Discovery of a new small-molecule inhibitor of p53–MDM2 interaction using a yeast-based approach

Leão, Miguel; Pereira, Clara; Bisio, Alessandra; Ciribilli, Yari; Paiva, Ana Mafalda; Machado, Neuza; Palmeira, Andreia; Fernandes, Miguel X.; Sousa, Emı́lia; Pinto, Madalena; Inga, Alberto; Saraiva, Lucı́lia

doi:10.1016/j.bcp.2013.01.032

Cited by 55 publications

(52 citation statements)

References 36 publications

(66 reference statements)

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“…Saccharomyces cerevisiae (strain CG379) expressing human wt p53 alone and combined with human MDM2 or MDMX were obtained in previous works [6,7]. For expression of human proteins (routinely grown in minimal selective medium), cells were diluted to 0.05 OD 600 in selective induction medium containing 2% (w/w) galactose, 1% (w/w) raffinose, 0.7% (w/w) yeast nitrogen base without amino acids from Difco (Quilaban, Sintra, Portugal) and all the amino acids required for yeast growth (50 g/mL) except leucine and tryptophan.…”

Section: Yeast Targeted Screening Assaymentioning

confidence: 99%

“…Whole protein extracts from yeast and human cell lines were obtained as described [7,8]. For mitochondrial and cytosolic fractions of human tumor cell lines, the Mitochondrial Fractionation Kit from Active Motif (Frilabo, Porto, Portugal) was used according to the manufacturer's instructions.…”

Section: Western Blot Analysismentioning

confidence: 99%

“…Recently, our group developed a new approach, based on yeast cells co-expressing human p53 and MDM2 or MDMX, for the targeted screening of inhibitors of the p53 interaction with MDM2 [6] and MDMX [7]. The efficacy of these cell systems for the screening of p53-MDMs interaction inhibitors was demonstrated by the identification of two new small molecule inhibitors of the p53-MDM2 interaction, a pyranoxanthone [7] and an oxazoloisoindolinone [8].…”

Section: Introductionmentioning

confidence: 98%

“…The efficacy of these cell systems for the screening of p53-MDMs interaction inhibitors was demonstrated by the identification of two new small molecule inhibitors of the p53-MDM2 interaction, a pyranoxanthone [7] and an oxazoloisoindolinone [8].…”

Section: Introductionmentioning

confidence: 99%

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A tryptophanol-derived oxazolopiperidone lactam is cytotoxic against tumors via inhibition of p53 interaction with murine double minute proteins

Soares

Raimundo

Pereira

et al. 2015

Pharmacological Research

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Inactivation of the p53 tumor suppressor protein by interaction with murine double minute (MDM) proteins, MDM2 and MDMX, is a common event in human tumors expressing wild-type p53. In these tumors, the simultaneous inhibition of these interactions with MDMs, for a full p53 reactivation, represents a promising anticancer strategy. Herein, we report the identification of a dual inhibitor of the p53 interaction with MDM2 and MDMX, the (S)-tryptophanol derivative OXAZ-1, from the screening of a small library of enantiopure tryptophanol-derived oxazolopiperidone lactams, using a yeast-based assay. With human colon adenocarcinoma HCT116 cell lines expressing wild-type p53 (HCT116 p53(+/+)) and its p53-null isogenic derivative (HCT116 p53(-/-)), it was shown that OXAZ-1 induced a p53-dependent tumor growth-inhibitory effect. In fact, OXAZ-1 induced p53 stabilization, up-regulated p53 transcription targets, such as MDM2, MDMX, p21, Puma and Bax, and led to PARP cleavage, in p53(+/+), but not in p53(-/-), HCT116 cells. In addition, similar tumor cytotoxic effects were observed for OXAZ-1 against MDMX-overexpressing breast adenocarcinoma MCF-7 tumor cells, commonly described as highly resistant to MDM2-only inhibitors. In HCT116 p53(+/+) cells, the disruption of the p53 interaction with MDMs by OXAZ-1 was further confirmed by co-immunoprecipitation. It was also shown that OXAZ-1 potently triggered a p53-dependent mitochondria-mediated apoptosis, characterized by reactive oxygen species generation, mitochondrial membrane potential dissipation, Bax translocation to mitochondria, and cytochrome c release, and exhibited a p53-dependent synergistic effect with conventional chemotherapeutic drugs. Collectively, in this work, a novel selective activator of the p53 pathway is reported with promising antitumor properties to be explored either alone or combined with conventional chemotherapeutic drugs. Moreover, OXAZ-1 may represent a promising starting scaffold to search for new dual inhibitors of the p53-MDMs interaction.

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Section: Yeast Targeted Screening Assaymentioning

confidence: 99%

Section: Western Blot Analysismentioning

confidence: 99%

Section: Introductionmentioning

confidence: 98%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

A tryptophanol-derived oxazolopiperidone lactam is cytotoxic against tumors via inhibition of p53 interaction with murine double minute proteins

Soares

Raimundo

Pereira

et al. 2015

Pharmacological Research

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“…1), presented significant antiproliferative and apoptotic inducing effects (Paiva et al, 2012;Palmeira et al, 2010) in human tumor cell lines. Both ␣-mangostin (Leão et al, 2013a) and compound 1 (Leão et al, 2013b) were shown to be promising inhibitors of p53-MDM2 interaction, with compound 1 showing the highest inhibitory activity in a yeast target-based assay, mimicking the activity of known p53 activators. In addition, compound 1 was shown to inhibit P-glycoprotein in leukemia cells and presented an apparently high permeability coefficient across the human colon cancer cell line (Caco-2) .…”

Section: Introductionmentioning

confidence: 99%

Development of noncytotoxic PLGA nanoparticles to improve the effect of a new inhibitor of p53–MDM2 interaction

Paiva

Pinto

Teixeira

et al. 2013

International Journal of Pharmaceutics

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a b s t r a c tOne possible approach to overcome solubility complications and enhance the biological activity of drugs is their incorporation into drug delivery systems. Within this scope, several nanosphere and nanocapsule formulations of a new inhibitor of p53-MDM2 interaction (xanthone 1) were developed and their physicochemical properties analyzed. Through the investigation of the effect of several empty nanoparticles on the growth of MCF-7 cells, it was possible to observe that four out of five formulations were cytotoxic and that some correlations between the toxic potential of these polymeric nanoparticles and their properties/composition could be extrapolated. One empty formulation of nanocapsules developed by emulsification/solvent evaporation and containing PLGA, PVA and Mygliol ® 812 was found to be noncytotoxic to this cell line. The corresponding compound 1-loaded nanocapsules showed an incorporation efficiency of 77% and revealed to be more potent than the free drug against cell growth inhibition, which may be related to the enhancement in its intracellular delivery. In an integrative study, the intracellular uptake of nanocapsules was confirmed using fluorescent 6-coumarin and well as compound 1 release from nanocapsules. Overall, it was possible to enhance the effect of the hit inhibitor of p53-MDM2 interaction through the development of suitable noncytotoxic polymeric nanoparticles.

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Medicinal Chemistry Strategies to Disrupt the p53–MDM2/MDMX Interaction

Lemos

Leão

Soares

et al. 2016

Medicinal Research Reviews

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The growth inhibitory activity of p53 tumor suppressor is tightly regulated by interaction with two negative regulatory proteins, murine double minute 2 (MDM2) and X (MDMX), which are overexpressed in about half of all human tumors. The elucidation of crystallographic structures of MDM2/MDMX complexes with p53 has been pivotal for the identification of several classes of inhibitors of the p53-MDM2/MDMX interaction. The present review provides in silico strategies and screening approaches used in drug discovery as well as an overview of the most relevant classes of small-molecule inhibitors of the p53-MDM2/MDMX interaction, their progress in pipeline, and highlights particularities of each class of inhibitors. Most of the progress made with high-throughput screening has led to the development of inhibitors belonging to the cis-imidazoline, piperidinone, and spiro-oxindole series. However, novel potent and selective classes of inhibitors of the p53-MDM2 interaction with promising antitumor activity are emerging. Even with the discovery of the 3D structure of complex p53-MDMX, only two small molecules were reported as selective p53-MDMX antagonists, WK298 and SJ-172550. Dual inhibition of the p53-MDM2/MDMX interaction has shown to be an alternative approach since it results in full activation of the p53-dependent pathway. The knowledge of structural requirements crucial to the development of small-molecule inhibitors of the p53-MDMs interactions has enabled the identification of novel antitumor agents with improved in vivo efficacy.

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Discovery of a new small-molecule inhibitor of p53–MDM2 interaction using a yeast-based approach

Cited by 55 publications

References 36 publications

A tryptophanol-derived oxazolopiperidone lactam is cytotoxic against tumors via inhibition of p53 interaction with murine double minute proteins

A tryptophanol-derived oxazolopiperidone lactam is cytotoxic against tumors via inhibition of p53 interaction with murine double minute proteins

Development of noncytotoxic PLGA nanoparticles to improve the effect of a new inhibitor of p53–MDM2 interaction

Medicinal Chemistry Strategies to Disrupt the p53–MDM2/MDMX Interaction

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