Search of Allosteric Inhibitors and Associated Proteins of an AKT-like Kinase from Trypanosoma cruzi

Ochoa, Rodrigo; Rocha-Roa, Cristian; Marín-Villa, Marcel; Robledo, Sara M.; Varela-M, Rubén E.

doi:10.3390/ijms19123951

Cited by 7 publications

(13 citation statements)

References 60 publications

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“…Making use of the approaches above, molecular and cellular studies of Leishmania spp. and T. cruzi AKT-like serine/threonine kinase performed in our laboratory have led to results supporting a potential role of this enzyme as a drug target in the treatment of leishmaniasis and Chagas disease [28][29][30][31]. AKT, also known as PKB, is a pivotal molecule that mediates the PI3K/AKT/mTOR signaling pathway, known to play a key role in signal transduction pathways activated in response to growth factors or insulin, thereby contributing to different cell functions such as nutrient metabolism, cell growth, and apoptosis in humans [32].…”

Section: Introductionmentioning

confidence: 78%

In Silico, In Vitro, and Pharmacokinetic Studies of UBMC-4, a Potential Novel Compound for Treating against Trypanosoma cruzi

et al. 2022

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The lack of therapeutic alternatives for the treatment of Chagas disease, a neglected disease, drives the discovery of new drugs with trypanocidal activity. Consequently, we conducted in vitro studies using UBMC-4, a potential Trypanosoma cruzi AKT-like pleckstrin homology (PH) domain inhibitory compound found using bioinformatics tools. The half effective concentration (EC50) on intracellular amastigotes was determined at 1.85 ± 1 μM showing low cytotoxicity (LC50) > 40 μM on human cell lines tested. In order to study the lethal effect caused by the compound on epimastigotes, morphological changes were assessed by scanning and transmission electron microscopy. Progressive alterations such as flagellum inactivation, cell size reduction, nuclear structure alteration, condensation of chromatin towards the nuclear periphery, vacuole formation, and mitochondrial swelling with kinetoplast integrity loss were evidenced. In addition, apoptosis-like markers in T. cruzi were assessed by flow cytometry, demonstrating that the effect of UBMC-4 on T. cruzi AKT-like kinase reduced the tolerance to nutritional stress-triggered, apoptosis-like events, including DNA fragmentation, mitochondrial damage, and loss of plasma membrane integrity. After this, UBMC-4 was formulated for oral administration and pharmacokinetics were analyzed in a mouse model. Finally, upon oral administration of 200 mg/kg in mice, we found that a UBMC-4 plasma concentration remaining in circulation beyond 24 h after administration is well described by the two-compartment model. We conclude that UBMC-4 has an effective trypanocidal activity in vitro at low concentrations and this effect is evident in T. cruzi cell structures. In mice, UBMC-4 was well absorbed and reached plasma concentrations higher than the EC50, showing features that would aid in developing a new drug to treat Chagas disease.

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Section: Introductionmentioning

confidence: 78%

In Silico, In Vitro, and Pharmacokinetic Studies of UBMC-4, a Potential Novel Compound for Treating against Trypanosoma cruzi

et al. 2022

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“…In the case of trypanosomatids, cytoplasmic overexpression of SIR2 prevents programmed cell death. Also, the anti-trypanosomatid activity of certain Akt-like protein inhibitors suggests the anti-apoptotic and cell survival role of the TriTryp PI3K/AKT pathway, which has GSK-3 as a substrate ( Varela-M et al, 2017 ; Tirado-Duarte et al., 2018 ; Ochoa et al., 2018 ). The anti-trypanosomatid activity of GSK3 inhibitors proposes selective inhibition of the GSK-3 pathway, which is necessary for cell survival ( Ojo et al., 2008 ; Martínez de Iturrate et al., 2020 ).…”

Section: Discussionmentioning

confidence: 99%

The ultimate fate determinants of drug induced cell-death mechanisms in Trypanosomatids

Das

Saha

BoseDasgupta

2021

International Journal for Parasitology: Drugs and Drug Resistan

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Chemotherapy constitutes a major part of modern-day therapy for infectious and chronic diseases. A drug is said to be effective if it can inhibit its target, induce stress, and thereby trigger an array of cell death pathways in the form of programmed cell death, autophagy, necrosis, etc. Chemotherapy is the only treatment choice against trypanosomatid diseases like Leishmaniasis, Chagas disease, and sleeping sickness. Anti-trypanosomatid drugs can induce various cell death phenotypes depending upon the drug dose and growth stage of the parasites. The mechanisms and pathways triggering cell death in Trypanosomatids serve to help identify potential targets for the development of effective anti-trypanosomatids. Studies show that the key proteins involved in cell death of trypanosomatids are metacaspases, Endonuclease G, Apoptosis-Inducing Factor, cysteine proteases, serine proteases, antioxidant systems, etc. Unlike higher eukaryotes, these organisms either lack the complete set of effectors involved in cell death pathways, or are yet to be deciphered. A detailed summary of the existing knowledge of different drug-induced cell death pathways would help identify the lacuna in each of these pathways and therefore open new avenues for research and thereby new therapeutic targets to explore. The cell death pathway associated complexities in metazoans are absent in trypanosomatids; hence this summary can also help understand the trigger points as well as cross-talk between these pathways. Here we provide an in-depth overview of the existing knowledge of these drug-induced trypanosomatid cell death pathways, describe their associated physiological changes, and suggest potential interconnections amongst them.

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“…Survival to these changes is essential for cell cycle and disease progression in human and nonhuman reservoirs. Specific drug inhibitors of either the mammalian PI3K/AKT pathway (i.e., perifosine or NVPBEZ235) or the Leishmania AKT (UBMC1-4) [23,24] have been proven to be effective against trypanosomatids, inducing apoptosis cell death and negatively affecting intracellular proliferation of amastigotes [13].…”

Section: Literature Search Of the Selected Kinasesmentioning

confidence: 99%

“…Moreover, hybrid methodologies are available to include the protein flexibility in an indirect way using a molecular dynamics simulation of the target previous to the screening [21]. These methods have been applied in previous projects using the model of an AKT-like kinase of Leishmania and Trypanosoma [22,23], where a list of molecules were evaluated experimentally with promising results [24].…”

Section: Introductionmentioning

confidence: 99%

Identification of Potential Kinase Inhibitors within the PI3K/AKT Pathway of Leishmania Species

et al. 2021

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Leishmaniasis is a public health disease that requires the development of more effective treatments and the identification of novel molecular targets. Since blocking the PI3K/AKT pathway has been successfully studied as an effective anticancer strategy for decades, we examined whether the same approach would also be feasible in Leishmania due to their high amount and diverse set of annotated proteins. Here, we used a best reciprocal hits protocol to identify potential protein kinase homologues in an annotated human PI3K/AKT pathway. We calculated their ligandibility based on available bioactivity data of the reported homologues and modelled their 3D structures to estimate the druggability of their binding pockets. The models were used to run a virtual screening method with molecular docking. We found and studied five protein kinases in five different Leishmania species, which are AKT, CDK, AMPK, mTOR and GSK3 homologues from the studied pathways. The compounds found for different enzymes and species were analysed and suggested as starting point scaffolds for the design of inhibitors. We studied the kinases’ participation in protein–protein interaction networks, and the potential deleterious effects, if inhibited, were supported with the literature. In the case of Leishmania GSK3, an inhibitor of its human counterpart, prioritized by our method, was validated in vitro to test its anti-Leishmania activity and indirectly infer the presence of the enzyme in the parasite. The analysis contributes to improving the knowledge about the presence of similar signalling pathways in Leishmania, as well as the discovery of compounds acting against any of these kinases as potential molecular targets in the parasite.

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Search of Allosteric Inhibitors and Associated Proteins of an AKT-like Kinase from Trypanosoma cruzi

Cited by 7 publications

References 60 publications

In Silico, In Vitro, and Pharmacokinetic Studies of UBMC-4, a Potential Novel Compound for Treating against Trypanosoma cruzi

In Silico, In Vitro, and Pharmacokinetic Studies of UBMC-4, a Potential Novel Compound for Treating against Trypanosoma cruzi

The ultimate fate determinants of drug induced cell-death mechanisms in Trypanosomatids

Identification of Potential Kinase Inhibitors within the PI3K/AKT Pathway of Leishmania Species

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