Language: Английский

Citations

47
Gene editing and scalable functional genomic screening in Leishmania species using the CRISPR/Cas9 cytosine base editor toolbox LeishBASEedit DOI Creative Commons
Markus Engstler, Tom Beneke

eLife, Journal Year: 2023, Volume and Issue: 12

Published: May 24, 2023

CRISPR/Cas9 gene editing has revolutionised loss-of-function experiments in Leishmania , the causative agent of leishmaniasis. As lack a functional non-homologous DNA end joining pathway however, obtaining null mutants typically requires additional donor DNA, selection drug resistance-associated edits or time-consuming isolation clones. Genome-wide screens across different conditions and multiple species are therefore unfeasible at present. Here, we report cytosine base editor (CBE) toolbox that overcomes these limitations. We employed CBEs to introduce STOP codons by converting into thymine created http://www.leishbaseedit.net/ for CBE primer design kinetoplastids. Through reporter assays targeting single- multi-copy genes L. mexicana major donovani infantum demonstrate how this tool can efficiently generate expressing just one single-guide RNA, reaching up 100% rate non-clonal populations. then generated -optimised successfully targeted an essential plasmid library delivered screen . Since our method does not require double-strand breaks, homologous recombination, clones, believe enables first time genetic via delivery libraries.

Language: Английский

Citations

15