2,6,9-Trisubstituted purines as CRK3 kinase inhibitors with antileishmanial activity in vitro

Řezníčková, Eva; Popa, Alexandr; Gucký, Tomáš; Zatloukal, Marek; Havlı́cěk, Libor; Bazgier, Václav; Berka, Karel; Jorda, Radek; Popa, Igor; Nasereddin, Abedelmajeed; Jaffe, Charles L.; Kryštof, Vladimı́r; Strnad, Miroslav

doi:10.1016/j.bmcl.2015.04.030

Cited by 11 publications

(5 citation statements)

References 20 publications

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“…Previous attempts to impair CRK3 function in Leishmania by treatment with protein kinase inhibitors may have resulted in off‐target effects (Cleghorn et al ., ; Efstathiou et al ., ; Grant et al ., ; Jorda et al ., ; Reichwald et al ., ; Řezníčková et al ., ). Here the utilization of diCre mediated gene deletion enabled the effect of CRK3 depletion on the cell cycle to be investigated.…”

Section: Resultsmentioning

confidence: 97%

“…In particular, the CDK‐related kinase CRK3 has been demonstrated as being important for regulation of the L. mexicana promastigote cell cycle by existing genetic manipulation techniques and cell cycle arrest following treatment with CDK inhibitors (Hassan et al ., ; Grant et al ., 1998; 2004). Recombinant protein kinase activity assays (Gomes et al ., ) and yeast recovery mutants (Wang et al ., ) have provided further validation of CRK3 as a drug target, leading to the identification and synthesis of a number of CRK3 inhibitors (Cleghorn et al ., ; Goyal et al ., ; Grant et al ., ; Řezníčková et al ., ; Walker et al ., ). Regulation of CRK3 expression in L. mexicana is desirable to further assess its function in both procyclic promastigote and amastigote life cycle stages, however, no system exists for conditional deletion of essential genes.…”

Section: Introductionmentioning

confidence: 99%

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Conditional gene deletion with DiCre demonstrates an essential role for CRK3 in Leishmania mexicana cell cycle regulation

Duncan

Myburgh

Philipon

et al. 2016

Molecular Microbiology

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Summary Leishmania mexicana has a large family of cyclin‐dependent kinases (CDKs) that reflect the complex interplay between cell cycle and life cycle progression. Evidence from previous studies indicated that Cdc2‐related kinase 3 (CRK3) in complex with the cyclin CYC6 is a functional homologue of the major cell cycle regulator CDK1, yet definitive genetic evidence for an essential role in parasite proliferation is lacking. To address this, we have implemented an inducible gene deletion system based on a dimerised Cre recombinase (diCre) to target CRK3 and elucidate its role in the cell cycle of L. mexicana. Induction of diCre activity in promastigotes with rapamycin resulted in efficient deletion of floxed CRK3, resulting in G2/M growth arrest. Co‐expression of a CRK3 transgene during rapamycin‐induced deletion of CRK3 resulted in complementation of growth, whereas expression of an active site CRK3 T178E mutant did not, showing that protein kinase activity is crucial for CRK3 function. Inducible deletion of CRK3 in stationary phase promastigotes resulted in attenuated growth in mice, thereby confirming CRK3 as a useful therapeutic target and diCre as a valuable new tool for analyzing essential genes in Leishmania.

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Section: Resultsmentioning

confidence: 97%

Section: Introductionmentioning

confidence: 99%

Conditional gene deletion with DiCre demonstrates an essential role for CRK3 in Leishmania mexicana cell cycle regulation

Duncan

Myburgh

Philipon

et al. 2016

Molecular Microbiology

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“…The cyclin-dependent kinases 2 – CRK1 and CRK3 were experimentally validated in L. mexicana as essential proteins [59] with fundamental roles during cell cycle progression [60]. Also, many inhibitor-driven studies have been carried in order to reach CRK3 inhibition, but few displayed in vitro activity [[61], [62], [63], [64]]. Recombinant GSK3 was expressed and purified in L. major and L. infantum.…”

Section: Discussionmentioning

confidence: 99%

Unveiling the Kinomes of Leishmania infantum and L. braziliensis Empowers the Discovery of New Kinase Targets and Antileishmanial Compounds

Borba

Silva

Ramos

et al. 2019

Computational and Structural Biotechnology Journal

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Leishmaniasis is a neglected tropical disease caused by parasites of the genus Leishmania (NTD) endemic in 98 countries. Although some drugs are available, current treatments deal with issues such as toxicity, low efficacy, and emergence of resistance. Therefore, there is an urgent need to identify new targets for the development of new antileishmanial drugs. Protein kinases (PKs), which play an essential role in many biological processes, have become potential drug targets for many parasitic diseases. A refined bioinformatics pipeline was applied in order to define and compare the kinomes of L. infantum and L. braziliensis, species that cause cutaneous and visceral manifestations of leishmaniasis in the Americas, the latter being potentially fatal if untreated. Respectively, 224 and 221 PKs were identified in L. infantum and L. braziliensis overall. Almost all unclassified eukaryotic PKs were assigned to six of nine major kinase groups and, consequently, most have been classified into family and subfamily. Furthermore, revealing the kinomes for both Leishmania species allowed for the prioritization of potential drug targets that could be explored for discovering new drugs against leishmaniasis. Finally, we used a drug repurposing approach and prioritized seven approved drugs and investigational compounds to be experimentally tested against Leishmania. Trametinib and NMS-1286937 inhibited the growth of L. infantum and L. braziliensis promastigotes and amastigotes and therefore might be good candidates for the drug repurposing pipeline.

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“…According to the aforementioned, in the last decades, several synthetic procedures to obtain di‐ or trisubstituted purines have been reported, where the positions C‐2, C‐6, and N‐9 are the most explored. The influences of different substituents in these positions in several bioactive purine derivatives have been extensively studied, and according to these results, optimal substituents have been identified [6,7,14–16]. An alkylated side chain at C‐2 position certainly opens the path to a significant range of possible modifications of the parent purines.…”

Section: Introductionmentioning

confidence: 99%

Convergent synthesis, drug target prediction, and docking studies of new 2,6,9‐trisubstituted purine derivatives

Villegas

Satheeshkumar

Ballesteros-Casallas

et al. 2021

Journal of Heterocyclic Chem

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A set of new 2,6,9‐trisubstituted purine derivatives were designed and synthesized from 6‐chloro‐2‐fluoro‐9‐alkyl‐9H‐purine as the key starting material. These purines were synthesized via a multistep protocol and finally subjected to SNAr with various aminopiperidinyl salts. The structures of these compounds were established by substantiating them through spectral techniques like FT‐IR, 1H‐NMR, 13C‐NMR, and 19F‐NMR. In addition, for two purine derivatives, a reverse screening strategy based on ligand similarity (PharmMapper web server) resulted in a set of predicted protein target candidates. Interestingly, for both purines, the main potential target candidates belonged to key proteins involved within signaling pathways that are related to proliferation or survival of cancer cells. These proteins correspond mainly to enzymes and specifically kinases. To check if our purines could be ligands for two kinases involved in cancer, docking studies were performed. The results indicated that these purines fitted in the same cavity of crystalized inhibitors. Therefore, in this work we are developed new purine derivatives that could be good inhibitors of some kinases.

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2,6,9-Trisubstituted purines as CRK3 kinase inhibitors with antileishmanial activity in vitro

Cited by 11 publications

References 20 publications

Conditional gene deletion with DiCre demonstrates an essential role for CRK3 in Leishmania mexicana cell cycle regulation

Conditional gene deletion with DiCre demonstrates an essential role for CRK3 in Leishmania mexicana cell cycle regulation

Unveiling the Kinomes of Leishmania infantum and L. braziliensis Empowers the Discovery of New Kinase Targets and Antileishmanial Compounds

Convergent synthesis, drug target prediction, and docking studies of new 2,6,9‐trisubstituted purine derivatives

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