A non-commercial approach for the generation of transgenicLeishmania tarentolaeand its application in antileishmanial drug discovery

SUMMARYLeishmaniasis is a parasitic infection caused by several species of the genusLeishmaniathat is considered as a neglected disease. Drug development process requires a robust and updated high-throughput technology to the evaluation of candidate compounds that imply the manipulation of the pathogenic species of the parasite in the laboratory. Therefore, it is restricted to trained personal and level II biosafety environments. However, it has been established the utility ofLeishmania tarentolaeas a model forin vitroscreening of antileishmanial agents without the necessity of level II biosafety setups. In parallel the transfection ofLeishmaniaparasites with reporter genes as the eGFP using non-commercial integration vectors like the pIRmcs3(−) has proved to be a powerful tool for the implementation of semi automatized high-throughput platforms for the evaluation of antileishmanial compounds. Here we report the generation of a newL. tarentolaestrain overexpressing the eGFP gene harboured by the non-commercial vector pIR3(−). We also demonstrate its utility for the semi-automatized screening of antileshmanial compounds in intracellular forms of theL. tarentolaeparasite.

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Challenges and Tools for In Vitro Leishmania Exploratory Screening in the Drug Development Process: An Updated Review

Pathogens ◽

10.3390/pathogens10121608 ◽

2021 ◽

Vol 10 (12) ◽

pp. 1608

Author(s):

Anita Cohen ◽

Nadine Azas

Keyword(s):

Side Effects ◽

Drug Development ◽

Development Process ◽

Screening Tools ◽

Sand Fly ◽

Drug Development Process ◽

Major Health Problem ◽

Vector Borne Diseases ◽

Antileishmanial Drug

Leishmaniases are a group of vector-borne diseases caused by infection with the protozoan parasites Leishmania spp. Some of them, such as Mediterranean visceral leishmaniasis, are zoonotic diseases transmitted from vertebrate to vertebrate by a hematophagous insect, the sand fly. As there is an endemic in more than 90 countries worldwide, this complex and major health problem has different clinical forms depending on the parasite species involved, with the visceral form being the most worrying since it is fatal when left untreated. Nevertheless, currently available antileishmanial therapies are significantly limited (low efficacy, toxicity, adverse side effects, drug-resistance, length of treatment, and cost), so there is an urgent need to discover new compounds with antileishmanial activity, which are ideally inexpensive and orally administrable with few side effects and a novel mechanism of action. Therefore, various powerful approaches were recently applied in many interesting antileishmanial drug development programs. The objective of this review is to focus on the very first step in developing a potential drug and to identify the exploratory methods currently used to screen in vitro hit compounds and the challenges involved, particularly in terms of harmonizing the results of work carried out by different research teams. This review also aims to identify innovative screening tools and methods for more extensive use in the drug development process.

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Microphysiological systems: What it takes for community adoption

Experimental Biology and Medicine ◽

10.1177/15353702211008872 ◽

2021 ◽

pp. 153537022110088

Author(s):

Passley Hargrove-Grimes ◽

Lucie A Low ◽

Danilo A Tagle

Keyword(s):

Rare Diseases ◽

Development Process ◽

Emerging Technology ◽

National Institutes Of Health ◽

Drug Development Process ◽

Microphysiological Systems ◽

Pediatric Populations ◽

Neural Disorders ◽

Future Path

Microphysiological systems (MPS) are promising in vitro tools which could substantially improve the drug development process, particularly for underserved patient populations such as those with rare diseases, neural disorders, and diseases impacting pediatric populations. Currently, one of the major goals of the National Institutes of Health MPS program, led by the National Center for Advancing Translational Sciences (NCATS), is to demonstrate the utility of this emerging technology and help support the path to community adoption. However, community adoption of MPS technology has been hindered by a variety of factors including biological and technological challenges in device creation, issues with validation and standardization of MPS technology, and potential complications related to commercialization. In this brief Minireview, we offer an NCATS perspective on what current barriers exist to MPS adoption and provide an outlook on the future path to adoption of these in vitro tools.

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The 18th FRAME Annual Lecture, October 2019: Human In Silico Trials in Pharmacology

Alternatives to Laboratory Animals ◽

10.1177/0261192919896356 ◽

2019 ◽

Vol 47 (5-6) ◽

pp. 221-227

Author(s):

Blanca Rodriguez

Keyword(s):

Drug Development ◽

In Silico ◽

Drug Testing ◽

Development Process ◽

Computer Modelling ◽

Current Knowledge ◽

Magnetic Resonance Images ◽

Population Heterogeneity ◽

Drug Development Process

Safety and efficacy testing is a crucial part of the drug development process, and several different methods are used to obtain the necessary data (e.g. in vitro testing, animal trials and clinical trials). Our group has been investigating the potential of modelling and simulation as an alternative approach to some of the methods used for testing drugs for cardiac effects. To achieve our goal of developing and promoting novel approaches in drug development, we formed multidisciplinary collaborations that included clinicians, computer scientists and biologists. Our in silico models are based on human data (e.g. magnetic resonance images, electrocardiogram) and on current knowledge of human electrophysiology, thus generating predictions that are directly applicable to humans. Such models are a particularly powerful tool because they encompass different sources of population heterogeneity, which is crucial for drug testing and for assessing how interindividual variability might affect clinical endpoints. Our group has shown that computer modelling can be used to predict the effects of a test drug in a virtual population or in combination with machine learning to predict different phenotypes when a drug is given to a diseased population. Furthermore, our user-friendly drug testing software is freely available and is being adopted by industry in their drug development process. We have been engaging with industry and regulators to show that our models can contribute to the replacement of animals in drug development. Our ambition is to generate models for simulation of different diseases and therapies for investigations from subcellular to whole organ.

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Synthesis, characterization and toxicity studies of pyridinecarboxaldehydes and L-tryptophan derived Schiff bases and corresponding copper (II) complexes

F1000Research ◽

10.12688/f1000research.9226.1 ◽

2016 ◽

Vol 5 ◽

pp. 1921 ◽

Cited By ~ 4

Author(s):

Margarita Malakyan ◽

Nelly Babayan ◽

Ruzanna Grigoryan ◽

Natalya Sarkisyan ◽

Vahan Tonoyan ◽

...

Keyword(s):

Schiff Base ◽

Drug Development ◽

Schiff Bases ◽

Copper Complexes ◽

Development Process ◽

Biological Activities ◽

Drug Development Process ◽

Relationship Analysis

Schiff bases and their metal-complexes are versatile compounds exhibiting a broad range of biological activities and thus actively used in the drug development process. The aim of the present study was the synthesis and characterization of new Schiff bases and their copper (II) complexes, derived from L-tryptophan and isomeric (2-; 3-; 4-) pyridinecarboxaldehydes, as well as the assessment of their toxicity in vitro. The optimal conditions of the Schiff base synthesis resulting in up to 75-85% yield of target products were identified. The structure-activity relationship analysis indicated that the location of the carboxaldehyde group at 2-, 3- or 4-position with regard to nitrogen of the pyridine ring in aldehyde component of the L-tryptophan derivative Schiff bases and corresponding copper complexes essentially change the biological activity of the compounds. The carboxaldehyde group at 2- and 4-positions leads to the higher cytotoxic activity, than that of at 3-position, and the presence of the copper in the complexes increases the cytotoxicity. Based on toxicity classification data, the compounds with non-toxic profile were identified, which can be used as new entities in the drug development process using Schiff base scaffold.

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Directed In Vitro Evolution of Reporter Genes Based on Semi-Rational Design and High-Throughput Screening

Methods in Molecular Biology - In Vitro Mutagenesis Protocols ◽

10.1007/978-1-60761-652-8_18 ◽

2010 ◽

pp. 239-256 ◽

Cited By ~ 4

Author(s):

Ai-Sheng Xiong ◽

Quan-Hong Yao ◽

Ri-He Peng ◽

Zong-Ming Cheng

Keyword(s):

High Throughput ◽

High Throughput Screening ◽

Rational Design ◽

Reporter Genes ◽

In Vitro Evolution

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In VitroScreening Test Using Leishmania Promastigotes Stably Expressing mCherry Protein

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.02224-13 ◽

2014 ◽

Vol 58 (3) ◽

pp. 1825-1828 ◽

Cited By ~ 11

Author(s):

Paola Vacchina ◽

Miguel A. Morales

Keyword(s):

High Throughput ◽

Drug Screening ◽

Leishmania Donovani ◽

Leishmania Major ◽

Content Type ◽

Minimal Sample ◽

Antileishmanial Drug ◽

Leishmania Promastigotes

ABSTRACTTransgenicLeishmania majorandLeishmania donovaniaxenic promastigotes constitutively expressing mCherry were used forin vitroantileishmanial drug screening. This method requires minimal sample manipulation and can be easily adapted to automatic drug tests, allowing primary high-throughput screenings without the need for expensive and sophisticated instruments.

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